p53, miR-34a and EMP1-Newly Identified Targets of TFF3 Signaling in Y79 Retinoblastoma Cells.
Trefoil factor family peptide 3 (TFF3) is supposed to have tumor suppressive functions in retinoblastoma (RB), but the functional pathway is not completely understood. In the study presented, we investigated the downstream pathway of TFF3 signaling in Y79 RB cells. Results from pG13-luciferase reporter assays and western blot analyses indicate induced p53 activity with an upregulation of miR-34a after TFF3 overexpression. Expression levels of the predicted miR-34a target epithelial membrane protein 1 (EMP1) are reduced after TFF3 overexpression. As revealed by WST-1 assay, BrdU, and DAPI cell counts viability and proliferation of Y79 cells significantly decrease following EMP1 knockdown, while apoptosis levels significantly increase. Opposite effects on Y79 cells' growth could be shown after EMP1 overexpression. Caspase assays showed that EMP1 induced apoptosis after overexpression is at least partially caspase-3/7 dependent. Colony formation and soft agarose assays, testing for anchorage independent growth, revealed that EMP1 overexpressing Y79 cells have a significantly higher ability to form colonies. In in ovo chicken chorioallantoic membrane (CAM) assays inoculated EMP1 overexpressing Y79 cells form significantly larger CAM tumors. Moreover, miR-34a overexpression increases sensitivity of Y79 cells towards RB chemotherapeutics, however, without involvement of EMP1. In summary, the TFF3 signaling pathway in Y79 RB cells involves the activation of p53 with downstream induction of miR-34a and subsequent inhibition of EMP1.


Cross-site and cross-platform variability of automated patch clamp assessments of drug effects on human cardiac currents in recombinant cells.
Automated patch clamp (APC) instruments enable efficient evaluation of electrophysiologic effects of drugs on human cardiac currents in heterologous expression systems. Differences in experimental protocols, instruments, and dissimilar site procedures affect the variability of IC50 values characterizing drug block potency. This impacts the utility of APC platforms for assessing a drug's cardiac safety margin. We determined variability of APC data from multiple sites that measured blocking potency of 12 blinded drugs (with different levels of proarrhythmic risk) against four human cardiac currents (hERG [IKr], hCav1.2 [L-Type ICa], peak hNav1.5, [Peak INa], late hNav1.5 [Late INa]) with recommended protocols (to minimize variance) using five APC platforms across 17 sites. IC50 variability (25/75 percentiles) differed for drugs and currents (e.g., 10.4-fold for dofetilide block of hERG current and 4-fold for mexiletine block of hNav1.5 current). Within-platform variance predominated for 4 of 12 hERG blocking drugs and 4 of 6 hNav1.5 blocking drugs. hERG and hNav1.5 block. Bland-Altman plots depicted varying agreement across APC platforms. A follow-up survey suggested multiple sources of experimental variability that could be further minimized by stricter adherence to standard protocols. Adoption of best practices would ensure less variable APC datasets and improved safety margins and proarrhythmic risk assessments.


Lead speciation in indoor dust: a case study to assess old paint contribution in a Canadian urban house.
Residents in older homes may experience increased lead (Pb) exposures due to release of lead from interior paints manufactured in past decades, especially pre-1960s. The objective of the study was to determine the speciation of Pb in settled dust from an urban home built during WWII. X-ray absorption near-edge structure (XANES) and micro-X-ray diffraction (XRD) analyses were performed on samples of paint (380-2,920 mg Pb kg(-1)) and dust (200-1,000 mg Pb kg(-1)) collected prior to renovation. All dust samples exhibited a Pb XANES signature similar to that of Pb found in paint. Bulk XANES and micro-XRD identified Pb species commonly found as white paint pigments (Pb oxide, Pb sulfate, and Pb carbonate) as well as rutile, a titanium-based pigment, in the <150 μm house dust samples. In the dust fraction <36 μm, half of the Pb was associated with the Fe-oxyhydroxides, suggesting additional contribution of outdoor sources to Pb in the finer dust. These results confirm that old paints still contribute to Pb in the settled dust for this 65-year-old home. The Pb speciation also provided a clearer understanding of the Pb bioaccessibility: Pb carbonate > Pb oxide > Pb sulfate. This study underscores the importance of taking precautions to minimize exposures to Pb in house dust, especially in homes where old paint is exposed due to renovations or deterioration of painted surfaces.


Virtual Reality Objectifies the Diagnosis of Psychiatric Disorders: A Literature Review.
To date, a diagnosis in psychiatry is largely based on a clinical interview and questionnaires. The retrospective and subjective nature of these methods leads to recall and interviewer biases. Therefore, there is a clear need for more objective and standardized assessment methods to support the diagnostic process. The introduction of virtual reality (VR) creates the possibility to simultaneously provoke and measure psychiatric symptoms. Therefore, VR could contribute to the objectivity and reliability in the assessment of psychiatric disorders. In this literature review, we will evaluate the assessment of psychiatric disorders by means of VR environments. First, we investigate if these VR environments are capable of simultaneously provoking and measuring psychiatric symptoms. Next, we compare these measures with traditional diagnostic measures. We performed a systematic search using PubMed, Embase, and Psycinfo; references of selected articles were checked for eligibility. We identified studies from 1990 to 2016 on VR used in the assessment of psychiatric disorders. Studies were excluded if VR was used for therapeutic purposes, if a different technique was used, or in case of limitation to a non-clinical sample. A total of 39 studies were included for further analysis. The disorders most frequently studied included schizophrenia (n = 15), developmental disorders (n = 12), eating disorders (n = 3), and anxiety disorders (n = 6). In attention-deficit hyperactivity disorder, the most comprehensive measurement was used including several key symptoms of the disorder. Most of the studies, however, concerned the use of VR to assess a single aspect of a psychiatric disorder. In general, nearly all VR environments studied were able to simultaneously provoke and measure psychiatric symptoms. Furthermore, in 14 studies, significant correlations were found between VR measures and traditional diagnostic measures. Relatively small clinical sample sizes were used, impeding definite conclusions. Based on this review, the innovative technique of VR shows potential to contribute to objectivity and reliability in the psychiatric diagnostic process.


Avoidable factors that contribute to the complications of ventriculoperitoneal shunt in childhood hydrocephalus.
A personal series of 80 consecutive children who underwent ventriculoperitoneal shunting for hydrocephalus has been followed up for a period of from 6 months to 6.5 years. Complications occurred in 8 patients; these included partial occlusion of the ventricular catheter in 2, infection in 2, peritoneal catheter-valve disconnection with migration of the catheter into the peritoneal cavity in 2, fracture of the peritoneal catheter just below the valve in 1, and failure of the peritoneal cavity to absorb cerebrospinal fluid in another. Analysis of the results in the present series showed that the complications result from the technique. The operative procedures responsible for a low complication rate in the present series are described. It is concluded that to avoid shunt complications, attention must be paid to the following factors: meticulous asepsis; good surgical technique, including testing of the shunt system to make certain that the correct opening pressure is present; elimination of contact between the shunt system and the patient's skin; placement of the valve under a pericranial flap; positioning the tip of the ventricular catheter just in front of the foramen of Monro and that of the peritoneal catheter in the pelvic peritoneal cavity.


Neuraminidase A-Exposed Galactose Promotes Streptococcus pneumoniae Biofilm Formation during Colonization.
Streptococcus pneumoniae is an opportunistic pathogen that colonizes the nasopharynx. Herein we show that carbon availability is distinct between the nasopharynx and bloodstream of adult humans: glucose is absent from the nasopharynx, whereas galactose is abundant. We demonstrate that pneumococcal neuraminidase A (NanA), which cleaves terminal sialic acid residues from host glycoproteins, exposed galactose on the surface of septal epithelial cells, thereby increasing its availability during colonization. We observed that S. pneumoniae mutants deficient in NanA and β-galactosidase A (BgaA) failed to form biofilms in vivo despite normal biofilm-forming abilities in vitro Subsequently, we observed that glucose, sucrose, and fructose were inhibitory for biofilm formation, whereas galactose, lactose, and low concentrations of sialic acid were permissive. Together these findings suggested that the genes involved in biofilm formation were under some form of carbon catabolite repression (CCR), a regulatory network in which genes involved in the uptake and metabolism of less-preferred sugars are silenced during growth with preferred sugars. Supporting this notion, we observed that a mutant deficient in pyruvate oxidase, which converts pyruvate to acetyl-phosphate under non-CCR-inducing growth conditions, was unable to form biofilms. Subsequent comparative transcriptome sequencing (RNA-seq) analyses of planktonic and biofilm-grown pneumococci showed that metabolic pathways involving the conversion of pyruvate to acetyl-phosphate and subsequently leading to fatty acid biosynthesis were consistently upregulated during diverse biofilm growth conditions. We conclude that carbon availability in the nasopharynx impacts pneumococcal biofilm formation in vivo Additionally, biofilm formation involves metabolic pathways not previously appreciated to play an important role.


Signaling Interplay between Bone Marrow Adipose Tissue and Multiple Myeloma cells.
In the year 2000, Hanahan and Weinberg (1) defined the six Hallmarks of Cancer as: self-sufficiency in growth signals, evasion of apoptosis, insensitivity to antigrowth mechanisms, tissue invasion and metastasis, limitless replicative potential, and sustained angiogenesis. Eleven years later, two new Hallmarks were added to the list (avoiding immune destruction and reprograming energy metabolism) and two new tumor characteristics (tumor-promoting inflammation and genome instability and mutation) (2). In multiple myeloma (MM), a destructive cancer of the plasma cell that grows predominantly in the bone marrow (BM), it is clear that all these hallmarks and characteristics are in play, contributing to tumor initiation, drug resistance, disease progression, and relapse. Bone marrow adipose tissue (BMAT) is a newly recognized contributor to MM oncogenesis and disease progression, potentially affecting MM cell metabolism, immune action, inflammation, and influences on angiogenesis. In this review, we discuss the confirmed and hypothetical contributions of BMAT to MM development and disease progression. BMAT has been understudied due to technical challenges and a previous lack of appreciation for the endocrine function of this tissue. In this review, we define the dynamic, responsive, metabolically active BM adipocyte. We then describe how BMAT influences MM in terms of: lipids/metabolism, hypoxia/angiogenesis, paracrine or endocrine signaling, and bone disease. We then discuss the connection between BMAT and systemic inflammation and potential treatments to inhibit the feedback loops between BM adipocytes and MM cells that support MM progression. We aim for researchers to use this review to guide and help prioritize their experiments to develop better treatments or a cure for cancers, such as MM, that associate with and may depend on BMAT.


Randomized controlled trial of meat compared with multimicronutrient-fortified cereal in infants and toddlers with high stunting rates in diverse settings.
Improved complementary feeding is cited as a critical factor for reducing stunting. Consumption of meats has been advocated, but its efficacy in low-resource settings has not been tested. The objective was to test the hypothesis that daily intake of 30 to 45 g meat from 6 to 18 mo of age would result in greater linear growth velocity and improved micronutrient status in comparison with an equicaloric multimicronutrient-fortified cereal. This was a cluster randomized efficacy trial conducted in the Democratic Republic of Congo, Zambia, Guatemala, and Pakistan. Individual daily portions of study foods and education messages to enhance complementary feeding were delivered to participants. Blood tests were obtained at trial completion. A total of 532 (86.1%) and 530 (85.8%) participants from the meat and cereal arms, respectively, completed the study. Linear growth velocity did not differ between treatment groups: 1.00 (95% CI: 0.99, 1.02) and 1.02 (95% CI: 1.00, 1.04) cm/mo for the meat and cereal groups, respectively (P = 0.39). From baseline to 18 mo, stunting [length-for-age z score (LAZ) <-2.0] rates increased from ~33% to nearly 50%. Years of maternal education and maternal height were positively associated with linear growth velocity (P = 0.0006 and 0.003, respectively); LAZ at 6 mo was negatively associated (P < 0.0001). Anemia rates did not differ by group; iron deficiency was significantly lower in the cereal group. The high rate of stunting at baseline and the lack of effect of either the meat or multiple micronutrient-fortified cereal intervention to reverse its progression argue for multifaceted interventions beginning in the pre- and early postnatal periods.


Severe malarial anemia: innate immunity and pathogenesis.
Greater than 80% of malaria-related mortality occurs in sub-Saharan Africa due to infections with Plasmodium falciparum. The majority of P. falciparum-related mortality occurs in immune-naïve infants and young children, accounting for 18% of all deaths before five years of age. Clinical manifestations of severe falciparum malaria vary according to transmission intensity and typically present as one or more life-threatening complications, including: hyperparasitemia; hypoglycemia; cerebral malaria; severe malarial anemia (SMA); and respiratory distress. In holoendemic transmission areas, SMA is the primary clinical manifestation of severe childhood malaria, with cerebral malaria occurring only in rare cases. Mortality rates from SMA can exceed 30% in pediatric populations residing in holoendemic transmission areas. Since the vast majority of the morbidity and mortality occurs in immune-naïve African children less than five years of age, with SMA as the primary manifestation of severe disease, this review will focus primarily on the innate immune mechanisms that govern malaria pathogenesis in this group of individuals. The pathophysiological processes that contribute to SMA involve direct and indirect destruction of parasitized and non-parasitized red blood cells (RBCs), inefficient and/or suppression of erythropoiesis, and dyserythropoiesis. While all of these causal etiologies may contribute to reduced hemoglobin (Hb) concentrations in malaria-infected individuals, data from our laboratory and others suggest that SMA in immune-naïve children is characterized by a reduced erythropoietic response. One important cause of impaired erythroid responses in children with SMA is dysregulation in the innate immune response. Phagocytosis of malarial pigment hemozoin (Hz) by monocytes, macrophages, and neutrophils is a central factor for promoting dysregulation in innate inflammatory mediators. As such, the role of P. falciparum-derived Hz (PfHz) in mediating suppression of erythropoiesis through its ability to cause dysregulation in pro- and anti-inflammatory cytokines, growth factors, chemokines, and effector molecules is discussed in detail. An improved understanding of the etiological basis of suppression of erythropoietic responses in children with SMA may offer the much needed therapeutic alternatives for control of this global disease burden.


Inhibitory effects of spinal propofol on the responses of spinal dorsal horn neurons in normal rats.
Spinal dorsal horn neurons play an important role in the processing of sensory information and are also targets of modulation by both endogenous and exogenous drugs. Propofol is an intravenous anesthetic and whether it has direct modulatory actions on sensory neuronal responses of the spinal cord dorsal horn has not been well studied. In the present study, a single dose (0.5 micromol) of propofol dissolved in dimethyl sulfoxide (DMSO) was directly applied onto the dorsal surface of the spinal cord and its effect was evaluated in 25 wide-dynamic-range (WDR) neurons and 10 low-threshold mechanoreceptive (LTM) neurons by using extracellular single unit recording technique in sodium pentobarbital anesthetized rats. Compared with the DMSO treatment, propofol produced a significant inhibition of WDR neuronal activity evoked by both noxious heat (45, 47, 49 or 53 degrees C, 15 s) and mechanical (pinch, 10 s) stimuli applied to their cutaneous receptive fields (cRF) on the ipsilateral hind paw skin. To investigate whether propofol exerts a modulatory effect on non-nociceptive afferent-mediated activity, the responses of WDR or LTM neurons to non-noxious brush and pressure were also evaluated. The non-noxious mechanically-evoked responses of both WDR and LTM neurons were significantly suppressed by propofol. The present results indicate that propofol has direct actions on the dorsal horn neurons of the spinal cord in rats. However, since both non-nociceptive and nociceptive afferent-mediated activity can be suppressed, the spinal effects of propofol are not likely to be specifically associated with anti-nociception.


Vascular endothelial growth factor expression in early stage ovarian carcinoma.
Tumor angiogenesis is essential for solid tumor growth. Yet, the importance of any particular factor in neoplastic proliferation is poorly defined. This study examines the clinical significance of increased expression of one of the angiogenic factors, vascular endothelial growth factor (VEGF), in early stage ovarian carcinoma. Tumor specimens from 68 patients with International Federation of Gynecology and Obstetrics Stage I and II ovarian carcinoma were evaluated for VEGF expression. Antisense and corresponding sense (control) RNA probes were transcribed from the pCRII construct (Invitrogen, San Diego, CA), which contained human VEGF cDNA. The antisense probe was designed to include a highly conserved region of the VEGF coding sequence and thus detect all known variants. After in situ hybridization, sections were assessed for overexpression of VEGF. Twenty-nine of the tumor samples overexpressed VEGF, whereas 39 specimens did not. In patients whose tumors demonstrated elevated VEGF expression, 25% were without evidence of disease recurrence at last follow-up. In contrast, 75% of the patients whose tumors did not overexpress VEGF were without evidence of disease at last follow-up (P < 0.001). Median disease free survival for the VEGF positive group was 22 months, compared with > 108 months for the VEGF negative group (P < 0.001). When borderline tumors were excluded from the survival analysis, median disease free survival for the VEGF positive group was 18 months, compared with >120 months for the VEGF negative group (P < 0.001). Other possible prognostic variables had minimal impact on survival; these included age, stage, grade, cytology, and tumor size (P > 0.05). Assignment to a high risk group, as defined by the Gynecologic Oncology Group of the National Cancer Institute, was somewhat predictive of a shorter relapse free interval (P = 0.056). In a multivariate analysis, however, only elevated VEGF expression was associated with poorer survival. In this analysis, patients with early stage ovarian carcinoma with increased VEGF expression had a poorer prognosis. Further study of VEGF may ultimately lead to identification of patients with high risk lesions whose tumor biology portends a worse prognosis and who therefore may benefit from aggressive adjuvant therapy.


Effect of cholesterol on the structure of a phospholipid bilayer.
Cholesterol plays an important role in regulating the properties of phospholipid membranes. To obtain a detailed understanding of the lipid-cholesterol interactions, we have developed a mesoscopic water-lipid-cholesterol model. In this model, we take into account the hydrophobic-hydrophilic interactions and the structure of the molecules. We compute the phase diagram of dimyristoylphosphatidylcholine-cholesterol by using dissipative particle dynamics and show that our model predicts many of the different phases that have been observed experimentally. In quantitative agreement with experimental data our model also shows the condensation effect; upon the addition of cholesterol, the area per lipid decreases more than one would expect from ideal mixing. Our calculations show that this effect is maximal close to the main-phase transition temperature, the lowest temperature for which the membrane is in the liquid phase, and is directly related to the increase of this main-phase transition temperature upon addition of cholesterol. We demonstrate that no condensation is observed if we slightly change the structure of the cholesterol molecule by adding an extra hydrophilic head group or if we decrease the size of the hydrophobic part of cholesterol.


Nestin is expressed in the basal/myoepithelial layer of the mammary gland and is a selective marker of basal epithelial breast tumors.
Transcriptional profiling has identified five breast cancer subtypes, of which the basal epithelial is most aggressive and correlates with poor prognosis. These tumors display a high degree of cellular heterogeneity and lack established molecular targets, such as estrogen receptor-alpha, progesterone receptor, and Her2 overexpression, indicating a need for definitive diagnostic markers. We present evidence that nestin, a previously described marker of regenerative cells in diverse tissues, is expressed in the regenerative compartment of the normal human mammary gland. Colocalization studies indicate two distinct populations of mammary epithelia that express nestin: one expressing cytokeratin 14 (CK14) and DeltaN-p63 and another expressing desmin. Immunohistochemical analysis indicates that DeltaN-p63 and nestin are coordinately expressed during pregnancy in the murine mammary gland. In the embryonal carcinoma cell line NT2/D1, ectopic DeltaN-p63-alpha disrupts retinoic acid-induced differentiation, thereby preserving expression of nestin; however, small interfering RNA-mediated ablation of nestin is insufficient to promote differentiation, indicating that whereas nestin may identify cells within the regenerative compartment of the mammary gland, it is insufficient to block differentiation and preserve replicative capacity. Immunohistochemical analysis of basal epithelial breast tumors, including those shown to carry BRCA1 mutations, indicates robust expression of nestin and CK14, punctate expression of p63, and low to undetectable levels of desmin expression. Nestin was not detected in other breast cancer subtypes, indicating selectivity for basal epithelial breast tumors. These studies identify nestin as a selective marker of the basal breast cancer phenotype, which displays features of mammary progenitors.


Role of Cationic Side Chains in the Antimicrobial Activity of C18G.
Antimicrobial peptides (AMPs) have been an area of great interest, due to the high selectivity of these molecules toward bacterial targets over host cells and the limited development of bacterial resistance to these molecules throughout evolution. The peptide C18G has been shown to be a selective, broad spectrum AMP with a net +8 cationic charge from seven lysine residues in the sequence. In this work, the cationic Lys residues were replaced with other natural or non-proteinogenic cationic amino acids: arginine, histidine, ornithine, or diaminopropionic acid. These changes vary in the structure of the amino acid side chain, the identity of the cationic moiety, and the pKa of the cationic group. Using a combination of spectroscopic and microbiological methods, the influence of these cationic groups on membrane binding, secondary structure, and antibacterial activity was investigated. The replacement of Lys with most other cationic residues had, at most, 2-fold effects on minimal inhibitory concentration against a variety of Gram-positive and Gram-negative bacteria. However, the peptide containing His as the cationic group showed dramatically reduced activity. All peptide variants retained the ability to bind lipid vesicles and showed clear preference for binding vesicles that contained anionic lipids. Similarly, all peptides adopted a helical conformation when bound to lipids or membrane mimetics, although the peptide containing diaminopropionic acid exhibited a decreased helicity. The peptides exhibited a wider variety of activity in the permeabilization of bacterial membranes, with peptides containing Lys, Arg, or Orn being the most broadly active. In all, the antibacterial activity of the C18G peptide is generally tolerant to changes in the structure and identity of the cationic amino acids, yielding new possibilities for design and development of AMPs that may be less susceptible to immune and bacterial recognition or in vivo degradation.


Candida orthopsilosis and Candida metapsilosis spp. nov. to replace Candida parapsilosis groups II and III.
Two new species, Candida orthopsilosis and C. metapsilosis, are proposed to replace the existing designations of C. parapsilosis groups II and III, respectively. The species C. parapsilosis is retained for group I isolates. Attempts to construct a multilocus sequence typing scheme to differentiate individual strains of C. parapsilosis instead revealed fixed DNA sequence differences between pairs of subgroups in four genes: COX3, L1A1, SADH, and SYA1. PCR amplicons for sequencing were obtained for these four plus a further seven genes from 21 group I isolates. For nine group II isolates, PCR products were obtained from only 5 of the 11 genes, and for two group III isolates PCR products were obtained from a different set of 5 genes. Three of the PCR products from group II and III isolates differed in size from the group I products. Cluster analysis of sequence polymorphisms from COX3, SADH, and SYA1, which were common to the three groups, consistently separated the isolates into three distinct sets. All of these differences, together with DNA sequence similarities <90% in the ITS1 sequence, suggest the subgroups should be afforded species status. The near absence of DNA sequence variability among isolates of C. parapsilosis and relatively high levels of sequence variability among isolates of C. orthopsilosis suggest that the former species may have evolved very recently from the latter.


Treat-to-target in vasculitis: is this a sensible approach?
It is possible to achieve substantial initial control of systemic vasculitis in the majority of patients. However, the 'target' has shifted considerably over the last 20-30 years from keeping patients alive to maintaining good quality disease control, avoiding the development of comorbidities - either as a result of disease or treatment, and also preventing relapses. This expansion of potential targets that can be achieved in systemic vasculitis has arisen because we have more effective therapies, but more importantly we have developed a framework within which targets can be created reproducibly. In other words we have much clearer definitions of what constitutes clinical disease activity, relapse, remission and morbidity. These targets are based on simple clinical evaluation, limited laboratory assessments of patients that can be undertaken by any secondary care facility. As a result of this they remain at a clinical level and may not address the most important targets, which are curing disease and that would be the aspiration to move towards. The first step towards that is to move from clinically-based targets towards mechanistic targets based primarily around the pathophysiological drivers of disease. That in turn may lead to identification of specific targets that can turn off disease. The systemic vasculitides are heterogeneous and although for ANCA-associated vasculitis in the short term treatments are similar, the development of clear understanding of mechanisms and new targets may bring with it the promise of much more focused therapies that will address only individual targets and therefore personalize therapy for each individual condition and patient.


Increased Expression of Phosphorylated FADD in Anaplastic Large Cell and Other T-Cell Lymphomas.
FAS-associated protein with death domain (FADD) is a major adaptor protein involved in extrinsic apoptosis, embryogenesis, and lymphocyte homeostasis. Although abnormalities of the FADD/death receptor apoptotic pathways have been established in tumorigenesis, fewer studies have analyzed the expression and role of phosphorylated FADD (pFADD). Our identification of FADD as a lymphoma-associated autoantigen in T-cell lymphoma patients raises the possibility that pFADD, with its correlation with cell cycle, may possess role(s) in human T-cell lymphoma development. This immunohistochemical study investigated pFADD protein expression in a range of normal tissues and lymphomas, particularly T-cell lymphomas that require improved therapies. Whereas pFADD was expressed only in scattered normal T cells, it was detected at high levels in T-cell lymphomas (eg, 84% anaplastic large cell lymphoma and 65% peripheral T cell lymphomas, not otherwise specified). The increased expression of pFADD supports further study of its clinical relevance and role in lymphomagenesis, highlighting phosphorylation of FADD as a potential therapeutic target.


Strategic Trastuzumab Mediated Crosslinking Driving Concomitant HER2 and HER3 Endocytosis and Degradation in Breast Cancer.
Efficacious anticancer therapies for targeting plasma membrane receptors with antibody based therapeutics are often contingent on sufficient endocytic delivery of receptor and conjugate to lysosomes. This results in downregulation of receptor activity and, in the case of antibody-drug conjugates (ADCs), intracellular release of a drug payload. The oncogenic receptor HER2 is a priority therapeutic target in breast cancer. Known as an "endocytosis resistant" receptor, HER2 thwarts the receptor downregulating efficiency of the frontline treatment trastuzumab and reduces the potential of trastuzumab-based therapies such as trastuzumab-emtansine. We previously demonstrated that strategically inducing trastuzumab and HER2 crosslinking in breast cancer cells promoted endocytosis and lysosomal delivery of the HER2-trastuzumab complex, stimulating downregulation of the receptor. Here we reveal that HER3, but not EGFR, is also concomitantly downregulated with HER2 after crosslinking. This is accompanied by strong activation of MEK/ERK pathway that we show does not directly contribute to HER2/trastuzumab endocytosis. We show that crosslinking induced trastuzumab endocytosis occurs via clathrin-dependent and independent pathways and is an actin-dependent process. Detailed ultrastructural studies of the plasma membrane highlight crosslinking-specific remodelling of microvilli and induction of extensive ruffling. Investigations in a cell model of acquired trastuzumab resistance demonstrate, for the first time, that they are refractory to crosslinking induced HER2 endocytosis and downregulation. This implicates further arrest of HER2 internalisation in developing trastuzumab resistance. Overall our findings highlight the potential of receptor crosslinking as a therapeutic strategy for cancer while exposing the ability of cancer cells to develop resistance via endocytic mechanisms.


Combining Vascular Normalization with an Oncolytic Virus Enhances Immunotherapy in a Preclinical Model of Advanced-Stage Ovarian Cancer.
Intravenous delivery of oncolytic viruses often leads to tumor vascular shutdown, resulting in decreased tumor perfusion and elevated tumor hypoxia. We hypothesized that using 3TSR to normalize tumor vasculature prior to administration of an oncolytic Newcastle disease virus (NDV) would enhance virus delivery and trafficking of immunologic cell subsets to the tumor core, resulting in systemically enhanced immunotherapy and regression of advanced-stage epithelial ovarian cancer (EOC). Using an orthotopic, syngeneic mouse model of advanced-stage EOC, we pretreated mice with 3TSR (4 mg/kg per day) alone or followed by combination with fusogenic NDV(F3aa) (1.0 × 108 plaque-forming units). Treatment with 3TSR normalized tumor vasculature, enhanced blood perfusion of primary EOC tumors, and induced disease regression. Animals treated with combination therapy had the greatest reduction in primary tumor mass, ascites accumulation, and secondary lesions (50% of mice were completely devoid of peritoneal metastases). Combining 3TSR + NDV(F3aa) led to enhanced trafficking of immunologic cells into the primary tumor core. We have shown, for the first time, that NDV, like other oncolytic viruses, is a potent mediator of acute vascular shutdown and that preventing this through vascular normalization can promote regression in a preclinical model of advanced-stage ovarian cancer. This challenges the current focus on induction of intravascular thrombosis as a requisite for successful oncolytic virotherapy.See related commentary by Bykov and Zamarin, p. 1446.


Mesenchymal stem cells alleviate airway inflammation and emphysema in COPD through down-regulation of cyclooxygenase-2 via p38 and ERK MAPK pathways.
Bone marrow-derived mesenchymal stem cells (MSCs) have been identified as one possible strategy for the treatment of chronic obstructive pulmonary disease (COPD). Our previous studies have demonstrated that MSC administration has therapeutic potential in airway inflammation and emphysema via a paracrine mechanism. We proposed that MSCs reverse the inflammatory process and restore impaired lung function through their interaction with macrophages. In our study, the rats were exposed to cigarette smoke (CS), followed by the administration of MSCs into the lungs for 5 weeks. Here we show that MSC administration alleviated airway inflammation and emphysema through the down-regulation of cyclooxygenase-2 (COX-2) and COX-2-mediated prostaglandin E2 (PGE2) production, possibly through the effect on alveolar macrophages. In vitro co-culture experiments provided evidence that MSCs down-regulated COX-2/PGE2 in macrophages through inhibition of the activation-associated phosphorylation of p38 MAPK and ERK. Our data suggest that MSCs may relieve airway inflammation and emphysema in CS-exposed rat models, through the inhibition of COX-2/PGE2 in alveolar macrophages, mediated in part by the p38 MAPK and ERK pathways. This study provides a compelling mechanism for MSC treatment in COPD, in addition to its paracrine mechanism.


Studies on desorption and regeneration of natural pumice for iron removal from aqueous solution.
To make the adsorption process more economic and environmental friendly, it is necessary to study desorption and reutilization of the adsorbents. In the present study, the effectiveness of natural pumice in removal of iron from aqueous solution was investigated in several sorption-desorption cycles. The desorption characteristics of previously adsorbed iron ions on natural pumice were tested by various desorbing agents such as HCl, NaOH and aquadest. Among them, HCl showed the highest desorption efficiency (37.89%) with 0.1 M of concentration and 60 min of contact time. The removal efficiency of iron ions in reused natural pumice could be maintained up to 90% in the third cycle of adsorption. The results indicate that although complete desorption was not achieved, natural pumice from Sungai Pasak, West Sumatra, Indonesia, can be sufficiently reused up to three cycles of adsorption-desorption.


Extracellular lipid-mediated signaling in tumor-cell activation and pseudopod protrusion.
We have pioneered an in vitro pseudopod-generation model wherein suspended tumor cells are stimulated to form pseudopods into glass micropipettes in response to soluble collagen type IV (CIV). Pertussis toxin and removing intracellular calcium were found previously to be inhibitory to that process. We now extend those observations to dissect the roles of transmembrane calcium influx and circulating fatty acids on pseudopod extension. Removal of fatty acids from BSA in basal media resulted in abrogation of pseudopod formation, while reconstitution of free fatty acids restored cell pseudopod protrusion. We thus hypothesized that fatty acids may provide necessary pseudopod stimulatory signals. Addition of lysophosphatidic acid (LPA) to the fatty acid-free CIV solution or in an opposite pipette without CIV permitted approximately 50% pseudopod recovery in all pipette directions in a dose-dependent fashion. Thapsigargin (TG), an agent that releases internal calcium stores and causes opening of store-operated calcium channels, restored pseudopod protrusion up to 80% in CIV with fatty acid-free albumin. [Ca(2+)](i) release was non-additive when cells were stimulated by TG and LPA, suggesting overlapping [Ca(2+)](i) stores. The combination of TG and LPA in fatty acid-free albumin fully restored the pseudopod response to CIV. Addition of EGTA to chelate stimulatory media calcium blocked the pseudopod response to CIV in the presence of fatty acids. This indicates that pseudopod protrusion requires transmembrane calcium entry. Thus, extracellular lipids and calcium mobilization are required to complement CIV in pseudopod protrusion from suspended cells.


Effect of muscimol on glucose-stimulated somatostatin and insulin release from the isolated, perfused rat pancreas.
This study examines the effect of muscimol, a high affinity, specific gamma-aminobutyric acid (GABA) agonist, on glucose-stimulated somatostatin and insulin release from the isolated, perfused rat pancreas. Perfusion with low glucose (50 mg/dl) conditions resulted in basal somatostatin release of 46 +/- 4 pg/ml. Basal insulin release was less than 20 microU/ml. High glucose (300 mg/dl) conditions stimulated somatostatin and insulin release. Steady-state levels of somatostatin and insulin release under high glucose conditions were 425 +/- 12 pg/ml and 419 +/- 18 microU/ml, respectively. Perfusion with medium containing 1 microM muscimol inhibited glucose-stimulated somatostatin release by 38%, whereas the course of glucose-stimulated insulin release was unaffected. Tentative conclusions from this study are (1) that GABA is potentially a modulator of islet somatostatin but not insulin release, and (2) the fact that somatostatin, an inhibitor of insulin, can be suppressed 38% without coincidental increase in insulin release seems to indicate that, under high glucose conditions, somatostatin is without a significant paracrine effect on the beta-cells.


Automated volumetric evaluation of stereoscopic disc photography.
To develop a fully automated algorithm (AP) to perform a volumetric measure of the optic disc using conventional stereoscopic optic nerve head (ONH) photographs, and to compare algorithm-produced parameters with manual photogrammetry (MP), scanning laser ophthalmoscope (SLO) and optical coherence tomography (OCT) measurements. One hundred twenty-two stereoscopic optic disc photographs (61 subjects) were analyzed. Disc area, rim area, cup area, cup/disc area ratio, vertical cup/disc ratio, rim volume and cup volume were automatically computed by the algorithm. Latent variable measurement error models were used to assess measurement reproducibility for the four techniques. AP had better reproducibility for disc area and cup volume and worse reproducibility for cup/disc area ratio and vertical cup/disc ratio, when the measurements were compared to the MP, SLO and OCT methods. AP provides a useful technique for an objective quantitative assessment of 3D ONH structures.


HIV-1 transmission amongst men who have sex with men: a probabilistic model incorporating antiretroviral treatment optimism-scepticism, sexual beliefs and sexual behaviour.
To explore the hypotheses that HIV-1 infected patients receiving suppressive antiretroviral therapy believe that they cannot transmit their virus, that this translates to increasing rates of unprotected anal intercourse and new infections, and that sexual practice may be influenced by the HIV-1 serostatus of one's sexual partner. We developed a Bernoulli model for HIV-1 transmission amongst a cross-sectional cohort of 119 HIV-1 positive treated and untreated men having sex with men attending a metropolitan HIV-1 clinic. The model included transmission probabilities for different types of anal intercourse, the local HIV-1 seroprevalence rate, and behavioural data from a validated sexual practice questionnaire and scales of treatment optimism-scepticism and sexual beliefs pertaining to the previous 3 months. Subjects reported partner serostatus as HIV-1 negative or unknown. There were no differences in optimism-scepticism score (p = 0.295) and total sexual belief scores (p = 0.211) according to antiretroviral therapy status. Sex with unknown serostatus partners was higher risk for transmission than sex with HIV-1 negative partners. Most unprotected anal intercourse was practiced by a minority of men, independent of antiretroviral treatment status. Compared to subjects with HIV-1 negative partners, subjects with unknown serostatus partners had more UAI, a 2-fold higher probability of transmitting HIV-1 and were responsible for 2.6 as many new infections. Overall, there were 4 times as many infections predicted to occur amongst unknown serostatus men cf. HIV-1 negative men. A sub-group reported frequent unprotected anal intercourse with their partners. Our model may be used to quantify risk behavior during individual counselling or to develop targeted prevention programmes at the public health level.


Computational approaches to identify promoters and cis-regulatory elements in plant genomes.
The identification of promoters and their regulatory elements is one of the major challenges in bioinformatics and integrates comparative, structural, and functional genomics. Many different approaches have been developed to detect conserved motifs in a set of genes that are either coregulated or orthologous. However, although recent approaches seem promising, in general, unambiguous identification of regulatory elements is not straightforward. The delineation of promoters is even harder, due to its complex nature, and in silico promoter prediction is still in its infancy. Here, we review the different approaches that have been developed for identifying promoters and their regulatory elements. We discuss the detection of cis-acting regulatory elements using word-counting or probabilistic methods (so-called "search by signal" methods) and the delineation of promoters by considering both sequence content and structural features ("search by content" methods). As an example of search by content, we explored in greater detail the association of promoters with CpG islands. However, due to differences in sequence content, the parameters used to detect CpG islands in humans and other vertebrates cannot be used for plants. Therefore, a preliminary attempt was made to define parameters that could possibly define CpG and CpNpG islands in Arabidopsis, by exploring the compositional landscape around the transcriptional start site. To this end, a data set of more than 5,000 gene sequences was built, including the promoter region, the 5'-untranslated region, and the first introns and coding exons. Preliminary analysis shows that promoter location based on the detection of potential CpG/CpNpG islands in the Arabidopsis genome is not straightforward. Nevertheless, because the landscape of CpG/CpNpG islands differs considerably between promoters and introns on the one side and exons (whether coding or not) on the other, more sophisticated approaches can probably be developed for the successful detection of "putative" CpG and CpNpG islands in plants.


Unsupervised embedding of single-cell Hi-C data.
Single-cell Hi-C (scHi-C) data promises to enable scientists to interrogate the 3D architecture of DNA in the nucleus of the cell, studying how this structure varies stochastically or along developmental or cell-cycle axes. However, Hi-C data analysis requires methods that take into account the unique characteristics of this type of data. In this work, we explore whether methods that have been developed previously for the analysis of bulk Hi-C data can be applied to scHi-C data. We apply methods designed for analysis of bulk Hi-C data to scHi-C data in conjunction with unsupervised embedding. We find that one of these methods, HiCRep, when used in conjunction with multidimensional scaling (MDS), strongly outperforms three other methods, including a technique that has been used previously for scHi-C analysis. We also provide evidence that the HiCRep/MDS method is robust to extremely low per-cell sequencing depth, that this robustness is improved even further when high-coverage and low-coverage cells are projected together, and that the method can be used to jointly embed cells from multiple published datasets.


Spatial and annual temporal distribution of ozone concentrations in the madrid basin using passive samplers.
Passive samplers are useful tools for helping to describe the ozone distribution in complex terrain situations. They are also a good complement to continuous monitoring stations. This paper discusses the results of a pilot study that used ozone passive samplers to describe the spatial and annual temporal distribution of ozone in several forested areas around the city of Madrid. The ozone concentrations around Madrid were found to be higher on the elevated sites located at a certain distance from the city"s urban zone. A seasonal ozone cycle was observed, with maximum concentrations found in the basin in late spring or summer depending on the location. The information obtained allowed us to group the locations into four classes. Altitude and distance to the city during the summer and winter explained the observed ozone concentrations. However, during the transition periods, especially in early spring and to a lesser extent in autumn, there was not a good correlation between ozone levels and elevation or distance from precursor sources. These data strongly suggest that altitudinal gradients for ozone are not always the case in the Madrid Basin.


Endovascular correction of cerebrovenous anomalies in multiple sclerosis: a retrospective review of an uncontrolled case series.
Endovascular intervention for obstruction to venous drainage of the head and neck is an established treatment for disorders such as superior vena cava syndrome. Some patients with multiple sclerosis have been observed to have anomalies of the veins draining the head and neck. It is possible that some symptoms associated with multiple sclerosis may be secondary to disturbed venous flow. In an uncontrolled clinical series of 40 patients who had been previously diagnosed with multiple sclerosis, anomalies of the venous drainage of the head and neck were observed, including venous stenoses of the internal jugular veins. In 38 of 40 patients, venous stents were placed with restoration of luminal dimensions and abrogation of the venous pressure gradient. The angiographic and hemodynamic improvement was associated with improvement in symptomatology, most particularly in cognitive and constitutional symptoms that may be related to cerebrovenous flow. Serious complications included death in one subject and stent embolization requiring open heart surgery in another. In conclusion, in this series, endovascular intervention to correct venous stenosis associated with multiple sclerosis was associated with improvement in symptoms possibly related to disturbed venous hemodynamics. However, given the serious adverse events in this small series, a randomized clinical trial is required to confirm these findings, and to determine if the procedure has any effect on the progression of multiple sclerosis, or untoward long-term adverse effects.


Ganglioside GD2 identifies breast cancer stem cells and promotes tumorigenesis.
Cancer stem cells (CSCs) are a small subpopulation of cancer cells that have increased resistance to conventional therapies and are capable of establishing metastasis. However, only a few biomarkers of CSCs have been identified. Here, we report that ganglioside GD2 (a glycosphingolipid) identifies a small fraction of cells in human breast cancer cell lines and patient samples that are capable of forming mammospheres and initiating tumors with as few as 10 GD2+ cells. In addition, the majority of GD2+ cells are also CD44hiCD24lo, the previously established CSC-associated cell surface phenotype. Gene expression analysis revealed that GD3 synthase (GD3S) is highly expressed in GD2+ as well as in CD44hiCD24lo cells and that interference with GD3S expression, either by shRNA or using a pharmacological inhibitor, reduced the CSC population and CSC-associated properties. GD3S knockdown completely abrogated tumor formation in vivo. Also, induction of epithelial-mesenchymal transition (EMT) in transformed human mammary epithelial cells (HMLER cells) dramatically increased GD2 as well as GD3S expression in these cells, suggesting a role of EMT in the origin of GD2+ breast CSCs. In summary, we identified GD2 as a new CSC-specific cell surface marker and GD3S as a potential therapeutic target for CSCs, with the possibility of improving survival and cure rates in patients with breast cancer.


Cardioprotective mechanisms of lifestyle modifications and pharmacotherapies on cardiac remodeling and dysfunction in hypertensive heart disease: an overview.
The incidence of cardiovascular events in hypertensive patients is clearly related to a left ventricular mass during treatment, and a regression of left ventricular hypertrophy is associated with a better prognosis. This is the case even independently of changes in other risk factors, including blood pressure. Evidence indicates that lifestyle modifications such as dietary salt restriction and weight loss are effective means in preventing the development of hypertension and reducing blood pressure and left ventricular mass in hypertensive patients. Salt restriction may also reduce the long-term risk of cardiovascular events. It has been recognized that the primary targets of current antihypertensive drugs are the renin-angiotensin-aldosterone system, calcium homeostasis, the ionic transport mechanisms in the kidneys, and the sympathetic nervous system. Clinical as well as experimental studies have demonstrated the cardioprotective effects of antihypertensive drugs independently of their blood pressure lowering effects. Hypertension is often complicated by other disease states including diabetes, dyslipidemia, and ischemic heart disease. Some of the drugs used for the treatment of such complications are also shown to produce cardioprotective effects in addition to their original effects. We ought to better understand these pleiotropic effects for the most effective treatments of hypertension and its complications.


The role of leukocytes in the production of oxygen-derived free radicals in acute experimental pancreatitis.
Oxygen-derived free radicals mediate an important step in the initiation of experimental acute pancreatitis in the ex vivo perfused canine pancreas model. In other organ systems, circulating leukocytes may serve as one source of oxygen-derived free radical production. The current experiments were designed to evaluate the role of circulating leukocytes in the generation of injury in this model. Four experimental groups of animals were studied: group I consisted of controls (n = 6); group II had white blood cell (WBC) depletion (n = 4) in which the recirculating whole blood perfusate was depleted of 98% of its circulating leukocytes; group III had oleic acid infusion (FFA) alone (n = 9), which induced pancreatitis; group IV had WBC depletion and FFA (n = 6), in which oleic acid was infused after depletion of the circulating leukocytes in the perfusate. During the 4-hour perfusion period, the pancreatic preparations were monitored hourly for the development of edema, weight gain, and release of alpha-amylase into the perfusate. Animals in groups I and group II manifested no gross edema, gained minimal weight, and did not manifest hyperamylasemia. Leukocyte depletion alone had no effect. In group IV animals marked edema, significant weight gain, and hyperamylasemia developed to the same extent as in group III animals. These results demonstrate that circulating leukocytes are not essential to the development of pancreatitis in this model and suggest that another source of oxygen-derived free radicals mediates this injury.


Effects of Bothrops asper snake venom on lymphatic vessels: insights into a hidden aspect of envenomation.
Envenomations by the snake Bothrops asper represent a serious medical problem in Central America and parts of South America. These envenomations concur with drastic local tissue pathology, including a prominent edema. Since lymph flow plays a role in the maintenance of tissue fluid balance, the effect of B. asper venom on collecting lymphatic vessels was studied. B. asper venom was applied to mouse mesentery, and the effects were studied using an intravital microscopy methodology coupled with an image analysis program. B. asper venom induced a dose-dependent contraction of collecting lymphatic vessels, resulting in a reduction of their lumen and in a halting of lymph flow. The effect was reproduced by a myotoxic phospholipase A(2) (PLA(2)) homologue isolated from this venom, but not by a hemorrhagic metalloproteinase or a coagulant thrombin-like serine proteinase. In agreement with this, treatment of the venom with fucoidan, a myotoxin inhibitor, abrogated the effect, whereas no inhibition was observed after incubation with the peptidomimetic metalloproteinase inhibitor Batimastat. Moreover, fucoidan significantly reduced venom-induced footpad edema. The myotoxic PLA(2) homologue, known to induce skeletal muscle necrosis, was able to induce cytotoxicity in smooth muscle cells in culture and to promote an increment in the permeability to propidium iodide in these cells. Our observations indicate that B. asper venom affects collecting lymphatic vessels through the action of myotoxic PLA(2)s on the smooth muscle of these vessels, inducing cell contraction and irreversible cell damage. This activity may play an important role in the pathogenesis of the pronounced local edema characteristic of viperid snakebite envenomation, as well as in the systemic biodistribution of the venom, thus representing a potential therapeutical target in these envenomations.


Performance of the integrated management of childhood illness algorithm for diagnosis of HIV-1 infection among African infants.
Early infant HIV-1 diagnosis and treatment substantially improve survival. Where virologic HIV-1 testing is unavailable, integrated management of childhood illness (IMCI) clinical algorithms may be used for infant HIV-1 screening. We evaluated the performance of the 2008 WHO IMCI HIV algorithm in a cohort of HIV-exposed Kenyan infants. From 1999 to 2003, 444 infants had monthly clinical assessments and quarterly virologic HIV-1 testing. Using archived clinical data, IMCI sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were calculated using virologic testing as a gold standard. Linear regression and survival analyses were used to determine the effect of age on IMCI performance and timing of diagnosis. Overall IMCI sensitivity, specificity, PPV, and NPV value were 58, 87, 52, and 90%, respectively. Sensitivity (1.4%) and PPV (14%) were lowest at 1 month of age, when 81% of HIV infections already had occurred. Sensitivity increased with age (P < 0.0001), but remained low throughout infancy (range 1.4-35%). Specificity (range 97-100%) was high at each time point and was not associated with age. Fifty-eight percent of HIV-1-infected infants (50 of 86) were eventually diagnosed by IMCI, and use of IMCI was estimated to delay diagnosis in HIV-infected infants by a median of 5.9 months (P < 0.0001). IMCI had low sensitivity during the first month of life, when the majority of HIV-1 infections had already occurred and initiation of treatment is most critical. Although sensitivity increased with age, the substantial delay in HIV-1 diagnosis using IMCI limits its utility in early infant HIV-1 diagnosis.


An asymptotically consistent approximant method with application to soft- and hard-sphere fluids.
A modified Padé approximant is used to construct an equation of state, which has the same large-density asymptotic behavior as the model fluid being described, while still retaining the low-density behavior of the virial equation of state (virial series). Within this framework, all sequences of rational functions that are analytic in the physical domain converge to the correct behavior at the same rate, eliminating the ambiguity of choosing the correct form of Padé approximant. The method is applied to fluids composed of "soft" spherical particles with separation distance r interacting through an inverse-power pair potential, φ = ε(σ∕r)(n), where ε and σ are model parameters and n is the "hardness" of the spheres. For n < 9, the approximants provide a significant improvement over the 8-term virial series, when compared against molecular simulation data. For n ≥ 9, both the approximants and the 8-term virial series give an accurate description of the fluid behavior, when compared with simulation data. When taking the limit as n → ∞, an equation of state for hard spheres is obtained, which is closer to simulation data than the 10-term virial series for hard spheres, and is comparable in accuracy to other recently proposed equations of state. By applying a least square fit to the approximants, we obtain a general and accurate soft-sphere equation of state as a function of n, valid over the full range of density in the fluid phase.


Free energy calculations and molecular properties of substrate translocation through OccAB porins.
One of the greatest health threats facing modern medicine is the emergence of new bacterial strains which are increasingly resistant to almost all currently available antibiotics. According to a CDC (Center for Disease Control and Prevention) report published in 2013, 63% of Acinetobacter species have been identified as Multidrug resistant strains. As for other Gram-negative bacteria, the presence of an outer membrane increases the intrinsic resistance of A. baumannii to most antibiotics. The outer membrane of A. baumannii possesses several specific porins that control the selectivity for different polar substrates in a way that is still poorly understood. Recently, the X-ray crystal structures of 4 related porins, termed OccAB1-4, were solved at high resolution, providing a framework to study the structural and functional characteristics of these porins in filtering natural substrates. Here, we first use molecular dynamics simulations on OccAB proteins to investigate the stability and dynamics of the pores, and to establish their common biophysical features. We then applied metadynamics simulations to evaluate the free energy costs required for polar substrates to overcome the pore. Together, the comparative analysis of the OccAB porins not only sheds light on how these channels could function as potential antibiotic gateways, but also allows identification of putative affinity sites that represent a common path through which other molecules can transit.


Rates of Perioperative Complications Among Patients Undergoing Orthopedic Trauma Surgery Despite Having Positive Results for Methamphetamine.
The burden of psychosocial problems, including substance abuse, is high among trauma patients. Use of illicit substances is often noted during urine toxicology screening on admission and can delay surgery because of concerns for an interaction with anesthesia. Methamphetamine theoretically has potential to increase perioperative anesthetic risks. However, the authors are unaware of any studies documenting increased rates of cardiovascular complications in the perioperative period among orthopedic trauma patients. This study sought to determine the rate of cardiovascular complications in these patients. The authors reviewed the medical records of all patients between 2013 and 2018 who underwent orthopedic trauma surgery at two level I trauma centers in the setting of a methamphetamine-positive urine toxicology screening prior to surgery. Information on demographics, injury, type of surgical intervention, and incidence of perioperative cardiovascular and overall medical complications prior to discharge was recorded. Ninety-four patients were included in the study (mean age, 44 years; range, 16-78 years). Twenty-six (28%) patients had multiple injuries. Thirteen (14%) patients had debridement and/or provisional stabilization of an open or unstable fracture, 18 (19%) had treatment for an infection, and 63 (67%) had definitive fracture surgery. The overall rates of perioperative cardiovascular complications and perioperative medical complications were 2.1% and 3.2%, respectively. This study provides both a baseline understanding of the complication rate for methamphetamine-positive orthopedic trauma patients during general anesthesia and justification for larger multicenter studies to further investigate this topic. [Orthopedics. 2019; 42(4):192-196.].


Antibiotics and antiseptics for venous leg ulcers.
Venous leg ulcers are a type of chronic wound affecting up to 1% of adults in developed countries at some point during their lives. Many of these wounds are colonised by bacteria or show signs of clinical infection. The presence of infection may delay ulcer healing. Two main strategies are used to prevent and treat clinical infection in venous leg ulcers: systemic antibiotics and topical antibiotics or antiseptics. The objective of this review was to determine the effects of systemic antibiotics and topical antibiotics and antiseptics on the healing of venous ulcers. In May 2013, for this second update, we searched the Cochrane Wounds Group Specialised Register (searched 24 May 2013); the Cochrane Central Register of Controlled Trials (CENTRAL 2013, Issue 4); Ovid MEDLINE (1948 to Week 3 May 2013); Ovid MEDLINE (In-Process & Other Non-indexed Citations, 22 May 2013); Ovid EMBASE (1980 to Week 20 2013); and EBSCO CINAHL (1982 to 17 May 2013). No language or publication date restrictions were applied. Randomised controlled trials (RCTs) recruiting people with venous leg ulceration, evaluating at least one systemic antibiotic, topical antibiotic or topical antiseptic that reported an objective assessment of wound healing (e.g. time to complete healing, frequency of complete healing, change in ulcer surface area) were eligible for inclusion. Selection decisions were made by two review authors while working independently. Information on the characteristics of participants, interventions and outcomes was recorded on a standardised data extraction form. In addition, aspects of trial methods were extracted, including randomisation, allocation concealment, blinding of participants and outcome assessors, incomplete outcome data and study group comparability at baseline. Data extraction and validity assessment were conducted by one review author and were checked by a second. Data were pooled when appropriate. Forty-five RCTs reporting 53 comparisons and recruiting a total of 4486 participants were included, Many RCTs were small, and most were at high or unclear risk of bias. Ulcer infection status at baseline and duration of follow-up varied across RCTs. Five RCTs reported eight comparisons of systemic antibiotics, and the remainder evaluated topical preparations: cadexomer iodine (11 RCTs reporting 12 comparisons); povidone-iodine (six RCTs reporting seven comparisons); peroxide-based preparations (four RCTs reporting four comparisons); honey-based preparations (two RCTs reporting two comparisons); silver-based preparations (12 RCTs reporting 13 comparisons); other topical antibiotics (three RCTs reporting five comparisons); and other topical antiseptics (two RCTs reporting two comparisons). Few RCTs provided a reliable estimate of time to healing; most reported the proportion of participants with complete healing during the trial period. Systemic antibioticsMore participants were healed when they were prescribed levamisole (normally used to treat roundworm infection) compared with placebo: risk ratio (RR) 1.31 (95% CI 1.06 to 1.62). No between-group differences were detected in terms of complete healing for other comparisons: antibiotics given according to antibiogram versus usual care; ciprofloxacin versus standard care/placebo; trimethoprim versus placebo; ciprofloxacin versus trimethoprim; and amoxicillin versus topical povidone-iodine. Topical antibiotics and antiseptics Cadexomer iodine: more participants were healed when given cadexomer iodine compared with standard care. The pooled estimate from four RCTs for complete healing at four to 12 weeks was RR 2.17 (95% CI 1.30 to 3.60). No between-group differences in complete healing were detected when cadexomer iodine was compared with the following: hydrocolloid dressing; paraffin gauze dressing; dextranomer; and silver-impregnated dressings.Povidone iodine: no between-group differences in complete healing were detected when povidone-iodine was compared with the following: hydrocolloid; moist or foam dressings according to wound status; and growth factor. Time to healing estimates for povidone-iodine versus dextranomer, and for povidone-iodine versus hydrocolloid, were likely to be unreliable.Peroxide-based preparations: four RCTs reported findings in favour of peroxide-based preparations when compared with usual care for surrogate healing outcomes (change in ulcer area). There was no report of complete healing.Honey-based preparations: no between-group difference in time to healing or complete healing was detected for honey-based products when compared with usual care.Silver-based preparations: no between-group differences in complete healing were detected when 1% silver sulphadiazine ointment was compared with standard care/placebo and tripeptide copper complex; or when different brands of silver-impregnated dressings were compared; or when silver-impregnated dressings were compared with non-antimicrobial dressings.Other topical antibiotics: data from one RCT suggested that more participants healed at four weeks when treated with an enzymatic cleanser (a non-antibiotic preparation) compared with a chloramphenicol-containing ointment (additional active ingredients also included in the ointment): RR 0.13 (95% CI 0.02 to 0.99). No between-group differences in complete healing were detected for framycetin sulphate ointment versus enzymatic cleanser; chloramphenicol ointment versus framycetin sulphate ointment; mupirocin ointment versus vehicle; and topical antibiotics given according to antibiogram versus an herbal ointment.Other topical antiseptics: data from one RCT suggested that more participants receiving an antiseptic ointment (ethacridine lactate) had responsive ulcers (defined as > 20% reduction in area) at four weeks when compared with placebo: RR 1.45 (95% CI 1.21 to 1.73). Complete healing was not reported. No between-group difference was detected between chlorhexidine solution and usual care. At present, no evidence is available to support the routine use of systemic antibiotics in promoting healing of venous leg ulcers. However, the lack of reliable evidence means that it is not possible to recommend the discontinuation of any of the agents reviewed. In terms of topical preparations, some evidence supports the use of cadexomer iodine. Current evidence does not support the routine use of honey- or silver-based products. Further good quality research is required before definitive conclusions can be drawn about the effectiveness of povidone-iodine, peroxide-based preparations, ethacridine lactate, chloramphenicol, framycetin, mupirocin, ethacridine or chlorhexidine in healing venous leg ulceration. In light of the increasing problem of bacterial resistance to antibiotics, current prescribing guidelines recommend that antibacterial preparations should be used only in cases of clinical infection, not for bacterial colonisation.


"I can make more from selling medicine when breaking the rules" - understanding the antibiotic supply network in a rural community in Viet Nam.
As in many other low and middle income countries (LIMCs), Vietnam has experienced a major growth in the pharmaceutical industry, with large numbers of pharmacies and drug stores, and increasing drug expenditure per capita over the past decade. Despite regulatory frameworks that have been introduced to control the dispensing and use of prescription-only drugs, including antibiotics, compliance has been reported to be strikingly low particularly in rural parts of Vietnam. This qualitative study aimed to understand antibiotic access and use practices in the community from both supplier and consumer perspectives in order to support the identification and development of future interventions. This qualitative study was part of a project on community antibiotic access and use (ABACUS) in six LMICs. The focus was Ba Vi district of Hanoi capital city, where we conducted 16 indepth interviews (IDIs) with drug suppliers, and 16 IDIs and 6 focus group discussions (FGDs) with community members. Drug suppliers were sampled based on mapping of all informal and formal antibiotic purchase or dispensing points in the study area. Community members were identified through local networks and relationships with the field collaborators. All IDIs and FGDs were audio-taped, transcribed and analysed using content analysis. We identified a large number of antibiotic suppliers in the locality with widespread infringements of regulatory requirements. Established reciprocal relationships between suppliers and consumers in drug transactions were noted, as was the consumers' trust in the knowledge and services provided by the suppliers. In addition, antibiotic use has become a habitual choice in most illness conditions, driven by both suppliers and consumers. This study presents an analysis of the practices of antibiotic access and use in a rural Vietnamese setting. It highlights the interactions between antibiotic suppliers and consumers in the community and identifies possible targets for interventions.


Recyclable (Fe3O4-NaYF4:Yb,Tm)@TiO2 nanocomposites with near-infrared enhanced photocatalytic activity.
Herein, the design and synthesis of a multifunctional (Fe3O4-NaYF4:Yb,Tm)@TiO2 photocatalyst through a facile sol-gel process combined with electrostatic self-assembly has been reported. Particularly, as an upconversion sensor, NaYF4:Yb,Tm nanoparticles have the ability to convert near-infrared (NIR) light into ultraviolet (UV) emissions, so that the catalytic body TiO2 can indirectly utilize the NIR part of sunlight but not UV light. The excellent photocatalytic activity of the hybrid samples is evaluated using a model of the degradation of methylene blue (MB). Most importantly, the obtained materials exhibit remarkable magnetic properties because of the addition of a magnetic component (Fe3O4), therefore, the photocatalysts possess desired recyclability and reusability, which is significant for actual applications. Finally, the possible photocatalysis mechanism of the nanohybrids is discussed and hydroxyl radicals (OH˙) are confirmed as the main reactive species.


Efficacy of Mycobacterium indicus pranii immunotherapy as an adjunct to chemotherapy for tuberculosis and underlying immune responses in the lung.
The 9-month-long chemotherapy of tuberculosis often results in poor compliance and emergence of drug-resistant strains. So, improved therapeutic strategy is urgently needed. Immunotherapy could be beneficial for the effective management of the disease. Previously we showed the protective efficacy of Mycobacterium indicus pranii (MIP) when given as prophylactic vaccine in animal models of tuberculosis. We sought to investigate whether MIP can be used as an adjunct to the chemotherapy in guinea pig models of tuberculosis. Efficacy of MIP was evaluated when given subcutaneously or by aerosol. MIP-therapy as an adjunct to the chemotherapy was found to be effective in accelerating bacterial killing and improving organ pathology. MIP-immunotherapy resulted in higher numbers of activated antigen-presenting cells and lymphocytes in the infected lungs and also modulated the granulomatous response. Early increase in protective Th1 immune response was observed in the immunotherapy group. Following subsequent doses of MIP, decrease in the inflammatory response and increase in the immunosuppressive response was observed, which resulted in the improvement of lung pathology. MIP immunotherapy is a valuable adjunct to chemotherapy for tuberculosis. Aerosol route of immunotherapy can play a crucial role for inducing immediate local immune response in the lung.


CECR2, a protein involved in neurulation, forms a novel chromatin remodeling complex with SNF2L.
Chromatin remodeling complexes play critical roles in development. Here we describe a transcription factor, CECR2, which is involved in neurulation and chromatin remodeling. CECR2 shows complex alternative splicing, but all variants contain DDT and bromodomain motifs. A mutant mouse line was generated from an embryonic stem cell line containing a genetrap within Cecr2. Reporter gene expression demonstrated Cecr2 expression to be predominantly neural in the embryo. Mice homozygous for the Cecr2 genetrap-induced mutation show a high penetrance of the neural tube defect exencephaly, the human equivalent of anencephaly, in a strain-dependent fashion. Biochemical isolation of CECR2 revealed the presence of this protein as a component of a novel heterodimeric complex termed CECR2-containing remodeling factor (CERF). CERF comprises CECR2 and the ATP-dependent chromatin remodeler SNF2L, a mammalian ISWI ortholog expressed predominantly in the central nervous system. CERF is capable of remodeling chromatin in vitro and displays an ATP hydrolyzing activity that is stimulated by nucleosomes. Together, these data identify a novel chromatin remodeling complex with a critical role in neurulation.


Characteristic Sizes of Life in the Oceans, from Bacteria to Whales.
The size of an individual organism is a key trait to characterize its physiology and feeding ecology. Size-based scaling laws may have a limited size range of validity or undergo a transition from one scaling exponent to another at some characteristic size. We collate and review data on size-based scaling laws for resource acquisition, mobility, sensory range, and progeny size for all pelagic marine life, from bacteria to whales. Further, we review and develop simple theoretical arguments for observed scaling laws and the characteristic sizes of a change or breakdown of power laws. We divide life in the ocean into seven major realms based on trophic strategy, physiology, and life history strategy. Such a categorization represents a move away from a taxonomically oriented description toward a trait-based description of life in the oceans. Finally, we discuss life forms that transgress the simple size-based rules and identify unanswered questions.


Imaging diagnosis-magnetic resonance imaging features of metastatic cerebral lymphoma in a dog.
We describe histopathologically confirmed intracranial metastasis of cutaneous lymphoma. In magnetic resonance (MR) images there was a heterogeneous, contrast-enhancing, extraaxial mass in the right parietal and piriform lobes at the level of the optic chiasm. Our MR imaging findings are consistent with reports in humans in that lymphoma masses have indistinct borders that are iso- to hyperintense relative to adjacent gray matter on T2-weighted images. Our report varies from findings in humans in that the mass was extraaxial, whereas masses reported in humans are intraaxial. Contrast enhancement can be heterogeneous, as in our report, or homogeneous.


An integrated risk and vulnerability assessment framework for climate change and malaria transmission in East Africa.
Malaria is one of the key research concerns in climate change-health relationships. Numerous risk assessments and modelling studies provide evidence that the transmission range of malaria will expand with rising temperatures, adversely impacting on vulnerable communities in the East African highlands. While there exist multiple lines of evidence for the influence of climate change on malaria transmission, there is insufficient understanding of the complex and interdependent factors that determine the risk and vulnerability of human populations at the community level. Moreover, existing studies have had limited focus on the nature of the impacts on vulnerable communities or how well they are prepared to cope. In order to address these gaps, a systems approach was used to present an integrated risk and vulnerability assessment framework for studies of community level risk and vulnerability to malaria due to climate change. Drawing upon published literature on existing frameworks, a systems approach was applied to characterize the factors influencing the interactions between climate change and malaria transmission. This involved structural analysis to determine influential, relay, dependent and autonomous variables in order to construct a detailed causal loop conceptual model that illustrates the relationships among key variables. An integrated assessment framework that considers indicators of both biophysical and social vulnerability was proposed based on the conceptual model. A major conclusion was that this integrated assessment framework can be implemented using Bayesian Belief Networks, and applied at a community level using both quantitative and qualitative methods with stakeholder engagement. The approach enables a robust assessment of community level risk and vulnerability to malaria, along with contextually relevant and targeted adaptation strategies for dealing with malaria transmission that incorporate both scientific and community perspectives.


Formation and origin of basal lamina and anchoring fibrils in adult human skin.
The purpose of this investigation was to study the formation and origin of basal lamina and anchoring fibrils in adult human skin. Epidermis and dermis were separated by "cold trypsinization." Viable epidermis and viable, inverted dermis were recombined and grafted to the chorioallantoic membrane of embryonated chicken eggs for varying periods up to 10 days. Basal lamina and anchoring fibrils were absent from the freshly trypsinized epidermis before grafting although hemidesmosomes and tonofilaments of the basal cells remained intact. Basal lamina and anchoring fibrils were absent from freshly cut, inverted surface of the dermis. Beginning 3 days after grafting, basal lamina was noted to form immediately subjacent to hemidesmosomes of epidermal basal cells at the epidermal-dermal interface. From the fifth to the seventh day after grafting, basal lamina became progressively more dense and extended to become continuous in many areas at the epidermal-dermal interface. Anchoring fibrils appeared first in grafts consisting of epidermis and viable dermis at five day cultivation and became progressively more numerous thereafter. In order to determine the epidermal versus dermal origin of basal lamina and anchoring fibrils, dermis was rendered nonviable by repeated freezing and thawing 10 times followed by recombination with viable epidermis. Formation of basal lamina occurred as readily in these recombinants of epidermis with freeze-thawed, nonviable dermis as with viable dermis, indicating that dermal viability was not essential for synthesis of basal lamina. This observation supports the concept of epidermal origin for basal lamina. Anchoring fibrils did not form in recombinants containing freeze-thawed dermis, indicating that dermal viability was required for anchoring fibrils formation. This observation supports the concept of dermal origin of anchoring fibrils.


What's going on with young people today? the long and twisting path to adulthood.
Richard Settersten and Barbara Ray examine the lengthening transition to adulthood over the past several decades, as well as the challenges the new schedule poses for young people, families, and society. The authors begin with a brief history of becoming an adult, noting that the schedule that youth follow to arrive at adulthood changes to meet the social realities of each era. For youth to leave home at an early age during the 1950s, for example, was "normal" because opportunities for work were plentiful and social expectations of the time reinforced the need to do so. But the prosperity that made it possible for young adults of that era to move quickly into adult roles did not last. The economic and employment uncertainties that arose during the 1970s complicated enormously the decisions that young adults had to make about living arrangements, educational investments, and family formation. The authors next take a closer look at changes in the core timing shifts in the new transition-the lengthening time it now takes youth to leave home, complete school, enter the workforce, marry, and have children. They stress that today's new schedule for attaining independence leaves many families overburdened as they support their children for an extended period. The continued need to rely on families for financial assistance, the authors say, exacerbates the plight of young people from a variety of vulnerable backgrounds. It also raises complex questions about who is responsible for the welfare of young people and whether the risks and costs newly associated with the early adult years should be absorbed by markets, by families, or by governments. Settersten and Ray stress that the longer transition to adulthood strains not only families but also the institutions that have traditionally supported young Americans in making that transition-such as residential colleges and universities, community colleges, military service, and national service programs. They emphasize the need to strengthen existing social institutions and create new ones to reflect more accurately the realities of a longer and more complex passage into adult life.


Seasonal variations of selected cardiovascular risk factors.
This article reviews research on selected biomarkers of cardiovascular risk - cholesterol and other lipids, C-reactive protein (CRP), fibrinogen, homocysteine - in the attempt to determine the existence of a predictable seasonal chronobiological pattern of variation. Studies dating as far back as the 1930s have reported seasonal variations in cholesterol levels. Statistically significant seasonal changes in lipid levels have been found in individuals irrespective of the country where the research has been conducted, and irrespective of the age, sex, ethnicity, and baseline lipid levels of the study subjects. While not all studies have been in complete agreement on either the amplitude (degree of seasonal change) or month/s of highest lipid levels, a strong winter/summer difference has been found in most studies. Existing evidence for an independent effect of season in variation of CRP is weak. Studies have consistently reported significant seasonal variations in fibrinogen levels. While other biological factors clearly interact to affect fibrinogen variability, seasonality appears to be an independent source of variability. Evidence from several studies points to a lack of seasonal variability in homocysteine levels. Although seasonal variability is just one source of periodicity influencing biological function and assessments in clinical practice, for some biomarkers, including lipids and fibrinogen, it is a source of variability that warrants consideration prior to a decision to treat and in assessing response to interventions.


Role of nitric oxide deficiency in the development of hypertension in hydronephrotic animals.
Hydronephrotic animals develop renal injury and hypertension, which is associated with an abnormal tubuloglomerular feedback (TGF). The TGF sensitivity is coupled to nitric oxide (NO) in the macula densa. The involvement of reduced NO availability in the development of hypertension in hydronephrosis was investigated. Hydronephrosis was induced by ureteral obstruction in young rats. Blood pressure and renal excretion were measured in adulthood, under different sodium conditions, and before and after chronic administration of either N(G)-nitro-l-arginine methyl ester (l-NAME) or l-arginine. Blood samples for ADMA, SDMA, and l-arginine analysis were taken and the renal tissue was used for histology and determination of NO synthase (NOS) proteins. TGF characteristics were determined by stop-flow pressure technique before and after administration of 7-nitroindazole (7-NI) or l-arginine. Hydronephrotic animals developed salt-sensitive hypertension, which was associated with pressure natriuresis and diuresis. The blood pressure response to l-NAME was attenuated and l-arginine supplementation decreased blood pressure in hydronephrotic animals, but not in the controls. Under control conditions, reactivity and sensitivity of the TGF response were greater in the hydronephrotic group. 7-NI administration increased TGF reactivity and sensitivity in control animals, whereas, in hydronephrotic animals, neuronal NOS (nNOS) inhibition had no effect. l-Arginine attenuated TGF response more in hydronephrotic kidneys than in controls. The hydronephrotic animals displayed various degrees of histopathological changes. ADMA and SDMA levels were higher and the renal expressions of nNOS and endothelial NOS proteins were lower in animals with hydronephrosis. Reduced NO availability in the diseased kidney in hydronephrosis, and subsequent resetting of the TGF mechanism, plays an important role in the development of hypertension.


Forty-Year Trends in Tooth Loss Among American Adults With and Without Diabetes Mellitus: An Age-Period-Cohort Analysis.
This study aimed to assess the trends in tooth loss among adults with and without diabetes mellitus in the United States and racial/ethnic disparities in tooth loss patterns, and to evaluate trends in tooth loss by age, birth cohorts, and survey periods. Data came from 9 waves of the National Health and Nutrition Examination Survey (NHANES) from 1971 through 2012. The trends in the estimated tooth loss in people with and without diabetes were assessed by age groups, survey periods, and birth cohorts. The analytical sample was 37,609 dentate (ie, with at least 1 permanent tooth) adults aged 25 years or older. We applied hierarchical age-period-cohort cross-classified random-effects models for the trend analysis. The estimated number of teeth lost among non-Hispanic blacks with diabetes increased more with age than that among non-Hispanic whites with diabetes (z = 4.05, P < .001) or Mexican Americans with diabetes (z = 4.38, P < .001). During 1971-2012, there was a significant decreasing trend in the number of teeth lost among non-Hispanic whites with diabetes (slope = -0.20, P < .001) and non-Hispanic blacks with diabetes (slope = -0.37, P < .001). However, adults with diabetes had about twice the tooth loss as did those without diabetes. Substantial differences in tooth loss between adults with and without diabetes and across racial/ethnic groups persisted over time. Appropriate dental care and tooth retention need to be further promoted among adults with diabetes.


A Comparison of Microdensitometry and TV Waveform Analysis as Expressions of Observed Landscape Patterns on Radar.
Delimiting landscape regions with radar is largely dependent upon recognition of variations in texture patterns on the radar image. Since texture is a function of frequency and magnitude of tone or density changes over discrete distances any objective means of evaluating textures must be based on mechanically measuring and recording tonal information. Comparison of microdensitometer traces with television waveforms reveals that, for similar resolution systems, essentially the same textural information can be obtained from either system. Many interpreters who do not have access to scanning densitometers do have access to television equipment. This, and the fact that the location of the scan line being analyzed can be viewed simultaneously on the television monitor, favors waveform analysis for many interpretation situations.


Dose-dependent social-cognitive effects of intranasal oxytocin delivered with novel Breath Powered device in adults with autism spectrum disorder: a randomized placebo-controlled double-blind crossover trial.
The neuropeptide oxytocin has shown promise as a treatment for symptoms of autism spectrum disorders (ASD). However, clinical research progress has been hampered by a poor understanding of oxytocin's dose-response and sub-optimal intranasal delivery methods. We examined two doses of oxytocin delivered using a novel Breath Powered intranasal delivery device designed to improve direct nose-to-brain activity in a double-blind, crossover, randomized, placebo-controlled trial. In a randomized sequence of single-dose sessions, 17 male adults with ASD received 8 international units (IU) oxytocin, 24IU oxytocin or placebo followed by four social-cognitive tasks. We observed an omnibus main effect of treatment on the primary outcome measure of overt emotion salience as measured by emotional ratings of faces (η2=0.18). Compared to placebo, 8IU treatment increased overt emotion salience (P=0.02, d=0.63). There was no statistically significant increase after 24IU treatment (P=0.12, d=0.4). The effects after 8IU oxytocin were observed despite no significant increase in peripheral blood plasma oxytocin concentrations. We found no significant effects for reading the mind in the eyes task performance or secondary outcome social-cognitive tasks (emotional dot probe and face-morphing). To our knowledge, this is the first trial to assess the dose-dependent effects of a single oxytocin administration in autism, with results indicating that a low dose of oxytocin can significantly modulate overt emotion salience despite minimal systemic exposure.


Effectiveness of Nanolime as a Stone Consolidant: A 4-Year Study of Six Common UK Limestones.
Protecting stone buildings from weathering and decay is a major challenge in the conservation of built heritage. Most of the stone consolidants currently available are well suited to silicate stones, but are less compatible with limestone. In this paper we present for the first time the results over a 4-year period of various consolidation treatments carried out using nanolime on 6 of the most representative and significant stones used in historic buildings in the United Kingdom. Tests investigated the influence of stone type, environmental conditions and pre-treatments on the effectiveness of the consolidation treatment. A comprehensive and rigorous testing programme was carried out to evaluate the short (12 weeks) and longer-term (4 years) effects. Stone samples were characterised before and after treatment using light and electron microscopy, sorptivity tests and a novel methodology employing drilling resistance to interrogate the near surface effects. Results show that for some of the stones, such as Clunch and Bath Stone, the positive effect of the treatment with nanolime is noticeable after 4 years since application. However, results for other stones such as Portland and magnesian limestone showed that the initial beneficial effect of the treatment is reduced after 4 years. Nanolime treatment of Ham Stone produced an unnoticeable effect on the continuous natural reduction of the drilling resistance of the specimen over time. The results presented are of immense value to conservators as they provide essential guidance on the most appropriate repair approach. Impact to the conservation industry will be to avoid the use of nanolime on stones where there is no perceivable benefit, reducing the risk of adverse effects, including potential damage to buildings. In additional costs will be saved which might otherwise have been spent on ineffective treatments.


Different dispersal abilities allow reef fish to coexist.
The coexistence of multiple species on a smaller number of limiting resources is an enduring ecological paradox. The mechanisms that maintain such biodiversity are of great interest to ecology and of central importance to conservation. We describe and prove a unique and robust mechanism for coexistence: Species that differ only in their dispersal abilities can coexist, if habitat patches are distributed at irregular distances. This mechanism is straightforward and ecologically intuitive, but can nevertheless create complex coexistence patterns that are robust to substantial environmental stochasticity. The Great Barrier Reef (GBR) is noted for its diversity of reef fish species and its complex arrangement of reef habitat. We demonstrate that this mechanism can allow fish species with different pelagic larval durations to stably coexist in the GBR. Further, coexisting species on the GBR often dominate different subregions, defined primarily by cross-shelf position. Interspecific differences in dispersal ability generate similar coexistence patterns when dispersal is influenced by larval behavior and variable oceanographic conditions. Many marine and terrestrial ecosystems are characterized by patchy habitat distributions and contain coexisting species that have different dispersal abilities. This coexistence mechanism is therefore likely to have ecological relevance beyond reef fish.


Neuronal and astroglial correlates underlying spatiotemporal intrinsic optical signal in the rat hippocampal slice.
Widely used for mapping afferent activated brain areas in vivo, the label-free intrinsic optical signal (IOS) is mainly ascribed to blood volume changes subsequent to glial glutamate uptake. By contrast, IOS imaged in vitro is generally attributed to neuronal and glial cell swelling, however the relative contribution of different cell types and molecular players remained largely unknown. We characterized IOS to Schaffer collateral stimulation in the rat hippocampal slice using a 464-element photodiode-array device that enables IOS monitoring at 0.6 ms time-resolution in combination with simultaneous field potential recordings. We used brief half-maximal stimuli by applying a medium intensity 50 Volt-stimulus train within 50 ms (20 Hz). IOS was primarily observed in the str. pyramidale and proximal region of the str. radiatum of the hippocampus. It was eliminated by tetrodotoxin blockade of voltage-gated Na(+) channels and was significantly enhanced by suppressing inhibitory signaling with gamma-aminobutyric acid(A) receptor antagonist picrotoxin. We found that IOS was predominantly initiated by postsynaptic Glu receptor activation and progressed by the activation of astroglial Glu transporters and Mg(2+)-independent astroglial N-methyl-D-aspartate receptors. Under control conditions, role for neuronal K(+)/Cl(-) cotransporter KCC2, but not for glial Na(+)/K(+)/Cl(-) cotransporter NKCC1 was observed. Slight enhancement and inhibition of IOS through non-specific Cl(-) and volume-regulated anion channels, respectively, were also depicted. High-frequency IOS imaging, evoked by brief afferent stimulation in brain slices provide a new paradigm for studying mechanisms underlying IOS genesis. Major players disclosed this way imply that spatiotemporal IOS reflects glutamatergic neuronal activation and astroglial response, as observed within the hippocampus. Our model may help to better interpret in vivo IOS and support diagnosis in the future.


ST analysis of the fetal electrocardiogram - Comments on recent experimental data.
In their paper, Andriessen at al present a validation of fetal ECG analysis and the clinical STAN device in midgestation fetal lambs exposed to 25 minutes of umbilical cord occlusion. The study presents results that contrast remarkably from previously published experimental data which raises a number of questions and comments. The most striking finding of Andriessen et al is the recording of an extremely high number of alarms from the STAN equipment during control conditions when no alarms at all are expected. These patterns have never been seen, neither in the clinical situation nor in our own fetal sheep studies. The reason for this becomes apparent when their way of recording the FECG is scrutinized. In their assessment of STAN, Andriessen at al use an assumed negative aVF lead with the assumption that it will reflect the FECG in the same way as the unipolar scalp lead used clinically. The signal used for disqualification of STAN is itself not qualified to properly represent the fetal scalp lead signal that STAN is designed for. To question a proven technology is fully accepted but those attempting would be asked to argue along fully validated data and related analysis including questioning of their own data.