Evaluation of hexadecyloxypropyl-9-R-[2-(Phosphonomethoxy)propyl]- adenine, CMX157, as a potential treatment for human immunodeficiency virus type 1 and hepatitis B virus infections.
9-R-[2-(Phosphonomethoxy)propyl]-adenine (tenofovir) is an acyclic nucleoside phosphonate with antiviral activity against human immunodeficiency virus type 1 (HIV-1) and hepatitis B virus (HBV). Tenofovir is not orally bioavailable but becomes orally active against HIV-1 infection as the disoproxil ester (tenofovir disoproxil fumarate [Viread]). We have developed an alternative strategy for promoting the oral availability of nucleoside phosphonate analogs which involves esterification with a lipid to form a lysolecithin mimic. This mimic can utilize natural lysolecithin uptake pathways in the gut, resulting in high oral availability. Since the mimic is not subject to cleavage in the plasma by nonspecific esterases, it remains intact in the circulation and facilitates uptake by target cells. Significant drops in apparent antiviral 50% effective concentrations (EC(50)s) of up to 3 logs have been observed in comparison with non-lipid-conjugated parent compounds in target cells. We have applied this technology to tenofovir with the goal of increasing oral availability, decreasing the apparent EC(50), and decreasing the potential for nephrotoxicity by reducing the exposure of the kidney to the free dianionic tenofovir. Here we report that, in vitro, the hexadecyloxypropyl ester of tenofovir, CMX157, is 267-fold more active than tenofovir against HIV-1 and 4.5-fold more active against HBV. CMX157 is orally available and has no apparent toxicity when given orally to rats for 7 days at doses of 10, 30, or 100 mg/kg/day. Consequently, CMX157 represents a second-generation tenofovir analog which may have an improved clinical profile.


Exploiting quorum sensing to confuse bacterial pathogens.
Cell-cell communication, or quorum sensing, is a widespread phenomenon in bacteria that is used to coordinate gene expression among local populations. Its use by bacterial pathogens to regulate genes that promote invasion, defense, and spread has been particularly well documented. With the ongoing emergence of antibiotic-resistant pathogens, there is a current need for development of alternative therapeutic strategies. An antivirulence approach by which quorum sensing is impeded has caught on as a viable means to manipulate bacterial processes, especially pathogenic traits that are harmful to human and animal health and agricultural productivity. The identification and development of chemical compounds and enzymes that facilitate quorum-sensing inhibition (QSI) by targeting signaling molecules, signal biogenesis, or signal detection are reviewed here. Overall, the evidence suggests that QSI therapy may be efficacious against some, but not necessarily all, bacterial pathogens, and several failures and ongoing concerns that may steer future studies in productive directions are discussed. Nevertheless, various QSI successes have rightfully perpetuated excitement surrounding new potential therapies, and this review highlights promising QSI leads in disrupting pathogenesis in both plants and animals.


Hepatoid adenocarcinoma of the peritoneal cavity: Prolonged survival after debulking surgery and 5-fluorouracil, leucovorin and oxaliplatin (FOLFOX) therapy.
Hepatoid adenocarcinoma is an alpha-fetoprotein producing adenocarcinoma arising in numerous extra-hepatic organs. The diagnosis may be difficult because of the varied presentation but immunohistochemistries help make the diagnosis. The prognosis is often poor but in this report, we present a young female with hepatoid carcinoma who is doing well more than three years after her diagnosis.


Evaluation of diagnostic tests using information theory for multi-class diagnostic problems and its application for the detection of occlusal caries lesions.
Several methods are available to evaluate the performance of the tests when the purpose of the diagnostic test is to discriminate between two possible disease states. However multi-class diagnostic problems frequently appear in many areas of medical science. Hence, there is a need for methods which will enable us to characterize the accuracy of diagnostic tests when there are more than two possible disease states. To show that two information theory measures, information content (IC) and proportional reduction in diagnostic uncertainty (PRDU), can be used for the evaluation of the performance of diagnostic tests for multi-class diagnostic problems that may appear in different areas of medical science. Diagnostic accuracy study. Sixty freshly extracted permanent human molar and pre-molar teeth suspected to have occlusal caries lesions were selected for the study and were assessed by two experienced examiners. Each examiner performed two evaluations. Histological examination was used as the gold standard. The scores of the histological examination were defined as sound (n=11), enamel caries (n=22) and dentin caries (n=27). Diagnostic performance of i) visual inspection, ii) radiography, iii) laser fluorescence (LF) and iv) micro-computed tomography (M-CT) caries detection methods was evaluated by calculating IC and PRDU. Micro-computed tomography examination was the best method among the diagnostic techniques for the diagnosis of occlusal caries in terms of both IC and PRDU. M-CT examination supplied the maximum diagnostic information about the diagnosis of occlusal caries in the first (IC: 1.056; p<0.05), (PRDU: 70.5%) and second evaluation (IC: 1.105; p<0.05), (PRDU: 73.8%) for the first examiner. M-CT examination was the best method among the diagnostic techniques for the second examiner in both the first (IC:1.105; p<0.05), (PRDU:73.8%) and second evaluation (IC:1.061; p<0.05), (PRDU:70.8%). IC and PRDU were lowest for visual inspection. The present study demonstrates that IC and PRDU can be used to evaluate diagnostic test performance when multiple disease states are being evaluated.


Quality of sleep in patients with Parkinson's disease.
Parkinson's disease (PD) is a progressive neurodegenerative disorder causing motor and non-motor symptoms. The latter are common and include autonomic dysfunction, cognitive impairment, and sleep difficulties. Many of the non-motor aspects of PD such as sleep disturbance are more common and significantly affect the day-to-day activities of patients and their quality of life. The most important aim of this study was to evaluate the sleep quality in patients with PD. This case-control study was performed on patients with PD referred to the Neurology Clinic of our teaching hospital in 2011. Thirty-four patients with PD and 34 healthy people as control group were enrolled in this study. Sleep quality of patients and control was evaluated by Parkinson's disease sleep scale (PDSS) questionnaire. PDSS is a reliable and valid tool to measure sleep disorders in PD. The mean total PDSS score in patient group was 55.29 (SD = 26.92) indicating moderate to severe sleep disturbances whereas, the mean total score in control group was 20.34 (SD = 10.65). Difference between the two groups' mean scores was significant (P < 0.05). Our study demonstrated that patients with PD experienced poorer nocturnal sleep quality than the control group.


Functional halt positions of rotary FOF1-ATPase correlated with crystal structures.
The F(O)F(1)-ATPase is a rotary molecular motor. Driven by ATP-hydrolysis, its central shaft rotates in 80 degrees and 40 degrees steps, interrupted by catalytic and ATP-waiting dwells. We recorded rotations and halts by means of microvideography in laboratory coordinates. A correlation with molecular coordinates was established by using an engineered pair of cysteines that, under oxidizing conditions, formed zero-length cross-links between the rotor and the stator in an orientation as found in crystals. The fixed orientation coincided with that of the catalytic dwell, whereas the ATP waiting dwell was displaced from it by +40 degrees . In crystals, the convex side of the cranked central shaft faces an empty nucleotide binding site, as if holding it open for arriving ATP. Functional studies suggest that three sites are occupied during a catalytic dwell. Our data imply that the convex side faces a nucleotide-occupied rather than an empty site. The enzyme conformation in crystals seems to differ from the conformation during either dwell of the active enzyme. A revision of current schemes of the mechanism is proposed.


Cavernous hemangioma of the conjunctiva: case report.
Cavernous hemangioma of the conjunctiva is rare and has been reported previously in only a few cases. In this report, the authors describe a 17-year-old boy with an isolated conjunctival cavernous hemangioma. The tumor was located in the caruncular region over the bulbar conjunctiva. The lesion had appeared as a red mark in early childhood, grown in the last 2 years, and was causing bloody tears occasionally. No associated vascular lesion was noted within the eye and orbit. After removal of the tumor, no recurrence or complication developed during the follow-up period.


Exploring the association between maternal mood and self-reports of sleep during the perinatal period.
To explore the psychological mechanisms involved in the close association between maternal mood and self-reports of sleep quality during the perinatal period using appraisal theory of emotions. Repeated measures. Antenatal clinics of a health center associated with the Northern Hospital in Melbourne, Australia. 122 pregnant women in their third trimester of gestation. Participants completed the Pittsburgh Sleep Quality Index (PSQI), the Edinburgh Postnatal Depression Scale, and an appraisal questionnaire during the third trimester of gestation, within 7 to 10 days after childbirth, and at 10 to 12 weeks postpartum. Correlational and regression analyses were used to explore the associations between sleep reports and appraisals. Self-reports of poor sleep quality, impaired daytime dysfunction due to poor sleep, and the global PSQI score were associated with a low perceived ability to cope practically and emotionally with motherhood-related issues as well as with negative expectations about the future. Appraisal dimensions associated with self-reports of poor sleep quality are similar to those related to maternal distress identified by previous research. This finding contributes to a better understanding of the association between self-reports of sleep and maternal mood. Practical implications are discussed.


DNA barcoding for identification of sand fly species (Diptera: Psychodidae) from leishmaniasis-endemic areas of Peru.
Phlebotomine sand flies are the only proven vectors of leishmaniases, a group of human and animal diseases. Accurate knowledge of sand fly species identification is essential in understanding the epidemiology of leishmaniasis and vector control in endemic areas. Classical identification of sand fly species based on morphological characteristics often remains difficult and requires taxonomic expertise. Here, we generated DNA barcodes of the cytochrome c oxidase subunit 1 (COI) gene using 159 adult specimens morphologically identified to be 19 species of sand flies, belonging to 6 subgenera/species groups circulating in Peru, including the vector species. Neighbor-joining (NJ) analysis based on Kimura 2-Parameter genetic distances formed non-overlapping clusters for all species. The levels of intraspecific genetic divergence ranged from 0 to 5.96%, whereas interspecific genetic divergence among different species ranged from 8.39 to 19.08%. The generated COI barcodes could discriminate between all the sand fly taxa. Besides its success in separating known species, we found that DNA barcoding is useful in revealing population differentiation and cryptic diversity, and thus promises to be a valuable tool for epidemiological studies of leishmaniasis.


Dynamic compositional changes of detergent-resistant plasma membrane microdomains during plant cold acclimation.
Plants increase their freezing tolerance upon exposure to low, non-freezing temperatures, which is known as cold acclimation. Cold acclimation results in a decrease in the proportion of sphingolipids in the plasma membrane in many plants including Arabidopsis thaliana. The decrease in sphingolipids has been considered to contribute to the increase in the cryostability of the plasma membrane through regulating membrane fluidity. Recently we have proposed a possibility of another important sphingolipid function associated with cold acclimation. In animal cells, it has been known that the plasma membrane contains microdomains due to the chanracteristics of sphingolipids and sterols, and the sphingolipid- and sterol-enriched microdomains are thought to function as platforms for cell signaling, membrane trafficking and pathogen response. In our research on characterization of microdomain-associated lipids and proteins in Arabidopsis, cold-acclimation-induced decrease in sphingolipids resulted in a decrease of microdomains in the plasma membrane and there were considerable changes in membrane transport-, cytoskeleton- and endocytosis-related proteins in the microdomains during cold acclimation. Based on these results, we discuss a functional relationship between the changes in microdomain components and plant cold acclimation.


Impact of critical limb ischemia on long-term cardiac mortality in diabetic patients undergoing percutaneous coronary revascularization.
Development of critical limb ischemia (CLI) has been reported as an independent predictor of cardiac mortality in diabetic patients. We aimed to determine whether CLI, managed in a structured setting of close collaboration between different vascular specialists and treated with early endovascular intervention, has any impact on long-term cardiac mortality of diabetic patients initially presenting with symptomatic coronary artery disease (CAD). We designed a prospective observational study of 764 consecutive diabetic patients undergoing percutaneous coronary intervention (PCI) in whom development of CLI was assessed by a dedicated diabetic foot clinic. Cardiac mortality at 4-year follow-up was the primary end point of the study. Among the 764 patients, 111 (14%) developed CLI (PCI-CLI group) and underwent revascularization of 145 limbs, with procedural success in 140 (96%). PCI-CLI patients at baseline had lower left ventricular ejection fraction (51 ± 11% vs. 53 ± 10%, P = 0.008), higher prevalence of dialysis (7% vs. 0.3%, P < 0.0001), and longer diabetes duration (13 ± 8 vs. 11 ± 7 years, P = 0.02) compared with PCI-only patients. At 4-year follow-up, cardiac mortality occurred in 10 (9%) PCI-CLI patients vs. 42 (6%) PCI-only patients (P = 0.2). Time-dependent Cox regression model for cardiac death revealed that CLI was not associated with an increased risk of cardiac mortality (hazard ratio 1.08 [95% CI 0.89-3.85]; P = 0.1). The development of promptly assessed and aggressively treated CLI was not significantly associated with increased risk of long-term cardiac mortality in diabetic patients initially presenting with symptomatic CAD.


Flow cytometric characterization of the DAOY medulloblastoma cell line for the cancer stem-like phenotype.
Side population (SP) analyses and CD133 expression have identified cells with stem-like potential in normal and cancerous tissue. Whether stem-like cells exist in cancer cell lines is hotly debated. We have interrogated the DAOY medulloblastoma cell line with respect to stem-like potential. Vital staining for Hoechst 33342 efflux capacity and CD133 immunophenotyping were performed on DAOY cells to assess the presence of the SP and the CD133 stem cell markers, respectively. SP/non-SP and CD133(+)/CD133(-) DAOY cells were sorted into separate fractions for limiting dilution analysis (tumor sphere assay) and asymmetric division assessment. SP/non-SP cells were also sorted separately for viability (XTT assay), cell size, cell cycle status, and proliferative capacity (carboxyfluorescein succinimidyl ester (CFSE)) evaluation. A minor proportion of cells displayed either the SP or the CD133(+) phenotypes. CD133 expression mapped to both the SP and non-SP compartments, with CD133(+) cells being enriched almost fourfold within the non-SP gate. The SP, non-SP, CD133(+), and CD133(-) fractions were all capable of reconstituting the original parental DAOY population. Slight clonogenic enrichment was observed in only the SP fraction; however, both CD133(+) and CD133(-) cells displayed equivalent stem cell-like frequencies. SP cells were resistant to Hoechst 33342-mediated toxicity relative to the parental population and differed from the non-SP cells with respect to increased cell size, decreased S-phase, and slightly decreased proliferative capacity. The multiparametric strategy described in this study revealed that the SP and CD133(+) subset may be two independent compartments. Our results highlight the need for new reliable specific cancer stem cell marker(s) as Hoechst 33342 efflux and CD133 expression might not be suitable for selectively isolating cancer stem-like cells from cell lines, as shown for the DAOY cells. As such, care must be used in interpreting therapeutic studies targeting the stem cell compartment of cancer cell lines.


Evaluation of plasma retinol-binding protein as a surrogate measure for plasma retinol concentrations.
Plasma retinol-binding protein (RBP) concentrations have been suggested as surrogate indicators for plasma retinol concentrations in the assessment of vitamin A status in less technologically developed settings. Plasma RBP was measured by radial immunodiffusion and plasma retinol by high performance liquid chromatography in a cross-sectional study of 900 pregnant women at the Queen Elizabeth Central Hospital, Blantyre, Malawi. The Spearman correlation coefficient between plasma RBP and retinol concentrations was 0.95 (p<0.0001). By linear regression, 0.70 micromol l(-1) retinol was equivalent to 21.1 mg l(-1) RBP. With these cut-off points for defining vitamin A deficiency, there was high concordance between categorical descriptions of deficiency using retinol and RBP by chi-square analysis (p<0.001). Measurement of plasma RBP by radial immunodiffusion is simple, inexpensive, and does not require expensive instrumentation. Plasma RBP concentrations measured by radial immunodiffusion are highly correlated with plasma retinol and can be used as a simple surrogate measure for vitamin A concentrations in large field studies.


Proteases produced by a proteolytic mutant of Clostridium botulinum type E.
A proteolytic mutant from Clostridium botulinum type E produced extracellular proteases after the end of exponential growth coinciding with the period of sporulation. Proteases were separated into four fractions by chromatography on a DEAE-cellulose column. One was a sulphydryl-dependent protease that also apparently required a divalent cation for enzyme activity since it was inhibited by EDTA. This enzyme hydrolysed synthetic amide and ester compounds containing an arginine residue, and showed some activity towards L-lysine methyl ester. It appeared that two of the other proteases were serine proteases and the fourth was a metal protease. These last three proteases did not require a thiol agent and did not hydrolyse any of the synthetic amides or esters examined. Only the sulphydryl-dependent protease could activate C. botulinum type B, E and F toxins. The ability of this enzyme to activate type B and E toxins was markedly lower than that of trypsin. The susceptibility of type B toxin to this protease was lower than that of type E toxin. C2 toxin was not activated by this enzyme. It is suggested that the sulphydryl-dependent protease in this proteolytic mutant of C. botulinum type E has properties similar to those of proteases from C. botulinum types B and F.


Targeted Chinese Medicine Delivery by A New Family of Biodegradable Pseudo-Protein Nanoparticles for Treating Triple-Negative Breast Cancer: In Vitro and In Vivo Study.
Triple negative breast cancer (TNBC) has the worst overall survival among all breast cancer subtypes; 80% of TNBC harbors TP53 mutation. Gambogic acid (GA) is an herbal compound isolated from the dry brownish gamboge resin of Garcinia hanburyi. A new family of biodegradable polymer, the folate (FA)-conjugated arginine-based poly(ester urea urethane)s nanoparticles (FA-Arg-PEUU NP), was developed as nano-carrier for GA. Its anti-TNBC effects and the underlying mechanism of action were examined. The average diameters of FA-Arg-PEUU NP and GA-loaded FA-Arg-PEUU NP (NP-GA) in water are around 165 and 220nm, respectively. Rhodamine-tagged FA-Arg-PEUU NP shows that the conjugation of FA onto Arg-PEUU NPs facilitates the internalization of FA-Arg-PEUU-NP into TNBC. Compared to free-GA at the same GA concentrations, NP-GA exhibits higher cytotoxicity in both TP53-mutated and non-TP53 expressed TNBC cells by increasing intrinsic and extrinsic apoptosis. In HCC1806-bearing xenograft mouse model, the targeted delivery of GA by the FA-Arg-PEUU-NP nano-carriers to the tumor sites results in a more potent anti-TNBC effect and lower toxicity towards normal tissues and organs when compared to free GA. Furthermore, NP-GA also reduces the tumor-associated macrophage (TAM) M1/M2 ratio, suggesting that the use of Arg-based nanoparticles as carriers for GA not only makes the surface of the nanoparticles positively charged, but also confers on to the nanoparticles an ability to modulate TAM polarization. Our data clearly demonstrate that NP-GA exhibits potent anti-TNBC effects with reduced off-target toxicity, which represents novel alternative targeted therapeutics for TNBC treatment.


Prevalence and Characteristics of Salmonella Serotypes Isolated from Fresh Produce Marketed in the United States.
Salmonella continues to rank as one of the most costly foodborne pathogens, and more illnesses are now associated with the consumption of fresh produce. The U.S. Department of Agriculture Microbiological Data Program (MDP) sampled select commodities of fresh fruit and vegetables and tested them for Salmonella, pathogenic Escherichia coli, and Listeria. The Salmonella strains isolated were further characterized by serotype, antimicrobial resistance, and pulsed-field gel electrophoresis profile. This article summarizes the Salmonella data collected by the MDP between 2002 and 2012. The results show that the rates of Salmonella prevalence ranged from absent to 0.34% in cilantro. A total of 152 isolates consisting of over 50 different serotypes were isolated from the various produce types, and the top five were Salmonella enterica serotype Cubana, S. enterica subspecies arizonae (subsp. IIIa) and diarizonae (subsp. IIIb), and S. enterica serotypes Newport, Javiana, and Infantis. Among these, Salmonella serotypes Newport and Javiana are also listed among the top five Salmonella serotypes that caused most foodborne outbreaks. Other serotypes that are frequent causes of infection, such as S. enterica serotypes Typhimurium and Enteritidis, were also found in fresh produce but were not prevalent. About 25% of the MDP samples were imported produce, including 65% of green onions, 44% of tomatoes, 42% of hot peppers, and 41% of cantaloupes. However, imported produce did not show higher numbers of Salmonella-positive samples, and in some products, like cilantro, all of the Salmonella isolates were from domestic samples. About 6.5% of the Salmonella isolates were resistant to the antimicrobial compounds tested, but no single commodity or serotype was found to be the most common carrier of resistant strains or of resistance. The pulsed-field gel electrophoresis profiles of the produce isolates showed similarities with Salmonella isolates from meat samples and from outbreaks, but there were also profile diversities among the strains within some serotypes, like Salmonella Newport.


Age-dependent Alteration in Mitochondrial Dynamics and Autophagy in Hippocampal Neuron of Cannabinoid CB1 Receptor-deficient Mice.
Endocannabinoid system activity contributes to the homeostatic defense against aging and thus may counteract the progression of brain aging. The cannabinoid type 1 (CB1) receptor activity declines with aging in the brain, which impairs neuronal network integrity and cognitive functions. However, the underlying mechanisms that link CB1 activity and memory decline remain unknown. Mitochondrial activity profoundly influences neuronal function, and age-dependent mitochondrial activity change is one of the known hallmarks of brain aging. As CB1 receptor is expressed on mitochondria and may regulate neuronal energy metabolism in hippocampus, we hypothesized that CB1 receptors might influence mitochondria in hippocampal neurons. Here, we found that CB1 receptor significantly affected mitochondrial autophagy (mitophagy) and morphology in an age-dependent manner. Serine 65-phosphorylated ubiquitin, a key marker for mitophagy, was reduced in adult CB1-deficient mice (CB1-KO) compared to those in wild type controls, particularly in CA1 pyramidal cell layer. Transmission electron microscopy (TEM) analysis showed reduced mitophagy-like events in hippocampus of adult CB1-KO. TEM analysis also showed that mitochondrial morphology in adult CB1-KO mice was altered shown by an increase in thin and elongated mitochondria in hippocampal neurons. 3D reconstruction of mitochondrial morphology after scanning electron microscopy additionally revealed an enhanced density of interconnected mitochondria. Altogether, these findings suggest that reduced CB1 signaling in CB1-KO mice leads to reduced mitophagy and abnormal mitochondrial morphology in hippocampal neurons during aging. These mitochondrial changes might be due to the impairments in mitochondrial quality control system, which links age-related decline in CB1 activity and impaired memory.


An artificial intelligence approach fit for tRNA gene studies in the era of big sequence data.
Unsupervised data mining capable of extracting a wide range of knowledge from big data without prior knowledge or particular models is a timely application in the era of big sequence data accumulation in genome research. By handling oligonucleotide compositions as high-dimensional data, we have previously modified the conventional self-organizing map (SOM) for genome informatics and established BLSOM, which can analyze more than ten million sequences simultaneously. Here, we develop BLSOM specialized for tRNA genes (tDNAs) that can cluster (self-organize) more than one million microbial tDNAs according to their cognate amino acid solely depending on tetra- and pentanucleotide compositions. This unsupervised clustering can reveal combinatorial oligonucleotide motifs that are responsible for the amino acid-dependent clustering, as well as other functionally and structurally important consensus motifs, which have been evolutionarily conserved. BLSOM is also useful for identifying tDNAs as phylogenetic markers for special phylotypes. When we constructed BLSOM with 'species-unknown' tDNAs from metagenomic sequences plus 'species-known' microbial tDNAs, a large portion of metagenomic tDNAs self-organized with species-known tDNAs, yielding information on microbial communities in environmental samples. BLSOM can also enhance accuracy in the tDNA database obtained from big sequence data. This unsupervised data mining should become important for studying numerous functionally unclear RNAs obtained from a wide range of organisms.


Enthalpy and interfacial free energy changes of water capillary condensed in mesoporous silica, MCM-41 and SBA-15.
The effect of confinement on the solid-liquid phase transitions of water was studied by using DSC and FT-IR measurements. Enthalpy changes upon melting of frozen water in MCM-41 and SBA-15 were determined as a function of pore size and found to decrease with decreasing pore size. The melting point also decreased almost monotonically with a decrease in pore size. Analysis of the Gibbs-Thomson relation on the basis of the thermodynamic data showed that there were two stages of interfacial free energy change after the constant region, i.e., below a pore size of 6.0 nm: a gradual decrease down to 3.4 nm and another decrease after a small jump upward. This fact demonstrates that the simple Gibbs-Thomson relation, i.e., a linear relation between the melting point change and the inverse pore size, is limited to the range not far from the melting point of bulk water. FT-IR measurements suggest that the decrease in enthalpy change and interfacial free energy change with decreasing pore size reflect the similarity of the structures of both liquid and solid phases of water in smaller pores at lower temperatures.


Feeding problems in Silver-Russell syndrome.
In order to identify the prevalence and severity of feeding problems in children with Silver-Russell syndrome (SRS) the feeding difficulties of 32 children with SRS and 32 age- and sex-matched control children were assessed using the Feeding Assessment Questionnaire (Harris and Booth 1992). Children's ages ranged from 2 to 11 years, with 19 male and 13 female pairs. In a subset of matched pairs, parent-child interaction at mealtimes was examined using video recording; food intake was also assessed over a 3-day period using weighed food diaries. Children with SRS experienced significantly more feeding problems than children without growth disorders. Common feeding problems for children with SRS were poor appetite, fussiness, slow feeding, and problems associated with oral-motor dysfunction. Feeding problems did not significantly affect intake of kilocalories, protein, fat, or carbohydrates. Mealtime interaction between children with SRS and their parents was significantly more negative than interactions between control-group children and their parents. The focus of intervention packages in SRS should be to reduce negative parent-child interactions and reduce parental anxiety about feeding, growth, and weight.


Effect of Strength Training on Lipid and Inflammatory Outcomes: Systematic Review With Meta-Analysis and Meta-Regression.
Background: The aim of this study was to perform a systematic review with meta-analysis and meta-regressions evaluating the effects of isolated strength training (ST), compared with a control group, on total cholesterol (TC), triglycerides (TG), low-density (LDL), high-density lipoprotein (HDL), C-reactive protein (CRP), and adiponectin of adults. Methods: Embase, PubMed, Cochrane, and Scopus data sources were searched up to May 2017. Clinical trials that compared ST with a control group of adults older than 18 years, which evaluated blood TC, TG, LDL, HDL, CRP, or adiponectin as an outcome were included. Random effect was used and the effect size (ES) was calculated by using the standardized mean difference with a 95% confidence interval. Results: ST promotes a reduction in TC (ES: -0.399; P < .001), TG (ES: -0.204; P = .002), LDL (ES: -0.451; P < .001), and CRP (ES: -0.542; P = .01) levels. In addition, ST is associated to an increase in HDL (ES: 0.363; P < .001) and adiponectin concentrations (ES: 1.105; P = .01). Conclusion: ST promotes decreases in TC, TG, LDL, and CRP levels and increases HDL and adiponectin concentrations. Thus, progressive ST could be a potential therapeutic option for improving abnormalities in lipid and inflammatory outcomes in adults.


Flavonoid-related basic helix-loop-helix regulators, NtAn1a and NtAn1b, of tobacco have originated from two ancestors and are functionally active.
The basic helix-loop-helix (bHLH) transcription factors (TFs) comprise one of the largest families of TFs involved in developmental and physiological processes in plants. Here, we describe the functional characterization of two bHLH TFs (NtAn1a and NtAn1b) isolated from tobacco (Nicotiana tabacum) flowers. NtAn1a and NtAn1b originate from two ancestors of tobacco, N. sylvestris and N. tomentosiformis, respectively. NtAn1a and NtAn1b share high sequence similarity with other known flavonoid-related bHLH TFs and are predominantly expressed in flowers. GUS expression driven by the NtAn1a promoter is consistent with NtAn1 transcript profile in tobacco flowers. Both NtAn1a and NtAn1b are transcriptional activators as demonstrated by transactivation assays using yeast cells and tobacco protoplasts. Ectopic expression of NtAn1a or NtAn1b enhances anthocyanin accumulation in tobacco flowers. In transgenic tobacco expressing NtAn1a or NtAn1b, both subsets of early and late flavonoid pathway genes were up-regulated. Yeast two-hybrid assays showed that NtAn1 proteins interact with the previously characterized R2R3-MYB TF, NtAn2. The NtAn1-NtAn2 complex activated the promoters of two key anthocyanin pathway genes, dihydroflavonol reductase and chalcone synthase. The promoter activation is severely repressed by dominant repressive forms of either NtAn1a or NtAn2, created by fusing the SRDX repressor domain to the TFs. Our results show that NtAn1 and NtAn2 act in concert to regulate the anthocyanin pathway in tobacco flowers and NtAn2 up-regulates NtAn1 gene expression.


The role of xanthine oxidase and the effects of antioxidants in ischemia reperfusion cell injury.
During last years considerable interest has been devoted to understand the role of oxygen radicals in the ischemia induced cell injury associated with reperfusion. In the brain and in others tissues, free radicals play a role as modulators of vascular tone as well as a cytotoxic role as part of the ischemia associated pathology. This review discusses methods for free radical detection in brain and in other tissues, mechanisms of radical production in the course of the ischemia reperfusion process, and the efficacy of potential antioxidant agents in post ischemia therapy, especially with respect to allopurinol, an inhibitor of xanthine oxidase, and the role of taurine and its derivatives as antioxidants in different organs including the brain.


[Effect of somatostatin on the myoelectrical activity of the stomach and small intestine in conscious dogs and rabbits].
In chronic experiments on dogs and rabbits, i.v. administration of somatostatin (3.3 to 330.0 ng/kg) exerted activating effects in resting and in hunger upon the myoelectrical activity of stomach, duodenum, small intestine and ileum whereas in the course of digestion the effects were inhibitory. Somatostatin exerted mainly an inhibitory effect on the myoelectrical activity of stomach and small intestine in rabbits. Possible mechanisms of the somatostatin effect upon the contractile activity of the stomach and duodenum smooth muscles are discussed.


The SLENDER gene of pea encodes a gibberellin 2-oxidase.
The amount of active gibberellin (GA) in plant tissues is determined in part by its rate of catabolism through oxidation at C-2. In pea (Pisum sativum L.) seeds, GA 2-oxidation is controlled by the SLN (SLENDER) gene, a mutation of which produces seedlings characterized by a slender or hyper-elongated phenotype. We cloned a GA 2-oxidase cDNA from immature pea seeds by screening an expression library for enzyme activity. The clone contained a full-length open reading frame encoding a protein of 327 amino acids. Lysate of bacterial cultures expressing the protein converted the C(19)-GAs, GA(1), GA(4), GA(9), and GA(20) to the corresponding 2beta-hydroxy products. GA(9) and GA(20) were also converted to GA(51) and GA(29) catabolites, respectively. The gene appeared to be one member of a small family of GA 2-oxidases in pea. Transcript was found predominantly in roots, flowers, young fruits, and testae of seeds. The corresponding transcript from sln pea contained a point mutation and did not produce active enzyme when expressed heterologously. RFLP analysis of a seedling population segregating for SLN and sln alleles showed the homozygous mutant allele co-segregating with the characteristic slender phenotype. We conclude that SLN encodes GA 2-oxidase.


Top-Down Cognitive and Linguistic Influences on the Suppression of Spontaneous Otoacoustic Emissions.
Auditory sensation is often thought of as a bottom-up process, yet the brain exerts top-down control to affect how and what we hear. We report the discovery that the magnitude of top-down influence varies across individuals as a result of differences in linguistic background and executive function. Participants were 32 normal-hearing individuals (23 female) varying in language background (11 English monolinguals, 10 Korean-English late bilinguals, and 11 Korean-English early bilinguals), as well as cognitive abilities (working memory, cognitive control). To assess efferent control over inner ear function, participants were presented with speech-sounds (e.g., /ba/, /pa/) in one ear while spontaneous otoacoustic emissions (SOAEs) were measured in the contralateral ear. SOAEs are associated with the amplification of sound in the cochlea, and can be used as an index of top-down efferent activity. Individuals with bilingual experience and those with better cognitive control experienced larger reductions in the amplitude of SOAEs in response to speech stimuli, likely as a result of greater efferent suppression of amplification in the cochlea. This suppression may aid in the critical task of speech perception by minimizing the disruptive effects of noise. In contrast, individuals with better working memory exert less control over the cochlea, possibly due to a greater capacity to process complex stimuli at later stages. These findings demonstrate that even peripheral mechanics of auditory perception are shaped by top-down cognitive and linguistic influences.


[Invasive pneumoccal disease in children in Oslo 1998-2004].
Invasive pneumococcal disease (IPD) is an important cause of morbidity and mortality in Norwegian children. This retrospective study included all children (under 16 years) with isolates of Streptococcus pneumoniae from a normally sterile site admitted to the Department of Paediatrics at Ullevaal University Hospital in the period 1998 to 2004. We studied the epidemiology, predisposing factors, clinical picture, antimicrobial resistance, outcome of IPD and the theoretical coverage of the 7-valent conjugate pneumococcal vaccine (PCV7) in these children. The isolates were tested for antimicrobial susceptibility, serogrouped and serotyped. 68 children were identified; 31 of them had one or more predisposing factors. Six children died, all of them had a predisposing factor. Six of the seven children who survived with sequelae were previously healthy. 67 of 68 isolates were fully susceptible to benzyl penicillin and 13 isolates showed intermediate susceptibility or resistance to erythromycin. Serogroups or serotypes were obtained in 66 children. 24 (36.8%) children fulfilled the criteria for PCV7. 35 (51.1%) children had serotypes covered by the vaccine. Only 12 (17.6%) fulfilled the criteria for PCV7 and had serotypes covered by it. Four of the six children who died had serotypes covered by PCV7. Invasive pneumococcal disease is a serious condition in children and vaccination can prevent disease in many children.


SPECT findings in mitochondrial encephalomyopathy.
We investigated the alterations in regional cerebral blood flow (rCBF) in mitochondrial encephalomyopathy (MEM), using [123I]N-isopropyl-p-iodoamphetamine (IMP) or 99mTc-hexamethyl propyleneamine oxime SPECT in 10 MEM patients. Four of the patients had MEM with lactic acidosis and strokelike episodes (MELAS), 2 had Kearns-Sayre syndrome (KSS), 1 had myoclonic epilepsy with ragged red fibers (MERRF) and 3 had cytochrome C oxidase deficiency (CCOD). Cerebral perfusion reserve was obtained from 6 patients (3 MELAS, 1 MERRF, 1 KSS, 1 CCOD) for a comparative analysis using the split-dose 123I-IMP SPECT method before and after the injection of acetazolamide. All patients with MELAS showed focal hypoperfusion in the parietal and/or occipital lobes. Follow-up studies (3 MELAS patients) revealed extension or improvement in the abnormal perfusion. The hypoperfused lesions were correlated with abnormal CT/MRI findings. Perfusion was normal in 1 MERRF, 2 KSS and 3 CCOD patients, whereas CT/MRI findings in 1 MERRF, 1 KSS and 1 CCOD patient were abnormal. The cerebral perfusion reserve in 3 MELAS patients was decreased significantly compared with that in patients with other types of MEM (MELAS 7.4%, other MEM 33.8%; p < 0.05). The rCBF was altered specifically in patients with MELAS, suggesting that brain perfusion SPECT will be useful in diagnosing and assessing such patients. The decreased cerebral perfusion reserve in patients with MELAS may represent an important feature of the pathogenesis of the strokelike episodes. The SPECT findings of patients with other types of MEM (MERRF, KSS and CCOD) were normal.


Responding to Natural and Industrial Disasters: Partnerships and Lessons Learned.
The aim of this study was to provide insights learned from disaster research response (DR2) efforts following Hurricane Harvey in 2017 to launch DR2 activities following the Intercontinental Terminals Company (ITC) fire in Deer Park, Texas, in 2019. A multidisciplinary group of academic, community, and government partners launched a myriad of DR2 activities. The DR2 response to Hurricane Harvey focused on enhancing environmental health literacy around clean-up efforts, measuring environmental contaminants in soil and water in impacted neighborhoods, and launching studies to evaluate the health impact of the disaster. The lessons learned after Harvey enabled rapid DR2 activities following the ITC fire, including air monitoring and administering surveys and in-depth interviews with affected residents. Embedding DR2 activities at academic institutions can enable rapid deployment of lessons learned from one disaster to enhance the response to subsequent disasters, even when those disasters are different. Our experience demonstrates the importance of academic institutions working with governmental and community partners to support timely disaster response efforts. Efforts enabled by such experience include providing health and safety training and consistent and reliable messaging, collecting time-sensitive and critical data in the wake of the event, and launching research to understand health impacts and improve resiliency.


Effectiveness of cognitive behavioural coaching in improving the well-being and retention of rural general practitioners.
To determine the relationship between cognitive behavioural coaching, the well-being of rural general practitioners (GPs), their intentions to leave and actual leaving of rural general practice. Quasi-experimental study utilising an intervention group of rural GPs and deidentified data for a baseline group and a control group consisting of the remaining population of rural GPs in South Australia. Rural general practice. Rural GPs in South Australia: intervention group (n = 69), baseline group (n = 205) and control group (n = 312). A 9-hour cognitive behavioural coaching program. Rural doctor distress, intention to leave rural general practice and actual retention rate of rural GPs. GPs who underwent cognitive behavioural coaching had lower rural doctor distress scores (mean = 24.50; confidence interval, 21.71-27.29) than the baseline group (mean = 28.63; confidence interval, 27.08-30.17). Before coaching, 81% of rural GPs in the intervention group had considered leaving general practice, which reduced to 40% after coaching (χ(2) (2) = 16.31, P < .001). Over a 3-year period, 94% of the coaching group remained in general practice compared with 80% of the control group (χ(2) (1) = 4.89, P = 0.027). Cognitive behavioural coaching reduced the stress levels of rural GPs who self-identified the need for managing stress and reduced their intention to leave rural general practice. Further, despite initially being more stressed compared with the general population of rural GPs, more GPs from the coaching group remained in rural general practice.


Modern Appraisal of Patency and Complications in Cerebral Bypass Surgery: A Single Institution Experience.
Cerebral bypass is a valuable surgical technique in well-selected patient populations. Updated clinical guidelines and improved surgical techniques warrant a contemporary reevaluation of the complications and patency to inform clinical practice and enhance postoperative patient care. To assess the complication rates and postoperative graft patency for the 3 most common indications for bypass surgery: moyamoya disease, intracranial atherosclerosis, and intracranial aneurysms. Perioperative notes of 175 consecutive bypass patients at a single institution were retrospectively identified to evaluate the clinical course and complications of surgery. The rate of total postoperative complications between moyamoya disease (9 of 98, 9.2%), intracranial atherosclerotic disease (7 of 57, 12.3%), and intracranial aneurysm (4 of 20, 20%) was not statistically different (P = .33). Immediate postoperative bypass patency was significantly higher in moyamoya disease (90 of 96, 93.8%) and intracranial atherosclerotic disease (48 of 51, 94.1%) than in intracranial aneurysm (13 of 18, 72.2%; P = .02). Intravenous heparin administration during bypass suturing was negatively associated with immediate postoperative patency (87% heparin patency vs 99% no heparin patency; P = .02). Double-barrel bypass trended toward an increased risk of wound healing complications (2 of 13, 15.4%) compared with the single-barrel bypass technique (4 of 156, 2.6%; P = .07). Cerebral bypass surgery remains an excellent surgical treatment for moyamoya disease, intracranial atherosclerosis, and intracranial aneurysms. This study suggests bypass is safer in moyamoya disease and intracranial atherosclerosis. Additional studies to clarify the risk of single-barrel vs double-barrel bypass and intraoperative heparin-stratified complications may be beneficial.


Two immediate-early pathogen-responsive members of the AtCMPG gene family in Arabidopsis thaliana and the W-box-containing elicitor-response element of AtCMPG1.
The Arabidopsis thaliana genome contains at least 50 predicted AtCMPG genes. The encoded protein family is defined by a common domain possessing four strictly conserved amino acid residues [Cys, Met, Pro, and Gly (CMPG)] that designate the family. Two members, AtCMPG1 and AtCMPG2, with high sequence similarity to the previously described, immediate-early pathogen-responsive PcCMPG1 gene from Petroselinum crispum were selected for analysis of their expression modes and defense-related promoter elements. Among the most striking similarities with PcCMPG1 were immediate-early transcriptional activation on infection or treatment with a pathogen-derived elicitor and the functional importance of a W-box-containing AtCMPG1 promoter element. Remarkably, this strongly pathogen/elicitor-responsive element, F, did not respond to wounding, in contrast to the AtCMPG1 promoter itself. Comparative analysis, both within the A. thaliana genome and across species, provided further insight into the large structural diversity of W-box-containing elements. Possible roles of AtCMPG proteins in regulatory processes are discussed with reference to a large variety of family members, partly with assigned functions, from plants as well as animals.


Reciprocal interaction between two cellular proteins, Puralpha and YB-1, modulates transcriptional activity of JCVCY in glial cells.
Cross communication between regulatory proteins is an important event in the control of eukaryotic gene transcription. Here we have examined the structural and functional interaction between two cellular regulatory proteins, YB-1 and Puralpha, on the 23-bp sequence element derived from the enhancer-promoter of the human polyomavirus JCV. YB-1 and Puralpha are single-stranded DNA binding proteins which recognize C/T- and GC/GA-rich sequences, respectively. Results from band shift studies demonstrated that while both proteins interact directly with their DNA target sequences within the 23-bp motif, each protein can regulate the association of the other one with the DNA. Affinity chromatography and coimmunoprecipitation provide evidence for a direct interaction between Puralpha and YB-1 in the absence of the DNA sequence. Ectopic expression of YB-1 and Puralpha in glial cells synergistically stimulated viral promoter activity via the 23-bp sequence element. Results from mutational studies revealed that residues between amino acids 75 and 203 of YB-1 and between amino acids 85 and 215 of Puralpha are important for the interaction between these two proteins. Functional studies with glial cells indicated that the region within Puralpha which mediates its association with YB-1 and binding to the 23-bp sequence is important for the observed activation of the JCV promoter by the Puralpha and YB-1 proteins. The results of this study suggest that the cooperative interaction between YB-1 and Puralpha mediates the synergistic activation of the human polyomavirus JCV genome by these cellular proteins. The importance of these findings for cellular and viral genes which are regulated by Puralpha and YB-1 is discussed.


Analysis of retinal vasculature using a multiresolution Hermite model.
This paper presents a vascular representation and segmentation algorithm based on a multiresolution Hermite model (MHM). A two-dimensional Hermite function intensity model is developed which models blood vessel profiles in a quad-tree structure over a range of spatial resolutions. The use of a multiresolution representation simplifies the image modeling and allows for a robust analysis by combining information across scales. Estimation over scale also reduces the overall computational complexity. As well as using MHM for vessel labelling, the local image modeling can accurately represent vessel directions, widths, amplitudes, and branch points which readily enable the global topology to be inferred. An expectation-maximization (EM) type of optimization scheme is used to estimate local model parameters and an information theoretic test is then applied to select the most appropriate scale/feature model for each region of the image. In the final stage, Bayesian stochastic inference is employed for linking the local features to obtain a description of the global vascular structure. After a detailed description and analysis of MHM, experimental results on two standard retinal databases are given that demonstrate its comparative performance. These show MHM to perform comparably with other retinal vessel labelling methods.


A new MRI rating scale for progressive supranuclear palsy and multiple system atrophy: validity and reliability.
To evaluate a standardised MRI acquisition protocol and a new image rating scale for disease severity in patients with progressive supranuclear palsy (PSP) and multiple systems atrophy (MSA) in a large multicentre study. The MRI protocol consisted of two-dimensional sagittal and axial T1, axial PD, and axial and coronal T2 weighted acquisitions. The 32 item ordinal scale evaluated abnormalities within the basal ganglia and posterior fossa, blind to diagnosis. Among 760 patients in the study population (PSP = 362, MSA = 398), 627 had per protocol images (PSP = 297, MSA = 330). Intra-rater (n = 60) and inter-rater (n = 555) reliability were assessed through Cohen's statistic, and scale structure through principal component analysis (PCA) (n = 441). Internal consistency and reliability were checked. Discriminant and predictive validity of extracted factors and total scores were tested for disease severity as per clinical diagnosis. Intra-rater and inter-rater reliability were acceptable for 25 (78%) of the items scored (≥ 0.41). PCA revealed four meaningful clusters of covarying parameters (factor (F) F1: brainstem and cerebellum; F2: midbrain; F3: putamen; F4: other basal ganglia) with good to excellent internal consistency (Cronbach α 0.75-0.93) and moderate to excellent reliability (intraclass coefficient: F1: 0.92; F2: 0.79; F3: 0.71; F4: 0.49). The total score significantly discriminated for disease severity or diagnosis; factorial scores differentially discriminated for disease severity according to diagnosis (PSP: F1-F2; MSA: F2-F3). The total score was significantly related to survival in PSP (p<0.0007) or MSA (p<0.0005), indicating good predictive validity. The scale is suitable for use in the context of multicentre studies and can reliably and consistently measure MRI abnormalities in PSP and MSA. Clinical Trial Registration Number The study protocol was filed in the open clinical trial registry (http://www.clinicaltrials.gov) with ID No NCT00211224.


Improved sensitivity of an enzyme immunoassay IDEIA for detecting Chlamydia trachomatis.
In tests on 375 genital tract specimens a commercially available enzyme immunoassay for Chlamydia trachomatis (IDEIA; Boots-Celltech) was found to have sensitivity values of 62% for men and 74% for women, and a specificity of 97% for both groups, relative to the results obtained by a fluorescence assay (Micro Trak; Syva). The positive predictive value and the negative predictive value of the immunoassay were 91% and 87%, respectively. Collection of samples for IDEIA in transport medium in plastic phials, as opposed to glass phials recommended by the manufacturer, had no effect on these values. Tests of the sensitivity of IDEIA using laboratory strains of C trachomatis showed that the assay detected chlamydial elementary bodies only at dilutions at least 10-fold lower than those at which they could be detected by Micro Trak. Tests of the specificity of the assay with microorganisms found in the genital tract, other than chlamydiae, showed that reactions occurred with a number of these. Testing three cervical swabs from the same patient, with the material taken into a single phial of transport medium, increased the sensitivity of IDEIA from 74% to 96%, without reducing the specificity which remained at 97%. It is concluded that this approach enchances the value of the test in a sexually transmitted disease clinic population and may do so in a population with a low prevalence of chlamydiae.


Measuring Self-Appraised Changes following Participation in an Intervention for Caregivers of Individuals with Dementia.
This study examined the psychometric properties of the Self-Appraisal of Change Scale, a measure designed to capture domains that are subjectively relevant to caregivers, associated with overall well-being, and amenable to change upon engaging in an intervention. Data from 107 caregivers of veterans with dementia enrolled in a randomized controlled trial of a telephone-delivered caregiver psychoeducation and support group program (the Telehealth Education Program (TEP)) were analyzed. The TEP, which was delivered by master's prepared social workers and a nurse dementia care manager, targeted caregivers' dementia-related knowledge, attitudes, and self-management skills. Analyses were conducted to determine the factor structure, reliability, validity, and measurement invariance of the scale. Factor analyses yielded a single-factor solution for the scale's nine items. The reliability and construct validity of the scale were also supported by the analyses. Multi-group analyses suggested that meaningful comparisons of scale items and total raw scores can be made across usual care (UC) and intervention groups. The findings offer preliminary support that the scale may represent a valid and reliable instrument that captures perceived changes in key domains (e.g. caregiving knowledge, attitudes, and skill) following participation in a psychoeducational and support intervention trial.


Novel archaeal adhesion pilins with a conserved N terminus.
Type IV pili play important roles in a wide array of processes, including surface adhesion and twitching motility. Although archaeal genomes encode a diverse set of type IV pilus subunits, the functions for most remain unknown. We have now characterized six Haloferax volcanii pilins, PilA[1-6], each containing an identical 30-amino-acid N-terminal hydrophobic motif that is part of a larger highly conserved domain of unknown function (Duf1628). Deletion mutants lacking up to five of the six pilin genes display no significant adhesion defects; however, H. volcanii lacking all six pilins (ΔpilA[1-6]) does not adhere to glass or plastic. Consistent with these results, the expression of any one of these pilins in trans is sufficient to produce functional pili in the ΔpilA[1-6] strain. PilA1His and PilA2His only partially rescue this phenotype, whereas ΔpilA[1-6] strains expressing PilA3His or PilA4His adhere even more strongly than the parental strain. Most surprisingly, expressing either PilA5His or PilA6His in the ΔpilA[1-6] strain results in microcolony formation. A hybrid protein in which the conserved N terminus of the mature PilA1His is replaced with the corresponding N domain of FlgA1 is processed by the prepilin peptidase, but it does not assemble functional pili, leading us to conclude that Duf1628 can be annotated as the N terminus of archaeal PilA adhesion pilins. Finally, the pilin prediction program, FlaFind, which was trained primarily on archaeal flagellin sequences, was successfully refined to more accurately predict pilins based on the in vivo verification of PilA[1-6].


Torulopsis bacarum, Torulopsis pustula and Torulopsis multis-gemmis sp. nov., three new yeasts from soft fruit.
Three new species of yeasts isolated from fresh soft fruit are described. Torulopsis pustula was isolated from blackcurrants, Torulopsis bacarum from blackcurrants, strawberries and raspberries, and Torulopsis multis-gemmis from raspberries. All differ from known species in the test substrates they utilize. In addition, cells of Torulopsis pustula often form a bulbous projection near their apices from which budding takes place. These buds are frequently pear- or heart-shaped.


Discovery of potent antiviral (HSV-1) quinazolinones and initial structure-activity relationship studies.
The discovery of antiviral activity of 2,3-disubstituted quinazolinones, prepared by a one-pot, three-component condensation of isatoic anhydride with amines and aldehydes, against Herpes Simplex Virus (HSV)-1 is reported. Sequential iterative synthesis/antiviral assessment allowed structure-activity relationship (SAR) generation revealing synergistic structural features required for potent anti-HSV-1 activity. The most potent derivatives show greater efficacy than acyclovir against acute HSV-1 infections in neurons and minimal toxicity to the host.


M.R.I diagnosis of tumours and tumour-like conditions affecting the pterygopalatine fossa.
To create awareness to the radiologist and clinicians for the magnetic resonance imaging (MRI) appearance of Pterygopalatine fossa (PTF) tumours and to evaluate the role of MRI in the diagnostic of PTF lesions. Retrospective evaluation of MRI features of 29 patients with pathologically proved pterygopalatine fossa (PTF) lesions was performed. The study included 18 males and 11 females with ages ranging between 15 and 68 years. All patients were examined on 1.5 Tesla magnets before and after injection of Gadolinium Meglumine. TI WI 5mm section sagittal scout views, followed by axial T1 5mm sections for the skull base and same sliced thickness during PD and T2 WI in axial and coronal planes. MRI features of various masses in pterygopalatine fossa were reviewed and correlated with those demonstrated in the literatures. Out of 29 PF masses, 7 masses were proved to be angiofibroma and 6 were nasopharyngeal carcinoma, entering the PTF. MRI is a useful imaging test which helps in the differential diagnosis of pterygopalatine fossa lesions with precise evaluation of their extensions and nature of lesion and helping to reach the correct diagnosis. However, MRI has limitation for evaluation of associated bony erosion, for which adjuvant CT scan is needed.


A practical approach to increasing intakes of n-3 polyunsaturated fatty acids: use of novel foods enriched with n-3 fats.
To assess the effects of providing a wide range of foodstuffs containing n-3 polyunsaturated fatty acids (PUFA), occurring naturally or from fortification, on intake and blood and tissue proportions of n-3 PUFA. Before/after dietary intervention study. Adelaide, Australia. 16 healthy males recruited from the community. Subjects were provided with a range of foodstuffs naturally containing n-3 PUFA (fresh fish, canned fish, flaxseed meal, canola oil) and items fortified with fish oil (margarine spread, milk, sausages, luncheon meat, french onion dip). Food choices were left to the discretion of each subject. Intake was estimated by diet diary. Blood was collected at-2, 0, 2, and 4 weeks for fatty acid analysis. Dietary intakes; plasma, platelet, and mononuclear cell phospholipid fatty acids. Consumption of n-3 PUFA increased significantly: alpha-linolenic acid (ALA) from 1.4 to 4.1 g/day (P<0.001), eicosapentaenoic acid (EPA) from 0.03 to 0.51 g/day (P<0.001), and docosahexaenoic acid (DHA) from 0.09 to 1.01 g/day (P<0.001). Linoleic acid (LA) intake decreased from 13.1 to 9.2 g/day (P<0.001). The proportions of EPA and DHA increased significantly in all phospholipid pools examined; plasma EPA from 1.13% of total fatty acids to 3.38% (P<0.001) and DHA from 3.76 to 7.23% (P<0.001); mononuclear cell EPA from 0.40 to 1.25% (P<0.001) and DHA from 2.33 to 4.08% (P<0.001); platelet EPA from 0.41 to 1.2% (P<0.001) and DHA from 1.64 to 3.07% (P<0.001). Incorporating fish oil into a range of novel commercial foods provides the opportunity for wider public consumption of n-3 PUFA with their associated health benefits. Dawes Scholarship, Royal Adelaide Hospital.


Urinary peptide analysis to predict the response to blood pressure medication.
The risk of Diabetic Kidney Disease (DKD) progression is significant despite renin-angiotensin system (RAS) blocking agents treatment. Current clinical tools cannot predict whether or not patients will respond to the treatment with RAS-inhibitors (RASi). We aimed to investigate if proteome analysis could identify urinary peptides as biomarkers that could predict the response to angiotensin-converting enzyme inhibitor (ACEi) and angiotensin receptor blockers (ARBs) treatment to avoid DKD progression. Furthermore, we investigated the comparability of the estimated glomerular filtration rate (eGFR), calculated using four different GFR-equations, for DKD progression. We evaluated urine samples from a discovery cohort of 199 diabetic patients treated with RASi. DKD progression was defined based on eGFR percentage slope results between visits (∼1 year) and for the entire period (∼3 year) based on the eGFR values of each GFR-equation. Urine samples were analysed using capillary electrophoresis coupled mass spectrometry. Statistical analysis was performed between the uncontrolled (patients who did not respond to RASi treatment) and controlled kidney function groups (patients who responded to the RASi treatment). Peptides were combined in a support vector machine-based model. The area under the receiver operating characteristic curve (AUC) was used to evaluate the risk prediction models in two independent validation cohorts treated with RASi. The classification of patients into uncontrolled and controlled kidney function varies depending on the GFR-equation used, despite the same sample set. We identified 227 peptides showing nominal significant difference and consistent fold changes between uncontrolled and controlled patients in at least three methods of eGFR calculation. These included fragments of collagens, alpha-1-antitrypsin, antithrombin-III, CD99 antigen, and uromodulin. A model based on 189 of 227 peptides (DKDp189) showed a significant prediction of non-response to the treatment/DKD progression in two independent cohorts. The DKDp189 model demonstrates potential as a predictive tool for guiding treatment with RASi in diabetic patients.


Attosecond Photoionization Dynamics: from Molecules over Clusters to the Liquid Phase.
Photoionization is a process taking place on attosecond time scales. How its properties evolve from isolated particles to the condensed phase is an open question of both fundamental and practical relevance. Here, we review recent work that has advanced the study of photoionization dynamics from atoms to molecules, clusters and the liquid phase. The first measurements of molecular photoionization delays have revealed the attosecond dynamics of electron emission from a molecular shape resonance and their sensitivity to the molecular potential. Using electron-ion coincidence spectroscopy these measurements have been extended from isolated molecules to clusters. A continuous increase of the delays with the water-cluster size has been observed up to a size of 4-5 molecules, followed by a saturation towards larger clusters. Comparison with calculations has revealed a correlation of the time delay with the spatial extension of the created electron hole. Using cylindrical liquid-microjet techniques, these measurements have also been extended to liquid water, revealing a delay relative to isolated water molecules that was very similar to the largest water clusters studied. Detailed modeling based on Monte-Carlo simulations confirmed that these delays are dominated by the contributions of the first two solvation shells, which agrees with the results of the cluster measurements. These combined results open the perspective of experimentally characterizing the delocalization of electronic wave functions in complex systems and studying their evolution on attosecond time scales.


Women and smoking: risks, impacts, and challenges.
Smoking among women has drawn increasing attention because of the increase (or less pronounced decrease) in its prevalence when compared with that observed for men, as well as because of the specific effects that smoking has on women's health. For the 2010 "World No Tobacco Day", the World Health Organization chose the theme "Gender and tobacco with an emphasis on marketing to women", with the aim of encouraging policies to combat marketing strategies employed by the tobacco industry and to curb the epidemic of smoking among women. In this article, we discuss the characteristics of smoking among women, addressing factors such as smoking prevalence, nicotine dependence, the role of the tobacco industry, health risks, approaches to smoking cessation, treatment strategies, and prevention measures.


Molecular phylogeny and genome evolution in the Drosophila virilis species group: duplications of the alcohol dehydrogenase gene.
Drosophila virilis is a prominent reference species for comparison with Drosophila melanogaster in regard to patterns and mechanisms of molecular and genomic evolution. Sequences were determined for 11 Adh genes from 8 species of the D. virilis species group, including species from both the virilis phylad and the montana subphylad. The genome of D. virilis contains a 6-kb duplication that includes the entire Adh coding region. The pattern of sequence identity within the duplication strongly suggests a recent gene-conversion event bordered by 36-bp indels. As in other Drosophila, the amino-acid coding region of Adh is encoded by three exons interrupted by two short introns. The promoter region includes 16 blocks of sequence that are well conserved in D. virilis, Drosophila hydei, and D. melanogaster. The developmental profile of Adh transcription suggests a distal/proximal promoter switch analogous to that in D. melanogaster. Duplicate Adh genes were also found in Drosophila montana and Drosophila lacicola, which apparently originated independently of that in D. virilis. The Adh genes in all species of the D. virilis group have among the lowest codon bias of any Adh genes so far reported in the genus Drosophila. Taking the low codon bias into account, we estimate the time of divergence between the virilis and montana clades as 9.0 +/- 0.7 Mya and the approximate time of divergence of D. virilis from other members of the virilis phylad as 2.6 +/- 0.4 Mya. The region of the D. virilis genome containing Adh, as well as the chromosome as a whole, gives evidence of extensive rearrangements relative to the genome of D. melanogaster.


Age-specific and sex-specific prevalence of cerebral β-amyloidosis, tauopathy, and neurodegeneration in cognitively unimpaired individuals aged 50-95 years: a cross-sectional study.
A new classification for biomarkers in Alzheimer's disease and cognitive ageing research is based on grouping the markers into three categories: amyloid deposition (A), tauopathy (T), and neurodegeneration or neuronal injury (N). Dichotomising these biomarkers as normal or abnormal results in eight possible profiles. We determined the clinical characteristics and prevalence of each ATN profile in cognitively unimpaired individuals aged 50 years and older. All participants were in the Mayo Clinic Study of Aging, a population-based study that uses a medical records linkage system to enumerate all individuals aged 50-89 years in Olmsted County, MN, USA. Potential participants are randomly selected, stratified by age and sex, and invited to participate in cognitive assessments; individuals without medical contraindications are invited to participate in brain imaging studies. Participants who were judged clinically as having no cognitive impairment and underwent multimodality imaging between Oct 11, 2006, and Oct 5, 2016, were included in the current study. Participants were classified as having normal (A-) or abnormal (A+) amyloid using amyloid PET, normal (T-) or abnormal (T+) tau using tau PET, and normal (N-) or abnormal (N+) neurodegeneration or neuronal injury using cortical thickness assessed by MRI. We used the cutoff points of standard uptake value ratio (SUVR) 1·42 (centiloid 19) for amyloid PET, 1·23 SUVR for tau PET, and 2·67 mm for MRI cortical thickness. Age-specific and sex-specific prevalences of the eight groups were determined using multinomial models combining data from 435 individuals with amyloid PET, tau PET, and MRI assessments, and 1113 individuals who underwent amyloid PET and MRI, but not tau PET imaging. The numbers of participants in each profile group were 165 A-T-N-, 35 A-T+N-, 63 A-T-N+, 19 A-T+N+, 44 A+T-N-, 25 A+T+N-, 35 A+T-N+, and 49 A+T+N+. Age differed by ATN group (p<0·0001), ranging from a median 58 years (IQR 55-64) in A-T-N- and 57 years (54-64) in A-T+N- to a median 80 years (75-84) in A+T-N+ and 79 years (73-87) in A+T+N+. The number of APOE ε4 carriers differed by ATN group (p=0·04), with carriers roughly twice as frequent in each A+ group versus the corresponding A- group. White matter hyperintensity volume (p<0·0001) and cognitive performance (p<0·0001) also differed by ATN group. Tau PET and neurodegeneration biomarkers were discordant in most individuals who would be categorised as stage 2 or 3 preclinical Alzheimer's disease (A+T+N-, A+T-N+, and A+T+N+; 86% at age 65 years and 51% at age 80 years) or with suspected non-Alzheimer's pathophysiology (A-T+N-, A-T-N+, and A-T+N+; 92% at age 65 years and 78% at age 80 years). From age 50 years, A-T-N- prevalence declined and A+T+N+ and A-T+N+ prevalence increased. In both men and women, A-T-N- was the most prevalent until age late 70s. After about age 80 years, A+T+N+ was most prevalent. By age 85 years, more than 90% of men and women had one or more biomarker abnormalities. Biomarkers of fibrillar tau deposition can be included with those of β-amyloidosis and neurodegeneration or neuronal injury to more fully characterise the heterogeneous pathological profiles in the population. Both amyloid- dependent and amyloid-independent pathological profiles can be identified in the cognitively unimpaired population. The prevalence of each ATN group changed substantially with age, with progression towards more biomarker abnormalities among individuals who remained cognitively unimpaired. National Institute on Aging (part of the US National Institutes of Health), the Alexander Family Professorship of Alzheimer's Disease Research, the Mayo Clinic, and the GHR Foundation.


Hookworms dynamically respond to loss of Type 2 immune pressure.
The impact of the host immune environment on parasite transcription and fitness is currently unknown. It is widely held that hookworm infections have an immunomodulatory impact on the host, but whether the converse is true remains unclear. Immunity against adult-stage hookworms is largely mediated by Type 2 immune responses driven by the transcription factor Signal Transducer and Activator of Transcription 6 (STAT6). This study investigated whether serial passage of the rodent hookworm Nippostrongylus brasiliensis in STAT6-deficient mice (STAT6 KO) caused changes in parasites over time. After adaptation to STAT6 KO hosts, N. brasiliensis increased their reproductive output, feeding capacity, energy content, and body size. Using an improved N. brasiliensis genome, we found that these physiological changes corresponded with a dramatic shift in the transcriptional landscape, including increased expression of gene pathways associated with egg production, but a decrease in genes encoding neuropeptides, proteases, SCP/TAPS proteins, and transthyretin-like proteins; the latter three categories have been repeatedly observed in hookworm excreted/secreted proteins (ESPs) implicated in immunosuppression. Although transcriptional changes started to appear in the first generation of passage in STAT6 KO hosts for both immature and mature adult stages, downregulation of the genes putatively involved in immunosuppression was only observed after multiple generations in this immunodeficient environment. When STAT6 KO-adapted N. brasiliensis were reintroduced to a naive WT host after up to 26 generations, this progressive change in host-adaptation corresponded to increased production of inflammatory cytokines by the WT host. Surprisingly, however, this single exposure of STAT6 KO-adapted N. brasiliensis to WT hosts resulted in worms that were morphologically and transcriptionally indistinguishable from WT-adapted parasites. This work uncovers remarkable plasticity in the ability of hookworms to adapt to their hosts, which may present a general feature of parasitic nematodes.


Association between suicidal behavior and impaired glucose metabolism in first-episode drug-naïve patients with schizophrenia.
Schizophrenia (SZ) patients have been reported to have comorbid suicidal behavior (SB) and impaired glucose metabolism in early psychosis, but it is unclear whether impaired glucose metabolism plays a role in the occurrence of SB in patients with first-episode drug-naïve (FEDN) SZ. Therefore, our main aim was to examine the relationship between SB and glucose metabolism in FEDN SZ patients. We recruited 319 FEDN SZ patients and collected information on their sociodemographic characteristics, clinical data, and glucose metabolism parameters. Participants' psychotic and depressive symptoms were assessed using the Positive and Negative Syndrome Scale (PANSS) and the Hamilton Depression Rating Scale (HAMD), respectively. Fasting plasma glucose and insulin levels were also measured. The percentage of FEDN SZ patients with SB was 45.5% (145/319). Compared to SZ patients without SB, SZ patients with SB exhibited higher scores on HAMD, PANSS positive subscale, as well as higher levels of fasting plasma glucose, fasting plasma insulin, and homeostasis model assessment of insulin resistance index (all p<0.05). Logistic regression analysis indicated that increased levels of insulin resistance (adjusted OR = 1.920), body mass index (adjusted OR = 0.931), and PANSS general psychopathology (adjusted OR = 1.041) were independently associated with SB. The Receiver Operating Characteristic Curve showed an Area Under Curve value of 0.732 for the combination of three factors in regression model to distinguish between SB and non-SB. Our results indicate that fasting glucose, fasting insulin, and insulin resistance are strongly associated with SB in FEDN SZ patients, suggesting that glucose metabolism abnormalities may be potential biomarkers of SB in SZ patients. Regular monitoring of glucose metabolism variables is essential for suicide prevention.


Determination of genotoxic potencies of pyrrolizidine alkaloids in HepaRG cells using the γH2AX assay.
Pyrrolizidine alkaloids (PAs) are secondary metabolites from plants that have been found in substantial amounts in herbal supplements, infusions and teas. Several PAs cause cancer in animal bioassays, mediated via a genotoxic mode of action, but for the majority of the PAs, carcinogenicity data are lacking. It is assumed in the risk assessment that all PAs have the same potency as riddelliine, which is considered to be one of the most potent carcinogenic PAs in rats. This may overestimate the risks, since many PAs are expected to have lower potencies. In this study we determined the concentration-dependent genotoxicity of 37 PAs representing different chemical classes using the γH2AX in cell western assay in HepaRG human liver cells. Based on these in vitro data, PAs were grouped into different potency classes. The group with the highest potency consists particularly of open diester PAs and cyclic diester PAs (including riddelliine). The group of the least potent or non-active PAs includes the monoester PAs, non-esterified necine bases, PA N-oxides, and the unsaturated PA trachelanthamine. This study reveals differences in in vitro genotoxic potencies of PAs, supporting that the assumption that all PAs have a similar potency as riddelliine is rather conservative.


Reduction of choline acetyltransferase activity in primary visual cortex in mild to moderate Alzheimer's disease.
Cholinergic deficits in the primary visual cortex (PVC) may underlie some of the abnormalities in visual processing and global cognitive performance in Alzheimer's disease (AD). To correlate measures of general cognition (Mini-Mental State Examination and Global Cognitive Score) and visuospatial function with choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) activities, and nerve growth factor protein levels in the PVC. The ChAT and AChE enzyme assays and a nerve growth factor protein enzyme-linked immunoabsorbent assay were performed on PVC tissue samples from subjects clinically diagnosed as having mild cognitive impairment (MCI), AD, or no cognitive impairment (NCI). Nuns, priests and brothers enrolled in the Religious Order Study, with annual premortem records of neuropsychological testing. Significant differences in ChAT activity, but not in AChE activity or nerve growth factor protein levels, were found among diagnostic groups (P = .049). The ChAT activity was lower in AD than in MCI or NCI (P<.01); MCI was not different from NCI. The PVC ChAT activity correlated with measures of overall cognitive function (Mini-Mental State Examination and Global Cognitive Score), but less so with a composite measure of visuospatial function. The reduction in ChAT activity in the PVC of mild to moderate AD, but not in MCI, might serve to distinguish between clinical and preclinical forms of the disease. It appears that this change relates to generalized cognitive abnormalities but not specifically to visuospatial function.


The direct connections of the C2 dorsal root ganglia in the Macaca irus monkey: relevance to the chiropractic profession.
In order to assess the direct connections of the C2 Dorsal Root Ganglion, the right ganglion in each of three Macaca Irus Monkeys was injected with labelled 3H-leucine. After survival times of 24 hours and autoradiographic techniques, the slides were read using light and dark field microscopes and mapped onto enlarged cross sectional drawings. Results show labelling into the lateral cervical nucleus, the central cervical nucleus and caudal to C5 of the spinal cord. Cephalad projections were into the medullary nuclei: cuneatus, lateral cuneatus, nucleus tractus solitarius, intercalatus, and X of the vestibular system. The relevance to chiropractic therapeutics is outlined, and this data will be used in a later paper to outline the office routine for assessing vertigo/dizziness of cervical origin.


Freeze-dried embedded specimens for biological microanalysis.
The main problems associated with freeze-drying of biological material for electron microscopy concern the freeze-drying temperatures and times necessary to minimize artifacts. Due to the many parameters involved these problems have to be resolved experimentally. It can be shown that good morphological preservation of chemically unfixed material is possible when freeze-drying is done exclusively in a temperature range between -80 degrees C and -50 degrees C. OsO4 vapour fixation of the freeze-dried tissue is not necessary and should be avoided because it may cause ion redistribution artifacts. Embedding at low temperature of properly freeze-dried material does not seem to disturb structure and ion distribution of the freeze-dried material. Hence, sections of such freeze-dried material and embedded biological material seem to be suitable for microanalysis. Preliminary micro-analytical results obtained from sections of freeze-dried and Lowicryl K11M embedded muscle reveal an uneven distribution of potassium in the sarcomeres similar to the visualized uneven distribution of the electron dense thallium (potassium surrogate) in frozen hydrated cryosections. A comparison of different cryomethods shows that freeze-drying and embedding is the simplest way to obtain stable thin sections of chemically unfixed biological material which, for instance, may be used for future microanalytical investigation of the interaction of proteins with physiological and non-physiological ions.


Crystal structure of the NADP+-dependent aldehyde dehydrogenase from Vibrio harveyi: structural implications for cofactor specificity and affinity.
Aldehyde dehydrogenase from the bioluminescent bacterium, Vibrio harveyi, catalyses the oxidation of long-chain aliphatic aldehydes to acids. The enzyme is unique compared with other forms of aldehyde dehydrogenase in that it exhibits a very high specificity and affinity for the cofactor NADP(+). Structural studies of this enzyme and comparisons with other forms of aldehyde dehydrogenase provide the basis for understanding the molecular features that dictate these unique properties and will enhance our understanding of the mechanism of catalysis for this class of enzyme. The X-ray structure of aldehyde dehydrogenase from V. harveyi has been solved to 2.5-A resolution as a partial complex with the cofactor NADP(+) and to 2. 1-A resolution as a fully bound 'holo' complex. The cofactor preference exhibited by different forms of the enzyme is predominantly determined by the electrostatic environment surrounding the 2'-hydroxy or the 2'-phosphate groups of the adenosine ribose moiety of NAD(+) or NADP(+), respectively. In the NADP(+)-dependent structures the presence of a threonine and a lysine contribute to the cofactor specificity. In the V. harveyi enzyme an arginine residue (Arg-210) contributes to the high cofactor affinity through a pi stacking interaction with the adenine ring system of the cofactor. Further differences between the V. harveyi enzyme and other aldehyde dehydrogenases are seen in the active site, in particular a histidine residue which is structurally conserved with phosphorylating glyceraldehyde-3-phosphate dehydrogenase. This may suggest an alternative mechanism for activation of the reactive cysteine residue for nucleophilic attack.


Activation of NF-κB via endosomal Toll-like receptor 7 (TLR7) or TLR9 suppresses murine herpesvirus 68 reactivation.
In order to understand and possibly treat B-cell malignancies associated with latent gammaherpesvirus infection, it is vital to understand the factors that control the balance between the two transcriptional states of gammaherpesviruses: latency and lytic replication. We used murine gammaherpesvirus 68 (MHV 68) as a model system to investigate how engagement of endosomal Toll-like receptors (TLRs) impacts reactivation from latency in vitro and establishment of latent infection in vivo. We found that treatment with TLR7 ligand R848 or TLR9 ligand CpG oligodeoxynucleotide (ODN) suppresses reactivation of MHV 68 in vitro. These suppressive effects correlated with the ability to activate cellular transcription factor NF-κB. Downregulation of TLR9 by RNA interference in vitro led to a reduction of nuclear levels of NF-κB p65 and consequently to an increase of spontaneous reactivation in cells latently infected with MHV 68, indicating that the TLR9 pathway suppresses spontaneous reactivation events. In vivo, sustained stimulation of TLR7 by repeated R848 treatment led to an increased frequency of infected splenocytes compared to mock-treated control results. Frequencies of infected splenic B cells in tlr7-/- or tlr9-/- mice after establishment of latency did not differ from those seen with their wild-type counterparts. Nevertheless, MHV 68-infected B cells from tlr9-/- mice showed a higher frequency of reactivation than B cells from wild-type or tlr7-/- mice in ex vivo reactivation assays. Thus, we show a suppressive effect of TLR7 or TLR9 triggering on MHV 68 reactivation that correlates with NF-κB activation and that the mere presence of a functional TLR9 signaling pathway contributes to dampen lytic gammaherpesvirus reactivation in infected cells. A hallmark of gammaherpesviruses is their establishment of latency in B cells that is reversible through lytic reactivation. Latency can result in B-cell malignancies. Activation of the innate immune system is thought to contribute to controlling the switch between the transcriptional states of latency and reactivation. Nevertheless, the mechanisms involved are not clear. Here, we show that engagement of Toll-like receptor 7 (TLR7) and TLR9 suppresses reactivation of murine gammaherpesvirus MHV 68 in vitro and that stimulation of TLR7 in vivo increases the frequency of infected cells. TLR7 and TLR9 are innate immunity sensors of nucleic acids localized in endosomes. Additionally, we demonstrate that impairment of TLR9 signaling in latently infected B cells leads to increased reactivation. Thus, activated endosomal TLR7 and TLR9 pathways play an important role in promoting establishment of latent gammaherpesvirus infection. Counteracting signaling of these pathways allows reactivation and could represent treatment targets in gammaherpesvirus-associated malignancies.


Human myeloma IgG half-molecules. Catabolism and biological properties.
A human IgG1 myeloma protein that has a delection in the third constant domain of the heavy chain (Cgamma3) and forms two-chain half-molecules was studied for its in vivo turnover and its ability to fix C1q and hemolytic complement, to bind to human lymphocytes, neutrophils, and monocytes, and to induce a passive cutaneous reaction in guinea pigs. In both man and monkeys, the half-molecule was rapidly catabolized and in part excreted into the urine. The half-life in man was 4.3 days and the fractional turnover 165% per day; 7.6% of the intravascular pool was excreted into the urine per day. Although the 7S four-chain myeloma protein could not be obtained in a pure form, the elimination from the serum of a partially purified preparation suggested that it was also rapidly catabolized. The unaggregated half-molecule neither formed complexes with C1q, cound to human lymphocytes, neutrophils, and monocytes, nor elicited a reverse passive cutaneous reaction in guinea pigs. In contrast, the aggregated half-molecule fixed hemolytic complement and bound to the human white cells similarly to an intact IgG1 myeloma protein. In order to explain the biological activities of this half-moleculr, it is postulated that IgG1 may have several (at least two) submolecular sites for a given biological activity that are localized on both the Cgamma2 and Cgamma3 domains. Proteins having both sites would be capable of binding to C1q and Fc cell receptors in unaggregated in order to obtain half-molecule, must be aggregated in order to obtain this binding.


Efficient calculation of van der Waals dispersion coefficients with time-dependent density functional theory in real time: application to polycyclic aromatic hydrocarbons.
The van der Waals dispersion coefficients of a set of polycyclic aromatic hydrocarbons, ranging in size from the single-cycle benzene to circumovalene (C(66)H(20)), are calculated with a real-time propagation approach to time-dependent density functional theory (TDDFT). In the nonretarded regime, the Casimir-Polder integral is employed to obtain C(6), once the dynamic polarizabilities have been computed at imaginary frequencies with TDDFT. On the other hand, the numerical coefficient that characterizes the fully retarded regime is obtained from the static polarizabilities. This ab initio strategy has favorable scaling with the size of the system--as demonstrated by the size of the reported molecules--and can be easily extended to obtain higher order van der Waals coefficients.


Effect of High-Voltage Atmospheric Cold Plasma Treatment on Germination and Heavy Metal Uptake by Soybeans (Glycine max).
The need to feed 9.9 billion people by 2050 will require the coordination of farming practices and water utilization by nutrient-dense plants and crops. High levels of lead (Pb), a toxic element that can accumulate in plants, can lead to toxicity in humans. With the development of novel treatment technologies, such as atmospheric cold plasma (ACP) and engineered nanoparticles (NPs), the time to germination and levels of heavy metals in food and feed commodities can be reduced. This study provides insight into the impact of plasma-activated water (PAW) on the germination rates and effects of soybean seeds, and the resultant combination effects of zinc oxide uptake in the presence of lead. Soybean seedlings were watered with PAW (treated for 3, 5, and 7 min at 30, 50, and 70 kV), and the germination and growth rate were monitored for 10 days. The germinated seedlings were then grown hydroponically in a nutrient solution, and the biomass of each example was measured. The PAW treatment that resulted in the best growth of soybean seeds was then exposed to Pb and zinc-oxide nanoparticles (ZnONPs) to investigate heavy metal uptake in the presence of nanoparticles. After acid digestion, the rate of heavy metal uptake by the soybean plants was evaluated using inductively coupled plasma-mass spectrometry. The PAW seeds grew and germinated more quickly, demonstrating that the plasma therapy had an effect. The rate of heavy metal uptake by the plants was also shown to be 5x lower in the presence of ZnONP.


Association between sclerostin and bone density in chronic spinal cord injury.
Spinal cord injury (SCI) results in profound bone loss due to muscle paralysis and the inability to ambulate. Sclerostin, a Wnt signaling pathway antagonist produced by osteocytes, is a potent inhibitor of bone formation. Short-term studies in rodent models have shown increased sclerostin in response to mechanical unloading that is reversed with reloading. These studies suggest that complete spinal cord injury, a condition resulting in mechanical unloading of the paralyzed lower extremities, will be associated with high sclerostin levels. We assessed the relationship between circulating sclerostin and bone density in 39 subjects with chronic SCI and 10 without SCI. We found that greater total limb bone mineral content was significantly associated with greater circulating levels of sclerostin. Sclerostin levels were reduced, not elevated, in subjects with SCI who use a wheelchair compared with those with SCI who walk regularly. Similarly, sclerostin levels were lower in subjects with SCI who use a wheelchair compared with persons without SCI who walk regularly. These findings suggest that circulating sclerostin is a biomarker of osteoporosis severity, not a mediator of ongoing bone loss, in long-term, chronic paraplegia. This is in contrast to the acute sclerostin-mediated bone loss shown in animal models of mechanical unloading in which high sclerostin levels suppress bone formation. Because these data indicate important differences in the relationship between mechanical unloading, sclerostin, and bone in chronic SCI compared with short-term rodent models, it is likely that sclerostin is not a good therapeutic target to treat chronic SCI-induced osteoporosis.


Stereotactic mesencephalotomy for cancer - related facial pain.
Cancer-related facial pain refractory to pharmacologic management or nondestructive means is a major indication for destructive pain surgery. Stereotactic mesencephalotomy can be a valuable procedure in the management of cancer pain involving the upper extremities or the face, with the assistance of magnetic resonance imaging (MRI) and electrophysiologic mapping. A 72-year-old man presented with a 3-year history of intractable left-sided facial pain. When pharmacologic and nondestructive measures failed to provide pain alleviation, he was reexamined and diagnosed with inoperable hard palate cancer with intracranial extension. During the concurrent chemoradiation treatment, his cancer-related facial pain was aggravated and became medically intractable. After careful consideration, MRI-based stereotactic mesencephalotomy was performed at a point 5 mm behind the posterior commissure, 6 mm lateral to and 5 mm below the intercommissural plane using a 2-mm electrode, with the temperature of the electrode raised to 80℃ for 60 seconds. Up until now, the pain has been relatively well-controlled by intermittent intraventricular morphine injection and oral opioids, with the pain level remaining at visual analogue scale 4 or 5. Stereotactic mesencephalotomy with the use of high-resolution MRI and electrophysiologic localization is a valuable procedure in patients with cancer-related facial pain.


A simple method for the determination of lipid composition of human bile.
The Entero-Test, a device for easy sampling of gastrointestinal contents, including bile, has been used for determination of biliary lipid composition. The device consists of a weighted gelatin capsule containing 140 cm of a highly absorbent nylon line. The capsule is swallowed while one end of the string is taped to the face. After 3.5 h, when the line has reached the duodenum, gallbladder contraction is stimulated by intramuscular administration of ceruletide. The line is pulled out, and the last 15 cm are eluted four times in methanol. Total bile acids (by 3 alpha-hydroxysteroid-dehydrogenase assay), individual bile acids (by high performance liquid chromatography), phospholipids (by assay of lipid-soluble phosphorus), and cholesterol (by gas-liquid chromatography) are determined in the eluate. Tests in vitro demonstrated no preferential binding and a good recovery of biliary lipids from the thread. Similar values of biliary cholesterol saturation were obtained by means of duodenal intubation and of the Entero-Test in a series of 12 subjects (r = 0.952). In 5 subjects, individual bile acids were also measured and were found to be similar with both techniques (r = 0.948). When the test was repeated over 3 days in a series of 7 subjects, biliary cholesterol saturation was found to be remarkably reproducible (CV = 7.6%). Thus, the Entero-Test is a convenient technique for the determination of biliary lipid composition, which can be particularly useful in longitudinal studies.


Ca2+ regulation of tight-junction permeability and structure in Necturus gallbladder.
To explore the role of Ca2+ in tight-junction permeability, the Necturus gallbladder was exposed to varying Ca2+ concentrations and to the Ca2+ ionophore A23187 added to the mucosal side (1.9 X 10(-6) to 6.8 X 10(-5) M). Electrophysiological parameters measured in an Ussing-type chamber were correlated with tight-junction morphology revealed by freeze-fracture electron microscopy. In Ca2+-free bathing media, transepithelial resistance decreases and tight-junctional ultrastructure is fragmented. In 1.8 mM Ca2+ media, A23187 induces an initial drop in transepithelial resistance, followed by an increase in transepithelial resistance to a value 20% above base line. At peak response to A23187, NaCl diffusion potentials decrease. Freeze-fracture replicas reveal that the number of junctional strands increase pari passu with junctional depth. Both physiological and morphological changes were partially reversible. The initial decrease in transepithelial resistance coincided with a persistent hyperpolarization of the mucosal cell membrane potential difference and a decrease in the mucosal-to-serosal cell membrane resistance ratio. Thus A23187 alters both the transcellular and paracellular pathway, resulting in opposing effects on transepithelial resistance.


Feasibility of A Novel On-Site Detection Method for Aflatoxin in Maize Flour from Markets and Selected Households in Kampala, Uganda.
In sub-Saharan Africa, there is a high demand for affordable and accessible methods for on-site detection of aflatoxins for appropriate food safety management. In this study, we validated an electrochemical immunosensor device by the on-site detection of 60 maize flour samples from six markets and 72 samples from households in Kampala. The immunosensor was successfully validated with a linear range from 0.7 ± 0.1 to 11 ± 0.3 µg/kg and limit of detection (LOD) of 0.7 µg/kg. The maize flour samples from the markets had a mean total aflatoxin concentration of 7.6 ± 2.3 µg/kg with approximately 20% of the samples higher than 10 µg/kg, which is the maximum acceptable level in East Africa. Further down the distribution chain, at the household level, approximately 45% of the total number contained total aflatoxin levels higher than the acceptable limit. The on-site detection method correlated well with the established laboratory-based HPLC and ELISA-detection methods for aflatoxin B₁ with the correlation coefficients of 0.94 and 0.98, respectively. This study shows the feasibility of a novel on-site detection method and articulates the severity of aflatoxin contamination in Uganda.