Implantation of recombinant rat myocytes into adult skeletal muscle: a potential gene therapy.
The ability of skeletal muscle to regenerate provides an excellent therapeutic entry, via genetic engineering, for correcting diseases of skeletal muscle and other tissues. We have used a retrovirus to transfer the cDNA for the human multidrug transporter, encoded by the MDR1 gene, into the genomes of the rat muscle cell line L6 and into primary rat myocytes. The MDR1 gene confers drug resistance to cells, and thus serves as a selectable marker in vitro. In cultured cells, the retroviral promoter-driven human MDR1 cDNA was shown to be stable in the presence or absence of drug selection or muscle cell fusion. MDR1 mRNA was synthesized, as shown by RNA blot analysis and in situ hybridization. The protein product was localized to the plasma membrane of transduced myocytes and myotubes by immunofluorescence. As a model for skeletal muscle gene therapy, transduced L6 myocytes were implanted into the tibialis anterior muscle of Wistar rats. The retroviral sequences of the human MDR1 gene and its mRNA were present in the muscles of Wistar rats 5 days, but not 12 days, after implantation, possibly because of immunorejection. On the other hand, the human MDR1 cDNA was stable in the tibialis anterior muscle of nude mice, which are incapable of immunorejection, at least 4 weeks after implantation of myocytes. Immunosuppression of Wistar rats with cyclosporine A delayed immunorejection of recombinant myocytes, and MDR1 cDNA and mRNA was detected 3-4 weeks after implantation. In situ hybridization revealed that injected recombinant myocytes remain in discrete foci in adult rodent skeletal muscle and express MDR1 mRNA for at least 30 days in nude mice and cyclosporine-treated rats.


Social network resources and management of hypertension.
Hypertension is one of the most prevalent chronic diseases among older adults, but rates of blood pressure control are low. In this article, we explore the role of social network ties and network-based resources (e.g., information and support) in hypertension diagnosis and management. We use data from the National Social Life, Health, and Aging Project to identify older adults with undiagnosed or uncontrolled hypertension. We find that network characteristics and emotional support are associated with hypertension diagnosis and control. Importantly, the risks of undiagnosed and uncontrolled hypertension are lower among those with larger social networks-if they discuss health issues with their network members. When these lines of communication are closed, network size is associated with greater risk for undiagnosed and uncontrolled hypertension. Health care utilization partially mediates associations with diagnosis, but the benefits of network resources for hypertension control do not seem to stem from health-related behaviors.


Reversible guest exchange and ferrimagnetism (T(C) = 60.5 K) in a porous cobalt(II)-hydroxide layer structure pillared with trans-1,4-cyclohexanedicarboxylate.
The synthesis, characterization, and reversible guest-exchange chemistry of a new porous magnetic material that orders ferrimagnetically at 60.5 K are described. The material, Co(5)(OH)(8)(chdc).4H(2)O (chdc = trans-1,4-cyclohexanedicarboxylate), contains tetrahedral-octahedral-tetrahedral Co(II)-hydroxide layers of composition Co((oct))(3)Co((tet))(2)(OH)(8) that are linked together by bis(unidentate) chdc pillars. Noncoordinated water molecules occupy 1-D channels situated between the chdc pillars. The material remains monocrystalline during dehydration from Co(5)(OH)(8)(chdc).4H(2)O (CDCC.4H(2)O) to Co(5)(OH)(8)(chdc) (CDCC) via an intermediate Co(5)(OH)(8)(chdc).2H(2)O (CDCC.2H(2)O) upon heating or evacuation. In-situ single crystal and powder X-ray diffraction analyses indicate that the interlayer spacing decreases in two steps, each corresponding to the loss of two water molecules per formula unit as determined by thermogravimetry. The single crystal structure of the fully dehydrated material, CDCC, has no void volume due to a tilting of the pillars and 9% decrease of the interlayer spacing with water removal. Exposure of CDCC to air causes rapid rehydration of this material to CDCC.4H(2)O, as determined by single crystal X-ray diffraction, powder X-ray diffraction, thermogravimetry, and vibrational spectroscopy. Both the hydrated and dehydrated forms order magnetically below 60.5 K. The susceptibility data are consistent with ferrimagnetic behavior, and the value of the saturation magnetization at 2 K (ca. 2 micro(B)) is explained by a model of two sublattices, one comprising three octahedral cobalt atoms and another comprising two tetrahedral cobalt atoms. There is an enhanced 2-D correlation within the layer at temperatures just above the Curie temperature, as seen by nonlinearity in the ac susceptibility data and remanence in the isothermal magnetization. The crossover from 2-D to 3-D ordering occurs at T(C). The large anisotropy in the magnetization data on a single crystal suggests either a 2-D Ising or an XY magnet while the critical exponent of 0.25 is in favor of the latter. Both magnetization data in a small field in the ac and dc mode and isothermal magnetization data provide evidence of a further change in behavior at 23 K, which may originate from a reorientation of the moments within the layer. Variation of the pillar and of the guest-exchange chemistry, including the exchange of magnetic guests such as O(2), offers the possibility of tailoring the magnetic properties of this material.


A qualitative study of children's snack food packaging perceptions and preferences.
Food marketing is pervasive in high- and low/middle-income countries and is recognized as a significant risk factor for childhood obesity. Although food packaging is one of the most important marketing tools to persuade consumers at the point-of-sale, scant research has examined how it influences children's perceptions. This study was conducted in Guatemala and aimed to understand which snack foods are the most frequently purchased by children and how aspects of food packaging influence their product perceptions. Six activity-based focus groups were conducted in two elementary public schools with thirty-seven children (Grades 1 through 6, age range 7-12 years old). During each focus group, children participated in three activities: 1) list their most frequently purchased food products; 2) select the picture of their favorite product, the packaging they liked best, and the product they thought was the healthiest from eight choices; and 3) draw the package of a new snack. Children reported purchasing salty snacks most frequently. Most children chose their favorite product based on taste perceptions, which can be influenced by food packaging. Visual elements influenced children's selection of favorite packaging (i.e., characters, colors) and healthiest product (i.e., images), and persuaded some children to incorrectly think certain foods contained healthy ingredients. When children generated their own drawings of a new product, the most frequently included packaging elements in the drawings were product name, price, product image and characters, suggesting those aspects of the food packaging were most significant to them. Policies regulating package content and design are required to discourage consumption of unhealthy snacks. This might be another public health strategy that can aid to halt the obesity epidemic.


Secretin-Enhanced Magnetic Resonance Cholangiopancreatography for Assessing Pancreatic Secretory Function in Children.
To assess the accuracy and interrater reproducibility of measurements of pancreatic secretory function by magnetic resonance cholangiopancreatography in response to secretin administration and to describe our experience using the technique to noninvasively assess pancreatic secretory function in a pediatric population. In the accuracy study, phantoms with varying fluid volume (47-206 mL) were imaged using the clinical quantification sequence. Fluid volume was measured by image segmentation (ImageJ). Measurement accuracy was expressed in terms of error (absolute and percent) relative to known fluid volume. In the reproducibility study and clinical experience, 31 patients with suspected pancreatic disease underwent 33 secretin-enhanced magnetic resonance cholangiopancreatography exams. Two-dimensional T2-weighted, fat-saturated single shot fast spin echo sequences were acquired before and after secretin injection (0.2 µg/kg, max 16 µg). Secreted fluid volume (postsecretin minus presecretin) was independently measured by 2 blinded reviewers. Between reviewer measurement reproducibility was assessed based on correlation (Spearman) and bias (Bland-Altman analysis). For the accuracy study, fluid volumes were measured with mean volume errors of -0.3 to +12.5 mL (percent error -0.03% to +9.0%). For the reproducibility study, the mean secreted fluid volumes measured by reviewer 1 and reviewer 2 were 79.1 ± 54.3 mL (range 5.5-215.4) and 77.2 ± 47.1 mL (range 6.7-198.1 mL), respectively. Measured secreted fluid volumes were very strongly correlated (r = 0.922) between reviewers with a bias of only 1.9 mL (95% limits of agreement -40.5 to 44.2). Measurement of fluid volume by magnetic resonance imaging is highly accurate with <10% (<13 mL) error in measured volume. Measurements of pancreatic secreted fluid volume in response to secretin by magnetic resonance cholangiopancreatography are highly reproducible with a bias of <2 mL between reviewers.


Magnetic field fluctuation measurement with a heavy ion beam probe in the Large Helical Device.
The measurements of magnetic field fluctuation with a heavy ion beam probe (HIBP) in the Large Helical Device (LHD) are considered in this article. Measured value reflects not only the fluctuation at the observation point (sample volume) but also the fluctuation on the beam path. These effects (called as the path integral effects) are considered and calculated for variations of fluctuations in LHD. The distortion of the measured profile by the path integral effect is not significant at the peak of the fluctuation amplitude. In contrast, in the region far from the peak, phony signals due to path integral effects are observed. Finally, the resolution of magnetic field fluctuation measurements with our HIBP system is estimated. Estimated results show that the magnitude of 4.1x10(-5) T can be measured with our system.


Combined dynamic light scattering and Raman spectroscopy approach for characterizing the aggregation of therapeutic proteins.
Determination of the physicochemical properties of protein therapeutics and their aggregates is critical for developing formulations that enhance product efficacy, stability, safety and manufacturability. Analytical challenges are compounded for materials: (1) that are formulated at high concentration, (2) that are formulated with a variety of excipients, and (3) that are available only in small volumes. In this article, a new instrument is described that measures protein secondary and tertiary structure, as well as molecular size, over a range of concentrations and formulation conditions of low volume samples. Specifically, characterization of colloidal and conformational stability is obtained through a combination of two well-established analytical techniques: dynamic light scattering (DLS) and Raman spectroscopy, respectively. As the data for these two analytical modalities are collected on the same sample at the same time, the technique enables direct correlation between them, in addition to the more straightforward benefit of minimizing sample usage by providing multiple analytical measurements on the same aliquot non-destructively. The ability to differentiate between unfolding and aggregation that the combination of these techniques provides enables insights into underlying protein aggregation mechanisms. The article will report on mechanistic insights for aggregation that have been obtained from the application of this technique to the characterization of lysozyme, which was evaluated as a function of concentration and pH.


Chronic shoulder dislocations.
Thirty-two patients with chronic dislocation of the shoulder were diagnosed over a 5-year period at King Edward VIII Hospital, Durban, South Africa. Thirty-one patients had an anterior subcoracoid dislocation. One patient had a chronic posterior dislocation. The duration of dislocation ranged from 6 days to 2 years. Six patients were epileptic, and it was an affliction of all ages. None of the patients had a vascular deficit, but 4 had neurologic damage on presentation. Closed manipulation was only successful in 1 patient. Ten patients' shoulders were left unreduced in the dislocated position. Twenty-one patients, including the patient with a single posterior dislocation, underwent open reduction. The author treated 10 patients with chronic anterior dislocations surgically. The coracoid was transferred to the glenoid, and acromiohumeral K-wire fixation was used for 4 weeks in this group. These 10 cases, as well as 5 cases left unreduced, were followed up for more than 2 years. The results were evaluated by the system proposed by Rowe and Zarins and showed that surgically treated patients fared better than those whose shoulders were left unreduced regardless of the duration of the dislocation or the age of the patient. This series differs from previous reports in several respects. All but one of the dislocations were anterior, the incidence of epilepsy was lower, and successful reduction by manipulation was rarely achieved. In all 10 cases in which the author performed the operation, the shoulder was successfully relocated without neurovascular complications. Dissection of the axillary vessels and nerves was never necessary.


[Sonographic localization of parathyroid adenomas in hyperparathyroidism].
The diagnostic efficacy of sonography for localisation of adenomas of the parathyroid glands in the neck region was investigated in 13 patients with hypercalcaemia and increased serum parathormone levels. Solitary parathyroid adenoma was demonstrated in all patients. The diagnosis was confirmed by surgery in 12 patients. In one patient, adenoma of the parathyroid can be assumed due to sonographically demonstrated increase in size and increase of serum parathormone. Adenomas of the parathyroid are characterised sonographically by typical dorsal position at the posterior surface of the thyroid, by echo-paucity and echo-density of the "capsular structure" around the adenomas. Sonography is suitable as screening method in hypercalcaemia and in increased parathormone serum levels for preoperative localisation of parathyroid adenomas.


Mutations affecting the pattern of the larval cuticle inDrosophila melanogaster : II. Zygotic loci on the third chromosome.
The present report describes the recovery and genetic characterization of mutant alleles at zygotic loci on the third chromosome ofDrosophila melanogaster which alter the morphology of the larval cuticle. We derived 12600 single lines from ethyl methane sulfonate (EMS)-treatedst e orrucuca chromosomes and assayed them for embryonic lethal mutations by estimating hatch rates of egg collections. About 7100 of these lines yielded at least a quarter of unhatched eggs and were then scored for embryonic phenotypes. Through microscopic examination of unhatched eggs 1772 lines corresponding to 24% of all lethal hits were classified as embryonic lethal. In 198 lines (2.7% of all lethal hits), mutant embryos showed distinct abnormalities of the larval cuticle. These embryonic visible mutants define 45 loci by complementation analysis. For 32 loci, more than one mutant allele was recovered, with an average of 5.8 alleles per locus. Complementation of all other mutants was shown by 13 mutants. The genes were localized on the genetic map by recombination analysis, as well as cytologically by complementation analysis with deficiencies. They appear to be randomly distributed along the chromosome. Allele frequencies and comparisons with deficiency phenotypes indicate that the 45 loci represent most, if not all, zygotic loci on the third chromosome, where lack of function recognizably affects the morphology of the larval cuticle.


The abortion pill: a solution for unsafe abortions in developing countries?
The abortion pill is a new abortion technique which does not require technically skilled health personnel. Several clinical researchers have stated that the ease of administration of the abortion pill gives it the potential to save lives in countries where thousands of women die annually from complications of failed abortions due to poor health services. This article discusses medical and users' aspects of the abortion pill, with particular emphasis on its use in developing countries, and questions the usefulness of the abortion pill in areas where health care facilities are in short supply. It stresses the need to consider the social, cultural and health care context in which the abortion pill will be used before it is sold on the world market. The safety and efficacy of the abortion pill could be adversely affected by the way women perceive its effectiveness, women's beliefs about conception and pregnancy, and their health status. In the present two-phase administration form the abortion pill is not likely to be appropriate for use in developing countries with a shortage of health care facilities.


Silencing of HIV-1 gene expression by two types of siRNA expression systems.
The RNA interference (RNAi) phenomenon is a recently discovered process in which the introduction of a double-stranded RNA (dsRNA) into cells causes the specific degradation of mRNA containing the same sequence. We designed mammalian expression vectors that direct the synthesis of small interfering RNA (siRNA)-like transcripts and examined them for their siRNA-mediated gene interference targeting the env gene (NL4-3:7490-7508, E7490). We constructed siRNA expression vectors for two different strands (sense and antisense; tandem promoter) and for siRNA expressed from the short hairpin RNA (shRNA). The inhibition efficacy on HIV-1 replication differed between these two vectors. Notably, the shRNA vector pU6-env-shRNA inhibited p24 production more effectively than the tandem promoter expression vector pU6-env-siRNA. Furthermore, we examined the ability of lentiviral vectors expressing shRNA to suppress HIV-1 expression in HIV-1-infected SupT1 cells. The env-shRNA (E 7490) almost completely suppressed HIV-1 expression in infected cells for up to 15 days.


Signal transduction in vertebrate growth cones navigating in vivo.
Navigating growth cones need signal transduction machinery to amplify and transmit the effects of extracellular signals throughout the growth cone. In culture, many drugs that affect second messengers are known to modulate neurite extension (with different effects on different neurons), and gradients of calcium influx and cyclic nucleotide analogs can cause growth cones to turn. However, it is not clear which of these responses are physiologically relevant, as axons grow through much more complex environments in vivo. The "exposed brain" preparation in Xenopus embryos provides an experimentally tractable system in which it is possible to study growth, pathfinding, and target recognition of retinal growth cones in vivo, while pharmacologically manipulating their signal transduction systems. These growth cones can also be easily studied in explant culture. We describe preliminary results of parallel in vivo and in vitro experiments using an array of drugs that perturb transduction molecules. Surprisingly, calcium ionophores and cyclic nucleotide analogs have no significant effect on retinal axon growth or pathfinding. Several agents including herbimycin A, ML-7, mastoparan, and RHC80267 inhibit retinal axon growth, both in vivo and in vitro, suggesting that tyrosine kinases, myosin, heterotrimeric G-proteins, and diacylglycerol lipase are important for retinal growth cones navigating in the optic pathway.


Hormonal regulation of the annual pelage color cycle in the Djungarian hamster, Phodopus sungorus. I. Role of the gonads and pituitary.
The Djungarian hamster exhibits an agouti pelage in the summer and a predominantly white pelage in the winter. This pelage color cycle is known to be regulated by the length of the daily photoperiod probably acting through the pineal gland, as is the seasonal cycle of reproductive function with which it is closely correlated ( Figala et al., '73; Hoffmann, ' 78b ). The possibility of a causal relationship between the decline in gonadal hormone secretion and the coat color change occurring in short photoperiod was examined. Gonadectomized and intact male and female hamsters were exposed to either long (16L:8D) or short ( 10L : 14D ) photoperiod for several months. Gonadectomy neither induced the change to the winter pelage color in long photoperiod-housed animals, nor prevented either the change to the winter pelage or the spontaneous return to summer pelage color in short photoperiod-housed animals. Chronic implants of testosterone in castrated males delayed and attenuated the short photoperiod-induced coat color change. Administration of ovine prolactin (100 micrograms/day) stimulated pigmentation in hamsters with the winter pelage, whereas administration of a alpha MSH (30 micrograms/day) was without effect. These results suggest that changes in pelage color may be regulated largely by changes in pituitary prolactin secretion and modified to some extent by changes in gonadal steroid hormone secretion.


Embryonic even skipped-dependent muscle and heart cell fates are required for normal adult activity, heart function, and lifespan.
The Drosophila pair-rule gene even skipped (eve) is required for embryonic segmentation and later in specific cell lineages in both the nervous system and the mesoderm. We previously generated eve mesoderm-specific mutants by combining an eve null mutant with a rescuing transgene that includes the entire locus, but with the mesodermal enhancer removed. This allowed us to analyze in detail the defects that result from a precisely targeted elimination of mesodermal eve expression in the context of an otherwise normal embryo. Absence of mesodermal eve causes a highly selective loss of the entire eve-expressing lineage in this germ layer, including those progeny that do not continue to express eve, suggesting that mesodermal eve precursor specification is not implemented. Despite the resulting absence of a subset of muscles and pericardial cells, mesoderm-specific eve mutants survive to fertile adulthood, providing an opportunity to examine the effects of these developmental abnormalities on adult fitness and heart function. We find that in these mutants, flying ability, myocardial performance under normal and stressed conditions, and lifespan are severely reduced. These data imply a nonautonomous role of the affected pericardial cells and body wall muscles in developing and/or maintaining cardiac performance and possibly other functions contributing to normal lifespan. Given the similarities of molecular-genetic control between Drosophila and vertebrates, these findings suggest that peri/epicardial influences may well be important for proper myocardial function.


1,3-Dipolar Cycloadditions of 4-Acetoxy Allenoates: Access to 2,3-Dihydropyrazoles, 2,3-Dihydroisoxazoles, and Indolizines.
The thermal 1,3-dipolar cycloadditions of 4-acetoxyallenoates 1 with various dipoles have been reported. When azomethine imines and nitrones are used as the 1,3-dipole partner, the corresponding reactions afford 2,3-dihydropyrazole and 2,3-dihydroisoxazole derivatives, respectively. These reactions might proceed via a thermal 1,3-dipolar cycloaddition and the subsequent elimination of HOAc. In addition, allenoates 1 react well with in situ generated azomethine ylides in which a similar cycloaddition pathway is followed by oxidative aromatization to give indolizine derivatives.


Understanding how a sport-helmet protects the head from closed injury by virtual impact tests.
Understanding how a helmet protects the head, especially the soft brain tissues, is the prerequisite for improving helmet design. Intracranial pressure and stresses/strains in the brain tissues are the direct indicators of traumatic brain injury and they can be used to measure helmet performance. In this study, the effects of helmet design parameters such as the helmet shell stiffness, liner compliance and thickness on the brain injury indicators were investigated by virtual impact tests. A finite element head model (FEHM) was first constructed from medical images; a personally-fitted helmet made of composite material and foam was virtually prototyped using geometric information extracted from the FEHM; a helmet-head finite element model was then assembled. Virtual impact tests were conducted using the resulting helmet-head model. The obtained results suggested that, if the helmet shell already has adequate strength to resist excessive deformation and fracture, further increasing shell stiffness and strength would not considerably reduce intracranial pressure and brain strains; to reach the maximum protection with the available materials, the key is to effectively use the second stage in the stress-strain history of the liner foam material.


Antimicrobial potentiality of a new non-antibiotic: the cardiovascular drug oxyfedrine hydrochloride.
Ten cardiovascular drugs, having diverse pharmacological action, were screened for possible antimicrobial property against known eight sensitive bacteria, belonging to Gram positive and Gram negative types. Although five drugs failed to show antimicrobial activity and three had moderate antimicrobial action, oxyfedrine HCl and dobutamine were seen to possess pronounced antimicrobial property. Oxyfedrine was further tested in vitro against 471 strains of bacteria from two Gram positive and fourteen Gram negative genera. The minimum inhibitory concentration (MIC) of oxyfedrine was determined by agar dilution method, which ranged from 50-200 microg/ml in most of the strains, while some strains were inhibited at even lower concentrations. In animal experiments, this compound was capable of offering significant protection to Swiss strain of white mice, challenged with 50 median lethal dose (MLD) of a virulent strain of Salmonella typhimurium at concentrations of 15, 30 and 60 microg/mouse. The in vivo results were highly significant according to chi-square test.


Variant translocation partners of the anaplastic lymphoma kinase (ALK) gene in two cases of anaplastic large cell lymphoma, identified by inverse cDNA polymerase chain reaction.
In anaplastic large cell lymphoma (ALCL), the anaplastic lymphoma kinase (ALK) gene is rearranged with diverse partners due to variant translocations/inversions. Case 1 was a 39-year-old man who developed multiple tumors in the mediastinum, psoas muscle, lung, and lymph nodes. A biopsy specimen of the inguinal node was effaced by large tumor cells expressing CD30, epithelial membrane antigen, and cytoplasmic ALK, which led to a diagnosis of ALK(+) ALCL. Case 2 was a 51-year-old man who was initially diagnosed with undifferentiated carcinoma. He developed multiple skin tumors eight years after his initial presentation, and was finally diagnosed with ALK(+) ALCL. He died of therapy-related acute myeloid leukemia. G-banding and fluorescence in situ hybridization using an ALK break-apart probe revealed the rearrangement of ALK and suggested variant translocation in both cases. We applied an inverse cDNA polymerase chain reaction (PCR) strategy to identify the partner of ALK. Nucleotide sequencing of the PCR products and a database search revealed that the sequences of ATIC in case 1 and TRAF1 in case 2 appeared to follow those of ALK. We subsequently confirmed ATIC-ALK and TRAF1-ALK fusions by reverse transcriptase PCR and nucleotide sequencing. We successfully determined the partner gene of ALK in two cases of ALK(+) ALCL. ATIC is the second most common partner of variant ALK rearrangements, while the TRAF1-ALK fusion gene was first reported in 2013, and this is the second reported case of ALK(+) ALCL carrying TRAF1-ALK.


Widespread changes in nucleosome accessibility without changes in nucleosome occupancy during a rapid transcriptional induction.
Activation of transcription requires alteration of chromatin by complexes that increase the accessibility of nucleosomal DNA. Removing nucleosomes from regulatory sequences has been proposed to play a significant role in activation. We tested whether changes in nucleosome occupancy occurred on the set of genes that is activated by the unfolded protein response (UPR). We observed no decrease in occupancy on most promoters, gene bodies, and enhancers. Instead, there was an increase in the accessibility of nucleosomes, as measured by micrococcal nuclease (MNase) digestion and ATAC-seq (assay for transposase-accessible chromatin [ATAC] using sequencing), that did not result from removal of the nucleosome. Thus, changes in nucleosome accessibility predominate over changes in nucleosome occupancy during rapid transcriptional induction during the UPR.


Designed multiple ligands for the treatment of type 2 diabetes mellitus and its complications: Discovery of (5-arylidene-4-oxo-2-thioxothiazolidin-3-yl)alkanoic acids active as novel dual-targeted PTP1B/AKR1B1 inhibitors.
Type 2 diabetes mellitus (T2DM) is a serious chronic disease with an alarmingly growing worldwide prevalence. Current treatment of T2DM mainly relies on drug combinations in order to control blood glucose levels and consequently prevent the onset of hyperglycaemia-related complications. The development of multiple-targeted drugs recently emerged as an attractive alternative to drug combinations for the treatment of complex diseases with multifactorial pathogenesis, such as T2DM. Protein tyrosine phosphatase 1B (PTP1B) and aldose reductase (AKR1B1) are two enzymes crucially involved in the development of T2DM and its chronic complications and, therefore, dual inhibitors targeted to both these enzymes could provide novel agents for the treatment of this complex pathological condition. In continuing our search for dual-targeted PTP1B/AKR1B1 inhibitors, we designed new (5-arylidene-4-oxo-2-thioxothiazolidin-3-yl)alkanoic acids. Among them, 3-(4-phenylbutoxy)benzylidene derivatives 6f and 7f, endowed with interesting inhibitory activity against both targets, proved to control specific cellular pathways implicated in the development of T2DM and related complications.


Volatile flavor constituents in the pork broth of black-pig.
Pork of black-pig in China is well known for its quality and preferred by consumers. However, there is a lack of research on its flavors. By solvent assisted flavor evaporation combined with GC-MS, 104 volatile compounds in the stewed pork broth of black-pig were identified with the dominant amounts of fatty acids, alcohols, and esters. By aroma extract dilution analysis-GC-O method, 27 odor-active compounds were characterized, including 2-methyl-3-furanthiol, 3-(methylthio)propanal, 2-furfurylthiol, γ-decalactone, nonanal, (E)-2-nonenal, and (E,E)-2,4-decadienal that had high FD factors. Compared to the common white-pig, the aroma compounds in both pork broths were almost the same, but the aroma profile of potent odorants for the black-pig pork broth showed less fatty and more roasted notes, which were partially attributed to the higher monounsaturated fatty acids and lower polyunsaturated fatty acids in meat. With aid of authentic chemicals and selected reaction monitoring mode of GC-MS/MS, 19 aroma compounds were quantitated.


Safety assessment of bacterial choline oxidase protein introduced in transgenic crops for tolerance against abiotic stress.
Genetically modified crops have resistance to abiotic stress by introduction of choline oxidase protein. In the present study, the safety of choline oxidase protein derived from Arthrobacter globiformis was assessed for toxicity and allergenicity. The protein was stable at 90 degrees C for 1 h. Toxicity studies of choline oxidase in mice showed no significant difference (p > 0.05) from control in terms of growth, body weight, food consumption, and blood biochemical indices. Histology of gut tissue of mice fed protein showed normal gastric mucosal lining and villi in jejunum and ileum sections. Specific IgE in serum and IL-4 release in splenic culture supernatant were low in choline oxidase treated mice, comparable to control. Intravenous challenge with choline oxidase did not induce any adverse reaction, unlike ovalbumin group mice. Histology of lung tissues from choline oxidase sensitized mice showed normal airways, whereas ovalbumin-sensitized mice showed inflamed airways with eosinophilic infiltration and bronchoconstriction. ELISA carried out with food allergic patients' sera revealed no significant IgE affinity with choline oxidase. Also, choline oxidase did not show any symptoms of toxicity and allergenicity in mice.


Deletion 2q37 syndrome: Cognitive-behavioral trajectories and autistic features related to breakpoint and deletion size.
Subtelomeric deletions have been reported in ∼2.5% of individuals with developmental disabilities. Subtelomeric deletion 2q37 has been detected in many individuals diagnosed with intellectual disabilities (ID) and autism spectrum disorders (ASD). Previously, genotype-phenotype correspondences were examined for their relationship to breakpoints 37.1, 37.2, or 37.3. Our purpose was to ascertain whether there were phenotypic differences at these breakpoints, elucidate the cognitive-behavioral phenotype in del2q37, and examine the genotype-phenotype association in the deletion with respect to cognitive-behavioral profiles and ASD. We administered a comprehensive cognitive-behavioral battery to nine children diagnosed with del 2q37, ages 3.9-17.75 years. ID for five tested with the Stanford-Binet (4th Edition) (SBFE) ranged from severe to mild [IQ Range: 36-59]. Adaptive behavior scores from the Vineland Adaptive Behavior Scale (VABS) were much below adequate levels (DQ Range: floor value ["19"] to 55). Autism scores from the Child Autism Rating Scale (CARS) ranged from 22 [non-autistic] to 56 [extremely autistic]; 5/8 [63%] children received scores on the autism spectrum. Participants with the largest deletions, 10.1 and 9.5 Mb, attained the highest IQ and DQ scores while those with the smallest deletions, 7.9 and 6.6 Mb, made the lowest IQ and DQ scores. No association between deletion breakpoint and phenotype were found. Assessment of the various deleted regions suggested histone deacetylase 4 gene (HDAC4) was a likely candidate gene for ASD in our sample. However, two earlier reports found no association between HDAC4 haploinsufficiency and ASD. © 2016 Wiley Periodicals, Inc.


[Prevention of early and late complications after gastrectomy].
A procedure for making esophago-intestinal anastomosis was experimentally developed and integrated with clinical practice. The method consists in formation of a muscular constrictor in the distal part of the gullet and a mucosa valve. Surgery was performed in 52 patients: gastric cancer--50 (stage II--3, III--44 and IV--3) and subcardial ulcer--2 cases. Extended combined gastrectomy was used in 14 patients (27.0%). Neither anastomotic failure nor lethality was observed. Late-onset 2. reflux-esophagitis was diagnosed in 2 out of 39 (5.1%) endoscopically examined patients.


Evaluation of breastfeeding promotion, support, and knowledge of benefits on breastfeeding outcomes.
We examined how prenatal exposure to breastfeeding information from various media sources, maternal knowledge of benefits, family and clinician support, and peer practices influence breastfeeding outcomes in early infancy. Initiation of breastfeeding, any breastfeeding at two months, and exclusivity of breastfeeding at two months were examined in a cohort of US women using data from the Infant Feeding Practices Study II. Descriptive statistics, chi-square analyses and logistic regression were conducted. Approximately 85 percent of the women initiated breastfeeding. At two months, 63.8 percent continued breastfeeding, while only 38.1 percent breastfed exclusively. Mothers with greater knowledge about breastfeeding benefits were 11.2 (95% CI: 6.87-18.45) times more likely to initiate breastfeeding and 5.62 (95% CI: 4.19-7.54) times more likely to breastfeed at two months than those with lower levels of knowledge. Women whose families prenatally supported exclusive breastfeeding were 8.21(5.12-13.2) times more likely to initiate and continue breastfeeding (OR 3.21, 95% CI: 2.51-4.11). Clinicians who supported breastfeeding only also increased the odds of a woman initiating breastfeeding (OR 1.95, 95% CI: 1.31-2.88). Interventions to increase maternal knowledge of breastfeeding benefits and family and clinician support of breastfeeding in the prenatal period may help increase breastfeeding rates. The encouragement of breastfeeding needs to be a priority among health care providers to improve the health of mothers and infants.


Content of metals in rabbit tissues.
The purpose of the experiments was the determination of lead, cadmium, iron, copper, zinc, manganese and magnesium in some internal organs and muscles of rabbits. The investigations were performed in 1990 with 360 white New Zealand and black-white rabbits which were slaughtered when they had 2.0 kg in weight. The experimental material (180 animals) came from a rabbit farm kept by Department of Genetics and Animal Breeding in the Agricultural Academy of Cracow. The control group constituted 180 rabbits from an area without the industry. With regard to the average lead content in 22% of kidneys and liver of experimental rabbit group an exceeding of the allowable limit and in average cadmium content in 18% of kidneys and liver was stated. The cadmium contents of values for the kidneys, liver, heart and muscles are several times higher compared with the highest values which were determined for the organs and muscles from the control area.


A Case of Wellens Syndrome in a 30-Year-Old Woman From Sub-Saharan Africa: A Perplexing Clinical Entity With Invaluable Lessons.
With an estimated contribution of up to 6% of all acute coronary events, young adults are experiencing an escalating burden and mortality attributable to coronary artery disease (CAD) worldwide. Wellens syndrome, a preinfarction clinical entity with distinctive electrocardiographic (ECG) features and high propensity for extensive anterior wall infarctions, affects about 15% of unstable angina subpopulation. We report challenges and lessons learnt from the first ever documented case of Wellens syndrome in Tanzania. A 30-year-old female of African descent was referred to us from an upcountry zonal referral hospital for etiological determination of chest pain and expert management. Her medical history was unremarkable, and she had no apparent risk factors for CAD. She presented with a 7-day history of ongoing sharp central chest pain that was radiating to the neck and jaws and relieved momentarily by morphine. She had stable vitals with an unremarkable systemic examination; however, a 12-lead ECG revealed deeply inverted T-waves on leads V2 through V4. Based on our extensive history and physical examination we came up with a diagnosis of type B Wellens syndrome with impending anterior wall myocardial infarction. She underwent cardiac catheterization that revealed a nonthrombogenic severe subocclusive (>95%) mid left anterior descending artery stenosis. Subsequently, angioplasty was performed successfully with a resolute integrity stent, and TIMI III flow was achieved. To conclude, despite its relative frequency, physicians' awareness of Wellens syndrome pathognomonic ECG features is of paramount importance to curb its attributable morbidity and mortality.


Surface Modification in CsPb0.5Sn0.5I2Br Inorganic Perovskite Solar Cells: Effects of Bifunctional Dipolar Molecules on Photovoltaic Performance.
Inorganic tin-lead binary perovskites have piqued the interest of researchers as effective absorbers for thermally stable solar cells. However, the nonradiative recombination originating from the surface undercoordinated Sn2+ cations and the energetic offsets between different layers cause an excessive energy loss and deteriorate the perovskite device's performance. In this study, we investigated two thioamide derivatives that differ only in the polar part connected to their common benzene ring, namely, benzenecarbothioamide and 4-fluorophenylcarbothioamide (F-TBA). These two molecules were implemented as modifiers onto the inorganic tin-lead perovskite (CsPb0.5Sn0.5I2Br) surface in the perovskite solar cells. Modifiers that carry C═S and NH2 functional groups, equipped with lone electron pairs, can autonomously associate with surface Sn2+ through coordination and electrostatic attraction mechanisms. This interaction serves effectively to passivate the surface. In addition, due to the permanent dipole moment of the intermediate layer, an interfacial dipole field appears at the PCBM/CsPb0.5Sn0.5I2Br interface, reducing the electron extraction potential barrier. Consequently, the planar solar cell with an ITO/PEDOT:PSS/CsPb0.5Sn0.5I2Br/PCBM/BCP/Ag layered structure featuring an F-TBA surface post-treatment demonstrated a noteworthy power conversion efficiency of 14.01%. Simultaneously, after being stored for 1000 h in an inert atmosphere glovebox, the non-encapsulated CsPb0.5Sn0.5I2Br solar cells managed to preserve 94% of their original efficiency.


Characterization of ACE and ACE2 Expression within Different Organs of the NOD Mouse.
Renin angiotensin system (RAS) is known to play a key role in several diseases such as diabetes, and renal and cardiovascular pathologies. Its blockade has been demonstrated to delay chronic kidney disease progression and cardiovascular damage in diabetic patients. In this sense, since local RAS has been described, the aim of this study is to characterize angiotensin converting enzyme (ACE) and ACE2 activities, as well as protein expression, in several tissues of the non-obese diabetic (NOD) mice model. After 21 or 40 days of diabetes onset, mouse serums and tissues were analyzed for ACE and ACE2 enzyme activities and protein expression. ACE and ACE2 enzyme activities were detected in different tissues. Their expressions vary depending on the studied tissue. Thus, whereas ACE activity was highly expressed in lungs, ACE2 activity was highly expressed in pancreas among the studied tissues. Interestingly, we also observed that diabetes up-regulates ACE mainly in serum, lung, heart, and liver, and ACE2 mainly in serum, liver, and pancreas. In conclusion, we found a marked serum and pulmonary alteration in ACE activity of diabetic mice, suggesting a common regulation. The increase of ACE2 activity within the circulation in diabetic mice may be ascribed to a compensatory mechanism of RAS.


MSX1 gene in the etiology orofacial deformities.
The muscle segment homeobox (MSX1) gene plays a crucial role in epithelial-mesenchymal tissue interactions in craniofacial development. It plays a regulative role in cellular proliferation, differentiation and cell death. The human MSX1 domain was also found in cow (Bt 302906), mouse (Mm 123311), rat (Rn13592001), chicken (Gg 170873) and clawed toad (XI 547690). Cleft lip and palate is the most common anomaly of the facial part of the skull. The etiology is not fully understood, but it is believed that the key role is played by the genetic factor activated by environmental factors. Among the candidate genes whose mutations could lead to formation of the cleft, the MSX1 homeobox gene is mentioned. Mutations in the gene MSX1 can lead to isolated cleft deformities, but also cause other dismorphic changes. Among the most frequently mentioned is loss of permanent tooth buds (mostly of less than 4 teeth - hypodontia, including second premolars). Mutations of MSX1 are observed in the Pierre- Robin sequence, which may be one of the features of congenital defects or is observed as an isolated defect. Mutation of the gene can lead to the occurrence of a rare congenital defect Wiktop (dental-nail) syndrome. Deletion of a fragment MSX1 (4p16.3) located in the WHS critical region, may be a cause of some symptoms of Wolf-Hirschhorn syndrome.


Hydrogen peroxide alters sternohyoid muscle function.
Upper airway (UA) dilator muscles are critical for the maintenance of airway patency. Injury or fatigue to this group of muscles, as observed in patients with obstructive sleep apnoea (OSA) and animal models of OSA, may leave the UA susceptible to collapse. Although the mechanisms underlying respiratory muscle dysfunction are not completely understood, there is strong evidence suggesting a link between increased production of reactive oxygen species and altered muscle function. The aim of this study was to examine the effects of H2O2 on rat sternohyoid muscle function in vitro. Sternohyoid contractile and endurance properties were examined at 35 °C under control or hypoxic conditions. Studies were conducted in the presence of varying concentrations of H2O2 (0, 0.01, 0.1 and 1 mM). Muscle function was also examined in the presence of antioxidants [desferoxamine (DFX), catalase] and the reducing agent dithiothreitol (DTT). H2O2 decreased muscle endurance in a concentration-dependent manner. This was partially reversed by catalase, DFX and DTT. Our results suggest that oxidants may contribute to UA respiratory muscle dysfunction with implications for the control of UA patency in vivo.


Prevalence and correlates of pre-diabetes in Sub-Saharan Africa using Demographic and Health Survey Data: a cross-sectional study.
The objective is to investigate the prevalence of pre-diabetes in Namibia and South Africa and to determine sociodemographic correlates of disease using population data. Cross-sectional study. Demographic and Health Survey for emerging (Namibia) and established (South Africa) economies in Sub-Saharan Africa collected laboratory data that allowed determination of pre-diabetes status. 3141 adults over age 18 from the 2013 Namibia survey, weighted to a population of 2176, and 4964 adults over age 18 from the 2016 South Africa survey, weighted to a population of 4627 had blood glucose/glycated haemoglobin (HbA1c) and diabetes information were included in the analysis. Pre-diabetes was defined as not being diagnosed with diabetes and having a blood sugar measurement of 100-125 mg/dL in Namibia or an HbA1c measurement of 5.7%-6.4%. Logistic models were run for each country separately, with pre-diabetes as the outcome and a series of sociodemographic variables (age, gender, urban/rural residence, number of children, employment status, wealth index, education level, and ethnicity (in South Africa) or religion (in Namibia)) entered as variables to investigate the independent relationship of each. The weighted prevalence of pre-diabetes was 18.7% in Namibia and 70.1% in South Africa. Rural residence was independently associated with higher odds of pre-diabetes in Namibia (1.47, 95% CI 1.05 to 2.06), while both younger age (0.98, 95% CI 0.97 to 0.99) and urban residence (0.80, 95% CI 0.66 to 0.99) were independently associated with odds of pre-diabetes in South Africa. The prevalence of pre-diabetes was 18.7% in Namibia and 70.1% in South Africa. Correlates of pre-diabetes differed between the two countries with rural residents having higher odds of pre-diabetes in Namibia and urban residents with higher odds in South Africa. Aggressive interventions, including population level education and awareness programmes, and individual level education and lifestyle interventions that account for country-specific contextual factors are urgently needed to prevent progression to diabetes.


Diagnostic ultrasound in the head and neck region.
Improvements in ultrasound (US) resolution have led to the application of this non-ionizing imaging modality in the diagnosis of soft tissue disorders affecting the head and neck region. Although operator experience is paramount in image interpretation, US has the advantage of being inexpensive, safe and widely available. Ultrasound is the first line imaging modality for suspected salivary gland tumours and is useful in evaluating superficial neck swellings, such as lymph node disease, soft tissue cysts and vascular malformations. Ultrasound also improves the success rate when used for guidance during fine-needle aspiration cytology (FNAC). This paper aims to provide the general practitioner with an overview of the basic principles of image formation, the advantages and disadvantages of US, and an outline of its indications for conditions that might present to the dental practitioner.


Nitric oxide synthase 2 genetic variation rs2297514 associates with a decreased susceptibility to extremity post-traumatic osteomyelitis in a Chinese Han population.
Previous studies have indicated that nitric oxide synthase 2 (NOS2) genetic variations are involved in delayed fracture healing and fracture non-union. Whether these genetic variants associate with the development of osteomyelitis (OM) remains unclear. Here, we analyzed the potential relationships between NOS2 genetic variations and the risk of developing post-traumatic OM (PTOM) in a Chinese Han population. Altogether 704 participants, including 336 PTOM patients and 368 healthy controls, were genotyped of rs2297514 and rs2248814 of the NOS2 gene using the SNaPshot genotyping method. Outcomes showed that the frequency of allele C of rs2297514 in the patient group was significantly lower than that in the control group (48.7% vs. 54.5%, P = 0.029, OR = 0.792, 95% CI 0.642 - 0.976). In addition, significant associations were found between rs2297514 and susceptibility to PTOM by the recessive model (P = 0.007, OR = 0.633, 95% CI 0.453 - 0.884), and the homozygous model (P = 0.039, OR = 0.648, 95% CI 0.429 - 0.979). Moreover, patients with the CC genotype of rs2297514 had lower inflammatory biomarkers levels than the TT genotype, especially for the C-reactive protein (CRP) level (median: 4.1 mg/L vs. 8.9 mg/L, P = 0.027). However, no significant relationship was noted between rs2248814 and the risk of developing PTOM. In this Chinese cohort, rs2297514 is correlated with a decreased risk of PTOM development, with genotype CC as a protective factor.


let-7 Contributes to Diabetic Retinopathy but Represses Pathological Ocular Angiogenesis.
The in vivo function of microRNAs (miRs) in diabetic retinopathy (DR) and age-related macular degeneration (AMD) remains unclear. We report here that let-7 family members are expressed in retinal and choroidal endothelial cells (ECs). In ECs, overexpression of let-7 by adenovirus represses EC proliferation, migration, and networking in vitro, whereas inhibition of the let-7 family with a locked nucleic acid (LNA)-anti-miR has the opposite effect. Mechanistically, silencing of the let-7 target HMGA2 gene mimics the phenotype of let-7 overexpression in ECs. let-7 transgenic (let-7-Tg) mice show features of nonproliferative DR, including tortuous retinal vessels and defective pericyte coverage. However, these mice develop significantly less choroidal neovascularization (CNV) compared to wild-type controls after laser injury. Consistently, silencing of let-7 in the eye increased laser-induced CNV in wild-type mice. Together, our data establish a causative role of let-7 in nonproliferative diabetic retinopathy and a repressive function of let-7 in pathological angiogenesis, suggesting distinct implications of let-7 in the pathogenesis of DR and AMD.


[Chemical composition of fetal hemoglobin in Sardinian newborns].
G gamma:A gamma composition of 54 newborns from Sardinia was determined by electrophoresis on polyacrylamide gels containing urea and Triton X-100. G gamma was present at a concentration of 67% with bimodal distribution (with mean values at 65 and 70% respectively). Concentration higher than 76% was not observed. When Hb F in peripheral blood was below 70%, G gamma:A gamma ratio correlates with Hb F%.


Semi-conductor detectors in output factor measurements.
Output factors are generally measured with cylindrical ionization chambers. It was investigated if Si-diodes of p-type instead could be used. The advantage would be the small detector size and the robust construction of the detector. Two types of diodes were studied, one with a shielding layer of tungsten specially made to reduce the excess response for scattered photons and one standard diode without any extra shielding. The measurements were performed at accelerating potentials between 4 and 50 MV and beam sizes between 4 cm x 4 cm and 40 cm x 40 cm. The results showed that both types of diodes are suitable for measurements of head scatter factors in mini-phantoms. However, the diodes were found inappropriate for measurement of output factors for large fields in extended water phantoms. For small fields (<10 cm x 10 cm) a small detector is advantageous and no errors due to the scatter contribution were seen. An cylindrical ionization chamber is the best choice for output factor measurements in extended water phantoms for large field sizes while diodes are an alternative in small fields. There were negligible differences between the detectors in head scatter measurements in mini phantoms.


Do Clinical Parameters Reflect Local Bone Metabolism in Heterotopic Ossification After Septic or Aseptic THA?
Heterotopic ossification (HO) is a common complication after THA. Although current research primarily focuses on treatment and prevention, little is known about the local bone metabolism of HO and clinical contributing factors. We aimed to assess bone remodeling processes in HO using histomorphometry, focusing on the effects of inflammation and prior NSAID treatment. Specifically, we asked: (1) Are HO specimens taken from patients with periprosthetic joint infection (PJI) more likely to exhibit active bone modeling and remodeling than specimens taken at the time of revision from patients without infection? (2) Do clinical or inflammatory serum and synovial parameters reflect the microstructure of and remodeling in both HO entities? (3) Is NSAID treatment before revision surgery associated with altered local bone mineralization or remodeling properties? Between June 2021 and May 2022, we screened 395 patients undergoing revision THA at two tertiary centers in Germany. Of those, we considered all patients with radiographic HO as potentially eligible. Based on that, 21% (83 of 395) were eligible; a further 43 were excluded because of an inability to remove the implant intraoperatively (16 patients), insufficient material (11), comorbidities with a major effect on bone metabolism (10), or bone-specific drugs (six), leaving 10% (40) for analysis in this retrospective, comparative study. HO specimens were collected during aseptic (25 patients: 18 male, seven female, mean age 70 ± 11 years, mean BMI 29 ± 4 kg/m 2 ) and septic (15 patients: 11 male, four female, mean age 69 ± 9 years, mean BMI 32 ± 9 kg/m 2 ) revision THA at a mean of 6 ± 7 years after primary implantation and a mean age of 70 ± 9 years at revision. Septic origin (PJI) was diagnosed based on the 2018 International Consensus Meeting criteria, through a preoperative assessment of serum and synovial parameters. To specify the local bone microstructure, ossification, and cellular bone turnover, we analyzed HO specimens using micro-CT and histomorphometry on undecalcified sections. Data were compared with those of controls, taken from femoral neck trabecular bone (10 patients: five female, five male, mean age 75 ± 6 years, mean BMI 28 ± 4 kg/m 2 ) and osteophytes (10 patients: five female, five male, mean age 70 ± 10 years, mean BMI 29 ± 7 kg/m 2 ). The time between primary implantation and revision (time in situ), HO severity based on the Brooker classification, and serum and synovial markers were correlated with HO microstructure and parameters of cellular bone turnover. In a subgroup of specimens of patients with NSAID treatment before revision, osteoid and bone turnover indices were evaluated and compared a matched cohort of specimens from patients without prior NSAID treatment. Patients with aseptic and septic HO presented with a higher bone volume (BV/TV; aseptic: 0.41 ± 0.15, mean difference 0.20 [95% CI 0.07 to 0.32]; septic: 0.43 ± 0.15, mean difference 0.22 [95% CI 0.08 to 0.36]; femoral neck: 0.21 ± 0.04; both p < 0.001), lower bone mineral density (aseptic: 809 ± 66 mg HA/cm 3 , mean difference -91 mg HA/cm 3 [95% CI -144 to -38]; septic: 789 ± 44 mg HA/cm 3 , mean difference -111 mg HA/cm 3 [95% CI -169 to -53]; femoral neck: 899 ± 20 mg HA/cm 3 ; both p < 0.001), and ongoing bone modeling with endochondral ossification and a higher proportion of woven, immature bone (aseptic: 25% ± 17%, mean difference 25% [95% CI 9% to 41%]; septic: 37% ± 23%, mean difference 36% [95% CI 19% to 54%]; femoral neck: 0.4% ± 0.5%; both p < 0.001) compared with femoral neck specimens. Moreover, bone surfaces were characterized by increased osteoblast and osteoclast indices in both aseptic and septic HO, although a higher density of osteocytes was detected exclusively in septic HO (aseptic: 158 ± 56 1/mm 2 versus septic: 272 ± 48 1/mm 2 , mean difference 114 1/mm 2 [95% CI 65 to 162]; p < 0.001). Compared with osteophytes, microstructure and turnover indices were largely similar in HO. The Brooker class was not associated with any local bone metabolism parameters. The time in situ was negatively associated with bone turnover in aseptic HO specimens (osteoblast surface per bone surface: r = -0.46; p = 0.01; osteoclast surface per bone surface: r = -0.56; p = 0.003). Serum or synovial inflammatory markers were not correlated with local bone turnover in septic HO. Specimens of patients with NSAID treatment before revision surgery had a higher osteoid thickness (10.1 ± 2.1 µm versus 5.5 ± 2.6 µm, mean difference -4.7 µm [95% CI -7.4 to -2.0]; p = 0.001), but there was no difference in other osteoid, structural, or cellular parameters. Aseptic and septic HO share phenotypic characteristics in terms of the sustained increase in bone metabolism, although differences in osteocyte and adipocyte numbers suggest distinct homeostatic mechanisms. These results suggest persistent bone modeling or remodeling, with osteoblast and osteoclast indices showing a moderate decline with the time in situ in aseptic HO. Future studies should use longitudinal study designs to correlate our findings with clinical outcomes (such as HO growth or recurrence). In addition, the molecular mechanisms of bone cell involvement during HO formation and growth should be further investigated, which may allow specific therapeutic and preventive interventions. To our knowledge, our study is the first to systematically investigate histomorphometric bone metabolism parameters in patients with HO after THA, providing a clinical reference for evaluating modeling and remodeling activity. Routine clinical, serum, and synovial markers are not useful for inferring local bone metabolism.


Tyrosine hydroxylase expression in differentiating neurons of the rat arcuate nucleus: stimulatory influence of serotonin afferents.
The influence of serotonin afferents on tyrosine hydroxylase expression in differentiating neurons of the rat arcuate nucleus was studied in vivo and in vitro. In the in vivo study, pchlorophenylalanine inhibited serotonin synthesis in fetal brain from the 11th to the 20th embryonic day. We then used semiquantitative immunocytochemistry to evaluate tyrosine hydroxylase levels in neurons of the arcuate nucleus in fetuses at the 21st embryonic day or in offspring at the 35th postnatal day. Serotonin depletion significantly decreased the tyrosine hydroxylase content in neurons of males and females at the 21st embryonic day and in males at the 35th postnatal day. For the in vitro study, embryonic neurons of the arcuate nucleus were cocultured with embryonic neurons of the raphe nucleus, the main source of serotonin innervation of the brain, including the arcuate nucleus. Co-culture of the neurons resulted in a gender-specific increase of the tyrosine hydroxylase level in the neurons of the arcuate nucleus. In turn, the neurons of the raphe nucleus showed increased levels of serotonin in both males and females, with no sexual dimorphism. Thus, our results suggest a stimulatory, long-lasting effect of serotonin afferents on tyrosine hydroxylase expression in the differentiating neurons of the rat arcuate nucleus during prenatal ontogenesis.


Photonic-plasmonic hybrid single-molecule nanosensor measures the effect of fluorescent labels on DNA-protein dynamics.
Current methods to study molecular interactions require labeling the subject molecules with fluorescent reporters. However, the effect of the fluorescent reporters on molecular dynamics has not been quantified because of a lack of alternative methods. We develop a hybrid photonic-plasmonic antenna-in-a-nanocavity single-molecule biosensor to study DNA-protein dynamics without using fluorescent labels. Our results indicate that the fluorescein and fluorescent protein labels decrease the interaction between a single DNA and a protein due to weakened electrostatic interaction. Although the study is performed on the DNA-XPA system, the conclusion has a general implication that the traditional fluorescent labeling methods might be misestimating the molecular interactions.


Role of adenosine in regulating the heterogeneity of skeletal muscle blood flow during exercise in humans.
Evidence from both animal and human studies suggests that adenosine plays a role in the regulation of exercise hyperemia in skeletal muscle. We tested whether adenosine also plays a role in the regulation of blood flow (BF) distribution and heterogeneity among and within quadriceps femoris (QF) muscles during exercise, measured using positron emission tomography. In six healthy young women, BF was measured at rest and then during three incremental low and moderate intermittent isometric one-legged knee-extension exercise intensities without and with theophylline-induced nonselective adenosine receptor blockade. BF heterogeneity within muscles was calculated from 16-mm(3) voxels in BF images and heterogeneity among the muscles from the mean values of the four QF compartments. Mean BF in the whole QF and its four parts increased, and heterogeneity decreased with workload both without and with theophylline (P < 0.001). Adenosine receptor blockade did not have any effect on mean bulk BF or BF heterogeneity among the QF muscles, yet blockade increased within-muscle BF heterogeneity in all four QF muscles (P = 0.03). Taken together, these results show that BF becomes less heterogeneous with increasing exercise intensity in the QF muscle group. Adenosine seems to play a role in muscle BF heterogeneity even in the absence of changes in bulk BF at low and moderate one-leg intermittent isometric exercise intensities.


Multiple endocrine neoplasias in a dog: corticotrophic tumour, bilateral adrenocortical tumours, and pheochromocytoma.
In a 10-year-old ovariohysterectomized standard Schnauzer, the finding of dexamethasone-resistant hypersecretion of cortisol, the results of computed tomography, and elevated plasma concentrations of ACTH suggested the presence of both adrenocortical tumour and pituitary-dependent hyperadrenocorticism. The dog made an uneventful recovery after bilateral adrenalectomy and remained in good health for 31/2 years with substitution for the induced hypoadrenocorticism. Then the enlarged pituitary caused neurological signs and eventually euthanasia was performed. The surgically excised right adrenal contained a well-circumscribed tumour of differentiated adrenocortical tissue and in the left adrenal there were two adrenocortical tumours and a pheochromocytoma. The unaffected parts of the adrenal cortices were well developed and without regressive transformation. At necropsy there were no metastatic lesions. The cells of the pituitary tumour were immunopositive for ACTH and had characteristics of malignancy. The present combination of corticotrophic tumour, adrenocortical tumours, and pheochromocytoma may be called 'multiple endocrine neoplasia' (MEN), but does not correspond to the inherited combinations of diseases known in humans as the MEN-1 and the MEN-2 syndromes. It is suggested that the co-existence of hyperadrenocorticism and pheochromocytoma may be related to the vascular supply of the adrenals. Some chromaffin cells of the adrenal medulla are directly exposed to cortical venous blood, and intra-adrenal cortisol is known to stimulate catecholamine synthesis and may promote adrenal medullary hyperplasia or neoplasia.


Invited comment: lipids and the development of immune dysfunction and infection.
Excessive W-6 PUFA metabolism due to high levels of dietary fat intake can encourage infection via prolonged inflammation, enhanced Gram negative survival, reticuloendothelial blockage, immunosuppression, and monokine depression. Lipids can influence host immunity by altering eicosanoid metabolism and membrane structure and function. Further investigations are essential to answer questions regarding the levels and properties of various essential fatty acids in TPN lipid emulsions. Combining the features of LCT in the form of W-3 PUFA (fish oil) and MCT in the form of medium-chain triglyceride in a "structured lipid" may decrease infection and may improve survival rates by producing fewer inflammatory eicosanoids of the two- and four-series, and serving as a more "efficient fuel." The introduction of W-3 polyunsaturated fatty acids into the TPN emulsions as well as into normal diets may provide an important therapeutic advance in the pathogenesis of disease. Such unique antiinflammatory properties of W-3 PUFA require intensive research.


The Impact of Just and Unjust War Events on Mental Health Need and Utilization within U.S. Service Members.
Background: Soldiers are resilient to just war events, such as killing enemy combatants and life-threatening experiences, but these same soldiers appear to struggle with unjust war events, such as killing a noncombatant or being unable to help civilian women and children in need. This study is the first to examine how just and unjust war experiences are associated with clinical health service outcomes. Methods: Two samples of soldiers in different stages of readjustment from deployment were drawn from a longitudinal, survey-based study of a US Army brigade. Measures included items related to combat events, mental health utilization, perceived mental health need, PTSD, depression, and functional impairment. Results: After controlling for other kinds of combat events, just war events (i.e., life-threatening events and killing enemy combatants) predicted outcomes in soldiers who are less than three months post-deployment, but only predicted 2 of 26 outcomes in soldiers one year post deployment. In contrast, unjust war events were found to be robust predictors of short-term and long-term outcomes related to mental health need and utilization, even after controlling for exposure to other combat events. Conclusions: The results extend previous longitudinal research that suggests that exposure to unjust war events carry a heavier long-term mental health burden than other types of events. Additionally, Soldiers exposed to unjust war events had an unmet need for care one year post deployment that was not directly tied to PTSD or depression. The results question the emphasis on life-threat within mental health pathogenesis models.


Loss of AP-2delta reduces retinal ganglion cell numbers and axonal projections to the superior colliculus.
AP-2δ is the most divergent member of the Activating Protein-2 (TFAP2) family of transcription factors. AP-2δ is restricted to specific regions of the CNS, including a subset of ganglion cells in the retina. Retinal ganglion cells (RGCs), the only output neurons of the retina, are responsible for transmitting the visual signal to the brain. AP-2δ knockout results in loss of Brn3c (Pou4f3) expression in AP-2δ -positive RGCs. While AP-2δ-/- mice have morphologically normal retinas at birth, there is a significant reduction in retinal ganglion cell numbers by P21, after eye opening. Chromatin immunoprecipitation indicates that Brn3c is a target of AP-2δ in the retina. Using fluorochrome-conjugated cholera toxin subunit B to trace ganglion cell axons from the eye to the major visual pathways in the brain, we found 87 % and 32 % decreases in ipsilateral and contralateral projections, respectively, to the superior colliculus in AP-2δ-/- mice. In agreement with anatomical data, visually evoked responses recorded from the brain confirmed that retinal outputs to the brain are compromised. AP-2δ is important for the maintenance of ganglion cell numbers in the retina. Loss of AP-2δ alters retinal axonal projections to visual centers of the brain, with ipsilaterial projections to the superior colliculus being the most dramatically affected. Our results have important implications for integration of the visual signal at the superior colliculus.


A close temporal relationship of liver disease to antiribosomal P0 protein antibodies and central nervous system disease in patients with systemic lupus erythematosus.
To determine whether there is a close temporal relationship of liver disease to serum IgG and/or IgM antiribosomal P0 protein antibodies (anti-P0) and central nervous system (CNS) lupus in patients with systemic lupus erythematosus (SLE). The study included 70 patients with active SLE. Of these, 30 had IgG and/or IgM anti-P0 and 14 had CNS lupus other than psychiatric disease (nonpsychiatric CNS lupus). Of these 14 patients, 11 had anti-P0. Laboratory manifestations of liver disease were retrospectively analyzed. Liver disease not attributed to any cause other than SLE (SLE liver disease) was present in 8 of the 11 patients with anti-P0 with nonpsychiatric CNS lupus (72.7%), in none of the 19 patients with anti-P0 without nonpsychiatric CNS lupus (0%), and in one of the 40 patients without anti-P0 (2.5%). The prevalence of SLE liver disease was significantly greater in patients with anti-P0 with nonpsychiatric CNS lupus than in the other 2 groups (p < 0.0001). Mean levels of liver enzymes (lactate dehydrogenase, glutamic oxaloacetic transaminase, glutamate pyruvate transaminase, gammaglutamyl transpeptidase) were significantly higher in patients with anti-P0 with nonpsychiatric CNS lupus than in the other 2 groups. Serial studies in 3 patients showed that the appearance of anti-P0 and liver dysfunction slightly preceded the onset of nonpsychiatric CNS lupus. Anti-P0 may be related to the pathogenesis of CNS lupus and SLE liver disease found simultaneously in SLE. The appearance of anti-P0 and liver dysfunction may predict onset of CNS lupus.


Combinatorial effects of quercetin and sex-steroids on fluid and electrolytes' (Na+, Cl-, HCO3-) secretory mechanisms in the uterus of ovariectomised female Sprague-Dawley rats.
Dysregulation of uterine fluid environment could impair successful reproduction and this could be due to the effect of environmental estrogens. Therefore, in this study, effect of quercetin, an environmental estrogen on uterine fluid and electrolytes concentrations were investigated under sex-steroid influence. Ovariectomised adult female Sprague-Dawley rats were given 10, 50 or 100mg/kg/day quercetin subcutaneously with 17-β estradiol (E) for seven days or three days E, then three days E plus progesterone (P) (E+P) treatment. Uterine fluid secretion rate, Na+, Cl- and HCO3- concentrations were determined by in-vivo perfusion. Following sacrifice, uteri were harvested and levels of the proteins of interest were identified by Western blotting and Realtime PCR. Distribution of these proteins in the uterus was observed by immunofluorescence. Levels of uterine cAMP were measured by enzyme-linked immunoassay (EIA). Administration of quercetin at increasing doses increased uterine fluid secretion rate, Na+, Cl- and HCO3- concentrations, but to the levels lesser than that of E. In concordant, levels of CFTR, SLC4A4, ENaC (α, β and γ), Na+/K+-ATPase, GPα/β, AC and cAMP in the uterus increased following increased in the doses of quercetin. Co-administration of quercetin with E caused uterine fluid secretion rate, Na+, Cl- and HCO3- concentrations to decrease. In concordant, uterine CFTR, SLC26A6, SLC4A4, ENaC (α, β and γ), Na+/K+-ATPase, GPα/β, AC and cAMP decreased. Greatest effects were observed following co-administration of 10mg/kg/day quercetin with E. Co-administration of quercetin with E+P caused uterine fluid Na+ and HCO3- concentrations to increase but no changes in fluid secretion rate and Cl- concentration were observed. Co-administration of high dose quercetin (100 mg/kg/day) with E+P caused uterine CFTR, SLC26A6, AC, GPα/β and ENaC (α, β and γ) to increase. Quercetin-induced changes in the uterine fluid secretion rate and electrolytes concentrations could potentially affect the uterine reproductive functions under female sex-steroid influence.


Eosinophilic Angiocentric Fibrosis as a Stenosing Lesion in the Subglottis.
Subglottic Eosinophilic Angiocentric Fibrosis (EAF) is an extremely rare disease of an elusive aetiology. It is chronically progressive benign condition that causes narrowing of the subglottic region leading to dysphonia and airway compromise. The diagnosis is historical and imaging is nonspecific. We report a case xc of 56-year-old lady referred to our institution with globus sensation, hoarseness, and mild stridor. Incidental subglottic mass was found at time of diagnostic microlaryngoscopy and biopsy confirmed subglottic EAF. All laboratory investigations were unremarkable. Lesion was removed with laryngeal microdebrider and three courses of intravenous dexamethasone were administered. Patient's postoperative period was uneventful and had remained disease free for 1 year. To date, no consensus has been reached on the optimal treatment of subglottic EAF. We recommend regular follow-up to detect early recurrence.


Primary ovarian carcinoid tumors may express CDX-2: a potential pitfall in distinction from metastatic intestinal carcinoid tumors involving the ovary.
Carcinoid tumors of the ovary are rare neoplasms that may be primary or metastatic. Clinicopathologic features such as unilaterality and early stage favor a primary ovarian neoplasm but in the absence of other teratomatous elements it may be difficult or impossible to determine whether an ovarian carcinoid is primary or metastatic. CDX-2 is a marker of intestinal differentiation that has been proposed as a marker of midgut origin for metastatic carcinoids. Its expression has not been tested in ovarian carcinoids. Additional markers of potential help in defining the origin of a carcinoid include cytokeratin (CK) 20, CK7, and thyroid transcription factor (TTF-1), none of which have been studied in ovarian carcinoids. We evaluated the diagnostic utility of CDX-2, CK20, CK7, and TTF-1 as well as conventional clinicopathologic features in determining the site of origin in 26 ovarian carcinoids (16 primary and 10 metastatic from midgut). Non-neoplastic premenopausal ovaries (n=10) served as controls. All primary ovarian carcinoids were unilateral whereas only 3/10 metastatic carcinoids were unilateral. Multinodular growth occurred in 6/10 metastatic carcinoids but not in any primary carcinoid. The average size of primary ovarian carcinoids was 3.4 cm (range: 0.2-13.5 cm) versus 10.2 cm for metastatic carcinoids (range: 4-32 cm). Of the primary ovarian carcinoids, 12/16 were 3 cm or smaller whereas all metastatic carcinoids were 4 cm or larger. Teratomatous elements were present in association with 10/16 primary ovarian carcinoids, whereas none were present in any metastatic carcinoid. The primary ovarian carcinoid types were insular (n=6), trabecular (n=3), strumal (n=6, of which 5 were trabecular pattern and 1 was insular pattern) or mucinous (n=1). CDX-2 was not expressed in any cells in normal ovaries. Among primary ovarian neoplasms, there was diffuse nuclear CDX-2 expression in 4/6 insular, 0/3 trabecular, 1/6 strumal (1/1 insular pattern and 0/5 trabecular pattern strumal carcinoids), and 1/1 mucinous carcinoids. All metastatic carcinoids, except for two of mucinous type, were insular. CDX-2 was diffusely and strongly expressed in all 8 metastatic insular carcinoids and in both metastatic mucinous carcinoids. None of the metastases was trabecular in type but 12 primary hindgut or foregut trabecular carcinoids were evaluated and all were negative for CDX-2. None of the ovarian carcinoids expressed TTF-1, CK7, or CK20, except for the primary and metastatic mucinous carcinoids, all of which were CK20-positive. These results demonstrate that CDX-2 cannot be used to determine if a carcinoid is primary in the ovary or metastatic from the intestine as insular and mucinous types of either origin express this marker. Trabecular carcinoids of either origin lack CDX-2 expression. CK20, CK7, or TTF-1 do not have diagnostic utility in this context. Conventional clinicopathologic features (unilaterality, lack of multinodular growth, early stage, presence of teratomatous elements, and size 3 cm or smaller) are the most helpful findings in suggesting a primary origin for an ovarian carcinoid tumor.


Chinese villages and their sustainable future: the European Union-China-Research Project "SUCCESS".
China has 800,000 villages-one person out of seven on the globe is living in a Chinese rural settlement. Yet the global discussions about the situation in China is currently characterised by a disproportionate focus on the development of towns and until now circumstances have generally been neglected in the rural areas, where 70% of the Chinese population is still living. Within the 5 years of the SUCCESS project research, this set of actual problems has been considered and analysed under the principle of sustainability: "What to maintain?" "What to change?" were the overall research questions asked in the SUCCESS project; the researchers were looking for answers under a sustainability regime, respecting the need to raise the quality of life in the villages. Several interweaving processes were used to achieve results: the inter-disciplinary research process between many areas of expertise, the trans-disciplinary process between the researchers and the Chinese villagers, and a negotiation process that made the connection between these two processes. The introduction describes the basic sustainability definition that was orienting the whole study. The innovation lays mostly in the methodology: the inter-disciplinary research co-operation related to practice and to involving the affected communities is needed to manage the significant and growing imbalances between urban and rural areas regarding their sustainability. In the transdisciplinary work, the project developed "village future sentences" that describe the local outcome of the research as one step towards better theoretical understanding of the mechanisms that could lead to a sustainable future, and they also managed to start sustainability processes in the case study sites. The integrated approach of the project helped generating future scenarios for these villages covering all aspects of their development, including urban design issues. Out of these scenarios, the villages developed small projects that could be implemented during the research period. This work made an important impact on community thinking within these villages. However, it can also be seen as contributing to the dramatically changing development process in China, by finding a balance between traditional and contemporary approaches. In particular, the approach demonstrated a new, inter-disciplinary and trans-disciplinary negotiation processes whereby the local knowledge and the expert knowledge find common ground and outcomes. The article follows the hypothesis that only comprehensive concepts can contribute to an upgraded living standard, where living spaces and rural life should be recognised and esteemed in the future as a complement to urban lifestyles within the Chinese society. Innovative knowledge generation-such as the "systemic structure constellation" technique or the systems model approach-helped to bring out latent needs, hopes and potential of the villagers. Besides the practical usage of these implemented projects, the process leading there showed the stakeholders their own fields of action. One major impact of these projects is the visibility of the results, which is crucial for villagers' awareness, their self-confidence and their experience with a successful participation in decision-making processes. Another impact is the potential for replicating results of sustainability-oriented patterns throughout China, especially as three of the villages have been nominated official model villages. Scenarios of a sustainable future for Chinese villages were the objective of the SUCCESS project. The first condition for this future is the question whether they can persist into the future-and to picture the importance of the rural environment and living space as a relevant element of Chinese life that needs to get a better image and more attention from the authorities and from the public opinion. Therefore, the final sentence that the whole research consortium, composed of 17 scientific institutions from European Union and China, agreed upon as a common result for the SUCCESS project, is as follows: "China is composed of a rich diversity of villages with many attractive qualities and essential resources for the future growth of the whole country; we recommend that policy makers cherish the human and natural potential of the rural economy and environment so that villages provide the foundation for sustainable development of this progressive nation" [Dumreicher, H., 2006. SUCCESS-a sustainable future for Chinese villages. International Symposium "Chinese Villages and their Sustainable Future", University of Natural Resources and Applied Life Sciences, Vienna, January 16]. This sentence was used in papers that where sent to different Chinese authorities by the Chinese partners and found its way, as a sort of "unofficial Charta", towards governmental agencies at national and provincial levels. The team carried out a 5-year-research study in rural China, aiming at establishing future images under the premises of sustainability. But the basic topic that needed to be tackled with was the question whether at all those villages could persist in the coming decades of rapid development. Therefore, the first aim of the study was to establish the importance of the rural environment and living space as a basis for the future of China.


Successful resuscitation after carbon dioxide embolism during laparoscopy.
Venous air embolism (VAE) is a rare life-threatening complication that can occur during laparoscopy. A 50-year-old previously healthy woman underwent laparoscopic cholecystectomy and liver cyst fenestration. Immediately after the surgeon had left the operating room, the patient became hypotensive and developed cardiac arrest. Resuscitation was initiated and a precordial ultrasound examination suspected VAE in the right cardiac chambers. The patient was positioned in Durant's position and air was aspirated through a central venous line. The patient was resuscitated and stabilized, and was transferred to another hospital, where she received hyperbaric oxygen treatment. The patient was discharged 14 days after surgery without any sequelae. It is important that the general surgeon suspects VAE during laparoscopy whenever the patient develops sudden and unexplained severe hypotension or cardiac arrest during or immediately after laparoscopy.


Impaired hemorrhage tolerance in the obese Zucker rat model of metabolic syndrome.
As obese Zucker rats (OZR) manifesting the metabolic syndrome exhibit enhanced vascular adrenergic constriction and potentially an enhanced adrenergic activity vs. lean Zucker rats (LZR), this study tested the hypothesis that OZR exhibit an improved tolerance to progressive hemorrhage. Preliminary experiments indicated that, corrected for body mass, total blood volume was reduced in OZR vs. LZR. Anesthetized LZR and OZR had a cremaster muscle prepared for in situ videomicroscopy and had renal, splanchnic, hindlimb, and skeletal muscle perfusion monitored with flow probes. Arterial pressure, arteriolar reactivity to norepinephrine, and tissue/organ perfusion were monitored after either infusion of phentolamine or successive withdrawals of 10% total blood volume. Phentolamine infusion indicated that regional adrenergic tone under control conditions differs substantially between LZR and OZR, whereas with hemorrhage OZR exhibit decompensation in arterial pressure before LZR. Renal, distal hindlimb, and skeletal muscle perfusion decreased more rapidly and to a greater extent in OZR vs. LZR after hemorrhage. In contrast, hemorrhage-induced reductions in splanchnic perfusion in OZR lagged behind those in LZR, although a similar maximum reduction was ultimately attained. With increasing hemorrhage, cremasteric arteriolar tone increased more in OZR than LZR, and this increase in active tone was entirely due to an elevated adrenergic contribution. Norepinephrine-induced arteriolar constriction was greater in OZR vs. LZR under control conditions and during hemorrhage, with arterioles from OZR demonstrating early closure vs. LZR. These results suggest that a combination of reduced blood volume and elevated peripheral adrenergic constriction contribute to impaired hemorrhage tolerance in OZR.


Detection of chikungunya virus antigen by a novel rapid immunochromatographic test.
Chikungunya fever is a mosquito-borne disease of key public health importance in tropical and subtropical countries. Although severe joint pain is the most distinguishing feature of chikungunya fever, diagnosis remains difficult because the symptoms of chikungunya fever are shared by many pathogens, including dengue fever. The present study aimed to develop a new immunochromatographic diagnosis test for the detection of chikungunya virus antigen in serum. Mice were immunized with isolates from patients with Thai chikungunya fever, East/Central/South African genotype, to produce mouse monoclonal antibodies against chikungunya virus. Using these monoclonal antibodies, a new diagnostic test was developed and evaluated for the detection of chikungunya virus. The newly developed diagnostic test reacted with not only the East/Central/South African genotype but also with the Asian and West African genotypes of chikungunya virus. Testing of sera from patients suspected to have chikungunya fever in Thailand (n = 50), Laos (n = 54), Indonesia (n = 2), and Senegal (n = 6) revealed sensitivity, specificity, and real-time PCR (RT-PCR) agreement values of 89.4%, 94.4%, and 91.1%, respectively. In our study using serial samples, a new diagnostic test showed high agreement with the RT-PCR within the first 5 days after onset. A rapid diagnostic test was developed using mouse monoclonal antibodies that react with chikungunya virus envelope proteins. The diagnostic accuracy of our test is clinically acceptable for chikungunya fever in the acute phase.


The Effect of Ultrasound-Assisted Debridement Combined with Vacuum Pump Therapy in Deep Sternal Wound Infections.
To assess whether a combined treatment of low-frequency (25 kHz) ultrasonic-debridement systems followed by vacuum-assisted wound closure (VAC) produces a better outcome in deep sternal wound infections (SWIs) compared to that of VAC alone. We evaluated 45 consecutive patients (25 males) between January 2013 and December 2016, in whom deep SWI was treated with a combination of low-frequency ultrasonic debridement system followed by vacuum-assisted closure (group A, n = 23) or with only vacuum-assisted closure therapy (group B, n = 22). Our final step in both groups was a secondary wound closure with a musculocutaneous flap. In both groups, a similar variety of bacteria were isolated. The time between eradication and secondary wound closure was significantly shorter in group A (7.3 ± 4.8 vs. 19.9 ± 17.2 days, p = 0.001). After a third debridement session, 95.7% of microbiological cultures were negative in group A versus 54.5% in B ( p = 0.001). Duration of antibiotic treatment ( p = 0.003) and hospitalization time ( p = 0.0001) were significantly shorter in group A. The use of low-frequency ultrasonic debridement system is an effective, less invasive technique to combat wound infection. In combination with vacuum-assisted closure therapy, we documented good mid-term results in our patients.


Vapour and Liquid-Phase Artemisia annua Essential Oil Activities against Several Clinical Strains of Candida.
Candida spp. are often the cause of infection in immune-compromised individuals. They are characterized by a strong resistance to antimicrobial drugs and disinfectants. The activity of Artemisia annua essential oil against Candida spp. was determined by vapour contact and microdilution assay. The oil was characterized by the presence of oxygenated monoterpenes (more than 75 % of the constituents), mainly represented by the irregular monoterpene artemisia ketone (ca. 22 %), and the widespread monoterpenes 1,8 cineole (ca. 19 %) and camphor (ca. 17 %). Other representative constituents were artemisia alcohol (5.9 %), α-pinene (5.7 %), and pinocarvone (3.0 %). Thujone, a typical toxic constituent of the Artemisia species, was not detected. The results are reported as minimum inhibitory concentration, minimum fungicidal concentration, and diameter of inhibition zone obtained by the vapour diffusion assay. We tested 10 clinical Candida strains, coming from both clinical samples and international collections. The results show that the antifungal activity of A. annua is influenced by the type of method adopted. The inhibitory action of the essential oil was, in fact, higher in the vapour than in the liquid phase. Our results show an average minimum inhibitory concentration in the liquid phase of 11.88 µL/mL, while in the vapour phase, the growth of all Candida strains tested at a concentration of 2.13 µL/cm(3) was inhibited. A strain of Candida glabrata was found to be less susceptible to the liquid medium than the vapour assay (50 µL/mL vs. 0.64 µL/cm(3), respectively). Candida albicans and Candida dubliniensis were the most susceptible to the vapour test, while Candida parapsilosis was the most resistant.


Incidence of syphilis, active component, U.S. Armed Forces, 1 January 2010 through 31 August 2015.
In 2014,the Centers for Disease Control and Prevention reported significant increases in cases of primary and secondary syphilis in the U.S.; among beneficiaries of the Military Health System, monthly surveillance reports tracking reportable medical events of syphilis have reflected similar increases. This analysis reports on incident cases and rates of syphilis among active component service members of the U.S. Armed Forces from 1 January 2010 through 31 August 2015. During the surveillance period, 2,976 cases of syphilis were diagnosed. Crude incidence rates increased from 30.9 cases per 100,000 person-years (p-yrs) in 2010 to 47.4 cases per 100,000 p-yrs in 2015. Males accounted for 88.7% of cases. Incidence rates of syphilis were highest among service members who were black, non-Hispanic or who were aged 20-29 years. About one-quarter of syphilis cases (24.4%; 727 cases) were diagnosed as HIV infected. Primary and secondary syphilis cases comprised 42% of all syphilis cases. Increasing rates of primary and secondary syphilis in active component service members reflect similar trends reported in the U.S. civilian population.


Plant Promoters and Terminators for High-Precision Bioengineering.
High-precision bioengineering and synthetic biology require fine-tuning gene expression at both transcriptional and posttranscriptional levels. Gene transcription is tightly regulated by promoters and terminators. Promoters determine the timing, tissues and cells, and levels of the expression of genes. Terminators mediate transcription termination of genes and affect mRNA levels posttranscriptionally, e.g., the 3'-end processing, stability, translation efficiency, and nuclear to cytoplasmic export of mRNAs. The promoter and terminator combination affects gene expression. In the present article, we review the function and features of plant core promoters, proximal and distal promoters, and terminators, and their effects on and benchmarking strategies for regulating gene expression.


Detection of Neisseria gonorrhoeae and Chlamydia trachomatis in genitourinary specimens from men and women by a coamplification PCR assay.
A coamplification PCR test for the direct detection of Neisseria gonorrhoeae and Chlamydia trachomatis in urethral and endocervical swabs and urine samples from men and women was compared to standard culture techniques. Processed specimens were amplified in single reaction tubes containing primers for both organisms, and PCR products were detected by a colorimetric microwell plate hybridization assay specific for each pathogen. Of 344 specimens from men, 45 (13.1%) urine specimens were PCR positive for C. trachomatis, 51 (14.8%) urethral swab specimens were PCR positive, and 29 urethral swab specimens (8.4%) were culture positive. After analysis of discrepancies, the resolved sensitivity and specificity of PCR for C. trachomatis were 96.2 and 99.3%, respectively, in urethral swab specimens, compared to 88.2 and 98.6% for urine specimens. Of the 192 specimens from women, 28 (14.6%) urine specimens were PCR positive for C. trachomatis, 32 (16.7%) endocervical specimens were PCR positive, and 19 (9.9%) endocervical specimens were culture positive. After analysis of discrepancies, the resolved sensitivity and specificity of PCR for C. trachomatis for endocervical specimens were both 100% compared to 100 and 99.4%, respectively, for urine specimens from women. In men, 68 (19.8%) urine specimens were PCR positive for N. gonorrhoeae, 73 (21.2%) urethral swabs were PCR positive, and 59 (17.2%) urethral swabs were culture positive. After analysis of discrepancies, the resolved sensitivity and specificity of PCR for N. gonorrhoeae were 97.3 and 97.0%, respectively, for urethral specimens compared to 94.4 and 98.5% for urine specimens. In women, 18 (9.4%) urine specimens were PCR positive for N. gonorrhoeae, 23 (12.0%) were endocervical swab PCR positive, and 15 (7.8%) endocervical specimens were culture positive. After analysis of discrepancies, the resolved sensitivity and specificity of PCR for N. gonorrhoeae were 100 and 99.4%, respectively, for endocervical specimens compared to 90.0 and 95.9% for female urine specimens. These results indicate that a multiplex PCR is highly sensitive for detecting both C. trachomatis and N. gonorrhoeae from a single urine or genital swab, providing a more cost-effective way of screening multiple pathogens.


Low SP1 expression differentially affects intestinal-type compared with diffuse-type gastric adenocarcinoma.
Specificity protein 1 (SP1) is an essential transcription factor that regulates multiple cancer-related genes. Because aberrant expression of SP1 is related to cancer development and progression, we focused on SP1 expression in gastric carcinoma and its correlation with disease outcomes. Although patient survival decreased as SP1 expression increased (P<0.05) in diffuse-type gastric cancer, the lack of SP1 expression in intestinal-type gastric cancer was significantly correlated with poor survival (P<0.05). The knockdown of SP1 in a high SP1-expressing intestinal-type gastric cell line, MKN28, increased migration and invasion but decreased proliferation. Microarray data in SP1 siRNA-transfected MKN28 revealed that the genes inhibiting migration were downregulated, whereas the genes negatively facilitating proliferation were increased. However, both migration and invasion were decreased by forced SP1 expression in a low SP1-expressing intestinal-type gastric cell line, AGS. Unlike the intestinal-type, in a high SP1-expressing diffuse-type gastric cell line, SNU484, migration and invasion were decreased by SP1 siRNA. In contrast to previous studies that did not identify differences between the 2 histological types, our results reveal that low expression of SP1 is involved in cancer progression and metastasis and differentially affects intestinal-type compared with diffuse-type gastric adenocarcinoma.


Systemic assessment of arterial wall structure and function in metabolic syndrome.
Metabolic syndrome (MetS) is thought to increase cardiovascular risk. The aim of this study was to investigate whether the MetS and its components are associated with parameters of arterial stiffness, flow mediated dilatation (FMD) of the brachial artery, carotid intima-media thickness (IMT) and the presence of carotid plaques (CP) in middle-aged subjects. One-hundred eighty-six asymptomatic volunteers (40-65 years-old, 86 males and 100 females) without clinically overt cardiovascular disease were examined. MetS was defined according to the International Diabetes Federation consensus. The prevalence of MetS was 32.8% (61 subjects). Augmentation index (Alx) and carotid-radial pulse wave velocity (PWV) as parameters of arterial stiffness were assessed by applanation tonometry. FMD (as endothelial function) was determined using high resolution B-mode ultrasonography, as were IMT and CP. PWV was significantly elevated in the MetS group (9.20 +/- 1.08 vs. 8.70 +/- 1.02, p = 0.003). FMD was significantly lower in the MetS group (5.32 vs. 6.45%, p = 0.018). There was no statistically significant difference in AIx between subjects with and without MetS (23.97 +/- 8.08 vs. 23.30 +/- 9.75, p = 0.248, respectively). Multiple regression analysis has shown that the presence of MetS was a significant (p = 0.005) determinant of PWV but not FMD. IMT* was higher in the MetS group (0.08 [0.07-0.1] vs. 0.07 [0.06-0.08] p < 0.001). The presence of arterial hypertension was related to AIx (R2 = 0.444, adjusted R2 = 0.434, p = 0.001) and PWV (R2 = 0.161, adjusted R2 = 0.152, coefficient p = 0.001). Furthermore, mean arterial pressure (MAP) itself was an independent predictor of AIx (R2 = 0.571, adjusted R2 = 0.562, p < 0.001) and PWV (R2 = 0.281, adjusted R2 = 0.269, p < 0.001). Glycemia had an impact on IMT (R2 = 0.027, adjusted R2 = 0.022, p = 0.026). The MetS was a significant predictor of the presence of carotid plaques (OR = 0.341, 95 % CI [0.173 - 0.673], p = 0.002). Blood pressure is the most important determinant of structure and function of arteries.