Peripheral obestatin has no effect on feeding behavior and brain Fos expression in rodents.
Obestatin is produced in the stomach from proghrelin by post-translational cleavage. The initial report claimed anorexigenic effects of obestatin in mice. Contrasting studies indicated no effect of obestatin on food intake (FI). We investigated influences of metabolic state (fed/fasted), environmental factors (dark/light phase) and brain Fos response to intraperitoneal (ip) obestatin in rats, and used the protocol from the original study assessing obestatin effects in mice. FI was determined in male rats injected ip before onset of dark or light phase, with obestatin (1 or 5 micromol/kg), CCK8S (3.5 nmol/kg) or 0.15 M NaCl, after fasting (16 h, n=8/group) or ad libitum (n=10-14/group) food intake. Fos expression in hypothalamic and brainstem nuclei was examined in freely fed rats 90 min after obestatin (5 micromol/kg), CCK8S (1.75 nmol/kg) or 0.15 M NaCl (n=4/group). Additionally, fasted mice were injected ip with obestatin (1 micromol/kg) or urocortin 1 (2 nmol/kg) 15 min before food presentation. No effect on FI was observed after obestatin administration during the light and dark phase under both metabolic conditions while CCK8S reduced FI irrespectively of the conditions. The number of Fos positive neurons was not modified by obestatin while CCK8S increased Fos expression in selective brain nuclei. Obestatin did not influence the refeeding response to a fast in mice, while urocortin was effective. Therefore, peripheral obestatin has no effect on FI under various experimental conditions and did not induce Fos in relevant central neuronal circuitries modulating feeding in rodents.


Protection of immunosuppressed mice against infection with pseudomonas aeruginosa by monoclonal antibodies to outer membrane protein OprI.
Two monoclonal antibodies (mAbs) designated 2A1 and 6A4 which had been shown to be directed against Pseudomonas aeruginosa outer membrane lipoprotein I were tested in cyclophosphamide treated mice for their protective ability against P. aeruginosa infection. Pretreatment of mice with either 2 mg of 2A1 or 4 mg of 6A4 in combination with lethally irradiated human leucocytes reduced the mortality after subsequent subcutaneous injection of 100 living P. aeruginosa organisms to 50% of the controls. Without leucocytes only mAb 2A1 (isotype IgG2b) showed protection (47%). Irradiated leucocytes alone without mAb did not protect the mice significantly from fatal P. aeruginosa infection.


WHO Working Group on revision of the Manual of Laboratory Methods for Testing DTP Vaccines-Report of two meetings held on 20-21 July 2006 and 28-30 March 2007, Geneva, Switzerland.
This report reflects the discussion and conclusions of a WHO group of experts from National Regulatory Authorities (NRAs), National Control Laboratories (NCLs), vaccine industries and other relevant institutions involved in standardization and control of diphtheria, tetanus and pertussis vaccines (DTP), held on 20-21 July 2006 and 28-30 March 2007, in Geneva Switzerland for the revision of WHO Manual for quality control of DTP vaccines. Taking into account recent developments and standardization in quality control methods and the revision of WHO recommendations for D, T, P vaccines, and a need for updating the manual has been recognized. In these two meetings the current situation of quality control methods in terms of potency, safety and identity tests for DTP vaccines and statistical analysis of data were reviewed. Based on the WHO recommendations and recent validation of testing methods, the content of current manual were reviewed and discussed. The group agreed that the principles to be observed in selecting methods included identifying those critical for assuring safety, efficacy and quality and which were consistent with WHO recommendations/requirements. Methods that were well recognized but not yet included in current Recommendations should be taken into account. These would include in vivo and/or in vitro methods for determining potency, safety testing and identity. The statistical analysis of the data should be revised and updated. It was noted that the mouse based assays for toxoid potency were still quite widely used and it was desirable to establish appropriate standards for these to enable the results to be related to the standard guinea pig assays. The working group was met again to review the first drafts and to input further suggestions or amendments to the contributions of the drafting groups. The revised manual was to be finalized and published by WHO.


Kinetics of plasma corticosteroids during metyrapone infusion in man.
The effect of metyrapone on plasma kinetics of cortisol (F), 11-deoxycortisol (S), corticosterone (B), 11-deoxycorticosterone (DOC), aldosterone (Aldo) and 17 hydroxyprogesterone (17 OHP) was studied in ten normal prepubertal children intravenously infused with the drug (1 gm/1.73 m2 body surface area in 4 h). Similar falls in F, B and Aldo were observed during the first hour expressing similar sensitivity of the 11 hydroxylases to the drug. The effect of the compensatory increase of endogenous ACTH could be detected after 1 hour by the rise in 17 OHP. However the concentrations of the 11-hydroxy compounds, F, B and Aldo, remained low throughout the infusion period. From the second hour onwards, facilitation of the synthesis of DOC was noticed. After the end of infusion, the levels of the precursors decreased and F and B were restored to initial values. Aldo remained low probably due to the inhibitory effect of the elevated DOC on the renin-angiotensin system.


Thiopental sodium attenuates hypoxia/reoxygenation-induced injury in osteoblasts by modulating AKT signaling.
Thiopental sodium (TPTS) is a barbiturate general anesthetic, while its effects on hypoxia/reoxygenation (H/R)-induced injury are still unclear. This study aimed to investigate whether TPTS exerts protective effects against the H/R-induced osteoblast cell injury and explore the underlying mechanisms. Osteoblast cell injury model was induced by the H/R condition, which was treated with or without TPTS. Cell viability and lactate dehydrogenase (LDH) release were determined by the corresponding commercial kits. The levels of oxidative stress were determined in the experimental groups. Cell apoptosis and Caspase-3 activities were determined by propidium iodide staining and substrate-based assay, respectively. Western blotting and qRT-PCR were performed to measure the mRNA and protein levels, respectively. Treatment with TPTS was able to increase cell viability and reduce LDH release in H/R-induced osteoblasts. Additionally, TPTS regulated oxidative stress in H/R-induced osteoblasts by suppressing malondialdehyde (MDA) and reactive oxygen species (ROS) as well as boosting superoxide dismutase (SOD). TPTS was able to suppress cell apoptosis by suppressing Caspase-3 activity and cleavage. TPTS exerted protective effects against cell injury and apoptosis induced by the H/R conditions, which were associated with its regulation of Akt signaling. Moreover, TPTS induced osteoblast differentiation under the H/R condition. In summary, TPTS attenuates H/R-induced injury in osteoblasts by regulating AKT signaling.


Characterization of the complete mitochondrial genome of Amblyomma ovale, comparative analyses and phylogenetic considerations.
In this study, we sequenced two complete mitochondrial genomes of Amblyomma ovale, a tick of public health importance. Sequencing two distinct individuals, the resulting mitochondrial genomes were 14,756 and 14,760 bp in length and maintained the same gene order previously reported in Amblyomma. These were combined with RNA-seq derived mitochondrial sequences from three additional species, Amblyomma aureolatum, Amblyomma maculatum, and Amblyomma moreliae, to carry out mitogenome comparative and evolutionary analyses against all previously published tick mitochondrial genomes. We described a derivative genome rearrangement that isolates Ixodes from the remaining Ixodidae and consists of both a reverse translocation as well as an event of Tandem Duplication Random Loss. Genetic distance analyses indicated that cox2, nd1, nd5, and 16S are good candidates for future population studies in A. ovale. The phylogenetic analyses corroborated the utility of complete mitochondrial genomes as phylogenetic markers within the group. This study further supplements the genome information available for Amblyomma and facilitates future evolutionary and population genetic studies within the genus.


Mycotoxins aptasensing: From molecular docking to electrochemical detection of deoxynivalenol.
This work proposes a voltammetric aptasensor to detect deoxynivalenol (DON) mycotoxin. The development steps of the aptasensor were partnered for the first time to a computational study to gain insights onto the molecular mechanisms involved into the interaction between a thiol-tethered DNA aptamer (80mer-SH) and DON. The exploited docking study allowed to find the binding region of the oligonucleotide sequence and to determine DON preferred orientation. A biotinylated oligonucleotide sequence (20mer-BIO) complementary to the aptamer was chosen to carry out a competitive format. Graphite screen-printed electrodes (GSPEs) were electrochemically modified with polyaniline and gold nanoparticles (AuNPs@PANI) by means of cyclic voltammetry (CV) and worked as a scaffold for the immobilization of the DNA aptamer. Solutions containing increasing concentrations of DON and a fixed amount of 20mer-BIO were dropped onto the aptasensor surface: the resulting hybrids were labeled with an alkaline phosphatase (ALP) conjugate to hydrolyze 1-naphthyl phosphate (1-NPP) substrate into 1-naphthol product, detected by differential pulse voltammetry (DPV). According to its competitive format, the aptasensor response was signal-off in the range 5.0-30.0 ng·mL-1 DON. A detection limit of 3.2 ng·mL-1 was achieved within a 1-hour detection time. Preliminary experiments on maize flour samples spiked with DON yielded good recovery values.


Inhibitory effect of L-glutamine on gastric irritation and back diffusion of gastric acid in response to aspirin in the rat.
L-Glutamine given orally at 750 mg/kg significantly reduced the formation of gastric mucosal lesions induced by aspirin at 100 mg/kg at 1, 3, and 7 hours in pylorus-ligated rats. L-Glutamine markedly inhibited the loss of H+ and a corresponding increment of Na+ through the aspirin-damaged mucosal barrier, which suggests inhibition of acid back diffusion as the mechanism of action. In vagotomized rats, the diffusion of H+ from the instilled acid solution into the gastric mucosa and outflux of Na+ from the mucosa into the lumen was so strong that aspirin could not show any acceleration of a back diffusion of H+ in contrast to the aspirin-free group. However, L-Glutamine given with or without aspirin inhibited the group. However, L-Glutamine given with or without aspirin inhibited the back diffusion of H+ instilled into the vagotomized rat stomach.


Polydatin suppresses proliferation and metastasis of non-small cell lung cancer cells by inhibiting NLRP3 inflammasome activation via NF-κB pathway.
Non-small cell lung cancer (NSCLC) is the leading cause of cancer death. Particularly, inflammation is crucial for the progression of NSCLC. In the past few decades, the anti-tumor effects of some traditional Chinese medicinal herbs have caused much attention. This study was designed to investigate the effects of polydatin in the progression of NSCLC. CCK-8 assay, wound healing assay and western blot assay were used to assess the anti-cancer property. Consequently, we showed that polydatin inhibited proliferation and migration of NSCLC cells (A549 and H1299 cells) in a dose-dependent manner. In addition, polydatin suppressed the expression of NLRP3, ASC and pro-caspase-1in NSCLC cells. Activation of NLRP3 inflammasome counteracted the inhibitory effect of polydatin on proliferation and migration of NSCLC cells, suggesting that polydatin suppressed progression of NSCLC through inhibiting NLRP3 inflammasome activation. Furthermore, polydatin was found to down-regulate relative expression of phosphor-NF-κB p65 and activation of NF-κB pathway by TNF-α also abolished the inhibitory effect of polydatin on proliferation and migration of NSCLC cells. In conclusion, our data showed that polydatin acted as an anti-tumor agent to suppress proliferation and metastasis of NSCLC cells. The anti-tumor effect of polydatin was possibly related to the inhibition of NLRP3 inflammation via the NF-κB pathway. Our finding suggested that polydatin might be a potential therapeutic candidate in the treatment of NSCLC.


Constructing hollow ZIF-8/CDs@MIPs fluorescent sensor from Osmanthus leaves to specifically recognize bovine hemoglobin.
To develop non-toxic, highly efficient and selective fluorescence sensors was a significance research. In this work, a novel hollow fluorescence sensor was designed with biomass carbon dots (CDs), ZIF-8 and molecularly imprinted polymers (MIPs) via aqueous polymerization. The results demonstrated such unique structure fluorescence sensor exhibited fast response time, excellent stability and highly selectively towards bovine hemoglobin (BHb). Even in a complex environment, the hollow fluorescence sensor (H-ZIF-8/CDs@MIPs) still has a good recognition effect on BHb. Under an optimized condition, the hollow fluorescence sensor was quenched linearly with BHb concentration in the range of 0.058-4.5 μM with the detection limit of 15.6 nM. In addition, a possible quenching mechanism of hollow fluorescence towards BHb was confirmed resonance energy transfer (FRET). In the actual application process, the hollow fluorescence sensor showed a better detection performance towards BHb with the recoveries ranged of 98.6-101.1 %. This work provided a strategy to design green and unique hollow fluorescence sensor for practical application.


Nonviral and viral gene transfer to the kidney in the context of transplantation.
Local modulation of the immune response through genetic manipulation of the graft is an attractive novel approach to overcome the toxicity of immunosuppressive therapy to prevent acute graft rejection. We have previously reported that the cationic polymer polyethylenimine 25k (PEI 25k) transduced reporter genes when injected into the renal artery, but with a low transfection efficiency. Here we compare the risk/benefit profiles of such a nonviral versus a viral technique of gene transfer to the kidney in the context of renal transplantation. Donor kidneys from Lewis rats were perfused in a closed circuit with an artificial cell-free medium containing PEI 25k complexed to an expression vector coding for the beta-galactosidase (beta-gal) gene and subsequently transplanted in a syngeneic animal. In a second set of experiments, donor kidneys were injected or perfused with a replication-deficient adenovirus encoding the beta-gal gene (AdCMV. beta-gal; 1 x 10(9) plaque-forming units) before transplantation. Perfusion of the kidney with PEI 25k/DNA complexes resulted in large areas of hypoperfusion characterized by injured glomeruli and tubuli, capillary thrombosis and accumulation of C3 in glomerular capillaries. Reperfusion of the kidney was achieved by lowering the PEI 25k/DNA ratio, but no detectable transfection was observed. In animals receiving adenovirus, the beta-gal activity increased with time and was localized mainly in proximal and distal tubular cells, as documented by beta-gal histochemistry and in situ hybridization. A significantly increased expression of beta-gal was achieved by perfusion of the kidney with AdCMV.beta-gal before transplantation, beta-gal staining mainly localizing in proximal and distal tubular cells. Unlike nonviral methods of gene delivery, adenovirus-mediated gene transfer to the kidney offers exciting perspectives for the development of molecular medicine in the field of organ transplantation.


Coverage of mass drug administration for elimination of lymphatic filariasis in urban Nagpur, Central India: A mixed method study.
Mass Drug Administration (MDA) coverage remains an important indicator in elimination of Lymphatic Filariasis (LF), especially in context of recent changes in programme strategies in India, such as incorporation of Ivermectin and involvement of urban Accredited Social Health Activists (ASHAs). This study aimed at assessing the coverage and compliance with MDA of Filariasis as well as exploring perspective of beneficiaries for non-consumption in selected slum area of Nagpur city. Mixed-method study design comprising of quantitative assessment of MDA coverage, followed by qualitative method to explore the reasons of non-compliance was used in selected slum areas of Nagpur city. Using cluster sampling, 240 households were selected and house-to-house visits were made to interview the eligible participants. In-depth interviews were conducted among selected non-compliant participants. Multivariable logistic regression analysis to identify the factors associated with non-consumption. Thematic analysis was done to obtain the reasons of non-consumption as perceived by the beneficiaries. Among the 1096 individuals studied, distribution and consumption coverage were 55.2% and 48.5%, respectively. Effective supervised consumption was further low (28.9%). Coverage compliance gap (CCG) was 12.1%. Male sex and younger age (2-5 years) were significant socio-demographic determinants of non-consumption. No repeat visit to houses left in first round, fear of side effects, pill burden, poor understanding about the need were important reasons as revealed by qualitative inquiry. Effective pre-campaign awareness, incorporation of context specific drug delivery strategies and strengthening monitoring system are essential for successful MDA implementation.


Chemotherapy agents reduce protein synthesis and ribosomal capacity in myotubes independent of oxidative stress.
Chemotherapeutic agents (CAs) are first-line antineoplastic treatments against a wide variety of cancers. Despite their effectiveness in halting tumor progression, side effects associated with CAs promote muscle loss by incompletely understood mechanisms. To address this problem, we first identified how oxidative stress impairs protein synthesis in C2C12 myotubes. Transient elevations in reactive oxygen species (ROS) resulted in protein synthesis deficits and reduced ribosomal (r)RNA levels. Oxidative stress did not reduce rRNA gene (rDNA) transcription, but it caused an increase in rRNA and protein oxidation. To determine whether CAs affect protein synthesis independent of oxidative stress, we exposed myotubes to Paclitaxel (PTX), Doxorubicin (DXR), or Marizomib (Mzb) at doses that did result in elevated ROS levels (sub-ROS). Exposure to CAs reduced protein synthesis and rRNA levels, but unlike oxidative stress, sub-ROS exposures impaired rDNA transcription. These results indicate that although oxidative stress disrupts protein synthesis by compromising ribosomal quantity and quality, CAs at sub-ROS doses compromise protein synthesis and ribosomal capacity, at least in part, by reducing rDNA transcription. Therefore, CAs negatively impact protein synthesis by causing oxidative stress in addition to directly reducing the ribosomal capacity of myotubes in a ROS-independent manner.


Open status of pig-breeding farms is associated with slightly higher seroprevalence of F18+ Escherichia coli in northern Belgium.
F18+ enterotoxigenic and verotoxigenic Escherichia coli (E. coli) are widespread and induce post-weaning diarrhoea and oedema disease, respectively. No commercial vaccine exists. Therefore, the F18 herd seroprevalence of 178 randomly selected pig-breeding farms in northern Belgium was evaluated by determining F18-specific IgG antibody titres in pooled sera of sows per breeding farm. This survey reported the first data concerning F18 herd seroprevalence and showed that F18+ E. coli seroprevalence was common in northern Belgium. Significantly more open pig-breeding farms were F18 seropositive (96.4%) compared to closed (88.8%). Furthermore, no clear association was determined between the F18 herd seroprevalence and the number of sows per breeding farm (rsp = 0.19), or with the number of fattening pigs per breeding farm (rsp = 0.17).


Screening of renal clear cell carcinoma prognostic marker genes based on TCGA and GTEx chip data and construction of transcription factor-related regulatory networks.
This study aimed to identify prognostic marker genes for renal clear cell carcinoma (RCCC) and construct a regulatory network of transcription factors and prognostic marker genes. Three hundred eighty-six genes were significantly differentially expressed in RCCC, with functional enrichment analysis suggesting a relationship between these genes and kidney function and development. Cox and Lasso regression analyses revealed 10 prognostic marker genes (RNASET2, MSC, DPEP1, FGF1, ATP1A1, CLDN10, PLG, SLC44A1, PCSK1N, and LGI4) that accurately predicted RCCC patient prognosis. Upstream transcription factors of these genes were also identified, and in vitro experiments suggested that ATP1A1 may play a key role in RCCC patient prognosis. The findings of this study provide important insights into the molecular mechanisms of RCCC and may have implications for personalized treatment strategies.


Serum long-chain omega-3 polyunsaturated Fatty acids and future blood pressure in an ageing population.
To investigate the associations of serum long-chain omega-3 polyunsaturated fatty acids (PUFA) and hair mercury with future blood pressure in an ageing population. Prospective study with baseline measurements in 1998-2001 and follow-up measurements in 2005-2008. The linear relationships (β) of baseline serum fatty acids and hair mercury with future systolic and diastolic blood pressure and pulse pressure were analyzed with multiple linear regression models, using log-transformed values. 181 men and 200 women aged 53-73 y from the Kuopio Ischemic Heart Disease Risk Factor Study (KIHD) population in Eastern Finland, who were free of cardiovascular disease, diabetes or hypertension at baseline. Total serum esterified and nonesterified fatty acids and pubic hair mercury were used as markers for exposure. Anthropometric and other lifestyle and health-related data were collected. The mean serum concentrations were 1.67% (SD 0.92) for eicosapentaenoic acid (EPA), 0.79% (SD 0.16) for docosapentaenoic acid (DPA) and 2.78 (SD 0.92) for docosahexaenoic acid (DHA), of all serum fatty acids. The mean hair mercury concentration was 1.5 µg/g (SD 1.6). We did not find statistically significant associations between the baseline serum long-chain omega-3 PUFA concentrations or hair mercury content and future blood pressure. Hair mercury did not modify the associations with the long-chain omega-3 PUFAs, either. Higher serum long-chain omega-3 PUFA concentration, a biomarker of fish or fish oil consumption, may not have an impact on future blood pressure in an ageing population.


Intraocular tetrodotoxin in goldfish hinders optic nerve regeneration.
Repeated intraocular injection of tetrodotoxin (TTX) was used to produce a maintained block of neural activity in goldfish optic axons which were regenerating following a crush of the optic nerve. The recovery of visual function was delayed in the TTX-treated fish, as demonstrated by delays in the return of the startle reaction to sudden illumination, the dorsal light reflex and food pellet localization. A single injection of TTX at the time of optic nerve crush delayed recovery of the startle reaction but not of food localization. There was no loss of ganglion cells in the TTX-treated animals, but the number of regenerated axons detectable by light microscopy was reduced. Also, axonal transport of radioactively labeled protein in some of the regenerating axons showed a deficit at 21-28 days after the lesion, i.e., during innervation of the tectum. Incorporation of labeled amino acid into protein in the retinal ganglion cells was depressed during the same period, but both the transport and incorporation had returned to normal by 36 days after the lesion. These results, together with the results of the accompanying electrophysiological analysis of the effects of TTX58,59, suggest that TTX treatment interferes with two separate events in regeneration, one occurring soon after the nerve lesion and the other during innervation of the tectum. During at least the first of these events the effect of TTX treatment may be to reduce the number of size of the regenerating axon branches. We propose that the TTX effect may be mediated by a reduction in the axonal transport of certain materials, including gangliosides, nucleosides, or proteins specifically involved in regeneration.


Primary intradural extramedullary spinal Burkitt's lymphoma mimicking a nerve sheath tumor: a case report.
Spinal involvement in lymphomas is often associated with advanced disease. Primary spinal non-Hodgkin's lymphoma is a rare entity. A 47-year-old male presented with a history of neck pain followed by progressive quadriparesis and bowel bladder involvement over a 5-month period. The magnetic resonance imaging was suggestive of an intradural extramedullary lesion at the C1-C2 vertebra level. A surgical excision was done and the histopathology revealed atypical lymphoid cells, which are immunopositive for CD45, CD20, MUM-1, and BCL6, while negative for BCL2, EBV (LMP-1 and CISH), Cyclin D1 and confirmed the diagnosis of Burkitt's lymphoma. The patient received chemotherapy in the form of CODOX-M/IVAC (cyclophosphamide, vincristine, doxorubicin, high-dose methotrexate/ifosfamide, etoposide, high-dose cytarabine) regimen. Primary spinal intradural extramedullary Burkitt's lymphoma is a rare diagnosis that may often be difficult to differentiate radiologically from other causes of intradural extramedullary lesions. A thorough histological examination is warranted in such cases.


Evolution of TNM Classification for Clinical Staging of Oral Cancer: The Past, Present and the Future.
The AJCC (American Joint Committee on Cancer) Cancer Staging Manual, 1st Edition, was published in 1977 which focused on the TNM classification and staging of cancer to allow easy communication, formulation of a treatment plan and predict the prognosis, among the medical fraternity. Ever since the beginning, various modifications of the classification were introduced and released by the joint collaboration of AJCC and UICC (International Union Against Cancer) in various editions of cancer staging manuals. The present review article was kept focused onto the changes introduced in the clinical staging of cancers of oral cavity. These changes came a long way since 1st edition 1944, to the eighth edition which was published in 2017. This article is a critical review on the past and present perspectives of the TNM classification of the oral cavity that were addressed and changed, adding a light on the future trends or necessary inclusions that would formulate a much easily acceptable and useful classification system.


Objective-subjective disparity in cancer-related cognitive impairment: does the use of change measures help reconcile the difference?
Studies to date have found little correlation between subjective and objective measures of cognitive function in cancer patients, making it difficult to interpret the significance of their cognitive complaints. The purpose of this study was to determine if a stronger correlation would be obtained using measures of cognitive change rather than static scores. Sixty women with early stage breast cancer underwent repeated cognitive assessment over the course of chemotherapy with a neuropsychological test battery (objective measure) and with the FACT-Cog (subjective measure). Their results were compared to 60 healthy women matched on age and education and assessed at similar intervals. We used multilevel modeling, with FACT-Cog as the dependent measure and ordinary least squares slopes of a neuropsychological summary score as the independent variable, to evaluate the co-variation between the subjective and objective measures over time RESULTS: Measures of both objective and subjective cognitive function declined over the course of chemotherapy in the breast cancer patients but there was no significant relationship between them, even when using change measures. Change in objective cognitive function was not related to change in anxiety or fatigue scores but the decline in perceived cognitive function was associated with greater anxiety and fatigue. The discrepancy in objective and subjective measures of cognition in breast cancer patients cannot be accounted for in terms of a failure to use change measures. Although the results are negative, we contend that this is the more appropriate methodology for analyzing cancer-related changes in cognition.


Retinoic acid receptors and tissue-transglutaminase mediate short-term effect of retinoic acid on migration and invasion of neuroblastoma SH-SY5Y cells.
Long-term treatment with all trans-retinoic acid (RA) induces neuronal differentiation and apoptosis. However, the effect of short-term RA treatment on cell proliferation, migration and invasion of neuroblastoma cell lines (SH-SY5Y and IMR-32) remains unclear. RA induces expression of tissue-transglutaminase (TGase) and promotes migration and invasion after 24 h of treatment in SH-SY5Y cells, but not in IMR-32 cells. RA receptor (RAR) agonist (4-(E-2-[5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl]-1-propenyl) benzoic acid) and RAR/retinoid X receptor (RXR) agonist (9-cis-RA) promote expression of TGase, migration and invasion of SH-SY5Y cells, while RXR agonist has no significant effect. RAR antagonist blocks RA effect on migration and invasion, indicating that RAR receptors are required. Retinoid receptors are expressed and activated by RA in both cell lines. However, only transient activation of RAR is observed in IMR-32 cells. These findings suggest that different responses observed in SH-SY5Y and IMR-32 cells could be due to differential activation of retinoid receptors. Overexpression of TGase has no effect on migration or invasion, while overexpression of antisense TGase blocks RA-induced migration and invasion, indicating that other molecules along with TGase mediate RA effects. In addition to the long-term effects of RA that are coupled with cell differentiation, short-term effects involve migration and invasion of neuroblastoma SH-SY5Y cells.


Ethnicity and weight status affect the accuracy of proxy indices of insulin sensitivity.
This study tested the hypotheses that correlations between direct measures of insulin sensitivity and proxy indices of insulin sensitivity derived from fasting values, (i) would not be affected by ethnicity, and (ii) would be stronger in overweight vs. weight-reduced states. We further hypothesized that associations between proxy indices and fat distribution would be similar to those between directly measured insulin sensitivity and fat distribution. Testing was performed in weight-stable conditions in 59 African-American (AA) and 62 white-American (WA) overweight, premenopausal women before and after a weight loss intervention. Subjects were retested 1 year following weight loss. Proxy indices were correlated against the insulin sensitivity index S(I) determined via minimal modeling. Fat distribution was assessed using computed tomography. Correlations between Si and proxy indices were consistently stronger among overweight women (r = 0.44-0.52) vs. weight-reduced women (r = 0.18-0.32), and among AA (r = 0.49-0.56, baseline; 0.24-0.36, weight-reduced) vs. WA (r = 0.38-0.46, baseline; 0.19-0.31, weight-reduced). Among subjects who regained >3 kg after 1 year, correlations between S(I) and proxy indices were similar to those observed at baseline, whereas correlations were weak among women who maintained their reduced body weight. S(I) and all proxy indices were similarly correlated with intra-abdominal adipose tissue (IAAT) at baseline, but not after weight loss. In conclusion, correlations between S(I) and proxy indices were affected by both ethnicity and weight status. If proxy indices are used in multiethnic populations, or in populations including both lean and overweight/obese subjects, data should be interpreted with caution.


Cytokinin (6-benzylaminopurine) elevates lignification and the expression of genes involved in lignin biosynthesis of carrot.
Carrot is a root crop consumed worldwide and has great nutritional qualities. It is considered as one of the ten most important vegetable crops. Cytokinins are an essential class of the plant hormones that regulate many processes of plant growth. Till now, the effects of cytokinin, BAP, on lignin biosynthesis and related gene expression profiles in carrot taproot is unclear. In order to investigate the effect of applied BAP on lignin-related gene expression profiles, lignin accumulation, anatomical structures, and morphological characters in carrot taproots. Carrot roots were treated with different concentrations of BAP (0, 10, 20, and 30 mg L-1). The results showed that the application of BAP significantly increased plant length, shoot fresh weight, root fresh weight, and taproot diameter. In addition, BAP at 20 mg L-1 or 30 mg L-1 enhanced the average number of petioles. BAP treatment led to increased number and width of xylem vessels. The parenchyma cell numbers of pith were significantly induced in taproots treated with the BAP at a concentration of 30 mg L-1. BAP significantly upregulated most of the expression levels of lignin biosynthesis genes, caused elevated lignin accumulation in carrot taproots. Our results indicate that BAP may play important roles in growth development and lignification in carrot taproots. Our results provide a valuable database for more studies, which may focus on the regulation of root lignification via controlling cytokinin levels in carrot taproots.


Bovine chondrocyte behaviour in three-dimensional type I collagen gel in terms of gel contraction, proliferation and gene expression.
This study evaluated the in vitro behaviour of bovine chondrocytes seeded in collagen gels, promising recently reported scaffolds for the treatment of full-thickness cartilage defects. To determine how chondrocytes respond to a collagen gel environment, 2 x 10(6) chondrocytes isolated from fetal, calf and adult bovine cartilage were seeded within type I collagen gels and grown for 12 days in both attached and floating (detached from the culture dish after polymerisation) conditions. Monolayer cultures were performed in parallel. All chondrocytes contracted floating gels to 55% of the initial size, by day 12. Contraction was dependent on initial cell density and inhibited by the presence of dihydrocytochalasin B as previously observed with fibroblasts. Gene expression was determined using conventional and real-time PCR. The chondrocyte phenotype was better maintained in floating gels compared to attached gels and monolayers. This was demonstrated by comparing the ratio of COL2A1/ COL1A2 mRNA and also of alpha10/alpha11 integrin mRNA. A strong up-regulation of MMP13 expression was measured at day 12 in floating gels. The composition of cartilage-like tissue obtained by growing chondrocytes in a collagen gel varied depending on the floating or attached conditions and initial cell density. It is thus important to consider these parameters when using this culture system in order to prepare a well-defined implant for cartilage repair.


The clastogenicity of morphine sulfate in vivo.
An opioid analgesic, morphine, and an opioid peptide, beta-endorphin, have been shown to induce chromosome damage, as indicated by an increased frequency of micronucleated lymphocytes, following acute administration to mice. The genotoxic response is opioid receptor-mediated and is abolished in adrenalectomized animals. Further, plasma from morphine-treated animals also induces micronuclei formation in naive lymphocytes in vitro; this response is blocked by inclusion the steroid antagonist RU 486 in the incubation mixture. In addition to the steroid-mediated production of chromosome damage, morphine acts directly on lymphocytes to enhance the clastogenicity of acutely administered cyclophosphamide in manner consistent with depressed DNA repair capacity.


Systems Genetics of Liver Fibrosis.
This systems genetics analysis comprises quantitative measurements of hepatic fibrogenesis in mouse models and mapping of quantitative traits in mouse genetic reference populations. It is part of a large mapping project of fibrogenic genes including the analyses of experimental crosses from different inbred mouse strains. Extensive quantitative trait loci (QTL) mapping of fibrosis phenotypes and liver expression profiling in combination with in silico mapping facilitated the identification of QTL regions and underlying candidate genes that confer fibrosis susceptibility also in humans. Moreover, the approach led to the identification of interacting QTLs and gene networks in liver fibrosis, providing a key experimental platform for the development of novel, more precise therapeutic interventions. Here, we provide a use case for the application of different analysis tools and the integration of multiple datasets determined in F2 intercrosses and BXD recombinant inbred lines to identify, finemap and affirm fibrosis susceptibility loci.


Possible role of platelet-activating factor in the in vivo expression of tissue factor in neutrophils.
Recently, we demonstrated that neutrophils express tissue factor (TF) in a model of acute obstructive cholangitis (AOC). However, the regulation of TF expression was not clear. In this study, we clarified the role of platelet-activating factor (PAF) in TF expression in neutrophils. In a model of AOC, intravenous PAF antagonist, (SM-12502, 200 mg/kg) was administered 5 min before sepsis was induced. Normal saline was given as a control. Coagulation parameters and TF activity were monitored for 6 h. Thereafter, the liver was harvested for histological examination. The percentage of neutrophils which stained positive for TF was significantly reduced by SM-12502 (74.9 +/- 19.3% vs 96.3 +/- 2.8%) (P < 0.01). The number of leukocytes infiltrating the liver was also significantly reduced. Coagulation abnormalities, TF activity, and focal necrosis of the hepatocytes were reduced by SM-12502. SM-12502 inhibits TF expression in neutrophils which have infiltrated the liver sinusoids, reducing the subsequent infiltration of leukocytes. These results suggest that PAF plays an important role in the expression of TF in neutrophils in vivo.


A description of dry oral forms of medication administration modifications by nurses in home settings.
Modifying the galenic of dry oral forms (DOF) to be administered to patients with swallowing or behavioral disorders is frequent in long term care homes. The objective was to investigate the practice of modifying DOF galenic by nurses in home settings (NHS). A 14-question electronic survey was distributed to 1977 NHS. Almost 3/4 of respondents reported crushing tablets or opening capsules, with 37% on a daily basis. Approximately 22% did not inquire about the feasibility of modifying galenic DOF beforehand. Nearly 75% of NHS were aware about the risk of ineffectiveness after crushing tablets or opening capsules. However, only 14% mentioned the risk of overdosing associated with this practice. More than 60% of NHS never wore protective equipment when changing dosage form. Changing the dosage form of DOF is a widespread practice in home settings. Our present work urges the need to improve collaboration between prescribing physicians, nurses and pharmacists.


Cost-effectiveness of preoperative magnetic resonance imaging to optimize surgery in ductal carcinoma in situ of the breast.
Complete surgical excision is the main factor for successful breast-conserving surgery in patients with ductal carcinoma in situ (DCIS) of the breast. Preoperative magnetic resonance imaging (MRI) may allow surgery optimization in this indication. From an economic standpoint, systematic preoperative MRI is associated with an extra cost, which may be offset by a decrease in the number of re-interventions. We performed an economic evaluation alongside IRCIS randomised controlled trial (NCT01112254) to determine whether systematic preoperative MRI in DCIS is a cost-effective strategy. 360 patients were included in IRCIS trial. Costs were assessed from the French national health insurance perspective. Resource use was prospectively collected during a 6-month period after randomisation. We estimated the mean cost per averted re-intervention. Despite extra costs due to MRI and additional biopsies, difference in total costs between arms was not statistically significant (mean cost of €9980 in MRI arm and €9682 in no MRI arm, cost difference: €298 [CI95% : -470; 1063]). There was a non-significant decrease in the rate of re-hospitalisations for positive or close margins (20% in MRI arm versus 27% in No MRI arm, difference -7% [CI95% : -17; 3]). At a willingness to pay of €500 to avert a re-intervention, the probability that MRI strategy is cost-effective was 93%. Systematic preoperative MRI in patients with DCIS of the breast may be a cost-effective strategy. However, the modest clinical benefit associated with such a strategy limits the interest for this procedure in routine practice given the current MRI techniques.


The effect of prior adult experience on components of habitat preference in the apple maggot fly (Rhagoletis pomonella).
Numerous authors have suggested that genetic subdivision within a population in a heterogeneous environment is more likely if individuals tend, through prior experience, to breed in the same habitat in which they developed. Under semi-field conditions we demonstrate that prior adult experience alters habitat preference in the apple maggot fly, Rhagoletis pomonella (Tephritidae), a frugivorous parasitic insect thought to have undergone sympatric divergence in host use in historical times. Females exposed to a particular host fruit species - apple (Malus pumila) or hawthorn (Crataegus mollis) - in a field cage oviposited at a higher rate in test fruit of that species than did inexperienced females or females exposed to the other species. Females exposed to a particular host fruit species also tended to remain longer in test trees harboring fruit of that species than did inexperienced females or females exposed to the other species. Prior adult experience thus alters two components of habitat preference in the apple maggot fly: oviposition preference and habitat fidelity. We discuss how these effects of experience on habitat preference should increase the likelihood that individuals mate assortatively and may further increase the likelihood that apple maggot populations become genetically subdivided.


Distribution, sources and transport of polycyclic aromatic hydrocarbons (PAHs) in karst spring systems from Western Hubei, Central China.
Karst groundwater is an important water resource but it is vulnerable to contaminants, due to the distinctive geological features of abundant transmissive fractures and conduits in the karst area which connect the surface to the underground systems. Anthropogenic activity-derived polycyclic aromatic hydrocarbons (PAHs) on the surface environment could enter groundwater easily and rapidly and threaten water security in karst areas. Samples in the multimedia environment from 10 specific karst spring systems from Western Hubei of Central China were collected to analyze 16 priority PAHs and to investigate their transport in these karst spring systems. The total concentrations of PAHs in the soil, river water, river sediments, spring water, and spring sediments ranged between 6.04 and 67.7 ng g-1, 4.56 and 11.4 ng L-1, 29.9 and 1041 ng g-1, 4.09 and 222 ng L-1, and 5.88 and 83.0 ng g-1, respectively. Levels of PAHs in this area were relatively low when compared to other karst areas. Proportions of low-molecular-weight (LMW)-PAHs in the water, sediments and soil (average 58.2-78.8%) were much higher than those of high-molecular-weight (HMW)-PAHs. The proportion of LMW-PAHs in the sediments (especially in river sediments) was higher than that in the soil. Characteristic ratio analysis and principal component analysis showed that PAHs were from high-temperature combustion of the mixture of coal and biomass, and vehicle emission, where coal and biomass combustion were the dominant sources. Significant correlations of PAH compositions in different media of karst spring systems were observed, especially in the Yuquangdong (YQD)-Migongquan (MGQ), Jiuzhenziquan (JZZQ), Xianyudong (XYD) and Fengdong (FD) karst spring systems, indicating the rapid PAH transport from the recharge area soil to the discharge area of spring water and sediments.


Diet Quality in Early Care and Education Centers: A Comparison of Menu, Served, and Consumed Lunch Measures.
Assess if diet quality (DQ) differs among 3 lunch measures commonly used to assess DQ in early care and education (ECE) centers, which include what is listed on the menu, served, and consumed. This quantitative observational study measured DQ of food and beverage items listed on the menu, what children were served, and what they consumed during one lunch meal. Alabama ECE centers. ECE centers (n = 28) with attendance of ≥10 children ages 3 to 5 years and provided lunch daily. DQ measured through Healthy Eating Index-2015 (HEI) scores, which were calculated for the menu, served, and consumed lunch measures. Menu, served, and consumed HEI scores compared using either Kruskal-Wallis or ANOVA. Dunn Bonferroni or Duncan post hoc tests identified which group differed. Total HEI scores were significantly higher among menus, compared with served and consumed (P = .001). Served and consumed total HEI scores were not different. Menu DQ differed from both the served and consumed lunch measures. These findings support the need for nutrition education on the importance of menu adherence in ECE centers.


Microscopic spermatic cord denervation for chronic orchialgia/chronic scrotal content pain: operative outcomes and predictors of failure.
To describe our institutional outcomes with microscopic spermatic cord denervation (MSCD) for chronic scrotal content paint (CSCP) and identify predictors of treatment failure. Retrospective chart review was performed to identify all MSCD performed by two surgeons at a single institution from 2010-2019. Patient demographic data and operative outcomes were collected. Patients were excluded from analysis if no post-operative follow up was available. Success was defined as complete resolution of bothersome pain. Multivariable logistic regression was utilized to identify predictors of treatment failure. During the study period, 101 patients were identified in which 113 MSCD procedures were performed. Final analysis included 103 procedures across 93 patients. Mean age was 41.8 years (SD 13.2), mean BMI was 29.2 kg/m2 (SD 5.96) and median months of pain preceding surgery were 24 (range, 3-300 months). Overall, 75/103 (73%) MSCD were successful. Of the failures, 5 patients had recurrence of pain greater than 6 months after surgery. Only the presence of pelvic floor muscle spasm (PFMS) independently predicted MSCD failure (OR 3.95, P=0.02). 9 of 19 (47%) patients with PFMS experienced treatment failure, while 19 of 84 (23%) without PFMS experienced failure. MSCD offers a therapeutic option for patients with refractory CSCP. The presence of PFMS is associated with lower surgical success rates. Patients with pre-operatively identified PFMS should be counseled regarding a higher risk of treatment failure.


Estimating doses from Aboriginal bush foods post-remediation of a uranium mine.
This paper presents a calculator to facilitate assessments of ingestion doses from Aboriginal bush foods. The calculator combines information on traditional diet and land use with radionuclide concentration ratios and ingestion dose coefficients to estimate doses. The calculator was applied to the planned remediation of Ranger uranium mine to derive a set of scaling factors between radionuclide activity concentrations in environmental media and ingestion dose from bush foods. The scaling factors can be used to estimate doses from bush foods once the post-remediation radiological conditions of the mine and surrounding environment are known.


"Appraisal of state-of-the-art" The 2021 Distinguished Service Award of the Safety Pharmacology Society: Reflecting on the past to tackle challenges ahead.
This appraisal of state-of-the-art manuscript highlights and expands upon the thoughts conveyed in the lecture of Dr. Jean-Pierre Valentin, recipient of the 2021 Distinguished Service Award of the Safety Pharmacology Society, given on the 2nd December 2021. The article reflects on the strengths, weaknesses, opportunities, and threats that surrounded the evolution of safety and secondary pharmacology over the last 3 decades with a particular emphasis on pharmaceutical drug development delivery, scientific and technological innovation, complexities of regulatory framework and people leadership and development. The article further built on learnings from past experiences to tackle constantly emerging issues and evolving landscape whilst being cognizant of the challenges facing these disciplines in the broader drug development and societal context.


Single-trial neuromagnetic analysis reveals somatosensory dysfunction in chronic Minamata disease.
Methylmercury pollution is a global problem, and Minamata disease (MD) is a stark reminder that exposure to methylmercury can cause irreversible neurological damage. A "glove and stocking type" sensory disturbance due to injured primary sensory cortex (SI) (central somatosensory disturbance) is the most common neurologic sign in MD. As this sign is also prevalent in those with polyneuropathy, we aimed to develop an objective assessment for detecting central somatosensory disturbances in cases of chronic MD. We selected 289 healthy volunteers and 42 patients with MD. We recorded the sensory nerve action potentials (SNAPs) and somatosensory evoked magnetic fields (SEFs) to median nerve stimulation with magnetoencephalography. Single-trial epochs were classified into three categories (N20m, non-response, and P20m epochs) based on the cross-correlation between averaged sensor SEFs and individual epochs. We assessed SI responses (the appearance rate of P20m [P20m rate] and non-response epochs [non-response rate]) and early somatosensory cortical processing (N20m amplitude, reproducibility of N20m in single-trial responses [cross-correlation value], and induced gamma-band oscillations of the SI [gamma response] of single epochs excluding non-response epochs). Receiver operating characteristic curve analyses were used to examine the diagnostic accuracy of each parameter. We found that SNAPs exerted a marginal effect on the N20m. The N20m amplitude, cross-correlation value, and gamma response were significantly reduced in the MD group on either side (p < 0.0001), suggestive of altered early somatosensory cortical processing. Interestingly, the P20m rate and non-response rate were significantly increased in the MD group on either side (p < 0.0001), thereby suggesting impaired SI responses. Notably, P20m and absent N20m peaks were observed in 6 and 11 patients with MD, respectively, which may be attributed to increased numbers of P20m epochs. The cross-correlation value exhibited the highest correlation with the P20m rate or non-response rate. Thus, reduced reproducibility of N20m may play an important role in chronic MD. The cross-correlation value exhibited the highest correlation with the gamma response for both SI parameters in early somatosensory cortical processing. The area under the curve was > 0.77 (range: 0.77-0.79) for all parameters. Their confidence intervals overlapped with each other; thus, each SEF parameter likely had an approximately equivalent discrimination ability. In conclusion, chronic MD is characterized by impaired SI responses and alterations in early somatosensory cortical processing. Thus, single-trial neuromagnetic analysis of somatosensory function may be useful for detecting central somatosensory disturbance and elucidating the relevant pathophysiological mechanisms even in the context of chronic MD.


The effect of high-dose fentanyl anaesthesia on P50 in man.
The effect of high dosage fentanyl anaesthesia on P50 was studied in patients undergoing coronary bypass surgery. After induction of fentanyl anaesthesia 100 micrograms X kg-1 with pancuronium muscle relaxation P50 changed significantly (P less than 0.005) from 3.40 +/- 0.12 to 3.33 +/- 0.13 kPa. This anaesthesia technique decreases P50 in vivo, but this has more theoretical than practical importance.


The transformation of pediatric gliomatosis cerebri to cerebellar glioblastoma multiforme presenting as supra- and infratentorial acute disseminated encephalomyelitis. Case report.
Cerebellar glioblastoma multiforme (GBM) is a rare entity in adults and an extremely rare entity in children. Approximately 30 cases have been reported in the literature. The authors report the case of a histologically confirmed cerebellar GBM presenting initially as supra- and infratentorial gliomatosis cerebri. Acute disseminated encephalomyelitis had been diagnosed in the patient and that diagnosis remained until near the end of his treatment. This case underscores the need for recognizing the clinical presentation of gliomatosis cerebri and multifocal GBM in the pediatric subpopulation thought to harbor demyelinating disease.


Development of complexity categories for an investigational drug services complexity scoring tool to assess pharmacy effort in clinical trial initiation and maintenance.
Research pharmacy effort required to safely and compliantly manage investigational products (IP) varies between studies. No validated tool exists in the United States to evaluate these differences in effort. The Vizient Pharmacy Research Committee Investigational Drug Services (IDS) Subcommittee previously developed a systematic complexity scoring tool (CST) through expert consensus to assign a complexity score for pharmacy effort. This project seeks to develop and validate complexity categories based on CST scores. Vizient member institutions in IDS assigned a CST complexity score and a perceived complexity category (low, medium, or high) for study initiation and maintenance. Receiver operating characteristic (ROC) curve analysis defined the best CST score cutoff points for each complexity category. Comparing the CST-assigned to the user-perceived complexity category determined whether the CST-assigned complexity category aligned with practitioner assignment. A total of 322 responses were used to determine complexity score categories. The AUC values for study initiation and maintenance were 0.79 (P < 0.001) for the low/medium boundary and 0.80 (P < 0.001) for the medium/high boundary, suggesting the performance of the CST is good. The agreement between CST-assigned and user-perceived complexity categories was 60% for study initiation and 58% for maintenance. The Kendall rank correlation coefficient between the raters and ROC categories was strong, with a value of 0.48 for study initiation and 0.47 for maintenance. Development of the CST allows IDS pharmacies to objectively measure the complexity of clinical trials, which is a significant step towards assessing workload and guiding resource allocation.


A Human Randomized Controlled Trial Comparing Metabolic Responses to Single and Repeated Hypoglycemia in Type 1 Diabetes.
Hypoglycemia hinders optimal glycemic management in type 1 diabetes (T1D). Long diabetes duration and hypoglycemia impair hormonal counter-regulatory responses to hypoglycemia. Our study was designed to test whether (1) the metabolic responses and insulin sensitivity are impaired, and (2) whether they are affected by short-lived antecedent hypoglycemia in participants with T1D. In a randomized, crossover, 2x2 factorial design, 9 male participants with T1D and 9 comparable control participants underwent 30 minutes of hypoglycemia (p-glucose < 2.9 mmol/L) followed by a euglycemic clamp on 2 separate interventions: with and without 30 minutes of hypoglycemia the day before the study day. During both interventions insulin sensitivity was consistently lower, while counter-regulatory hormones were reduced, with 75% lower glucagon and 50% lower epinephrine during hypoglycemia in participants with T1D, who also displayed 40% lower lactate and 5- to 10-fold increased ketone body concentrations following hypoglycemia, whereas palmitate and glucose turnover, forearm glucose uptake, and substrate oxidation did not differ between the groups. In participants with T1D, adipose tissue phosphatase and tensin homolog (PTEN) content, hormone-sensitive lipase (HSL) phosphorylation, and muscle glucose transporter type 4 (GLUT4) content were decreased compared with controls. And antecedent hypoglycemic episodes lasting 30 minutes did not affect counter-regulation or insulin sensitivity. Participants with T1D displayed insulin resistance and impaired hormonal counter-regulation during hypoglycemia, whereas glucose and fatty acid fluxes were intact and ketogenic responses were amplified. We observed subtle alterations of intracellular signaling and no effect of short-lived antecedent hypoglycemia on subsequent counter-regulation. This plausibly reflects the presence of insulin resistance and implies that T1D is a condition with defective hormonal but preserved metabolic responsiveness to short-lived hypoglycemia.


Involvement of endothelin-1 in habitual exercise-induced increase in arterial compliance.
Habitual aerobic exercise results in a significant increase in central arterial compliance. Endothelin-1 (ET-1) is a potent endothelium-derived vasoconstrictor peptide and could play a role in mediating the habitual aerobic exercise-induced increase in central arterial compliance. The aim of the present study was to examine whether ET-1 is involved in the mechanisms underlying the increase in central arterial compliance with aerobic exercise training. Seven apparently healthy middle-aged and older (60 +/- 3 years) adults underwent systemic endothelin-A/B (ET(A/B))-receptor blockade (500 mg of Tracleer) before and after 12 weeks of aerobic exercise training (70 +/- 1% of maximal heart rate, 44 +/- 2 min day(-1), 4.4 +/- 0.1 days week(-1)). Basal carotid arterial compliance (via simultaneous B-mode ultrasound and arterial applanation tonometry on the common carotid artery) increased significantly after exercise training. Resting plasma ET-1 concentration decreased significantly after exercise training. Before exercise intervention, carotid arterial compliance increased significantly with the administration of the ET(A/B)-receptor blockade. After training, however, increases in carotid arterial compliance previously observed with the ET(A/B)-receptor blockade before training were abolished. Regular aerobic exercise training enhances central arterial compliance in middle-aged and older humans. The increase in arterial compliance was associated with the corresponding reduction in plasma ET-1 concentration as well as the elimination of ET-1-mediated vascular tone. These results suggest that reductions in ET-1 may be an important mechanism underlying the beneficial effect of exercise training on central artery compliance.


The relationship between trait mindfulness and affective symptoms: A meta-analysis of the Five Facet Mindfulness Questionnaire (FFMQ).
Trait mindfulness appears to be related to lower levels of negative affective symptoms, but it remains uncertain which facets of mindfulness are most important in this relationship. Accordingly, the present meta-analysis examined studies reporting correlations between affective symptoms and trait mindfulness as assessed by the Five Facet Mindfulness Questionnaire. A comprehensive search yielded 148 eligible studies, comprising 157 distinct samples and 44,075 participants. The weighted mean correlation for affective symptoms and overall trait mindfulness was r = -0.53. Among mindfulness facets, Nonjudge (r = -0.48) and Act with Awareness (r = -0.47) demonstrated the largest correlations, followed by Nonreact (r = -0.33) and Describe (r = -0.29). Observe was not significantly correlated with affective symptoms. No significant differences in the strength of correlations were found between anxiety, depression and posttraumatic stress disorder (PTSD) symptoms, though symptoms of generalized anxiety disorder exhibited a weaker negative relationship with the Describe facet compared to PTSD symptoms. Describe also showed a stronger relationship with affective symptoms in Eastern samples compared to Western samples, whereas Western samples had a stronger relationship with Nonjudge. These results provide insight into the nature of the association between trait mindfulness and negative affect.


Nonsurgical Resolution of a Bucket Handle Meniscal Tear: A Case Report.
A bucket handle meniscal tear (BHMT) often causes intermittent locking of the affected knee as the displaced fragment flips into the intercondylar notch. Surgical correction is usually required. Rare spontaneous healing of BHMTs has been described, primarily in young patients. This case report describes a 43-year-old man with magnetic resonance imaging-confirmed medial BHMT who received 3 treatments of platelet-rich plasma injections in and around the meniscus within 7 months after the diagnosis of the tear. The patient reported resolution of pain with walking 8 months after the injury. Magnetic resonance imaging 10 months after the injury and arthroscopy 47 months after the injury showed complete resolution of the meniscal tear. V.


DIGE enables the detection of a putative serum biomarker of fungal origin in a mouse model of invasive aspergillosis.
Invasive aspergillosis (IA) is a major threat for immunocompromised patients. Diagnostic difficulties often delay specific treatment initiation, which increases mortality. Finding new biomarkers to improve and speed accurate diagnosis is thus vital. To investigate the ability of proteomic methods for discovering new biomarkers of IA, we used a DIGE approach to perform a proteomic analysis on both bronchoalveolar lavages (BAL) and sera at different time-points of infection in a mouse model of invasive pulmonary aspergillosis. Progression of the infection was monitored using a bioluminescent strain of Aspergillus fumigatus. Sera proteins were enriched using the ProteoMiner kit (Biorad). This method allowed us to identify a fungal protein, the A. fumigatus major allergen Asp f 2, in sera of mice one day after the infection. However, this fungal protein was not detected three days after the infection. Importantly, in BAL, this work provides evidence of an in vivo complement evasion mechanism through the cleavage of C3b into three fragments during aspergillosis. Finally, our results underlining the inflammatory host response to IA in both lung and blood compartments at different times of infection may provide new insights into the pathophysiology of this disease.


Routine clinical cardiovascular magnetic resonance in paediatric and adult congenital heart disease: patients, protocols, questions asked and contributions made.
Cardiovascular magnetic resonance (CMR) of patients with congenital heart disease (CHD) has become routine clinical practice. However, existing CMR protocols focus predominantly on patients with ischemic heart disease, and information is limited on the types of patient with CHD who benefit from CMR investigation, and in what ways. Therefore the aim of this study was to answer the questions: What type of patients were studied by CMR in a centre specializing in paediatric and adult CHD management? What questions were asked, which protocols were used and were the questions successfully answered? To answer these questions, we conducted a cohort study of all 362 patients that received routine clinical CMR during 2007 at the Department of Paediatric Cardiology and Congenital Heart Disease at the Deutsches Herzzentrum München. Underlying diagnosis was in 33% Fallot's tetralogy, 17% aortic coarctation, 8% Ebstein's disease, 6% Marfan's disease, 4% single ventricle with Fontan-like circulation, and 32% others. Median age was 26 years (7 days - 75 years). Ventricular volumes were assessed in 67% of the patients; flow in 74%; unknown anatomy only in 9%; specific individual morphology of known anatomy in 83%; myocardial fibrosis in 8%; stress-induced myocardial perfusion defects in 1%. Only in 3% of the cases the question could not be fully answered. Contrary to common belief, routine CMR of patients with CHD was not requested to address global anatomical questions so much as to clarify specific questions of morphology and function of known anatomy. The CMR protocols used differed markedly from those widely used in patients with ischemic heart disease.


Robot-assisted inguinal sentinel lymph node biopsy in primary yolk sac tumor of the vulva.
Primary yolk sac tumor (YST) of the vulva is extremely rare and only introduced in case reports [1]. For those, routine inguinal lymph node dissection (LND) in absence of palpable inguinal lymph node is questionable. To avoid complications of inguinal LND, such as lymphedema, inguinal sentinel lymph node biopsy (SLNB) can be considered [2,3]. Since recent studies reported feasibility of robot-assisted inguinal lymphadenectomy [4], we showed the surgical procedures of robot-assisted inguinal SLNB. A 33-year-old woman with vulvar YST was referred. Two 2 cm sized masses were founded in right vulvar area, and lymph node enlargement was detected in only right external iliac area on imaging studies. Thus, we planned robot-assisted surgical treatment including inguinal SLNB using the fluorescent dye indocyanine green (ICG) during surgery for the vulvar YST. First, we performed intradermal and peri-tumoral injection of 1 ml solution including 1.25 mg of ICG at each 2, 5, 7, and 10 o'clock position for the two tumors. Fifteen minutes later, three-arm robotic surgical system was installed on right thigh. Dissecting the subcutaneous space under the Scarpa's fascia towards the inguinal ligament, one fluorescent inguinal sentinel lymph node was identified and excised, which showed no tumor on pathologic examination (Fig. 1). The procedure time was 70 minutes, and she underwent subsequent robot-assisted pelvic and para-aortic lymphadenectomy and wide local excision of the vulva. Robot-assisted inguinal SLNB is feasible. It may be an alternative to inguinal LND for avoiding relevant complications in patients with vulvar malignancy.


Huge anthropogenic microbial load during southwest monsoon season in coastal waters of Kakinada, Bay of Bengal.
To examine the influence of anthropogenic activities on the marine ecosystem near the coastal waters of the port city, Kakinada, a study was conducted to investigate the abundance of heterotrophic, indicator and pathogenic bacteria during the spring inter monsoon (SIM) and southwest monsoon (SWM) seasons. A drastic change in the marine bacteria due to the input of allochthonous bacteria during SWM was noticed. An order of magnitude higher abundance of indicators (Escherichia coli and Enterococcus faecalis) and bacterial pathogens (Proteus mirabilis and Pseudomonas aeruginosa) was observed during SWM. In contrast, Chlorophyll-a, heterotrophic bacterial abundance, Aeromonas hydrophila and Klebsiella pneumoniae were higher during SIM. A significant increase in some of the indicator and pathogenic bacterial abundance due to moderate rainfall suggests that the improper drainage system in the city could spread these bacteria, posing a considerable threat to both environment and human health.


Concentrations of ethanol in rebreathed air of rats: correlation with the discriminative stimulus effects of ethanol.
We modified the method of Pohorecky and Brick (14) for determination of ethanol (ETOH) concentrations in rebreathed air of rats. Rats were injected with different doses (1-2 g/kg) of ETOH and both arterial blood and rebreathed air samples were collected at various time intervals (15-120 min) after administration. We found a very good correlation (r = .96) between ETOH concentrations in arterial blood and in rebreathed air; the blood/breath conversion factor (+/- SEM) was 3241 +/- 55. In the second part of the study, rats were trained to discriminate between IP administered ETOH (1.2 g/kg) and the saline vehicle (12 ml/kg). Training occurred 15 min after administrations. Once trained to reliably differentiate between ETOH/saline training sessions, different doses (0.6, 0.9 and 1.2 g/kg) of ETOH were examined at various time intervals (1, 7.5, 15, 30, 60, 120 and 240 min) after administrations on certain test days. The results indicated a good correlation (r = .65) between the discriminative stimulus effects of ETOH and the concentrations measured in rebreathed air. The behavioral effects as well as the concentrations of ETOH in rebreathed air have a fast onset. The peak occurred 7.5 min after injection, and both the stimulus effects and concentrations of ETOH were time- and dose-dependent.


Design, synthesis and biological evaluation of novel plumbagin derivatives as potent antitumor agents with STAT3 inhibition.
Based on the structure of signal transducer and activator of transcription 3 (STAT3), a series of 1,4-naphthoquinones derived from plumbagin (PL) with STAT3 inhibition potential were designed, synthesized, and biologically evaluated in vitro against several human cancer cell lines (MDA-MB-231, HepG2 and A549 cells) and three normal cells. The structure-activity relationship (SAR) and molecular docking result showed that the presence of hydroxyl group at C-5 of PL might interact with STAT3 in the form of hydrogen bonds, which is conducive to the binding of this kind structures with STAT3. Among the target compounds, 7a displayed the most potent inhibition against cancer cells and weaker cytotoxicity on normal cells than PL. The western bolting analysis showed that 7a could suppress the phosphorylation of STAT3 as well as the downstream genes instead of affecting its upstream tyrosine kinases (Src and JAK2) levels and p-STAT1 expression. Furthermore, molecular docking indicated that 7a bound to STAT3 more tightly than PL, and it could significantly induce the apoptosis of cancer cells in vitro. All these results may provide reference for the discovery of effective STAT3 inhibitors.


Characterization of various cell lines from different ampullary cancer subtypes and cancer associated fibroblast-mediated responses.
Ampullary cancer is a relatively rare form of cancer and usually treated by pancreatoduodenectomy, followed by adjuvant therapy. The intestinal subtype is associated with markedly improved prognosis after resection. At present, only few cell lines are available for in vitro studies of ampullary cancer and they have not been collectively characterized. We characterize five ampullary cancer cell lines by subtype maker expression, epithelial-mesenchymal transition (EMT) features, growth and invasion, drug sensitivity and response to cancer-associated fibroblast conditioned medium (CAF-CM). On the basis of EMT features, subtype marker expression, growth, invasion and drug sensitivity three types of cell lines could be distinguished: mesenchymal-like, pancreatobiliary-like and intestinal-like. Heterogeneous effects from the cell lines in response to CAF-CM, such as different growth rates, induction of EMT markers as well as suppression of intestinal differentiation markers were observed. In addition, proteomic analysis showed a clear difference in intestinal-like cell line from other cell lines. Most of the available AMPAC cell lines seem to reflect a poorly differentiated pancreatobiliary or mesenchymal-like phenotype, which is consistent to their origin. We suggest that the most appropriate cell line model for intestinal-like AMPAC is the SNU869, while others seem to reflect aggressive AMPAC subtypes.


The value of intraoperative isometry measurements in anterior cruciate ligament reconstruction: an in vivo correlation between substitute tension and length change.
With the objective to evaluate an "isometry" measurement in an anterior cruciate ligament (ACL) substitute, in vivo measurements were taken on 10 patients undergoing reconstruction due to chronic ACL deficiency. Change in intraarticular length of a 1.2-mm test ligament, measured with an isometer, was correlated to the tension created in the same test ligament after fixation, measured with a piezoelectric load cell. The knee was passively moved through the 0-100 degrees range with the patient under general anaesthesia. Good statistical and visual correlations between length change and tension curves were found in individual knees. A correlation between total length change and maximum tension, for all knees grouped, was also found. Individual knees showed large variation in ability of the tissue to absorb load, resulting in a wide range of N/mm ratios between length and tension. Three knees with an isolated ACL injury all showed the least length change, implying a better restoration of kinematics in the absence of associated injuries. In nine of 10 knees the length change pattern could be used to identify the location of the femoral drill channel, as determined on an intraoperative lateral projection of the knee. Intraoperative isometry measurement can be used to predict the tension pattern in the reconstructed knee, but not the magnitude of tension. It will be useful to the surgeon in avoiding an anterior femoral ligament insertion site, which might threaten the integrity of the graft by tension rise in flexion.


Maternity leave policies for residents: an overview of issues and problems.
This article examines evidence of the need for maternity leave policies in residency programs and reviews studies of such policies. The author concludes that communication to residents of an established maternity leave policy facilitates equitable treatment of residents and minimizes disruptions for their colleagues and patients. The development of maternity leave policies is discussed in the context of existing and proposed federal legislation and the current pressures on program director and hospital administrators to improve other aspects residency training. Strategies for building flexibility in residency programs are presented.


Coming to grips with life upside down: how myosin fiber type and metabolic properties of sloth hindlimb muscles contribute to suspensory function.
Sloths exhibit almost obligatory suspensory locomotion and posture. These behaviors require both strength and fatigue resistance, although we previously found muscle fiber type characteristics in the forelimbs of sloths that belied these initial expectations. Based on locomotor roles of the forelimbs versus hindlimbs in propulsion and braking, respectively, sloth hindlimb musculature should be adapted for force production and energy savings by a near homogeneous expression of slow myosin heavy chain (MHC) fibers. This hypothesis was tested by determining MHC fiber type (%) distribution and energy metabolism in the hindlimbs of three-toed (B. variegatus, N = 5) and two-toed (C. hoffmanni, N = 3) sloths. A primary expression of the slow MHC-1 isoform was found in the hindlimbs of both species. Slow MHC fiber type (%) was significantly greater in the flexors of B. variegatus, whereas expression of fast MHC-2A fibers was significantly greater in the extensors of C. hoffmannni. MHC-1 fibers were largest in cross-sectional area (CSA) and comprised the greatest %CSA in each muscle sampled from both species. Enzyme assays showed elevated activity for anaerobic enzymes (CK and LDH) compared with low-to-moderate activity for aerobic enzymes (3-HAD and CS), and only CK activity was related to body size. These findings emphasize a joint stabilization role by the hindlimbs during suspension, especially in smaller three-toed sloths, and suggest that larger two-toed sloths could have muscles further modified for greater power output and/or prolonged arboreal maneuvering. Moreover, modifications to muscle metabolism rather than MHC expression may be more reflective of functional adaptation in sloth limbs.


Regional differences in intraneuronal localization of alpha-fetoprotein in developing mouse brain.
Estrogen receptor-containing regions of limbic, hypothalamic and amygdaloid areas of the developing mouse brain exhibited little or no intraneuronal immunofluorescence reactivity for AFP in comparison with the bright fluorescence of adjacent regions. These regional differences in the topographic distribution of intraneuronal AFP may represent intrinsic differences in the uptake, turnover or immunoreactivity of the internalized protein, perhaps related to estrogen metabolism by such neurons.


A Canine Leptospirosis Clinical Case Due to Leptospira interrogans (Serogroup Icterohaemorrhagiae) in a Dog Kennel in Castelvetrano (Western Sicily, South Italy).
Leptospirosis is a worldwide widespread zoonosis caused by Leptospira genus. We report an acute leptospirosis case in a puppy housed at a municipal kennel and the subsequent diagnostic investigations carried out on all dogs housed in the kennel. Laboratory investigation included mainly a microagglutination test, real-time PCR, and multi-locus sequence typing (MLST) for Leptospira genus. Other agents of infection were excluded. The puppy resulted positive for Leptospira interrogans Icterohaemorrhagiae both with serological and molecular assays. All of the other 66 dogs in the kennel underwent clinical and laboratory investigations twice, 15 days apart. No other dog showed leptospirosis clinical signs. At the first sampling, eight dogs (12%) showed antibodies against Leptospira interrogans serogroup Icterohaemorragiae serovar Copenhageni. Real-time PCR on urine samples of seropositive dogs detected Leptospira spp. DNA in one sample, then identified as Leptospira interrogans serogroup Icterohaemorragiae by MLST. Fifteen days after, four of the previous seropositive dogs still showed antibodies against Leptospira spp. All urine samples collected from seropositive dogs were negative at real-time PCR. The study allowed the early confirmation of a Leptospirosis case and the identification of at least one asymptomatic carrier of pathogenic Leptospira spp. The prompt activation of all appropriate management measures allowed limiting and extinguishing the infection.


Assessment of glomerular filtration rate and effective renal plasma flow in cystic fibrosis.
A study was conducted to examine renal function in 10 healthy control subjects and eight patients with cystic fibrosis in stable condition. Sequential bolus injections of 99mTc-DTPA and 125I-OIH were administered to assess glomerular filtration rate and effective renal plasma flow, respectively. Blood was subsequently collected for 3 hours, and urine for 24 hours. Renal clearances of both radioisotope markers were virtually identical in patients and controls. Inasmuch as neither glomerular filtration rate nor effective renal plasma flow was enhanced in patients with cystic fibrosis, increased clearance of drugs in these patients is unlikely to be the result of enhanced glomerular filtration or tubular secretion.


Molecular analyses reveal high species diversity of trematodes in a sub-Arctic lake.
To identify trematode diversity and life-cycles in the sub-Arctic Lake Takvatn, Norway, we characterised 120 trematode isolates from mollusc first intermediate hosts, metacercariae from second intermediate host fishes and invertebrates, and adults from fish and invertebrate definitive hosts, using molecular techniques. Phylogenies based on nuclear and/or mtDNA revealed high species richness (24 species or species-level genetic lineages) and uncovered trematode diversity (16 putative new species) from five families typical in lake ecosystems (Allocreadiidae, Diplostomidae, Plagiorchiidae, Schistosomatidae and Strigeidae). Sampling potential invertebrate hosts allowed matching of sequence data for different stages, thus achieving molecular elucidation of trematode life-cycles and exploration of host-parasite interactions. Phylogenetic analyses also helped identify three major mollusc intermediate hosts (Radix balthica, Pisidium casertanum and Sphaerium sp.) in the lake. Our findings increase the known trematode diversity at the sub-Arctic Lake Takvatn, showing that digenean diversity is high in this otherwise depauperate sub-Arctic freshwater ecosystem and indicating that sub-Arctic and Arctic ecosystems may be characterised by unique trematode assemblages.


The distribution of two hnRNP-associated proteins defined by a monoclonal antibody is altered in heat-shocked HeLa cells.
A monoclonal antibody obtained after mice were immunized with hnRNP purified from HeLa cells recognizes two polypeptides of Mr 35,000 and 37,000. By immunocytofluorescence, these antigens can be visualized only in cells previously heat shocked at 45 degrees C for 5 or 10 min, although they are present at the same level in unstressed and stressed cells. The signal, which is mostly concentrated in the interchromatin space, where hnRNP fibrils are located, does not accumulate with time and disappears 4 to 5 h after heat shock. Discrimination between the two types of hnRNP substructures, the 30-50 S monoparticles and the nuclear matrix fibrils, based on differential sensitivity to salt or ribonuclease treatment, showed that in unstressed cells the antigens behave as monoparticle proteins. In contrast, in heat-shocked cells, most 35-37K antigens behave as nuclear matrix proteins. Thus, heat shock seems to induce a rapid and reversible switch of these two antigens from hnRNP monoparticles to the nuclear matrix. The data demonstrate that heat shock, which was previously shown not to alter the overall RNA: protein packaging ratio of hnRNP, induces subtle modifications of their substructure. Such modifications might be of importance since heat shock is known for instance to affect pre-mRNA processing.


Effects of anticipated emotional category and temporal predictability on the startle reflex.
Anticipated emotional category and temporal predictability are key characteristics that have both been shown to impact psychophysiological indices of defensive motivation (e.g., the startle reflex). To date, research has primarily examined these features in isolation, and it is unclear whether they have additive or interactive effects on defensive motivation. In the present study, the startle reflex was measured in anticipation of low arousal neutral, moderate arousal pleasant, and high arousal unpleasant pictures that were presented with either predictable or unpredictable timing. Linear mixed-effects modeling was conducted to examine startle magnitude across time, and the intercept at the beginning and end of the task. Across the entire task, the anticipation of temporally unpredictable (relative to predictable) pictures and emotional (relative to neutral) pictures potentiated startle magnitude, but there was no interaction between the two features. However, examination of the intercept at the beginning of the task indicated a Predictability by Emotional Category interaction, such that temporal unpredictability enhanced startle potentiation in anticipation of unpleasant pictures only. Examination of the intercept at the end of the task indicated that the effects of predictability and emotional category on startle magnitude were largely diminished. The present study replicates previous reports demonstrating that emotional category and temporal predictability impact the startle reflex, and provides novel evidence suggesting an interactive effect on defensive motivation at the beginning of the task. This study also highlights the importance of examining the time course of the startle reflex.


Headspace solid phase microextraction/gas chromatography-mass spectrometry combined to chemometric analysis for volatile organic compounds determination in canine hair: a new tool to detect dog contamination by visceral leishmaniasis.
A new analytical methodology using HS-SPME/GC-MS was optimized in order to attain maximum sensitivity, using multivariate strategies. The proposed method was employed to evaluate the VOC profile exhaled from canine hair samples collected from 8 healthy dogs and from 16 dogs infected by Leishmania infantum. 274 VOCs were detected, which could be identified as aldehydes, ketones and hydrocarbons. After application of the Soft Independent Modeling of Class Analogy (SIMCA) and Principal Component Analysis (PCA) healthy and infected dogs, with similar VOCs profiles, could be separately grouped, based on compounds such as 2-hexanone, benzaldehyde, and 2,4-nonadienal. The proposed method is non-invasive, painless, readily accepted by dog owners and could be useful to identify several biomarkers with applications in the diagnosis of diseases.


18F-fluorodeoxyglucose positron emission tomography in dementia with Lewy bodies.
Among individuals with dementia with Lewy bodies, pathologic correlates of clinical course include the presence and extent of coexisting Alzheimer's pathology and the presence of transitional or diffuse Lewy body disease. The objectives of this study are to determine (i) whether 18F-fluorodeoxyglucose PET signature patterns of dementia with Lewy bodies are associated with the extent of coexisting Alzheimer's pathology and the presence of transitional or diffuse Lewy body disease and (ii) whether these 18F-fluorodeoxyglucose pattern(s) are associated with clinical course in dementia with Lewy bodies. Two groups of participants were included: a pathology-confirmed subset with Lewy body disease (n = 34) and a clinically diagnosed group of dementia with Lewy bodies (n = 87). A subset of the clinically diagnosed group was followed longitudinally (n = 51). We evaluated whether 18F-fluorodeoxyglucose PET features of dementia with Lewy bodies (higher cingulate island sign ratio and greater occipital hypometabolism) varied by Lewy body disease subtype (transitional versus diffuse) and Braak neurofibrillary tangle stage. We investigated whether the PET features were associated with the clinical trajectories by performing regression models predicting Clinical Dementia Rating Scale Sum of Boxes. Among autopsied participants, there was no difference in cingulate island sign or occipital hypometabolism by Lewy body disease type, but those with a lower Braak tangle stage had a higher cingulate island sign ratio compared to those with a higher Braak tangle stage. Among the clinically diagnosed dementia with Lewy bodies participants, a higher cingulate island ratio was associated with better cognitive scores at baseline and longitudinally. A higher 18F-fluorodeoxyglucose PET cingulate island sign ratio was associated with lower Braak tangle stage at autopsy, predicted a better clinical trajectory in dementia with Lewy body patients and may allow for improved prognostication of the clinical course in this disease.


Mechanical performance of traditional distraction-based dual growing rod constructs.
Growing rod constructs are an important contribution in the treatment of children with early onset scoliosis even though these devices experience high rates of rod fracture. The mechanical performance of traditional, distraction-based dual growing rod constructs is not well understood, and mechanical models for predicting device performance are limited. Two mechanical models were developed and used to determine the mechanical performance of various growing rod configurations by increasing construct complexity. Mechanical bench testing and finite element (FE) analysis. Static and dynamic compression bending tests were based on an ASTM F1717 method modified to accommodate dual growing rod constructs. Six construct configurations were tested, mechanical properties were recorded, and statistical analyses were performed to determine significant differences between groups: (1) no connectors (rods only), (2) side-by-side connectors, (3) side-by-side connectors plus 4 crosslinks, (4) (40-mm long tandem connectors, (5) 80-mm long tandem connectors, and (6) 80-mm long tandem connectors plus 4 crosslinks. FE analysis was used to predict the stress distribution within the constructs. The static results indicated greater stiffness, yield load, and peak load as the axial connector length increased (side-by-side to 40 mm tandem to 80 mm tandem). The dynamic results showed similar cycles to failure for side-by-side and tandem connector (40 and 80 mm) construct configurations without crosslinks. Crosslinks shifted the location of rod fracture observed experimentally and significantly reduced the fatigue life of the construct. The flexibility of the construct decreased significantly as the axial connector length increased. FE predictions were highly consistent with the experimentally measured values and provided information on stress distribution within the rod for comparison to experimental fracture locations. This is the first study to evaluate mechanical performance of various configurations of pediatric growing rod constructs using preclinical models. The current study is consistent with a previous retrieval study in that rigid constructs lacking flexibility (ie, higher stiffness and lower displacement), such as those with 80-mm tandem connectors and multiple crosslinks, demonstrated decreased mechanical performance as shown through both experimental and computational models. Additionally, the experimental and computational findings suggest that surgeons should strategically consider the number of interconnecting components and subsequent stress concentrations along the posterior side of the rod. For example, changing the placement of crosslinks to low stress regions of the construct or not using crosslinks in the construct are options.


Characteristics of patients with lung cancer under the age of 45 years: a case control study.
The aim of this study was to define the clinical and pathological features associated with lung cancer diagnosed in persons under the age of 45 compared with lung cancer in an older population of greater than 45 years. A case control study was undertaken. Cases were defined as patients diagnosed with lung cancer under the age of 45 years. Controls were lung cancer patients over 45 years matched only for the date of diagnosis. Up to four controls were selected for each case. A retrospective review was undertaken of the records of a single tertiary respiratory institution which served a population of 1.2 million. Forty-eight cases of lung cancer (< 45 years of age) were identified and were compared to 123 matched controls with lung cancer (> 45 years of age). Of the cases 67% were female compared with 32% female cases in the control group (P < 0.01). The rate of adenocarcinoma was significantly higher among cases (48% vs 27%, P=0.001), while squamous cell carcinoma was more common in the controls (35% vs 17% P =0.001). Smoking was common in both groups although less so among cases (79% vs 95%, P=0.001)). There was no difference in survival rates between the cases (11 months) compared with controls (8 months) (P=0.1). Young lung cancer patients were predominantly female and adenocarcinoma accounted for a disproportionate number of the histological types. Cigarette smoking appears to be the main aetiological agent but as 21% of the patients under 45 years were non-smokers, other factors (genetic/hormonal) may also contribute. This study has not demonstrated a worse outcome in younger patients.


Parents' reactions at 24-48 hrs after a preschool child's head injury.
Our objectives were, first, to compare mothers' and fathers' early reactions (stressors, concerns) to the preschool child's head injury, their perceptions of the child's injury severity, and their social support and mental health; second, to compare families with a child in the pediatric intensive care unit (PICU) vs. general care unit (GCU) on these variables; and third, to describe the relationships between parents' early reactions and perceived and objective injury severity, their social support, and mental health. Analysis of data collected in the hospital 24-48 hrs after the child's admission as part of a longitudinal study of parent and family functioning after a preschool child's head injury. Seven tertiary care centers: three free-standing children's hospitals and four comprehensive hospitals. Participants were 182 mothers and 64 fathers of 183 preschool children (ages 3-6) hospitalized for head injury, half in a PICU. Data collection. We measured parents' early reactions (stressors, concerns), influenced by parent mental health, social support, and objective and perceived injury severity. Mothers reported more stress than fathers regarding the child's behavior and emotions, communication with staff, and their parental role. Mothers in the PICU group reported more concern about the child's future and more stress regarding the child's appearance, sights and sounds of the unit, and procedures done to the child than mothers in the GCU group. Fathers in the PICU and GCU groups reported similar levels of stress and concern. Mothers' reactions were influenced by objective and perceived injury severity, social support, and psychological distress. Fathers' reactions were influenced by objective injury severity and psychological distress. Although mother-father couples rated their child's injury severity similarly, mothers experienced more stress than fathers. Social support decreased the stress for mothers but not for fathers. The experience of pediatric head trauma was more stressful for mothers of children in the PICU than mothers of children in the GCU.


Radiosensitizing effect of a phenylbutyrate-derived histone deacetylase inhibitor in hepatocellular carcinoma.
Radiotherapy is integrated into the multimodal treatment of localized hepatocellular carcinoma (HCC) refractory to conventional treatment. Tumor control remains unsatisfactory and the sublethal effect associates with secondary spread. The use of an effective molecularly targeted agent in combination with radiotherapy is a potential therapeutic approach. Our aim was to assess the effect of combining a phenylbutyrate-derived histone deacetylase (HDAC) inhibitor, AR-42, with radiotherapy in in vitro and in vivo models of human HCC. Human HCC cell lines (Huh-7 and PLC-5) were used to evaluate the in vitro synergism of combining AR-42 with irradiation. Flow cytometry analyzed the cell cycle changes, whereas Western blot investigated the protein expressions after the combined treatment. Severe combined immunodeficient (SCID) mice bearing ectopic and orthotopic HCC xenografts were treated with AR-42 and/or radiotherapy for the in vivo response. AR-42 significantly enhanced radiation-induced cell death by the inhibition of the DNA end-binding activity of Ku70, a highly versatile regulatory protein for DNA repair, telomere maintenance, and apoptosis. In ectopic xenografts of Huh-7 and PLC-5, pretreatment with AR-42 significantly enhanced the tumor-suppressive effect of radiotherapy by 48% and 66%, respectively. A similar combinatorial effect of AR-42 (10 and 25 mg/kg) and radiotherapy was observed in Huh-7 orthotopic model of tumor growth by 52% and 82%, respectively. This tumor suppression was associated with inhibition of intratumoral Ku70 activity as well as reductions in markers of HDAC activity and proliferation, and increased apoptosis. AR-42 is a potent, orally bioavailable inhibitor of HDAC with therapeutic value as a radiosensitizer of HCC.


Brain structure of perinatally HIV-infected patients on long-term treatment: A systematic review.
We aim to give an overview of the available evidence on brain structure and function in PHIV-infected patients (PHIV+) using long-term combination antiretroviral therapy (cART) and how differences change over time. We conducted an electronic search using MEDLINE, Embase, and PsycINFO. We used the following selection criteria: cohort and cross-sectional studies that reported on brain imaging differences between PHIV+ of all ages who used cART for at least six months before neuroimaging and HIV-negative controls. Two reviewers independently selected studies, performed data extraction, and assessed quality of studies. After screening 1500 abstracts and 343 full-text articles, we identified 19 eligible articles. All included studies had a cross-sectional design and used MRI with different modalities: structural MRI (n = 7), diffusion tensor imaging (DTI) (n = 6), magnetic resonance spectroscopy (n = 5), arterial spin labeling (n = 1), and resting-state functional neuroimaging (n = 1). Studies showed considerable methodological limitations and heterogeneity, preventing us to perform meta-analyses. DTI data on white matter microstructure suggested poorer directional diffusion in cART-treated PHIV+ compared with controls. Other modalities were inconclusive. Evidence may suggest brain structure and function differences in the population of PHIV+ on long-term cART compared with the HIV-negative population. Because of a small study population, and considerable heterogeneity and methodological limitations, the extent of brain structure and function differences on neuroimaging between groups remains unknown.


Degenerative changes of vein grafts in preparation media: preliminary studies by electron microscopy and fibrinolytic autography.
Degenerative changes of saphenous vein grafts in four preparation media (heparinized whole blood at room temperature and 4 degrees C, and heparinized normal saline at same temperatures) were examined by scanning and transmission electron microscopy and fibrinolytic autography. Following 60--90 min. storage in heparinized normal saline at room temperature, marked morphological changes were present in the media, accompanied by swelling of the endothelial cells, however the tunica media and adventitia were well preserved even after 120 minutes in all of the four preparation media. The decrease in fibrinolytic activity was comparable to the observed morphological changes. In heparinized whole blood at 4 degrees C, degenerative changes were slow and mostly of a slight nature.