Histopathological and immunohistochemical investigation of the effect of Shilajit in rats with experimental spinal cord injury.
This experimental study was designed to investigate the histopathological and immunohistochemical effects of Shilajit in rats with experimentally induced spinal cord injury (SCI). The rats were divided into three groups: Control group: The group in which spinal cord damage was created but no drug was administered. Low-dose group: This is the group in which intraperitoneal Shilajit is given at a dose of 150 mg/kg at the 1st h, 1st day, 2nd day, and 3rd day after spinal cord damage was induced. High-dose group: This is the group in which intraperitoneal Shilajit is given at a dose of 250 mg/kg at the 1st h, 1st day, 2nd day, and 3rd day after spinal cord damage was induced. Thin sections taken from the spinal cord after euthanasia were sent for histopathological and immunohistochemical examination. Histopathological examination of the high-dose group showed lower amounts of morphological findings compared to the low-dose group and control group. While a significant CD68 immune reaction was observed in the control group of rats with spinal injury, the positive immune reaction was found to be significantly decreased in the Shilajit-applied groups. It is thought that the use of Shilajit in SCI will reduce the effects of secondary damage in SCI and that its administra-tion to such patients will have positive effects on the results.


Prostaglandin E2 attenuates preoptic expression of GABAA receptors via EP3 receptors.
Prostaglandin E(2) (PGE(2)) has been shown to produce fever by acting on EP3 receptors within the preoptic area of the brain. However, there is little information about the molecular events downstream of EP3 activation in preoptic neurons. As a first step toward this issue, we examined PGE(2)-induced gene expression changes at single-cell resolution in preoptic neurons expressing EP3. Brain sections of the preoptic area from PGE(2)- or saline-injected rats were stained with an anti-EP3 antibody, and the cell bodies of EP3-positive neurons were dissected and subjected to RNA amplification procedures. Microarray analysis of the amplified products demonstrated the possibility that gene expression of gamma-aminobutyric acid type A (GABA(A)) receptor subunits is decreased upon PGE(2) injection. Indeed, we found that most EP3-positive neurons in the mouse preoptic area are positive for the alpha2 or gamma2 GABA(A) receptor subunit. Moreover, PGE(2) decreased the preoptic gene expression of these GABA(A) subunits via an EP3-dependent and pertussis toxin-sensitive pathway. PGE(2) also attenuated the preoptic protein expression of the alpha2 subunit in wild-type but not in EP3-deficient mice. These results indicate that PGE(2)-EP3 signaling elicits G(i/o) activation in preoptic thermocenter neurons, and we propose the possibility that a rapid decrease in preoptic GABA(A) expression may be involved in PGE(2)-induced fever.


Multiple juvenile polyposis. A study of the pathogenesis of juvenile polyps and their relationship to colonic adenomas.
Solitary juvenile polyps are common lesions whose pathogenesis is poorly understood. Multiple juvenile polyposis is characterized by large numbers of these lesions either confined to the colon or throughout the gastrointestinal tract. A study of two cases of multiple juvenile polyposis provided fresh insight into the pathogenesis of juvenile polyps and their relationship to colonic adenomas. Mucosal ulceration in very early lesions, together with glandular epithelial calcification, suggested that impaired cell renewal resulting from disturbed regenerative kinetics may predispose to surface epithelial erosion, setting in motion a cycle of ulceration, inflammation, and granulation tissue formation. We postulate that a dyskinetic continuum may link juvenile, "metaplastic," and adenomatous polyps. The finding in our second case of multiple adenomatous lesions, including a villoglandular polyp, emphasizes the neoplastic potential of juvenile polyposis.


Hydrogen evolution in [NiFe] hydrogenases and related biomimetic systems: similarities and differences.
In this work, a detailed quantum chemical study of the mechanism of [Ni(bdt)(dppf)] (Ni(II)L) catalyzed hydrogen formation [A. Gan, T. L. Groy, P. Tarakeshwar, S. K. S. Mazinani, J. Shearer, V. Mujica and A. K. Jones, J. Am. Chem. Soc., 2015, 137, 1109-1115] following an electro-chemical-electro-chemical (ECEC) pathway is reported. The complex exclusively catalyzes the reduction of protons to molecular hydrogen. The calculations suggest that the first one-electron reduction of the [Ni(II)L] catalyst is the rate limiting step of the catalytic cycle and hence, the buildup of detectable reaction intermediates is not expected. The catalytic activity of the [Ni(II)L] complex is facilitated by the flexibility of the ligand system, which allows the ligand framework to adapt to changes in the Ni oxidation state over the course of the reaction. Additionally, a comparison is made with the catalytic activity of [NiFe] hydrogenase. It is argued that the directionality of the reversible hydrogen formation reaction is controlled by the ligand field of the nickel ion and the possibility for side-on (η(2)) binding of H2: if the ligand framework does not allow for η(2) binding of H2, as is the case for [Ni(II)L], the catalyst irreversibly reduces protons. If the ligand field allows η(2) binding of H2, the catalyst can in principle work reversibly. The conditions for η(2) binding are discussed.


Quantitative 1H nuclear magnetic resonance metabolite profiling as a functional genomics platform to investigate alkaloid biosynthesis in opium poppy.
Opium poppy (Papaver somniferum) produces a diverse array of bioactive benzylisoquinoline alkaloids and has emerged as a versatile model system to study plant alkaloid metabolism. The plant is widely cultivated as the only commercial source of the narcotic analgesics morphine and codeine. Variations in plant secondary metabolism as a result of genetic diversity are often associated with perturbations in other metabolic pathways. As part of a functional genomics platform, we used (1)H nuclear magnetic resonance (NMR) metabolite profiling for the analysis of primary and secondary metabolism in opium poppy. Aqueous and chloroform extracts of six different opium poppy cultivars were subjected to chemometric analysis. Principle component analysis of the (1)H NMR spectra for latex extracts clearly distinguished two varieties, including a low-alkaloid variety and a high-thebaine, low-morphine cultivar. Distinction was also made between pharmaceutical-grade opium poppy cultivars and a condiment variety. Such phenotypic differences were not observed in root extracts. Loading plots confirmed that morphinan alkaloids contributed predominantly to the variance in latex extracts. Quantification of 34 root and 21 latex metabolites, performed using Chenomx NMR Suite version 4.6, showed major differences in the accumulation of specific alkaloids in the latex of the low-alkaloid and high-thebaine, low-morphine varieties. Relatively few differences were found in the levels of other metabolites, indicating that the variation was specific for alkaloid metabolism. Exceptions in the low-alkaloid cultivar included an increased accumulation of the alkaloid precursor tyramine and reduced levels of sucrose, some amino acids, and malate. Real-time polymerase chain reaction analysis of 42 genes involved in primary and secondary metabolism showed differential gene expression mainly associated with alkaloid biosynthesis. Reduced alkaloid levels in the condiment variety were associated with the reduced abundance of transcripts encoding several alkaloid biosynthetic enzymes.


Antimicrobial activity of the essential oil from Lippia sidoides, carvacrol and thymol against oral pathogens.
Dental caries and periodontal disease are associated with oral pathogens. Several plant derivatives have been evaluated with respect to their antimicrobial effects against such pathogenic microorganisms. Lippia sidoides Cham (Verbenaceae), popularly known as "Alecrim-pimenta" is a typical shrub commonly found in the Northeast of Brazil. Many plant species belonging to the genus Lippia yield very fragrant essential oils of potential economic value which are used by the industry for the commercial production of perfumes, creams, lotions, and deodorants. Since the leaves of L. sidoides are also extensively used in popular medicine for the treatment of skin wounds and cuts, the objective of the present study was to evaluate the composition and antimicrobial activity of L. sidoides essential oil. The essential oil was obtained by hydro-distillation and analyzed by GC-MS. Twelve compounds were characterized, having as major constituents thymol (56.7%) and carvacrol (16.7%). The antimicrobial activity of the oil and the major components was tested against cariogenic bacterial species of the genus Streptococcus as well as Candida albicans using the broth dilution and disk diffusion assays. The essential oil and its major components thymol and carvacrol exhibited potent antimicrobial activity against the organisms tested with minimum inhibitory concentrations ranging from 0.625 to 10.0 mg/mL. The most sensitive microorganisms were C. albicans and Streptococcus mutans. The essential oil of L. sidoides and its major components exert promising antimicrobial effects against oral pathogens and suggest its likely usefulness to combat oral microbial growth.


Magnetic-field-induced soft-mode quantum phase transition in the high-temperature superconductor La1.855Sr0.145CuO4: an inelastic neutron-scattering study.
Inelastic neutron-scattering experiments on the high-temperature superconductor La1.855Sr0.145CuO4 reveal a magnetic excitation gap Delta that decreases continuously upon application of a magnetic field perpendicular to the CuO2 planes. The gap vanishes at the critical field required to induce long-range incommensurate antiferromagnetic order, providing compelling evidence for a field-induced soft-mode driven quantum phase transition.


Identification of a functionally important negatively charged residue within the second catalytic site of the SUR1 nucleotide-binding domains.
The ATP-sensitive K+ channel (KATP channel) couples glucose metabolism to insulin secretion in pancreatic beta-cells. It is comprised of sulfonylurea receptor (SUR)-1 and Kir6.2 proteins. Binding of Mg nucleotides to the nucleotide-binding domains (NBDs) of SUR1 stimulates channel opening and leads to membrane hyperpolarization and inhibition of insulin secretion. To elucidate the structural basis of this regulation, we constructed a molecular model of the NBDs of SUR1, based on the crystal structures of mammalian proteins that belong to the same family of ATP-binding cassette transporter proteins. This model is a dimer in which there are two nucleotide-binding sites, each of which contains residues from NBD1 as well as from NBD2. It makes the novel prediction that residue D860 in NBD1 helps coordinate Mg nucleotides at site 2. We tested this prediction experimentally and found that, unlike wild-type channels, channels containing the SUR1-D860A mutation were not activated by MgADP in either the presence or absence of MgATP. Our model should be useful for designing experiments aimed at elucidating the relationship between the structure and function of the KATP channel.


Genetic and Environmental Influences on Eating Behavior - A Study of Twin Pairs Reared Apart or Reared Together.
This study examined the relative influence of genetic versus environmental factors on specific aspects of eating behavior. Adult monozygotic twins (22 pairs and 3 singleton reared apart, 38 pairs and 9 singleton reared together, age 18-76 years, BMI 17-43 kg/m2) completed the Three Factor Eating Questionnaire. Genetic and environmental variance components were determined for the three eating behavior constructs and their subscales using model-fitting univariate and multivariate analyses. Unique environmental factors had a substantial influence on all eating behavior variables (explaining 45-71% of variance), and most strongly influenced external locus for hunger and strategic dieting behavior of restraint (explaining 71% and 69% of variance, respectively). Genetic factors had a statistically significant influence on only 4 variables: restraint, emotional susceptibility to disinhibition, situational susceptibility to disinhibition, and internal locus for hunger (heritabilities were 52%, 55%, 38% and 50%, respectively). Common environmental factors did not statistically significantly influence any variable assessed in this study. In addition, multivariate analyses showed that disinhibition and hunger share a common influence, while restraint appears to be a distinct construct. These findings suggest that the majority of variation in eating behavior variables is associated with unique environmental factors, and highlights the importance of the environment in facilitating specific eating behaviors that may promote excess weight gain.


Needle-based confocal laser endomicroscopy for real-time diagnosing and staging of lung cancer.
Diagnosing lung cancer in the absence of endobronchial abnormalities is challenging. Needle-based confocal laser endomicroscopy (nCLE) enables real-time microscopic imaging of cells. We assessed the feasibility and safety of using nCLE for real-time identification of lung cancer.In patients with suspected or proven lung cancer scheduled for endoscopic ultrasound (EUS), lung tumours and mediastinal lymph nodes were imaged with nCLE before fine-needle aspiration (FNA) was performed. nCLE lung cancer characteristics were identified by comparison with pathology. Multiple blinded raters validated CLE videos of lung tumours and mediastinal nodes twice.EUS-nCLE-FNA was performed in 22 patients with suspected or proven lung cancer in whom 27 lesions (six tumours, 21 mediastinal nodes) were evaluated without complications. Three nCLE lung cancer criteria (dark enlarged pleomorphic cells, dark clumps and directional streaming) were identified. The accuracy of nCLE imaging for detecting malignancy was 90% in tumours and 89% in metastatic lymph nodes. Both inter-observer agreement (mean κ=0.68, 95% CI 0.66-0.70) and intra-observer agreement (mean±sd κ=0.70±0.15) were substantial.Real-time lung cancer detection by endosonography-guided nCLE was feasible and safe. Lung cancer characteristics were accurately recognised.


Diagnostic concordance between mobile interfaces and conventional workstations for emergency imaging assessment.
Mobile devices and software are now available with sufficient computing power, speed and complexity to allow for real-time interpretation of radiology exams. In this paper, we perform a multivariable user study that investigates concordance of image-based diagnoses provided using mobile devices on the one hand and conventional workstations on the other hand. We performed a between-subjects task-analysis using CT, MRI and radiography datasets. Moreover, we investigated the adequacy of the screen size, image quality, usability and the availability of the tools necessary for the analysis. Radiologists, members of several teams, participated in the experiment under real work conditions. A total of 64 studies with 93 main diagnoses were analyzed. Our results showed that 56 cases were classified with complete concordance (87.69%), 5 cases with almost complete concordance (7.69%) and 1 case (1.56%) with partial concordance. Only 2 studies presented discordance between the reports (3.07%). The main reason to explain the cause of those disagreements was the lack of multiplanar reconstruction tool in the mobile viewer. Screen size and image quality had no direct impact on the mobile diagnosis process. We concluded that for images from emergency modalities, a mobile interface provides accurate interpretation and swift response, which could benefit patients' healthcare.


Endoscopic endonasal transsphenoidal approach to large and giant pituitary adenomas: institutional experience and predictors of extent of resection.
OBJECT.: While the use of endoscopic approaches has become increasingly accepted in the resection of pituitary adenomas, limited evidence exists regarding the success of this technique for patients with large and giant pituitary adenomas. This study reviews the outcomes of a large cohort of patients with large and giant pituitary adenomas who underwent endoscopic endonasal transsphenoidal surgery at the authors' institution and focuses on identifying factors that can predict extent of resection and hence aid in developing guidelines and indications for the use of endoscopic endonasal transsphenoidal surgery versus open craniotomy approaches to large and giant pituitary adenomas. The authors reviewed 487 patients who underwent endoscopic endonasal transsphenoidal resection of sellar masses. From this group, 73 consecutive patients with large and giant pituitary adenomas (defined as maximum diameter ≥ 3 cm and tumor volume ≥ 10 cm(3)) who underwent endoscopic endonasal transsphenoidal surgery between January 1, 2006, and June 6, 2012, were included in the study. Clinical presentation, radiological studies, laboratory investigations, tumor pathology data, clinical outcomes, extent of resection measured by volumetric analysis, and complications were analyzed. The mean preoperative tumor diameter in this series was 4.1 cm and the volume was 18 cm(3). The average resection rate was 82.9%, corresponding with a mean residual volume of 3 cm(3). Gross-total resection was achieved in 16 patients (24%), near-total in 11 (17%), subtotal in 24 (36%), and partial in 15 (23%). Seventy-three percent of patients experienced improvement in visual acuity, while 24% were unchanged. Visual fields were improved in 61.8% and unchanged in 5.5%. Overall, 27 patients (37%) experienced a total of 32 complications. The most common complications were sinusitis (14%) and CSF leak (10%). Six patients underwent subsequent radiation therapy because of aggressive tumor histopathology. No deaths occurred in this cohort of patients. Statistically significant predictors of extent of resection included highest Knosp grade (p = 0.001), preoperative tumor volume (p = 0.025), preoperative maximum tumor diameter (p = 0.002), hemorrhagic component (p = 0.027), posterior extension (p = 0.001), and sphenoid sinus invasion (p = 0.005). Endoscopic endonasal transsphenoidal surgery is an effective treatment method for patients with large and giant pituitary adenomas, which results in high (> 80%) rates of resection and improvement in visual function. It is not associated with high rates of major complications and is safe when performed by experienced surgeons. The preoperative Knosp grade, tumor volume, tumor diameter, hemorrhagic components on MRI, posterior extension, and sphenoid sinus invasion may allow a prediction of extent of resection and in these patients a staged operation may be required to maximize extent of resection.


Pupillary responses and airway reactivity in asthma.
Pupillary responses to phenylephrine and carbachol and airway reactivity were determined in 19 drug-free persons with mild allergic asthma, 21 nonatopic control subjects, and six asymptomatic atopic control subjects. The concentration of phenylephrine required to dilate the pupil 1 mm, obtained by interpolation on the dose-response curve, was taken as a measure of alpha-adrenergic sensitivity. This concentration was significantly lower for subjects with asthma than for atopic or nonatopic control subjects (1.11 +/- 0.68% [mean +/- SD], 1.86 +/- 0.81%, and 1.52 +/- 0.67%, respectively; p less than 0.05 by analysis of variance). The concentration of carbachol required for pupillary constriction of 1 mm, obtained by interpolation on the dose-response curve, was taken as a measure of cholinergic sensitivity. This value was significantly lower for subjects with asthma than for atopic or nonatopic control subjects (0.23 +/- 0.12%, 0.41 +/- 0.14%, and 0.35 +/- 0.19%, respectively; p less than 0.05 by analysis of variance). Pupillary alpha-adrenergic and cholinergic sensitivity were significantly correlated (Spearman's rank correlation coefficient, Rs = 0.42, p less than 0.05). Both alpha-adrenergic and cholinergic sensitivity of the pupils correlated with airway reactivity expressed as concentration of methacholine causing a 20% fall in FEV1 (Rs = 0.35, p less than 0.05, and Rs = 0.45, p less than 0.05, respectively). The autonomic aberrations cannot be attributed to drug use or significantly abnormal pulmonary mechanics. Autonomic abnormalities are associated with allergic asthma and may contribute to the airway reactivity characteristic of the disease.


Therapeutic effect of an ayahuasca analogue in clinically depressed patients: a longitudinal observational study.
Studies have suggested mental health improvements following the use of the psychotropic plant concoction ayahuasca in non-clinical and clinical samples. The present observational study assessed depressive symptomatology in 20 clinically depressed patients (symptom score > 13 on the Beck's Depression Inventory) before attendance of an ayahuasca ceremony and 1 month and 1 year after. Secondary measures included ratings of altered states of consciousness and ego dissolution during the ayahuasca ceremony as well as global measures of mindfulness, satisfaction with life, depression, anxiety, and stress. Twenty participants completed baseline and 1-day follow-up, 19 completed measures at 1-month follow-up, and 17 completed measures at 1-year follow-up. BDI scores reduced from baseline (M = 22.7) to all post-ceremony measures (Ms 11.45, 12.89, and 8.88, for 1-day, 1-month, and 1-year follow-up, respectively). After 1 day, 12/20 participants were in remission (BDI < 13). Remission rates after 1 month and 1 year were 13/19 and 12/17, respectively. Three participants remained mildly depressed (BDI 14-19) at the 1-month and 1-year follow-up. Two participants did not respond and remained at a moderate/severe level of depression at 1-year follow-up. Reductions on the secondary mental health measures and increases in mindfulness and satisfaction with life were found up to 1 year post-ceremony. Improvements in clinical depression and mental health correlated with levels of experienced ego dissolution and oceanic boundlessness during the ceremony up to 1 month after the ceremony. Engagement in additional mental health treatments or use of another psychedelic during study participation may have contributed to improved mental health ratings at 1-year follow-up. Ayahuasca produces long-term mental health improvements in clinically depressed patients, which highlights its therapeutic potential.


Down-regulation of RFL, the FLO/LFY homolog of rice, accompanied with panicle branch initiation.
FLORICAULA (FLO) of Antirrhinum and LEAFY (FLY) of Arabidopsis regulate the formation of floral meristems. To examine whether same mechanisms control floral development in distantly related species such as grasses, we isolated RFL, FLO-LFY homolog of rice, and examined its expression and function. Northern analysis showed that RFL is expressed predominantly in very young panicle but not in mature florets, mature leaves, or roots. In situ hybridization revealed that RFL RNA was expressed in epidermal cells in young leaves at vegetative growth stage. After the transition to reproductive stage, RFL RNA was detected in all layers of very young panicle including the apical meristem, but absent in the incipient primary branches. As development of branches proceeds, RFL RNA accumulation localized in the developing branches except for the apical meristems of the branches and secondary branch primordia. Expression pattern of RFL raised a possibility that, unlike FLO and LFY, RFL might be involved in panicle branching. Transgenic Arabidopsis plants constitutively expressing RFL from the cauliflower mosaic virus 35S promoter were produced to test whether 35S-RFL would cause similar phenotype as observed in 35S-LFY plants. In 35S-RFL plants, transformation of inflorescence meristem to floral meristem was rarely observed. Instead, development of cotyledons, rosette leaves, petals, and stamens was severely affected, demonstrating that RFL function is distinct from that of LFY. Our results suggest that mechanisms controlling floral development in rice might be diverged from that of Arabidopsis and Antirrhinum.


Agonist and inverse agonist actions of beta-blockers at the human beta 2-adrenoceptor provide evidence for agonist-directed signaling.
Beta-blockers have beneficial effects in heart failure, although the underlying mechanism is unknown. Beta2-adrenoceptors, however, are proportionally higher in the failing human heart. This study shows several clinically used beta-blockers are agonists at the human beta2-adrenoceptor. Although these agonist effects were small at the cAMP level, they were substantial at the level of cAMP response element (CRE)-mediated gene transcription. Some of the effects of "beta-blockers" seen in heart failure may be related to the beta2-agonist actions of these compounds. CRE-gene transcription responses to beta2-agonists, forskolin, and cAMP-analogs were sensitive to p42/44-mitogen-activated protein (MAP) kinase pathway inhibitors. p42/44-MAP kinase activation was also shown directly by western blotting and enzyme-linked immunosorbent assay techniques. N-[2-(4-bromocinnamylamino)ethyl]-5-isoquinoline (H89; a protein kinase A inhibitor) stimulated cAMP accumulation and CRE gene transcription via the beta2-adrenoceptor at concentrations at which protein kinase A was inhibited, providing evidence for an alternative pathway. Propranolol, however, produced paradoxical effects; it reduced basal cAMP accumulation (via beta2-mediated inverse agonism) but stimulated beta2-mediated CRE gene transcription. This cannot be explained by a sequential pathway from Gs-adenylyl cyclase-cAMP to CRE binding protein phosphorylation. Both responses to propranolol were insensitive to pertussis toxin, thus excluding Gi-protein involvement. Propranolol CRE gene transcription responses were attenuated by p42/44-MAP kinase inhibitors and propranolol was also found to directly stimulate the p42/44-MAP kinase pathway. Studies of inositol phosphate accumulation and of protein kinase C or Rho kinase inhibitors on CRE-gene transcription provided no evidence for Gq/11 or G12/13 involvement. These data suggest that propranolol can simultaneously act as an inverse agonist through a Gs-coupled mechanism while stimulating the p42/44-MAP kinase pathway through an alternative G-protein-independent mechanism.


Synergistic effect of aqueous extract of Telfaria occidentalis on the biological activities of artesunate in Plasmodium berghei infected mice.
Resistance to most antimalarial drugs has encouraged the use of herbal preparations along with prescribed orthodox drugs. To investigate effect of co-administration of aqueous extract of T. occidentalis leaves; commonly used as antimalarial and haematinic agent in Nigeria and artesunate using P. berghei animal model. In vivo curative antiplasmodial effect of T. occidentalis (200mg/kg) alone and combination with artesunate (2mg/kg) were evaluated using albino mice infected with 10(6) parasitized erythrocytes of P. berghei intraperitoneally. The haematological parameters: haemoglobin level, red blood cells and white blood cells and packed cell volume were monitored using standard methods. Aqueous extract of T. occidentalis, artesunate and the combination gave 72.17±4.07%, 70.43± 4.27% and 85.43±3.65% reduction in parasitaemia after 48hours respectively. A significant enhancement of the PCV was obtained with the coadministration of artesunate and aqueous extract (p< 0.01). Similar trends were also observed with heamatological parameters at 72hours of administration. This study revealed a synergistic effect of the co-administration on parasite clearance rate of P. berghei infection in mice, with a significant enhancement of haematological parameters within 48 hours of administration. This indicates a rapid rate of recovery from plasmodial infections with the co-administration.


Childhood cancer near German nuclear power stations.
In 2008, the Kinderkrebs in der Umgebung von Kernkraftwerken (KiKK) study in Germany reported a 60% increase in solid cancers and a 120% increase in leukemias among children living within 5 km of all German nuclear power stations. The study has triggered debates as to the cause(s) of these increased cancers. This article reports on the findings of the KiKK study; discusses past and more recent epidemiological studies of leukemias near nuclear installations around the world, and outlines a possible biological mechanism to explain the increased cancers. This suggests that the observed high rates of infant leukemias may be a teratogenic effect from radionuclides incorporated by pregnant women living near nuclear reactors. Doses and risks from environmental emissions to embryos and fetuses may be larger than suspected. Hematopoietic tissues appear to be considerably more radiosensitive in embryos/fetuses than in newborn babies. Recommendations for advice to local residents and for further research are made.


Nutritional evaluation of pea (Pisum sativum L.) protein diets after mild hydrothermal treatment and with and without added phytase.
The effect of mild hydrothermal treatment and the addition of phytase under optimal conditions (pH 5.5, 37 degrees C) on the nutritive utilization of the protein of pea (Pisum sativum L.) flour was studied in growing rats by examining the chemical and biological balance. Mild hydrothermal treatment produced reductions of 83, 78, and 72%, respectively, in the levels of alpha-galactosides, phytic acid, and trypsin inhibitors and also produced a significant increase in the digestive utilization of protein. The additional fall in the levels of phytic acid caused by the addition of phytase did not lead to a subsequent improvement in the digestive utilization of protein. The mild hydrothermal treatment of pea flour produced a significant increase in the metabolic utilization of protein and carbohydrates, which was reflected in the protein efficiency ratio and food transformation growth indices. These effects were not observed in the phytase-supplemented pea diet.


Quantification of breast tumor microvascular permeability with feruglose-enhanced MR imaging: initial phase II multicenter trial.
To investigate use of the macromolecular contrast agent feruglose for differentiating and grading of human benign and malignant breast tumors on the basis of their microvascular characteristics. Sixty-three women with 63 primary breast lesions were examined with dynamic T1-weighted gradient-echo magnetic resonance (MR) imaging after intravenous injection of feruglose. A two-compartment unidirectional kinetic model applied to the dynamic data yielded estimates of the endothelial transfer coefficient, KPS, and the fractional plasma volume of the tumors. These MR imaging-derived parameters were correlated with the histologic tumor grade and quantified according to the Scarff-Bloom-Richardson (SBR) score by means of Pearson product moment correlation analyses. Differences between malignant and nonmalignant breast lesions with respect to KPS for feruglose were evaluated by means of the chi2 test and by calculating the sensitivity, specificity, and positive and negative predictive values. Histologic analysis revealed 26 benign and 37 malignant tumors. A moderate yet statistically significant correlation between KPS and SBR score was found (R = 0.496, P <.001). No significant correlation was observed between fractional plasma volume and SBR score (R = 0.085, P =.507). The KPS values were zero for 19 (73%) of the 26 benign tumors and were greater than zero for 27 (73%) of the 37 carcinomas. This distribution was significantly different (chi2 = 13.035, P =.001). With the criterion KPS > 0 in carcinomas, sensitivity was 0.73, specificity was 0.73, and the positive predictive value was 0.79. Quantitative measures of tumor microvascular permeability can be used for breast tumor characterization. The probability of breast tumor microvascular hyperpermeability to be associated with malignancy is 79%.


Cyclosporine metabolism and pharmacokinetics following intravenous and oral administration in the dog.
The influence of assay method on single dose cyclosporine (CsA) pharmacokinetics was studied in nine dogs receiving either i.v. or oral CsA. Samples were drawn from hepatic, portal, and systemic veins at various times after the dose and CsA levels were determined by radioimmunoassay (RIA) and high-performance liquid chromatography (HPLC). Blood concentration-time data were analyzed by nonlinear least-squares regression, using two-compartment models. RIA/HPLC ratios for all samples were greater than one, and did not change significantly over time. The mean RIA/HPLC ratios for samples drawn from all three veins were higher after oral than i.v. doses of the drug (P less than 0.05). Area under the concentration-time curve (AUC) was higher and systemic clearance (CIs) lower than calculated on the basis of RIA results, regardless of the route of administration. AUC calculated for CsA metabolites (RIA-HPLC) was highest in the portal vein after an oral dose of CsA. Bioavailability was 20.4% and 27.0% when estimated using HPLC and RIA data, respectively. The mean CsA metabolite index (CMI), when calculated for hepatic, portal, or systemic vein, was greater when the drug was administered orally. The mean hepatic extraction ratio (HER) of the parent drug and for CsA metabolites was approximately 23% in i.v. and p.o. studies. These results suggest that the gastrointestinal tract may play a role in the metabolism of CsA when the drug is administered orally. In addition, if CsA metabolites not measured by HPLC have either toxic or immunosuppressive properties, the RIA assay may be more useful for monitoring patients.


Cost Effectiveness of a Sugar-Sweetened Beverage Excise Tax in the U.S.
Reducing sugar-sweetened beverage consumption through taxation is a promising public health response to the obesity epidemic in the U.S. This study quantifies the expected health and economic benefits of a national sugar-sweetened beverage excise tax of $0.01/ounce over 10 years. A cohort model was used to simulate the impact of the tax on BMI. Assuming ongoing implementation and effect maintenance, quality-adjusted life-years gained and disability-adjusted life-years and healthcare costs averted were estimated over the 2015-2025 period for the 2015 U.S. Costs and health gains were discounted at 3% annually. Data were analyzed in 2014. Implementing the tax nationally would cost $51 million in the first year. The tax would reduce sugar-sweetened beverage consumption by 20% and mean BMI by 0.16 (95% uncertainty interval [UI]=0.06, 0.37) units among youth and 0.08 (95% UI=0.03, 0.20) units among adults in the second year for a cost of $3.16 (95% UI=$1.24, $8.14) per BMI unit reduced. From 2015 to 2025, the policy would avert 101,000 disability-adjusted life-years (95% UI=34,800, 249,000); gain 871,000 quality-adjusted life-years (95% UI=342,000, 2,030,000); and result in $23.6 billion (95% UI=$9.33 billion, $54.9 billion) in healthcare cost savings. The tax would generate $12.5 billion in annual revenue (95% UI=$8.92, billion, $14.1 billion). The proposed tax could substantially reduce BMI and healthcare expenditures and increase healthy life expectancy. Concerns regarding the potentially regressive tax may be addressed by reduced obesity disparities and progressive earmarking of tax revenue for health promotion.


Emergence of fluoroquinolone resistance in Campylobacter jejuni and Campylobacter coli in subjects from Finland.
The in vitro susceptibilities of 102 human campylobacter strains isolated between 1978 and 1980 and 100 strains isolated in 1990 to ciprofloxacin, norfloxacin, erythromycin, gentamicin, and doxycycline were examined. The biotypes and heat-stable serotypes of the strains as well as antimicrobial treatments and travel history of the campylobacter-positive patients were also studied. The results indicated that susceptibility to erythromycin, gentamicin, and doxycycline has remained the same during the past 10 years. No gentamicin-resistant strains were found. Resistance to erythromycin was 3% in both groups of strains. However, the number of norfloxacin-resistant strains increased from 4 to 11% in the follow-up period, and ciprofloxacin-resistant strains, which had not occurred 10 years ago, composed 9% of the strains isolated in 1990. Thus, the increase of fluoroquinolone resistance in Campylobacter jejuni and Campylobacter coli has been significant in Finland in the past 10 years.


Methylphenidate use among medical students at Ben-Gurion University of the Negev.
Methylphenidate is a psychotropic agent commonly used for the treatment of attention deficit disorder with or without hyperactivity and narcolepsy in children and adults. The awareness to attention deficit disorder as well as the non-medical use of methylphenidate for cognitive enhancement has increased during the past years. To evaluate the medical and non-medical use of methylphenidate among medical students in the Ben-Gurion University of the Negev. Medical students were asked to report methylphenidate use, symptoms and diagnosis of attention deficit disorder using a structured questionnaire. A total of 229 students participated in the study, out of which 105 (45.9%) were in the pre-clinical years of medical school. Twenty-two students (9.6%) were previously diagnosed with attention deficit disorder. Lifelong use of methylphenidate was reported by 39 (17%) students, while 31 students (13.5%) reported using methylphenidate during the preceding 12 month. In the beginning of medical school, only 7% of the students used methylphenidate, most of them began using it during pre-clinical academic years. High rates of attention deficit disorder compared to the general population were reported by medical students. The rate of methylphenidate use is similar to recent report from a US medical school, and is considerably higher than in college students population. Many medical students are using methylphenidate without a medical indication. Further study is needed to evaluate the effect of methylphenidate on academic performance of healthy adults.


Digital biomarkers of anxiety disorder symptom changes: Personalized deep learning models using smartphone sensors accurately predict anxiety symptoms from ecological momentary assessments.
Smartphones are capable of passively capturing persons' social interactions, movement patterns, physiological activation, and physical environment. Nevertheless, little research has examined whether momentary anxiety symptoms can be accurately assessed using these methodologies. In this research, we utilize smartphone sensors and personalized deep learning models to predict future anxiety symptoms among a sample reporting clinical anxiety disorder symptoms. Participants (N = 32) with generalized anxiety disorder and/or social anxiety disorder (based on self-report) installed a smartphone application and completed ecological momentary assessment symptoms assessing their anxiety and avoidance symptoms hourly for the course of one week (T = 2007 assessments). During the same period, the smartphone app collected information about physiological activation (heart rate and heart rate variability), exposure to light, social contact, and GPS location. GPS locations were coded to reveal the type of location and the weather information. Personalized deep learning models using the smartphone sensor data were capable of predicting the majority of total variation in anxiety symptoms (R2 = 0.748) and predicting a large proportion of within-person variation at the hour-by-hour level (mean R2 = 0.385). These results suggest that personalized deep learning models using smartphone sensor data are capable of accurately predicting future anxiety disorder symptom changes.


Maternal serum interleukin-1 beta, -6 and -8 levels and potential determinants in pregnancy and peripartum.
To measure maternal serum interleukins (IL) in pregnancy, delivery and early puerperium, and to identify their potential determinants. Prospective longitudinal measures of serum IL-1 beta, IL-6 and IL-8 in 38 healthy pregnant women at antenatal visits, through labor and delivery, with clinical correlates (infection, vaginal hemorrhage and anemia) recorded by questionnaire. Pregnancy IL levels remained consistently low. IL-1 beta increased shortly before delivery, then returned to pregnant levels, except where blood loss exceeded 500 ml. IL-6 and IL-8 rose at labor onset and exceeded pregnancy levels through postpartum day three. Postpartum IL-6 was higher after non-elective cesarean section than after spontaneous delivery (P < 0.0001), and where blood loss exceeded 500 ml. IL-6 and IL-8 were higher with systemic infection during delivery (P < 0.0001) and on postpartum day one (P < 0.05); IL-8 was higher in anemia (delivery: P < 0.005; postpartum day 1: P < 0.05). Differences due to delivery mode and systemic infection remained significant after correction for other conditions. Labor-dependent inflammation increases all IL levels at delivery. Further studies with larger sample sizes are required to establish reference values differentiating physiology from pathology as an aid to peripartum management.


Appendiceal foreign body in an infant.
Many children are hospitalized because of foreign body ingestion. In such circumstances, the gastroenterologist must consider the timing of ingestion; the size, type, and location of the object ingested; and the patient's symptoms. But appendiceal foreign body in infant is very rare. A 12-month-old boy visited because of swallowing small objects 2 weeks ago. Three small beads were found in the right lower quadrant on abdominal radiography. A 7-year-old boy was admitted for treatment of appendiceal foreign body, which was detected by accident 3 months ago. A 7-year-old boy had abdominal pain and underwent immediate appendectomy. However, the 12-month-old child was asymptomatic, which led to deliberation in regard to wait time for an appendectomy in younger children with asymptomatic appendiceal foreign body. He underwent prophylactic appendectomy after 2 months. An appendiceal foreign body is very rare in infant and there are currently no treatment guidelines. We report 2 cases of appendiceal foreign body including infant who gave us difficult decisions.


Overt and subclinical hypothyroidism: who to treat and how.
Hypothyroidism denotes deficient production of thyroid hormone by the thyroid gland and can be primary (abnormality in thyroid gland itself) or secondary/central (as a result of hypothalamic or pituitary disease). The term 'subclinical hypothyroidism' is used to define that grade of primary hypothyroidism in which there is an elevated thyroid-stimulating hormone (TSH) concentration in the presence of normal serum free thyroxine (T4) and triiodothyronine (T3) concentrations. Subclinical hypothyroidism may progress to overt hypothyroidism in approximately 2-5% cases annually. All patients with overt hypothyroidism and subclinical hypothyroidism with TSH >10 mIU/L should be treated. There is consensus on the need to treat subclinical hypothyroidism of any magnitude in pregnant women and women who are contemplating pregnancy, to decrease the risk of pregnancy complications and impaired cognitive development of the offspring. However, controversy remains regarding treatment of non-pregnant adult patients with subclinical hypothyroidism and serum TSH values ≤10 mIU/L. In this subgroup, treatment should be considered in symptomatic patients, patients with infertility, and patients with goitre or positive anti-thyroid peroxidase (TPO) antibodies. Limited evidence suggests that treatment of subclinical hypothyroidism in patients with serum TSH of up to 10 mIU/L should probably be avoided in those aged >85 years. Other pituitary hormones should be evaluated in patients with central hypothyroidism, especially assessment of the hypothalamic-pituitary-adrenal axis, since hypocortisolism, if present, needs to be rectified prior to initiating thyroid hormone replacement. Levothyroxine (LT4) monotherapy remains the current standard for management of primary, as well as central, hypothyroidism. Treatment can be started with the full calculated dose for most young patients. However, treatment should be initiated at a low dose in elderly patients, patients with coronary artery disease and patients with long-standing severe hypothyroidism. In primary hypothyroidism, treatment is monitored with serum TSH, with a target of 0.5-2.0 mIU/L. In patients with central hypothyroidism, treatment is tailored according to free or total T4 levels, which should be maintained in the upper half of the normal range for age. In patients with persistently elevated TSH despite an apparently adequate replacement dose of LT4, poor compliance, malabsorption and the presence of drug interactions should be checked. Over-replacement is common in clinical practice and is associated with increased risk of atrial fibrillation and osteoporosis, and hence should be avoided.


Carbohydrate fractions from cooked fish promote iron uptake by Caco-2 cells.
The objective of this study was to isolate and characterize the meat factor(s) that enhances nonheme iron bioavailability using various analytical and in vitro cell culture techniques. Nonheme iron bioavailability was measured via radiolabeled iron uptake or ferritin formation in Caco-2 cells. Fish haddock fillet was cooked and lyophilized to be used as the muscle tissue of choice because of its low intrinsic iron content. It was demonstrated that the low pH of the stomach (pH 2.0) was the primary factor responsible for initiating the enhancing effect of fish on nonheme iron uptake. Subsequently, cooked fish samples were titrated with HCl to pH 2.0 and incubated for 1 h without digestive enzymes to release the factor(s) from the fish. The supernatant of this acidic digest was then used as a starting material for the meat factor isolation procedures. Fractions generated through Sephadex G-25 size exclusion increased Caco-2 cell iron uptake approximately 9-fold. Subsequent chromatography of these fractions via C18 reverse-phase HPLC were conducted, and enhancing activity was observed only in the "injection peak." This observation coupled with protein measurement and amino acid composition analysis revealed that the active fractions contained negligible amounts of proteins or amino acids. Active fractions were highly enriched with carbohydrates. Subsequent chromatography via high performance anion exchange chromatography with pulsed amperometric detection yielded 3 active peaks that increased Caco-2 cell iron uptake 3.4- to 4.9-fold. Our results indicate that specific carbohydrates contribute to the enhancing effect of meat on iron uptake by the enterocyte. These carbohydrates may be oligosaccharides originating from glycosaminoglycans in the extracellular matrix of muscle tissue.


Increase in Trauma Leading to Psychological Injury Among Canadian Homelessness Services Workers During the COVID-19 Pandemic.
This study examined the traumatic psychological impact of the pandemic on frontline workers in homelessness services. Staff from homelessness serving organizations completed pre- and mid-COVID pandemic surveys measuring traumatic stress symptoms (PTSS), burnout, and job-related traumatic experiences. The mid-pandemic survey was expanded to seven Canadian cities to determine prevalence of workplace PTSS nationally. In the comparison group, baseline rates of PTSS (41%) rose to 47.3% (N = 164), while 75% reported low-moderate levels of burnout both times. Nationwide, PYSS was 51% (N = 574) Case managers working at remotely had greater levels of PTSS. COVID-19 exacerbated risk of psychological workplace injury from traumatic stress, however, burnout did not increase significantly, indicating the primary dynamic as anxiety and emotional exhaustion associated with ubiquitous trauma induced by COVID-19. Working remotely increased the hazards of psychological workplace injury.


Discriminative stimulus effects of the nonpeptidic delta-opioid agonist SNC80 in rhesus monkeys.
Five rhesus monkeys were trained to discriminate the nonpeptidic, delta-opioid agonist SNC80 (0.32 mg/kg i.m.) from saline by using a food-reinforced drug-discrimination procedure. Cumulative doses of SNC80 produced a dose-dependent increase in SNC80-appropriate responding and a dose-dependent decrease in response rate. In time-course studies, peak effects of the training dose of SNC80 were observed after 15 min, and these effects diminished over 240 min. In substitution studies, other piperazinyl benzamide delta agonists (SNC86, SNC162, and SNC243A) substituted for SNC80 with relative potencies similar those of SNC80. However, SNC67, the (-)-enantiomer of SNC80, did not occasion SNC80-appropriate responding up to a dose (32.0 mg/kg) that produced convulsions in one monkey. The mu agonists morphine and fentanyl and the kappa agonists U-50,488 and enadoline failed to substitute for SNC80 up to doses that eliminated responding. Two nonopioids (the N-methyl-D-aspartate antagonist ketamine and the monoamine reuptake inhibitor cocaine) also produced primarily saline-appropriate responding. Both the discriminative stimulus and rate-decreasing effects of SNC80 were antagonized by the delta-selective antagonist naltrindole (0.01-1.0 mg/kg) but not by doses of the opioid antagonist quadazocine (0.1-1.0 mg/kg) that block the effects of mu and kappa agonists. These data suggest that the discriminative stimulus effects of SNC80 are mediated by delta-opioid receptors and that the discriminative stimulus effects of delta opioids in primates can be differentiated from the effects of other opioid and nonopioid compounds.


A theory of alternative designs for biochemical control systems.
Any theory of alternative designs must include (a) an appropriate language for describing alternatives, (b) methods of analysis for relating system behavior to changes in design elements, and (c) methods for critically comparing the behavior of alternatives. Over the past two decades we have been reasonably successful in developing a theory of alternative designs that meets these basic requirements. The language for describing alternatives is provided by the S-system of differential equations discussed elsewhere in this volume. The methods of analysis consist of adaptations of conventional methods from control theory and newly developed algorithms specific for S-systems. The methods for making critical comparisons perhaps are less familiar and will be treated in some detail. Several applications of this theory to alternative designs for biochemical control are reviewed.


Toxicity of manufactured zinc oxide nanoparticles in the nematode Caenorhabditis elegans.
Information describing the possible impacts of manufactured nanoparticles on human health and ecological receptors is limited. The objective of the present study was to evaluate the potential toxicological effects of manufactured zinc oxide nanoparticles (ZnO-NPs; 1.5 nm) compared to aqueous zinc chloride (ZnCl2) in the free-living nematode Caenorhabditis elegans. Toxicity of both types of Zn was investigated using the ecologically relevant endpoints of lethality, behavior, reproduction, and transgene expression in a mtl-2::GFP (gene encoding green fluorescence protein fused onto the metallothionein-2 gene promoter) transgenic strain of C. elegans. Zinc oxide nanoparticles showed no significant difference from ZnCl2 regarding either lethality or reproduction in C. elegans, as indicated by their median lethal concentrations (LC50s; p = 0.29, n=3) and median effective concentrations (EC50s; Z = 0.835, p = 0.797). Also, no significant difference was found in EC50s for behavioral change between ZnO-NPs (635 mg Zn/L; 95% confidence interval [CI], 477-844 mg Zn/L) and ZnCl2 (546 mg Zn/L; 95% CI, 447-666 mg Zn/L) (Z = 0.907, p = 0.834). Zinc oxide nanoparticles induced transgene expression in the mtl-2::GFP transgenic C. elegans in a manner similar to that of ZnCl2, suggesting that intracellular biotransformation of the nanoparticles might have occurred or the nanoparticles have dissolved to Zn2+ to enact toxicity. These findings demonstrate that manufactured ZnO-NPs have toxicity to the nematode C. elegans similar to that of aqueous ZnCl2.


Surgeon case volume, not institution case volume, is the primary determinant of in-hospital mortality after elective open abdominal aortic aneurysm repair.
Studies analyzing the effects of volume on outcomes after abdominal aortic aneurysm (AAA) repair have primarily centered on institutional volume and not on individual surgeon volume. We sought to determine the relative effects of both surgeon and institution volume on mortality after open and endovascular aneurysm repair (EVAR) for intact AAAs. The Nationwide Inpatient Sample (2003-2007) was queried to identify all patients undergoing open repair and EVAR for nonruptured AAAs. To calculate surgeon and institution volume, 11 participating states that record a unique physician identifier for each procedure were included. Surgeon and institution volume were defined as low (first quintile), medium (second, third, or fourth quintile), and high (fifth quintile). Stratification by institution volume and then by surgeon volume was performed to analyze the primary endpoint: in-hospital mortality. Multivariable models were used to evaluate the association of institution and surgeon volume with mortality for open repair and EVAR, controlling for potential confounders. During the study period, 5972 open repairs and 8121 EVARs were performed. For open AAA repair, a significant mortality reduction was associated with both annual institution volume (low <7, medium 7-30, and high >30) and surgeon volume (low ≤ 2, medium 3-9, and high >9). High surgeon volume conferred a greater mortality reduction than did high institution volume. When low and medium volume institutions were stratified by surgeon volume, mortality after open AAA repair was inversely proportional to surgeon volume (8.7%, 3.6%, and 0%; P < .0001, for low, medium, and high-volume surgeons at low-volume institutions; and 6.7%, 4.8%, and 3.3%; P = .02, for low, medium, and high-volume surgeons at medium-volume institutions). High-volume institutions stratified by surgeon volume demonstrated the same trend (5.1%, 3.4%, and 2.8%), but this finding was not statistically significant (P = .57). Multivariable analysis was confirmatory: low surgeon volume independently predicted mortality (odds ratio [OR], 2.0; 95% confidence interval [CI], 1.3-3.1; P < .001); low institution volume did not (P = .1). For EVAR, neither institution volume nor surgeon volume influenced mortality (univariate or multivariable). The primary factor driving the mortality reduction associated with case volume after open AAA repair is surgeon volume, not institution volume. Regionalization of AAAs should focus on open repair, as EVAR outcomes are equivalent across volume levels. Payers may need to re-evaluate strategies that encourage open AAA repair at high-volume institutions if specific surgeon volume is not considered.


Acute posttraumatic renal failure: a multicenter perspective.
Acute renal failure (ARF) following trauma is rare. Historically, ARF has been associated with a high mortality rate. To investigate this entity we conducted a retrospective review of 72,757 admissions treated at nine regional trauma centers over a 5-year period. Seventy-eight patients (0.098%) developed acute renal failure requiring hemodialysis. Detailed demographic, clinical, and outcome data were collected. Patients with pre-existing medical conditions (group I) had a 70% increase in mortality over those without pre-existing conditions (p less than 0.004). Twenty-four patients (31%) developed ARF less than 6 days after injury (group II). The remainder (group III) developed late renal failure (mean time to first dialysis, 23 days). The predominant cause of death was multiple organ failure (82%). There were no differences in mortality because of multiple organ failure among the three groups of patients. Of the 33 survivors, six (18%) were discharged with renal insufficiency, three (9%) were discharged on dialysis, 23 (70%) were discharged home or to rehabilitation, and 27 (82%) had no significant evidence of renal insufficiency. Posttraumatic renal failure requiring hemodialysis is rare (incidence, 107 per 100,000 trauma center admissions), but the mortality rate remains high (57%). Two thirds of the cases of posttraumatic renal failure develop late and are secondary to multiple organ failure; one third of the cases of posttraumatic renal failure develop early and may result from inadequate resuscitation.


Specificity of the helper T cell for the cytolytic T lymphocyte response to influenza viruses.
We have described a model system in which helper T cells are required to mount a primary antiviral cytolytic T lymphocyte response. The radioresistant helper cell can be found in the spleens of mice that have been immunized subcutaneously with influenza viruses 6-8 d previously. These helper cells appear to be type specific but cross-reactive among the subtypes of influenza A viruses. The phenotypes of the interacting cell populations were determined.


Systematic analysis of severity in a widely used cognitive depression model for mice.
Animal models in psychiatric research are indispensable for insights into mechanisms of behaviour and mental disorders. Distress is an important aetiological factor in psychiatric diseases, especially depression, and is often used to mimic the human condition. Modern bioethics requires balancing scientific progress with animal welfare concerns. Therefore, scientifically based severity assessment of procedures is a prerequisite for choosing the least compromising paradigm according to the 3Rs principle. Evidence-based severity assessment in psychiatric animal models is scarce, particularly in depression research. Here, we assessed severity in a cognitive depression model by analysing indicators of stress and well-being, including physiological (body weight and corticosterone metabolite concentrations) and behavioural (nesting and burrowing behaviour) parameters. Additionally, a novel approach for objective individualised severity grading was employed using clustering of voluntary wheel running (VWR) behaviour. Exposure to the paradigm evoked a transient elevation of corticosterone, but neither affected body weight, nesting or burrowing behaviour. However, the performance in VWR was impaired after recurrent stress exposure, and the individual severity level increased, indicating that this method is more sensitive in detecting compromised welfare. Interestingly, the direct comparison to a somatic, chemically induced colitis model indicates less distress in the depression model. Further objective severity assessment studies are needed to classify the severity of psychiatric animal models in order to balance validity and welfare, reduce the stress load and thus promote refinement.


A global study of forensically significant calliphorids: implications for identification.
A proliferation of molecular studies of the forensically significant Calliphoridae in the last decade has seen molecule-based identification of immature and damaged specimens become a routine complement to traditional morphological identification as a preliminary to the accurate estimation of post-mortem intervals (PMI), which depends on the use of species-specific developmental data. Published molecular studies have tended to focus on generating data for geographically localised communities of species of importance, which has limited the consideration of intraspecific variation in species of global distribution. This study used phylogenetic analysis to assess the species status of 27 forensically important calliphorid species based on 1167 base pairs of the COI gene of 119 specimens from 22 countries, and confirmed the utility of the COI gene in identifying most species. The species Lucilia cuprina, Chrysomya megacephala, Ch. saffranea, Ch. albifrontalis and Calliphora stygia were unable to be monophyletically resolved based on these data. Identification of phylogenetically young species will require a faster-evolving molecular marker, but most species could be unambiguously characterised by sampling relatively few conspecific individuals if they were from distant localities. Intraspecific geographical variation was observed within Ch. rufifacies and L. cuprina, and is discussed with reference to unrecognised species.


Experimental Approaches for Characterizing the Endocrine-Disrupting Effects of Environmental Chemicals in Fish.
Increasing industrial and agricultural activities have led to a disturbing increase of pollutant discharges into the environment. Most of these pollutants can induce short-term, sustained or delayed impacts on developmental, physiological, and behavioral processes that are often regulated by the endocrine system in vertebrates, including fish, thus they are termed endocrine-disrupting chemicals (EDCs). Physiological impacts resulting from the exposure of these vertebrates to EDCs include abnormalities in growth and reproductive development, as many of the prevalent chemicals are capable of binding the receptors to sex steroid hormones. The approaches employed to investigate the action and impact of EDCs is largely dependent on the specific life history and habitat of each species, and the type of chemical that organisms are exposed to. Aquatic vertebrates, such as fish, are among the first organisms to be affected by waterborne EDCs, an attribute that has justified their wide-spread use as sentinel species. Many fish species are exposed to these chemicals in the wild, for either short or prolonged periods as larvae, adults, or both, thus, studies are typically designed to focus on either acute or chronic exposure at distinct developmental stages. The aim of this review is to provide an overview of the approaches and experimental methods commonly used to characterize the effects of some of the environmentally prevalent and emerging EDCs, including 17 α-ethinylestradiol, nonylphenol, BPA, phthalates, and arsenic; and the pervasive and potential carriers of EDCs, microplastics, on reproduction and growth. In vivo and in vitro studies are designed and employed to elucidate the direct effects of EDCs at the organismal and cellular levels, respectively. In silico approaches, on the other hand, comprise computational methods that have been more recently applied with the potential to replace extensive in vitro screening of EDCs. These approaches are discussed in light of model species, age and duration of EDC exposure.


Daily lives of residents with dementia in nursing homes: development of the Maastricht electronic daily life observation tool.
Daily life is a dynamic and multidimensional concept, for which appropriate assessment tools are lacking. This study describes the development of the Maastricht Electronic Daily Life Observation tool (MEDLO-tool), a freely accessible, easy to use, electronic observation tool to assess relevant daily life aspects for nursing home residents with dementia. (1) Determining relevant aspects of daily life for nursing home residents with dementia based on a literature search and expert interviews; (2) pilot testing observation procedures and operationalizations of the aspects of daily life; and (3) exploring inter-rater reliability and feasibility of the tool in a nursing home facility with 16 residents (56% female, mean age: 77). The following aspects of daily life are assessed with the MEDLO-tool: (1) activity (activity performed by resident, engagement in this activity, and the degree of physical effort); (2) physical environment (location of the resident and interaction with the physical environment); (3) social interaction (the level and type of social interaction, and with whom this social interaction took place); and (4) emotional well-being (mood and agitation). Each aspect of daily life is observed and scored using standardized scoring options. Agreement on the aspects is high with an average absolute agreement of 86%. Users of the MEDLO-tool indicated that it was feasible in practice and contained clear operationalization of the aspects of daily life. The MEDLO-tool is a promising tool to gain real time insight into the aspects of the daily lives of nursing home residents with dementia.


An evidence-based approach to prostate cancer follow-up.
The follow-up required for patients with prostate cancer is critically dependent upon the stage of disease and the ultimate goal for treatment. A major difficulty in follow-up in prostate cancer is the lack of data on outcome of various treatment modalities. Additionally, there is a lack of data on the use of treatment modalities early in the course of prostate cancer. Despite these limitations, there is a need to develop an approach to follow these patients pending further study. In this review, we critically assess the natural course of untreated prostate cancer, the complications of local therapy, and the controversy over early versus delayed hormonal therapy. As a result of this discussion, common themes emerge. Most patients diagnosed with prostate cancer die of causes other than prostate cancer such as cardiovascular disease and therefore require additional follow-up. Since patients experience local problems such as urinary obstruction more commonly than symptomatic metastatic disease, instruments to assess urinary symptoms are discussed. Finally, follow-up as a means to determine eligibility for clinical studies is discussed.


Identification and characterization of aortic valve mesenchymal progenitor cells with robust osteogenic calcification potential.
Advanced valvular lesions often contain ectopic mesenchymal tissues, which may be elaborated by an unidentified multipotent progenitor subpopulation within the valve interstitium. The identity, frequency, and differentiation potential of the putative progenitor subpopulation are unknown. The objectives of this study were to determine whether valve interstitial cells (VICs) contain a subpopulation of multipotent mesenchymal progenitor cells, to measure the frequencies of the mesenchymal progenitors and osteoprogenitors, and to characterize the osteoprogenitor subpopulation because of its potential role in calcific aortic valve disease. The multilineage potential of freshly isolated and subcultured porcine aortic VICs was tested in vitro. Progenitor frequencies and self-renewal capacity were determined by limiting dilution and colony-forming unit assays. VICs were inducible to osteogenic, adipogenic, chondrogenic, and myofibrogenic lineages. Osteogenic differentiation was also observed in situ in sclerotic porcine leaflets. Primary VICs had strikingly high frequencies of mesenchymal progenitors (48.0 +/- 5.7%) and osteoprogenitors (44.1 +/- 12.0%). High frequencies were maintained for up to six population doublings, but decreased after nine population doublings to 28.2 +/- 9.9% and 5.8 +/- 1.3%, for mesenchymal progenitors and osteoprogenitors, respectively. We further identified the putative osteoprogenitor subpopulation as morphologically distinct cells that occur at high frequency, self-renew, and elaborate bone matrix from single cells. These findings demonstrate that the aortic valve is rich in a mesenchyma l progenitor cell population that has strong potential to contribute to valve calcification.


Inhibition of Sirt1 promotes neural progenitors toward motoneuron differentiation from human embryonic stem cells.
Several protocols direct human embryonic stem cells (hESCs) toward differentiation into functional motoneurons, but the efficiency of motoneuron generation varies based on the human ESC line used. We aimed to develop a novel protocol to increase the formation of motoneurons from human ESCs. In this study, we tested a nuclear histone deacetylase protein, Sirt1, to promote neural precursor cell (NPC) development during differentiation of human ESCs into motoneurons. A specific inhibitor of Sirt1, nicotinamide, dramatically increased motoneuron formation. We found that about 60% of the cells from the total NPCs expressed HB9 and βIII-tubulin, commonly used motoneuronal markers found in neurons derived from ESCs following nicotinamide treatment. Motoneurons derived from ESC expressed choline acetyltransferase (ChAT), a positive marker of mature motoneuron. Moreover, we also examined the transcript levels of Mash1, Ngn2, and HB9 mRNA in the differentiated NPCs treated with the Sirt1 activator resveratrol (50 μM) or inhibitor nicotinamide (100 μM). The levels of Mash1, Ngn2, and HB9 mRNA were significantly increased after nicotinamide treatment compared with control groups, which used the traditional protocol. These results suggested that increasing Mash1 and Ngn2 levels by inhibiting Sirt1 could elevate HB9 expression, which promotes motoneuron differentiation. This study provides an alternative method for the production of transplantable motoneurons, a key requirement in the development of hESC-based cell therapy in motoneuron disease.


Sport for tall.
Eight new events (handball, basketball, and six rowing events) were introduced for women in the Olympic Games at Montreal in 1976. Of 187 women rowers who competed at Montreal, none was shorter than the mean height (162 cm, 64 in) of women aged 18-24 in the United States. In team events only two out of 250 participants were shorter than the reference mean. Even among the tall, it was the taller participants who won medals. What does the slogan "Sport for All" mean in this context? Moreover, the physical size required of champion rowers and basketball players is not to be found in some Asian, African, and Latin American populations. International contests in many such events therefore seem to be at variance with the first charter of the Olympic Games. An independent reviewing body is urgently needed to examine the merits of man made rules in many sporting contests.


Halothane hepatitis and the anaesthesiologist.
There is no longer any doubt that liver dysfunction following surgery and anaesthesia may sometimes be attributable to the inhalational anaesthetic agent halothane. Much controversy still surrounds the entity of halothane hepatitis, which continues to vex the practising anaesthesiologist. The incidence, causation, pathogenesis and diagnosis of halothane hepatitis are presented, and the safety and practicality of halothane administration in the presence of liver dysfunction are discussed.


The Na+-dependent chloride-bicarbonate exchanger SLC4A8 mediates an electroneutral Na+ reabsorption process in the renal cortical collecting ducts of mice.
Regulation of sodium balance is a critical factor in the maintenance of euvolemia, and dysregulation of renal sodium excretion results in disorders of altered intravascular volume, such as hypertension. The amiloride-sensitive epithelial sodium channel (ENaC) is thought to be the only mechanism for sodium transport in the cortical collecting duct (CCD) of the kidney. However, it has been found that much of the sodium absorption in the CCD is actually amiloride insensitive and sensitive to thiazide diuretics, which also block the Na-Cl cotransporter (NCC) located in the distal convoluted tubule. In this study, we have demonstrated the presence of electroneutral, amiloride-resistant, thiazide-sensitive, transepithelial NaCl absorption in mouse CCDs, which persists even with genetic disruption of ENaC. Furthermore, hydrochlorothiazide (HCTZ) increased excretion of Na+ and Cl- in mice devoid of the thiazide target NCC, suggesting that an additional mechanism might account for this effect. Studies on isolated CCDs suggested that the parallel action of the Na+-driven Cl-/HCO3- exchanger (NDCBE/SLC4A8) and the Na+-independent Cl-/HCO3- exchanger (pendrin/SLC26A4) accounted for the electroneutral thiazide-sensitive sodium transport. Furthermore, genetic ablation of SLC4A8 abolished thiazide-sensitive NaCl transport in the CCD. These studies establish what we believe to be a novel role for NDCBE in mediating substantial Na+ reabsorption in the CCD and suggest a role for this transporter in the regulation of fluid homeostasis in mice.


Oncogenic K-Ras decouples glucose and glutamine metabolism to support cancer cell growth.
Oncogenes such as K-ras mediate cellular and metabolic transformation during tumorigenesis. To analyze K-Ras-dependent metabolic alterations, we employed ¹³C metabolic flux analysis (MFA), non-targeted tracer fate detection (NTFD) of ¹⁵N-labeled glutamine, and transcriptomic profiling in mouse fibroblast and human carcinoma cell lines. Stable isotope-labeled glucose and glutamine tracers and computational determination of intracellular fluxes indicated that cells expressing oncogenic K-Ras exhibited enhanced glycolytic activity, decreased oxidative flux through the tricarboxylic acid (TCA) cycle, and increased utilization of glutamine for anabolic synthesis. Surprisingly, a non-canonical labeling of TCA cycle-associated metabolites was detected in both transformed cell lines. Transcriptional profiling detected elevated expression of several genes associated with glycolysis, glutamine metabolism, and nucleotide biosynthesis upon transformation with oncogenic K-Ras. Chemical perturbation of enzymes along these pathways further supports the decoupling of glycolysis and TCA metabolism, with glutamine supplying increased carbon to drive the TCA cycle. These results provide evidence for a role of oncogenic K-Ras in the metabolic reprogramming of cancer cells.


Advances in the production and handling of encoded microparticles.
Here we highlight emerging technologies in the synthesis, handling, and application of encoded microparticles for multiplexed assays. Traditionally, in drug discovery and life sciences research, multiple reactions will be conducted in parallel using microwell plate formats or microfluidic implementations, in which volumes are confined and reactions annotated by knowledge of what reagents were added to each volume. Microparticle-based information carriers provide an alternative approach to performing such multiplexed reactions, in which reactions and events are instead annotated with unique codes associated with the solid-phase particle. One challenge has been in creating a unique and large enough code set that is also easily readout, and we highlight two approaches that have brought orthogonal optical tagging techniques to bear. Another challenge has been that in such approaches, reactions have usually been confined to the surface of, or within the bulk of the specifically-tagged particle. We also highlight a creative approach and strategy for multiplexing - called "partipetting"- in which the coded particle can be a carrier of a unique fluid reagent.


Giant Cell Tumor of the Capitate Treated with Excision and Midcarpal Fusion.
Background Giant cell tumor (GCT) of bone is rare in the carpus, and only 11 cases have been reported in the capitate bone. The problem with this location is the high recurrence rate due to easy extension to the adjacent joint spaces and bones. We describe a case of GCT on the capitate bone and the treatment in comparison with the previously reported cases. Case Description The case report concerns a 48-year-old woman with a GCT of the capitate diagnosed on curettage. The treatment consisted of large resection with the lunatum and third metacarpal arthrodesis, with a successful union at 2 years follow-up, no recurrence, and an improved function of the wrist. Literature Review When treated by curettage (alone or with adjuvant procedures), the GCT of the capitate recurred in four out of five cases. All tumors treated with large resection did not recur. Clinical Relevance Recommended treatment of GCT of the capitate is resection with carpometacarpal arthrodesis.


Predictors of multiple suicide attempts among suicidal black adolescents.
Psychopathology, social support, and interpersonal orientation were studied in relation to suicide attempt status in acutely suicidal, psychiatrically hospitalized Black adolescents and a matched sample of White adolescents. In the total sample, multiple attempters were differentiated by lower perceived support. Within the Black youth subsample, social comparison and positive stimulation from others differentiated multiple attempters from single attempters/ideators. Only suicidal ideation predicted multiple attempts among White youth and only higher interpersonal orientation predicted multiple suicide attempts within Black adolescents.


B chromosomes of multiple species have intense evolutionary dynamics and accumulated genes related to important biological processes.
One of the biggest challenges in chromosome biology is to understand the occurrence and complex genetics of the extra, non-essential karyotype elements, commonly known as supernumerary or B chromosomes (Bs). The non-Mendelian inheritance and non-pairing abilities of B chromosomes make them an interesting model for genomics studies, thus bringing to bear different questions about their genetic composition, evolutionary survival, maintenance and functional role inside the cell. This study uncovers these phenomena in multiple species that we considered as representative organisms of both vertebrate and invertebrate models for B chromosome analysis. We sequenced the genomes of three animal species including two fishes Astyanax mexicanus and Astyanax correntinus, and a grasshopper Abracris flavolineata, each with and without Bs, and identified their B-localized genes and repeat contents. We detected unique sequences occurring exclusively on Bs and discovered various evolutionary patterns of genomic rearrangements associated to Bs. In situ hybridization and quantitative polymerase chain reactions further validated our genomic approach confirming detection of sequences on Bs. The functional annotation of B sequences showed that the B chromosome comprises regions of gene fragments, novel genes, and intact genes, which encode a diverse set of functions related to important biological processes such as metabolism, morphogenesis, reproduction, transposition, recombination, cell cycle and chromosomes functions which might be important for their evolutionary success. This study reveals the genomic structure, composition and function of Bs, which provide new insights for theories of B chromosome evolution. The selfish behavior of Bs seems to be favored by gained genes/sequences.


Efficacy of two mandibular advancement appliances in the management of snoring and mild-moderate sleep apnea: a cross-over randomized study.
Mandibular advancement appliances (MAA) are a recognized alternative treatment to continuous positive airway pressure (CPAP) for mild-moderate obstructive sleep apnea syndrome (OSAS). The aim of this study is to assess the efficacy of and subject satisfaction with two MAA in the management of OSAS. Five women and 11 men (47.9+/-1.6 years), previously untreated with CPAP, were recruited from a sleep disorders clinic following a polysomnographic diagnosis of mild-moderate OSAS with Respiratory Disturbance Index (RDI) of 9.4+/-1.1. A randomized single blind cross-over study was completed with both Klearway and Silencer (three months for each study arm). Subjects completed standardized questionnaires on sleep quality, sleepiness and functional outcomes (Functional Outcome Sleep Questionnaire: FOSQ). MAA satisfaction (e.g., comfort) and efficacy (e.g., reduction of respiratory noises, headache) were assessed by subjects and sleep partner. The two MAA (Silencer 4.7+/-0.9 and Klearway 6.5+/-1.3) significantly reduced the RDI compared to the baseline night (10.0+/-1.2, respectively p<0.001 and p<0.01). The RDI was slightly lower with the Silencer (p0.05) but subjects' preference for comfort was in favor of the Klearway (Klearway 7.0+/-0.4 vs Silencer 5.8+/-0.4, p=0.04). The Epworth score, FOSQ, respiratory noise and morning headache were also improved following use of both appliances (p0.05 to 0.001). Although both MAA decreased RDI and subjective daytime sleepiness in a similar manner, the choice between various types of MAA needs to be taken into account when considering the benefit of RDI reduction over the benefit of subject compliance. The long term benefit of increased RDI reduction vs. a better subject compliance needs to be assessed in prospective studies.


Macrophage migration inhibitory factor induces epithelial to mesenchymal transition, enhances tumor aggressiveness and predicts clinical outcome in resected pancreatic ductal adenocarcinoma.
MIF is a proinflammatory cytokine and is implicated in cancer. A higher MIF level is found in many human cancer and cancer-prone inflammatory diseases, including chronic pancreatitis and pancreatic cancer. We tested the hypothesis that MIF contributes to pancreatic cancer aggressiveness and predicts disease outcome in resected cases. Consistent with our hypothesis we found that an elevated MIF mRNA expression in tumors was significantly associated with poor outcome in resected cases. Multivariate Cox-regression analysis further showed that MIF is independently associated with patients' survival (HR = 2.26, 95% CI = 1.17-4.37, p = 0.015). Mechanistic analyses revealed that MIF overexpression decreased E-cadherin and increased vimentin mRNA and protein levels in pancreatic cancer cell lines, consistent with the features of epithelial-to-mesenchymal transition (EMT). Furthermore, MIF-overexpression significantly increased ZEB1/2 and decreased miR-200b expression, while shRNA-mediated inhibition of MIF increased E-cadherin and miR-200b expression, and reduced the expression of ZEB1/2 in Panc1 cells. Re-expression of miR-200b in MIF overexpressing cells restored the epithelial characteristics, as indicated by an increase in E-cadherin and decrease in ZEB1/2 and vimentin expression. A reduced sensitivity to the chemotherapeutic drug, gemcitabine, occurred in MIF-overexpressing cells. Indicative of an increased malignant potential, MIF over-expressing cells showed significant increase in their invasion ability in vitro, and tumor growth and metastasis in an orthotopic xenograft mouse model. These results support a role of MIF in disease aggressiveness, indicating its potential usefulness as a candidate target for designing improved treatment in pancreatic cancer.


Evaluating the clinical significance of drug interactions: a systematic approach.
Remembering the myriad of psychotropic drug interactions is extremely difficult. Nevertheless, by applying a systematic approach, the clinician can often predict the occurrence and time course of such interactions. Several factors must be considered when assessing the potential consequences. Drug-related factors that increase the risk for clinically significant interactions include a low therapeutic index or narrow therapeutic window, a multiplicity of pharmacological actions, and inhibition or inducement of cytochrome P450 enzymes. Next, patient-related factors that can increase the risk for significant drug interactions should be considered. These include genetically based variations in drug-metabolizing capacity, as well as advanced age, underlying medical illness, and comorbid substance abuse. Finally, the literature should be carefully reviewed to as-certain the potential clinical relevance of available data. If a clinically significant drug interaction appears likely to occur, the patient's clinical status should be followed closely; therapeutic drug monitoring should be used if applicable and dosage adjustments made accordingly. Rational polypharmacy requires an understanding of the pharmacological principles governing drug interactions and a knowledge of the factors that increase the likelihood of clinically significant variations in drug action. This will allow the clinician to maximize beneficial effects while minimizing the risk of adverse events.


Canopy and leaf composition drive patterns of nutrient release from pruning residues in a coffee agroforest.
In a coffee agroforest, the crop is cultivated under the shade of fruit-bearing and nitrogen (N)-fixing trees. These trees are periodically pruned to promote flowering and fruiting as well as to make nutrients stored in tree biomass available to plants. We investigated the effect of canopy composition and substrate quality on decomposition rates and patterns of nutrient release from pruning residues in a coffee agroforest located in Costa Rica's Central Valley. Initial phosphorus (P) release was enhanced under a canopy composed solely of N-fixing, Erythrina poeppigiana compared to a mixed canopy of Erythrina and Musa acuminata (banana). Both initial and final N release were similar under the two canopy types. However, after five months of decomposition, a higher proportion of initial N had been released under the single canopy. Although patterns of decomposition and nutrient release were not predicted by initial substrate quality, mass loss in leaf mixtures rates were well predicted by mean mass loss of their component species. This study identifies specific pruning regimes that may regulate N and P release during crucial growth periods, and it suggests that strategic pruning can enhance nutrient availability. For example, during the onset of rapid fruit growth, a two-species mixture may release more P than a three-species mixture. However, by the time of the harvest, the two- and three-species mixtures have released roughly the same amount of N and P. These nutrients do not always follow the same pattern, as N release can be maximized in single-species substrates, while P release is often facilitated in species mixtures. Our study indicates the importance of management practices in mediating patterns of nutrient release. Future research should investigate how canopy composition and farm management can also mediate on-farm nutrient losses.


Studies on sterol/steroid biosynthesis from 3:4 dihydroxy phenyl alanine.
Homogenated tissue of gonads of Rana tigrina were incubated in Young's Ringer solution containing 5 mg of 3:4 dihydroxy phenylalanine. An increase in the cholesterol and neutral-17-ketosteroids was found when compared with the control. Perfusion of ovary with DOPA gave similar results. However, there was a sharp drop (greater than 50%) in the cholesterol contents of the kidney and suprarenal under similar conditions. A postulated scheme of steroid biosynthesis from DOPA is given.


Ursolic acid induces apoptosis and anoikis in colorectal carcinoma RKO cells.
Ursolic acid (UA) is an anti-cancer herbal compound. In the present study, we observed the effects of UA on anchorage-dependent and -independent growth of human colorectal cancer (CRC) RKO cells. RKO cells were cultured in conventional and detached condition and treated with UA. Cell viability was evaluated by CCK-8 assay. Cell cycle was analyzed by flow cytometry. Apoptosis was identified by Hoechst 33258 staining and flow cytometry analysis. Activities of caspases were measured by commercial kits. Reactive oxygen species (ROS) was recognized by DCFH-DA fluorescent staining. Anoikis was identified by EthD-1 fluorescent staining and flow cytometry analysis. Expression and phosphorylation of proteins were analyzed by western blot. UA inhibited RKO cell viability in both a dose- and time-dependent manner. UA arrested the cell cycle at the G0/G1 phase, and induced caspase-dependent apoptosis. UA inhibited Bcl-2 expression and increased Bax expression. In addition, UA up-regulated the level of ROS that contributed to UA activated caspase-3, - 8 and - 9, and induced apoptosis. Furthermore, UA inhibited cell growth in a detached condition and induced anoikis in RKO cells that was accompanied by dampened phosphorylation of FAK, PI3K and AKT. UA also inhibited epithelial-mesenchymal transition (EMT) as indicated by the down-regulation of N-Cad expression and up-regulation of E-Cad expression. UA induced caspase-dependent apoptosis, and FAK/PI3K/AKT singling and EMT related anoikis in RKO cells. UA was an effective anti-cancer compound against both anchorage-dependent and -independent growth of RKO cells.


Pathogenesis of encephalitis induced in newborn mice by virulent and avirulent strains of Sindbis virus.
Strains of Sindbis virus differ in their virulence for mice of different ages; this variation is related in large part to variations in the amino acid compositions of E1 and E2, the surface glycoproteins. The comparative pathogenesis of Sindbis virus strains which are virulent or avirulent for newborn mice has not been previously examined. We have studied the diseases caused by a virulent wild-type strain, AR339, and two less virulent laboratory strains, Toto1101 and HRSP (HR small plaque). After peripheral inoculation of 1,000 PFU, AR339 causes 100% mortality within 5 days (50% lethal dose [LD50] = 3 PFU) while Toto1101 causes 70% mortality (LD50 = 10(2.4) PFU) and HRSP causes 50 to 60% mortality (LD50 = 10(5.1) PFU) with most deaths occurring 7 to 11 days after infection. However, after intracerebral inoculation of 1,000 PFU, Toto1101 is virulent (100% mortality within 5 days; LD50 = 4 PFU) while HRSP is not (75% mortality; LD50 = 10(4.2) PFU). After intracerebral inoculation, all three strains initiate new virus formation within 4 h, but HRSP reaches a plateau of 10(6) PFU/g of brain while Toto1101 and AR339 replicate to a level of 10(8) to 10(9) PFU/g of brain within 24 h. Interferon induction parallels virus growth. Mice infected with HRSP develop persistent central nervous system infection (10(6) PFU/g of brain) until the initiation of a virus-specific immune response 7 to 8 days after infection when virus clearance begins. The distribution of virus in the brains of mice was similar, but the virus was more abundant in the case of AR339. HRSP continued to spread until day 9. Clearance from the brain was complete by day 17. We conclude that the decreased virulence of HRSP is due to an intrinsic decreased ability of this strain of Sindbis virus to grow in neural cells of the mouse. We also conclude that CD-1 mice do not respond to the antigens of Sindbis virus until approximately 1 week of age. This lack of response does not lead to tolerance and persistent infection but rather to late virus clearance whenever the immune response is initiated.


[The effect of argon-containing hypoxic gas environment on development of Japanese quail embryos].
In case of acute hypoxic hypoxia argon in the gas environment is favorable to survivability of laboratory animals (rats). Our purpose was to evaluate effects of an argon-containing hypoxic gas environment on embryogenesis of the Japanese quail (Cortunix cortunix japonica). Eggs of the Japanese quail were incubated in a pressurized chamber with a hypoxic gas environment differing in concentration of nitrogen and argon. In hypoxic (10% O2) oxygen-nitrogen and oxygen-nitrogen-argon atmospheres total mortality of embryos was observed no later than on day 7 of development. These gas mixtures also had a teratogenic effect. However, about 55% of argon in atmosphere prevented completely microcephalia and reduced in half incidence of other pathologies.


Theoretical effects of fluid infusions during cardiopulmonary resuscitation as demonstrated in a computer model of the circulation.
Recent studies have shown the potential adverse effects of venous volume loading on blood flow during closed chest cardiopulmonary resuscitation (CPR). To examine the effect of arterial and venous infusions, we employed a published computer simulation of the circulation during CPR. This model uses computer simulated electrical networks to model the heart and great vessels. CPR was modeled with compressions at a rate of 80/min and a force of 80 mmHg. Fluid infusions, simulated as current pulses into the abdominal aorta and superior vena cava, were given to measure their effect on myocardial and cranial blood flow. With 600 ml/min infusions into the abdominal aorta, there was a 12% peak increase in myocardial flow and a 3.8% peak increase in cranial flow. Every 100 ml/min increase in infusion from 0 to 900 ml/min produced a 1.4 ml/min linear increase in myocardial flow and a 4.2 ml/min linear increase in cranial flow. In agreement with previous CPR model studies, simulated vasoconstriction of abdominal and lower extremity vessels resulted in increased myocardial and cranial flows. As resistance of these vessels was increased, abdominal aortic infusions resulted in greater flow augmentations. In contrast to arterial results, infusions at 600 ml/min into the vena cava resulted in a 2.2% decrease in myocardial flow and a 0.62% decrease in cranial flow. Rise and fall times for initiation and cessation of flow augmentations were equal to four compression cycles. We conclude that these findings demonstrate the theoretical benefits of rapid arterial infusions during CPR with increases in myocardial and cranial blood flow. This method may provide an early temporary adjunct to myocardial perfusion during CPR.


Whole genome analysis from microdissected tissue revealed adult supratentorial grade II-III gliomas are divided into clinically relevant subgroups by genetic profile.
Gliomas are classified into pathologically defined subgroups. However, the tumors in the same entity could show varied 'clinical courses. Recently, various studies revealed that genetic analyses of gliomas can provide clinically relevant information. To investigate the correlation between genetic characterization and clinical information of adult supratentorial grade II-III gliomas to establish a genetic classification of these tumors. We analyzed the genetic profile of tumor samples microdissected from formalin-fixed archival tissue sections based on the defined selection criteria of tumor region using a comparative genomic hybridization method. We collected genetic and clinical data from 140 adult supratentorial gliomas of World Health Organization grade II-III and classified the tumors analyzed into subgroups according to chromosomal copy number aberrations. The relationship between histologic or genetic subclassification and clinical features was analyzed. The tumors were classified into subgroups based on the genetic profiles. Tumors with +7q and those with -1p/19q showed long progression-free survival, and loss of 10q in association with gain of 7p (+7/-10q) appeared to predict poor outcome. Most tumors with histologic diagnosis of glioblastoma showed +7/-10q; however, World Health Organization grade II-III gliomas contained a variety of genetic subgroups that correlated well with the clinical course, and this might explain why these tumors do not respond uniformly to therapies. Our pilot study suggests that care must be taken in clinical studies of these tumors because, unlike glioblastoma, grade II-III tumors may contain genetically distinct subgroups with different sensitivity to the therapies.


Novel human immunodeficiency virus type 1 protease mutations potentially involved in resistance to protease inhibitors.
Plasma-derived sequences of human immunodeficiency virus type 1 (HIV-1) protease from 1,162 patients (457 drug-naive patients and 705 patients receiving protease inhibitor [PI]-containing antiretroviral regimens) led to the identification and characterization of 17 novel protease mutations potentially associated with resistance to PIs. Fourteen mutations were positively associated with PIs and significantly correlated in pairs and/or clusters with known PI resistance mutations, suggesting their contribution to PI resistance. In particular, E34Q, K43T, and K55R, which were associated with lopinavir treatment, correlated with mutations associated with lopinavir resistance (E34Q with either L33F or F53L, or K43T with I54A) or clustered with multi-PI resistance mutations (K43T with V82A and I54V or V82A, V32I, and I47V, or K55R with V82A, I54V, and M46I). On the other hand, C95F, which was associated with treatment with saquinavir and indinavir, was highly expressed in clusters with either L90M and I93L or V82A and G48V. K45R and K20T, which were associated with nelfinavir treatment, were specifically associated with D30N and N88D and with L90M, respectively. Structural analysis showed that several correlated positions were within 8 A of each other, confirming the role of the local environment for interactions among mutations. We also identified three protease mutations (T12A, L63Q, and H69N) whose frequencies significantly decreased in PI-treated patients compared with that in drug-naive patients. They never showed positive correlations with PI resistance mutations; if anything, H69N showed a negative correlation with the compensatory mutations M36I and L10I. These mutations may prevent the appearance of PI resistance mutations, thus increasing the genetic barrier to PI resistance. Overall, our study contributes to a better definition of protease mutational patterns that regulate PI resistance and strongly suggests that other (novel) mutations beyond those currently known to confer resistance should be taken into account to better predict resistance to antiretroviral drugs.