[Development and Validation of the New Version of Spirituality Assessment Scale].
The purpose of this study was to develop a new version of Spirituality Assessment Scale (N-SAS) and verify its reliability and validity. The total of 59 preliminary items for the N-SAS were selected through a literature review, two rounds of experts' content validation, cognitive interviews, and pre-tests. Verification of its reliability and validity was divided into two phases. In Phase I, questionnaires were collected from 219 adults. Reliability was tested using Cronbach's alpha, validity with item analysis, and exploratory factor analysis. In Phase II, questionnaires developed based on the results of Phase I were collected from 225 adults. Reliability was tested using Cronbach's alpha, validity with confirmatory factor analysis, and criterion validity. The final version of the N-SAS comprised two dimensions (vertical and horizontal), four domains (relationship with God; meaning of life and self-integration; self-transcendence; and relationship with others, neighborhoods, and nature), and 44 items were identified. Total Cronbach's α was .97; those of each subscale ranged from .79 to .98. N-SAS scores were positively correlated with the scores of Howden's Spiritual Assessment Scale (r=.81, p<.001). Findings suggest that the N-SAS can be used to measure spirituality in adults. The use of N-SAS is expected to facilitate perceiving patient's spiritual needs and providing spiritual care.


Inhibition of mitogen activated protein kinase activity induces parthenogenetic activation and increases cyclin B accumulation during porcine oocyte maturation.
The inhibition of mitogen activated protein kinase (MAPK) activation during porcine oocyte maturation leads to decreased maturation promoting factor (MPF) activity and to the induction of parthenogenetic activation. In the present study, in order to analyze the mechanism underlying the suppression of MPF activity in MAPK-inhibited porcine oocytes, we injected mRNA of SASA-MEK, a dominant negative MAPK kinase, or antisense RNA of c-mos, a MAPK kinase kinase, into immature porcine oocyte cytoplasm. The injection of SASA-MEK mRNA or c-mos antisense RNA inhibited the MAPK activity partially or completely, respectively, decreased the MPF activity slightly or significantly, respectively, and induced parthenogenetic activation in 17.1% or 96.6% of mature oocytes, respectively, although no parthenogenetic activation was observed in the control oocytes. Immunoblotting experiments revealed that cyclin B accumulation in these MAPK-suppressed porcine oocytes was increased significantly after 50 h of culture and that a considerable amount of MPF was converted into inactive pre-MPF by hyperphosphorylation. These results indicate that the inhibition of MAPK activity in porcine oocytes did not promote cyclin B degradation but rather suppressed it; also the decrease in MPF activity in MAPK-suppressed porcine oocytes correlated with the conversion of active MPF into inactive pre-MPF.


The possible involvement of genetic variants of NET1 in the etiology of attention-deficit/hyperactivity disorder comorbid with oppositional defiant disorder.
Attention-deficit/hyperactivity disorder (ADHD) and oppositional defiant disorder (ODD) often coexist and shared some genetic influences. Evidence from the existing literature indicated that comorbid with ODD may increase the heterogeneity of ADHD genetics. Our present study sought to investigate the role of norepinephrine transporter gene (NET1) for ADHD comorbid with ODD. Six single nucleotide polymorphisms (SNPs) of NET1 were genotyped for a total of 1,815 ADHD cases, including 587 subjects (32.3%) with ODD. Chi-square tests were conducted for pseudo case-control study comparing allelic and genotypic distributions between ADHD with and without ODD. Among them, there were 1,249 probands together with their parents composing trios for family-based association studies using transmission disequilibrium tests (TDTs). In addition, 1,337 ADHD probands have detailed information of ODD symptoms and were included for quantitative analyses with genotypes using analyses of covariance (ANCOVA). To consider the overlap and correlation of other comorbidities with ODD and eliminate their potential confounding effect, we further repeated above analyses for 'pure ADHD+ODD' versus 'ADHD-only' after excluding other comorbidities except for ODD. The pseudo case-control study showed different allelic and genotypic distributions of SNP rs3785143 between ADHD with ODD and those without ODD. Family-based association tests indicated overtransmission of the T allele of rs3785143 in ADHD with ODD trios, but no biased transmission in those without ODD. ANCOVA showed association between genotypes of rs3785143 with ODD symptoms in ADHD probands, especially with 'Argumentative/Defiant Behavior (ADB)' dimension after controlling gender, age, clinical subtypes and intelligence. Above association still existed after removing the samples with other comorbidities. NET1 was associated with comorbidity of ODD and ODD symptoms in ADHD probands. Our findings emphasize the importance of considering the comorbidity of ODD in ADHD genetic studies, especially ADHD with ADB. However, further replication in independent sample or different populations is still needed.


Submucosal tunneling endoscopic resection for upper gastrointestinal multiple submucosal tumors originating from the muscular propria layer: a feasibility study.
In recent years, submucosal tunneling endoscopic resection (STER) was applied more and more often for single gastrointestinal (GI) submucosal tumor (SMT). However, little is known about this technique for treating multiple SMTs in GI tract. In the present study, we investigated the feasibility and outcome of STER for upper GI multiple SMTs originating from the muscularis propria (MP) layer. A feasibility study was carried out including a consecutive cohort of 23 patients with multiple SMTs from MP layer in esophagus, cardia, and upper corpus who were treated by STER from June 2011 to June 2014. Clinicopathological, demographic, and endoscopic data were collected and analyzed. All of the 49 SMTs were resected completely by STER technique. Furthermore, only one tunnel was built for multiple SMTs of each patient in this study. En bloc resection was achieved in all 49 tumors. The median size of all the resected tumors was 1.5 cm (range 0.8-3.5 cm). The pathological results showed that all the tumors were leiomyoma, and the margins of the resected specimens were negative. The median procedure time was 40 min (range: 20-75 min). Gas-related complications were of the main complications, the rates of subcutaneous emphysema and pneumomediastinum, pneumothorax, and pneumoperitoneum were 13.0%, 8.7% and 4.3%. Another common complication was thoracic effusion that occurred in 2 cases (8.7%), among which only 1 case (4.3%) with low-grade fever got the drainage. Delayed bleeding, esophageal fistula or hematocele, and infection in tunnel were not detected after the operation there were no treatment-related deaths. The median hospital stay was 4 days (range, 2-9 days). No residual or recurrent lesion was found during the follow-up period (median 18, ranging 3-36 months). Submucosal tunneling endoscopic resection is a safe and efficient technique for treating multiple esophageal SMTs originating from MP layer, which can avoid patients suffering repeated resections.


Linear models in color constancy theory.
Linear models in color constancy theory face, at least, five major problems different in nature, to be solved as a prerequisite for a satisfactory theory of the phenomenon. Solutions are proposed.


Ultrasound-guided injection of the cranial tibial artery for stem cell administration in horses.
A technique for intra-arterial injection of mesenchymal stem cells (MSC) has been established for front limbs with the use of the median artery. This approach has been proposed for treatment of soft tissue injuries of the equine distal limb. A technique has not been validated yet for hindlimb injection. To assess the feasibility of injection of the cranial tibial artery in horses, and to evaluate the distribution and persistence of MSC after injection. In vivo experiment. In a first phase, the cranial tibial arteries of both hindlimbs of three research horses were catheterised with ultrasound guidance under general anaesthesia and injected with iodinated contrast. In the second phase, iodinated contrast was injected in three standing sedated horses with ultrasound guidance. In the final phase, 99m Technetium-HMPAO labelled allogenic bone marrow derived equine MSC were injected under standing sedation with the same technique in three other horses. Scintigraphy was used to assess MSC distribution and persistence for 24 h. Ultrasound was performed 24 h after injection to assess vessel impairment. Arterial injection was achieved in all 18 limbs without any significant complications. Mild partial periarterial injection was observed in four limbs. Scintigraphic images demonstrated diffuse MSC distribution from the tarsal area to the foot. Persistence decreased over time but signal was still present at 24 h. Limited retention of the radiolabel in the MSC. Ultrasound-guided injection of the cranial tibial artery can be performed both under general anaesthesia and standing sedation in horses. This technique could be used for MSC treatment of equine proximal suspensory desmopathy or other injuries in the distal hindlimb.


UV irradiation of lymphocytes triggers an increase in intracellular Ca2+ and prevents lectin-stimulated Ca2+ mobilization: evidence for UV- and nifedipine-sensitive Ca2+ channels.
UV irradiation induces in vitro and in vivo immunosuppression. Because mobilization of intracellular calcium ([Ca2+]i) represents a central step in cell activation and immune response, we investigated the effect of UV irradiation on Ca2+ homeostasis. Using indo-1 and cytofluorometry, [Ca2+]i kinetics in UVC- or UVB-exposed human peripheral blood leukocytes (PBL) and Jurkat cells were determined in parallel with functional assays. Increases in [Ca2+]i were observed within 2-3 h of irradiation; these increases were UV-dose dependent and reached maxima of 240% and 180% above baseline level (130 nM) for UVB and UVC, respectively. The UV-induced [Ca2+]i rise was predominantly due to influx of extracellular calcium, and it was more pronounced in T than in non-T cells. Concurrent with [Ca2+]i shifts following UV treatment, there was a loss of ability to respond to phytohemagglutinin (PHA) or to proliferate or stimulate in mixed leukocyte culture. This loss of function appeared to be related not only to UV-induced calcium shifts, but also to effects of UV irradiation on the plasma membrane. No [Ca2+]i mobilization was induced by gamma irradiation, and gamma-irradiated cells showed a normal [Ca2+]i increase in response to PHA. UV-induced Ca2+ flux into the cells was blocked by nifedipine. These data indicate that UV and gamma irradiation have different effects on lymphocyte membranes and suggest that a disruption of Ca2+ homeostasis may be involved in UV-induced lymphocyte inhibition. The data suggest, furthermore, the presence of Ca2+ channels in lymphocyte membranes that are sensitive to UV irradiation and Ca2+ channel blockers such as nifedipine.


Effects of NH4CL application and removal on astrocytes and endothelial cells.
Hepatic encephalopathy (HE) is a complex disorder associated with increased ammonia levels in the brain. Although astrocytes are believed to be the principal cells affected in hyperammonemia (HA), endothelial cells (ECs) may also play an important role by contributing to the vasogenic effect of HA. Following acute application and removal of NH4Cl on astrocytes and endothelial cells, we analyzed pH changes, using fluorescence imaging with BCECF/AM, and changes in intracellular Ca2+ concentration ([Ca2+]i), employing fluorescence imaging with Fura-2/AM. Using confocal microscopy, changes in cell volume were observed accompanied by changes of [Ca2+]i in astrocytes and ECs. Exposure of astrocytes and ECs to 1 - 20 mM NH4Cl resulted in rapid concentration-dependent alkalinization of cytoplasm followed by slow recovery. Removal of the NH4Cl led to rapid concentration-dependent acidification, again followed by slow recovery. Following the application of NH4Cl, a transient, concentration-dependent rise in [Ca2+]i in astrocytes was observed. This was due to the release of Ca2+ from intracellular stores, since the response was abolished by emptying intracellular stores with thapsigargin and ATP, and was still present in the Ca2+-free bathing solution. The removal of NH4Cl also led to a transient concentration-dependent rise in [Ca2+]i that resulted from Ca2+ release from cytoplasmic proteins, since removing Ca2+ from the bathing solution and emptying intracellular Ca2+ stores did not eliminate the rise. Similar results were obtained from experiments on ECs. Following acute application and removal of NH4Cl no significant changes in astrocyte volume were detected; however, an increase of EC volume was observed after the administration of NH4Cl, and EC shrinkage was demonstrated after the acute removal of NH4Cl. This study reveals new data which may give a more complete insight into the mechanism of development and treatment of HE.


Homodyne interferometry with quadrature fringe detection for absolute gravimeter.
An absolute gravimeter is based on interferometric displacement measurement during repeated free falls of a target mirror in a high vacuum. A suitable homodyne quadrature laser interferometer is developed and reported. Standard deviation of the mean absolute gravity value in a typical two-day measurement session is less than 1×10⁻⁸ m/s². Additionally, digital demodulation of the interference signal using a 90° phase-shift filter based on the Hilbert transform is presented. A combination of optical and digital quadrature phase shifts is shown to be helpful to improve accuracy in homodyne interferometers with the accelerated target mirror.


Management of Patients With Hepatitis C Virus, Monoclonal Gammopathy of Undetermined Significance, and Multiple Myeloma.
Background and Aim: The vast majority of the 2.7 million individuals in the United States who are currently infected with hepatitis C virus (HCV) were born between 1945 and 1965. The median age of these patients in this particular generation at the time of this writing was 55 years. In the general population, older age is a risk factor for multiple myeloma (MM) and other monogammopathies. As the baby boomer population ages, HCV providers are increasingly likely to encounter HCV-infected patients with a monoclonal gammopathy. Guidelines for managing these patients are needed. Methods: We conducted a detailed case series investigation of 4 HCV-positive patients with MM or a monoclonal gammopathy disorder. Patients were followed at the Mount Sinai Faculty Practice liver clinic. We also performed a detailed review of the literature exploring if there is any known association between HCV, MM, and monoclonal gammopathy. Results and Conclusions: There is no convincing evidence of a causal association between HCV and MM. HCV is linked to type II and type III cryoglobulinemia, specific kinds of monoclonal gammopathies of determinable significance. Whether a link exists between HCV and MM or monoclonal gammopathy of undetermined significance is unclear. Our case series provides the first evidence that modern HCV treatments with direct-acting antivirals can be safely and effectively co-administered with MM chemotherapy.


UNiquant, a program for quantitative proteomics analysis using stable isotope labeling.
Stable isotope labeling (SIL) methods coupled with nanoscale liquid chromatography and high resolution tandem mass spectrometry are increasingly useful for elucidation of the proteome-wide differences between multiple biological samples. Development of more effective programs for the sensitive identification of peptide pairs and accurate measurement of the relative peptide/protein abundance are essential for quantitative proteomic analysis. We developed and evaluated the performance of a new program, termed UNiquant, for analyzing quantitative proteomics data using stable isotope labeling. UNiquant was compared with two other programs, MaxQuant and Mascot Distiller, using SILAC-labeled complex proteome mixtures having either known or unknown heavy/light ratios. For the SILAC-labeled Jeko-1 cell proteome digests with known heavy/light ratios (H/L = 1:1, 1:5, and 1:10), UNiquant quantified a similar number of peptide pairs as MaxQuant for the H/L = 1:1 and 1:5 mixtures. In addition, UNiquant quantified significantly more peptides than MaxQuant and Mascot Distiller in the H/L = 1:10 mixtures. UNiquant accurately measured relative peptide/protein abundance without the need for postmeasurement normalization of peptide ratios, which is required by the other programs.


Preoperative CYFRA 21-1 and CEA as prognostic factors in patients with stage I non-small cell lung cancer: external validation of a prognostic score.
To validate the prognostic value of preoperative levels of CYFRA 21-1, CEA and the corresponding tumor marker index (TMI) in patients with stage I non-small cell lung cancer (NSCLC). Two hundred forty stage I NSCLC patients (80 in pT1 and 160 in pT2; 100 squamous cell carcinomas, 91 adenocarcinomas, 32 large-cell carcinomas, 17 with other histologies; 171 males and 69 females) who had complete resection (R0) between 1986 and 2004 were included in the analysis. CYFRA 21-1 and CEA were measured using the Elecsys system (Roche) and AxSym-System (Abbott), respectively. Univariate analysis was performed using the Kaplan-Meier method to identify potential associations between survival and age, gender, CYFRA 21-1, CEA and TMI. Overall 3- and 5-year survival rates were 74 and 64%, respectively. Male gender (p = 0.0009) and age >70 years (p = 0.0041) were associated with a worse prognosis; there were no differences between pT1 and pT2 nor between histological subtypes. Three-year survival was 72% for CYFRA 21-1 levels >3.3 ng/ml versus 75% for levels <or=3.3 ng/ml, 71% for CEA > 6.7 ng/ml versus 75% for CEA <or=6.7 ng/ml (both p values >0.05). Corresponding 5-year survival rates were near 64% both for patients with CYFRA 21-1 values above and below the cutoff (3.3 ng/ml), and 49 and 66% for patients with values above and below the CEA cutoff (6.7 ng/ml), respectively (both p values >0.05). Overall survival did not vary in the different TMI risk groups (p = 0.73). In this cohort of early-stage NSCLC patients, male gender and age >70 years were associated with a worse outcome, but elevated levels of CEA and CYFRA 21-1, and TMI risk were not.


Enhanced solubility and oral bioavailability of γ-tocotrienol using a self-emulsifying drug delivery system (SEDDS).
The aim of this study was to evaluate the in vitro and in vivo performance of γ-tocotrienol (γ-T3) incorporated in a self-emulsifying drug delivery system (SEDDS) and to compare its enhanced performance to a commercially available product, namely Tocovid Suprabio™ (hereafter Tocovid), containing tocotrienols. The solubilization of γ-T3 was tested in a dynamic in vitro lipolysis model followed by in vitro cellular uptake study for the lipolysis products. In addition, in vitro uptake studies using Caco2 cells were conducted at different concentrations of γ-T3 prepared as SEDDS, Tocovid, or mixed micelles. γ-T3 incorporated in SEDDS or Tocovid was orally administered to rats at different doses and absolute oral bioavailability from both formulations were determined. The dynamic in vitro lipolysis experiment showed about two fold increase in the solubilization of γ-T3 prepared as SEDDS compared to Tocovid, which correlated with higher cellular uptake in the subsequent uptake studies. In vitro cellular uptake and in vivo oral bioavailability studies have shown a twofold increase in the cellular uptake and oral bioavailability of γ-T3 incorporated in SEDDS compared to Tocovid as a result of improvement in its solubility and passive uptake as confirmed by in vitro studies. In conclusion, incorporation of γ-T3 in SEDDS formulation enhanced γ-T3 solubilization and passive permeability, thus its cellular uptake and oral bioavailability when compared to Tocovid.


Gene function analysis by artificial microRNAs in Physcomitrella patens.
MicroRNAs (miRNAs) are ~21 nt long small RNAs transcribed from endogenous MIR genes which form precursor RNAs with a characteristic hairpin structure. miRNAs control the expression of cognate target genes by binding to reverse complementary sequences resulting in cleavage or translational inhibition of the target RNA. Artificial miRNAs (amiRNAs) can be generated by exchanging the miRNA/miRNA sequence of endogenous MIR precursor genes, while maintaining the general pattern of matches and mismatches in the foldback. Thus, for functional gene analysis amiRNAs can be designed to target any gene of interest. During the last decade the moss Physcomitrella patens emerged as a model plant for functional gene analysis based on its unique ability to integrate DNA into the nuclear genome by homologous recombination which allows for the generation of targeted gene knockout mutants. In addition to this, we developed a protocol to express amiRNAs in P. patens that has particular advantages over the generation of knockout mutants and might be used to speed up reverse genetics approaches in this model species.


The potential role of serotonin reuptake inhibitors in the treatment of obsessive compulsive disorder.
The authors examine the role of the serotonergic system in the modulation or genesis of obsessive compulsive disorder and the potential for the use of the new serotonin (5-HT) reuptake inhibitors in its treatment. The data available are still preliminary, but are accumulating rapidly, and appear to hold promise in helping to understand this disorder and treat it more effectively. An overview of the data available is presented to assist in the planning of future research in this subject area. At least six major questions need to be addressed by future work: (1) What are the major hypotheses regarding the involvement of the serotonin system in the modulation or genesis of OCD? (2) Is there a serotonin system defect associated with OCD? (3) Is a serotonin system defect the underlying cause of OCD? (4) Are serotonin reuptake inhibitors effective in the treatment of OCD? (5) Are the effects of the serotonin reuptake inhibitors that are effective in treating OCD mediated via the serotonin system or via another mechanism? and (6) Are these drugs worth pursuing as treatment of OCD?


BigSeqKit: a parallel Big Data toolkit to process FASTA and FASTQ files at scale.
High-throughput sequencing technologies have led to an unprecedented explosion in the amounts of sequencing data available, which are typically stored using FASTA and FASTQ files. We can find in the literature several tools to process and manipulate those type of files with the aim of transforming sequence data into biological knowledge. However, none of them are well fitted for processing efficiently very large files, likely in the order of terabytes in the following years, since they are based on sequential processing. Only some routines of the well-known seqkit tool are partly parallelized. In any case, its scalability is limited to use few threads on a single computing node. Our approach, BigSeqKit, takes advantage of a high-performance computing-Big Data framework to parallelize and optimize the commands included in seqkit with the aim of speeding up the manipulation of FASTA/FASTQ files. In this way, in most cases, it is from tens to hundreds of times faster than several state-of-the-art tools. At the same time, our toolkit is easy to use and install on any kind of hardware platform (local server or cluster), and its routines can be used as a bioinformatics library or from the command line. BigSeqKit is a very complete and ultra-fast toolkit to process and manipulate large FASTA and FASTQ files. It is publicly available at https://github.com/citiususc/BigSeqKit.


Predictors for drug effects with brain disease: Shed new light from EEG parameters to brain connectomics.
Though researchers spent a lot of effort to develop treatments for neuropsychiatric disorders, the poor translation of drug efficacy data from animals to human hampered the success of these therapeutic approaches in human. Pharmaceutical industry is challenged by low clinical success rates for new drug registration. To maximize the success in drug development, biomarkers are required to act as surrogate end points and predictors of drug effects. The pathology of brain disease could be in part due to synaptic dysfunction. Electroencephalogram (EEG), generating from the result of the postsynaptic potential discharge between cells, could be a potential measure to bridge the gaps between animal and human data. Here we discuss recent progress on using relevant EEG characteristics and brain connectomics as biomarkers to monitor drug effects and measure cognitive changes on animal models and human in real-time. It is expected that the novel approach, i.e. EEG connectomics, will offer a deeper understanding on the drug efficacy at a microcirculatory level, which will be useful to support the development of new treatments for neuropsychiatric disorders.


Safety of potassium-bearing citrate in patients with renal transplantation: A case report.
Urinary lithiasis is one of severe postoperative complications in patients undergoing renal transplantation, possibly leading to anuria, urinary infection, or even acute renal failure. Potassium sodium hydrogen citrate (PSHC), a potassium-bearing citrate, is commonly prescribed to prevent stone formation. A 25-year-old man (patient 1) and a 31-year-old man (patient 2) receiving renal transplantation for end-stage renal disease (ESRD) were enrolled in this study. They were given 10 g/day of PSHC granules from the ninth day to the 17th day after surgery. Patient 1 presented chest tightness, nausea, muscle weakness, and ascending paralysis on the 10th day. Patient 2 presented weak waves on EGG on the 17th day. Moreover, their serum potassium concentrations (SPCs) were 7.67 and 6.05 mmol/L, respectively. Acute hyperkalemia. Hemo-filtration was performed for patient 1, while patient 2 received 10% calcium gluconate 10 mL, 5% NaHCO3 125 mL, and 10% glucose 500 mL with the addition of 10 units of insulin through intravenous drip. Their SPCs dropped to the normal range. Physicians should pay close attentions to potential risks caused by PSHC, and monitor the SPCs to minimize the occurrence of hyperkalemia.


HIV-1 Matrix Trimerization-Impaired Mutants Are Rescued by Matrix Substitutions That Enhance Envelope Glycoprotein Incorporation.
The matrix (MA) domain of HIV-1 Gag plays key roles in virus assembly by targeting the Gag precursor to the plasma membrane and directing the incorporation of the viral envelope (Env) glycoprotein into virions. The latter function appears to be in part dependent on trimerization of the MA domain of Gag during assembly, as disruption of the MA trimer interface impairs Env incorporation. Conversely, many MA mutations that impair Env incorporation can be rescued by compensatory mutations in the trimer interface. In this study, we sought to investigate further the biological significance of MA trimerization by isolating and characterizing compensatory mutations that rescue MA trimer interface mutants with severely impaired Env incorporation. By serially propagating MA trimerization-defective mutants in T cell lines, we identified a number of changes in MA, both within and distant from the trimer interface. The compensatory mutations located within or near the trimer interface restored Env incorporation and particle infectivity and permitted replication in culture. The structure of the MA lattice was interrogated by measuring the cleavage of the murine leukemia virus (MLV) transmembrane Env protein by the viral protease in MLV Env-pseudotyped HIV-1 particles bearing the MA mutations and by performing crystallographic studies of in vitro-assembled MA lattices. These results demonstrate that rescue is associated with structural alterations in MA organization and rescue of MA domain trimer formation. Our data highlight the significance of the trimer interface of the MA domain of Gag as a critical site of protein-protein interaction during HIV-1 assembly and establish the functional importance of trimeric MA for Env incorporation.IMPORTANCE The immature Gag lattice is a critical structural feature of assembling HIV-1 particles, which is primarily important for virion formation and release. While Gag forms a hexameric lattice, driven primarily by the capsid domain, the MA domain additionally trimerizes where three Gag hexamers meet. MA mutants that are defective for trimerization are deficient for Env incorporation and replication, suggesting a requirement for trimerization of the MA domain of Gag in Env incorporation. This study used a gain-of-function, forced viral evolution approach to rescue HIV-1 mutants that are defective for MA trimerization. Compensatory mutations that rescue virus replication do so by restoring Env incorporation and MA trimer formation. This study supports the importance of MA domain trimerization in HIV-1 replication and the potential of the trimer interface as a therapeutic target.


[Outbreak of cryptosporidiosis in Guadarrama (Autonomous Community of Madrid)].
In spring of 1998, an acute gastroenteritis outbreak, which mainly affected preschool children, took place in Guadarrama (Community of Madrid, Spain). A descriptive and analytical (cases and controls) study of the outbreak was done. Stool samples of the cases were taken to microbiological investigation. The tap water getting system of the municipality was inspected and samples were collected to microbiological analysis. Twenty one children from 0 to 5 years-old had liquid diarrhea which lasted more than five days. A significant statistical association was found within tap water consumption and gastroenteritis [OR = 5.73 (1.18-43.30); p < 0.05]. It did not have association with other variables investigated. Cryptosporidium parvum (Cp) oocysts were detected in stool samples of eight cases. Deficiencies were observed in the tap water getting and treatment system of the municipality but no oocysts were found in the analysis of the water. The results of the epidemiological investigation support that the source of the gastroenteritis outbreak was tap water, likely contaminated with Cp. While this parasite was not detected in the analysis of the water, it is explainable for the difficulties which this technique involves. In our review similar descriptions of outbreaks have not been found in Spain, but it seems that this reports are not infrequent in neighbour countries. Therefore, Cp should be borne in mind in diagnostic and management of these situations.


N-acetyltransferase 2 phenotype, occupation, and bladder cancer risk: results from the EPIC cohort.
An association between N-acetyltransferase 2 (NAT2) slow acetylation and bladder cancer has been consistently observed in epidemiologic studies. However, evidence has been mainly derived from case-control studies and was sparse from cohort studies. We evaluated the association between NAT2 slow acetylation and bladder cancer in a case-control study nested in the European Prospective Investigation into Cancer and Nutrition. Exposure to aromatic amines and polycyclic aromatic hydrocarbons (PAH) could be assessed for 754 cases and 833 controls for whom occupational information was documented. A semiquantitative job-exposure matrix was applied to at-risk occupations to estimate the exposure as low, medium, or high based on tertiles of the distribution of the exposure score in controls. Using a comprehensive genotyping, NAT2 acetylation status could be categorized from 6-single-nucleotide polymorphism genotypes as slow or fast in 607 cases and 695 controls with DNA from archived blood samples. Occupational exposure to aromatic amines and PAH was associated with an increased bladder cancer risk [upper tertile of the distribution of the exposure score: OR = 1.37; 95% confidence interval (CI), 1.02-1.84, and OR = 1.50; 95% CI, 1.09-2.05, respectively]. NAT2 slow acetylation did not modify these risk estimates and was not itself associated with bladder cancer risk (OR = 1.02; 95% CI, 0.81-1.29). These findings confirm established or suspected occupational risk factors but not the anticipated role of NAT2 slow acetylation in bladder cancer. No interaction was detected between NAT2 and any exposure of interest, including smoking. Genetic testing for NAT2 would be inappropriate in occupational settings.


Breeding success of a marine central place forager in the context of climate change: A modeling approach.
In response to climate warming, a southward shift in productive frontal systems serving as the main foraging sites for many top predator species is likely to occur in Subantarctic areas. Central place foragers, such as seabirds and pinnipeds, are thus likely to cope with an increase in the distance between foraging locations and their land-based breeding colonies. Understanding how central place foragers should modify their foraging behavior in response to changes in prey accessibility appears crucial. A spatially explicit individual-based simulation model (Marine Central Place Forager Simulator (MarCPFS)), including bio-energetic components, was built to evaluate effects of possible changes in prey resources accessibility on individual performances and breeding success. The study was calibrated on a particular example: the Antarctic fur seal (Arctocephalus gazella), which alternates between oceanic areas in which females feed and the land-based colony in which they suckle their young over a 120 days rearing period. Our model shows the importance of the distance covered to feed and prey aggregation which appeared to be key factors to which animals are highly sensitive. Memorization and learning abilities also appear to be essential breeding success traits. Females were found to be most successful for intermediate levels of prey aggregation and short distance to the resource, resulting in optimal female body length. Increased distance to resources due to climate warming should hinder pups' growth and survival while female body length should increase.


Using ADAPT-ITT to Modify a Telephone-Based HIV Prevention Intervention for SMS Delivery: Formative Study.
African American adolescent females are disproportionately affected by sexually transmitted infections (STIs) and HIV. Given the elevated risk of STIs and HIV in African American women, there is an urgent need to identify innovative strategies to enhance the adoption and maintenance of STI and HIV preventive behaviors. Texting is a promising technology for creating preventive maintenance interventions (PMIs) that extend the efficacy of the original intervention. However, little guidance in public health literature is available for developing this type of application. This paper describes a formative pilot study that incorporates user experience methods to design and test PMI texts for Afiya, an original evidence-based intervention (EBI) specifically designed for African American adolescent females. This study aims to describe the adaptation process of health educator-led phone calling to text-based communication. The formative process followed the assessment, decision, adaptation, production, topical experts-integration, training, testing (ADAPT-ITT) framework for adapting EBIs and using them in a new setting, for a new target population or a modified intervention strategy. This study presents the details of how the phases of the ADAPT-ITT framework were applied to the design of the adaptation. An advisory board was constituted from the target population, consisting of 6 African American women aged 18-24 years, participating in formative activities for 12 weeks, and involving components of the PMI design. As Afiya included a telephone-based PMI, developers of the original Afiya phone scripts crafted the initial design of the SMS-based texts and texting protocol. The advisory board participated in the 1-day Afiya workshop, followed by 4 weeks of texting PMI messages and a midcourse focus group, followed by 4 more weeks of texting PMI messages, ultimately ending with a final focus group. At the advisory board's request, this phase included an optional, additional week of text-based PMI messages. The methods provided a rich source of data and insights into the fundamental issues involved when constructing SMS-based PMI for this target population and for this EBI. Prior contact and context are essential as the health educator was identified as a key persona in the process and the messages were situated in the original (workshop) context. Narrative adaptations for personas emerged from advisory board discussions. Suggestions on how to expand the PMI to current, specific social contexts indicated that the use of narrative analysis is warranted. The use of existing EBIs incorporating telephone-based PMI scripts facilitated the initial design of the texts, with a subsequent narrative analysis of the advisory board data providing additional adjustments given the actual context. Additional examination of the advisory board feedback revealed that personas would offer insight into and opportunities for a persona-specific modification of texting narratives.


The anandamide transport inhibitor AM404 reduces ethanol self-administration.
The endocannabinoid system mediates in the pharmacological actions of ethanol and genetic studies link endocannabinoid signaling to alcoholism. Drugs activating cannabinoid CB1 receptors have been found to promote alcohol consumption but their effects on self-administration of alcohol are less clear because of the interference with motor performance. To avoid this problem, a novel pharmacological approach to the study of the contribution of the cannabinoid system in alcoholism may be to use drugs that locally amplify the effects of alcohol on endogenous cannabinoids. In the present study we addressed this model by studying the effects of the anandamide transport inhibitor N-(4-hydroxyphenyl) arachidonoyl-ethanolamide (AM404) on both ethanol self-administration and reinstatement of alcohol-seeking behavior in rats. The results show that AM404 significantly reduced ethanol self-administration in a dose-dependent manner but failed to modify reinstatement for lever pressing induced by the stimulus associated with alcohol. This effect was not due to a motor depressant effect and was not related to a decrease in general motivational state, as it was not effective in other reward paradigms such as lever pressing for a saccharin solution. The mechanism of action of AM404 does not involve cannabinoid CB1 receptors as the CB1-selective antagonist SR141716A did not block the reduction of ethanol self-administration induced by the anandamide uptake blocker. Moreover, 3-(1,1-dimethylheptyl)-(-)-11-hydroxy-delta 8-tetrahydrocannabinol (HU-210), a classical cannabinoid receptor agonist, did not affect ethanol self-administration. The effects of AM404 are not mediated by either vanilloid VR1 receptors or cannabinoid CB2 receptors because it is not antagonized by either the VR1 receptor antagonist capsazepine or the CB2 antagonist AM630. These results indicate that AM404 may be considered as an innovative approach to reduce alcohol consumption.


Fungal treatment for liquid waste containing U(VI) and Th(IV).
Four fungal and one bacterial isolates were isolated from a liquid waste sample of Nuclear Material Authority. Those dried biomasses were screened for uranium (U) and thorium (Th) adsorption efficiency where the most potent isolate was identified according to sequence similarities and phylogenetic analysis as Aspergillus niger LBM 134. Using U or Th synthetic solutions many factors were investigated for controlling the biosorption process to conduct the optimum process conditions (the solution pH, contact time, elemental initial concentration, biomass dosage, and sorption temperature). A. niger LBM 134 dried biomass was examined ESEM-EDX and the FTIR techniques before and after the sorption process, also the data were handled by different kinetics and isothermal models. Application on the real liquid waste revealed that the bio-uptake capacities were 18.5 and 11.1 mg/g for U and Th respectively.


The computer user syndrome.
Most conventional computer hardware systems have been inadequately designed for the functional comfort and efficient usage by human operators. The effects of excyclotorsion, inadequate depression-of-gaze, excessive hand-wrist pronation, and ulnar abduction are discussed. The neurology of ocular movements, synkinesis, saccades, pursuits, and orthopedic hand-wrist biomechanics are reviewed. The interrelationships of these ocular and orthopedic phenomena have been synthesized into a comprehensive hypothesis, in an effort to create a computer configuration which permits a greater integration of the keyboard (tool-usage) with the screen-visualization (product-of-tool-usage), and improves visual feedback.


Recombinant sclerostin antagonizes effects of ex vivo mechanical loading in trabecular bone and increases osteocyte lacunar size.
Sclerostin has emerged as an important regulator of bone mass. We have shown that sclerostin can act by targeting late osteoblasts/osteocytes to inhibit bone mineralization and to upregulate osteocyte expression of catabolic factors, resulting in osteocytic osteolysis. Here we sought to examine the effect of exogenous sclerostin on osteocytes in trabecular bone mechanically loaded ex vivo. Bovine trabecular bone cores, with bone marrow removed, were inserted into individual chambers and subjected to daily episodes of dynamic loading. Cores were perfused with either osteogenic media alone or media containing human recombinant sclerostin (rhSCL) (50 ng/ml). Loaded control bone increased in apparent stiffness over time compared with unloaded bone, and this was abrogated in the presence of rhSCL. Loaded bone showed an increase in calcein uptake as a surrogate of mineral accretion, compared with unloaded bone, in which this was substantially inhibited by rhSCL treatment. Sclerostin treatment induced a significant increase in the ionized calcium concentration in the perfusate and the release of β-CTX at several time points, an increased mean osteocyte lacunar size, indicative of osteocytic osteolysis, and the expression of catabolism-related genes. Human primary osteocyte-like cultures treated with rhSCL also released β-CTX from their matrix. These results suggest that osteocytes contribute directly to bone mineral accretion, and to the mechanical properties of bone. Moreover, it appears that sclerostin, acting on osteocytes, can negate this effect by modulating the dimensions of the lacunocanalicular porosity and the composition of the periosteocyte matrix.


Autism before diagnosis: crying, feeding and sleeping problems in the first two years of life.
To chart early registered regulatory problems (RP) in a representative group of young children with and without autism spectrum disorder (ASD). The target group comprised 208 preschool children with ASD, whose records from the Child Health Centres (CHC) were reviewed regarding numbers of consultations for excessive crying, feeding and sleeping problems. The records from an age- and gender-matched comparison group were obtained from the same CHCs as those of the index children Significant differences between the ASD and comparison groups were found for each domain studied and when domains were collapsed. Two or more consultations had occurred in 44% of the children in the ASD group and in 16% of the comparison group (p < 0.001). No correlations were found with regard to gender, later severity of autism, cognitive level or degree of hyperactivity. Regulatory problems (RP) were much more common in children who later received a diagnosis of ASD. Children with many RP in infancy require attention from CHC and paediatric services and need to be followed with regard to development and family support.


Purification and characterization of a novel class III peroxidase isoenzyme from tea leaves.
A novel, basic (isoelectric point > 10), heme peroxidase isoenzyme (TP; relative molecular weight = 34,660 +/- 10, mean +/- SE) that can account for a significant part of the ascorbate peroxidase activity in tea (Camellia sinensis) leaves has been purified to homogeneity. The ultraviolet/visible absorption spectrum is typical of heme-containing plant peroxidases, with a Soret peak at 406 nm (epsilon = 115 mM-1 cm-1) and an A406/A280 value of 3.4. The enzyme has a high specific activity for ascorbate oxidation (151 mumol min-1 mg-1), with a pH optimum in the range of 4.5 to 5.0. Substrate-specificity studies have revealed significant differences between TP and other class III peroxidases, as well as similarities with class I ascorbate peroxidases. TP, like ascorbate peroxidase, exhibits a preference for ascorbate over guaiacol, whereas other class III isoenzymes are characterized by 2-orders-of-magnitude higher activity for guaiacol than for ascorbate. TP also forms an unstable porphyrin pi cation radical-type compound I, which is converted to compound II within approximately 2 min in the absence of added reductant. Amino acid sequence data show TP to be the first example, to our knowledge, of a class III peroxidase with a high specificity for ascorbate as an electron donor.


Coronary Heart Disease (CHD) Risk Factors and Metabolic Syndrome in HIV-Positive Drug Users in Miami.
The frequency of coronary heart disease (CHD) is increasing among HIV seropositive persons. This phenomenon may be related to HIV disease itself, the use of antiretroviral medications and increased length of survival, or the synergism of these factors. In this study we have calculated the 10-year CHD risk estimate and the prevalence of metabolic syndrome in a cohort of 118 HIV seropositive chronic drug users, including those who are on HAART with or without protease inhibitors (PI). The results showed that the 10-year coronary heart disease risk among the HIV seropositive drug users was 4.8 ± 5.7, which is within the range of results published for other HIV infected cohorts. The 10-year CHD risk was significantly higher in men (5.9±6.1, p<0.001) than in women (1.7±2.4), due to their gender and the pre-menopausal mean age of the women (39.4±7.3 years of age), despite a significantly higher rate of abdominal obesity (54.8% in women vs. 8.1% in men, p<0.001) and lower HDL (61.3% in women vs. 40% in men, p=0.042). The rate of metabolic syndrome among our female HIV seropositive drug users was significantly higher (29% vs 10.3%, p=0.013) compared to men (10.3%). Participants with metabolic syndrome had a significantly higher 10-year CHD risk (27.8% vs. 10.2%, p=0.041) and higher mean BMI (28.6 ± 4.1 vs. 24.2±4, p<0.001) than those without the syndrome. The predominant proportion of the cohort had a high viral load, suggesting that their use of illicit drugs has an influence on either adherence or effectiveness of antiretroviral medication. Increased viral load was significantly associated with metabolic syndrome (OR=2.23, 95% CI:1.12, 4.47; p=0.023), high fasting glucose (OR=1.61, 95% CI: 1.02, 2.55; p=0.042) and low HDL levels (OR=1.41, 95% CI: 1.01, 1.98; p=0.046), after controlling for age gender, smoking, PI exposure, BMI and CD4. HAART with or without PI did not significantly impact the 10-year CHD risk estimate or metabolic syndrome in this cohort. The estimated effect of PI, however, was positively and significantly related to triglyceride levels (effect estimate=95.81; 95% CI:39.40, 152.21; p<0.01) after controlling for age, gender, smoking, viral load, CD4 cell count and BMI. Heavy use of cigarettes and crack/cocaine was inversely associated with obesity (OR=0.84, 95% CI:0.67, 0.99; p=0.049; OR=0.43, 95% CI:0.19, 0.98; p=0.044, respectively), while use of marijuana tended to be associated with increased central obesity (p=0.08). Heavy cigarette smoking was significantly associated with low HDL (OR=3.06, 95% CI:1.18; 7.95, p=0.02). The significant association of higher viral load with CHD risk indicates that controlling viral load may be important in reducing CHD risk in HIV infected drug users.


A study of the psychosocial characteristics of patients in a geriatric rehabilitation unit in Israel.
This study characterizes the demographic backgrounds of patients in an Israeli geriatric rehabilitation unit, determines the factors associated with their family relationships and the instrumental support they received, and emphasizes the importance of social roles as a personal resource. The study population consisted of 336 low-income Jews, all of whom were immigrants. Virtually all of the subjects had a small, close support network composed mainly of their children and spouses. Their children were the most important source of instrumental support during their hospitalization. The subjects' sources of instrumental support prior to hospitalization varied, depending upon their age, gender, marital status, and social roles. Social exchange theory provided a framework for explaining their social roles. Factors found to be predictors of good family relationships were marital status, living arrangements, instrumental support, social roles, and educational level.


Simple and efficient defect-tailored fiber-based UV-VIS broadband white light generation.
We propose and demonstrate a facile approach for ultraviolet-visible broadband generation from a sapphire crystal core-borosilicate glass cladding hybrid fiber using a laser-heated pedestal growth technique. Considerable formation of F- and F(2)-type color emitters is effectively facilitated by Ti(4+) ions and Al(3+) vacancies, retaining efficient luminescence and high crystallinity of the sapphire core. These color centers intensify the ultraviolet, blue, and green emissions at 370, 450, and 540 nm, whereas the 650-nm red emission is contributed by Cr(3+) in the octahedral sites of the corundum structure. Over 1-mW white light with an optical-to-optical efficiency of up to nearly 5% and 1931 Commission International de l'Eclairage chromaticity coordinate of (0.287, 0.333) is achieved under 325-nm excitation.


On the effect of ionizing radiation upon the retina in man and animals.
The phenomenon of light flashes observed by the American cosmonauts on exposure to primary cosmic radiation during translunar flights, and also perceived by other observers on exposure of the eye to neutrons from accelerators, is indeed one of the latest manifestations of the sensitivity of the retina to very high energy radiation. The usual interpretations of this phenomenon are based either on the hypothesis of Vavilov-Cherenkov radiation in the ocular media, or on the direct stimulation of photoreceptors in the retina itself. The data available are submitted to adequate analysis, the experimental materials concerning the radiobiology of the retina are generalized, and a conclusion is drawn as to the most probable mechanism of light flashes considered as signals damaging the outer segments of the rods by particles arising from nuclear reactions in the retina.


The role of heritable and dietary factors in the sexual signal of a Hispaniolan Anolis lizard, Anolis distichus.
The diversity of sexual signals is astounding, and divergence in these traits is believed to be associated with the early stages of speciation. An increasing number of studies also suggest a role for natural selection in driving signal divergence for effective transmission in heterogeneous environments. Both speciation and adaptive divergence, however, are contingent on the sexual signal being heritable, yet this often remains assumed and untested. It is particularly critical that the heritability of carotenoid-based sexual signals is investigated because such traits may instead be phenotypically plastic indicators of an individual's quality that exhibit no or little heritable variation. We present the first study to investigate the relative contribution of genetic and environmental factors to the striking diversity of dewlap color and pattern in Anolis lizards. Using a breeding experiment with Anolis distichus populations exhibiting different dewlap phenotypes, we raise F1 offspring in a common garden experiment to assess whether dewlap color is inherited. We follow this with carotenoid supplementation to investigate the influence of dietary pigments to dewlap color variation. We find significant differences in several aspects of dewlap color and pattern to persist to the F1 generation (fathers: N = 19; F1 males: N = 50; P < 0.01) with no change in dewlap phenotype with carotenoid supplementation (N = 52; P > 0.05). These results strongly support that genetic differences underlie dewlap color variation, thereby satisfying a key requirement of natural selection. Our findings provide an important stepping-stone to understanding the evolution of an incredibly diverse signal important for sexual selection and species recognition.


Coactivator and corepressor gene expression in rat cerebellum during postnatal development and the effect of altered thyroid status.
Thyroid hormone (TH) plays an important role in the postnatal development of the rodent cerebellum, particularly within the first 2 weeks of postnatal life. This action is exerted through the regulation of specific genes during development and is mediated by coactivator and corepressor proteins that determine transcriptional repression or activation, respectively. Thus, we hypothesized that the effect of TH on rodent cerebellar development could be influenced by the relative amounts of coactivator and corepressor proteins in vivo. These ratios might be modulated in an age-specific manner and/or by hormones to generate the "critical period" of TH action. To examine this hypothesis, we cloned rat complementary DNA fragments corresponding to coactivators (SRC1, TIF2 and TRAM1) and corepressors (N-CoR and SMRT), and studied the ontogenic changes in their corresponding messenger RNAs in rat cerebellum of normal and hypothyroid rats during postnatal development, using a RNase protection assay. We found an increased expression of SRC1 and TIF2, as well as of N-CoR, during rat cerebellar development but no change in the expression of SMRT and TRAM1 genes. However, thyroid hormone status did not affect the expression of coactivator and corepressor genes in the cerebellum. These results indicate that coactivator and corepressor messenger RNAs exhibit differential expression through cerebellar development but are not regulated by TH during this period.


Vascular cell adhesion molecule-1 expression in human intestinal microvascular endothelial cells is regulated by PI 3-kinase/Akt/MAPK/NF-kappaB: inhibitory role of curcumin.
Endothelial activation and surface expression of cell adhesion molecules (CAMs) is critical for binding and recruitment of circulating leukocytes in tissues during the inflammatory response. Endothelial CAM expression plays a critical role in the intestinal microvasculature in inflammatory bowel disease (IBD), as blockade of leukocyte alpha4-integrin binding by gut endothelial CAM ligands has therapeutic benefit in IBD. Mechanisms underlying expression of vascular cell adhesion molecule (VCAM)-1, a ligand for alpha4-integrin in primary cultures of human intestinal microvascular endothelial cells (HIMEC) has not been defined. We investigated the effect of curcumin, phosphatidylinositol 3-kinase (PI 3-kinase)/protein kinase B (Akt), and mitogen-activated protein kinase (MAPK) inhibitors on VCAM-1 expression and function in HIMEC. CAM expression was assessed and HIMEC-leukocyte adhesion was visualized under static and flow conditions. Western blotting and in vitro kinase assays were used to assess Akt and MAPK activation. Nuclear factor-kappaB (NF-kappaB) activation and nuclear translocation of its p65 subunit were determined. Tumor necrosis factor (TNF)-alpha/lipopolysaccharide (LPS)-induced VCAM-1 expression in HIMEC was suppressed by Akt small-interfering RNA, curcumin, and inhibitors of NF-kappaB (SN-50), p38 MAPK (SB-203580) and PI 3-kinase/Akt (LY-294002). VCAM-1 induction was partially suppressed by p44/42 MAPK (PD-098059) but unaffected by c-Jun NH2-terminal kinase (SP-600125) inhibition. Curcumin inhibited Akt/MAPK/NF-kappaB activity and prevented nuclear translocation of the p65 NF-kappaB subunit following TNF-alpha/LPS. At physiological shear stress, curcumin attenuated leukocyte adhesion to TNF-alpha/LPS-activated HIMEC monolayers. In conclusion, curcumin inhibited the expression of VCAM-1 in HIMECs through blockade of Akt, p38 MAPK, and NF-kappaB. Curcumin may represent a novel therapeutic agent targeting endothelial activation in IBD.


CGI-58 knockdown in mice causes hepatic steatosis but prevents diet-induced obesity and glucose intolerance.
Mutations of Comparative Gene Identification-58 (CGI-58) in humans cause triglyceride (TG) accumulation in multiple tissues. Mice genetically lacking CGI-58 die shortly after birth due to a skin barrier defect. To study the role of CGI-58 in integrated lipid and energy metabolism, we utilized antisense oligonucleotides (ASOs) to inhibit CGI-58 expression in adult mice. Treatment with two distinct CGI-58-targeting ASOs resulted in ∼80-95% knockdown of CGI-58 protein expression in both liver and white adipose tissue. In chow-fed mice, ASO-mediated depletion of CGI-58 did not alter weight gain, plasma TG, or plasma glucose, yet raised hepatic TG levels ∼4-fold. When challenged with a high-fat diet (HFD), CGI-58 ASO-treated mice were protected against diet-induced obesity, but their hepatic contents of TG, diacylglycerols, and ceramides were all elevated, and intriguingly, their hepatic phosphatidylglycerol content was increased by 10-fold. These hepatic lipid alterations were associated with significant decreases in hepatic TG hydrolase activity, hepatic lipoprotein-TG secretion, and plasma concentrations of ketones, nonesterified fatty acids, and insulin. Additionally, HFD-fed CGI-58 ASO-treated mice were more glucose tolerant and insulin sensitive. Collectively, this work demonstrates that CGI-58 plays a critical role in limiting hepatic steatosis and maintaining hepatic glycerophospholipid homeostasis and has unmasked an unexpected role for CGI-58 in promoting HFD-induced obesity and insulin resistance.


Vaccine Vectors Harnessing the Power of Cytomegaloviruses.
Cytomegalovirus (CMV) species have been gaining attention as experimental vaccine vectors inducing cellular immune responses of unparalleled strength and protection. This review outline the strengths and the restrictions of CMV-based vectors, in light of the known aspects of CMV infection, pathogenicity and immunity. We discuss aspects to be considered when optimizing CMV based vaccines, including the innate immune response, the adaptive humoral immunity and the T-cell responses. We also discuss the antigenic epitopes presented by unconventional major histocompatibility complex (MHC) molecules in some CMV delivery systems and considerations about routes for delivery for the induction of systemic or mucosal immune responses. With the first clinical trials initiating, CMV-based vaccine vectors are entering a mature phase of development. This impetus needs to be maintained by scientific advances that feed the progress of this technological platform.


Structural, functional, and mutational analysis of the NblA protein provides insight into possible modes of interaction with the phycobilisome.
The enormous macromolecular phycobilisome antenna complex (>4 MDa) in cyanobacteria and red algae undergoes controlled degradation during certain forms of nutrient starvation. The NblA protein (approximately 6 kDa) has been identified as an essential component in this process. We have used structural, biochemical, and genetic methods to obtain molecular details on the mode of action of the NblA protein. We have determined the three-dimensional structure of the NblA protein from both the thermophilic cyanobacterium Thermosynechococcus vulcanus and the mesophilic cyanobacterium Synechococcus elongatus sp. PCC 7942. The NblA monomer has a helix-loop-helix motif which dimerizes into an open, four-helical bundle, identical to the previously determined NblA structure from Anabaena. Previous studies indicated that mutations to NblA residues near the C terminus impaired its binding to phycobilisome proteins in vitro, whereas the only mutation known to affect NblA function in vivo is located near the protein N terminus. We performed random mutagenesis of the S. elongatus nblA gene which enabled the identification of four additional amino acids crucial for NblA function in vivo. This data shows that essential amino acids are not confined to the protein termini. We also show that expression of the Anabaena nblA gene complements phycobilisome degradation in an S. elongatus NblA-null mutant despite the low homology between NblAs of these cyanobacteria. We propose that the NblA interacts with the phycobilisome via "structural mimicry" due to similarity in structural motifs found in all phycobiliproteins. This suggestion leads to a new model for the mode of NblA action which involves the entire NblA protein.


[The healing effects of vitamin E with corticosteroid and vitamin E alone on nerve healing in rats with traumatic facial palsy].
We aimed to document the effects of vitamin E and vitamin E plus corticosteroid on nerve healing in rats. Twenty-four Wistar Albino type female rats were used in the study. The rats were divided in to three groups, each with eight rats. Group 1 was given methylprednisolone and Vitamin E, group 2 was given Vitamin E alone and group 3 was the control group. The buccal branch of the facial nerve was found under anesthesia in all rats. After nerve identification, the stimulation thresholds were assessed by Xomed Trace Nerve Integrity Monitor NIM-2. The buccal branch of each facial nerve in group 1 and 2 was traumatized by a vascular clamp for an hour. After trauma the stimulus thresholds were identified again. The nerve was not traumatized in group 3. Group 1 was given 0.1 mg/kg/day vitamin E intramuscular (i.m) and 1 mg/kg/day methylprednisolone i.m for one month, group 2 was given 0.1 mg/kg/day vitamin E i.m for one month and the control group was given 2 cc 0.09% NaCl subcutaneously every other day for one month. At the end of a month the stimulus thresholds were measured again and compared statistically. Mean stimulus threshold was 0.12 mA for all of the groups at the beginning of the experiment. After trauma mean stimulus threshold was 0.48 mA for the group 1 and 2. At the end of one month stimulus threshold was 0.22 mA for the group 1 and 0.33 mA for the group 2. The stimulus threshold in the control group was measured as 0.12 mA. We concluded that the vitamin E has a positive effect on nerve healing and corticosteroid enhances its effect.


A CLDN18.2-Targeting Bispecific T Cell Co-Stimulatory Activator for Cancer Immunotherapy.
Co-stimulatory receptor agonist antibodies have shown promising antitumor efficacy in preclinical models. However, their clinical development lags due to systemic or local adverse effects of non-specific T cell activation. Utilization of a bispecific antibody format to reduce off-tumor immune activation is a focus of co-stimulatory receptor agonist antibody design. In this study, a bispecific antibody with anti-CLDN18.2 and anti-CD28 moieties was produced. Its T cell costimulation ability was evaluated in T cell coculture assay in vitro. Its safety and anti-tumor efficacy were explored in mouse tumor models. Anti-CLDN18.2-anti-CD28 bispecific antibody could co-stimulate T cells and increase the expression of effector cytokines in a CLDN18.2-dependent manner. Treatment of anti-CLDN18.2-anti-CD28 could reduce tumor burden and increase tumor-infiltrated T cells. Immunosuppressive cells including tumor-associated macrophages and myeloid-derived suppressor cells were also reduced without systemic adverse effects. This work provided proof-of-concept evidence for a new strategy to develop a bispecific co-stimulatory activator for treating CLDN18.2+ tumors.


Rhodamine-naphthalimide demonstrated a distinct aggregation-induced emission mechanism: elimination of dark-states via dimer interactions (EDDI).
Rhodamine B-naphthalimide (RhB-Naph) demonstrated a distinct aggregation-induced emission (AIE) mechanism, different from the restriction of intramolecular rotations or vibrations as in traditional AIE molecules. The monomers of RhB-Naph were non-emissive, due to the presence of a dark S1 state. Upon molecular aggregation, intermolecular interactions significantly altered the electronic properties of RhB-Naph, leading to the formation of a bright S1 state and endowing RhB-Naph with notable AIE properties. Besides, we demonstrated that RhB-Naph enabled the development of a solid-state three-color fluorescent switch upon multi-external stimuli.


The physics of surfactant dissolution.
We have examined the early stages of surfactant dissolution and mesophase formation using a dimer-solvent model with phase behaviour representative of surfactant-water systems. We use an orientational order parameter to characterize systematically the appearance of mesophases. We find the process is diffusion controlled, and the appearance of mesophases is governed adiabatically by the equilibrium phase diagram from the earliest point at which the orientational order parameter can reliably distinguish between mesophases, when only a few repeat spacings of the mesophase microstructures are present. In real systems, such a stage would correspond to times of the order of a few microseconds after the initial contact.


An investigation into the spatial relationship between complexity and motility within the oesophagus.
Nonlinear analysis techniques have recently been used in the characterization of complex physiological signals seen in pathological disorders such as epilepsy and cardiac fibrillation. In this study a series of controlled swallows from an asymptomatic demonstration group was investigated using oesophageal manometry. The nonlinear measure of complexity, largest Lyapunov exponents and phase portraits were then used to explore the complexity of motility patterns at different points within the oesophagus. Results indicate greater complexity within the region of the striated muscle in the upper oesophagus than that observed within the region of smooth muscle in the lower oesophagus. Phase portraits showed that manometry patterns within the asymptomatic demonstration group could be quite different, highlighting the problems in clinical diagnosis. The characterization of motility disorders associated with complex manometry patterns such as diffuse oesophageal spasm (DOS) and nonspecific motility disorder (NOMD) still represents a diagnostic challenge. The use of nonlinear techniques enabling the quantitative and qualitative measurement of oesophageal complexity is considered in the classification of such disorders.


[Efficacy of reconstruction for congenital microphthalmos].
To investigate the therapeutic procedures and their efficiencies for the treatment of congenital microphthalmos during or after the developing period. It was a retrospective case series study. Eighteen cases with congenital microphthalmos, aged less than 12 years, underwent self inflating osmotic hydrogel expanders. Among them, 11 cases were treated by implantation with sphere self-inflating hydrogel expanders into the retrobulbar space and 5 cases had implantations into the conjunctival socket. The remaining 2 cases were treated with implantation of injectable self-inflating hydrogel pellet expanders. Twenty eight cases with congenital microphthalmos, aged more than 12 years, accompanied with other deformities, were treated with proper reconstruction surgery. Fourteen cases were treated with medial canthoplasty, 12 cases were treated with medial and lateral canthoplasty, 3 cases were treated with ptosis correction and medial or lateral canthoplasty and 2 cases with socket reconstruction. T-test was used for the comparison between groups. Sixteen cases obtained satisfactory results without any complications. In one case, the hydrogel expanders migrated inferiorly in the orbit and underwent adjustment. In another case, although the hydrogel expander prolapsed from the socket, the socket was still expanded. In the developing period, the length of palpebral fissure was (15.83 ± 1.87) mm before the operation, and that made much wider after the operation (18.67 ± 1.74) mm. There were statistically significant differences between them (t = -4.029, P = 0.01). The height of palpebral fissure was (1.33 ± 0.67) mm and (6.00 ± 0.58) mm before and after the operation, respectively, which were statistically significant (t = -5.813, P = 0.00). Patients who aged more than 12 years and underwent eyelids and eye socket reconstruction, also obtained satisfactory appearance. Patients with congenital microphthalmos could obtain the development of orbit and satisfactory appearance by reconstruction treatment according to different period of development.


Do adjunctive flap-monitoring technologies impact clinical decision making? An analysis of microsurgeon preferences and behavior by body region.
Multiple perfusion assessment technologies exist to identify compromised microvascular free flaps. The effectiveness, operability, and cost of each technology vary. The authors investigated surgeon preference and clinical behavior with several perfusion assessment technologies. A questionnaire was sent to members of the American Society for Reconstructive Microsurgery concerning perceptions and frequency of use of several technologies in varied clinical situations. Demographic information was also collected. Adjusted odds ratios were calculated using multinomial logistic regression accounting for clustering of similar practices within institutions/regions. The questionnaire was completed by 157 of 389 participants (40.4 percent response rate). Handheld Doppler was the most commonly preferred free flap-monitoring technology (56.1 percent), followed by implantable Doppler (22.9 percent) and cutaneous tissue oximetry (16.6 percent). Surgeons were significantly more likely to opt for immediate take-back to the operating room when presented with a concerning tissue oximetry readout compared with a concerning handheld Doppler signal (OR, 2.82; p < 0.01), whereas other technologies did not significantly alter postoperative management more than simple handheld Doppler. Clinical decision making did not significantly differ by demographics, training, or practice setup. Although most surgeons still prefer to use standard handheld Doppler for free flap assessment, respondents were significantly more likely to opt for immediate return to the operating room for a concerning tissue oximetry reading than an abnormal Doppler signal. This suggests that tissue oximetry may have the greatest impact on clinical decision making in the postoperative period.


Comparative Transcriptomics of Seasonal Phenotypic Flexibility in Two North American Songbirds.
Phenotypic flexibility allows organisms to reversibly alter their phenotypes to match the changing demands of seasonal environments. Because phenotypic flexibility is mediated, at least in part, by changes in gene regulation, comparative transcriptomic studies can provide insights into the mechanistic underpinnings of seasonal phenotypic flexibility, and the extent to which regulatory responses to changing seasons are conserved across species. To begin to address these questions, we sampled individuals of two resident North American songbird species, American goldfinch (Spinus tristis) and black-capped chickadee (Poecile atricapillus) in summer and winter to measure seasonal variation in pectoralis transcriptomic profiles and to identify conserved and species-specific elements of these seasonal profiles. We found that very few genes exhibited divergent responses to changes in season between species, and instead, a core set of over 1200 genes responded to season concordantly in both species. Moreover, several key metabolic pathways, regulatory networks, and gene functional classes were commonly recruited to induce seasonal phenotypic shifts in these species. The seasonal transcriptomic responses mirror winter increases in pectoralis mass and cellular metabolic intensity documented in previous studies of both species, suggesting that these seasonal phenotypic responses are due in part to changes in gene expression. Despite growing evidence of muscle nonshivering thermogenesis (NST) in young precocial birds, we did not find strong evidence of upregulation of genes putatively involved in NST during winter in either species, suggesting that seasonal modification of muscular NST is not a prominent contributor to winter increases in thermogenic capacity for adult passerine birds. Together, these results provide the first comprehensive overview of potential common regulatory mechanisms underlying seasonally flexible phenotypes in wild, free-ranging birds.


The chaperone protein SmgGDS interacts with small GTPases entering the prenylation pathway by recognizing the last amino acid in the CAAX motif.
Ras family small GTPases localize at the plasma membrane, where they can activate oncogenic signaling pathways. Understanding the mechanisms that promote membrane localization of GTPases will aid development of new therapies to inhibit oncogenic signaling. We previously reported that SmgGDS splice variants promote prenylation and trafficking of GTPases containing a C-terminal polybasic region and demonstrated that SmgGDS-607 interacts with nonprenylated GTPases, whereas SmgGDS-558 interacts with prenylated GTPases in cells. The mechanism that SmgGDS-607 and SmgGDS-558 use to differentiate between prenylated and nonprenylated GTPases has not been characterized. Here, we provide evidence that SmgGDS-607 associates with GTPases through recognition of the last amino acid in the CAAX motif. We show that SmgGDS-607 forms more stable complexes in cells with nonprenylated GTPases that will become geranylgeranylated than with nonprenylated GTPases that will become farnesylated. These binding relationships similarly occur with nonprenylated SAAX mutants. Intriguingly, farnesyltransferase inhibitors increase the binding of WT K-Ras to SmgGDS-607, indicating that the pharmacological shunting of K-Ras into the geranylgeranylation pathway promotes K-Ras association with SmgGDS-607. Using recombinant proteins and prenylated peptides corresponding to the C-terminal sequences of K-Ras and Rap1B, we found that both SmgGDS-607 and SmgGDS-558 directly bind the GTPase C-terminal region, but the specificity of the SmgGDS splice variants for prenylated versus nonprenylated GTPases is diminished in vitro. Finally, we present structural homology models and data from functional prediction software to define both similar and unique features of SmgGDS-607 when compared with SmgGDS-558.


Estimating the prevalence of limb loss in the United States: 2005 to 2050.
To estimate the current prevalence of limb loss in the United States and project the future prevalence to the year 2050. Estimates were constructed using age-, sex-, and race-specific incidence rates for amputation combined with age-, sex-, and race-specific assumptions about mortality. Incidence rates were derived from the 1988 to 1999 Nationwide Inpatient Sample of the Healthcare Cost and Utilization Project, corrected for the likelihood of reamputation among those undergoing amputation for vascular disease. Incidence rates were assumed to remain constant over time and applied to historic mortality and population data along with the best available estimates of relative risk, future mortality, and future population projections. To investigate the sensitivity of our projections to increasing or decreasing incidence, we developed alternative sets of estimates of limb loss related to dysvascular conditions based on assumptions of a 10% or 25% increase or decrease in incidence of amputations for these conditions. Community, nonfederal, short-term hospitals in the United States. Persons who were discharged from a hospital with a procedure code for upper-limb or lower-limb amputation or diagnosis code of traumatic amputation. Not applicable. Prevalence of limb loss by age, sex, race, etiology, and level in 2005 and projections to the year 2050. In the year 2005, 1.6 million persons were living with the loss of a limb. Of these subjects, 42% were nonwhite and 38% had an amputation secondary to dysvascular disease with a comorbid diagnosis of diabetes mellitus. It is projected that the number of people living with the loss of a limb will more than double by the year 2050 to 3.6 million. If incidence rates secondary to dysvascular disease can be reduced by 10%, this number would be lowered by 225,000. One in 190 Americans is currently living with the loss of a limb. Unchecked, this number may double by the year 2050.


Characterization of 100 extended major histocompatibility complex haplotypes in Indonesian cynomolgus macaques.
Many medical advancements-including improvements to anti-rejection therapies in transplantation and vaccine development-rely on preclinical studies conducted in cynomolgus macaques (Macaca fascicularis). Major histocompatibility complex (MHC) class I and class II genes of cynomolgus macaques are orthologous to human leukocyte antigen complex (HLA) class I and class II genes, respectively. Both encode cell-surface proteins involved in cell recognition and rejection of non-host tissues. MHC class I and class II genes are highly polymorphic, so comprehensive genotyping requires the development of complete databases of allelic variants. Our group used PacBio circular consensus sequencing of full-length cDNA amplicons to characterize MHC class I and class II transcript sequences for a cohort of 293 Indonesian cynomolgus macaques (ICM) in a large, pedigreed breeding colony. These studies allowed us to expand the existing database of Macaca fascicularis (Mafa) alleles by identifying an additional 141 MHC class I and 61 class II transcript sequences. In addition, we defined co-segregating combinations of allelic variants as regional haplotypes for 70 Mafa-A, 78 Mafa-B, and 45 Mafa-DRB gene clusters. Finally, we defined class I and class II transcripts that are associated with 100 extended MHC haplotypes in this breeding colony by combining our genotyping analyses with short tandem repeat (STR) patterns across the MHC region. Our sequencing analyses and haplotype definitions improve the utility of these ICM for transplantation studies as well as infectious disease and vaccine research.


Magnetic resonance imaging in a family with hereditary cerebral arteriovenous malformations.
Because of a family history of neurologic problems and the documentation of three vascular lesions in one patient, we evaluated 18 members representing three family generations with magnetic resonance imaging. Of these, eight were normal, two had abnormalities probably not related to arteriovenous malformation, one scan was suboptimal, and the remaining eight had evidence of hemorrhagic lesions characteristic of arteriovenous malformation. Four of these patients had multiple lesions, and three patients with lesions had no neurologic symptoms. The findings suggest an autosomal dominant mode of inheritance in this unique case of familial cerebral arteriovenous malformation.


[Comparative study of the results of prosthesis implantation and reconstructive surgery of the mitral valve in congenital insufficiency].
In 23 of 154 patients with congenital mitral valve (MV) insufficiency the MV prosthetics was conducted, and in 23-valve-preserving reconstructive operation. Operative procedures were depicted, early and late follow-up results were analyzed. After the valve-preserving operation conduction hospital lethality was 8.7%, and after the (MV) prosthetics-34.8%. In the late follow-up period after (MV) plastic reconstruction conduction good results were noted in 17 patients, satisfactory-in 1, reoperation was performed in 3 (13%). Application of soft synthetic ring, preventing the dilation of the valve fibrotic ring, may to lower the frequency of reoperation conduction. Plastic correction of the CMVI permits to eliminate the basic haemodynamical disorders and to avoid or postpone the MV prosthetics conduction, what especially important in children.


Human immunodeficiency virus type 1 Rev- and Tat-specific cytotoxic T lymphocyte frequencies inversely correlate with rapid progression to AIDS.
Immunological correlates of AIDS-free survival after human immunodeficiency virus type 1 (HIV-1) infection are largely unknown. Cytotoxic T lymphocyte (CTL) responses are generally believed to be a major component of protective immunity against viral infections. However, the relationship between HIV-1-specific CTL responses and disease progression rate is presently unclear. Here we show in twelve HIV-1-infected individuals that detection of Rev-specific CTL precursors (CTLp) early in the asymptomatic stage, as well as detection of Rev- and Tat-specific CTLp later during follow-up, inversely correlate with rapid disease progression. No such correlation was found for detection of CTLp against Gag, RT or Nef. Further studies are required to determine whether a protective mechanism is indeed the basis of the observed correlation. The data presented are in agreement with the hypothesis that CTL against proteins that are important for early viral transcription and translation are of particular importance in protection from rapid disease progression.


Determination of biocatalytic parameters of a copper radical oxidase using real-time reaction progress monitoring.
An Auxiliary Activity Family 5 (AA5) copper-radical alcohol oxidase (AlcOx) with promiscuous activity towards simple alkyl and aromatic alcohols was evaluated using real-time reaction progress monitoring. Reaction kinetics using variable time normalization analysis (VTNA) were determined from reaction progress curves. By this approach, a detailed view of the entire reaction time course under various conditions was obtained and used to identify parameters that will inform further process optimization development. Optimal activity was found impacted by several factors, including reaction pH, oxygen saturation, and the source of a co-oxidant, either HRP or a chemical alternative, potassium ferricyanide. Analysis of reaction progress curves demonstrated that reaction stalling occurred as a result of oxygen depletion and from a loss of enzyme activity over time. The cooperativity between AlcOx, horseradish peroxidase (HRP), and catalase that result in enhanced reactivity was explored, with reaction pH being identified as a key factor for optimal activity. The results show that a process with HRP is more robust than with potassium ferricyanide, but that both oxidants likely activate AlcOx by a similar mechanism. The phenomenon of product inhibition was investigated for representative reactants, revealing that reaction inhibition was more significant for butyraldehyde than for benzaldehyde. Our analysis suggests that this is linked to the greater proportion in which butyraldehyde exists in the hydrated form.


Evidence for differential activation of arachidonic acid metabolism in formylpeptide- and macrophage-activation-factor-stimulated guinea-pig macrophages.
Alterations of phospholipid and arachidonic acid metabolism were studied by treatment of guinea-pig peritoneal-exudate macrophages with chemotactic peptide, formylmethionyl-leucylphenylalanine (fMet-Leu-Phe) and macrophage activation factor (MAF). The chemotactic peptide caused a rapid rearrangement in inositol phospholipids, including a breakdown of polyphosphoinositides within 30s, followed by a resultant formation of phosphatidylinositol (PI), diacylglycerol, phosphatidic acid and non-esterified arachidonic acid within 5 min. In addition to these sequential alterations, arachidonic acid was released mainly from PI. On the other hand, MAF induced a slow liberation of arachidonic acid, mainly from phosphatidylethanolamine (PE) and phosphatidylcholine (PC) by phospholipase A2 after the incubation period of 30 min, but not any rapid changes in phospholipids. Treatment of macrophages for 15 min with fMet-Leu-Phe produced the leukotrienes (LTs) B4, C4 and D4, prostaglandins (PG) E2 and F2 alpha and thromboxane (TX) B2. In contrast, MAF could not stimulate the production of arachidonic acid metabolites during the incubation period of 15 min, but could enhance that of PGE2, PGF2 alpha, TXB2 and hydroxyeicosatetraenoic acids at 6 h. However, the stimulated formation of LTs was not detected at any time. These results indicate that the effects of fMet-Leu-Phe on both phospholipid and arachidonic acid metabolism are very different from those mediated by MAF.


Transgenic overexpression of human DMPK accumulates into hypertrophic cardiomyopathy, myotonic myopathy and hypotension traits of myotonic dystrophy.
Abnormal expression of human myotonic dystrophy protein kinase (hDMPK) gene products has been implicated in myotonic dystrophy type 1 (DM1), yet the impact of distress accumulation produced by persistent overexpression of this poorly understood member of the Rho kinase-related protein kinase gene-family remains unknown. Here, in the aged transgenic murine line carrying approximately 25 extra copies of a complete hDMPK gene with all exons and an intact promoter region (Tg26-hDMPK), overexpression of mRNA and protein transgene products in cardiac, skeletal and smooth muscles resulted in deficient exercise endurance, an integrative index of muscle systems underperformance. In contrast to age-matched (11-15 months) wild-type controls, hearts from Tg26-hDMPK developed cardiomyopathic remodeling with myocardial hypertrophy, myocyte disarray and interstitial fibrosis. Hypertrophic cardiomyopathy was associated with a propensity for dysrhythmia and characterized by overt intracellular calcium overload promoting nuclear translocation of transcription factors responsible for maladaptive gene reprogramming. Skeletal muscles in distal limbs of Tg26-hDMPK showed myopathy with myotonic discharges coupled with deficit in sarcolemmal chloride channels, required regulators of hyperexcitability. Fiber degeneration in Tg26-hDMPK resulted in sarcomeric disorganization, centralization of nuclei and tubular aggregation. Moreover, the reduced blood pressure in Tg26-hDMPK indicated deficient arterial smooth muscle tone. Thus, the cumulative stress induced by permanent overexpression of hDMPK gene products translates into an increased risk for workload intolerance, hypertrophic cardiomyopathy with dysrhythmia, myotonic myopathy and hypotension, all distinctive muscle traits of DM1. Proper expression of hDMPK is, therefore, mandatory in supporting the integral balance among cytoarchitectural infrastructure, ion-homeostasis and viability control in various muscle cell types.


Risk factors for hantavirus infection in Germany, 2005.
In 2005, a marked increase in hantavirus infections was observed in Germany. Large cities and areas where hantaviruses were not known to be endemic were affected. A case-control study identified the following independent risk factors for infection: occupational exposure for construction workers, living <100 meter from forested areas, and exposure to mice.


[Methodologic studies in dynamic EMG mapping based on the Hilbert transformation].
On the basis of Discrete Hilbert Transformation spectral parameters for dynamic EMG analysis can be introduced. Within analysis intervals which are used in the same manner as in spectral analysis, the momentary power of EMG frequency bands can be computed as time series of the same interval duration. A close connection exists between momentary power and the current phenomenology of power spectral analysis because the momentary power of a defined frequency band can be seen as the exact dynamic equivalent of the corresponding mean power value deduced from the power spectrum. A multichannel EMG recording (greater than or equal to 16 channels) makes the representation of the topographical distribution of the spectral parameter by a coloured map possible. Additionally, the momentary power can be used to calculate map sequences. In this way, changes of EMG activity can be quantified by map sequences of an arbitrary time resolution. By calculation of momentary frequency via DHT an artefact detection scheme for ECG interference can be suggested. A completion of this methodology can be carried out by applications of adaptive filtration procedures. Using this concept of EMG processing, a new and common methodical basis of EMG power spectral analysis can be introduced.


Nonadiabatic dynamics at metal surfaces: independent electron surface hopping with phonon and electron thermostats.
Independent Electron Surface Hopping (IESH) is a computational method for accounting for nonadiabatic electronic transitions in simulations of molecular motion at metal surfaces. IESH is applicable in cases of strong coupling where the electronic friction model is suspect, and has been demonstrated to accurately reproduce the results of detailed molecular beam experiments on vibrationally inelastic scattering of nitric oxide from the (111) surface of gold. However, in its original form, IESH represents a closed system without energy flow outside the local region of explicitly included substrate atoms. In this paper we propose two "thermostats" for introducing energy flow to the bulk, one for phonons and the other for electronic excitations.


Microsatellite abnormalities in plasma of patients with breast carcinoma: concordance with the primary tumour.
To detect microsatellite abnormalities in the primary tumours and plasma of patients with breast carcinoma. Plasma was obtained from 17 breast carcinoma patients before surgery. Corresponding tumour and benign lymph node (control) samples for each of the carcinoma patients were obtained from paraffin blocks. DNA was extracted from the plasma samples and the paraffin embedded tissue using previously described methods. The 17 primary tumours showed two examples of loss of heterozygosity and three examples of microsatellite instability; the 17 plasma samples showed three and one, respectively. Many of the longer microsatellites (over 200 base pairs) were difficult to amplify from plasma. The investigations suggested that this was because of the highly fragmented nature of plasma DNA. Only one example of loss of heterozygosity and one example of microsatellite instability showed a concordant pattern in both primary tumour and plasma. These were both in the same patient. DNA mutations concordant with those in the primary carcinomas can occasionally be detected in the plasma of patients with breast carcinoma. However, the frequency would have to be markedly improved before this could be of any diagnostic value.


Psychological flexibility in coping with chronic pain: further examination of the brief pain coping inventory-2.
The role of coping in chronic pain management is well established. One challenge to the coping approach, however, is in identifying forms of coping that reliably lead to better functioning. An emerging approach to coping is based on the notion of psychological flexibility, a response pattern entailing openness to experience, awareness of specific behavioral options in a given situation, and persistence or alteration of activity according to personally held values and goals. A primary measure of psychological flexibility has been the Brief Pain Coping Inventory-2 (BPCI-2), and initial analyses have provided support for its utility in chronic pain treatment settings. The present study aimed to extend the previous work by examining relations of the BPCI-2 with measures of patient functioning, as well as with measures related to psychological flexibility, pain acceptance and valued activity in this case. A total of 324 individuals with chronic pain who completed a series of measures at an initial assessment appointment were included in the study. In correlation and regression analyses, the Psychological Flexibility subscale of the BPCI-2 achieved consistently significant relations with measures of disability, emotional functioning, pain acceptance, and valued activity, even after controlling for pain intensity and traditional coping methods. These results lend support to the adoption of psychological flexibility as a framework in future studies of coping with chronic pain.


Identifying gene expression changes in breast cancer that distinguish early and late relapse among uncured patients.
In recent years, microarray technology has revealed many tumor-expressed genes prognostic of clinical outcomes in early-stage breast cancer patients. However, in the presence of cured patients, evaluating gene effect on time to relapse is quite complex since it may affect either the probability of never experiencing a relapse (cure effect) or the time to relapse among the uncured patients (disease progression effect) or both. In this context, we propose a simple and an efficient method for identifying gene expression changes that characterize early and late recurrence for uncured patients. Simulation results show the good performance of the proposed statistic for detecting a disease progression effect. In a study of early-stage breast cancer, our results show that the proposed statistic provides a more powerful basis for gene selection than the classical Cox model-based statistic. From a biological perspective, many of the genes identified here as associated with the speed of disease recurrence have known roles in tumorigenesis.


[Character and risk factors of bacterial colonization of newborns in intensive care units].
A total of 116 newborns treated in intensive care wards were examined. The incidence, type, and risk factors of bacterial colonization were evaluated. Only 7.8% patients were colonized with gram-negative polyresistant microflora by admission, while after 3 days of intensive care as many as 48.2% newborns were colonized. The major microorganisms were Pseudomonas aeruginosa, Klebsiella spp., and Enterobacter spp. A relationship was detected between the bacterial spectrum of the skin and mucosa and the agents of infectious inflammatory complications in the newborns. The most significant factors of risk of colonization in the newborns with antibiotic-resistant gram-negative bacteria in intensive care wards were forced ventilation of the lungs and initially severe clinical status (CRIB score 5 and higher).


Marfan syndrome. Demonstration of abnormal elastin in aorta.
Aortae from three patients with classic presentation of Marfan syndrome, who died of vascular complications, were subjected to biochemical analyses of the connective tissue; for comparison, aortae from eight age-matched controls, without evidence of connective tissue abnormalities, were examined. Elastin was prepared from the aortae by two techniques. First, the tissues were extracted with 5 M guanidine-HCl, bacterial collagenase digestion and reduction with dithiothreitol (elastin I preparation). Secondly, this material was further purified by extraction with 0.1 M NaOH at 99 degrees C (elastin II preparation). Amino acid analyses of both elastin preparations indicated that the values for desmosine and isodesmosine were reduced in Marfan cases to approximately one-half of the control values. A corresponding increase in lysyl residues was noted in elastin II preparations. Also, the concentration of elastin per milligram dry weight of tissue was reduced in Marfan cases. The hydroxyproline content of elastin was increased in two cases with the Marfan syndrome. Recoveries indicated that the alkali treatment solubilized 46.2% of the elastin I preparations in Marfan aortae compared with 23.7% in controls. In contrast to elastin, the concentration and solubility of collagen were unchanged; the amino acid composition and the genetic types of insoluble collagen isolated by limited pepsin proteolysis were the same in both Marfan and control aortae. The results of our study demonstrate that the cross-linking of aortic elastin is reduced in the three patients with Marfan syndrome. Thus, a defect in elastin could explain the vascular fragility observed clinically in these patients.


Impact of mecA promoter mutations on mecA expression and beta-lactam resistance levels.
The reason for the extremely low-level oxacillin resistance in a so-called 'drug clone', a methicillin-resistant Staphylococcus aureus circulating among injection drug users in Zurich, Switzerland, could be traced back to the mecA promoter sequence and particularly to the strain's genetic background. Sequencing of its mec complex identified a point mutation (TATACT to TATATT), creating a perfect palindrome in the -10 region of the mecA promoter/operator region containing the binding sites for the mecA repressors MecI and BlaI. Two strains with vastly different beta-lactam resistance phenotypes, the low-level resistant drug clone type strain CHE482 and the highly homogeneously resistant strain COLn, were cured of their SCCmec elements and subsequently transformed with plasmids containing mecA under the control of either the wild-type or mutant promoter. Expression studies showed that this mutation had significant effects on both mecA transcription and corresponding PBP2a production, but only small effects on beta-lactam resistance levels within a given genetic background. A further mutation in the mecA ribosomal binding site (GGAGG to GGAGT), common to SCCmec type IV strains, was found to have no discernable effect on mecA transcription and PBP2a content, and only minimal effects on beta-lactam resistance. Factors associated with the genetic backgrounds into which these differently controlled mecA genes were introduced had a much higher impact on beta-lactam resistance levels than the rates of mecA transcription. The tight repression of mecA expression in this drug clone in the absence of beta-lactams could contribute to the apparent fitness of this fast growing strain.