Combined Analysis of Early CD4+ T Cell Counts and CMV Serostatus May Improve CMV Risk Assessment after Allogeneic Hematopoietic Cell Transplantation.
The incidence and severity of viral complications after cellular therapy are highly variable. Recent publications describe relevant interactions between the human Cytomegalovirus (CMV) and host immunity in recipients of allogeneic hematopoietic cell transplantation (HCT). Although immune monitoring is routinely performed in HCT patients, validated cut-off levels correlating with transplant outcomes such as survival or CMV reactivation are mostly limited to day +100, which is later than the median time for CMV reactivation in the absence of medical prophylaxis. To address this gap in early risk assessment, we applied an unsupervised machine learning technique based on clustering of day +30 CD4+ helper T cell count data, and identified relevant cut-off levels within the diverse spectrum of early CD4+ reconstitution. These clusters were stratified for CMV recipient serostatus to identify early risk groups that predict clinical HCT outcome. Indeed, the new risk groups predicted subsequent clinical events such as NRM, OS, and high CMV peak titers better than the most established predictor, i.e., the positive CMV recipient serostatus (R+). More specifically, patients from the R+/low CD4+ subgroup strongly associated with high CMV peak titers and increased 3-year NRM (subdistribution hazard ratio (SHR) 10.1, 95% CI 1.38-73.8, p = 0.023), while patients from the R-/very high CD4+ subgroup showed comparable NRM risks (SHR 9.57, 95% CI 1.12-81.9, p = 0.039) without such an association. In short, our study established novel cut-off levels for early CD4+ T cells via unsupervised learning and supports the integration of host cellular immunity into clinical risk-assessment after HCT in the context of CMV reactivation.


Evaluation and molecular docking study of two flavonoids from Oroxylum indicum (L.) Kurz and their semi-synthetic derivatives as histone deacetylase inhibitors.
Chrysin (5,7-dihydroxyflavone, 6) and galangin 3-methyl ether (5,7-dihydroxy-3-methoxy flavone, 7) were obtained from the leaves of Oroxylum indicum (L.) Kurz in 4% and 6% yields, respectively. Both compounds could act as pan-histone deacetylase (HDAC) inhibitors. Structural modification of these lead compounds provided thirty-eight derivatives which were further tested as HDAC inhibitors. Compounds 6b, 6c, and 6q were the most potent derivatives with the IC50 values of 97.29 ± 0.63 μM, 91.71 ± 0.27 μM, and 96.87 ± 0.45 µM, respectively. Molecular docking study indicated the selectivity of these three compounds toward HDAC8 and the test against HDAC8 showed IC50 values in the same micromolar range. All three compounds were further evaluated for the anti-proliferative activity against HeLa and A549 cell lines. Compound 6q exhibited the best activity against HeLa cell line with the IC50 value of 13.91 ± 0.34 μM. Moreover, 6q was able to increase the acetylation level of histone H3. These promising HDAC inhibitors deserve investigation as chemotherapeutic agents for treating cancer.


Tailored release of triiodothyronine and retinoic acid from a spatio-temporally fabricated nanofiber composite instigating neuronal differentiation.
Regeneration of the central and peripheral nervous system is challenging since the functional restoration of injured nerves is an incredible task. The fabrication of an ideal nerve guide that fulfills the requirement to regenerate nerve tissue is a herculean challenge requiring a combination of both biochemical and topographical cues. The present study explores the combinatorial effect of aligned nanofibers and the regulated delivery of triiodothyronine and retinoic acid on nerve regeneration. A sequential release mechanism is adopted in fabricating the nanofiber scaffold, with triiodothyronine incorporated into the nanofiber shell ensuring its prior release, followed by retinoic acid (entrapped within zein nanoparticles) from the core. The composite nanofibers thus fabricated possess excellent mechanical, physical and thermal properties and good topographical morphology and were highly biocompatible. The nanofibers were scrutinized for their efficacy in stimulating differentiation to a neuronal phenotype. The elongation factor (E-factor) of the neural cells had doubled in the bioactive incorporated composite compared to other scaffolds, as observed on phalloidin staining of their cytoskeleton, which endorsed enhanced neural differentiation on the fabricated nanofiber scaffold. There was a significant increase in the expression of neural-lineage specific markers on investigation of mRNA by real time PCR, showing a 10 fold increase in the gene expression of β-III-tubulin, a 5.5 fold increase for microtubule associated protein 2 gene and 3.5 fold for neurofilament M gene in the cells cultured over bioactive incorporated aligned nanofiber composites. Similarly protein expression was analyzed by immunofluorescence and flow cytometry studies, which showed an increase in the expression of β-III-tubulin in the composite nanofiber. This corroborates that neuronal differentiation is enhanced by the aligned nanotopography and spatio-temporal delivery of triiodothyronine and retinoic acid, opening avenues for nerve regenerative graft fabrication.


Comparison of peak cardiopulmonary performance parameters on a robotics-assisted tilt table, a cycle and a treadmill.
Robotics-assisted tilt table (RATT) technology provides body support, cyclical stepping movement and physiological loading. This technology can potentially be used to facilitate the estimation of peak cardiopulmonary performance parameters in patients who have neurological or other problems that may preclude testing on a treadmill or cycle ergometer. The aim of the study was to compare the magnitude of peak cardiopulmonary performance parameters including peak oxygen uptake (VO2peak) and peak heart rate (HRpeak) obtained from a robotics-assisted tilt table (RATT), a cycle ergometer and a treadmill. The strength of correlations between the three devices, test-retest reliability and repeatability were also assessed. Eighteen healthy subjects performed six maximal exercise tests, with two tests on each of the three exercise modalities. Data from the second tests were used for the comparative and correlation analyses. For nine subjects, test-retest reliability and repeatability of VO2peak and HRpeak were assessed. Absolute VO2peak from the RATT, the cycle ergometer and the treadmill was (mean (SD)) 2.2 (0.56), 2.8 (0.80) and 3.2 (0.87) L/min, respectively (p < 0.001). HRpeak from the RATT, the cycle ergometer and the treadmill was 168 (9.5), 179 (7.9) and 184 (6.9) beats/min, respectively (p < 0.001). VO2peak and HRpeak from the RATT vs the cycle ergometer and the RATT vs the treadmill showed strong correlations. Test-retest reliability and repeatability were high for VO2peak and HRpeak for all devices. The results demonstrate that the RATT is a valid and reliable device for exercise testing. There is potential for the RATT to be used in severely impaired subjects who cannot use the standard modalities.


The reliability and validity of a short FFQ among Australian Aboriginal and Torres Strait Islander and non-Indigenous rural children.
To determine the reproducibility and validity of a short FFQ (SFFQ) for Australian rural children aged 10 to 12 years, particularly Aboriginal and Torres Strait Islander children. In this cross-sectional study participants completed the SFFQ on two occasions and three 24 h recalls. Concurrent validity was established by comparing results of the first SFFQ against food recalls; reproducibility was established by comparing the two SFFQ. The north coast of New South Wales in the Australian summer of late 2005. Two hundred and forty-one children (ninety-two Aboriginal and Torres Strait Islander children and 100 boys) completed two SFFQ and were included in the reproducibility study; of these, 205 participants with a mean age of 10·8 (sd 0·7) years took part in the validity study. The SFFQ showed moderate to good reproducibility among all children with kappa coefficients for repeated measures between 0·41 and 0·80. Eighteen of twenty-three questions demonstrated good validity against the mean of the 24 h recalls, with statistically significant increasing trends (P ≤ 0·05) for mean daily weight and/or frequency as survey response categories increased. A similar number of short questions showed good validity for Aboriginal and Torres Strait Islander children as for their non-Indigenous counterparts. Many short questions in this SFFQ are able to discriminate between different categories of food intake and provide information on relative intake within the given population. They can be used to monitor and/or evaluate population-wide health programmes, including those with rural Aboriginal and Torres Strait Islander children.


Effects of garlic supplementation on liver enzymes: A systematic review and meta-analysis of randomized controlled trials.
Current evidence on the beneficial effects of garlic on liver enzymes is contradictory. Therefore, the aim of this systematic review and meta-analysis is to evaluate the effect of garlic supplementation on human liver enzymes, such as Alanine Transaminase (ALT/SGPT) and Aspartate Transaminase (AST/SGOT). To collect the required data, PubMed, Scopus, ISI Web of Science, and Google scholar databases were systematically searched from inception to June 2019. A meta-analysis was conducted using the random-effects model to evaluate the effects of garlic supplementation on ALT and AST levels. The Cochran's Q-test and inconsistency index were also used to evaluate heterogeneity among the studies. Among a total of 15,514 identified articles, six studies (containing 301 participants) met the inclusion criteria. Results of the meta-analysis showed that garlic supplementation significantly decreased AST level (Hedges' g = -0.36, 95% confidence interval [CI]: -0.72, -0.004, p = .047); whereas, it had no significant effect on ALT level (Hedges' g = -0.22, 95% CI: -0.64, 0.20, p = .310). Results showed that garlic supplementation reduced AST levels significantly; however, had no significant effect on ALT levels. Further studies are still needed to confirm the results.


Daucus carota Pentane/Diethyl Ether Fraction Inhibits Motility and Reduces Invasion of Cancer Cells.
Daucus carota (DC) is a herb used in folklore medicine in Lebanon to treat numerous diseases including cancer. Recent studies in our laboratory on DC oil and its fractions revealed potent anticancer activities in vitro and in vivo. The present study aims to investigate the effect of the most potent DC fraction, pentane/diethyl ether (50:50), on lung, skin, breast and glioblastoma cancer cell motility and invasion. Upon treatment, a pronounced decrease in cancer cell motility was observed in the 4 cell lines. The treatment also led to a decrease in cancer cell invasion and an increased cell adhesion. Additionally, the DC fraction caused a decrease in the activation of the ρ-GTPases Rac and CDC42, a finding that may partially explain the treatment-induced decrease in cell motility. The current study demonstrates a crucial effect of the DC pentane/diethyl ether fraction on cancer cell motility and metastasis, making it a potential candidate for cancer therapy specifically targeting cancer motility and metastasis.


13-cis-retinoic acid and interferon alpha-2a: effective combination therapy for advanced squamous cell carcinoma of the skin.
Retinoids (vitamin A derivatives) and interferon-alpha (IFN-alpha) are potent regulators of malignant cell differentiation and proliferation, and both have immunomodulatory and antiangiogenesis activity. A large body of preclinical and clinical data supports the use of combination therapy with 13-cis-retinoic acid (13-cRA) and IFN-alpha in patients with squamous cell carcinoma of the skin. This carcinoma is an extremely common and frequently severely disfiguring cancer, for which about 10% of patients remain uncured following standard local therapy. Our purpose was to test whether a 20% or greater complete response rate could be achieved using a combination of these two agents in patients with advanced squamous cell carcinoma of the skin in whom local therapy had failed or was unfeasible or who had regional and/or distant metastases. Thirty-two patients with heavily pretreated, advanced inoperable cutaneous squamous cell carcinoma of the skin were given a combination of oral 13-cRA (1 mg/kg per day) and subcutaneous recombinant human IFN alpha-2a (3 million units per day) for at least 2 months, unless disease progressed earlier, in a phase II trial. Nineteen (68%) of the 28 assessable patients responded, seven (25%) of whom had complete responses. Response rates were 93% (13 of 14) in patients with advanced local disease (six complete responses), 67% (four of six) in patients with regional disease (no complete responses), and 25% (two of eight) in patients with distant metastases (one complete response). The relationship between decreased response rate and increased extent of disease was highly statistically significant (P less than .005, chi-square test). The median response duration was greater than 5 months. No life-threatening toxic effects occurred in assessable patients treated with this combination, although dose reductions were required in 18 patients. The major limiting side effect in this elderly patient population (median age, 67 years) was cumulative fatigue. These results indicate that combined systemic therapy with 13-cRA and IFN alpha-2a is highly effective in patients with advanced squamous cell carcinoma of the skin.


Essential oils as biopreservatives: different methods for the technological application in lettuce leaves.
Investigations were carried out to assess the efficiency of 3 essential oils, clove, tea tree, and rosemary, as natural preservatives during the postharvest of lettuce leaves. The effect of different concentration (1 and 0.5 MIC) of plant essential oils applied in 3 forms (spray, immersion, and capsules) was studied on lettuce leaves. The evolution of different microbial populations was evaluated during refrigerated storage. The application forms of the biopreservatives were shown to be an important factor in determining the effectiveness of the essential oils. Clove and tea tree essential oils at 1 MIC and applied embedded in lactose capsules presented a significant inhibition on mesophilic, psicrotrophic, and coliforms populations, while rosemary in none of the 3 technological applications forms exerted inhibitory effect on all microbial populations evaluated. Essential oils (at 0.5 MIC) applied by spray, immersion, and embedded in lactose capsules exerted lower inhibitory effects, with respect to 1 MIC, on the different microbial populations present on lettuce leaves. At the end of the storage (7 d), lettuce samples treated with tea tree, clove, and rosemary (at 1 and 0.5 MIC) by spray were the only organoleptically acceptable. It is concluded that clove and tea tree essential oils can control different microbial population present in lettuce. Practical Application: The exploration of naturally occurring antimicrobials in food preservation receives increasing attention due to consumer awareness of natural food products. Biopreservatives are useful in extending the shelf life of foods, reducing or eliminating pathogenic bacteria and increasing overall quality of food products. The effectiveness of essential oil application in foods is the result of factor associations such as applications forms, concentration applied, the way of action, storage temperatures. The application methods (spray, immersion, and embedded in lactose capsules) and the concentration of essential oils have been shown to be important factors in determining the effectiveness of these biopreservatives. The oil concentrations required to produce a certain level of inhibition in actual foods could be questionable due to the organoleptic impact. However, these novel natural preservatives in combination with other factors in obstacle technologies are an alternative to control the pathogen growth minimizing undesirable changes in organoleptic characteristics.


Extensive polymorphism in the porcine Toll-like receptor 10 gene.
The great importance of the Toll-like receptors (TLRs) in innate immunity is well established, but one family member--TLR10--remains elusive. TLR10 is expressed in various tissues in several species, but its ligand is not known and its function is still poorly understood. The open reading frame of TLR10 was sequenced in 15 wild boars, representing three populations, and in 15 unrelated domestic pigs of Hampshire, Landrace and Large White origin. Amino acid positions corresponding to detected nonsynonymous single nucleotide polymorphisms (SNPs) were analysed in the crystal structures determined for the human TLR1-TLR2-lipopeptide complex and the human TLR10 Toll/Interleukin 1 receptor (TIR) dimer. SNP occurrence in wild boars and domestic pigs was compared, and haplotypes for the TLR10 gene and the TLR6-1-10 gene cluster were reconstructed. Despite the limited number of animals sequenced in the present study (N = 30), a larger number of SNPs were found in TLR10 than recently reported for TLR1, TLR6 and TLR2. Thirty-three SNPs were detected, of which 20 were nonsynonymous. The relative frequency of nonsynonymous (d(N) ) and synonymous (d(S) ) SNPs between wild boars and domestic pigs was higher in TLR10 than recently reported for TLR1, TLR6 and TLR2. However, the polymorphism reported in the present study seems to leave the function of the TLR10 molecule unaffected. Furthermore, no nonsynonymous SNPs were detected in the part of the gene corresponding to the hinge region of the receptor, probably reflecting rigorously acting functional constraint. The total number of SNPs and the number of nonsynonymous SNPs were significantly lower (P < 0.05) in the wild boars than in the domestic pigs, and fewer TLR10 haplotypes were present in the wild boars. The majority of the TLR6-1-10 haplotypes were specific for either wild boars or domestic pigs, probably reflecting differences in microbial environment and population history.


Mechanical forces induce odontoblastic differentiation of mesenchymal stem cells on three-dimensional biomimetic scaffolds.
The mechanical induction of cell differentiation is well known. However, the effect of mechanical compression on odontoblastic differentiation remains to be elucidated. Thus, we first determined the optimal conditions for the induction of human dental pulp stem cells (hDPSCs) into odontoblastic differentiation in response to mechanical compression of three-dimensional (3D) scaffolds with dentinal tubule-like pores. The odontoblastic differentiation was evaluated by gene expression and confocal laser microscopy. The optimal conditions, which were: cell density, 4.0 × 105 cells/cm2 ; compression magnitude, 19.6 kPa; and loading time, 9 h, significantly increased expression of the odontoblast-specific markers dentine sialophosphoprotein (DSPP) and enamelysin and enhanced the elongation of cellular processes into the pores of the membrane, a typical morphological feature of odontoblasts. In addition, upregulation of bone morphogenetic protein 7 (BMP7) and wingless-type MMTV integration site family member 10a (Wnt10a) was observed. Moreover, the phosphorylation levels of mitogen-activated protein kinases (MAPKs), extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 were also enhanced by mechanical compression, indicating the involvement of the MAPK signalling pathway. It is noteworthy that human mesenchymal stem cells (MSCs) derived from bone marrow and amnion also differentiated into odontoblasts in response to the optimal mechanical compression, demonstrating the importance of the physical structure of the scaffold in odontoblastic differentiation. Thus, odontoblastic differentiation of hDPSCs is promoted by optimal mechanical compression through the MAPK signalling pathway and expression of the BMP7 and Wnt10a genes. The 3D biomimetic scaffolds with dentinal tubule-like pores were critical for the odontoblastic differentiation of MSCs induced by mechanical compression. Copyright © 2014 John Wiley & Sons, Ltd.


Salt and peroxide compared with conventional oral hygiene. I. Clinical results.
The purposes of this 2-year longitudinal study were to: compare the clinical effectiveness of patient applied sodium bicarbonate, hydrogen peroxide, and sodium chloride (S/P) to the use of conventional oral hygiene methods and to investigate the motivational effect of using phase-contrast microscopy in teaching effective oral hygiene. Initially, 972 subjects were screened for signs of periodontitis. From these, 347 with early to moderate periodontitis were selected and each was randomly assigned to one of four home treatment regimens after scaling and root planing. The four treatment regimens included: conventional oral hygiene procedures, conventional oral hygiene procedures plus phase-contrast demonstration of subgingival microbial forms for oral hygiene motivation, S/P oral hygiene, and S/P oral hygiene plus phase-contrast demonstration of subgingival microbial forms for oral hygiene motivation. Plaque, bleeding, gingival inflammation, probing depth, and clinical attachment level were recorded at baseline, 8, 16, and 24 months. Subjects were recalled for reinforcement of oral hygiene and periodontal prophylaxis at various intervals. Data were analyzed based on disease severity, location of index sites and compliance. The results indicated that both conventional oral hygiene procedures and the S/P regimen were effective in reducing clinical signs of disease when combined with professional care. There were no differences between the two regimens in clinical effectiveness and trends favoring microscopic viewing of subgingival plaque for motivational purposes were not statistically significant.


Congenital Adrenal Hyperplasia and Schmid Metaphyseal Chondrodysplasia in a Child.
Congenital adrenal hyperplasia (CAH) is a group of hereditary diseases, which are autosomal recessive. CAH occurs due to defect in one of the cortisol coding genes and often clinically presents itself with signs of androgen overproduction. In this article, we report a case of CAH and Schmid metaphyseal dysplasia. Our literature review indicated that this report is the first attempt on CYP11B1 and Schmid dysplasia in a child. The specific diagnosis of 11-β-hydroxylase deficiency can be determined using high basal levels of deoxycorticosterone and/or 11-deoxycortisol serums.


Tissue transglutaminase_variant 2-transduced mesenchymal stem cells and their chondrogenic potential.
As cartilage is incapable of self-healing upon severe degeneration because of the lack of blood vessels, cartilage tissue engineering is gaining importance in the treatment of cartilage defects. This study was designed to improve cartilage tissue regeneration by expressing tissue transglutaminase variant 2 (TGM2_v2) in mesenchymal stem cells (MSC) derived from bone marrow of rats. For this purpose, rat MSCs transduced with TGM2_v2 were grown and differentiated on three-dimensional polybutylene succinate (PBSu) and poly-l-lactide (PLLA) blend scaffolds. The transduced cells could not only successfully express the short form transglutaminase-2, but also deposited the protein onto the scaffolds. In addition, they could spontaneously produce cartilage-specific proteins without any chondrogenic induction, suggesting that TGM2_v2 expression provided the cells the ability of chondrogenic differentiation. PBSu:PLLA scaffolds loaded with TGM2_v2 expressing MSCs could be used in repair of articular cartilage defects.


Macro-ecology of Gulf of Mexico cold seeps.
Shortly after the discovery of chemosynthetic ecosystems at deep-sea hydrothermal vents, similar ecosystems were found at cold seeps in the Gulf of Mexico. Over the past two decades, these sites have become model systems for understanding the physiology of the symbiont-containing megafauna and the ecology of seep communities worldwide. Symbiont-containing bi-valves and siboglinid polychaetes dominate the communities, including five bathymodiolin mussel species and six vestimentiferan (siboglinid polychaete) species in the Gulf of Mexico. The mussels include the first described examples of methanotrophic symbiosis and dual methanotrophic/thiotrophic symbiosis. Studies with the vestimentiferans have demonstrated their potential for extreme longevity and their ability to use posterior structures for subsurface exchange of dissolved metabolites. Ecological investigations have demonstrated that the vestimentiferans function as ecosystem engineers and identified a community succession sequence from a specialized high-biomass endemic community to a low-biomass community of background fauna over the life of a hydrocarbon seep site.


Biochemical activities of T-antigen proteins encoded by simian virus 40 A gene deletion mutants.
We have analyzed T antigens produced by a set of simian virus 40 (SV40) A gene deletion mutants for ATPase activity and for binding to the SV40 origin of DNA replication. Virus stocks of nonviable SV40 A gene deletion mutants were established in SV40-transformed monkey COS cells. Mutant T antigens were produced in mutant virus-infected CV1 cells. The structures of the mutant T antigens were characterized by immunoprecipitation with monoclonal antibodies directed against distinct regions of the T-antigen molecule. T antigens in crude extracts prepared from cells infected with 10 different mutants were immobilized on polyacrylamide beads with monoclonal antibodies, quantified by Coomassie blue staining, and then assayed directly for T antigen-specific ATPase activity and for binding to the SV40 origin of DNA replication. Our results indicate that the T antigen coding sequences required for origin binding map between 0.54 and 0.35 map units on the SV40 genome. In contrast, sequences closer to the C terminus of T antigen (between 0.24 and 0.20 map units) are required for ATPase activity. The presence of the ATPase activity correlated closely with the ability of the mutant viruses to replicate and to transform nonpermissive cells. The origin binding activity was retained, however, by three mutants that lacked these two functions, indicating that this activity is not sufficient to support either cellular transformation or viral replication. Neither the ATPase activity nor the origin binding activity correlated with the ability of the mutant DNA to activate silent rRNA genes or host cell DNA synthesis.


Cerebral microdialysis and intracranial pressure monitoring in patients with idiopathic normal-pressure hydrocephalus: association with clinical response to extended lumbar drainage and shunt surgery.
This study was performed in patients with idiopathic normal-pressure hydrocephalus (iNPH) to monitor cerebral metabolism with microdialysis (MD) and intracranial pressure (ICP) readings, and relate to the clinical responses to extended lumbar drainage (ELD) and shunt surgery. The baseline levels of MD metabolites and ICP were monitored overnight in 40 consecutive patients with iNPH. In a subset of 28 patients, monitoring was continued during 3 days of ELD. Thirty-one patients received a ventriculoperitoneal shunt. The clinical severity of iNPH was determined before and then 3 and 6-12 months after shunt surgery. Altered levels of MD markers (lactate, pyruvate, lactate/pyruvate ratio, glutamate, and/or glycerol) were seen in all patients at baseline; these improved during ELD. Despite normal static ICP (mean ICP), the pulsatile ICP (the ICP wave amplitude) was increased in 24 patients (60%). Only the level of the ICP wave amplitude differentiated the ELD and/or shunt responders from nonresponders. The MD monitoring indicated low-grade cerebral ischemia in patients with iNPH; during ELD, cerebral metabolism improved. The pulsatile ICP (the ICP wave amplitude) was the only variable differentiating the clinical responders from the nonresponders. The authors suggest that the pulsatile ICP reflects the intracranial compliance and that CSF diversion improves the biophysical milieu of the nerve cells, which subsequently may improve their biochemical milieu.


Global Spread of Norovirus GII.17 Kawasaki 308, 2014-2016.
Analysis of complete capsid sequences of the emerging norovirus GII.17 Kawasaki 308 from 13 countries demonstrated that they originated from a single haplotype since the initial emergence in China in late 2014. Global spread of a sublineage SL2 was identified. A new sublineage SL3 emerged in China in 2016.


Acute Intestinal Obstruction Due to Ileocolic Intussusception in an Adult; A Rare Presentation of Inflammatory Myofibroblastic Tumor.
BACKGROUND Intussusception is not very common in adults, and acute intestinal obstruction with intussusception due to inflammatory myofibroblastic tumor (IMT) is extremely rare. IMT is an uncommon lesion and has no single defined cause. It predominantly affects the pediatric age group and commonly involves the lungs. Here we present a case of IMT causing ileocolic intussusception leading to acute intestinal obstruction in an adult. CASE REPORT A 40-year-old female came to the emergency department with severe colicky pain in her abdomen, and reported 6 to 7 episodes of vomiting with bilious contents, along with an inability to pass feces and flatus for 3 days. An x-ray of her abdomen in erect posture revealed multiple air-fluid levels. Because she had a previous history of tuberculosis, a possible tubercular stricture as the cause of her acute obstruction was considered; an exploratory laparotomy was performed showing her bowel loops were dilated with ileocolic intussusception. The lead point of intussusception (a well-defined 4×4×3.5 cm solid mass), was found at 15 cm proximal to the ileocecal junction. A right hemicolectomy with ileo-transverse anastomosis was performed. The histopathological examination confirmed the presence of IMT. CONCLUSIONS IMT causing ileocolic intussusception with acute intestinal obstruction is an extremely rare presentation of an uncommon entity in adults. High index of suspicion, and appropriate investigations (x-ray abdomen, ultrasound, computed tomography, and colonoscopy) depending on presentation and clinical condition of the patient can result in prompt diagnosis and early management.


Reducing the global burden of hazardous alcohol use: a comparative cost-effectiveness analysis.
Intervention strategies are available for reducing the high global burden of hazardous alcohol use as a risk factor for disease, but little is known about their potential costs and effects at a population level. This study set out to estimate these costs and effects. Analyses were carried out for 12 epidemiological World Health Organization subregions of the world. A population model was used to estimate the impact of evidence-based personal and nonpersonal interventions--including brief physician advice, taxation, roadside random breath testing, restricted sales access and advertising bans. Costs were measured in international dollars (I$); population-level intervention effects were gauged in terms of disability-adjusted life years (DALYs) averted. Average and incremental cost-effectiveness ratios (CERs) were computed. The most costly interventions to implement are brief advice in primary care and roadside breath testing of drivers. In populations with a high prevalence of heavy drinkers (more than 5%, such as Europe and North America), the most effective and cost-effective intervention was taxation (more than 500 DALYs averted per 1 million population; CER < I$500 per DALY averted). In populations with a lower prevalence of heavy drinking, however, taxation is estimated to be less cost effective overall than other, more targeted strategies, such as brief physician advice, roadside breath testing and advertising bans. The most efficient public health response to the burden of alcohol use depends on the prevalence of hazardous alcohol use, which is related to overall per capita consumption. Population-wide measures, such as taxation, are expected to represent the most cost-effective response in populations with moderate or high levels of drinking, whereas more targeted strategies are indicated in populations with lower rates of hazardous alcohol use.


Functional magnetic resonance imaging of delay and trace eyeblink conditioning in the primary visual cortex of the rabbit.
The primary sensory cortices have been shown in recent years to undergo experience- and learning-related plasticity under a variety of experimental circumstances. In this study, we used functional magnetic resonance imaging (fMRI) in parallel with both delay and trace eyeblink conditioning to image the learning-related functional activation within the primary visual cortex (V1) of awake, behaving rabbits. We expected that the differing level of forebrain dependence between these two conditioning paradigms should produce a differential blood oxygenation level-dependent (BOLD) functional response in V1. Our results showed a significant expansion of activated volume within V1, particularly early in learning, after training with the more cognitively demanding trace paradigm. In contrast, the simpler delay paradigm produced an increase in the magnitude of the BOLD response in activated voxels, but no significant change in activated volume. No accompanying learning-related changes were observed in the primary somatosensory cortex, which mediates the unconditioned stimulus. These results suggest that the recruitment of additional neurons within V1 is necessary to support the more demanding memory imposed by the trace interval. To our knowledge, this work is the first functional imaging study to compare directly trace and delay eyeblink conditioning in an animal model.


Evolutionary strategies of viruses, bacteria and archaea in hydrothermal vent ecosystems revealed through metagenomics.
The deep-sea hydrothermal vent habitat hosts a diverse community of archaea and bacteria that withstand extreme fluctuations in environmental conditions. Abundant viruses in these systems, a high proportion of which are lysogenic, must also withstand these environmental extremes. Here, we explore the evolutionary strategies of both microorganisms and viruses in hydrothermal systems through comparative analysis of a cellular and viral metagenome, collected by size fractionation of high temperature fluids from a diffuse flow hydrothermal vent. We detected a high enrichment of mobile elements and proviruses in the cellular fraction relative to microorganisms in other environments. We observed a relatively high abundance of genes related to energy metabolism as well as cofactors and vitamins in the viral fraction compared to the cellular fraction, which suggest encoding of auxiliary metabolic genes on viral genomes. Moreover, the observation of stronger purifying selection in the viral versus cellular gene pool suggests viral strategies that promote prolonged host integration. Our results demonstrate that there is great potential for hydrothermal vent viruses to integrate into hosts, facilitate horizontal gene transfer, and express or transfer genes that manipulate the hosts' functional capabilities.


Brownian dynamics simulations of one-patch inverse patchy particles.
Inverse patchy particles are promising colloids to develop new architectures in ceramic materials based on their self-assembly. Nonetheless, a good understanding of their aggregation is required. Several previous studies have shown that the behavior of ceramic colloids can be well described by the DLVO interaction potential. In the present paper, we develop new coarse-grained Brownian dynamics simulations, where particles are represented by an assembly of beads interacting via DLVO interactions, whose parameters can be directly linked to experimental characterization. First, the validity of the simulations is proved by studying the heteroaggregation of homogeneously charged particles. Then, simulations are applied to one-patch inverse patchy particles to study the effect of the patch size. They show that the smaller the patch, the more elongated the aggregates. Simulations are also performed to understand the role of the Debye screening length in the particular case of large patches and they show that aggregation leads always to compact aggregates.


Doubly labeled water analysis using cavity ring-down spectroscopy.
The doubly labeled water method provides an objective and accurate measure of total energy expenditure in free-living subjects and is considered the gold-standard method for this measurement. Its use, however, is limited by the need to employ isotope ratio mass spectrometry (IRMS) to obtain the high-precision isotopic abundance analyses needed to optimize the dose of expensive (18) O-labeled water. Recently, cavity-ring down spectroscopy (CRDS) instruments have become commercially available and may serve as a less expensive alternative to IRMS. We compared the precision and accuracy of CRDS with those of IRMS for the measurement of total energy expenditure from urine specimens in 14 human subjects. The relative accuracy and precision (SD) for total body water was 0.5 ± 1% and for total energy expenditure was 0.5 ± 6%. The CRDS instrument displayed a memory between successive specimens of 5% for (18) O and 9% for (2) H. The memory necessitated carefully ordering of specimens to reduce isotopic disparity, performance of several injections of each specimen to condition the analyzer, and use of a mathematical memory correction on subsequent injections. These limited the specimen throughput to about one urine specimen per hour. CRDS provided accuracy and precision for isotope abundance measurements of urine that were comparable with those of IRMS. The memory problems were easily recognized by our experienced laboratory staff, but future efforts should be aimed at reducing the memory of the CRDS so that it would be less likely to result in poor reproducibility in laboratories using doubly labeled water for the first time.


[Diabetic pregnancy: outpatient follow-up in a Brazilian University Hospital].
In this study we assessed neonatal complications of diabetic in 50 pregnant women at a University Hospital during 2001-2002: 13 (26%) with type 1 diabetes (DM1), 16 with DM2, and 21 (42%) with gestational DM (GDM). The mean outpatient follow-up was at 16.3+/-8 wk for patients with DM1, 22.9+/-7.5 wk for DM2, and 26.0+/-8.9 wk for GDM. Mean HbA1c, fasting and 2-h post-prandial glycemia on first attendance were respectively: 6.1+/-1,1% (RV: 2.6-6.2%), 132+/-39 mg/dL and 190+/-54 mg/dL. 22 patients were on insulin and 15 were on oral antidiabetic agents (OA) at first evaluation. OA were taken on conception and during the first pregnancy trimester and no malformations were seen in the children. Their metabolic profile was similar to other pregnant women. Caesarean section was needed in 54.5% of deliveries. 56.1% were macrosomic babies, with a mean fetal weight of 3.48+/-0.73 Kg, with no differences according to treatment (insulin vs. OA). We conclude that diabetic pregnant women begin their prenatal care at a later period, often taking OA that are not officially advised to be used during pregnancy and are not in a regular metabolic control. As a result, they have macrosomic infants. Even though we have found no complications related to the OA use during pregnancy, we should not encourage their use until more safety studies are available.


Functional Changes in the Gut Microbiome Contribute to Transforming Growth Factor β-Deficient Colon Cancer.
Colorectal cancer (CRC) is one of the most treatable cancers, with a 5-year survival rate of ~64%, yet over 50,000 deaths occur yearly in the United States. In 15% of cases, deficiency in mismatch repair leads to null mutations in transforming growth factor β (TGF-β) type II receptor, yet genotype alone is not responsible for tumorigenesis. Previous work in mice shows that disruptions in TGF-β signaling combined with Helicobacter hepaticus cause tumorigenesis, indicating a synergistic effect between genotype and microbial environment. Here, we examine functional shifts in the gut microbiome in CRC using integrated -omics approaches to untangle the role of host genotype, inflammation, and microbial ecology. We profile the gut microbiome of 40 mice with/without deficiency in TGF-β signaling from a Smad3 (mothers against decapentaplegic homolog-3) knockout and with/without inoculation with H. hepaticus. Clear functional differences in the microbiome tied to specific bacterial species emerge from four pathways related to human colon cancer: lipopolysaccharide (LPS) production, polyamine synthesis, butyrate metabolism, and oxidative phosphorylation (OXPHOS). Specifically, an increase in Mucispirillum schaedleri drives LPS production, which is associated with an inflammatory response. We observe a commensurate decrease in butyrate production from Lachnospiraceae bacterium A4, which could promote tumor formation. H. hepaticus causes an increase in OXPHOS that may increase DNA-damaging free radicals. Finally, multiple bacterial species increase polyamines that are associated with colon cancer, implicating not just diet but also the microbiome in polyamine levels. These insights into cross talk between the microbiome, host genotype, and inflammation could promote the development of diagnostics and therapies for CRC. IMPORTANCE Most research on the gut microbiome in colon cancer focuses on taxonomic changes at the genus level using 16S rRNA gene sequencing. Here, we develop a new methodology to integrate DNA and RNA data sets to examine functional shifts at the species level that are important to tumor development. We uncover several metabolic pathways in the microbiome that, when perturbed by host genetics and H. hepaticus inoculation, contribute to colon cancer. The work presented here lays a foundation for improved bioinformatics methodologies to closely examine the cross talk between specific organisms and the host, important for the development of diagnostics and pre/probiotic treatment.


Epidural analgesia-anaesthesia in obstetrics.
Epidural analgesia is the most effective and innocuous technique for obstetrics. Pain relief is its main indication but maternal diseases that might be decompensated by labour and delivery are also accepted indications. Low doses of long-acting local anaesthetics alone or in combination with low doses of fentanyl or sufentanil provide good quality analgesia and are safe for mother and fetus. Test doses in parturients lack sufficient specificity and sensitivity for detecting inadvertent intravascular injection, and subarachnoid migration of the catheter is possible at any time during the procedure. Therefore, every injection must be considered as a test dose and only fractionated injections must be made. Epidural block to T10 is needed for labour and to level T4 for Caesarean section. Maintenance of the block with a continuous infusion, or patient-controlled epidural analgesia with a background continuous infusion, provides more stable analgesia than by intermittent injection. Technical difficulties, dural tap, bloody tap, hypotension and insufficient block are most frequent complications of epidural block in obstetrics. Excessive motor block prolongs the second stage of labour and increases the frequency for instrumental delivery and is therefore considered a complication.


Five-year treatment outcomes for teeth with large amalgams and crowns.
For 4735 posterior complex amalgams and crowns placed in adults with continuous dental HMO coverage, all additional treatment received over the subsequent 5 years was determined. The restorations were placed under routine clinical conditions by 74 different dentists among a broad spectrum of insured dental patients. Treatment outcomes were described in terms of a hierarchical classification of additional treatments. At the extremes, a successful outcome was defined as no additional treatment or an additional one- or two-surface restoration on the same tooth, and a catastrophic outcome as extraction or endodontic treatment. Due to clinical protocols, teeth with guarded to poor prognosis prior to treatment are overrepresented in the five-surface amalgam cohort. Successful outcomes characterized 72% of four-surface amalgams, 65% of five-surface amalgams, 84% of gold crowns, and 84% of porcelain crowns. Catastrophic outcomes occurred for 10% of four-surface amalgams, 15% of five-surface amalgams, 8% of gold crowns, and 9% of porcelain crowns.


Application of Machine Learning in Intensive Care Unit (ICU) Settings Using MIMIC Dataset: Systematic Review.
Modern Intensive Care Units (ICUs) provide continuous monitoring of critically ill patients susceptible to many complications affecting morbidity and mortality. ICU settings require a high staff-to-patient ratio and generates a sheer volume of data. For clinicians, the real-time interpretation of data and decision-making is a challenging task. Machine Learning (ML) techniques in ICUs are making headway in the early detection of high-risk events due to increased processing power and freely available datasets such as the Medical Information Mart for Intensive Care (MIMIC). We conducted a systematic literature review to evaluate the effectiveness of applying ML in the ICU settings using the MIMIC dataset. A total of 322 articles were reviewed and a quantitative descriptive analysis was performed on 61 qualified articles that applied ML techniques in ICU settings using MIMIC data. We assembled the qualified articles to provide insights into the areas of application, clinical variables used, and treatment outcomes that can pave the way for further adoption of this promising technology and possible use in routine clinical decision-making. The lessons learned from our review can provide guidance to researchers on application of ML techniques to increase their rate of adoption in healthcare.


The elusive permeability barriers and binding sites for proflavine in Escherichia coli.
Cells of proflavine-sensitive and -resistant Escherichia coli strains were altered in different ways, and the proflavine binding of the changed material was studied. Spheroplasts prepared from sensitive and resistant cells bound similar amounts of proflavine at saturation, whether or not they were osmotically protected by 10% sucrose. Intact cells bound approximately the same amounts of proflavine as spheroplasts. On addition of glucose, osmotically protected resistant but not sensitive spheroplasts released proflavine; unprotected spheroplasts did not release bound proflavine. Thus, osmotically protected membranes are not required for proflavine binding (a passive process) but are required for proflavine release (an active process). The presence of sucrose reduced proflavine binding by resistant cells. Adding glucose to cells in 20% sucrose did not cause a release of residual proflavine, though glucose caused a release of proflavine from cells suspended in 0 or 10% sucrose. On treatment of heated cells or ruptured spheroplasts with nucleases and Pronase, practically all nucleic acids were removed. Proflavine-binding ability of such preparations fell by only 30 to 50%. Washing heated cells with ethanol did not reduce their proflavine-binding ability. There appear to be important binding sites in cells aside from nucleic acids.


Biliary complications of living related pediatric liver transplant patients.
Patients who undergo living related left lateral segment liver transplants have been reported to have a high incidence of biliary complications and some studies suggest that most patients will ultimately need operative revision. We reviewed our experience with living related transplantation in pediatric recipients to examine the occurrence of biliary complications and the utility of percutaneous biliary procedures in their management. Over a 10-yr period, 48 living donor transplants were performed in 47 patients. Sixteen patients (33%) had biliary complications. Complications included 10 leaks (20%) and eight strictures (17%). Although leaks were treated predominantly with operation, other biliary complications were treated almost exclusively non-operatively. Self limited leaks that lead to biloma accumulation were most often treated via percutaneous catheter drainage and all strictures were treated using percutaneous transhepatic biliary cholangioplasty and stenting. Sixty-seven percent of biliary complications underwent non-operative biliary intervention. Most strictures were focal anastomotic strictures and were successfully treated with cholangioplasty although multiple interventions were necessary and patients required stenting for an average of 13 months. Three of eight strictures were diffuse in nature and these included the only patient who required retransplantation. Graft survival with respect to biliary complications was 94%; 1 yr, 5 yr and overall patient survival for those with biliary complications was 88, 88 and 81%, and for the entire living related group was 84, 81 and 77%, respectively. Although biliary complications are frequent in pediatric living related transplantation, they are not associated with decreased patient survival. Excepting significant bile leaks, the majority can be treated non-operatively via biliary cholangioplasty and stenting. Strictures are especially amenable to this technique which, in our experience, has been successful at decreasing or postponing the need for retransplantation.


Sanji Peiyuan decoction combined with dydrogesterone in the treatment of massive subchorionic hematoma: A case report.
Subchorionic Hematoma, often referred to as Bruce hematoma, can lead to serious obstetric complications such as intrauterine growth restriction and fetal death, as well as early and late pregnancy miscarriage, placental abruption, and premature rupture of membranes, posing great harm to both mother and fetus. At present, Western medical treatments have not shown satisfactory results, necessitating the discovery of more effective clinical treatment methods. Threatened miscarriage, Subchorionic hematoma, Iron deficiency anemia (mild). Sanji Peiyuan decoction combined with dydrogesterone. Following 17 days of treatment with Sanji Peiyuan decoction and oral dydrogesterone tablets, the hematoma was no longer detectable by ultrasound. The patient experienced no symptoms such as abdominal pain, bloating, or vaginal bleeding. She successfully gave birth around her due date, with both the mother and child in good health. The combination of Sanji Peiyuan decoction and oral dydrogesterone tablets shows promising clinical efficacy in treating Massive Subchorionic Hematomas. This method merits further clinical research.


How virulent are emerging maize-infecting mastreviruses?
Maize streak disease (MSD) is one of the most significant biotic constraints on the production of Africa's most important cereal crop. Until recently, the only virus known to cause severe MSD was the A-strain of maize streak virus (MSV/A), a member of the genus Mastrevirus, family Geminiviridae. However, over the past decade, two other mastreviruses, MSV/C and maize streak Réunion virus (MSRV), have been repeatedly found in the absence of MSV/A in maize plants displaying severe MSD symptoms. Here, we report on infectious clones of MSV/C and MSRV and test their ability to cause severe MSD symptoms. Although cloned MSV/C and MSRV genomes could cause systemic symptomatic infections in MSD-sensitive maize genotypes, these infections yielded substantially milder symptoms than those observed in the field. The MSV/C and MSRV isolates that we have examined are therefore unlikely to cause severe MSD on their own. Furthermore, mixed infections of MSRV and MSV/C with other mild MSV strains also consistently yielded mild MSD symptoms. It is noteworthy that MSRV produces distinctive striate symptoms in maize that are similar in pattern, albeit not in severity, to those seen in the field, showing that this virus may contribute to the severe MSD symptoms seen in the field. Therefore, despite not fulfilling Koch's postulates for MSV/C and MSRV as causal agents of severe MSD, we cannot exclude the possibility that these viruses could be contributing to currently emerging maize diseases.


Defining response in systemic lupus erythematosus: a study by the Systemic Lupus International Collaborating Clinics group.
In a preliminary attempt to develop a drug responder index for patients with systemic lupus erythematosus (SLE), 2 validated disease activity instruments were studied for their responsiveness and compared to a physician visual analog scale (VAS) assessment of disease activity. We attempted to determine whether these validated instruments were useful components in characterizing response in the setting of a clinical trial. Eighty paper patients were assessed using the British Isles Lupus Assessment Group (BILAG) and Systemic Lupus Disease Activity Index (SLEDAI) and by physician's assessment of global activity. The cases were arranged in random order and divided into groups of 20 patients and each group was assessed by 20 lupus experts; change in disease activity was recorded at 3 and 6 months compared to baseline using a physician VAS. Four different lupus experts assessed disease activity in all 80 patients at baseline and 3 and 6 months after initiation of therapy using the BILAG and SLEDAI instruments. BILAG and SLEDAI scores correlated well over time; however, in a regression analysis where average physician VAS were chosen as the outcome variable, a significant amount of variation in the average physician VAS not related to the SLEDAI and BILAG scores was noted. The physician VAS may be too blunt to assess response in SLE, because even among experienced lupus assessors, there were considerable differences in what influenced scoring decisions.


Palliative care consultations in patients with cancer: a mayo clinic 5-year review.
We sought to characterize the aggregate features and survival of patients who receive inpatient palliative care consultation, particularly focusing on patients with cancer, to identify opportunities to improve clinical outcomes. We reviewed prospectively collected data on patients seen by the Palliative Care Inpatient Consult Service at Mayo Clinic (Rochester, MN) from January 2003 to September 2008. Demographics, consultation characteristics, and survival were analyzed using Kaplan-Meier survival curves and Cox survival models. Cancer was the most common primary diagnosis (47%) in the 1,794 patients seen over the 5-year period. A significant growth in the annual number of palliative care consultations has been observed (113 in 2003 v 414 in 2007), despite stable total hospital admissions. Frequently encountered reasons for consultation included clarification of care goals (29%), assistance with dismissal planning (19%), and pain control (17%). Although patients with cancer had the highest median survival after consultation in this cohort versus patients with other diagnoses, we observed a 5-year trend of decreasing survival from admission to death and from consultation to death. Median time from admission to death for patients with cancer was 36 days in 2003 and only 19 days in 2008 (P < .01). Median time from consultation to death decreased from 33 days in 2003 to only 11.5 days in 2008 (P < .01). Patients with cancer often have complex needs that must be met within a short window for intervention. We highlight opportunities for improved multidisciplinary care for patients with advanced cancer and their families, including opportunity for earlier palliative care involvement, even in the outpatient setting.


Detecting the temporal structure of intermittent phase locking.
This study explores a method to characterize the temporal structure of intermittent phase locking in oscillatory systems. When an oscillatory system is in a weakly synchronized regime away from a synchronization threshold, it spends most of the time in parts of its phase space away from the synchronization state. Therefore characteristics of dynamics near this state (such as its stability properties and Lyapunov exponents or distributions of the durations of synchronized episodes) do not describe the system's dynamics for most of the time. We consider an approach to characterize the system dynamics in this case by exploring the relationship between the phases on each cycle of oscillations. If some overall level of phase locking is present, one can quantify when and for how long phase locking is lost, and how the system returns back to the phase-locked state. We consider several examples to illustrate this approach: coupled skewed tent maps, the stability of which can be evaluated analytically; coupled Rössler and Lorenz oscillators, undergoing through different intermittency types on the way to phase synchronization; and a more complex example of coupled neurons. We show that the obtained measures can describe the differences in the dynamics and temporal structure of synchronization and desynchronization events for the systems with a similar overall level of phase locking and similar stability of the synchronized state.


[Neurotransmitter stage in the development of peripheral part of autonomic nervous system].
The dynamics of neurotransmitter stage in the development of autonomic nervous system (ANS) was studied in rats starting from the moment of initial appearance of neurotransmitters acetylcholine and adrenalin in ANS peripheral part main nervous plexuses. Cryostat sections of embryos on developmental days 13.5, 16.5 and 18.5 and of neonatal rats aged 0.5 and 1.5 days were treated with glyoxilic acid, using Karnovsky-Roots method and impregnated with silver nitrate according to Bielschowsky-Gross method. Acetylcholinesterase in vagus and in spinal ganglia was found on day 13.5, while catecholamine in sympathetic trunc was detected in animals immediately after the birth. Periods of the beginning of neurotransmitter stage in ANS development in rat and man were compared and it was established that it was delayed in rat as in immature-born animal.


Re-stenosis of a sirolimus-coated stent in a heart transplant recipient with allograft vasculopathy.
We report the first case of in-stent re-stenosis in a sirolimus-coated stent in a heart transplant recipient with allograft vasculopathy. A 54-year-old male transplant recipient with new lesions in the left anterior descending artery detected during a routine surveillance catheterization was treated with two sirolimus-coated stents. Three months later he presented with dyspnea on exertion, and a repeat left heart catheterization demonstrated a focal area of in-stent stenosis in the proximal stent, which was treated with a cutting balloon.


The effect of different stretching techniques on ankle joint range of motion and dynamic postural stability after landing.
Stretching improves the flexibility of skeletal muscles, increases the range of motion (ROM) of joints. Stretching is important in prevention of sport-related injuries and influences muscle strength and performance. The effects of Static Stretching (SS) and Cyclic Stretching (CS) have been assessed by examining ROM, muscle power, vertical jump performance. However, the effects on postural control after CS compared with SS does not provide evidence for postural control after landing. The aim of this study was to examine the effects of stretching on the range of motion (ROM) of the ankle joint and dynamic postural stability upon landing on one leg. Twenty healthy subjects participated in this study. Participants were randomly assigned to SS, CS, and control conditions. The ankle was stretched in each condition for two minutes in a standing position. In the assessment of dynamic postural stability, the participant jumped and landed on one leg onto a force platform and the Dynamic Postural Stability Index (DPSI) was measured. Stability indices including those for medial-lateral, anterior-posterior, and vertical stability were calculated. The data were compared among 3 conditions with repeated measures ANOVA and the correlation between ankle range of motion (ROM), DPSI and the maximum vertical ground reaction force (vGRF max) were calculated. The results indicated that the ROM was significantly greater after SS and CS compared with the control condition. A significant improvement in DPSI was observed after CS. We showed that 2 minutes of CS had a positive influence on dynamic postural stability after landing on a single leg. Furthermore, CS may improve balance by increasing postural stability.


Single ascending oral dose pharmacokinetics and pharmacodynamics study of EV-077: the specific inhibitor of prostanoid- and isoprostane-induced cellular activation.
This study was performed to determine the oral pharmacokinetics (PK) of EV-077 and its effects on pharmacodynamic (PD) markers. EV-077 blocks prostanoid-induced and isoprostane-induced cellular activation, and is in development for the treatment of vascular inflammation and associated complications of type-2 diabetes.. This single-ascending-dose mono-centre study was randomised, placebo-controlled, and double-blinded within each dose group. Seven EV-077 doses were administered sequentially as an oral solution: 0.0125, 0.125, 0.375, 0.75, 1.25, 1.875 and 2.5 mg/kg body weight. PK, platelet aggregation, bleeding time and safety parameters were measured. Seven to eight healthy male subjects were dosed per group: five to six subjects received EV-077 and two subjects received placebo. Tmax was reached rapidly between 0.5 h and 1.0 h. Both Cmax and AUC increased linearly with the dose. The apparent terminal half-life (t½z) increased with the dose, most likely reflecting the increasing last quantifiable concentration with increasing dose; at 2.5 mg/kg, it was 2.7-6.9 h. Measurement of platelet aggregation showed no effect at 0.0125 mg/kg, and a full and reversible inhibition at doses of 0.125-2.5 mg/kg. The average bleeding time was dose-dependently prolonged, but was always below 9 min. The PK/PD profile showed that at plasma concentrations above 20 ng/ml, EV-077 platelet aggregation was completely inhibited (>90 %). All tested doses were well tolerated. Orally administered EV-077 was well tolerated, readily absorbed, reached Cmax within 1 h, with a linear PK based on Cmax and AUC. The inhibition of platelet aggregation was complete and reversible at doses of 0.125 mg/kg and higher, and average bleeding time was below 9 min.


Toxigenic Escherichia coli with high antibiotic resistance index recovered from sands of recreational beaches of Mumbai, India.
Mumbai, India's seven-island city, is known for its sandy beaches as a major tourist attraction, but urbanization and industrialization have weakened the environment. Unregulated sewage disposal and untreated effluents off the coast have made the beach environment vulnerable. Therefore, monitoring water and sand quality at beaches should be mandatory. This study was thus designed to determine the microbiological status of selected sandy beaches, viz. Versova, Juhu, and Girgaon. The study found fecal coliforms in the sand, with stx1 and stx2 genes specific for Shiga toxin-producing E. coli pathotypes in 5.5 % of isolates, whereas the presence of eaeA gene specific for enteropathogenic E. coli pathotype was detected in 12.2 % of isolates, and the presence of the LT and ST genes specific for enterotoxigenic E. coli pathotype was detected in 6.6 % of isolates. Multiple antibiotic-resistant indices indicated high-risk contamination sources. The study suggests routine monitoring of pollution levels at coastal cities' beaches.


Immunotherapy with commercial venoms is efficacious for anaphylactic reactions to Vespa orientalis stings.
Vespa orientalis (VO) stings occasionally induce anaphylaxis. In the absence of commercial VO venom, allergists use commercial venoms for immunotherapy, despite having no indication regarding efficacy. We attempted to examine the effectiveness of immunotherapy with commercial venoms in patients with VO allergy and to identify the venom accountable for this effect. Patients who unequivocally identified VO as the culprit insect were administered venom immunotherapy with the commercial venoms available in Israel to which they had positive skin tests. Patients were also skin tested with VO venom sac extracts and, after reaching the maintenance dose, were sting challenged by a live insect. The allergenic components in the venom were determined by immunoblotting. Twelve patients were recruited and, based on their skin test results, all were treated with yellow jacket (YJ) venom, either alone or combined with other venoms. All 8 patients who were sting challenged by VO demonstrated positive skin test responses to VO venom. Six of the stung patients tolerated the sting challenge uneventfully. Two patients developed minimal transient symptoms that resolved spontaneously. SDS-PAGE with patient sera suggested cross-reactivity between VO and YJ venoms at molecular weights of 39-42 kDa. Using phospholipases, antigen 5 and hyaluronidase derived from several Vespa, Dolichovespula and Vespula species, hyaluronidase is possibly accountable for inducing the allergic reaction. In the absence of commercial VO venom the practice of treating patients allergic to this insect with available commercial venoms seems to be efficacious and YJ venom is probably responsible for this effect.


Two new 14, 15-secopregnane-type steroidal glycosides from the roots of Cynanchum limprichtii.
Two new steroidal glycosides 1 and 2, along with three known ones (3-5), were isolated from the 95% ethanol extract of the roots of Cynanchum limprichtii Schltr. The structure of the new compounds was elucidated as 3-O-α-L-diginopyranosyl-(1→4)-β-D-digitoxopyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-thevetopyranosyl-14, 16:15, 20:18, 20-triepoxy-14, 15-secopregn-4, 6, 8 (14)-triene (1) and 3-O-α-L-cymaropyranosyl-(1→4)-β-D-digitoxopyranosyl- (1→4)-β-D-3-demethyl-2-deoxythevetopyranosyl-14, 16: 15, 20: 8, 20-triepoxy-14, 15-secopregn-5, 8 (14)-diene (2) on the basis of spectroscopic analysis together with acidic hydrolysis. All compounds showed cytotoxic activity against the human cancer cell line HL60, with IC50 values of 55.36, 65.41, 17.88, 17.68 and 33.5 μM, respectively. While, only compound 3 showed cytotoxicity against the Caco-2 cell line, with an IC50 value of 67.47 μM.


Cigarette smoking and the risk of cervical neoplasia.
The relationship between cigarette smoking and risk of cervical neoplasia was evaluated in a case-control study of 183 women with cervical intraepithelial neoplasia compared with 183 age-matched outpatient controls, and of 230 cases of invasive cervical cancer compared with 230 controls in hospital for acute conditions unrelated to any of the identified or suspected risk factors for cervical cancer. Current cigarette smoking was associated with an elevated risk of cervical intraepithelial neoplasia (relative risk = 1.76, 95 per cent confidence interval = 1.14-2.27) and of invasive cancer (relative risk = 1.69, 95 per cent confidence interval = 1.08-2.65). This association was only partially accounted for by a large number of identified potential confounding factors, including indicators of socioeconomic status and sexual habits. The risk increased with the number of cigarettes smoked and was apparently greater for women who started smoking at younger ages. The relative risk of intraepithelial neoplasia was elevated within 20 years after the start of smoking and showed little tendency to increase with increasing duration. On the other hand, the risk of invasive cervical cancer was apparently unaffected by smoking less than 20 years and increased steadily thereafter, reaching a point estimate of 3.63 after 40 years or more. If one assumes that intraepithelial neoplasia is an early stage of cervical cancer, this pattern of risk is consistent with the predictions from the multistage theory of carcinogenesis, if the effect of smoking is on one of the earlier stages. No obvious distorting factors, apart from the play of chance, is likely to produce such a risk pattern.


Therapeutic experience with fludrocortisone in diabetic postural hypotension.
Fourteen patients with diabetic postural hypotension were treated with fludrocortisone for a mean 12 months (range 6-30 months). The mean daily dose of fludrocortisone was 0.2 mg (range 0.1 mg-0.4 mg). Standing systolic and diastolic blood pressures increased significantly (P less than 0.001) after treatment with fludrocortisone and the postural hypotension decreased significantly (P less than 0.001). Thirteen patients noted considerable symptomatic improvement. Fludrocortisone should be used cautiously in patients with congestive cardiac failure or the nephrotic syndrome.


New Benzimidazole-1,2,4-Triazole Hybrid Compounds: Synthesis, Anticandidal Activity and Cytotoxicity Evaluation.
Owing to the growing need for antifungal agents, we synthesized a new series 2-((5-(4-(5-substituted-1H-benzimidazol-2-yl)phenyl)-4-substituted-4H-1,2,4-triazol-3-yl)thio)-1-(substitutedphenyl)ethan-1-one derivatives, which were tested against Candida species. The synthesized compounds were characterized and elucidated by FT-IR, ¹H-NMR, 13C-NMR and HR-MS spectroscopies. The synthesized compounds were screened in vitro anticandidal activity against Candida species by broth microdiluation methods. In vitro cytotoxic effects of the final compounds were determined by MTT assay. Microbiological studies revealed that compounds 5m, 5o, 5r, 5t, 5y, 5ab, and 5ad possess a good antifungal profile. Compounds 5w was the most active derivative and showed comparable antifungal activity to those of reference drugs ketoconazole and fluconazole. Cytotoxicity evaluation of compounds 5m, 5o, 5r, 5w, 5y, 5ab and 5ad showed that compounds 5w and 5ad were the least cytotoxic agents. Effects of these two compounds against ergosterol biosynthesis were observed by LC-MS-MS method, which is based on quantification of ergosterol level in C. albicans. Compounds 5w and 5d inhibited ergosterol biosynthesis concentration dependently. A fluorescence microscopy study was performed to visualize effect of compound 5w against C. albicans at cellular level. It was determined that compound 5w has a membrane damaging effect, which may be related with inhibition of biosynthesis of ergosterol.


Remnants of an ancient pathway to L-phenylalanine and L-tyrosine in enteric bacteria: evolutionary implications and biotechnological impact.
The pathway construction for biosynthesis of aromatic amino acids in Escherichia coli is atypical of the phylogenetic subdivision of gram-negative bacteria to which it belongs (R. A. Jensen, Mol. Biol. Evol. 2:92-108, 1985). Related organisms possess second pathways to phenylalanine and tyrosine which depend upon the expression of a monofunctional chorismate mutase (CM-F) and cyclohexadienyl dehydratase (CDT). Some enteric bacteria, unlike E. coli, possess either CM-F or CDT. These essentially cryptic remnants of an ancestral pathway can be a latent source of biochemical potential under certain conditions. As one example of advantageous biochemical potential, the presence of CM-F in Salmonella typhimurium increases the capacity for prephenate accumulation in a tyrA auxotroph. We report the finding that a significant fraction of the latter prephenate is transaminated to L-arogenate. The tyrA19 mutant is now the organism of choice for isolation of L-arogenate, uncomplicated by the presence of other cyclohexadienyl products coaccumulated by a Neurospora crassa mutant that had previously served as the prime biological source of L-arogenate. Prephenate aminotransferase activity was not conferred by a discrete enzyme, but rather was found to be synonymous with the combined activities of aspartate aminotransferase (aspC), aromatic aminotransferase (tyrB), and branched-chain aminotransferase (ilvE). This conclusion was confirmed by results obtained with combinations of aspC-, tyrB-, and ilvE-deficient mutations in E. coli. An example of disadvantageous biochemical potential is the presence of a cryptic CDT in Klebsiella pneumoniae, where a mutant carrying multiple enzyme blocks is the standard organism used for accumulation and isolation of chorismate.(ABSTRACT TRUNCATED AT 250 WORDS)


[Interactive determination of the parameters of mathematical models in planning radiotherapy of malignant tumors. 3. Method of local adjustment of the parameters of mathematical models (examples of application)].
To increase the accuracy of calculation of tolerance doses of the likelihood of radiation-induced complications in normal organs and tissues by using mathematical models, the author has developed a method for local mathematical model parameter adjustment (LMMPA) which included analysis of the structure of a mathematical model, systematization of clinical information and its goal-oriented use to determine the parameters of a model. The necessity of developing the LMMPA method stemmed from the complexity of the body exposed to radiation a system and from the quest for taking into account the impact of the system on the values of mathematical models. LMMPA may be regarded as the extension of determination of the parameters of mathematical models or as the interactive determination of their parameters that describing radiation exposures of complex biological systems. Different aspects of using of LMMPA are indicated how to apply it to the determination of tolerance doses for connective tissue, lung tissue, and the brain by using the Ellis and LQ models. The extended LMMPA is shown how to determine the parameters of a mathematical models for calculation of the likelihood of radiation-induced complications in the lung tissue.


Evidence for broad versus segregated projections from cholinergic and noradrenergic nuclei to functionally and anatomically discrete subregions of prefrontal cortex.
The prefrontal cortex (PFC) is implicated in a variety of cognitive and executive operations. However, this region is not a single functional unit; rather, it is composed of several functionally and anatomically distinct networks, including anterior cingulate cortex (ACC), medial prefrontal cortex (mPFC), and orbitofrontal cortex (OFC). These prefrontal subregions serve dissociable behavioral functions, and are unique in their afferent and efferent connections. Each of these subregions is innervated by ascending cholinergic and noradrenergic systems, each of which likewise has a distinct role in cognitive function; yet the distribution and projection patterns of cells in the source nuclei for these pathways have not been examined in great detail. In this study, fluorescent retrograde tracers were injected into ACC, mPFC, and OFC, and labeled cells were identified in the cholinergic nucleus basalis of Meynert (NBM) and noradrenergic nucleus locus coeruleus (LC). Injections into all three cortical regions consistently labeled cells primarily ipsilateral to the injection site with a minimal contralateral component. In NBM, retrogradely labeled neurons were scattered throughout the rostral half of the nucleus, whereas those in LC tended to cluster in the core of the nucleus, and were rarely localized within the rostral or caudal poles. In NBM, more than half of all retrogradely labeled cells possessed axon collaterals projecting two or more PFC subregions. In LC, however, only 4.3% of retrogradely labeled neurons possessed collaterals targeting any two prefrontal subregions simultaneously, and no cells were identified that projected to all three regions. Of all labeled LC neurons, 49.3% projected only to mPFC, 28.5% projected only to OFC, and 18.0% projected only to ACC. These findings suggest that subsets of LC neurons may be capable of modulating neuronal activity in individual prefrontal subregions independently, whereas assemblies of NBM cells may exert a more unified influence on the three areas, simultaneously. This work emphasizes unique aspects of the cholinergic and noradrenergic projections to functionally and anatomically distinct subregions of PFC and provides insights regarding global versus segregated regulation of prefrontal operations by these neuromodulatory pathways.


Prevalence of microorganisms in dead mink kits from Aleutian-disease-infected and non-infected farms.
Bacterial and fungi were isolated from different tissues (brain, liver, heart) taken from 81 dead newborn mink originating from Aleutian disease (AD) infected and AD-non-infected farms. Of the 123 isolates obtained, 96% were bacterial isolates (predominantly Gram-negative) and 4% were fungi. The prevalence of microorganisms appeared less common in kits from AD-non-infected farms (55%) than from AD-infected farms (73%), although the difference was not significant. The liver was the most highly infected site in both groups and generally was only infected by one microorganism species. Proteus spp (23%), Escherichia coli (16%), Staphylococcus aureus (11%) and Enterobacter cloacae (9%) were the most frequently isolated germs. These findings are similar to those of other studies but the role of these microorganisms as specific pathogens or secondary invaders remains controversial.


A plant caspase-like protease activated during the hypersensitive response.
To test the hypothesis that caspase-like proteases exist and are critically involved in the implementation of programmed cell death (PCD) in plants, a search was undertaken for plant caspases activated during the N gene-mediated hypersensitive response (HR; a form of pathogen-induced PCD in plants) in tobacco plants infected with Tobacco mosaic virus (TMV). For detection, characterization, and partial purification of a tobacco caspase, the Agrobacterium tumefaciens VirD2 protein, shown here to be cleaved specifically at two sites (TATD and GEQD) by human caspase-3, was used as a target. In tobacco leaves, specific proteolytic processing of the ectopically produced VirD2 derivatives at these sites was found to occur early in the course of the HR triggered by TMV. A proteolytic activity capable of specifically cleaving the model substrate at TATD was partially purified from these leaves. A tetrapeptide aldehyde designed and synthesized on the basis of the elucidated plant caspase cleavage site prevented fragmentation of the substrate protein by plant and human caspases in vitro and counteracted TMV-triggered HR in vivo. Therefore, our data provide a characterization of caspase-specific protein fragmentation in apoptotic plant cells, with implications for the importance of such activity in the implementation of plant PCD.


[Induced radioactivities in treatment head of 10 MV X-ray linear Accelerator (author's transl)].
The concentration of 60Co and stable cobalt in the shellfishes and seaweeds which were collected from the coast around Japan were obtained by low background beta counting for 60Co and spectrometric determination of stable cobalt. The results obtained were as follows: (1) The 60Co was found in the shellfishes which were enriched in the stable cobalt. (2) The correlation between 60Co and stable cobalt was rather good in the shellfishes. (3) The activities of 60Co was found in the seaweeds taken from seashore near the nuclear power plant. (4) There was great differences in the specific activities between samples collected from contaminated and not contaminated area.


High expression of cyclin B1 predicts a favorable outcome in patients with follicular lymphoma.
Substantial research has been dedicated to the study of the relationship between genetic mechanisms regulating cell functions in tumors and how those tumors respond to various treatment regimens. Because these mechanisms are still not well understood, we have chosen to study the genetic makeup of 57 tumor samples from patients with follicular lymphoma (FL). Our goal was to develop a prognostic tool, which can be used as an aid in determining FL patients with tumors genetically predisposed to a successful treatment with the CHOP (cyclophosphamide, vincristine, doxorubicin, prednisone) regimen. To select relevant genes, high-density oligonucleotide arrays were used. There were 14 genes highly expressed in FL patients that responded well to CHOP chemotherapy, and 11 of these were involved in G2/M transition of the cell cycle, in mitosis, or in DNA modulation. A high expression of CCNB1 (cyclin B1), CDC2, CDKN3A, CKS1B, ANP32E, and KIAA0101, but not of the proliferation-related antigen Ki-67, was associated with better survival rate in a univariate analysis. CCNB1 expression had an independent prognostic value when included in a multivariate analysis together with the 5 parameters of the follicular lymphoma international prognostic index.


Local persistence of Mann's soft-haired mouse Abrothrix manni (Rodentia, Sigmodontinae) during Quaternary glaciations in southern Chile.
Quaternary climatic oscillations have impacted Patagonian sigmodontine fauna, leaving traceable genetic footprints. In southern Chile, changes in the landscape included transitions to different vegetation formations as well as the extension of ice sheets. In this study, we focus on the Valdivian forest endemic and recently described sigmodontine species Abrothrix manni. We aim to assess the genetic structure of this species, testing for the existence of intraspecific lineages, and inferring the recent demographic history of the species. Analyses were based on the first 801 bp of the mitochondrial gene Cytocrhome-b from 49 individuals of A. manni collected at 10 localities that covers most part of its geographic distribution. Genealogical analyses recovered two main intraspecific lineages that are geographically segregated and present an intermediate site of secondary contact. Historical demography shows signal of recent population decrease. Based on these results, we proposed that current genetic diversity of A. manni differentiated in at least two distinct refugial areas in southern Chile. This scenario, in addition to be unique among those uncovered for the so far studied Valdivian forest rodents, is noteworthy because of the reduced geographic scale inhabited by the species.


[Liver damage associated with the combination drug amoxicillin-clavulanic acid (Augmentin)].
To describe the case histories of patients with liver damage associated with the antibacterial combination amoxicillin-clavulanic acid. Descriptive. University Institute for General Practice, Rotterdam, the Netherlands. All reports (n = 40) of hepatic injury, attributed to amoxicillin-clavulanic acid (causal relationship 'possible', 'probable' or 'certain') and received by the Drug Safety Unit of the Inspectorate for Health Care since the registration of the combination (1982) until August 1996 were described and evaluated. The reports concerned 28 men and 12 women with an average age of 61 years who had used the drug mostly for a respiratory tract infection. The latency period between first in-take and onset of the reaction was 3 weeks on average. The mean duration of hepatic injury was approximately 6 weeks. The liver injury resolved fully. Five patients had had the reaction on more than one occasion. The main symptoms were jaundice, nausea, pruritus and abdominal pain. The pattern of hepatic injury was mostly cholestatic or mixed hepatocellular-cholestatic. Since liver damage to amoxicillin-clavulanic acid appears to be infrequent, this adverse effect is probably not caused by amoxicillin alone. The risk of liver damage to amoxicillin-clavulanic acid is highest in elderly patients treated with the combination on several occasions. Doctors should restrict the use of this combination to the treatment of infections with amoxicillin-resistant bacteria.


[The assessment of mechanical heart valves stenosis in adults after aortic valve replacement: the advantage of full-flow design of mechanical valve].
The analysis of transprosthetic hemodynamics in adults after aortic valve replacement in the Bakoulev Center for Cardiovascular Surgery in 2007-2010 demonstrated the hemodynamic advantage of the concept of new full-flow mechanical aortic valve prosthesis "CorBeat". Having the same size of internal orifice and tissue annulus diameters, the values of transprosthetic parameters (peak and mean gradients, blood flow velocities) through "CorBeat" were close to physiological values of transvalvular native aortic parameters and had a tendency to be not dependent on the size of prosthesis (p = 0.63). In the article for the first time a morphometric database of geometric values of internal orifice area of normal native aortic valves in adults was used taking into account both the gender and the body surface area's of a patient. There was also used the standardized prosthesis size Z-score which represents the number of SDs by which the internal prosthesis area differs from the mean normal native aortic valve area for the patient's body surface area. The article emphasizes the need of the personal selection of the size and the type of prosthesis for any patient as well as the need for new design development of prosthetic heart valves.


Lethal sectoring and the delayed induction of aneuploidy in yeast.
Persistent lethal sectoring in a homothallic strain of yeast has been ascribed to tetrasomy for chromosome I. Such aneuploids can appear many generations after irradiation. The data thus indicate that an induced predisposition towards aneuploidy can be prolonged through successive post-irradiation cell divisions. Sporadic cell death in tetrasomics for chromosome I was found to result from a metabolic imbalance and not from a genetic instability conseqent to aneuploidy. This imbalance may be due to a dosage effect involving cistrons for ribosomal RNA since many of these are known to be located on chromosome I. Tetrasomy is not the only cause of persistent lethal sectoring; the phenomenon has been initiated through genetic recombination involving normal diploids. It has also been concluded that, in trisomics, equational division of the supernumerary chromosome sometimes occurs at the first meiotic division.


Atlanto-axial rotatory fixation: a serious potential complication of paediatric ENT surgery that requires prompt diagnosis and treatment.
Atlanto-axial rotatory fixation is a persistent deformity of the C1-2 vertebral relationship caused by subluxation of the articular surfaces, and can occur after positioning for ENT procedures where the head is rotated - for example to access the ear or posterior triangle of the neck. If promptly recognised, it can usually be managed successfully with conservative methods, without long-lasting sequelae, but delayed or inappropriate management may lead to permanent neck deformity, neurological problems and pain. Case review. Two children with atlanto-axial rotatory fixation following ENT surgery; one child was referred early and managed successfully, and one had delayed referral resulting in permanent severe positional deformity. Atlanto-axial rotatory fixation is easily missed; there are significant clinical and medicolegal implications if it is not promptly recognised. A suggested management algorithm is presented.


Pathogenesis of acute avian malaria. II. Anemia mediated by a cold-active autohemagglutinin from the blood of chickens with acute Plasmodium gallinaceum infection.
A cold-active hemagglutinin for trypsinized human type "O" erythrocytes (CAH) from blood of chickens with acute Plasmodium gallinaceum malaria was found to be associated with 19 S and 7 S globulin fractions of malarious chicken blood, but cleavage with 2-mercaptoethanol indicated that it was primarily of the IgM class of antibody. In serologic tests CAH reacted with trypsinized erythrocytes, and anti-chicken globulin. It did not react with other of the antigens or antibodies detected in the blood of malarious chickens. When the absorbed and eluted CAH was injected into normal chickens it produced an anaphylactic-like shock and caused a 25% reduction in red blood cell counts within 48 hours. Plasma samples collected during this interval showed signs of hemolysis. Reactions of blood cells from the recipient birds with fluorescein conjugated anti-chicken globulin indicated that CAH reacted with erythrocytes. The absence of fluorescent activity 3 days after injection suggested that these erythrocytes had been removed from the circulation. When normal chickens were injected with trypsinized autologous blood cells, CAH was detected within 3 days. The agglutination test again was active at temperatures below 22 degrees C and was negative when tested at 37 degrees C. In these birds the appearance of CAH was accompanied by reductions in red blood cell counts and by hemolysis. The results of these experiments suggest that CAH was not stimulated by plasmodial parasite antigen, but rather by autoantigens, which appear to be common to heterologous animal species, and which were in some manner expressed by the presence of the intracellular parasites, or by trypsin treatment. The experiments further suggest that this autohemagglutinin was partially causal of malarial anemia. The presence of other anemia factor(s) was indicated by anemia following injection of plasma that had been absorbed free of CAH.


Direct modulation of microtubule stability contributes to anthracene general anesthesia.
Recently, we identified 1-aminoanthracene as a fluorescent general anesthetic. To investigate the mechanism of action, a photoactive analogue, 1-azidoanthracene, was synthesized. Administration of 1-azidoanthracene to albino stage 40-47 tadpoles was found to immobilize animals upon near-UV irradiation of the forebrain region. The immobilization was often reversible, but it was characterized by a longer duration consistent with covalent attachment of the ligand to functionally important targets. IEF/SDS-PAGE examination of irradiated tadpole brain homogenate revealed labeled protein, identified by mass spectrometry as β-tubulin. In vitro assays with aminoanthracene-cross-linked tubulin indicated inhibition of microtubule polymerization, similar to colchicine. Tandem mass spectrometry confirmed anthracene binding near the colchicine site. Stage 40-47 tadpoles were also incubated 1 h with microtubule stabilizing agents, epothilone D or discodermolide, followed by dosing with 1-aminoanthracene. The effective concentration of 1-aminoanthracene required to immobilize the tadpoles was significantly increased in the presence of either microtubule stabilizing agent. Epothilone D similarly mitigated the effects of a clinical neurosteroid general anesthetic, allopregnanolone, believed to occupy the colchicine site in tubulin. We conclude that neuronal microtubules are "on-pathway" targets for anthracene general anesthetics and may also represent functional targets for some neurosteroid general anesthetics.


Profiling peripheral nerve macrophages reveals two macrophage subsets with distinct localization, transcriptome and response to injury.
While CNS microglia have been extensively studied, relatively little is known about macrophages populating the peripheral nervous system. Here we performed ontogenic, transcriptomic and spatial characterization of sciatic nerve macrophages (snMacs). Using multiple fate-mapping systems, we show that snMacs do not derive from the early embryonic precursors colonizing the CNS, but originate primarily from late embryonic precursors and become replaced by bone-marrow-derived macrophages over time. Using single-cell transcriptomics, we identified a tissue-specific core signature of snMacs and two spatially separated snMacs: Relmα+Mgl1+ snMacs in the epineurium and Relmα-Mgl1- snMacs in the endoneurium. Globally, snMacs lack most of the core signature genes of microglia, with only the endoneurial subset expressing a restricted number of these genes. In response to nerve injury, the two resident snMac populations respond differently. Moreover, and unlike in the CNS, monocyte-derived macrophages that develop during injury can engraft efficiently in the pool of resident peripheral nervous system macrophages.


Personalized next-best action recommendation with multi-party interaction learning for automated decision-making.
Automated next-best action recommendation for each customer in a sequential, dynamic and interactive context has been widely needed in natural, social and business decision-making. Personalized next-best action recommendation must involve past, current and future customer demographics and circumstances (states) and behaviors, long-range sequential interactions between customers and decision-makers, multi-sequence interactions between states, behaviors and actions, and their reactions to their counterpart's actions. No existing modeling theories and tools, including Markovian decision processes, user and behavior modeling, deep sequential modeling, and personalized sequential recommendation, can quantify such complex decision-making on a personal level. We take a data-driven approach to learn the next-best actions for personalized decision-making by a reinforced coupled recurrent neural network (CRN). CRN represents multiple coupled dynamic sequences of a customer's historical and current states, responses to decision-makers' actions, decision rewards to actions, and learns long-term multi-sequence interactions between parties (customer and decision-maker). Next-best actions are then recommended on each customer at a time point to change their state for an optimal decision-making objective. Our study demonstrates the potential of personalized deep learning of multi-sequence interactions and automated dynamic intervention for personalized decision-making in complex systems.


Redox-regulated signaling by lactosylceramide in the proliferation of human aortic smooth muscle cells.
Previously, our laboratory reported that lactosylceramide (LacCer) stimulated human aortic smooth muscle cell proliferation via specific activation of p44 mitogen-activated protein kinase (MAPK) in the p21(ras)/Raf-1/MEK2 pathway and induced expression of the transcription factor c-fos downstream to the p44 MAPK signaling cascade (Bhunia A. K., Han, H., Snowden, A., and Chatterjee S. (1996) J. Biol. Chem. 271, 10660-10666). In the present study, we explored the role of free oxygen radicals in LacCer-mediated induction of cell proliferation. Superoxide levels were measured by the lucigenin chemiluminescence method, MAPK activity was measured by immunocomplex kinase assays, and Western blot analysis and c-fos expression were measured by Northern blot assay. We found that LacCer (10 microM) stimulates endogenous superoxide production (7-fold compared with control) in human aortic smooth muscle cells specifically by activating membrane-associated NADPH oxidase, but not NADH or xanthine oxidase. This process was inhibited by an inhibitor of NADPH oxidase, diphenylene iodonium (DPI), and by antioxidants, N-acetyl-L-cysteine (NAC) or pyrrolidine dithiocarbamate. NAC and DPI both abrogated individual steps in the signaling pathway leading to cell proliferation. For example, the p21(ras).GTP loading, p44 MAPK activity, and induction of transcription factor c-fos all were inhibited by NAC and DPI as well as an antioxidant pyrrolidine dithiocarbamate or reduced glutathione (GSH). In contrast, depletion of GSH by L-buthionine (S, R)-sulfoximine up-regulated the above described signaling cascade. In sum, LacCer, by virtue of activating NADPH oxidase, produces superoxide (a redox stress signaling molecule), which mediates cell proliferation via activation of the kinase cascade. Our findings may explain the potential role of LacCer in the pathogenesis of atherosclerosis involving the proliferation of aortic smooth muscle cells.


The extraneural and intraneural arterial anatomy of the ulnar nerve at the elbow.
The purpose of this report was to investigate the vascular anatomy of the ulnar nerve at the elbow with a technique of combined India ink and latex injection followed by nondissection chemical debridement. Twenty-two fresh human cadaveric arms were injected with India ink to stain the intraneural microcirculation followed immediately by latex injection and chemical debridement for study of the extraneural vascularization. After clearing with a modified Spalteholtz technique, the intraneural blood supply was studied in 10 of the specimens. The findings demonstrated a consistent but segmental extraneural and intraneural vascular supply from the superior ulnar collateral, inferior ulnar collateral, and posterior ulnar recurrent arteries. No identifiable direct anastomosis was seen between the superior ulnar collateral and posterior ulnar recurrent arteries in 20 of 22 arms. The inferior ulnar collateral artery provided the only direct vascularization to the nerve in the region just proximal to the cubital tunnel. Although the clinical importance of maintaining specific arterial sources to the ulnar nerve has not been determined, these anatomic findings indicate that the arterial contribution from the inferior ulnar collateral artery may be more important than appreciated previously.


Recruiting Nurse Participants in Ambulatory Care Nursing Research.
Patient care needs in ambulatory care (AC) settings continue to grow and evolve in the United States, with commensurate growth of nursing responsibilities in AC. Conducting research on the nursing workforce and nursing practice is essential to understanding and meeting the needs of nurses and patients in this setting. However, the structures and characteristics of AC settings pose challenges for conducting research on AC nursing practice. This article explains unique barriers to participation in research for nurses in AC, describes recruitment challenges for nurse researchers in AC, and provides strategies to increase recruitment of nurses for AC research. Researchers in AC must find ways to recruit representative participant samples, be clear and precise in defining terms, and report robust demographic information about participants and their practice settings.