Generation of humanized single-chain fragment variable immunotherapeutic against EGFR variant III using baculovirus expression system and in vitro validation.
Epidermal growth factor receptor variant III (EGFRvIII) is known to be specifically expressed in cancer cells and associated with tumor virulence. The receptor provides an opportunity for both specifically targeting the tumor cells as well as for potentially controlling and inhibiting tumor progression. In this study, humanized anti-EGFRvIII single-chain fragment variable (hscFv) was expressed in insect cell culture system to accommodate post-translational glycosylations crucial for the fragment stability and efficacy. Target specific binding of the developed fragment to EGFRvIII expressing cell lines and EGFRvIII positive glioblastoma patient samples was evaluated by immunocytochemistry and immunohistochemistry respectively. Downstream intracellular signaling mechanisms related to the action of the developed antibody fragment on growth/metabolism of the cell was evaluated in U87-EGFRvIII human glioblastoma cell lines. It was observed that the hscFv bound specifically to EGFRvIII in mutant expressing cells. Functionally, hscFv was found to confer anti-proliferative properties in EGFRvIII expressing cell lines by downregulating phosphorylation of EGFR/EGFRvIII, Lyn, PI3K and GLUT3 involved in proliferation and metabolism. This study demonstrated the significance of hscFv as a potential immunotherapeutic agent as well as a targeting agent for specific delivery of drugs to EGFRvIII expressing cancer cells.


[Communication on the Internet. Possibilities and risks of the Internet for psychiatry].
The article is preceded by an introduction to world-wide communication networks. Some important basic features are explained. The major impact is on temporary and future aspects of the Internet for psychiatry. A variety of advantages in World Wide Web telemedicine for patients, clients, psychotherapists, clinicians and scientists are extensively demonstrated. Useful and established possibilities are discussed. Finally, the risks of this kind of communication are shown and listed.


Single-cell RT-PCR and immunocytochemical detection of mechanosensitive transient receptor potential channels in acutely isolated rat odontoblasts.
Hydrostatic force applied to tooth pulp has long been suspected to be the direct cause of dental pain. However, the molecular and cellular identity of the transducer of the mechanical force in teeth is not clear. Growing number of literatures suggested that odontoblasts, secondary to its primary role as formation of tooth structure, might function as a cellular mechanical transducer in teeth. In order to determine whether odontoblasts could play a crucial role in transduction of hydrostatic force applied to dental pulp into electrical impulses, current study investigated the expression of stretch-activated transient receptor potential (TRP) channels in acutely isolated odontoblasts from adult rats by single cell reverse transcriptase polymerase chain reaction and immunocytochemical analysis. As the result, expression of TRPM7 (melastatin 7) was observed in majority (87%) of odontoblasts while mRNAs for TRPC1 (canonical 1), TRPC6 (canonical 6) and TRPV4 (vanilloid 4) were detected in small subpopulations of odontoblasts. TRPM3 (melastatin 3) was not detected in our experimental set-up. Immunocytochemical analysis further revealed TRPM7 expression at protein level. Expression of the mechanosensitive TRP channels provides additional evidence that supports the sensory roles of odontoblasts. Given that TRPM7 is a mechanosensitive ion channel with a kinase activity that plays a role in Mg(2+) homeostasis, it is possible that TRPM7 expressed in odontoblasts might play a central role in mineralization during dentin formation.


Inhibition of mutated, activated BRAF in metastatic melanoma.
The identification of somatic mutations in the gene encoding the serine-threonine protein kinase B-RAF (BRAF) in the majority of melanomas offers an opportunity to test oncogene-targeted therapy for this disease. We conducted a multicenter, phase 1, dose-escalation trial of PLX4032 (also known as RG7204), an orally available inhibitor of mutated BRAF, followed by an extension phase involving the maximum dose that could be administered without adverse effects (the recommended phase 2 dose). Patients received PLX4032 twice daily until they had disease progression. Pharmacokinetic analysis and tumor-response assessments were conducted in all patients. In selected patients, tumor biopsy was performed before and during treatment to validate BRAF inhibition. A total of 55 patients (49 of whom had melanoma) were enrolled in the dose-escalation phase, and 32 additional patients with metastatic melanoma who had BRAF with the V600E mutation were enrolled in the extension phase. The recommended phase 2 dose was 960 mg twice daily, with increases in the dose limited by grade 2 or 3 rash, fatigue, and arthralgia. In the dose-escalation cohort, among the 16 patients with melanoma whose tumors carried the V600E BRAF mutation and who were receiving 240 mg or more of PLX4032 twice daily, 10 had a partial response and 1 had a complete response. Among the 32 patients in the extension cohort, 24 had a partial response and 2 had a complete response. The estimated median progression-free survival among all patients was more than 7 months. Treatment of metastatic melanoma with PLX4032 in patients with tumors that carry the V600E BRAF mutation resulted in complete or partial tumor regression in the majority of patients. (Funded by Plexxikon and Roche Pharmaceuticals.)


Identification of electrostatic two-stream instabilities associated with a laser-driven collisionless shock in a multicomponent plasma.
Electrostatic two-stream instabilities play essential roles in an electrostatic collisionless shock formation. They are a key dissipation mechanism and result in ion heating and acceleration. Since the number and energy of the shock-accelerated ions depend on the instabilities, precise identification of the active instabilities is important. Two-dimensional particle-in-cell simulations in a multicomponent plasma reveal ion reflection and acceleration at the shock front, excitation of a longitudinally propagating electrostatic instability due to a nonoscillating component of the electrostatic field in the upstream region of the shock, and generation of up- and down-shifted velocity components within the expanding-ion components. A linear analysis of the instabilities for a C_{2}H_{3}Cl plasma using the one-dimensional electrostatic plasma dispersion function, which includes electron and ion temperature effects, shows that the most unstable mode is the electrostatic ion-beam two-stream instability (IBTI), which is weakly dependent on the existence of electrons. The IBTI is excited by velocity differences between the expanding protons and carbon-ion populations. There is an electrostatic electron-ion two-stream instability with a much smaller growth rate associated with a population of protons reflecting at the shock. The excitation of the fast-growing IBTI associated with laser-driven collisionless shock increases the brightness of a quasimonoenergetic ion beam.


Influence of electrical signals on pea leaf reflectance in the 400-800-nm range.
Local action of stressors induces generation and propagation of electrical signals (ESs), which influence numerous physiological processes (including photosynthesis, expression of genes, production of phytohormones, etc.) in undamaged parts of plants; i.e. they induce a systemic stress response. Development of methods of remote sensing of this response (in particular, optical methods) is an important practical task for agricultural and ecological monitoring. However, this problem is not sufficiently researched. Earlier, we reported that ESs influence the photochemical reflectance index, which can be calculated on the basis of reflected light at 531 and 570 nm, and these changes are connected with photosynthetic changes. The aim of the current work is investigation of the influence of ESs on reflectance at broad spectral bands (400-500 nm, 500-600 nm, 600-700 nm and 700-800 nm). We showed that burning-induced ESs caused transient increase of intensity of reflected light at the all investigated spectral bands of visible light: reflectance at 600-700 nm had the maximal magnitude of changes and reflectance at 700-800 nm had the minimal magnitude of changes. Dynamics of the reflectance changes were distinguished from dynamics of photosynthetic changes, induced by ESs; i.e. ESs-induced changes in reflectance seem to be weakly connected with the photosynthetic response. Thus, our results show that changes in reflectance at broad spectral bands can also be used for remote sensing of the ESs-induced systemic stress response in plants.


THE STIMULATION OF DNA SYNTHESIS IN CULTURES OF RABBIT LYMPH NODE AND SPLEEN CELL SUSPENSIONS BY HOMOLOGOUS CELLS.
An early proliferative response follows the mixing of homologous spleen or lymph node suspensions obtained from two unrelated rabbits. The rate of incorporation of radioactive thymidine has been used as a quantitative measure of this response. Thymus cells do not respond to homologous cell suspensions but may on occasion serve to stimulate the response in homologous spleen or lymph node cells. Homologous erythrocytes or autologous tissues do not stimulate a response. No response occurs if the two cell populations are separated by a Millipore membrane. Autoradiographic studies have established that 1 to 2 per cent of the intial cell population is involved in the response and they are large undifferentiated cells by the time they can first be identified as responders. There was no morphological evidence of any cellular interaction and the viability of mixed suspensions was not measurably different from that observed in separate suspension. Simultaneous additive responses could be obtained to homologous cells and to antigen when cell suspensions from immunized rabbits were used. The interaction of the cell populations from the two rabbits did not appear to suppress the response of each to antigen. The speed and magnitude of the response were in everyway comparable with the secondary response of cell suspensions from immunized rabbits exposed to the immunizing antigen. No evidence was obtained of any enhancement of the response to homologous cells by prior immunization with homologous tissues, but the possibility that it had occurred was not rigidly excluded.


HEPATITIS C TREATMENT OF RENAL TRANSPLANT AND CHRONIC KIDNEY DISEASE PATIENTS: EFFICACY AND SAFETY OF DIRECT-ACTING ANTIVIRAL REGIMENS CONTAINING SOFOSBUVIR.
Direct-acting antivirals have revolutionized hepatitis C treatment, also for patients with chronic kidney disease (CKD), but some controversy exists regarding the use of sofosbuvir (SOF) in patients with glomerular filtration rate (GFR) <30 mL/min. To evaluate the efficacy and safety of these regimens for hepatitis C treatment of patients with CKD and after renal transplantation, as well as the impact of SOF on renal function in non-dialysis patients. All patients with hepatitis C and CKD or renal transplant treated with direct-acting antivirals at a referral center in Brazil between January 2016 and August 2017 were included. Efficacy was evaluated based on viral load (HCV RNA) and a sustained virological response (SVR) consisting of undetectable RNA 12 and/or 24 weeks after the end of treatment (SVR12 and SVR24) was defined as cure. Safety was determined by adverse events and ribavirin, when combined, was administered in escalating doses to all patients with GFR <60 mL/min. The impact of SOF on renal function was determined by the measurement of baseline creatinine during and after the end of treatment and its increase was evaluated using the Acute Kidney Injury Network (AKIN) classification. A total of 241 patients (52.7% females) with a mean age of 60.72±10.47 years were included. The combination of SOF+daclatasvir was the predominant regimen in 75.6% of cases and anemia was present in 28% of patients who used ribavirin (P=0.04). The SVR12 and SVR24 rates were 99.3% and 97.1%, respectively. The treatment was well tolerated and there were no major clinically relevant adverse events, with the most prevalent being asthenia (57.7%), itching (41.1%), headache (40.7%), and irritability (40.2%). Among conservatively treated and renal transplant patients, oscillations of creatinine levels (AKIN I) were observed in 12.5% of cases during treatment and persisted in only 8.5% after the end of treatment. Of these, 2.0% had an initial GFR <30 mL/min and this percentage decreased to 1.1% after SOF use. Only 0.5% and 1.6% of the patients progressed to AKIN II and AKIN III elevation, respectively. The direct-acting antivirals were safe and efficacious in CKD patients treated with SOF-containing regimens, with the observation of high SVR rates, good tolerability and few severe adverse events. The combination with ribavirin increased the risk of anemia and the administration of escalating doses seems to be useful in patients with GFR <60 mL/min. In patients with GFR <30 mL/min, SOF had no significant renal impact, with serum creatinine returning to levels close to baseline after treatment.


Activities of superoxide dismutase and glutathione peroxidase in schizophrenic and manic-depressive patients.
Activities of superoxide dismutase (EC 184.108.40.206) and glutathione peroxidase (EC 22.214.171.124) in erythrocytes were evaluated in 50 schizophrenic and 20 manic-depressive patients, who were or were not being treated with different neuroleptic drugs, and results were compared with those for 58 normal individuals. Neuroleptic-treated and untreated schizophrenic patients showed similar activities of superoxide dismutase, about 60% higher than those found in normal individuals (p less than 0.001). In manic-depressive patients treated with either lithium (n = 8) or lithium plus neuroleptic drugs (n = 12), superoxide dismutase activities were increased by about 40% over those of normal subjects (p less than 0.001). Significantly abnormal activities of glutathione peroxidase were found only in the sub-group of schizophrenic women. These results are interpreted in terms of active oxygen species involvement in the psychiatric manifestations.


Focal adhesion assembly in myofibroblasts fosters a microenvironment that promotes tumor growth.
Cells within the tumor microenvironment influence tumor growth through multiple mechanisms. Pericytes such as hepatic stellate cells are an important cell within the tumor microenvironment; their transformation into highly motile myofibroblasts leads to angiogenesis, stromal cell recruitment, matrix deposition, and ensuing tumor growth. Thus, a better understanding of mechanisms that regulate motility of pericytes is required. Focal adhesions (FAs) form a physical link between the extracellular environment and the actin cytoskeleton, a requisite step for cell motility. FAs contain a collection of proteins including the Ena/VASP family member, vasodilator-stimulated phosphoprotein (VASP); however, a role for VASP in FA development has been elusive. Using a comprehensive siRNA knockdown approach and a variety of VASP mutants coupled with complementary cell imaging methodologies, we demonstrate a requirement of VASP for optimal development of FAs and cell spreading in LX2 liver myofibroblasts, which express high levels of endogenous VASP. Rac1, a binding partner of VASP, acts in tandem with VASP to regulate FAs. In vivo, perturbation of Ena/VASP function in tumor myofibroblast precursor cells significantly reduces pericyte recruitment to tumor vasculature, myofibroblastic transformation, tumor angiogenesis, and tumor growth, providing in vivo pathobiologic relevance to these findings. Taken together, our results identify Ena/VASP as a significant modifier of tumor growth through regulation of FA dynamics and ensuing pericyte/myofibroblast function within the tumor microenvironment.


Mediated electron transfer between Fe(II) adsorbed onto hydrous ferric oxide and a working electrode.
The redox properties of Fe(II) adsorbed onto mineral surfaces have been highly studied over recent years due to the wide range of environmental contaminants that react with this species via abiotic processes. In this work the reactivity of Fe(II) adsorbed onto hydrous ferric oxide (HFO) has been studied using ferrocene (bis-cyclopentadienyl iron(II); Fc) derivatives as electron shuttles in cyclic voltammetry (CV) experiments. The observed amplification of the ferrocene oxidation peak in CV is attributed to reaction between the electrochemically generated ferrocenium (Fc(+)) ion and adsorbed Fe(II) species in a catalytic process (EC' mechanism). pH dependence studies show that the reaction rate increases with Fe(II) adsorption and is maintained in the absence of aqueous Fe(2+), providing strong evidence that the electron transfer process involves the adsorbed species. The rate of reaction between Fc(+) and adsorbed Fe(II) increases with the redox potential of the ferrocene derivative, as expected, with bimolecular rate constants in the range 10(3)-10(5) M(-1) s(-1). The ferrocene-mediated electrochemical method described has considerable promise in the development of a technique for measuring electron-transfer rates in geochemical and environmental systems.


The PEP-pyruvate-oxaloacetate node: variation at the heart of metabolism.
At the junction between the glycolysis and the tricarboxylic acid cycle-as well as various other metabolic pathways-lies the phosphoenolpyruvate (PEP)-pyruvate-oxaloacetate node (PPO-node). These three metabolites form the core of a network involving at least eleven different types of enzymes, each with numerous subtypes. Obviously, no single organism maintains each of these eleven enzymes; instead, different organisms possess different subsets in their PPO-node, which results in a remarkable degree of variation, despite connecting such deeply conserved metabolic pathways as the glycolysis and the tricarboxylic acid cycle. The PPO-node enzymes play a crucial role in cellular energetics, with most of them involved in (de)phosphorylation of nucleotide phosphates, while those responsible for malate conversion are important redox enzymes. Variations in PPO-node therefore reflect the different energetic niches that organisms can occupy. In this review, we give an overview of the biochemistry of these eleven PPO-node enzymes. We attempt to highlight the variation that exists, both in PPO-node compositions, as well as in the roles that the enzymes can have within those different settings, through various recent discoveries in both bacteria and archaea that reveal deviations from canonical functions.


[Timing of bacterial colonization in severe burns: is strict isolation necessary?].
Infection is still one of the main causes of mortality in severe burn patients. Strict isolation has been used for the prevention of infection, but the efficacy of this measure is debatable. The aim of this study was to determine the timing of bacterial colonization in these patients and to ascertain whether strict isolation is indicated. Thirty consecutive children with severe burns were studied. Patients were only barrier-nursed during dressing changes. On admission and twice weekly over the entire hospital stay, burn, sputum, gastric aspirates, feces, and blood samples were obtained for culture. All isolates were tested for specific biotypes. Results were studied with linear regression and repeated measures ANOVA to determine the timing of colonization and cross-colonization between patients. On admission, normal cutaneous flora were isolated from burn cultures of all patients. The remaining cultures were negative. After one week, gastric aspirates were found to be colonized by gram-negative bacteria and fungi. This was followed by colonization of feces, burn, and sputum cultures. Biotype identification showed unidirectional colonization from the gastrointestinal tract to burns and upper airway. There were no cross infections between patients. Microbial colonization in severe burn patients was endogenous in nature and there were no cross infections. Thus, strict isolation is not necessary in burn centers, except during outbreaks of multi-resistant microorganisms.


Exercise Training and Neuromuscular Parameters in Patients With Type 1 Diabetes: Systematic Review and Meta-Analysis.
The present study aimed to systematically review the literature on the effects of physical training on neuromuscular parameters in patients with type 1 diabetes mellitus (T1DM). The PubMed, Scopus, EMBASE, and COCHRANE databases were accessed in September 2020. Clinical trials that evaluated the effects of physical training on neuromuscular parameters (maximum strength, resistance strength, muscle power, muscle quality, and muscle thickness) of patients with T1DM compared with a control group were considered eligible. The results were presented as the standardized mean difference with 95% confidence intervals. Effect size (ES) calculations were performed using the fixed effect method, with α = .05. Combined training increased the maximum strength levels in individuals with T1DM to a lesser extent than in healthy individuals (ES: 0.363; P = .038). Strength training increased the maximum strength (ES: 1.067; P < .001), as well as combined training (ES: 0.943; P < .001); both compared with aerobic training in patients with T1DM. Strength training increased the maximum strength in a similar magnitude to combined training in patients with T1DM (ES: -0.114; P = .624). Both combined training and strength training represent effective strategies for improving the maximum strength in individuals with T1DM.


Correlation of RECIST, Computed Tomography Morphological Response, and Pathological Regression in Hepatic Metastasis Secondary to Colorectal Cancer: The AVAMET Study.
Background: The prospective phase IV AVAMET study was undertaken to correlate response evaluation criteria in solid tumors (RECIST)-defined response rates with computed tomography-based morphological criteria (CTMC) and pathological response after liver resection of colorectal cancer metastases. Methods: Eligible patients were aged ≥18 years, with Eastern Cooperative Oncology Group (ECOG) performance status 0/1 and histologically-confirmed colon or rectal adenocarcinoma with measurable liver metastases. Preoperative treatment was bevacizumab (7.5 mg on day 1) + XELOX (oxaliplatin 130 mg/m2, capecitabine 1000 mg/m2 bid on days 1-14 q3w). After three cycles, response was evaluated by a multidisciplinary team. Patients who were progression-free and metastasectomy candidates received one cycle of XELOX before undergoing surgery 3-5 weeks later, followed by four cycles of bevacizumab + XELOX. Results: A total of 83 patients entered the study; 68 were eligible for RECIST, 67 for CTMC, and 51 for pathological response evaluation. Of these patients, 49% had a complete or partial RECIST response, 91% had an optimal or incomplete CTMC response, and 81% had a complete or major pathological response. CTMC response predicted 37 of 41 pathological responses versus 23 of 41 responses predicted using RECIST (p = 0.008). Kappa coefficients indicated a lack of correlation between the results of RECIST and morphological responses and between morphological and pathological response rates. Conclusion: CTMC may represent a better marker of pathological response to bevacizumab + XELOX than RECIST in patients with potentially-resectable CRC liver metastases.


Prevention of Incontinence-Associated Skin Damage in Nursing Homes.
Racial/ethnic disparities in preventing health problems have been reported in nursing homes. Incontinence is common among nursing home residents and can result in inflammatory-type skin damage, referred to as incontinence-associated skin damage (IASD). Little is known about the prevention of IASD and whether there are racial/ethnic disparities in its prevention. This study assessed the proportion of older nursing home residents receiving IASD prevention after developing incontinence after admission ( n = 10,713) and whether there were racial/ethnic disparities in IASD prevention. Predictors of preventing IASD were also examined. Four national data sets provided potential predictors at multiple levels. Disparities were analyzed using the Peters-Belson method; predictors of preventing IASD were assessed using hierarchical logistic regression. Prevention of IASD was received by 0.12 of residents and no racial/ethnic disparities were found. Predictors of preventing IASD were primarily resident-level factors including limitations in activities of daily living, poor nutrition, and more oxygenation problems.


Isolation and characterization of MOD5, a gene required for isopentenylation of cytoplasmic and mitochondrial tRNAs of Saccharomyces cerevisiae.
The mod5-1 mutation is a nuclear mutation in Saccharomyces cerevisiae that reduces the biosynthesis of N6-(delta 2-isopentenyl)adenosine in both cytoplasmic and mitochondrial tRNAs to less than 1.5% of wild-type levels. The tRNA modification enzyme, delta 2-isopentenyl pyrophosphate:tRNA isopentenyl transferase, cannot be detected in vitro with extracts from mod5-1 cells. A characterization of the MOD5 gene would help to determine how the same enzyme activity in different cellular compartments can be abolished by a single nuclear mutation. To that end we have cloned the MOD5 gene and shown that it restores delta 2-isopentenyl pyrophosphate:tRNA isopentenyl transferase activity and N6-(delta 2-isopentenyl)adenosine to tRNA in both the mitochondria and the nucleus/cytoplasm compartments of mod5-1 yeast cells. That MOD5 sequences are expressed in Escherichia coli and can complement an N6-(delta 2-isopentenyl)-2-methylthioadenosine-deficient E. coli mutant leads us to conclude that MOD5 is the structural gene for delta 2-isopentenyl pyrophosphate:tRNA isopentenyl transferase.


[Autism and attention deficit hyperactivity disorder: convergences and divergences. Genetics].
According to the DSM-5, attention deficit hyperactivity disorder (ADHD) and autism spectrum disorder (ASD) are different conditions that earlier versions of the DSM stated could not be diagnosed together in the same individual. Yet, over the last few decades the debate on the limits between the two disorders has continued, even though ADHD and ASD are undoubtedly clinically and cognitively different phenotypes, as demonstrated by the simple fact that they have been defined in clearly different ways. Thus, from a perspective anchored in a purely phenomenological view, there would be no grounds whatsoever on which to question the independence between the two disorders. Since, at the present time, the discussion on the convergence between ADHD and ASD cannot be considered to have been solved, this study aims to take the data available from genetics as the basis on which to review the nosological position of the two disorders. The main studies that have addressed this issue are reviewed. The data collected agree on a genetic overlap between ADHD and ASD, which is influenced by common molecular mechanisms that affect the two disorders at the same time. The conclusions that can be drawn from the data collected suggest a new conceptual model not only for ADHD and ASD, but also for complex mental disorders in general. This line of research will transform the way of understanding the treatment of mental disorders and will almost certainly open up new perspectives in this area.


Phase singularity of surface plasmon polaritons generated by optical vortices.
We demonstrate an experimental result that shows the phase singularity of surface plasmon waves generated by the direct transform of optical vortices at normal incidence focused on a structureless metal surface. The near-field two-dimensional intensity distribution near the focal plane is experimentally examined by using near-field scanning optical microscopy and shows a good agreement with the finite-difference time-domain simulation result. The experimental realization demonstrates a potential of the proposed excitation scheme to be reconfigured locally with advantages over structures milled into optically thick metallic films for plasmonics applications involving plasmonic vortices.


Defined pH-sensitive nanogels as gene delivery platform for siRNA mediated in vitro gene silencing.
In the present study, a pH sensitive nanogel platform for gene delivery was developed. The cationic nanogels based on dendritic polyglycerol (dPG) and low molecular weight polyethylenimine units were able to encapsulate siRNA during the manufacturing process. The thiol-Michael nanoprecipitation method, which operates under mild conditions and did not require any catalyst or surfactant, was used to develop tailor-made nanogels in the sub-100 nm range. The incorporation of pH sensitive benzacetal-bonds inside the nanogel network enables the controlled intracellular release of the cargo. The functionality to transport therapeutic biomolecules was tested by an in vitro GFP-siRNA transfection assay. Encapsulated siRNA could silence GFP expressing HeLa cells (up to 71% silencing in GFP). Furthermore, significantly reduced toxicity of the nanogel platform compared to the non-degradable PEI was observed. These properties realize a new carrier platform in the field of gene therapy.


Family size preference and factors affecting the fertility rate in Hyogo, Japan.
Japan has consistently shown a low fertility rate, which has been lower than the replacement level since 1974, and represents one of the least fertile countries in the world. This study was designed to determine the family size preference of and its effect on Japanese women. We conducted a questionnaire survey among women who visited the obstetrics and gynecology department of 18 hospitals and clinics in the Hyogo Prefecture, Japan, between October 2011 and February 2012. All the women were categorized according to age group and area of residence, and the survey results were statistically analyzed using a t test. A total of 1616 women were included in this study. There was no significant difference between the mean desired and actual marital ages (26.70 and 26.67 years, respectively). The mean desired number of children was 2.55, which was significantly more than the mean actual number of children (1.77) in all generations. The mean desired and actual numbers of children were more in the rural areas (2.73 and 2.09, respectively) than in the urban (2.54 and 1.70, respectively) and semi-urban areas (2.49 and 1.60, respectively). The mean number of family members was significantly greater in the rural areas (3.84) than in the urban (3.25) and semi-urban areas (3.05).The most important concern among women who had never delivered a baby was childbearing itself, followed by the expenses related to pregnancy and childbearing. The family size preference of the women in our study was higher than the actual numbers of children. The fertility intentions were low among the younger women but high among those living in rural areas with larger families.


Postpartum haemorrhage and risk of long-term hypertension and cardiovascular disease: an English population-based longitudinal study using linked primary and secondary care databases.
To investigate the long-term risk of developing hypertension and cardiovascular disease (CVD) among those women who suffered a postpartum haemorrhage (PPH) compared with those women who did not. Population-based longitudinal open cohort study. English primary care (The Health Improvement Network (THIN)) and secondary care (Hospital Episode Statistics (HES)) databases. Women exposed to PPH during the study period matched for age and date of delivery, and unexposed. We conducted an open cohort study using linked primary care THIN and HES Databases, from 1 January 1997 to 31 January 2018. A total of 42 327 women were included: 14 109 of them exposed to PPH during the study period and 28 218 matched for age and date of delivery, and unexposed to PPH. HRs for cardiovascular outcomes among women who had and did not have PPH were estimated after controlling for covariates using multivariate Cox regression models. Risk of hypertensive disease, ischaemic heart disease, heart failure, stroke or transient ischaemic attack. During a median follow-up of over 4 years, there was no significant difference in the risk of hypertensive disease after adjustment for covariates (adjusted HR (aHR): 1.03 (95% CI: 0.87 to 1.22); p=0.71). We also did not observe a statistically significant difference in the risk of composite CVD (ischaemic heart disease, heart failure, stroke or transient ischaemic attack) between the exposed and the unexposed cohort (aHR: 0.86 (95% CI: 0.52 to 1.43; p=0.57). Over a median follow-up of 4 years, we did not observe an association between PPH and hypertension or CVD.


Control of glycogen phosphorylase interconversion by phorbol esters, diacylglycerols, Ca2+ and hormones in isolated rat hepatocytes.
In isolated rat hepatocytes: phosphorylase activation by the ionophore A23187 was enhanced in the presence of tumour-promoting phorbol esters and 1,2- (but not 1,3-) diacylglycerols (dioleoyl- and oleoylacetyl-glycerol), with a similar dose-dependency; the activation of phosphorylase by phenylephrine (1 microM) (but not by vasopressin or glucagon) was inhibited both by tumour-promoting phorbol esters and diacylglycerols, but with a different dose-dependency: complete inhibition was achieved with concentrations of phorbol esters two orders of magnitude lower than those of diacylglycerol; binding of the alpha 1-adrenergic antagonist [3H]prazosin and its displacement by unlabelled prazosin was not significantly affected in the presence of the phorbol esters. The possible involvement of protein kinase C in the control of phosphorylase interconversion is discussed.


Effects of triple therapy with octreotide, galanin and serotonin on a human colon cancer cell line.
Human colon cancer cells were implanted subcutaneously into nude mice. After 12 days, the animals were divided into two groups. The first group received 40 microg/kg body weight of octreotide, galanin and serotonin via an intraperitoneally implanted pump. The second group received sterile saline only. Treatment lasted for 14 days. The volume and weight of the tumours in treated mice tended to decrease, though not with statistical significance. The proliferation index and the number of tumour blood vessels was significantly reduced in the mice given triple therapy. The apoptotic index, as detected by TUNEL method and monoclonal anti-poly (ADP-ribose) polymerase, was significantly higher in the treated mice. Though the results of this investigation are promising, it is uncertain as to what use the present findings may imply for the treatment of patients with colorectal cancer.


Identification and Discrimination of Salmonella enterica Serovar Gallinarum Biovars Pullorum and Gallinarum Based on a One-Step Multiplex PCR Assay.
Salmonella enterica serovar Gallinarum biovars Pullorum (S. Pullorum) and Gallinarum (S. Gallinarum) can result in pullorum disease and fowl typhoid in avian species, respectively, and cause considerable economic losses in poultry in many developing countries. Conventional Salmonella serotyping is a time-consuming, labor-intensive and expensive process, and the two biovars cannot be distinguished using the traditional serological method. In this study, we developed a rapid and reliable one-step multiplex polymerase chain reaction (PCR) assay to simultaneously identify and discriminate the biovars Pullorum and Gallinarum. The multiplex PCR method focused on three specific genes, stn, I137_08605 and ratA. Based on bioinformatics analysis, we found that gene I137_08605 was present only in S. Pullorum and S. Gallinarum, and a region of difference in ratA was deleted only in S. Pullorum after comparison with that of S. Gallinarum and other Salmonella serovars. Three pairs of primers specific for the three genes were designed for the multiplex PCR system and their selectivity and sensitivity were determined. The multiplex PCR results showed that S. Pullorum and S. Gallinarum could be identified and discriminated accurately from all tested strains including 124 strains of various Salmonella serovars and 42 strains of different non-Salmonella pathogens. In addition, this multiplex PCR assay could detect a minimum genomic DNA concentration of 67.4 pg/μL, and 100 colony forming units. The efficiency of the multiplex PCR was evaluated by detecting natural-occurring Salmonella isolates from a chicken farm. The results demonstrated that the established multiplex PCR was able to identify S. Gallinarum and S. Pullorum individually, with results being consistent with traditional serotyping and biochemical testing. These results demonstrated that a highly accurate and simple biovar-specific multiplex PCR assay could be performed for the rapid identification and discrimination of Salmonella biovars Gallinarum and Pullorum, which will be useful, particularly under massive screening situations.


Social justice and intercountry adoptions: the role of the U.S. social work community.
Using social justice as the conceptual foundation, the authors present the structural barriers to socially just intercountry adoptions (ICAs) that can exploit and oppress vulnerable children and families participating in ICAs. They argue that such practices threaten the integrity of social work practice in that arena and the survival of ICA as a placement option. Government structures, disparity of power between countries and families on both sides, perceptions regarding poverty, cultural incompetence, misconceptions about orphans and orphanages, lack of knowledge about the impact of institution-based care, and the profit motive are driving forces behind the growing shadow of unethical ICAs. The U.S. social work community has a large role and responsibility in addressing these concerns as the United States receives the most children adopted through ICAs of all receiving countries. In addition to the centrality of social justice as a core value of the profession, the responsibility to carry out ethical and socially just ICA has recently increased as a matter of law, under the implementation legislation to the Hague Convention on Intercountry Adoption. While acknowledging that these issues are complex, authors provide suggestions for corrective policy and practice measures.


Long-term surgical results of pubovaginal sling procedure using polypropylene mesh in the treatment of stress urinary incontinence.
Surgery for stress urinary incontinence (SUI) in women might cure incontinence symptom but develop new problems. This study assessed the long-term outcome of pubovaginal sling procedure using polypropylene mesh in patients with SUI. A total of 108 patients with various types of SUI received pubovaginal sling procedure using polypropylene mesh. A surgical technique that did not create bladder outlet obstruction after operation was used in all the patients. Video urodynamic study was performed at baseline, 2 weeks and 3 months after operation. The long-term surgical results and patients' satisfaction were assessed. The age of patients ranged from 33 to 94 years (mean 62.6 +/- 12.0 years). The follow-up period ranged from 24 to 72 months (median 46 months). Among the 108 patients, a successful result was noted in 92 (85.2%), including a continence rate of 65.7% and mild SUI in 19.4% of patients. Treatment failure was noted in 16 patients (14.8%), including 13 (12%) with urge incontinence and 3 (2.8%) with severe SUI that was cured by a second sling. There were 104 patients (96.3%) who could void volitionally with little residual urine, whereas 4 (3.7%) needed transvaginal urethrolysis. Urodynamic study revealed no significant changes in the mean values of parameters at 3 months after operation. However, 22 (20%) patients had an increase in voiding pressure by 50% at 3 months postoperatively. The reported satisfactory rate was 89.8%. Pubovaginal sling procedure using polypropylene mesh had a high success rate in all types of SUI. The continence rate was suboptimal, indicating that the absence of bladder outlet obstruction might result in inadequate urethral compression after pubovaginal sling procedure.


[Continuing medical education in Aesthetic Medicine: hands--on course on the usage of hyaluronic acids of different particle size on anatomic preparations].
The Division of Evidence Based Medicine (dEBM), Clinic for Dermatology, Venerology and Allergology, Charité--Universitätsmedizin Berlin, offers on a regular basis workshops focusing on different areas of aesthetic medicine. Once a year a joint course is provided in cooperation with the Institute of Anatomy, offering the participants the possibility to improve their injection techniques as well as their knowledge on the facial anatomy. This course is focused on treatment with hyaluronic acids of different particle size. Besides the classical indications, it considers new indications such as correcting the shape of the nose or lacrimal groove. Thirteen physicians participated in the course, which was evaluated as very helpful as it improved not only the injection technique but also the knowledge of anatomy.


Non-invasive Measurement of Brown Fat Metabolism Based on Optoacoustic Imaging of Hemoglobin Gradients.
Metabolism is a fundamental process of life. However, non-invasive measurement of local tissue metabolism is limited today by a deficiency in adequate tools for in vivo observations. We designed a multi-modular platform that explored the relation between local tissue oxygen consumption, determined by label-free optoacoustic measurements of hemoglobin, and concurrent indirect calorimetry obtained during metabolic activation of brown adipose tissue (BAT). By studying mice and humans, we show how video-rate handheld multi-spectral optoacoustic tomography (MSOT) in the 700-970 nm spectral range enables non-invasive imaging of BAT activation, consistent with positron emission tomography findings. Moreover, we observe BAT composition differences between healthy and diabetic tissues. The study consolidates hemoglobin as a principal label-free biomarker for longitudinal non-invasive imaging of BAT morphology and bioenergetics in situ. We also resolve water and fat components in volunteers, and contrast MSOT readouts with magnetic resonance imaging data.


Investigation of Key Factors for Accident Severity at Railroad Grade Crossings by Using a Logit Model.
Although several studies have used logit or probit models and their variants to fit data of accident severity on roadway segments, few have investigated accident severity at a railroad grade crossing (RGC). Compared to accident risk analysis in terms of accident frequency and severity of a highway system, investigation of the factors contributing to traffic accidents at an RGC may be more complicated because of additional highway-railway interactions. Because the proportional odds assumption was violated while fitting cumulative logit modeled by the proportional odds models with stepwise variable selection to ordinal accident severity data collected at 592 RGCs in Taiwan, as suggested by Strokes et al. (2000, p. 249) a generalized logit model with stepwise variable selection was used instead to identify explanatory variables (factors or covariates) that were significantly associated with the severity of collisions. Hence, the fitted model was used to predict the level of accident severity, given a set of values in the explanatory variables. Number of daily trains, highway separation, number of daily trucks, obstacle detection device, and approaching crossing markings significantly affected levels of accident severity at an RGC (p-value = 0.0009, 0.0008, 0.0112, 0.0017, and 0.0003, respectively). Finally, marginal effect analysis on the number of daily trains and law enforcement camera was conducted to evaluate the effect of the number of daily trains and presence of a law enforcement camera on the potential accident severity.


Maternal mortality in a teaching hospital in southern India. A 13-year study.
During the 13 years 1960-1972, in a teaching hospital that serves a predominantly rural and semiurban population in southern India, there were 74,384 deliveries and 1245 maternal deaths, a maternal mortality rate of 16.7 per 1000 births. Direct obstetric factors caused 854 (65.5%) of these deaths. The leading indirect or associated causes of maternal deaths were anemia, cerebrovascular accidents, and infectious hepatitis. During the past 13 years, monthly maternal mortality meetings have helped to reduce the incidence of avoidable factors in maternal deaths among patients from the city but not among those brought from the surrounding countryside. The important causes of maternal deaths in this developing country, and their prevention, are individually discussed.


Dissecting JNK signaling, one KKKinase at a time.
Specificity in signal transduction is essential to ensure distinct and appropriate cellular responses to extracellular cues. Determining the mechanisms that mediate specificity is key to understanding complex cell behaviors in development, when multiple pathways fire simultaneously and individual pathways are used recurrently. Jun kinase (JNK) signal transduction exemplifies a pathway that is used multiple times in animal development and homeostasis. Indeed, molecular genetic analysis of JNK signaling in Drosophila has shown that a core signaling module consisting of Hep (JNKK), Bsk (JNK), and Jun regulates various processes, including tissue morphogenesis, wound repair, stress response, innate immune response, and others. Six putative JNKK kinase (JNKKK) family members are present in the fly genome, which could activate the core module in response to distinct stimuli. The diversity of kinases at this level of the signaling hierarchy could substantially increase the number of possible signals that feed into activation of the core module. Recent studies have described the distinct phenotypic consequences of mutations in three of the genes, Slpr (dMLK), Tak1, and Mekk1. These data, together with Drosophila cell culture and genomic array analyses support the contention that the choice of JNKKK may contribute to signaling specificity in vivo. Whether this is achieved by individual JNKKKs or by means of a combinatorial mechanism will require a systematic characterization of compound mutants and a toolbox of transcriptional reporters specific for distinct JNK-dependent processes.


Childhood Obesity and Physical Activity-Friendly School Environments.
Childhood obesity may be related to school environment, but previous studies often focused on food environment only. This study aimed to examine the relationship between school physical activity environment and childhood obesity. This is a cross-sectional study with multilevel data collected on school physical activity environment using teacher questionnaires, students' growth, and obesity status from electronic health records, and neighborhood socioeconomic status from census data. This study included 208 280 students (6-18 years of age) from 438 schools (45% of Hong Kong). Prevalence of obesity was 5.0%. After controlling for socioeconomic status and intraschool correlation, robust Poisson regression revealed a reduced obesity risk associated with higher teachers' perceived physical activity benefits (risk ratio 0.96, 95% CI 0.94-0.99, P = .02), physical activity teaching experience (0.93, 0.91-0.96, P < .001), school campus size (0.93, 0.87-0.99, P = .02), physical activity ethos (0.91, 0.88-0.94, P < .001), number of physical activity programs (0.93, 0.90-0.96, P < .001), and physical activity facilities (0.87, 0.84-0.90, P < .001). Students in schools with at least 3 physical activity-friendly environmental factors (11.7%) had a much lower risk of obesity (0.68, 0.62-0.75, P < .001) than those without (23.7%). A physical activity-friendly school environment is associated with lower risk of obesity. School physical activity environment should be considered in future epidemiologic and intervention studies.


(5Z)-carbacyclin discriminates between prostacyclin-receptors coupled to adenylate cyclase in vascular smooth muscle and platelets.
(5E)- and (5Z)-carbacyclin are prostacyclin (PGI2) analogues endowed with antiaggregating and vasodilator properties, which stimulate adenylate cyclase activity in membranes from human platelets and cultured myocytes from rabbit mesenteric artery. In platelets they display the same efficacy as prostaglandin E1 (PGE1), and hence PGI2 both as activators of adenylate cyclase and as inhibitors of aggregation. In contrast, in vascular smooth muscle cells (5Z)-carbacyclin fails to produce the same degree of stimulation of the enzyme as PGI2, (5E)-carbacyclin and PGE1, nor does it induce the maximal relaxation of the mesenteric artery as do the other prostaglandins. (5Z)-carbacyclin is also able to antagonize the activation of adenylate cyclase and the relaxation elicited by PGE1 or PGI2 in the mesenteric artery, and therefore it displays partial agonist properties in these cells. We conclude that the receptors for PGI2 coupled to adenylate cyclase in platelets and vascular smooth muscle cells are different from each other, because (5Z)-carbacyclin can discriminate between them, being a partial agonist at myocyte but not at platelet level.


Folding and fibrillogenesis: clues from beta2-microglobulin.
Renal failure impairs the clearance of beta(2)-microglobulin from the serum, with the result that this protein accumulates in joints under the form of amyloid fibrils. While the molecular mechanism leading to deposition of amyloid in vivo is not totally understood, some organic compounds, such as trifluoroethanol (TFE), are commonly used to promote the elongation of amyloid fibrils in vitro. This article gives some insights into the structural properties and the conformational states of beta(2)-microglobulin in the presence of TFE, using both the wild-type protein and the mutant Trp60Gly. The structure of the native state of the protein is rather insensitive to the presence of the alcohol, but the stability of this state is lowered in comparison to some other conformational states. In particular, a native-like folding intermediate is observed in the presence of moderate concentrations of TFE. Instead, at higher concentrations of the alcohol, the population of a disordered native-unlike state is dominant and correlates with the ability to elongate fibrils.


Expression of cyclin D1 after treatment with doxorubicin in the HL-60 cell line.
Increased levels of cyclin D1 and amplification of CCND1 gene occur in many types of cancers. We have followed the expression of cyclin D1 after treatment with doxorubicin with reference to cell death and other possible therapeutic implications. The effect of the treatment on the cell cycle, survival, intracellular level (flow cytometry), and intracellular localization of cyclin D1 (fluorescence microscopy) and expression of CCND1 (real-time RT-PCR) was investigated in HL-60 cells. An increase in the fluorescence intensity of cyclin D1 occurred after treatment with 0.15 and 0.3 μM doxorubicin. This tendency was confirmed by real-time RT-PCR. Expression of CCND1 in relation to the reference gene PBGD was increased in cells exposed to 0.15 μM doxorubicin. Concomitantly, some alterations in the regulation of the G0/G1, S, and G2/M checkpoints occurred, accompanied by changes in the polyploid fraction of the population. This was particularly evident at 0.3 μM doxorubicin, at which concentration the rate of cell death was also clearly higher. In conclusion, depending on the concentration used, alterations in cell death and the number of S, G2/M, and polyploid cells may correspond with cyclin D1 levels. This, in turn, may reflect an important role of the protein as one of the possible survival/point-of-no-return regulators dependent on its concentration, which seems especially plausible in the context of more prominent cell death in the above-mentioned fractions of cells.


On the structural organization of isolated bovine lens fiber junctions.
Junctions between fiber cells of bovine lenses have been isolated in milligram quantities, without using detergents or proteases. The structure of the isolated junctions has been studied by thin-section, negative-stain, and freeze-fracture electron microscopy and by x-ray diffraction. The junctions are large and most often have an undulating surface topology as determined by thin sectioning and freeze-fracture. These undulations resemble the tongue-and-groove interdigitations between lens fiber cells previously seen by others (D. H. Dickson and G. W. Crock, 1972, Invest. Ophthalmol. 11:809-815). In sections, the isolated junctions display a pentalamellar structure approximately 13-14 nm in overall thickness, which is significantly thinner than liver gap junctions. Each junctional membrane contains in the plane of the lipid bilayers distinct units arranged in a square lattice with a center-to-center spacing of 6.6 nm. Freeze-fracture replicas of the junctions fractured transversely show that the repeating units extend across the entire thickness of each membrane. Each unit is probably constructed from four identical subunits, with each subunit containing a protein of an apparent molecular weight of 27,000. We conclude that the lens junctions are structurally and chemically, different from gap junctions and could represent a new kind of intercellular contact, not simply another crystalline state of the gap junction protein.


Antiganglioside antibodies in Guillain-Barré syndrome after a recent cytomegalovirus infection.
To study the association between anti-ganglioside antibody responses and Guillan-Barré syndrome (GBS) after a recent cytomegalovirus (CMV) infection. Enzyme linked immunosorbant assay (ELISA) was undertaken on serum samples from 14 patients with GBS with recent cytomegalovirus (CMV) infection (CMV+GBS) and 12 without (CMV-GBS), 17 patients with other neurological diseases (OND), 11 patients with a recent CMV infection but without neurological involvement, 11 patients with recent Epstein-Barr virus (EBV) infection but without neurological involvement, and 20 normal control (NC) subjects. IgM antibodies were found at 1:100 serum dilution to gangliosides GM2 (six of 14 patients), GM1 (four of 14), GD1a (three of 14) and GD1b (two of 14) in the serum samples of the CMV+GBS patients, but not in those of any of the CMV-GBS patients. IgM antibodies were also found to gangliosides GM1, GD1a, and GD1b in one of 11 OND patients, to ganglioside GM1 in one of 11 non- neurological CMV patients, and to ganglioside GD1b in one of 20 NC subjects. Some patients with EBV infection had IgM antibodies to gangliosides GM1 (five of 11), GM2 (three of 11), and GD1a (two of 11). However, the antibodies to ganglioside GM2 had a low titre, none being positive at 1:200 dilution, whereas five of the CMV+GBS serum samples remained positive at this dilution. Antibodies to ganglioside GM2 are often associated with GBS after CMV infection, but their relevance is not known. It is unlikely that CMV infection and anti-ganglioside GM2 antibodies are solely responsible and an additional factor is required to elicit GBS.


Single-Bead Quantification of Peptide Loading Distribution for One-Bead One-Compound Library Synthesis Using Confocal Raman Spectroscopy.
We report an analytical method to determine peptide loading of "one-bead one-compound" (OBOC) combinatorial peptide libraries at single-bead level. The quantification is based on a linear relationship between the amount of N-terminal amino groups on individual peptide beads and the intensity of Raman signal obtained from a specifically designed reporter labeled on amino groups. Confocal Raman spectroscopy was employed to characterize peptide loading of beads with defined peptide sequences and from OBOC combinatorial peptide libraries. Although amine loading of blank TentaGel beads was found to be uniform, peptide loading among beads of OBOC peptide libraries varied substantially, particularly for those libraries with long sequences. Construction of OBOC libraries can be monitored with this novel analytical technique so that synthetic conditions can be optimized for the preparation of high-quality OBOC peptide libraries. As the variability of peptide loading of individual library beads can significantly influence the screening results, quantitative information obtained by this method will allow us to gain insight into the complexity and challenge of OBOC library synthesis and screening.


Cultivation of Borrelia burgdorferi in dialysis membrane chambers in rat peritonea.
In order to be sustained within its enzootic cycle, B. burgdorferi must adapt to two strikingly different environments, the arthropod vector and the mammalian host. The ability to rapidly adapt to environmental changes is therefore presumed to be central to spirochete survival and pathogenic programs. Indeed, it has now been well established that tick feeding initiates extensive changes in both gene expression and protein composition, collectively referred to as "host adaptation," a process that is thought to continue throughout infection. The paucibacillary nature of borrelial infections, however, has hampered our ability to study this bacterium in vivo. To circumvent this limitation, an animal model was developed for obtaining sufficient numbers of organisms to directly examine differential gene expression and antigenic composition of B. burgdorferi within the context of the mammalian host. The DMC model allows for a direct comparison of host-adapted B. burgdorferi and their in vitro-cultivated counterparts.


Magnetic resonance imaging-guided laser interstitial thermal therapy as treatment for intractable insular epilepsy in children.
OBJECTIVE Seizure onset within the insula is increasingly recognized as a cause of intractable epilepsy. Surgery within the insula is difficult, with considerable risks, given the rich vascular supply and location near critical cortex. MRI-guided laser interstitial thermal therapy (LiTT) provides an attractive treatment option for insular epilepsy, allowing direct ablation of abnormal tissue while sparing nearby normal cortex. Herein, the authors describe their experience using this technique in a large cohort of children undergoing treatment of intractable localization-related epilepsy of insular onset. METHODS The combined epilepsy surgery database of Cook Children's Medical Center and Dell Children's Hospital was queried for all cases of insular onset epilepsy treated with LiTT. Patients without at least 6 months of follow-up data and cases preoperatively designated as palliative were excluded. Patient demographics, presurgical evaluation, surgical plan, and outcome were collected from patient charts and described. RESULTS Twenty patients (mean age 12.8 years, range 6.1-18.6 years) underwent a total of 24 LiTT procedures; 70% of these patients had normal findings on MRI. Patients underwent a mean follow-up of 20.4 months after their last surgery (range 7-39 months), with 10 (50%) in Engel Class I, 1 (5%) in Engel Class II, 5 (25%) in Engel Class III, and 4 (20%) in Engel Class IV at last follow-up. Patients were discharged within 24 hours of the procedure in 15 (63%) cases, in 48 hours in 6 (24%) cases, and in more than 48 hours in the remaining cases. Adverse functional effects were experienced following 7 (29%) of the procedures: mild hemiparesis after 6 procedures (all patients experienced complete resolution or had minimal residual dysfunction by 6 months), and expressive language dysfunction after 1 procedure (resolved by 3 months). CONCLUSIONS To their knowledge, the authors present the largest cohort of pediatric patients undergoing insular surgery for treatment of intractable epilepsy. The patient outcomes suggest that LiTT can successfully treat intractable seizures originating within the insula and offers an attractive alternative to open resection. This is the first description of LiTT applied to insular epilepsy and represents one of only a few series describing the use of LiTT in children. The results indicate that seizure reduction after LiTT compares favorably to that after conventional open surgical techniques.


Phosphorylation-dependent regulation of the guanylyl cyclase-linked natriuretic peptide receptor B: dephosphorylation is a mechanism of desensitization.
C-type natriuretic peptide (CNP) binds the guanylyl cyclase-linked natriuretic peptide receptor B (NPR-B) and stimulates marked elevations of the intracellular signaling molecule, cGMP. Here, the essential role of phosphorylation in the hormonal activation and deactivation of this receptor is described. Exposure of NIH3T3 fibroblasts overexpressing NPR-B (3T3-NPR-B) to CNP resulted in time-dependent decreases in both subsequent CNP-dependent cGMP elevations in whole cells and hormone-dependent guanylyl cyclase activity assayed in crude membranes. NPR-B isolated from resting 3T3-NPR-B cells was phosphorylated on serine and threonine residues, and exposure to CNP resulted in a time-dependent dephosphorylation and desensitization of the receptor. Immunoblot analysis and guanylyl cyclase activity assayed with the general activators Mn2+ and Triton X-100 indicated that these reductions were not due to receptor degradation. Tryptic phosphopeptide mapping analysis suggested that CNP treatment caused a complete dephosphorylation of approximately one-half of the NPR-B population. In vitro dephosphorylation of crude 3T3-NPR-B membranes with purified protein phosphatase 2A was highly correlated with losses in CNP- but not Mn2+- and Triton X-100-dependent guanylyl cyclase activity. Taken together, these data indicate that the catalytic activity of NPR-B is tightly coupled to its phosphorylation state and that dephosphorylation is a mechanism of desensitization.


Therapeutic bronchoscopy in the era of genotype directed lung cancer management.
Lung cancer is the leading cause of cancer related deaths. Non-small cell lung cancer (NSCLC) accounts for ~85% of lung cancers. Our understanding of driver mutations and genotype directed therapy has revolutionized the management of advanced NSCLC. Commonly described mutations include mutations in epidermal growth factor (EGFR) & BRAF and translocations in anaplastic lymphoma kinase (ALK) & rat osteosarcoma (ROS1). Drugs directed against these translocations have significantly improved progression free survival individually and have shown a survival benefit when studied in the Lung Cancer Mutation Consortium (median survival 3.5 vs. 2.4 years compared to standard therapy). In a related yet parallel universe, the number of bronchoscopic ablative modalities available for management of cancer related airway obstruction have increased exponentially over the past decade. A wealth of literature has given us a better understanding of the technical aspects, benefits and risks associated with these procedures. While they all show benefits in terms of relieving airway obstruction, symptom control, quality of life and lung function testing, their complication rates vary based on the modality. The overall complication rate was ~4% in the AQuIRE registry. Bronchoscopic therapeutic modalities include rigid bronchoscopy with mechanical debulking, laser, thermo-coagulation [electrocautery & argon plasma coagulation (APC)], cryotherapy, endobronchial brachytherapy (EBT), photodynamic therapy (PDT), intratumoral chemotherapy (ITC) and transbronchial needle injection (TBNI) of chemotherapy. Intuitively, one would assume that the science of driver mutations would crisscross with the science of bronchoscopic ablation as they overlap in the same patient population. Sadly, this is not the case and there is a paucity of literature looking at these fields together. This results in several unanswered questions about the interplay between these two therapies.


Karyotype evolution in South American subterranean rodents Ctenomys magellanicus (Rodentia: Octodontidae): chromosome rearrangements and (TTAGGG)n telomeric sequence localization in 2n=34 and 2n=36 chromosomal forms.
Ctenomys is the most numerous genus of South American subterranean rodents and one of the most karyotypically diverse clades of mammals known. Ctenomys magellanicus is the southernmost species of the group and the only one living in Isla Grande de Tierra del Fuego (Argentina). This species presents two chromosomal forms, i.e. 2n=34, and 2n=36 (FN=68). Recent studies suggest that genetic divergence between both karyotypic forms resulted from a chromosomal speciation process. In order to identify the chromosomal rearrangement involved in the process of karyotype evolution in this species, we used chromosome banding techniques and fluorescence in situ hybridization with a telomeric probe to metaphase chromosomes of the two chromosomal forms of Ctenomys magellanicus. Chromosome analysis of Giemsa-stained and G-banding preparations showed that Cm34 and Cm36 karyotypes differ in one rearrangement involving chromosomes A9 from Cm34 and B12 and B17 from Cm36. In addition FISH analysis showed that all of the chromosomes from both chromosomal forms exhibit a telomeric-only distribution pattern of the (TTAGGG)n sequence, indicating that none of the chromosomal forms of Ctenomys magellanicus has true telocentric chromosomes. Our results suggest that a chromosome fission event would have occurred during the process of karyotype evolution in this species.


Effects of benzyl isothiocyanate and 2-phenethyl isothiocyanate on benzo[a]pyrene and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone metabolism in F-344 rats.
A mixture of dietary benzyl isothiocyanate (BITC) and 2-phenethyl isothiocyanate (PEITC) inhibits lung tumorigenesis by a mixture of benzo[a]pyrene (B[a]P) and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in A/J mice. Previous studies indicated that inhibition of 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB) releasing DNA adducts of NNK by PEITC in the lung was responsible for inhibition of tumorigenicity. We have now extended these investigations to F-344 rats treated with 2 p.p.m. B[a]P in the diet and 2 p.p.m. NNK in the drinking water. The effects of BITC (1 micromol/g diet), PEITC (3 micromol/g diet), and a mixture of BITC plus PEITC (1 and 3 micromol/g diet) on DNA and hemoglobin (Hb) adducts of B[a]P and NNK, and on two urinary metabolites of NNK, were examined. DNA adducts were quantified after 8 and 16 weeks of treatment. Hb adducts were quantified in blood samples withdrawn every 2 weeks. 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) and its glucuronide NNAL-Gluc were measured in urine every 4 weeks. PEITC or BITC plus PEITC significantly reduced levels of HPB releasing DNA adducts of NNK in lung at 8 and 16 weeks, but there was no effect of BITC. There were no effects of any of the treatments on levels of HPB releasing DNA adducts of NNK in liver, or on DNA adducts of B[a]P in either lung or liver. PEITC or BITC plus PEITC significantly inhibited the formation of Hb adducts of NNK from 2-12 weeks of treatment while there were no effects on Hb adducts of B[a]P. There was a significant increase in levels of NNAL and NNAL-Gluc in the urine of the rats treated with PEITC or BITC plus PEITC. These results demonstrate that dietary PEITC, or a mixture of BITC plus PEITC, inhibit the formation of HPB releasing adducts of NNK in the rodent lung, leading to inhibition of tumorigenesis.


Complex tissue-specific patterns and distribution of multiple RAGE splice variants in different mammals.
The receptor for advanced glycosylation end products (RAGE) is a multiligand receptor involved in diverse cell signaling pathways. Previous studies show that this gene expresses several splice variants in human, mouse, and dog. Alternative splicing (AS) plays an important role in expanding transcriptomic and proteomic diversity, and it has been related to disease. AS is also one of the main evolutionary mechanisms in mammalian genomes. However, limited information is available regarding the AS of RAGE in a wide context of mammalian tissues. In this study, we examined in detail the different RAGE mRNAs generated by AS from six mammals, including two primates (human and monkey), two artiodactyla (cow and pig), and two rodentia (mouse and rat) in 6-18 different tissues including fetal, adult, and tumor. By nested reverse transcription-polymerase chain reaction (RT-PCR) we identified a high number of splice variants including noncoding transcripts and predicted coding ones with different potential protein modifications affecting mainly the transmembrane and ligand-binding domains that could influence their biological function. However, analysis of RNA-seq data enabled detecting only the most abundant splice variants. More than 80% of the detected RT-PCR variants (87 of 101 transcripts) are novel (different exon/intron structure to the previously described ones), and interestingly, 20-60% of the total transcripts (depending on the species) are noncoding ones that present tissue specificity. Our results suggest that RAGE undergoes extensive AS in mammals, with different expression patterns among adult, fetal, and tumor tissues. Moreover, most splice variants seem to be species specific, especially the noncoding variants, with only two (canonical human Tv1-RAGE, and human N-truncated or Tv10-RAGE) conserved among the six different species. This could indicate a special evolution pattern of this gene at mRNA level.


Real-time monitoring of in situ gas-phase H/D exchange reactions of cations by atmospheric pressure helium plasma ionization mass spectrometry (HePI-MS).
An enclosed atmospheric-pressure helium-plasma ionization (HePI-MS) source avoids, or minimizes, undesired back-exchange reactions usually encountered during deuterium incorporation experiments under ambient-pressure open-source conditions. A simple adaptation of an ESI source provides an economical way of conducting gas phase hydrogen/deuterium (H/D) exchange reactions (HDX) in real time without the need for complicated hardware modifications. For example, the spectrum of [(2)H8]toluene recorded under exposed ambient conditions showed the base peak at m/z 96 due to fast leaching of ring hydrogens because of interactions with H2O vapor present in the open source. Such D/H exchanges are rapidly reversed if the deuterium-depleted [(2)H8]toluene is exposed to D2O vapor. In addition to the enumeration of labile protons, our procedure enables the identification of protonation sites in molecules unambiguously, by the number of H/D exchanges observed in real time. For example, molecules such as tetrahydrofuran and pyridine protonate at the heteroatom and consequently undergo only one H/D exchange, whereas ethylbenzene, which protonates at a ring position of the aromatic ring, undergoes six H/D exchanges. In addition, carbocations generated in situ by in-source fragmentation of precursor protonated species, such as benzyl alcohol, do not undergo any rapid H/D exchanges. Because radical cations, second-generation cations (ions formed by losing a small molecule from a precursor ion), or those formed by hydride abstraction do not undergo rapid H/D exchanges, our technique provides a way to distinguish these ions from protonated molecules.


Gender differences in variance and means on the Naglieri Non-verbal Ability Test: data from the Philippines.
Research on gender differences in intelligence has focused mostly on samples from Western countries and empirical evidence on gender differences from Southeast Asia is relatively sparse. This article presents results on gender differences in variance and means on a non-verbal intelligence test using a national sample of public school students from the Philippines. More than 2,700 sixth graders from public schools across the country were tested with the Naglieri Non-verbal Ability Test (NNAT). Variance ratios (VRs) and log-transformed VRs were computed. Proportion ratios for each of the ability levels were also calculated and a chi-square goodness-of-fit test was performed. An analysis of variance was performed to determine the overall gender difference in mean scores as well as within each of three age subgroups. Our data show non-existent or trivial gender difference in mean scores. However, the tails of the distributions show differences between the males and females, with greater variability among males in the upper half of the distribution and greater variability among females in the lower half of the distribution. Descriptions of the results and their implications are discussed. Results on mean score differences support the hypothesis that there are no significant gender differences in cognitive ability. The unusual results regarding differences in variance and the male-female proportion in the tails require more complex investigations.


Integrin-linked kinase and ELMO2 modulate recycling endosomes in keratinocytes.
The formation of tight cell-cell junctions is essential in the epidermis for its barrier properties. In this tissue, keratinocytes follow a differentiation program tightly associated with their movement from the innermost basal to the outer suprabasal layers, and with changes in their cell-cell adhesion profile. Intercellular adhesion in keratinocytes is mediated through cell-cell contacts, including E-cadherin-based adherens junctions. Although the mechanisms that mediate E-cadherin delivery to the plasma membrane have been widely studied in simple epithelia, this process is less well understood in the stratified epidermis. In this study, we have investigated the role of Engulfment and Cell Motility 2 (ELMO2) and integrin-linked kinase (ILK) in the positioning of E-cadherin-containing recycling endosomes during establishment of cell-cell contacts in differentiating keratinocytes. We now show that induction of keratinocyte differentiation by Ca2+ is accompanied by localization of ELMO2 and ILK to Rab4- and Rab11a-containing recycling endosomes. The positioning of long-loop Rab11a-positive endosomes at areas adjacent to cell-cell contacts is disrupted in ELMO2- or ILK-deficient keratinocytes, and is associated with impaired localization of E-cadherin to cell borders. Our studies show a previously unrecognized role for ELMO2 and ILK in modulation of endosomal positioning, which may play key roles in epidermal sheet maintenance and permeability barrier function.


Structure of (+)-[S-(E)]-N-(alpha-methylbenzylidene)-p-toluenesulfinamide.
C15H15NOS, Mr = 257.35, monoclinic, P2(1), a = 7.710 (2), b = 5.903 (5), c = 14.700 (3) A, beta = 91.48 (2) degrees, V = 668.8 (6) A3, Z = 2, Dx = 1.278 g cm-3, lambda(Mo K alpha) = 0.71069 A, mu = 2.18 cm-1, F(000) = 272, T = 296 K, R = 0.043, 944 unique observed reflections. This is the first structure determination of an N-alkylidenesulfinamide. The C = N bond is distorted and lengthened presumably because of the steric interaction and electron-withdrawing effect of the sulfinyl group. The nonplanar conformation is also a result of repulsive steric interaction. The configuration at C = N is trans. The title compound was obtained from the displacement reaction of (-)-1-methyl (S)-p-toluenesulfinate with methyllithium.


Preferred orientations of His64 in human carbonic anhydrase II.
Histidine at position 64 (His64) in human carbonic anhydrase II (HCA II) is believed to be the proton acceptor in the hydration direction and the proton donor in the dehydration direction for the rate-limiting proton transfer (PT) event. Although the biochemical effect of histidine at position 64 has been thoroughly investigated, the role of its orientation in the PT event is a topic of considerable debate. X-ray data of HCA II suggests that His64 can adopt either an "in" or "out" orientation. The "in" orientation is believed to be favored for the hydration direction PT event because the Ndelta of His64 is closer to the catalytic zinc. This orientation allows for smaller water bridges, which are postulated to be more conducive to PT. In the present work, classical molecular dynamics simulations have been conducted to elucidate the role that the His64 orientation may play in its ability to act as a proton donor/acceptor in HCA II. The free energy profile for the orientation of His64 suggests that the histidine will adopt an "in" orientation in the hydration direction, which brings Ndelta in close proximity to the catalytic zinc. When the histidine becomes protonated, it then rotates to an "out" orientation, creating a more favorable solvation environment for the protonated His64. In this "out" orientation, the imidazole ring releases the delta nitrogen's excess proton into the bulk environment. After the second PT event and when the zinc-bound water is regenerated, the His64 is again favored to reorient to the "in" orientation, completing the catalytic cycle.


Facial interpretation and component consistency.
The issue of consistency, without regard to accuracy, of the facial component of focus (upper/eyebrows, middle/eyes, lower/nose and mouth) when interpreting facial expressions of emotion was examined. Eighty girls (average age, 7 years) chose, from an array of schematic faces, which face best represented various emotional scenarios (anger, disgust, fear, happiness, sadness, and surprise). The findings indicate that these children were remarkably consistent in facial area of focus. Across six trials for each emotion, children focused on the mouth (lower component) for happiness, sadness, surprise, and disgust, and on the eyebrows (the upper component) for anger and fear.


Development of a device for the delivery of agents to bone during distraction osteogenesis.
Various agents have been theoretically and experimentally implicated as mediators of distraction osteogenesis (DO). The purpose of this study was to develop a vehicle for the potential delivery of these factors to the region of the distraction site in an attempt to manipulate this biologic process. Three adult mongrel dogs (12 months old) had oblique osteotomies performed bilaterally through the gonial regions. In group I, the external distracter was affixed to the right hemimandible of two dogs (n = 2 hemimandibles) with cannulated pins (external diameter = 1.5 mm; lumen diameter = 1.0 mm; length = 60 mm), whereas the distracter on the left was affixed with standard, noncannulated pins of the same dimensions. In group II, cannulated pins were used to affix the external distracter to both hemimandibles (n = 2 hemimandibles) of a dog. The devices were activated after a 5-day latency period and were lengthened at a rate of 1 mm/day for 20 days. During the distraction period, 0.1 ml/d of sterile india ink was injected into the cannulated pins, after which the sterile stylet was replaced. The activation protocol was followed by 28 days of fixation (consolidation period). The hemimandibles from group I underwent removal of soft tissues, acetone fixation, and gross examination/photography, whereas the hemimandibles from group II were prepared for histologic evaluation (whole mount, hematoxylin and eosin staining). All dogs survived to the end of the study and demonstrated successful DO without evidence of complications. Hemimandibles in group I displayed evidence of india ink on both the lingual and buccal cortex around the cannulated pin site, in the regenerate and on the neocortices of the distracted segment. Hemimandibles of group II showed histologic evidence of the india ink being deposited densely around the cannulated pin site and extending in a radial fashion around the pin site into the regenerate. This study demonstrates for the first time a vehicle device for the delivery of an inert dye to the regenerate site during distraction osteogenesis. This vehicle offers the potential of delivery of various factors implicated in distraction osteogenesis (i.e., mitogens) in an attempt to alter this process and also substances (i.e., chemotherapy, antibiotics, etc.) for use in the treatment of various osteopathies.


Beach landscape Dataset of Fernando de Noronha Island (Brazil).
Beach landscape Dataset of Fernando de Noronha Island (Brazil), using a checklist with 26 physical and human parameters. Fernando de Noronha beaches were divided into sectors according to the landscape diversity. In total, 19 sectors were evaluated based on observations done during walks in the area, observations from viewpoints, with remote data. The evaluations were performed during fieldwork from 2014 (summer) and 2016 (spring). The landscape quality evaluation of Fernando de Noronha was performed using the Coastal Scenery Evaluation System. This method converts qualitative-quantitative data in quantitative data by estimating weights for 26 parameters (18 physical parameters - P - and 8 human-related parameters - H). The main parameters that define the landscape quality are classified from 1 (absence/bad quality) to 5 (presence/excellent quality). A mathematical model based on fuzzy logic was utilized to integrate the parameters weights in a special system for the scenarios classifications resulting in a value named D. The D-value is the indicator of the attractiveness of the evaluated place. The beaches are divided into classes ranging from 1 (extremely attractive natural site) to 5 (unattractive urban areas).


Solution structure of the RNA-binding cold-shock domain of the Chlamydomonas reinhardtii NAB1 protein and insights into RNA recognition.
Light-harvesting complex (LHC) proteins are among the most abundant proteins on Earth and play critical roles in photosynthesis, both in light capture and in photoprotective mechanisms. The Chlamydomonas reinhardtii nucleic acid-binding protein 1 (NAB1) is a negative regulator of LHC protein translation. Its N-terminal cold-shock domain (CSD) binds to a 13-nt element [CSD consensus sequence (CSDCS)] found in the mRNA of specific LHC proteins associated with Photosystem II (PSII), an interaction which regulates LHC expression and, consequently, PSII-associated antenna size, structure and function. In the present study, we elucidated the solution structure of the NAB1 CSD as determined by heteronuclear NMR. The CSD adopts a characteristic five-stranded anti parallel β-barrel fold. Upon addition of CSDCS RNA, a large number of NMR chemical shift perturbations were observed, corresponding primarily to surface-exposed residues within the highly conserved β2- and β3-strands in the canonical RNA-binding region, but also to residues on β-strand 5 extending the positive surface patch and the overall RNA-binding site. Additional chemical shift perturbations that accompanied RNA binding involved buried residues, suggesting that transcript recognition is accompanied by conformational change. Our results indicate that NAB1 associates with RNA transcripts through a mechanism involving its CSD that is conserved with mechanisms of sequence-specific nucleic acid recognition employed by ancestrally related bacterial cold-shock proteins (CSPs).


Inhibition of bone resorption by inorganic phosphate is mediated by both reduced osteoclast formation and decreased activity of mature osteoclasts.
High concentrations of inorganic phosphate (Pi) are known to inhibit bone resorption, although the mechanism(s) underlying this effect is unclear. To investigate whether Pi can inhibit the formation of osteoclasts we studied the effects of changes in Pi concentration between 1 and 4 mM on osteoclast-like cell formation in 1 week cultures of mouse bone marrow. Osteoclast-like cells were identified by multinuclearity, positive staining for tartrate-resistant acid phosphatase (TRAP), and contraction in response to calcitonin. Increasing concentrations of Pi inhibited formation of these cells in a dose-dependent manner. To study effects of Pi on the bone-resorbing activity of mature osteoclasts we isolated osteoclasts from calcium-deficient egg-laying hens or rat pups and incubated them on sperm whale dentine slices. High Pi concentrations markedly reduced both the number of resorption pits formed per dentine slice and the mean area of each pit in both avian and mammalian systems. These data indicate that high concentrations of Pi act on bone directly, both to inhibit generation of new osteoclasts from their precursor cells and to inhibit bone resorption by mature osteoclasts. These effects of extracellular Pi concentration may play an important modulatory role on bone turnover in vivo and have potential importance in several disease states in which Pi metabolism is perturbed.


Orthotopic bladder substitution (Neobladder): part II: postoperative complications, management, and long-term follow-up.
Bladder replacement following radical cystectomy is widely practiced and in some centers has become the standard method of urinary diversion when possible, rather than the traditional ileal conduit. To minimize the impact of cystectomy and bladder replacement on quality of life and to obtain optimal functional results, postoperative care must be comprehensive and multidisciplinary. Critical team members include the surgeon, urologic nurses, WOC nurses, and allied health care providers such as physiotherapists. This article highlights postoperative considerations for patients undergoing radical cystectomy with orthotopic neobladder construction. The common issues and complications that arise are discussed with a focus on strategies to optimize outcomes.


When eukaryotes and prokaryotes look alike: the case of regulatory RNAs.
The discovery that all living entities express many RNAs beyond mRNAs, tRNAs and rRNAs has been a surprise in the past two decades. In fact, regulatory RNAs (regRNAs) are plentiful, and we report stunning parallels between their mechanisms and functions in prokaryotes and eukaryotes. For instance, prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats) defense systems are functional analogs to eukaryotic RNA interference processes that preserve the cell against foreign nucleic acid elements. Regulatory RNAs shape the genome in many ways: by controlling mobile element transposition in both domains, via regulation of plasmid counts in prokaryotes, or by directing epigenetic modifications of DNA and associated proteins in eukaryotes. RegRNAs control gene expression extensively at transcriptional and post-transcriptional levels, with crucial roles in fine-tuning cell environmental responses, including intercellular interactions. Although the lengths, structures and outcomes of the regRNAs in all life kingdoms are disparate, they act through similar patterns: by guiding effectors to target molecules or by sequestering macromolecules to hamper their functions. In addition, their biogenesis processes have a lot in common. This unifying vision of regRNAs in all living cells from bacteria to humans points to the possibility of fruitful exchanges between fundamental and applied research in both domains.


Cancer-Associated Fibroblasts Enhance Survival and Progression of the Aggressive Pancreatic Tumor Via FGF-2 and CXCL8.
Pancreatic ductal adenocarcinoma remains one of the most challenging human cancers. Desmoplasia is predominant in this disease exhibiting a strong stromal reaction with an abundance of the cancer-associated fibroblasts (CAFs). We aimed in this study to investigate the reciprocal interaction between the tumor cells and the CAFs and its effect on tumor cells survival. We hypothesized that the survival of pancreatic cancer cell with aggressive phenotype is modulated by the Interactions between malignant pancreatic tumor cells and surrounding CAFs. To examine this, we utilized co-culture methods where tumor cells with different malignant potentials, HPAF (low) HPAF-CD11 (moderate/high) co-cultured with CAFs. CAFs-conditioned media increased the growth of HPAF-CD11 but not HPAF cells and increased CXCL8 levels highly in HPAF-CD11 and slightly in HPAF. The growth stimulatory effect and elevated CXCL8 level caused by CAFs-conditioned media were diminished by neutralizing the fibroblast growth factor-2 (FGF-2). In addition, conditioned media of HPAF-CD11 increased CAFs cell number whereas that of HPAF did not, and these effects were suppressed by neutralizing CXCL8. Furthermore, data from gene expression microarray study exhibited different expression profiles between HPAF and HPAF-CD11 when co-culture with CAFs. A significant increase in CXCL8 and FGF-2 expression was observed with HPAF-CD11/CAFs co-culture and to a lower extent with HPAF/CAFs co-culture. Together, these data demonstrate a paracrine bi-directional interaction between pancreatic tumor cells and the CAFs through CXCL8 and FGF-2 that helps the tumor growth. Future in-depth study of these pathways will assist in obtaining diagnostic and therapeutic tools for pancreatic ductal adenocarcinoma.


Functional MRI study of working memory in MDMA users.
Methylene-dioxymethamphetamine (MDMA) is known to cause degeneration of serotonin nerve terminals after acute doses in animals. Similarly, behavioral studies in human MDMA users regularly find abnormalities in memory, mood, and impulse control. However, studies of brain function using brain imaging in MDMA users have been less consistent. The purpose of this study was to determine, using functional magnetic resonance imaging (fMRI), whether individuals with a self-reported history of MDMA use would differ from non-MDMA using controls on activation while performing a working memory task. Fifteen MDMA using subjects and 19 non-MDMA using controls underwent fMRI scanning while performing the immediate and delayed memory task (IMT/DMT). The study was based on a block design in which the delayed memory task (DMT) alternated with the immediate memory task (IMT), which served as a control condition. FMRI scans were acquired on a 1.5 T scanner, using a gradient echo echoplanar pulse sequence. Random effects SPM99 analysis showed significantly greater activation (whole volume corrected cluster P<0.05) during the DMT relative to the IMT in the MDMA subjects compared with the control subjects in the medial superior frontal gyrus, in the thalamus extending into putamen, and in the hippocampus. Although these effects could be due to other drugs used by MDMA users, these results are consistent with behavioral problems that are associated with MDMA use, and with animal studies on the effects of MDMA on brain function.


Lipid domains in model membranes: a brief historical perspective.
All biological membranes consist of a complex composite of macromolecules and macromolecular assemblies, of which the fluid lipid-bilayer component is a core element with regard to cell encapsulation and barrier properties. The fluid lipid bilayer also supports the functional machinery of receptors, channels and pumps that are associated with the membrane. This bilayer is stabilized by weak physical and colloidal forces, and its nature is that of a self-assembled system of amphiphiles in water. Being only approximately 5 nm in thickness and still encapsulating a cell that is three orders of magnitude larger in diameter, the lipid bilayer as a material has very unusual physical properties, both in terms of structure and dynamics. Although the lipid bilayer is a fluid, it has a distinct and structured trans-bilayer profile, and in the plane of the bilayer the various molecular components, viz different lipid species and membrane proteins, have the capacity to organize laterally in terms of differentiated domains on different length and time scales. These elements of small-scale structure and order are crucial for the functioning of the membrane. It has turned out to be difficult to quantitatively study the small-scale structure of biological membranes. A major part of the insight into membrane micro- and nano-domains and the concepts used to describe them have hence come from studies of simple lipid bilayers as models of membranes, by use of a wide range of theoretical, experimental and simulational approaches. Many questions remain to be answered as to which extent the result from model studies can carry over to real biological membranes.


Dephosphorylation of cell cycle-regulated proteins correlates with anoxia-induced suspended animation in Caenorhabditis elegans.
Some metazoans have evolved the capacity to survive severe oxygen deprivation. The nematode, Caenorhabditis elegans, exposed to anoxia (0 kPa, 0% O(2)) enters into a recoverable state of suspended animation during all stages of the life cycle. That is, all microscopically observable movement ceases including cell division, developmental progression, feeding, and motility. To understand suspended animation, we compared oxygen-deprived embryos to nontreated embryos in both wild-type and hif-1 mutants. We found that hif-1 mutants survive anoxia, suggesting that the mechanisms for anoxia survival are different from those required for hypoxia. Examination of wild-type embryos exposed to anoxia show that blastomeres arrest in interphase, prophase, metaphase, and telophase but not anaphase. Analysis of the energetic state of anoxic embryos indicated a reversible depression in the ATP to ADP ratio. Given that a decrease in ATP concentrations likely affects a variety of cellular processes, including signal transduction, we compared the phosphorylation state of several proteins in anoxic embryos and normoxic embryos. We found that the phosphorylation state of histone H3 and cell cycle-regulated proteins recognized by the MPM-2 antibody were not detectable in anoxic embryos. Thus, dephosphorylation of specific proteins correlate with the establishment and/or maintenance of a state of anoxia-induced suspended animation.


[Relations between physical symptoms and humor].
In a study with 80 male non-psychology students we investigated the relationship between body-symptoms and the different kinds of humour-appreciation. It was hypothesized that the body-symptoms are correlated to humour. The hypotheses were related to both components in appreciation of humor, funniness (representing the positive feelings induced by humour) and aversiveness (representing the negative feelings). We can demonstrate, that body-symptoms are not related to funniness but to aversiveness of humour.


Laparoscopic Removal of an Abdominal Pregnancy in the Pelvic Sidewall.
To describe the presentation, diagnosis, and management of a patient with abdominal pregnancy and to illustrate the laparoscopic technique used to manage this patient. A descriptive study approved by our local Institutional Review Board. Consent was given from the patient. A university hospital in Ashdod, Israel. On May 15th 2018, a gravida 3, para 2, 37-year-old asymptomatic patient was referred to our hospital's gynecologic emergency department due to a suspected ectopic pregnancy. The patient had no relevant medical or surgical history. Her obstetric history consisted of 2 spontaneous vaginal deliveries with no other significant gynecologic history. Her menses were regular every month. Her last menstrual period was 6 weeks before presentation. Her β-human chorionic gonadotropin (hCG) level measured on the day before presentation was 24,856 IU/L. Physical examination was unremarkable except for a small amount of brownish vaginal discharge. A transvaginal ultrasound (TVUS) exam on presentation did not demonstrate an intrauterine gestational sac, but revealed a gestational sac and a fetus next to the right adnexa, with a crown-rump length of 1.3 cm, consistent with 7+3 gestational weeks. There was a minimal amount of fluid in the pouch of Douglas. Owing to an extrauterine pregnancy with high β-hCG value, laparoscopic operative management was chosen. On entrance to the abdominal cavity, a normal uterus and 2 ovaries and fallopian tubes were observed. A small to moderate amount of blood was present in the pouch of Douglas. a 3- to 4-cm distension was noted over the right uterosacral ligament. Following delicate probing of the area, moderate to severe bleeding commenced, which was initially controlled with local pressure and oxidized regenerated cellulose. An intraoperative TVUS identified an abdominal pregnancy in the right pelvic sidewall. The gestational sac was completely dissected and removed following ureterolysis and separation of the right ureter from the specimen. Local injection of vasopressin was also used. The β-hCG level before surgery of 19,008 IU/L decreased to 6339 IU/L on postoperative day 1. The patient was discharged in good condition on postoperative day 2. A final histopathological report confirmed a gestational sac. This patient referred for a tubal ectopic pregnancy was eventually diagnosed with an abdominal pregnancy and was treated operatively with complete excision of the abdominal pregnancy, which was located at the right pelvic sidewall. Abdominal pregnancy is a rare type of ectopic pregnancy with a reported incidence of 1:10,000 to 1:30,000 pregnancies, and accounts for approximately 1% of ectopic pregnancies [1]. It carries a high risk for maternal morbidity and mortality. Many different locations at different gestational ages have been reported in the literature, including the pouch of Douglas, pelvic sidewall, bowel, broad ligament, omentum, and spleen [2-4]. These varied locations and the rarity of this type of pregnancy make diagnosis and treatment challenging. The location of the growing fetal tissue may endanger the patient's life if it impinges on vital anatomic structures. In the present case, the gestational sac was very close to the right ureter, and we opted to surgically excise the gestational sac in its entirety.


Advances in neuroimaging of traumatic brain injury and posttraumatic stress disorder.
Improved diagnosis and treatment of traumatic brain injury (TBI) and posttraumatic stress disorder (PTSD) are needed for our military and veterans, their families, and society at large. Advances in brain imaging offer important biomarkers of structural, functional, and metabolic information concerning the brain. This article reviews the application of various imaging techniques to the clinical problems of TBI and PTSD. For TBI, we focus on findings and advances in neuroimaging that hold promise for better detection, characterization, and monitoring of objective brain changes in symptomatic patients with combat-related, closed-head brain injuries not readily apparent by standard computed tomography or conventional magnetic resonance imaging techniques.