Current and future therapies for hepatitis C virus infection: from viral proteins to host targets.
Hepatitis C virus (HCV) infection is the most important problem across the world. It causes acute and chronic liver infection. Different approaches are in use to inhibit HCV infection, including small organic compounds, siRNA, shRNA and peptide inhibitors. This review article summarizes the current and future therapies for HCV infection. PubMed and Google Scholar were searched for articles published in English to give an insight into the current inhibitors against this life-threatening virus. HCV NS3/4A protease inhibitors and nucleoside/nucleotide inhibitors of NS5B polymerase are presently in the most progressive stage of clinical development, but they are linked with the development of resistance and viral breakthrough. Boceprevir and telaprevir are the two most important protease inhibitors that have been approved recently for the treatment of HCV infection. These two drugs are now the part of standard-of-care treatment (SOC). There are also many other drugs in phase III of clinical development. When exploring the various host-cell-targeting compounds, the most hopeful results have been demonstrated by cyclophilin inhibitors. The current SOC treatment of HCV infection is Peg-interferon, ribavirin and protease inhibitors (boceprevir or telaprevir). The future treatment of this life-threatening disease must involve combinations of therapies hitting multiple targets of HCV and host factors. It is strongly expected that the near future, treatment of HCV infection will be a combination of direct-acting agents (DAA) without the involvement of interferon to eliminate its side effects.


Distinct neural codes in primate hippocampus and lateral prefrontal cortex during associative learning in virtual environments.
The hippocampus (HPC) and the lateral prefrontal cortex (LPFC) are two cortical areas of the primate brain deemed essential to cognition. Here, we hypothesized that the codes mediating neuronal communication in the HPC and LPFC microcircuits have distinctively evolved to serve plasticity and memory function at different spatiotemporal scales. We used a virtual reality task in which animals selected one of the two targets in the arms of the maze, according to a learned context-color rule. Our results show that during associative learning, HPC principal cells concentrate spikes in bursts, enabling temporal summation and fast synaptic plasticity in small populations of neurons and ultimately facilitating rapid encoding of associative memories. On the other hand, layer II/III LPFC pyramidal cells fire spikes more sparsely distributed over time. The latter would facilitate broadcasting of signals loaded in short-term memory across neuronal populations without necessarily triggering fast synaptic plasticity.


Invasive breast carcinomas of no special type with osteoclast-like giant cells frequently have a luminal phenotype.
Invasive carcinoma of no special type (NST) with osteoclast-like giant cells (OGCs) represents a unique type of breast neoplasm, characterized by the presence of multinucleated OGCs and a vascularized, hemorrhagic stroma. Because of its rarity, the literature regarding this tumor remains limited and a detailed immunophenotype of this tumor has not been established as yet. We report a clinicopathological and immunohistochemical study of 42 patients with invasive carcinoma NST with OGCs. Macroscopically, these tumors presented as a well-delimited red-brown mass. A remarkable feature of the tumor was the presence of OGCs in the fibroblastic or hemorrhagic vascular stroma, as well as in the adjacent tumor nests or glandular lumina. The number of OGCs varied from 8 to 105 per 10 high-power fields with an average of 48. The tumors were well to moderately differentiated. Cribriform architecture was observed in 27 tumors (63 %). All of the 36 available tumors were of luminal phenotype, according to the Ki67 labeling index 89 % luminal A and 11 % luminal B. With a mean follow-up time of 46.4 months, lung metastasis was found in 2 patients (5 %) at 7 and 11 years after the operation, respectively. None of the other cases had presented with evidence of recurrence or metastasis. To the best of our knowledge, this is the largest reported series of invasive carcinoma NST with OGCs as yet. Our study revealed that invasive carcinoma NST with OGCs exhibit a luminal phenotype with luminal A subtype as the major group.


Value, confidence, deliberation: a functional partition of the medial prefrontal cortex demonstrated across rating and choice tasks.
Deciding about courses of action involves minimizing costs and maximizing benefits. Decision neuroscience studies have implicated both the ventral and dorsal medial prefrontal cortex (vmPFC and dmPFC) in signaling goal value and action cost, but the precise functional role of these regions is still a matter of debate. Here, we suggest a more general functional partition that applies not only to decisions but also to judgments about goal value (expected reward) and action cost (expected effort). In this conceptual framework, cognitive representations related to options (reward value and effort cost) are dissociated from metacognitive representations (confidence and deliberation) related to solving the task (providing a judgment or making a choice). We used an original approach aiming at identifying consistencies across several preference tasks, from likeability ratings to binary decisions involving both attribute integration and option comparison. fMRI results in human male and female participants confirmed the vmPFC as a generic valuation system, its activity increasing with reward value and decreasing with effort cost. In contrast, more dorsal regions were not concerned with the valuation of options but with metacognitive variables, confidence being reflected in mPFC activity and deliberation time in dmPFC activity. Thus, there was a dissociation between the effort attached to choice options (represented in the vmPFC) and the effort invested in deliberation (represented in the dmPFC), the latter being expressed in pupil dilation. More generally, assessing commonalities across preference tasks might help reaching a unified view of the neural mechanisms underlying the cost/benefit tradeoffs that drive human behavior.Significance statementDecision neuroscience studies have implicated the medial prefrontal cortex (mPFC) in forming the cognitive representations that drive human choice behavior. However, different studies using different tasks have suggested somewhat inconsistent links between precise computational variables and specific brain regions. Here, we use fMRI to demonstrate a robust functional partition of the mPFC that generalizes across tasks involving an estimation of goal value and/or action cost to provide a judgement or make a choice. This general functional partition makes a critical dissociation between neural representations of decisional factors (the expected costs and benefits attached to a given option) and metacognitive estimates (confidence in the judgment or choice, and effort invested in the deliberation process).


Epidermal growth factor receptor expression in mice skin upon ultraviolet B exposure - Seborrheic Keratosis as a coincidental and unique finding.
Ultraviolet B (UVB) is the most damaging component of sunlight. It rapidly activates the epidermal growth factor receptor (EGFR) and generates reactive oxygen species (ROS) in excessive quantities that quickly overwhelm tissue antioxidants. To demonstrate the effects of UVB radiation on EGFR expression in mice skin and to evaluate the role of antioxidants in the exposed group. After obtaining the approval of the ethical committee, forty mice from BALB/c strain were used in this experiment and were allocated into 3 groups; 10 (control group); 15 (exposure group); and 15 (exposed and treated with antioxidants). Antioxidants were administered through subcutaneous injection. Skin biopsies from all groups were stained with EGFR antibodies. Total antioxidant status (TAS) was evaluated in all groups. The data obtained were analyzed using ANOVA, Duncan's test, and Pearson's Correlation. The highest EGFR expression in exposure group was of score 3(+) (53%). The highest EGFR expression in treatment group was score 0 (40%). Apoptotic bodies and dermal mast cells increased in exposure group while decreased in treatment group. The mean values for TAS were measured for each group; control group = 1.2 mmol/l; exposure group = 0.87 mmol/l; treatment group =1.3 mmol/l. UVB led to Seborrheic Keratosis (SK) in mice through enhancement of EGFR expression. Antioxidants effectively reduced UVB-induced SK, reduced epidermal changes, apoptotic bodies, and decreased dermal mast cells. TAS measurement declined in exposure group, while it was within normal range in most treated cases.


Nucleosome linking number change controlled by acetylation of histones H3 and H4.
High levels of acetylation of lysines in the amino-terminal domains of all four core histones, H2A, H2B, H3, and H4, have been shown to reduce the linking number change per nucleosome core particle in reconstituted minichromosomes (Norton, V. G., Imai, B. S., Yau, P., and Bradbury, E. M. (1989) Cell 57, 449-457). Because there is evidence to suggest that the acetylations of H3 and H4 have functions that are distinct from those of H2A and H2B, we have determined the nucleosome core particle linking number change in minichromosomes containing fully acetylated H3 and H4 and very low levels of acetylation in H2A and H2B. This linking number change was -0.81 +/- 0.05, in close agreement with the linking number change for hyperacetylated nucleosome core particles which contain high levels of acetylation in all four core histones (approximately 70% of full acetylation in H3 and H4). Therefore, high levels of acetylation of H3 and H4 alone are responsible for the reduction in the linking number change per nucleosome core particle.


Zinc monotherapy from time of diagnosis for young pediatric patients with presymptomatic Wilson disease.
In 4 young pediatric patients with presymptomatic Wilson disease, we found zinc monotherapy beginning at time of diagnosis to be safe and highly effective for follow-up intervals between 1 and 2 years. Such maintenance therapy with zinc can maintain urinary copper excretion between 1 and 3 μg · kg(-1) · day.


Extravascular injection of sclerotic agents does not affect vessels in the rat: experimental implications for percutaneous sclerotherapy of arteriovenous malformations.
Sclerotherapy is useful for the treatment of arteriovenous vascular malformations. However, intravascular administration of sclerotic agents into small arteriovenous niduses is often difficult. Extravascular administration of sclerotic agents causes reduction of vascular flow on Doppler echo during clinical sclerotherapy. Therefore, we aimed to investigate whether the extravascular injection of sclerotic agents affects tiny vessels. Animal study. The effect of extravascular injection of sclerotic agents on vessels was investigated using rat femoral and superficial inferior epigastric vessels. After surgical exposure of vessels, absolute ethanol, 5% ethanolamine oleate and 3% polidocanol were injected into perivascular surrounding tissues, and their effect on vessels was evaluated after 14 days using histology and coloured silicone rubber injection. The integrity of the vascular lumen, endothelial cells and vascular patency were not affected by injection of sclerotic agents. Attenuation of vascular flow of an arteriovenous shunt after extravascular injection of sclerotic agents is transient and/or trivial and does not cause disruption of vessels. Therefore, sclerotic agents should be delivered to obtain sufficient destruction of arteriovenous malformation lesions and blood flow.


Improving the quality of small animal brain pinhole SPECT imaging by Bayesian reconstruction.
The possibility of using existing hardware makes pinhole single-photon emission computed tomography (SPECT) attractive when pursuing the ultra-high resolution required for small animal brain imaging. Unfortunately, the poor sensitivity and the heavy weight of the collimator hamper the use of pinhole SPECT in animal studies by generating noisy and misaligned projections. To improve the image quality we have developed a new Bayesian reconstruction method, pinhole median root prior (PH-MRP), which prevents the excessive noise accumulation from the projections to the reconstructed image. The PH-MRP algorithm was used to reconstruct data acquired with our small animal rotating device, which was designed to reduce the rotation orbit misalignments. Phantom experiments were performed to test the device and compare the PH-MRP with the conventional Feldkamp-Davis-Kress (FDK) and pinhole ordered subsets maximum likelihood expectation maximisation (PH-OSEM) reconstruction algorithms. The feasibility of the system for small animal brain imaging was studied with Han-Wistar rats injected with (123)I-epidepride and (99m)Tc-hydroxy methylene diphosphonate. Considering all the experiments, no shape distortions due to orbit misalignments were encountered and remarkable improvements in noise characteristics and also in overall image quality were observed when the PH-MRP was applied instead of the FDK or PH-OSEM. In addition, the proposed methods utilise existing hardware and require only a certain amount of construction and programming work, making them easy to implement.


Biodegradable oxygen-producing manganese-chelated metal organic frameworks for tumor-targeted synergistic chemo/photothermal/ photodynamic therapy.
Photodynamic therapy (PDT) is an effective noninvasive therapeutic strategy that can convert oxygen to highly cytotoxic singlet oxygen (1O2) through the co-localization of excitation light and photosensitizers. However, compromised by the hypoxic tumor microenvironment, the therapeutic efficacy of PDT is reduced seriously. Herein, to overcome tumor-associated hypoxia, and further achieve tumor-targeted synergistic chemotherapy/PDT/photothermal therapy (PTT), we have constructed a biodegradable oxygen-producing nanoplatform (named Ini@PM-HP), which was composed of the porous metal-organic framework (PCN-224(Mn)), the poly (ADP-ribose) polymerase (PARP) inhibitor (Iniparib), and the polydopamine-modified hyaluronic acid (HA-PDA). Since HA can specifically bind to the overexpressed HA receptors (cluster determinant 44, CD44) on tumor cell, Ini@PM-HP prefers to accumulate at the tumor site once injected intravenously. Then iniparib can be released in tumor environment (TME), thereby dysfunctioning DNA damage repair and promoting cell apoptosis. At the same time, the chelating of Mn and tetrakis(4-carboxyphenyl) porphyrin (Mn-TCPP) can generate O2 in situ by reacting with endogenous H2O2, relieving the hypoxic TME and achieving enhanced PDT. Moreover, owing to the high photothermal conversion efficiency of PDA, PTT can be driven by the 808 nm laser irradiation. As systematically demonstrated in vitro and in vivo, this nanotherapeutic approach enables the combined therapy with great inhibition on tumor. Overall, the as-prepared nanoplatform provide a promising strategy to overcome tumor-associated hypoxia, and shows great potential for combination tumor therapy. STATEMENT OF SIGNIFICANCE: A delicately designed biodegradable oxygen-producing nanoplatform Ini@PM-HP is constructed to achieve combination therapy of solid tumors. Taking advantage of the active-targeting, PTT, enhanced PDT and PARPi, this nanotherapeutic approach successfully enables the combined chemo/photothermal/photodynamic therapy with great inhibition of solid tumors.


Quantification of bacterial invasion into adherent cells by flow cytometry.
Quantification of invasive, intracellular bacteria is critical in many areas of cellular microbiology and immunology. We describe a novel and fast approach to determine invasion of bacterial pathogens in adherent cell types such as epithelial cells or fibroblasts based on flow cytometry. Using the CEACAM-mediated uptake of Opa-expressing Neisseria gonorrhoeae as a well-characterized model of bacterial invasion, we demonstrate that the flow cytometry-based method yields results comparable to a standard antibiotic protection assay. Furthermore, the quantification of intracellular bacteria by the novel approach is not biased by intracellular killing of the microbes and correctly discriminates between cell-associated extracellular and bona fide intracellular bacteria. As flow cytometry-based quantification is also applicable to other pathogen-host interactions such as the integrin-mediated internalization of Staphylococcus aureus, this approach provides a fast and convenient alternative for the quantification of bacterial uptake and should be particularly useful in elucidating the molecular mechanisms of pathogen-triggered host cell invasion.


Perfluorooctane sulfonate exposure induces preeclampsia-like syndromes by damaging trophoblast mitochondria in pregnant mice.
Preeclampsia, defined as a hypertensive disorder during pregnancy, is a major cause of maternal and fetal mortality. Observational studies have shown that the exposure of per- and polyfluoroalkyl substances, such as perfluorooctane sulfonate (PFOS), is emerging as a significant environmental factor associated with preeclampsia risk. However, epidemiologic evidence is of correlative in nature, and unable to establish a causal relationship. Here, we established an animal model of PFOS-induced preeclampsia to explore the molecular mechanism of PFOS in placental trophoblast. In the mouse model, PFOS exposure by gavage at a dose of 10 mg/kg/d from embryonic day 7.5-16.5 was sufficient to induce preeclampsia-like symptoms such as hypertension, proteinuria, and renal glomerular endotheliosis, accompanied with placental abnormal stromal collagen deposition. In-vitro experiments of JEG-3 cells, PFOS exposure impaired trophoblast motility including the compromised abilities of migration, invasion and vascularization. Mechanistically, these pathological effects on cells resulted from SLC25A5-mediated mitochondrial damages, characterized by excessive ROS generation, decreased ATP production and mitochondrial membrane potential loss, and accompanied by the activation of p38 MAPK and JNK signaling pathways. This pioneering study provided biological plausibility to the causality verified by the animal model and the in vitro experiments, which indicates that PFOS exposure may cause preeclampsia during pregnancy via impairing trophoblast mitochondria.


Fractures of the tibial pilon treated by open reduction and internal fixation (locking compression plate-less invasive stabilising system): Complications and sequelae.
The aim of this study was to evaluate variables that could be related to complications and sequelae in fractures of the tibial pilon treated by open reduction and internal fixation (ORIF) with a locking compression plate-less invasive stabilising system (LCP-LISS). A total of 137 fractures treated by ORIF in a 7-year period were analysed. The mean follow-up was 3.3 years. We analysed the following variables: age, sex, side, type of fracture, energy of the injury, use of provisional external fixation (EF), time until ORIF, stages of treatment (one or two), surgical approach, type of bone fixation, quality of reduction, use of bone graft, hardware removal, associated fractures (fibula and others), functional results (AOFAS scale), early complications (infection, skin necrosis) and late complications (nonunion, early post-traumatic ankle osteoarthritis [AOA]). According to the AOFAS scale, 30.5% of the results were excellent, 46.7% good, 13.1% fair and 9.7% poor. The rate of infection was 8.7%, and the rate of skin necrosis requiring flap coverage was 15.2%. Furthermore, type 43C3 fractures of the AO classification had a higher rate of skin necrosis and flap coverage. The rate of nonunion was 16.3% (22 cases, 4 aseptic, 18 infected), and the use of a medial plate was related to a higher rate of nonunion than the use of a lateral plate. The rate of early post-traumatic AOA was 13.1%, and open fractures were related to a higher prevalence of nonunion and flap coverage. Both infection and a suboptimal anatomic reduction were related to a higher prevalence of fair and poor results. The anteromedial approach was associated with a higher prevalence of skin necrosis and early post-traumatic AOA than the anterolateral approach. Optimal reduction and stable fixation is paramount to diminishing the rate of complications and sequelae after ORIF (LCP-LISS) of these fractures.


Differentiation of isomeric malonylated flavonoid glyconjugates in plant extracts with UPLC-ESI/MS/MS.
Glycosylation at different hydroxyl groups of flavonoids and acylation of sugar moieties are ubiquitous modifications observed in plants. These modifications give rise to simultaneous presence of numerous isomeric and isobaric compounds in tissues and extracts thereof. To develop UPLC-MS method capable for resolution of isomeric malonylated glycoconjugates of flavonoids and recognition of structural differences. Flavonoid glycoconjugates were extracted from leaves of blue lupin (Lupinus angustifolius L.) plants with 80% methanol. Extracts were analysed using ultraperformance liquid chromatography (UPLC) combined with tandem (quadrupole-time of flight, QToF) mass spectrometry. Differentiation of malonylated glycosides of isoflavones and flavones is demonstrated in this paper. The use of UPLC-MS/MS enabled 38 flavonoid conjugates to be distinguished, including the discrimination of five different isomers of a single 3'-O-methylluteolin glycoside. Additionally, pseudo MS(3) experiments (CID spectra registered at high cone voltages) enabled confirmation of the aglycone structures by comparison of their spectra with those obtained from aglycone standards. Application of UPLC-MS/MS allows separation and identification numerous positional isomers of malonylated glycosides of flavonoids and isoflavonoids in plant material. Provided there is strict control of the MS ionisation parameters, this method may be useful for preparation of a flavonoids spectra database, enabling the inter-laboratory comparison of analytical results.


Identification and validation of biomarkers associated with acute and chronic graft versus host disease.
Graft versus host disease (GVHD) is a major complication of haematopoietic SCT (HSCT). A number of inflammatory cytokines/chemokines are implicated in GVHD and have been identified in numerous single centre studies as potential biomarkers for acute and/or chronic GVHD. In this study, we analysed candidate inflammatory biomarkers (B-cell activating factor (BAFF), interleukin 33 (IL-33), CXCL10 and CXCL11) in a two-centre study. Biomarkers were evaluated pre-transplant and at serial time points post transplant in acute and chronic GVHD patient sera with time-matched control samples from patients without GVHD. Further validation was performed using the human skin explant assay, clinical GVHD biopsies and mRNA expression analysis. BAFF was significantly increased pre-transplant. BAFF, IL-33, CXCL10 and CXCL11 showed increased levels in acute GVHD patient sera and high protein expression in grades II-III of the in vitro skin explant graft versus host reaction (GVHR) group. BAFF, CXCL10 and CXCL11 also showed increased mRNA expression levels in clinical biopsies compared with the no/low-grade GVHD group. BAFF, CXCL10 and CXCL11 levels were increased in chronic GVHD patient sera. The results identify BAFF and CXCL10 as predictive biomarkers for acute GVHD and BAFF, CXCL10 and CXCL11 as useful diagnostic biomarkers for acute GVHD and chronic GVHD.


Mucosal permeability and mast cells as targets for functional gastrointestinal disorders.
The intestinal mucosa is constantly exposed to harmful luminal content, and uptake is closely controlled and regulated by neuro-immune factors. If control is broken, it might lead to ongoing enhanced mucosal permeability, potentially resulting in functional gastrointestinal disorders. The importance of mast cells in the regulation of the mucosal barrier has become obvious, and increased numbers and more activated mast cells have been observed in irritable bowel syndrome, functional dyspepsia and gastroesophageal reflux disease. To target the disturbed mucosal permeability, directly or via mast cells, is therefore currently of major interest. For example, administration of mast cell stabilizers and probiotics have shown promising effects in patients with functional gastrointestinal disorders.


SSeCKS/Gravin/AKAP12 Inhibits PKCζ-Mediated Reduction of ERK5 Transactivation to Prevent Endotoxin-Induced Vascular dysfunction.
SSeCKS/Gravin/AKAP12 is a protein kinase C (PKC) substrate that inhibits the activity of PKC through binding with it. SSeCKS is expressed in vascular endothelial cells (ECs). The atypical PKC isoform ζ (PKCζ) is a pathologic mediator of endothelial dysfunction. However, the functional significance of SSeCKS/PKCζ dimerization in the vascular endothelium remains poorly understood. Given this background, we investigated the effects of SSeCKS on endothelial dysfunction and elucidated the possible mechanism involved. Vascular endothelial dysfunction and inflammatory changes were induced by treatment with bacterial endotoxin lipopolysaccharide (LPS, a vascular endothelial toxicity inducer). LPS can increase the level of SSeCKS. However, we also found that depletion of SSeCKS aggravated the LPS-induced vascular endothelial dysfunction, upregulated pro-inflammatory proteins and phosphorylation level of PKCζ, increased ROS formation, decreased extracellular-signal-regulated kinase 5 (ERK5) transcriptional activity, and reduced eNOS expression. Further examination revealed that depletion of SSeCKS increased PKCζ/ERK5 dimerization. These findings provide preliminary evidence that the expression of SSeCKS induced by LPS, as a negative feedback mechanism, has the potential to improve endothelium-dependent relaxation in vascular disease conditions by inhibiting PKCζ-mediated reduction of ERK5 transactivation.


Distinct roles of neuroligin-1 and SynCAM1 in synapse formation and function in primary hippocampal neuronal cultures.
Neuroligins are a family of cell adhesion molecules critical in establishing proper central nervous system connectivity; disruption of neuroligin signaling in vivo precipitates a broad range of cognitive deficits. Despite considerable recent progress, the specific synaptic function of neuroligin-1 (NL1) remains unclear. A current model proposes that NL1 acts exclusively to mature pre-existent synaptic connections in an activity-dependent manner. A second element of this activity-dependent maturation model is that an alternate molecule acts upstream of NL1 to initiate synaptic connections. SynCAM1 (SC1) is hypothesized to function in this capacity, though several uncertainties remain regarding SC1 function. Using overexpression and chronic pharmacological blockade of synaptic activity, we now demonstrate that NL1 is capable of robustly recruiting synapsin-positive terminals independent of synaptic maturation and activity in 2-week old primary hippocampal neuronal cultures. We further report that neither SC1 overexpression nor knockdown of endogenous SC1 impacts synapsin punctum densities, suggesting that SC1 is not a limiting factor of synapse initiation in maturing hippocampal neurons in vitro. Consistent with these findings, we observed profoundly greater recruitment of synapsin-positive presynaptic terminals by NL1 than SC1 in a mixed-culture assay of artificial synaptogenesis between primary neurons and heterologous cells. Collectively, our results contend multiple aspects of the proposed model of NL1 and SC1 function and motivate an alternative model whereby SC1 may mature synaptic connections forged by NL1. Supporting this model, we present evidence that combined NL1 and SC1 overexpression triggers excitotoxic neurodegeneration through SC1 signaling at synaptic connections initiated by NL1.


The small heat shock protein HSPB1 protects mice from sepsis.
In vitro studies have implicated the small heat shock protein HSPB1 in a range of physiological functions. However, its in vivo relevance is unclear as the phenotype of unstressed HSPB1-/- mice is unremarkable. To determine the impact of HSPB1 in injury, HSPB1-/- and wild type (WT) mice were subjected to cecal ligation and puncture, a model of polymicrobial sepsis. Ten-day mortality was significantly higher in HSPB1-/- mice following the onset of sepsis (65% vs. 35%). Ex vivo mechanical testing revealed that common carotid arteries from HSPB1-/- mice were more compliant than those in WT mice over pressures of 50-120 mm Hg. Septic HSPB1-/- mice also had increased peritoneal levels of IFN-γ and decreased systemic levels of IL-6 and KC. There were no differences in frequency of either splenic CD4+ or CD8+ T cells, nor were there differences in apoptosis in either cell type. However, splenic CD4+ T cells and CD8+ T cells from HSPB1-/- mice produced significantly less TNF and IL-2 following ex vivo stimulation. Systemic and local bacterial burden was similar in HSPB1-/- and WT mice. Thus while HSPB1-/- mice are uncompromised under basal conditions, HSPB1 has a critical function in vivo in sepsis, potentially mediated through alterations in arterial compliance and the immune response.


Sustainability, polysaccharide science, and bio-economy.
At the opening of the 2nd EPNOE conference the role and responsibility of polysaccharide scientists was reflected upon and placed in the context of actual global issues like the transition process towards "sustainable bio-economy". Difficulties in the chain of communication between the different parties involved and towards the wider public was addressed. The need for change in the relations between science and the public and to go beyond the horizon of the specialization was discussed. It was stated that polysaccharide science is one of the key sciences in those transitions.


Altered keratin biosynthesis follows inhibition of scale morphogenesis by hydrocortisone.
Hydrocortisone, administered onto the chorioallantoic membrane (CAM) of 7- to 10-day-old chick embryos, inhibits scale development, in a dose- and stage-dependent manner. The response is also region specific in that hydrocortisone treatment, at a specific dose and time, will completely block scutellate and interstitial scale development while leaving other scale types unaffected. Using histological, biochemical, and immunofluorescence techniques, we have shown that inhibition of scutellate scale morphogenesis prevents the subsequent formation of a beta stratum and alters expression of the alpha keratins. These data support the hypotheses that each avian scale type has its own distinctive temporal, morphological, and biochemical pattern of development; and in the case of scutellate scale development, hydrocortisone treatment alters keratin biosynthesis by interfering with earlier steps in morphogenesis.


The Fungal Mycobiota: Small Numbers, Large Impacts.
Fungi are an underappreciated component of the human gut microbiota. In this issue of Cell Host & Microbe, Wheeler et al. (2016) demonstrate that fungal dysbiosis induced by antifungal treatment or inoculation with typically rare fungi results in exaggerated immune responses, suggesting that fungal colonizers play key roles in immune homeostasis.


Clinical research from proposal to implementation: what every clinical investigator should know about the institutional review board.
The conduct of clinical trials is a complicated process involving a myriad of regulations and enforcement entities. To protect the rights and welfare of study participants, a system of oversight bodies called institutional review boards has been established in the US. This article describes how institutional review boards work and explains what clinical researchers need to know about federally mandated human subject protection requirements.


Label-free and rapid detection of ATP based on structure switching of aptamers.
In this work, an aptamer-based fluorescent strategy for label-free detection of ATP was developed by using Thioflavin T (ThT) as a fluorescence indicator, which can specifically bind with G-quadruplex DNAs to generate enhanced fluorescence intensity. In the absence of ATP, the folded structure of ATP aptamer allows the intercalation of ThT to produce strong fluorescence signal. However, upon ATP binding to the aptamer where ThT intercalated, the conformational change or distortion of the aptamer is large enough to cause much less intercalation of ThT and consequently drastic suppression of the fluorescence intensity. As such, the concentration of ATP could be identified very easily by observing fluorescence changes of this sensing system. This label-free assay could be accomplished very easily and quickly with a "mix-and-detect" detection method and exhibits high sensitivity to ATP with a detection limit of 33 nM in a wide range of 0.1-1000 μM. Furthermore, this proposed method is capable of detecting ATP in human serum and cell extracts. This method offers several advantages such as simplicity, rapidity, low cost, good stability and excellent selectivity, which make it hold great potential for the detection of ATP in bioanalytical and biological studies.


A ultra low level laboratory for nuclear test ban measurements.
The radionuclide laboratory at the Austrian Research Centers Seibersdorf (ATL03) was installed to support the international monitoring system for verification of the Comprehensive Nuclear-Test-Ban Treaty (CTBT and Text of the establishment of a Preparatory Commission for the Comprehensive Test-Ban-Treaty Organization, 1996). Therefore, the background of a high-purity germanium detection system has been reduced by developing a high sophisticated active and passive detector shielding. The entire system is encapsulated in an iron-castle and placed into a fall-out shelter. The final count rate, achieved over the energy interval from 40 to 2700 keV, amounts to 0.18 counts s(-1) kg(-1) (Ge).


Targeting EFNA1 suppresses tumor progression via the cMYC-modulated cell cycle and autophagy in esophageal squamous cell carcinoma.
Esophageal squamous cell carcinoma (ESCC) remains one of the most common causes of cancer death due to the lack of effective therapeutic options. New targets and the targeted drugs are required to be identified and developed. Highly expressed genes in ESCA were identified using the edgeR package from public datasets. Immunostaining assay verified the high expression level of EFNA1 in ESCC. CCK-8, colony formation and wound healing assays were performed to examine the role of EFNA1 and EPHA2 in ESCC progression. Cell cycle was analyzed by flow cytometry and autophagy activation was determined by autophagolysosome formation using transmission electron microscopy. The small molecule targeting to EFNA1 was identified by molecular docking and the anti-tumor effects were verified by in vitro and in vivo models with radiation treatment. EFNA1 was highly expressed in esophageal cancer and significantly associated with poor prognosis. Downregulation of EFNA1 remarkably inhibited cell proliferation and migration. Furthermore, decreased EFNA1 significantly suppressed the expression of cMYC along with its representative downstream genes involved in cell cycle, and activated autophagy. Similar effects on ESCC progression were obtained from knockdown of the corresponding receptor, EPHA2. The potential small molecule targeting to EFNA1, salvianolic acid A (SAA), could significantly suppress ESCC progression and increase the sensitivity to radiotherapy. We revealed that EFNA1 facilitated the ESCC progression via the possible mechanism of activating cMYC-modulated cell proliferation and suppressing autophagy, and identified SAA as a potential drug targeting EFNA1, providing new options for the future treatments for ESCC patients.


Influence of HIV infection and the use of antiretroviral therapy on selenium and selenomethionine concentrations and antioxidant protection.
The aim of the present study was to evaluate whether HIV infection and antiretroviral therapy (ART) use are associated with oxidative stress, concentrations of selenium and selenomethionine, and antioxidant protection. Individuals were classified as HIV negatives: control group (CG; n = 40); HIV positives: group 1 (G1; taking ART for >5 y, n = 40) and group 2 (G2; taking ART for <5 y, n = 40). Plasma and erythrocyte selenium, selenomethionine, glutathione (GSH), glutathione peroxidase activity (GPX), and malondialdehyde (MDA) were evaluated. Selenium deficiency (plasma selenium 45 μg/L) was not observed in any of the participants, and plasma selenium in CG (69.4 μg/L) was lower than in G1 and G2 (88.4 and 72.5 μg/L, respectively). G1 and G2 showed higher concentrations of MDA and GPX and lower concentration of GSH than CG. Multiple linear regression analysis indicated an association of MDA, GPX, and GSH with HIV status. CG participants showed higher concentrations of selenomethionine than G1 and G2 individuals and we observed a significant negative correlation between the concentration of selenomethionine and the use of ART. Prolonged ART use seems to increase the selenium in plasma, but influences the reduction of selenomethionine. HIV infection was associated with increased oxidative stress and appears to affect in protective activity of GPX. Finally, more studies are required to further address the importance of selenium and selenometabolites in the pathogenesis of infection and metabolism of HIV-positive individuals in prolonged use of ART.


Ex vivo analysis of desmoglein 1-responsive T-helper (Th) 1 and Th2 cells in patients with pemphigus foliaceus and healthy individuals.
Pemphigus foliaceus (PF) is a severe autoimmune bullous disorder, characterized by autoantibodies (autoAb) against desmoglein 1 (Dsg1). As T cells may be critical in the pathology of PF, the aim of the present study was to identify and characterize autoaggressive T-helper cells reactive to Dsg1 in PF patients and healthy individuals. Eight patients with the clinical diagnosis of PF and six HLA class II-matched healthy individuals were examined. By magnetic cell-sorting (MACS) cytokine-secretion assay, Dsg1-responsive T-helper (Th) 1 and Th2 cells were isolated and cloned by limiting dilution. The generated T-cell clones (TCC) were characterized regarding proliferative response, TCR Vbeta-chain usage, and cytokine profile upon in vitro stimulation with Dsg1. Both Dsg1-reactive Th1 and Th2 cells were detected in PF patients and controls at similar frequencies. A total of 15 Th1 and Th2 clones were isolated from patients and 27 TCC from healthy controls. Analysis of TCR Vbeta-chain usage of autoreactive T cells from both groups revealed no predominance of a specific Vbeta chain. Noteworthy, the isolated TCC showed a polarized Th1- or Th2-like phenotype upon in vitro culture and stable expression of Th1 or Th2 cytokines during long-term in vitro culture. In summary, our data demonstrate that T-cell autoreactivity against Dsg1 is not restricted to patients with PF. Moreover, both Th1 and Th2 cells were present in patients and healthy donors, suggesting that the loss of B-cell tolerance against Dsg1 in PF is not exclusively determined by the presence of autoaggressive T cells.


Uniform 40-µm-pore diameter precision templated scaffolds promote a pro-healing host response by extracellular vesicle immune communication.
Implanted porous precision templated scaffolds (PTS) with 40-µm spherical pores reduce inflammation and foreign body reaction (FBR) while increasing vascular density upon implantation. Larger or smaller pores, however, promote chronic inflammation and FBR. While macrophage (MØ) recruitment and polarization participates in perpetuating this pore-size-mediated phenomenon, the driving mechanism of this unique pro-healing response is poorly characterized. We hypothesized that the primarily myeloid PTS resident cells release small extracellular vesicles (sEVs) that induce pore-size-dependent pro-healing effects in surrounding T cells. Upon profiling resident immune cells and their sEVs from explanted 40-µm- (pro-healing) and 100-µm-pore diameter (inflammatory) PTS, we found that PTS pore size did not affect PTS resident immune cell population ratios or the proportion of myeloid sEVs generated from explanted PTS. However, quantitative transcriptomic assessment indicated cell and sEV phenotype were pore size dependent. In vitro experiments demonstrated the ability of PTS cell-derived sEVs to stimulate T cells transcriptionally and proliferatively. Specifically, sEVs isolated from cells inhabiting explanted 100 μm PTS significantly upregulated Th1 inflammatory gene expression in immortalized T cells. sEVs isolated from cell inhabiting both 40- and 100-μm PTS upregulated essential Treg transcriptional markers in both primary and immortalized T cells. Finally, we investigated the effects of Treg depletion on explanted PTS resident cells. FoxP3+ cell depletion suggests Tregs play a unique role in balancing T cell subset ratios, thus driving host response in 40-μm PTS. These results indicate that predominantly 40-µm PTS myeloid cell-derived sEVs affect T cells through a distinct, pore-size-mediated modality.


Using light and X-ray scattering to untangle complex neuronal orientations and validate diffusion MRI.
Disentangling human brain connectivity requires an accurate description of nerve fiber trajectories, unveiled via detailed mapping of axonal orientations. However, this is challenging because axons can cross one another on a micrometer scale. Diffusion magnetic resonance imaging (dMRI) can be used to infer axonal connectivity because it is sensitive to axonal alignment, but it has limited spatial resolution and specificity. Scattered light imaging (SLI) and small-angle X-ray scattering (SAXS) reveal axonal orientations with microscopic resolution and high specificity, respectively. Here, we apply both scattering techniques on the same samples and cross-validate them, laying the groundwork for ground-truth axonal orientation imaging and validating dMRI. We evaluate brain regions that include unidirectional and crossing fibers in human and vervet monkey brain sections. SLI and SAXS quantitatively agree regarding in-plane fiber orientations including crossings, while dMRI agrees in the majority of voxels with small discrepancies. We further use SAXS and dMRI to confirm theoretical predictions regarding SLI determination of through-plane fiber orientations. Scattered light and X-ray imaging can provide quantitative micrometer 3D fiber orientations with high resolution and specificity, facilitating detailed investigations of complex fiber architecture in the animal and human brain.


IL-10 gene modified dendritic cells inhibit T helper type 1-mediated alloimmune responses and promote immunological tolerance in diabetes.
Dendritic cells (DCs) have the potency to regulate the outcome of autoimmunity through the modulation of immune responses. The induction of antigen specific tolerance is critical for prevention and treatment of allograft rejection. In the present study, we transfected IL-10 gene into DCs and investigated their effect on inhibition of lymphocyte activity in vitro and induction of immune tolerance on islet allograft in mice. An IDDM C57BL/6 mouse model was induced by streptozotocin. The islet cells isolated from the BALB/c mice were transplanted into the kidney capules of the model mice followed by injection of IL-10 modified DCs (mDCs). The results showed that mDCs could significantly inhibit T lymphocyte proliferation mediated by allotype cells and induce its apoptosis, whereas, unmodified DCs (umDCs) could promote the murine lymphocyte proliferation markedly. The injection of mDCs could prolong the survival of allotype islet transplanted IDDM mice. The average plasma glucose (PG) level in mDCs treated mice returned to normal within 3 days and lasted for about 2 weeks. The rejection response in control mice occurred for 5 days after transplantation. The level of IFN-gamma was lower while IL-4 was higher in mDCs treated mice than that in umDCs treated mice, which indicated that Th1/Th2 deviation occurred. Our studies suggest that IL-10 gene modified DCs can induce the immune tolerance to islet graft and prolong survival of the recipients by the inhibiting of T cell proliferation in allotype mice.


Closure rates and patterns after light silicone oil tamponade for persistent full-thickness macular holes.
To report outcomes of re-vitrectomy using light silicone oil (SO) tamponade for persistent macular holes (MHs). We reviewed cases of patients with full-thickness MHs that underwent pars plana vitrectomy (PPV) with air/gas and were re-vitrectomized using light SO tamponade after primary non-closure (persistent MHs). Outcome measures included anatomic closure rates and patterns confirmed by optical coherence tomography (OCT) and changes in best-corrected visual acuity (BCVA). A total of 42 eyes of 41 patients with unsuccessful primary PPV with air/gas were included. After re-vitrectomy with light SO (1000-centistoke), 29 (69%) eyes demonstrated type 1 closure without neurosensory defects in OCT scans, whereas 9 eyes (21%) showed type 2 closure with persisting neurosensory defects. Available data (n = 21) showed a significant mean improvement of BCVA from 0.99 logMAR (SD 0.25, range 0.7-1.3) preoperatively to 0.74 logMAR (SD 0.42, range 0.2-1.5) postoperatively (p = 0.035). The treatment of persistent MHs with PPV and light SO tamponade resulted in high closure rates.


MACF1 gene structure: a hybrid of plectin and dystrophin.
Mammalian MACF1 (Macrophin1; previously named ACF7) is a giant cytoskeletal linker protein with three known isoforms that arise by alternative splicing. We isolated a 19.1-kb cDNA encoding a fourth isoform (MACF1-4) with a unique N-terminus. Instead of an N-terminal actin-binding domain found in the other three isoforms, MACF1-4 has eight plectin repeats. The MACF1 gene is located on human Chr 1p32, contains at least 102 exons, spans over 270 kb, and gives rise to four major isoforms with different N-termini. The genomic organization of the actin-binding domain is highly conserved in mammalian genes for both plectin and BPAG1. All eight plectin repeats are encoded by one large exon; this feature is similar to the genomic structure of plectin. The intron positions within spectrin repeats in MACF1 are very similar to those in the dystrophin gene. This demonstrates that MACF1 has characteristic features of genes for two classes of cytoskeletal proteins, i.e., plectin and dystrophin.


A combined variation of Palmaris longus and Flexor digitorum superficialis: Case report and review of literature.
The muscles of the anterior compartment of the forearm often exhibit anatomic variations. During dissection of the upper right limb of a preserved cadaver, morphological variations in the Palmaris longus and Flexor digitorum superficialis muscles were found. The Palmaris longus muscular fibers converged to a tendon that passed beneath the Flexor retinaculum, and inserted at the base of the middle phalanx of the fourth digit, replacing the tendon of Flexor digitorum superficialis. The Flexor digitorum superficialis was divided into two muscular heads: a digastric medial head giving tendons to the second and fifth digits, and a lateral head giving one tendon to the third digit. All these tendons were inserted in the respective digits by two bundles between which were located the tendon of the Flexor digitorum profundus muscle. Variations of flexor muscles must be documented because of their clinical significance and their potential use of these muscles in orthopaedic and reconstructive surgery.


Absolute frequency spectroscopy of CO2 lines at around 2.09 μm by combined use of an Er:fiber comb and a Ho:YLF amplifier.
The low-frequency tail of an octave-spanning supercontinuum (SC) generated by an Er:fiber comb is enhanced by a multipass Ho:YLF amplifier and used in a sum-frequency-generation scheme to obtain absolute referencing of a single-mode Tm-Ho:YAG laser tunable around 2.09 μm. By tuning the comb repetition frequency, the probing laser is scanned across the absorption lines of a CO(2) gas sample and highly accurate absorption profiles are measured. This approach can be readily scaled to any wavelength above ~2 μm.


Plasma membrane cholesterol is required for efficient pseudorabies virus entry.
Alphaherpesviruses comprise closely related viruses of man and animal, including herpes simplex virus, varicella-zoster virus and pseudorabies virus (PRV). Here, using methyl-beta-cyclodextrin and fluorescently tagged PRV, we directly show that depletion of cholesterol from the plasma membrane of host cells significantly reduces PRV entry. Cholesterol depletion did not reduce PRV attachment, but stalled virus particles at the plasma membrane before penetration of the cell. Cholesterol depletion results in destabilization of lipid raft microdomains in the plasma membrane, which have been shown before to be involved in efficient entry of different viruses. A significant fraction of PRV virions appears to localize juxtaposed to GM1, a lipid raft marker, during entry. Together, these data indicate that cholesterol and possibly cholesterol-rich lipid rafts may be important during PRV entry.


Optimizing and evaluating PCR-based pooled screening during COVID-19 pandemics.
Population screening played a substantial role in safely reopening the economy and avoiding new outbreaks of COVID-19. PCR-based pooled screening makes it possible to test the population with limited resources by pooling multiple individual samples. Our study compared different population-wide screening methods as transmission-mitigating interventions, including pooled PCR, individual PCR, and antigen screening. Incorporating testing-isolation process and individual-level viral load trajectories into an epidemic model, we further studied the impacts of testing-isolation on test sensitivities. Results show that the testing-isolation process could maintain a stable test sensitivity during the outbreak by removing most infected individuals, especially during the epidemic decline. Moreover, we compared the efficiency, accuracy, and cost of different screening methods during the pandemic. Our results show that PCR-based pooled screening is cost-effective in reversing the pandemic at low prevalence. When the prevalence is high, PCR-based pooled screening may not stop the outbreak. In contrast, antigen screening with sufficient frequency could reverse the epidemic, despite the high cost and the large numbers of false positives in the screening process.


Generation of human hybridomas producing migration inhibitory factor (MIF) and of murine hybridomas secreting monoclonal antibodies to human MIF.
Human T-cell hybridomas were established by hybridization of concanavalin A (Con A)-stimulated human peripheral blood T lymphocytes with cells from a 6-thioguanine-resistant, aminopterin-sensitive mutant line designated CEM-WH4, derived from the continuously growing human T cell line, CEM. High levels of MIF activity were demonstrated in the supernatants of two hybridoma lines, T-CEMA and T-CEMB but not of CEM-WH4 when stimulated with phorbol myristate acetate and phytohemagglutinin. In comparison, MIF derived from Con A-stimulated peripheral blood mononuclear cells showed 100 times less activity. Upon isoelectrofocusing, MIF activity of T-CEMB was found exclusively between pH 4.6 and 5.3 whereas MIF derived from T-CEMA showed heterogeneity with a major peak of MIF recovered at pH 4.6-5.3 and a minor peak at pH 2.4-3.3. These molecules, however, were all found to have an apparent MW of 68,000 and were resistant to trypsin. Most of these characteristics are in accordance with second day pH 3- and pH 5-MIF derived from peripheral blood mononuclear cells. When spleen cells from BALB/c mice immunized with T-CEMB-MIF were used to fuse with NS-1 mouse myeloma cells, nine hybridomas secreting antibodies to human MIF were obtained. Clone D112 which demonstrated the highest MIF-neutralizing activity was found to neutralize MIF derived from T-CEMA, peripheral blood mononuclear cells, and a T cell line, Mo.


Circumcision as treatment for Zoon's balanitis in an HIV-positive teenage patient.
In this report we describe a case of the Zoon's balanitis in a boy with HIV (AIDS B2). The clinical presentation, failure of topical treatment, cure by circumcision, and the histopathology findings are presented.


The use of steroids in cervicofacial infections in UK OMFS departments: a Maxillofacial Surgery Trainees Research Collaborative (MTReC) study.
Defining current inpatient management of cervicofacial infections is key to identifying strategies to optimise care. Steroid use is beneficial in peritonsillar and orbital abscess, but there are few studies of steroid use in odontogenic cervicofacial infections. Our previous research identified widespread variation in practice, necessitating a national audit to define current practice. A multicentre, trainee-led snapshot audit was carried out across 25 hospitals in the UK. Information recorded included demographic features, presentation, surgical and medical management, and steroid administration. One thousand two admissions were recorded. Fifty-five percent were male, with mean age 37.3 years (range 0-94 years). Steroids were administered to 357 patients (36%), 159 of whom received steroids on admission, and 254 peri-operatively. Patients who were prescribed steroids had significantly higher white cell count (12.4 vs 11.5), CRP (79.5 vs 57) and heart rate (94 vs 88) on admission. Justifications given for administration included swelling (74), dysphagia (21), and airway compromise (7). Three patients (0.8%) reported steroid-related side effects. This study uses the largest recorded cohort of cervicofacial infections to define current inpatient steroid administration. Our results suggest that steroids are targeted towards the most compromised 1/3 of patients. The incidence of steroid-related side effects was low. Defining the characteristics of patients currently receiving steroids will aid in determining whether steroid administration is an effective adjunct in treating odontogenic cervicofacial infection.


The NIH Science of Behavior Change Program: Looking Toward the Future.
The National Institutes of Health established the Science of Behavior Change (SOBC) program to promote basic research on the initiation, personalization, and maintenance of health behavior change. The SOBC Resource and Coordinating Center now leads and supports activities to maximize the creativity, productivity, scientific rigor, and dissemination of the experimental medicine approach and experimental design resources. Here, we highlight those resources, including the Checklist for Investigating Mechanisms in Behavior-change Research (CLIMBR) guidelines introduced in this special section. We describe the ways in which SOBC can be applied across a range of domains and contexts, and end by considering ways to extend SOBC's perspective and reach, so as to best promote behavior change linked with health, quality of life, and well-being.


Perceptual load does not modulate auditory distractor processing.
In vision, it is well established that the perceptual load of a relevant task determines the extent to which irrelevant distractors are processed. Much less research has addressed the effects of perceptual load within hearing. Here, we provide an extensive test using two different perceptual load manipulations, measuring distractor processing through response competition and awareness report. Across four experiments, we consistently failed to find support for the role of perceptual load in auditory selective attention. We therefore propose that the auditory system - although able to selectively focus processing on a relevant stream of sounds - is likely to have surplus capacity to process auditory information from other streams, regardless of the perceptual load in the attended stream. This accords well with the notion of the auditory modality acting as an 'early-warning' system as detection of changes in the auditory scene is crucial even when the perceptual demands of the relevant task are high.


"The Standard Procedure" for Investigation of Oral Neutrophils in Oral Diseases.
There is need of an objective "standard procedure" that is reliable and clinically applicable for estimating oral neutrophil content in relation to oral diseases. Forty-one patients with suspected oral candidosis (OC) and nine healthy controls with no oral mucosal disease were flushing with 10 ml mouth rinse (MR) (sterile phosphate-buffered saline) for 1 min. Aliquots were stored on different conditions to explore stability, storage, and fixation conditions for analysis by flow cytometry. The optimal storage and fixation condition for MR was by fixation 1 : 1 in 10% formalin and stored at 5°C. This procedure yielded stable results up to 7 days after collection. The ability of the optimized method to relate oral neutrophils to inflammation was demonstrated by the significantly higher number of neutrophils in patients with primary OC (p = 0.0334) compared to healthy controls. This method is rapid, reliable, and clinically applicable for establishing the content of oral neutrophils. We demonstrate increased density of oral neutrophils in the MR of patients with OC. The potential of the method is to be "the standard procedure" for investigation of the oral inflammation in patients with oral diseases as it is noninvasive and provides high stability, clinical relevance, and minimal handling.


T-lymphocyte subsets in the human lacrimal gland.
This study examines the microscopical appearance, location, distribution, subdivision and density of T-lymphocytes in the human lacrimal gland. Fourteen glands, 7 from either sex, were removed and frozen shortly after the donors' death, and processed for immunoperoxidase staining, utilizing a biotinavidin system and one of the following monoclonal antibodies: Anti-Leu-1 and -Leu-4, which recognize T-lymphocytes, anti-Leu-2a, which binds to suppressor/cytotoxic T-cells, and anti-Leu-3a + 3b, which recognizes helper/inducer cells. The T-lymphocytes in the human lacrimal gland were small to medium sized, mainly located in the interacinar tissue, often adjacent to an acinus or close to a collecting duct. The median number of each T-cell subset per defined field (0.086 mm2) at x 500 magnification was as follows: Males: Leu-1: 3.1, Leu-2a: 4.9, Leu-3a + 3b: 2.8, Leu-4: 4.5. Females: Leu-1: 4.3, Leu-2a: 5.0, Leu-3a + 3b: 3.7 and Leu-4:5.8. The sex difference was not statistically significant. The helper/suppressor cell ratio in the human lacrimal gland was 0.57 for males and 0.74 for females.


The role of HLA-G in cytokine homeostasis during early pregnancy complicated with maternal infections: a novel etiopathological approach to the neurodevelopmental understanding of schizophrenia.
Schizophrenia is perhaps the most enigmatic and tragic psychotic disorder with remarkable mortality and morbidity. Schizophrenia is complex and clinically a heterogeneous disorder. The etiological basis of schizophrenia ranges from autoimmune to neurodevelopmental hypothesis in one hand and involvement of different major gene segment with susceptibility loci on the other. Recently, neurodevelopmental hypothesis gained much impetus over the other domain. To support the neurodevelopmental basis, a number of investigations have shown that maternal infections during pregnancy increases the risk of the offspring developing schizophrenia and other neurodevelopmental disorders. The pathological mechanisms underlying this phenomenon is largely unknown. Many have suggested the involvement of different immune markers and shown that cytokines generated in response to maternal infection alter early brain development through their inflammatory activity. However, these findings have escaped discussion on various important issues related to cytokine homeostasis which depends on a large number of immune parameters including non-classical HLA-G molecules. Infections during early stages of pregnancy may alter cytokine regulation by disturbing the whole uterine immune milieu. To elucidate this issue, authors have tried to correlate the possible relationships between maternal infections and aberration of immune networking at the feto-maternal interface and their subsequent influence on the structural and functional abnormalities of the developing brain. The authors hypothesize that there exists a counter regulatory interaction among proinflammatory cytokines like TNF-alpha, HLA-G molecules and different immune cells like NK cells. We emphasize that HLA-G molecules are the novel immune players which maintain the immune homeostasis during early pregnancy in a manner that it can protect developing fetus from maternal immune attack. However, maternal infections may lead to the disturbance of HLA-G expression which in turn may fail to maintain its otherwise inhibitory potential to down regulate the detrimental inflammatory cytokines. Investigation on such interaction may unravel novel molecular mechanisms of neurodevelopmental basis of schizophrenia. Testing of our proposed hypothesis on animal models and on in vitro derived extravillous trophoblast cell lines holds promise of great insights to usher a new dimension of schizophrenia research and for developing new therapeutic strategies for better treatment and to adopt genetic prediction in schizophrenia management paradigm.


Global implementation of WHO's multimodal strategy for improvement of hand hygiene: a quasi-experimental study.
Health-care-associated infections are a major threat to patient safety worldwide. Transmission is mainly via the hands of health-care workers, but compliance with recommendations is usually low and effective improvement strategies are needed. We assessed the effect of WHO's strategy for improvement of hand hygiene in five countries. We did a quasi-experimental study between December, 2006, and December, 2008, at six pilot sites (55 departments in 43 hospitals) in Costa Rica, Italy, Mali, Pakistan, and Saudi Arabia. A step-wise approach in four 3-6 month phases was used to implement WHO's strategy and we assessed the hand-hygiene compliance of health-care workers and their knowledge, by questionnaire, of microbial transmission and hand-hygiene principles. We expressed compliance as the proportion of predefined opportunities met by hand-hygiene actions (ie, handwashing or hand rubbing). We assessed long-term sustainability of core strategy activities in April, 2010. We noted 21,884 hand-hygiene opportunities during 1423 sessions before the intervention and 23,746 opportunities during 1784 sessions after. Overall compliance increased from 51.0% before the intervention (95% CI 45.1-56.9) to 67.2% after (61.8-72.2). Compliance was independently associated with gross national income per head, with a greater effect of the intervention in low-income and middle-income countries (odds ratio [OR] 4.67, 95% CI 3.16-6.89; p<0.0001) than in high-income countries (2.19, 2.03-2.37; p<0.0001). Implementation had a major effect on compliance of health-care workers across all sites after adjustment for main confounders (OR 2.15, 1.99-2.32). Health-care-workers' knowledge improved at all sites with an increase in the average score from 18.7 (95% CI 17.8-19.7) to 24.7 (23.7-25.6) after educational sessions. 2 years after the intervention, all sites reported ongoing hand-hygiene activities with sustained or further improvement, including national scale-up. Implementation of WHO's hand-hygiene strategy is feasible and sustainable across a range of settings in different countries and leads to significant compliance and knowledge improvement in health-care workers, supporting recommendation for use worldwide. WHO, University of Geneva Hospitals, the Swiss National Science Foundation, Swiss Society of Public Health Administration and Hospital Pharmacists.


Identification of broad specificity P450CAM variants by primary screening against indole as substrate.
High-throughput screening of cytochrome P450CAM libraries, for their ability to oxidise indole to indigo and indirubin, has resulted in the identification of variants with activity towards the structurally unrelated substrate diphenylmethane.


Sub-wavelength terahertz imaging through optical rectification.
We record a sub-wavelength terahertz image of a caster sugar grain thanks to optical rectification in the sample excited with a femtosecond laser beam. The lateral spatial resolution of this technique is given by the laser spot size at the sample and here its measured value is 50 μm, i.e. ~λ/12. We give an estimation of the ultimate resolution that could be achieved with this method.


Phase-change material cooling blanket: A feasible cooling choice during transport after exercise-induced hyperthermia.
Exercise-induced hyperthermia preceding the onset of exertional heatstroke requires a rapid reduction in the body core temperature (Tcore) to ensure safety. In recent years, phase-change material (PCM) cooling devices have been increasingly used for rapid cooling after hyperthermia due to their superior capacity for heat absorption. This study aimed to evaluate the cooling performance and effectiveness of a PCM cooling blanket on heart rate (HR) and heart rate variability (HRV) recovery after exercise-induced hyperthermia. Randomized cross-over. The study participants were 12 male volunteers who were engaged in professional training and completed an endurance exercise for approximately 30 min in a hot and humid environment (temperature ≈ 30 °C; relative humidity ≈ 66%). The participants underwent a 30-min cooling trial after exercise, receiving either treatment with a PCM cooling blanket (PCM group) or natural cooling (CON group). The Tcore, HR, and HRV time-domain indices were used for analysis. The Tcore values were significantly lower in the PCM group during cooling. Reductions in the Tcore from precooling to 20 min of cooling were significantly greater in the PCM group than in the CON group. The HR in the PCM group was lower than that recorded in the CON group at 10 and 20 min of cooling. The reduction in HR during cooling from precooling was also significantly greater in the PCM group. HRV time-domain indices during cooling in the PCM group were significantly lower compared with the CON group while elevations in some HRV time-domain indices from precooling to postcooling were significantly greater in the PCM group than in the CON group. The PCM cooling blanket had good cooling performance and the ability to hasten recovery of both HR and HRV. It may serve as a feasible cooling choice during transport after exercise-induced hyperthermia.


[Oligopeptides in mechanisms of emotional reactions].
Oligopeptides participate in central mechanisms of emotional reactions. The oligopeptides are found, exhibiting at definite doses the activating and inhibitory effects on behavioral and vegetative manifestations of negative and positive emotional reactions.


Inhibition of hepatitis B virus production by modified 2',3'-dideoxy-thymidine and 2',3'-dideoxy-5-methylcytidine derivatives. In vitro and in vivo studies.
The effect of analogues of both 2',3'-dideoxy-3'-fluorothymidine (FddThd) [2',3'-dideoxy-3'-fluorouridine (FddUrd), 2',3'-dideoxy-3'-fluoro-5-chlorouridine (FddClUrd), 2',3'-dideoxy-3'- fluoro-5-bromouridine (FddBrUrd) and 2',3'-dideoxy-3'-fluoro-5-bromovinyluridine (FddBVUrd)] and 2',3'-dideoxy-3'-fluorocytidine (FddCyt) [2',3'-dideoxy-3'-fluoro-5-fluorocytidine (FddFCyt), 2',3'-dideoxy-3'-fluoro-5-chlorocytidine (FddClCyt), 2',3'-dideoxy-3'-fluoro-5-methylcytidine (FddMeCyt), 2',3'-dideoxy-3'-fluoro-5-ethylcytidine (FddEtCyt), 2',3'-dideoxy-3'-chloro-5-methylcytidine (ClddMeCyt), 2',3'-dideoxy-3'-amino-5-methylcytidine (AmddMeCyt), 2',3'-dideoxy-3'-azido-5- methylcytidine (AzddMeCyt) and arabinosyl-5-methylcytosine (AraMeCyt)] were tested for their potential antiviral activity in vitro using the human hepatoblastoma cell line, Hep G2 2.2.15, which was transfected with a vector containing hepatitis B virus (HBV). It was found that FddThd, FddMeCyt, FddEtCyt, ClddMeCyt, AmddMeCyt and AraMeCyt display cytostatic activity at concentrations (CD50 values) between 0.54 (FddMeCyt) and 3.93 microM (FddEtCyt), while FddUrd, FddClUrd, FddBrUrd, FddBVUrd, FddCyt, FddFCyt, FddClCyt and AzddMeCyt do not affect cell growth at concentrations of up to 25 microM. Among the thymidine analogues tested, FddThd is the most effective antiviral agent: at a concentration of 0.03 microM a more than 90% reduction of HBV DNA synthesis was measured. On the other hand, the antiviral indexes displayed by FddClUrd, FddBrUrd and FddBVUrd are higher than tht of FddThd; FddUrd was completely inactive. The most powerful antiviral agents in the group of cytidine analogues tested in vitro were FddMeCyt (more than 90% reduction of HBVDNA synthesis at 0.10 microM) and ClddMeCyt (0.10 microM); FddClCyt, FddEtCyt, AmddMeCyt and AraMeCyt were of intermediate activity. None of the negligible antiviral activity was determined for FddUrd, FddCyt, FddFCyt and AzddMeCyt. FddThd and FddMeCyt displayed in vivo an antiviral effect in the duck/duck HBV (DHBV) animal system. Administration of 10 or 20 mg/kg (total daily dose) of FddThd and 5 or 10 mg/kg of FddMeCyt (i.m. daily) to ducks infected with DHBV for 12 days blocked virus production. Termination of treatment with FddThd of infected animals led to reappearance of the virus in the serum though at lower levels. The in vitro and the in vivo data suggest that FddThd and FddMeCyt might be promising antiviral agents for the treatment of infection caused by HBV in humans.


Lymph node ratio-dependent prognosis stratification and postoperative radiotherapy utilization in T1-2N1 oral cavity carcinoma.
Pathological T1-2N1 oral cavity squamous cell carcinoma (pT1-2N1 OCSCC) is a setting with intermediate prognosis whilst without consensus regarding the utilization of postoperative radiotherapy (PORT). This study aimed to investigate the prognostic value of lymph node ratio (LNR) and to further examine its clinical validity for guiding PORT in pT1-2N1 OCSCC. OCSCC patients who received surgery between 2010 and 2015 with at least 6 lymph nodes dissection were extracted from the Surveillance, Epidemiology and End Results (SEER) database. Time-dependent receiver operating characteristic (ROC) analysis was used to identify the optimal cutoff of LNR. Multivariable Cox regression analysis was employed to assess the prognostic value of LNR. Impact of PORT was evaluated in respective subgroups stratified by LNR. A total of 870 OCSCC patients with pT1-2N1 diseases were eligible for analysis. The 5-year overall survival (OS) and disease-specific survival (DSS) was 57.2% and 67.9% respectively. Time-dependent ROC analyses for OS and DSS concordantly revealed 5.5% as the optimal cutoff of LNR. Significantly higher risks of death (HR = 1.610, 95% CI: 1.139-2.276) and disease-specific death (HR = 1.731, 95% CI: 1.101-2.723) were unveiled in patients with LNR > 5.5%. PORT related improvement on OS (5-year rate: 57.6% vs. 47.3%, p = 0.095) and DSS (5-year rate: 71.0% vs. 53.8%, p = 0.030) was only found in LNR > 5.5% subgroup. LNR > 5.5% is indicative of inferior outcome in pT1-2N1 OCSCC, warranting the utilization of PORT in this sub-setting.


Tumor Microenvironment in Mixed Neuroendocrine Non-Neuroendocrine Neoplasms: Interaction between Tumors and Immune Cells, and Potential Effects of Neuroendocrine Differentiation on the Tumor Microenvironment.
The tumor microenvironment is considered to play a pivotal role in various human malignancies. Neuroendocrine and non-neuroendocrine neoplasms are considered to have different tumor microenvironments. However, owing to differences in the systemic and/or local immune statuses, tumor microenvironments in different patients may be difficult to compare. Mixed neuroendocrine non-neuroendocrine neoplasms (MiNENs), although rare, could be useful for exploring the effects of neuroendocrine differentiation on the tumor microenvironment, because both neuroendocrine and non-neuroendocrine components are present in the same tumor. Here, we examined 33 cases of histologically confirmed MiNENs and evaluated the influence of neuroendocrine differentiation on the tumor microenvironment by comparing tumor-infiltrating lymphocytes, tumor-associated macrophages, and other relevant factors in the two components the same tumor. The immunoreactivity of those examined above was evaluated quantitatively. The values of vasohibin-1-positive density (p < 0.0001) and immunoreactivity (p < 0.0001) (representing the neoangiogenesis status) were significantly higher in neuroendocrine as compared to non-neuroendocrine areas of the same tumors. In addition, the Foxp3/CD8 (p = 0.0717) and the PD-1/CD8 ratios (p = 0.0176) (representing tumor immunity suppression) tend to increase in neuroendocrine carcinomas. Immunoreactivity of CD163, a marker of M2-like macrophages, was also higher in the neuroendocrine areas. Our findings indicate that neuroendocrine and non-neuroendocrine tumors differ from each other with respect to the characteristics of both tumor cells and the tumor microenvironment.


The use of a detailed video-based locomotor pattern analysis system to assess the functional reinnervation of denervated hind limb muscles.
Spinal cord injuries induce a critical loss of motoneurons followed by irreversible locomotor function impairment. Surgical approaches combined with neuroprotective agents effectively rescue the damaged motoneurons and improve locomotor function. Our aim was to develop a reliable method which is able to provide quantifiable and in-depth data on the locomotor recovery during skeletal muscle reinnervation. Sprague-Dawley rats underwent lumbar 4 ventral root avulsion and reimplantation followed by riluzole treatment in order to rescue the injured motoneurons of the damaged pool. Control animals were operated, but received no riluzole treatment. The locomotor pattern of the hind limb was recorded biweekly on a special runway equipped with high resolution and high speed digital cameras producing both lateral and rear views simultaneously. All together 12 parameters of the hind limb movement pattern were evaluated by measuring specific joint angles, footprints and gait parameters in single video frames. Four months after the operation Fast Blue, a fluorescent retrograde tracer was applied to the L4 spinal nerve in order to label the reinnervating motoneurons. Our results confirmed the sensitivity of our arrangement and established strong relationship between the functional improvement and the morphological reinnervation. Moreover, we developed a correction method to make the system tolerant to the differences in the weight, step duration and step length. There are no commercially available cheap, multi-parametric analysing equipment to characterise the gait in its complexity. Our system offers a modular, adaptable and expandable analysis on the reinnervation of the limb musculature in rodents.


Evaluation of a Modified SBAR Report to Physician Tool to Standardize Communication on Neonatal Transport.
SBAR (situation, background, assessment and recommendation) is a structured format for the effective communication of critically relevant information. This tool was developed as a generic template to provide structure to the communication of clinical information between health care providers. Neonatal transport often presents clinically stressful circumstances where concise and accurate information is required to be shared clearly between multidisciplinary health care providers. A modified SBAR communication tool was designed to facilitate structured communication between nonphysician bedside care providers operating from remote sites and physicians providing decision-making support at receiving care facilities. Prospective interventional study was designed to evaluate the reliability of a "SBAR report to physician tool" in sharing clinically relevant information between multidisciplinary care providers on neonatal transport. The study was conducted between 2011 and 2014 by a dedicated neonatal transport service based at McMaster Children's Hospital which provides care for approximately 500 infants in Southern Ontario annually. In the preintervention phase, 50 calls were randomly selected for the evaluation and 115 consecutively recorded transport calls following adoption of the reporting tool. The quality of calls prior to and after the intervention was assessed by reviewers independently. Inter-rater agreement was also assessed for both periods. Inter-rater agreement between raters was moderate to perfect in most components of the SBAR "report to the physician tool" except for the assessment component, which showed fair agreement during both preintervention and postintervention periods. There was an improvement in global score (primary outcome) with a mean difference of 0.95 (95% confidence interval [CI]: 0.77-1.14; p < 0.001) and in cumulative score with a mean difference of 8.55 (95% CI: 7.26-9.84; p < 0.001) in postintervention period. The use of the SBAR report to physician tool improved the quality of clinical information shared between nonphysician members of the neonatal transport team and neonatal transport physicians. · Long-Accurate and concise information sharing is crucial for decision-making in neonatal transport.. · Information sharing between multidisciplinary teams can be enhanced by using a commonly understood information sharing template.. · The SBAR report to physician tool improves the quality of information shared between multidisciplinary team members in neonatal transport..


Impact of aging on viral infections.
Older people are more susceptible to a variety of viral infections, including those that induce respiratory disease, resulting in higher morbidity and mortality than younger people. Aging impacts both innate and adaptive arms of the immune system to impair control of viral infections. This review will summarize key findings on how aging impacts immunity to viral infection.


Late postsplenectomy recurrence of thrombotic thrombocytopenic purpura responding to removal of accessory spleen.
The present report describes a patient who experienced recurrence of thrombotic thrombocytopenic purpura 10 years after the initial episode. The patient had been successfully treated with steroids and splenectomy and had complete clinical and hematologic remission. Thrombotic thrombocytopenic purpura recurred 10 years later and did not respond to steroids and plasmapheresis. The presence of an accessory spleen was demonstrated by technetium scanning. Surgical removal of the accessory spleen resulted again in prompt and complete recovery.


How do doctors choose treatment for older gynecological cancer patients? A Japanese Gynecologic Oncology Group survey of gynecologic oncologists.
The proportion of elderly Japanese people (age ≥ 65 years) is currently 27.7%, and the average life span of women is 87.14 years, both of which are unprecedented. In gynecologic cancer, evidence of treatment for the elderly is scarce, and treatment policies are determined by each facility. The aim of the present study was to investigate the status of treatment policies for elderly patients with gynecologic cancer. A web-based questionnaire regarding how treatment strategies are currently determined for elderly patients with gynecologic cancer was conducted on gynecologic oncologists to develop a tool for the objective evaluation of treatment policy decisions for elderly patients. The responses showed that 48% of the gynecologic oncologists were aware of comprehensive geriatric assessment (CGA), but only 6% had actually conducted CGA. Age, comorbidities, performance status, and pretreatment evaluations were regarded as important in determining the treatment strategy. Invasive treatments such as radical hysterectomy and para-aortic lymph node dissection tended to have age limits. These findings suggest that awareness of CGA is low in Japan, and that elderly people may not be given standard therapy, which highlights the importance of building on these findings by gathering further evidence and developing a new tool for predicting treatment outcomes for elderly patients with gynecologic cancer.


Combinatorial biosynthesis of lipopeptide antibiotics in Streptomyces roseosporus.
Daptomycin is a cyclic lipopeptide antibiotic produced by Streptomyces roseosporus. Cubicin (daptomycin-for-injection) was approved in 2003 by the FDA to treat skin and skin structure infections caused by Gram-positive pathogens. Daptomycin is particularly significant in that it represents the first new natural product antibacterial structural class approved for clinical use in three decades. The daptomycin gene cluster contains three very large genes (dptA, dptBC, and dptD) that encode the nonribosomal peptide synthetase (NRPS). The related cyclic lipopeptide A54145 has four NRPS genes (lptA, lptB, lptC, and lptD), and calcium dependent antibiotic (CDA) has three (cdaPS1, cdaPS2, and cdaPS3). Mutants of S. roseosporus containing deletions of one or more of the NRPS genes have been trans-complemented with dptA, dptBC, and dptD by inserting these genes under the control of the ermEp* promoter into separate conjugal cloning vectors containing phiC31 or IS117 attachment (attP int) sites; delivering the plasmids into S. roseosporus by conjugation from Escherichia coli; and inserting the plasmids site-specifically into the chromosome at the corresponding attB sites. This trans-complementation system was used to generate subunit exchanges with lptD and cdaPS3 and the recombinants produced novel hybrid molecules. Module exchanges at positions D: -Ala(8) and D: -Ser(11) in the peptide have produced additional novel derivatives of daptomycin. The approaches of subunit exchanges and module exchanges were combined with amino acid modifications of Glu at position 12 and natural variations in lipid side chain starter units to generate a combinatorial library of antibiotics related to daptomycin. Many of the engineered strains produced levels of novel molecules amenable to isolation and antimicrobial testing, and most of the compounds displayed antibacterial activities.


Analysis of mRNA synthesis following induction of the Escherichia coli SOS system.
Escherichia coli responds to impairment of DNA synthesis by inducing a system of DNA repair known as the SOS response. Specific genes are derepressed through proteolytic cleavage of their repressor, the lexA gene product. Cleavage in vivo requires functional RecA protein in a role not yet understood. We used mRNA hybridization techniques to follow the rapid changes that occur with induction in cells with mutations in the recA operator or in the repressor cleavage site. These mutations allowed us to uncouple the induction of RecA protein synthesis from its role in inducing the other SOS functions. Following induction with ultraviolet light, we observed increased rates of mRNA synthesis from five SOS genes within five minutes, maximum expression ten to 20 minutes later and then a later decline to near the initial rates. The presence of a recA operator mutation did not significantly influence these kinetics, whereas induction was fully blocked by an additional mutation in the repressor cleavage site. These experiments are consistent with activation of RecA protein preceding repressor cleavage and derepression of SOS genes. The results also suggest that the timing and extent of induction of individual SOS genes may be different.


Social Presence and Social Facilitation in Gambling: Some Comments on Yokomitsu et al. (2022).
This brief commentary adds to the recent study by paper by Yokomitsu, Kono and Takada (2022). Their study examined social presence in gambling by experimentally investigating the effects of the presence of other people on risky betting among high-risk gamblers. This commentary argues that the paper by Yokomitsu et al. provided a highly selective review on available studies and omitted many of the key studies in the area of social presence and social facilitation in which their findings could have been compared. The commentary also briefly outlines a number of studies that have I co-authored in this area over the past three decades using a variety of different methodologies (e.g., non-participant observation studies, experiments, data mining of account-based tracking data), none of which were mentioned by Yokomitsu et al. despite their clear relevance to this area.


Risk to pollinators from the use of chlorpyrifos in the United States.
CPY is an organophosphorus insecticide that is widely used in North American agriculture. It is non-systemic, comes in several sprayable and granular formulations,and is used on a number of high-acreage crops on which pollinators can forage,including tree fruits, alfalfa, corn, sunflower, and almonds. Bees (Apoidea) are the most important pollinators of agricultural crops in North America and were the main pollinators of interest in this risk assessment.The conceptual model identified a number of potential exposure pathways for pollinators, some more significant than others. CPY is classified as being highly toxic to honey bees by direct contact exposure. However, label precautions and good agricultural practices prohibit application of CPY when bees are flying and/or when flowering crops or weeds are present in the treatment area. Therefore, the risk of CPY to pollinators through direct contact exposure should be small. The main hazards for primary exposure for honey bees are dietary and contact exposure from flowers that were sprayed during application and remain available to bees after application. The main pathways for potential secondary exposure to CPY is through pollen and nectar brought to the hive by forager bees and the sublethal body burden of CPY carried on forager bees. Foraging for other materials, including water or propolis, does not appear to be an important exposure route. Since adult forager honey bees are most exposed, their protection from exposure via pollen, honey, and contact with plant surfaces is expected to be protective of other life stages and castes of honey bees.Tier- I approaches to estimate oral exposure to CPY through pollen and nectar/honey, the principle food sources for honey bees, suggested that CPY poses a risk to honey bees through consumption of pollen and nectar. However, a Tier-2 assessment of concentrations reported in pollen and honey from monitoring work in North America indicated there is little risk of acute toxicity from CPY through consumption of these food sources.Several models were also used to estimate upper-limit exposure of honey bees to CPY through consumption of water from puddles or dew. All models suggest that the risk of CPY is below the LOC for this pathway. Laboratory experiments with field-treated foliage, and semi-field and field tests with honey bees, bumble bees,and alfalfa leaf cutting bees indicate that exposure to foliage, pollen and/or nectar is hazardous to bees up to 3 d after application of CPY to a crop. Pollinators exposed to foliage, pollen or nectar after this time should be minimally affected.Several data gaps and areas of uncertainty were identified, which apply to CPYand other foliar insecticides. These primarily concern the lack of exposure and toxicological data on non-Apis pollinators. Overall, the rarity of reported bee kill incidents involving CPY indicates that compliance with the label precautions and good agricultural practice with the product is the norm in North American agriculture.Overall, we concluded that, provided label directions and good agricultural practices are followed, the use of CPY in agriculture in North America does not present an unacceptable risk to honeybees.


A global meta-analysis of crop yield and agricultural greenhouse gas emissions under nitrogen fertilizer application.
Nitrogen fertilization (NF) is one of the common practices to increase crop production worldwide over the past several decades. Nevertheless, unreasonable NF results in massive greenhouse gas (GHG) emissions, leading to climate change and global warming. Many studies have already reported the impact of NF on crop yield, global warming potential (GWP) and greenhouse gas intensity (GHGI), but the studies were limited to only some parameters. In this study, a total of 174 studies from 16 countries were collected and then a regression analysis was conducted to obtain the appropriate N fertilization rates that enhance crop yield while reducing GWP and GHGI. After that, a meta-analysis was performed to evaluate the effects of NF on crop yield, GHGI, GWP and GHG emissions and identify NF management strategies that benefit crop yield and maintain GWP. The results showed that the suitable N fertilization rate was 180, 150, 130 and 200 kg ha-1 for wheat, maize, rice and vegetables or industrial crops, respectively. Overall, NF resulted in positive effect size in crop yield (0.56) and negative effect size in GHGI (-0.14) compared to NNF. GWP showed positive effect size (0.37) due to an increase in N2O emissions (0.91) relative to NNF, which is higher than the increase of CH4 emissions (0.01) and CO2 emissions (0.22). It was recommended that split and banded application of urea or urea plus manure is employed for cereals (especially wheat) in the arid and semi-arid regions with medium-textured and neutral or alkaline soil.


Distribution of R-cognin and choline acetyltransferase in the ganglion cell layer of developing chick neural retina.
Retina cognin, a cell membrane glycoprotein which mediates cell-cell recognition and adhesion in vitro, is initially present throughout the retina and becomes confined to the ganglion cell layer at 14-15 days of embryogenesis. Within this layer it is found on membranes of virtually all ganglion and displaced amacrine cells, but not on membranes of retinal glial cells (Müller fibers) which traverse this layer. The distribution of cognin as determined by immunocytochemistry is described and compared with that of choline acetyltransferase. The significance of cognin as a possible address marker during development of neural retina is discussed.


Toxic metal tolerance in native plant species grown in a vanadium mining area.
Vanadium (V) has been extensively mined in China and caused soil pollution in mining area. It has toxic effects on plants, animals and humans, posing potential health risks to communities that farm and graze cattle adjacent to the mining area. To evaluate in situ phytoremediation potentials of native plants, V, chromium, copper and zinc concentrations in roots and shoots were measured and the bioaccumulation (BAF) and translocation (TF) efficiencies were calculated. The results showed that Setaria viridis accumulated greater than 1000 mg kg-1 V in its shoots and exhibited TF > 1 for V, Cr, Zn and BAF > 1 for Cu. The V accumulation amount in the roots of Kochia scoparia also surpassed 1000 mg kg-1 and showed TF > 1 for Zn. Chenopodium album had BAF > 1 for V and Zn and Daucus carota showed TF > 1 for Cu. Eleusine indica presented strong tolerance and high metal accumulations. S. viridis is practical for in situ phytoextractions of V, Cr and Zn and phytostabilisation of Cu in V mining area. Other species had low potential use as phytoremediation plant at multi-metal polluted sites, but showed relatively strong resistance to V, Cr, Cu and Zn toxicity, can be used to vegetate the contaminated soils and stabilise toxic metals in V mining area.


The effect of patented drug price on the share of new medicines across OECD countries.
The Government of Canada plans to implement new controls on the prices of patented drugs sold in Canada. The literature indicates that such controls delay drug launches. The Government of Canada, in its cost benefit analysis of the proposed regulatory changes, claims that they do not. To examine this claim, we use recent OECD country level data to estimate regression models of drug launches. These estimates suggest that higher drug list prices increase the number of launches of new medicines; the estimates are larger in the short term than in the longer term. If our estimates have a causal interpretation, then, consistent with the extant literature, drug list price reductions delay availability of new medicines in the OECD countries. We explore the implications of these findings.


Benefit evaluation in vitiligo treatment: development and validation of a patient-defined outcome questionnaire.
In addition to clinical efficacy, patient-defined benefit assessment has become an important outcome parameter. Thus far, no such instrument has been developed for vitiligo. Development and validation of a patient-defined benefit questionnaire in the therapy of vitiligo. Open questioning of 50 vitiligo patients generated 110 benefit items, which were converted into a 26-item questionnaire by a panel of dermatologists, psychologists and patients. This was used to record patient-defined needs prior to therapy and to evaluate patient benefit attained after therapy. A 'patient benefit index' (PBI) was formed on the basis of both. Validation was based on data of n = 1,023 vitiligo patients. The questionnaires were feasible in clinical practice, Cronbach's alpha (patient need questionnaire) was 0.94. The PBI showed convergent and discriminant validity with respect to quality of life and patient satisfaction. The PBI for vitiligo is a valid instrument, which is highly accepted in practice for recording patient-reported benefit.


Emerging from tabloid scandal: Dr. U.K. Mayo's Vegetable Vapor extends nitrous oxide's anesthetic duration.
Urial K. Mayo (1816-1900) was a successful Boston dentist who was plagued by personal scandal. In 1883 he patented extending the duration of nitrous-oxide anesthesia with an alcoholic tincture of hops and poppies.


Expression levels of CD47, CD35, CD55, and CD59 on red blood cells and signal-regulatory protein-alpha,beta on monocytes from patients with warm autoimmune hemolytic anemia.
Animal models have shown that CD47-deficient mice develop severe autoimmune hemolytic anemia (AIHA) because the binding of red blood cell (RBC) CD47 to signal-regulatory protein (SIRP-alpha) on macrophages contributes to the inhibition of phagocytosis. In contrast, complement-inhibitory proteins such as CD35, CD55, and CD59 may protect RBCs against the lysis by complement. With the use of flow cytometric analyses, the expression of CD47, CD35, CD55, and CD59 on RBCs and of SIRP-alpha,beta on peripheral monocytes of 36 patients with warm AIHA (wAIHA; 23 with active wAIHA, 13 with wAIHA in remission) and 20 healthy subjects was evaluated. The mean fluorescence intensities (MFIs) of the expression of CD47, CD35, CD55, and SIRP-alpha,beta of active wAIHA patients, wAIHA in remission, and healthy subjects were not statistically different. Patients with active wAIHA showed significantly lower CD59 expression on RBCs than healthy individuals (MFI, 512.5 +/- 59.6 vs. 553.7 +/- 36.6; p = 0.009), while the CD59 expression in patients with wAIHA in remission was not significantly different from that of healthy controls (MFI, 538.4 +/- 48.3 vs. 553.7 +/- 36.6; p > 0.05). The expression of CD59 on RBCs of 3 patients who died from the wAIHA was lower than that seen on RBCs of healthy controls (MFI, 433.6 +/- 69.6 vs. 553.74 +/- 36.6; p = 0.0001). Our data show that the expression of CD47 on RBCs and SIRP-alpha,beta on monocytes of patients with wAIHA is not different from that seen in healthy individuals. In addition, we detected that patients with active wAIHA present low expression of CD59 and normal expression of CD35 and CD55 on their RBCs. Complement-regulatory proteins may play an important role in protecting RBC destruction through the activation of complement.


Changes in brain serotonin levels affect leukocytes, lymphocytes, T-cell subpopulations and natural killer cell activity in rats.
The intraventricular injection of 5,6-dihydroxytryptamine into desipramine-treated rats caused a marked reduction in serotonin levels 5 and 10 days after treatment without significantly affecting catecholamine levels. In these animals, total leukocytes were increased and lymphocytes decreased. The number of helper T-cells (W3/25+) was significantly decreased after 5 days, whereas the numbers of total T-cells (W3/13+) and suppressor T-cells (MRC OX8) were increased after 10 days. The percentages of total T-cells, helper T-cells and suppressor T-cells were significantly reduced after 5 days. Natural killer cell (NK) activity was markedly reduced after 5 days. This finding suggests that central serotonin-containing neurons might be involved in the regulation of certain parts of the peripheral immune system.


[Recommendations on the safety of patients for socio-health centers: systematic review].
We did a systematic review to find recommendations on patient safety oriented toward improving the quality of care in nursing homes, residential facilities, housing for the elderly and long-term care facilities, among others. One hundred and thirty-four articles were selected in MEDLINE, EMBASE and CINAHL up to October 2012. Of these, 17 met inclusion criteria and 5 studies were added in the secondary search for further detailed analysis. Few studies with high or very high level of scientific evidence on the scale SIGN were identified. Analyzed studies focused primarily on nursing staff. Most of the recommendations are oriented toward medication-related issues, staff training, pressure ulcers or falls, adherence to guidelines and protocols and topics referred to organizational culture.


Bone loss in males with ankylosing spondylitis: its relation to sex hormone levels.
Osteoporosis is a common complication of ankylosing spondylitis (AS). The aim of the present study was to assess the association of sex hormone levels with bone loss in males with AS. A cross-sectional study. Patients The study group consisted of 58 male patients with primary AS. The mean age was 38.2 years (range 18-59 years). Bone mineral density (BMD) was measured in the left proximal femur. Serum FSH, LH, total testosterone (T), dehydroepiandrosterone sulfate (DHEAS), oestradiol (E2) and progesterone levels were measured. Bone loss was observed in 54.5% of cases at the femoral neck and in 52.3% of cases at the trochanter and total hip. Bone loss was found in 40 (69%) cases in at least one of these three regions. Serum DHEAS was low in 12 (30.8%) of the cases with bone loss, and one (5.9%) of those without (P = 0.043). The ratio of serum T/DHEAS was higher in those with bone loss (5.24 +/- 3.70) than in those without (3.58 +/- 3.16) (P = 0.026). The results showed that bone loss might be related to low serum DHEAS levels in males with AS.