The 'sweet' spot of cellular pluripotency: protein glycosylation in human pluripotent stem cells and its applications in regenerative medicine.
Human pluripotent stem cells (hPSCs) promise for the future of regenerative medicine. The structural and biochemical diversity associated with glycans makes them a unique type of macromolecule modification that is involved in the regulation of a vast array of biochemical events and cellular activities including pluripotency in hPSCs. The primary focus of this review article is to highlight recent advances in stem cell research from a glycobiological perspective. We also discuss how our understanding of glycans and glycosylation may help overcome barriers hindering the clinical application of hPSC-derived cells. A literature survey using NCBI-PubMed and Google Scholar was performed in 2014. Regenerative medicine hopes to provide novel strategies to combat human disease and tissue injury that currently lack effective therapies. Although progress in this field is accelerating, many critical issues remain to be addressed in order for cell-based therapy to become a practical and safe treatment option. Emerging evidence suggests that protein glycosylation may significantly influence the regulation of cellular pluripotency, and that the exploitation of protein glycosylation in hPSCs and their differentiated derivatives may lead to transformative and translational discoveries for regenerative medicine. In addition, hPSCs represent a novel research platform for investigating glycosylation-related disease.


Genome Analysis of the Fruiting Body-Forming Myxobacterium Chondromyces crocatus Reveals High Potential for Natural Product Biosynthesis.
Here, we report the complete genome sequence of the type strain of the myxobacterial genus Chondromyces, Chondromyces crocatus Cm c5. It presents one of the largest prokaryotic genomes featuring a single circular chromosome and no plasmids. Analysis revealed an enlarged set of tRNA genes, along with reduced pressure on preferred codon usage compared to that of other bacterial genomes. The large coding capacity and the plethora of encoded secondary metabolite biosynthetic gene clusters are in line with the capability of Cm c5 to produce an arsenal of antibacterial, antifungal, and cytotoxic compounds. Known pathways of the ajudazol, chondramide, chondrochloren, crocacin, crocapeptin, and thuggacin compound families are complemented by many more natural compound biosynthetic gene clusters in the chromosome. Whole-genome comparison of the fruiting-body-forming type strain (Cm c5, DSM 14714) to an accustomed laboratory strain which has lost this ability (nonfruiting phenotype, Cm c5 fr-) revealed genetic changes in three loci. In addition to the low synteny found with the closest sequenced representative of the same family, Sorangium cellulosum, extensive genetic information duplication and broad application of eukaryotic-type signal transduction systems are hallmarks of this 11.3-Mbp prokaryotic genome.


Impaired renal concentrating ability during resuscitation from shock.
Renal concentrating ability was tested in 20 severely injured patients who received an average of 21 blood transfusions. Timed measurements were made of urine output (UO), and the clearance of creatinine glomerular filtration rate (GFR), sodium (CNa), osmols (COsm), and free water (CH2O) during operation and were repeated at 12 and 18 hours postoperatively, and on postoperative days 2 and 4. During operation the GFR was markedly reduced (36 mL/min), while UO, CNa, and COsm were all markedly increased (8.5, 5.9, and 8.1 mL/min, respectively). The CH2O was positive (0.4 mL/min). Following operation the rate of renal excretion of water and solutes was still high: UO, 4.0 mL/min; CNa, 4.3 mL/min; and COsm, 6.0 mL/min. Five hours postoperatively the CH2O had returned to normal. By day 2 the excretion rate of water had returned closer to normal: UO, 2.1 mL/min; CNa, 2.6 mL/min; COsm, 4.0 mL/min; the CH2O was normal. Subsequent study results were normal. These data demonstrate a renal concentrating impairment during and following operation. An osmotic diuresis, transient tubular ischemia, a washout of interstitial inner medullary osmoles, or some cryptic factor may be causative.


Up-regulation of MUC5AC and MUC5B mucin genes in chronic rhinosinusitis.
Excess mucus production and hypersecretion characterize upper airway diseases. The primary mechanisms leading to mucus hypersecretion in chronic rhinosinus inflammation are not well understood. Mucus hypersecretion is commonly accompanied by goblet cell and submucosal gland cell hyperplasia. It is important to identify which mucin gene messenger RNAs (mRNAs) are expressed in the sinus mucosa. To investigate the expression of MUC5AC and MUC5B mRNAs and localization of these proteins in human sinus mucosa and to compare the expression of MUC5AC and MUC5B mRNAs in normal and in chronic sinus mucosa. Twenty chronic maxillary sinusitis mucosa samples and 20 normal maxillary sinus mucosa samples were obtained; RNAs were extracted from sinus mucosa, and semiquantitative reverse transcription-polymerase chain reaction was performed for MUC5AC and MUC5B. Localization of these proteins was sought by using immunohistochemical analysis. The levels of MUC5AC and MUC5B mRNA in chronic rhinosinusitis were significantly increased compared with those in normal sinus mucosa (P =.02). In inflammed sinus mucosa, MUC5AC protein was expressed in the cytoplasm of the goblet cell in the surface epithelium, and MUC5B expression was restricted to mucous cells of the submucosal glands and to the epithelium of sinus mucosa. However, in the normal sinus mucosa, MUC5AC and MUC5B proteins were expressed at low levels in the sinus epithelium and submucosal glands, respectively. These results suggest that up-regulation of MUC5AC and MUC5B, which are major components of respiratory secretion in chronic rhinosinusitis, may play important roles in the pathogenesis of sinus hypersecretion in chronic rhinosinusitis.


Initiation of human lactation: secretory differentiation and secretory activation.
Theories for the origin of milk have been recorded since the time of Ancient Greeks. In those times it was believed that milk was derived from special vessels that connected the uterus to the breasts. The "chyle theory" on the origin of milk was another prominent theory which persisted well into the nineteenth century before the realisation that milk components were derived from blood and some milk constituents were actually synthesized within the breasts. The demonstration that milk ejection was the expulsion of milk that had already been secreted and that milk secretion was a separate continuous process, set the background for the development for the current understanding of milk synthesis and secretion. Today we know that there are two stages in the initiation of lactation- secretory differentiation and secretory activation. Secretory differentiation represents the stage of pregnancy when the mammary epithelial cells differentiate into lactocytes with the capacity to synthesize unique milk constituents such as lactose. This process requires the presence of a 'lactogenic hormone complex' of the reproductive hormones, estrogen, progesterone, prolactin and some metabolic hormones. Secretory activation on the other hand, is the initiation of copious milk secretion and is associated with major changes in the concentrations of many milk constituents. The withdrawal of progesterone triggers the onset of secretory activation but prolactin, insulin and cortisol must also be present. This review describes the works of pioneers that have led to our current understanding of the biochemical and endocrinological processes involved in the initiation of human lactation.


Parental metabolic syndrome epigenetically reprograms offspring hepatic lipid metabolism in mice.
The prevalence of nonalcoholic fatty liver disease (NAFLD) is increasing worldwide. Although gene-environment interactions have been implicated in the etiology of several disorders, the impact of paternal and/or maternal metabolic syndrome on the clinical phenotypes of offspring and the underlying genetic and epigenetic contributors of NAFLD have not been fully explored. To this end, we used the liver-specific insulin receptor knockout (LIRKO) mouse, a unique nondietary model manifesting 3 hallmarks that confer high risk for the development of NAFLD: hyperglycemia, insulin resistance, and dyslipidemia. We report that parental metabolic syndrome epigenetically reprograms members of the TGF-β family, including neuronal regeneration-related protein (NREP) and growth differentiation factor 15 (GDF15). NREP and GDF15 modulate the expression of several genes involved in the regulation of hepatic lipid metabolism. In particular, NREP downregulation increases the protein abundance of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) and ATP-citrate lyase (ACLY) in a TGF-β receptor/PI3K/protein kinase B-dependent manner, to regulate hepatic acetyl-CoA and cholesterol synthesis. Reduced hepatic expression of NREP in patients with NAFLD and substantial correlations between low serum NREP levels and the presence of steatosis and nonalcoholic steatohepatitis highlight the clinical translational relevance of our findings in the context of recent preclinical trials implicating ACLY in NAFLD progression.


A Randomized Controlled Trial of Clinician-Supported Problem-Solving Bibliotherapy for Family Caregivers of People With First-Episode Psychosis.
Family interventions for first-episode psychosis (FEP) are an integral component of treatment, with positive effects mainly on patients' mental state and relapse rate. However, comparatively little attention has been paid to the effects of family interventions on caregivers' stress coping and well-being, especially in non-Western countries. We aimed to test the effects of a 5-month clinician-supported problem-solving bibliotherapy (CSPSB) for Chinese family caregivers of people with FEP in improving family burden and carers' problem-solving and caregiving experience, and in reducing psychotic symptoms and duration of re-hospitalizations, compared with those only received usual outpatient family support (UOFS). A randomized controlled trial was conducted across 2 early psychosis clinics in Hong Kong, where there might be inadequate usual family support services for FEP patients. A total of 116 caregivers were randomly selected, and after baseline measurement, randomly assigned to the CSPSB or UOFS. They were also assessed at 1-week and 6- and 12-month post-intervention. Intention-to-treat analyses were applied and indicated that the CSPSB group reported significantly greater improvements in family burden and caregiving experience, and reductions in severity of psychotic symptoms and duration of re-hospitalizations, than the UOFS group at 6- and 12-month follow-up. CSPSB produces moderate long-term benefits to caregivers and FEP patients, and is a low-cost adjunct to UOFS.


Cost, staffing and quality impact of bedside electronic medical record (EMR) in nursing homes.
There is growing political pressure for nursing homes to implement the electronic medical record (EMR) but there is little evidence of its impact on resident care. The purpose of this study was to test the unique and combined contributions of EMR at the bedside and on-site clinical consultation by gerontological expert nurses on cost, staffing, and quality of care in nursing homes. Eighteen nursing facilities in 3 states participated in a 4-group 24-month comparison: Group 1 implemented bedside EMR, used nurse consultation; Group 2 implemented bedside EMR only; Group 3 used nurse consultation only; Group 4 neither. Intervention sites (Groups 1 and 2) received substantial, partial financial support from CMS to implement EMR. Costs and staffing were measured from Medicaid cost reports, and staff retention from primary data collection; resident outcomes were measured by MDS-based quality indicators and quality measures. Total costs increased in both intervention groups that implemented technology; staffing and staff retention remained constant. Improvement trends were detected in resident outcomes of ADLs, range of motion, and high-risk pressure sores for both intervention groups but not in comparison groups. Implementation of bedside EMR is not cost neutral. There were increased total costs for all intervention facilities. These costs were not a result of increased direct care staffing or increased staff turnover. Nursing home leaders and policy makers need to be aware of on-going hardware and software costs as well as costs of continual technical support for the EMR and constant staff orientation to use the system. EMR can contribute to the quality of nursing home care and can be enhanced by on-site consultation by nurses with graduate education in nursing and expertise in gerontology.


NSAIDs modulate clonal evolution in Barrett's esophagus.
Cancer is considered an outcome of decades-long clonal evolution fueled by acquisition of somatic genomic abnormalities (SGAs). Non-steroidal anti-inflammatory drugs (NSAIDs) have been shown to reduce cancer risk, including risk of progression from Barrett's esophagus (BE) to esophageal adenocarcinoma (EA). However, the cancer chemopreventive mechanisms of NSAIDs are not fully understood. We hypothesized that NSAIDs modulate clonal evolution by reducing SGA acquisition rate. We evaluated thirteen individuals with BE. Eleven had not used NSAIDs for 6.2±3.5 (mean±standard deviation) years and then began using NSAIDs for 5.6±2.7 years, whereas two had used NSAIDs for 3.3±1.4 years and then discontinued use for 7.9±0.7 years. 161 BE biopsies, collected at 5-8 time points over 6.4-19 years, were analyzed using 1Million-SNP arrays to detect SGAs. Even in the earliest biopsies there were many SGAs (284±246 in 10/13 and 1442±560 in 3/13 individuals) and in most individuals the number of SGAs changed little over time, with both increases and decreases in SGAs detected. The estimated SGA rate was 7.8 per genome per year (95% support interval [SI], 7.1-8.6) off-NSAIDs and 0.6 (95% SI 0.3-1.5) on-NSAIDs. Twelve individuals did not progress to EA. In ten we detected 279±86 SGAs affecting 53±30 Mb of the genome per biopsy per time point and in two we detected 1,463±375 SGAs affecting 180±100 Mb. In one individual who progressed to EA we detected a clone having 2,291±78 SGAs affecting 588±18 Mb of the genome at three time points in the last three of 11.4 years of follow-up. NSAIDs were associated with reduced rate of acquisition of SGAs in eleven of thirteen individuals. Barrett's cells maintained relative equilibrium level of SGAs over time with occasional punctuations by expansion of clones having massive amount of SGAs.


Herbicidal activity of cineole derivatives.
Essential oils and their constituents have potential as ecologically acceptable pesticides that may also have novel modes of action. In this work hydroxy and ester derivatives of the naturally occurring monoterpenoids 1,8-cineole 3, the main component in most eucalyptus oils, and 1,4-cineole 4 were prepared and their pre-emergence herbicidal activity against annual ryegrass (Lolium rigidum) and radish (Raphanus sativus var. Long Scarlet) investigated in laboratory-based bioassays. 1,8-Cineole, eucalyptus oil and all derivatives showed a dose-dependent herbicidal activity against annual ryegrass and radish with many of the derivatives showing improved herbicidal activity relative to 1,8-cineole and high-cineole eucalyptus oil. Increased activity of cineole ester derivatives compared to their associated hydroxy-cineole and carboxylic acid was not observed. No relationship between lipophilicity of the carboxylic acid portion of cineole ester derivatives and herbicidal activity was observed. The results indicate that these cineole derivatives could be environmentally acceptable herbicides.


Non-invasive evaluation of the haemodynamic effects of high-dose medetomidine in healthy cats for semen collection.
This study aimed to assess non-invasively the cardiovascular effects of high-dose medetomidine on healthy male cats undergoing semen collection. Haemodynamic evaluations were assessed on the basis of clinical examination, systolic arterial pressure (SAP) and transthoracic echocardiographic examination. Eight client owned, male domestic shorthair cats were sedated with a bolus of medetomidine intramuscularly (IM; 0.13 mg/kg), and semen collection was performed. A second transthoracic echocardiographic examination and SAP measurement were carried out 15 mins after sedation. At the end of the examination, the patients received a bolus of atipamezole (0.3 mg/kg) IM. The cats were deeply sedated, relaxed and laterally recumbent during the entire procedure. No rhythm abnormalities were observed during the examinations and no significant increase in SAP was recorded. Heart rate dropped from 200 ± 33 to 92 ± 13.1 beats per min after sedation. There was a significant increase in left ventricular dimensions and the left atrial area. The parameters of left ventricular systolic function were reduced, as were systemic and pulmonary cardiac outputs. Peak diastolic wave velocities were significantly reduced, while isovolumic contraction and relaxation time of the left ventricle were prolonged. Aortic valve insufficiency was recorded for all cats, while mitral valve insufficiency was noted in five cats. None of the subjects developed systolic anterior motion of the mitral valve. The protocol allowed us to collect good semen samples in healthy cats. However, high-dose medetomidine induces significant haemodynamic effects on the feline heart, mainly due to a reduced heart rate, an increased cardiac preload and impaired systolic function. The animals recovered from the anaesthesia, after antagonist administration, without showing any clinically relevant consequences.


Identification of two new mutations in the GPR98 and the PDE6B genes segregating in a Tunisian family.
Autosomal recessive retinitis pigmentosa (ARRP) is a genetically heterogeneous disorder. ARRP could be associated with extraocular manifestations that define specific syndromes such as Usher syndrome (USH) characterized by retinal degeneration and congenital hearing loss (HL). The USH type II (USH2) associates RP and mild-to-moderate HL with preserved vestibular function. At least three genes USH2A, the very large G-protein-coupled receptor, GPR98, and DFNB31 are responsible for USH2 syndrome. Here, we report on the segregation of non-syndromic ARRP and USH2 syndrome in a consanguineous Tunisian family, which was previously used to define USH2B locus. With regard to the co-occurrence of these two different pathologies, clinical and genetic reanalysis of the extended family showed (i) phenotypic heterogeneity within USH2 patients and (ii) excluded linkage to USH2B locus. Indeed, linkage analysis disclosed the cosegregation of the USH2 phenotype with the USH2C locus markers, D5S428 and D5S618, whereas the ARRP perfectly segregates with PDE6B flanking markers D4S3360 and D4S2930. Molecular analysis revealed two new missense mutations, p.Y6044C and p.W807R, occurring in GPR98 and PDE6B genes, respectively. In conclusion, our results show that the USH2B locus at chromosome 3p23-24.2 does not exist, and we therefore withdraw this locus designation. The combination of molecular findings for GPR98 and PDE6B genes enable us to explain the phenotypic heterogeneity and particularly the severe ocular affection first observed in one USH2 patient. This report presents an illustration of how consanguinity could increase familial clustering of multiple hereditary diseases within the same family.


Actin dynamics is controlled by a casein kinase II and phosphatase 2C interplay on Toxoplasma gondii Toxofilin.
Actin polymerization in Apicomplexa protozoa is central to parasite motility and host cell invasion. Toxofilin has been characterized as a protein that sequesters actin monomers and caps actin filaments in Toxoplasma gondii. Herein, we show that Toxofilin properties in vivo as in vitro depend on its phosphorylation. We identify a novel parasitic type 2C phosphatase that binds the Toxofilin/G-actin complex and a casein kinase II-like activity in the cytosol, both of which modulate the phosphorylation status of Toxofilin serine53. The interplay of these two molecules controls Toxofilin binding of G-actin as well as actin dynamics in vivo. Such functional interactions should play a major role in actin sequestration, a central feature of actin dynamics in Apicomplexa that underlies the spectacular speed and nature of parasite gliding motility.


An assessment of oral health importance: results of a statewide survey.
Barriers exist to the U.S. surgeon general's call to action toward a goal of achieving oral health for all Americans. The authors describe perceptions of oral health among the public according to the results of a statewide survey conducted in Minnesota. During a four-month period in 2005, 1,636 noninstitutionalized adults received a mixed-mode mail and telephone survey. The 40-item survey contained questions regarding the importance of oral health to respondents (overall), as well as the relative importance of oral health to overall health compared with diabetes, heart disease and arthritis. The majority of respondents (76.1 percent) rated the overall importance of their oral health as "very important." Interestingly, 58.2 percent, 45.8 percent and 69.2 percent of respondents believed that oral health was "about as important" as or "more important" than diabetes, heart disease and arthritis, respectively. Only sex and educational level were associated with self-reports of overall importance of oral health. The findings suggest that the majority of survey respondents considered oral health to be very important and equal in importance to certain medical conditions. Among survey respondents, men with less than a college education were less likely than others to value oral health. Clinicians should recognize that this may affect behavior in this patient group, especially among those who may not visit a dentist regularly. When researchers design interventions to address oral health perceptions among the public, health care professionals and/or policymakers, they need to take this patient group into account.


Transforming growth factor beta1 rescues serum deprivation-induced apoptosis via the mitogen-activated protein kinase (MAPK) pathway in macrophages.
Cell death and cell survival are central components of normal development and pathologic states. Transforming growth factor beta1 (TGF-beta1) is a pleiotropic cytokine that regulates both cell growth and cell death. To better understand the molecular mechanisms that control cell death or survival, we investigated the role of TGF-beta1 in the apoptotic process by dominant-negative inhibition of both TGF-beta1 and mitogen-activated protein kinase (MAPK) signaling pathways. Murine macrophages (RAW 264.7) undergo apoptosis following serum deprivation, as determined by DNA laddering assay. However, apoptosis is prevented in serum-deprived macrophages by the presence of exogenous TGF-beta1. Using stably transfected RAW 264.7 cells with the kinase-deleted dominant-negative mutant of TbetaR-II (TbetaR-IIM) cDNA, we demonstrate that this protective effect by TGF-beta1 is completely abrogated. To determine the downstream signaling pathways, we examined TGF-beta1 effects on the MAPK pathway. We show that TGF-beta1 induces the extracellular signal-regulated kinase (ERK) activity in a time-dependent manner up to 4 h after stimulation. Furthermore, TGF-beta1 does not rescue serum deprivation-induced apoptosis in RAW 264.7 cells transfected with a dominant-negative mutant MAPK (ERK2) cDNA or in wild type RAW 264.7 cells in the presence of the MAPK kinase (MEK1) inhibitor. Taken together, our data demonstrate for the first time that TGF-beta1 is an inhibitor of apoptosis in cultured macrophages and may serve as a cell survival factor via TbetaR-II-mediated signaling and downstream intracellular MAPK signaling pathway.


Culturing selects for specific genotypes of Borrelia burgdorferi in an enzootic cycle in Colorado.
In Colorado, Borrelia burgdorferi sensu stricto, the etiologic agent of Lyme disease, is maintained in an enzootic cycle between Ixodes spinipalpis ticks and Neotoma mexicana rats (27). The frequencies of flagellin (fla), 66-kDa protein (p66), and outer surface protein A (ospA) alleles were examined in 71 B. burgdorferi isolates from samples from Colorado. Approximately two-thirds of these samples were isolates from I. spinipalpis ticks that had been cultured in BSK-H medium prior to DNA extraction. The remaining samples were from total DNA extracted directly from infected I. spinipalpis ticks. A portion of each gene was amplified by PCR and screened for genetic variability by single-strand conformation polymorphism (SSCP) analysis. We identified three alleles in the fla gene, seven in the p66 gene, and seven in the ospA gene. Sequencing verified that the amplified products originated from B. burgdorferi template DNA and indicated 100% sensitivity and specificity of the SSCP analysis. The frequencies of the p66 and ospA alleles were significantly different between cultured and uncultured spirochetes. The number of three-locus genotypes and the genetic diversity of alleles at all loci were consistently lower in cultured spirochetes, suggesting that culturing of B. burgdorferi in BSK-H medium may select for specific genotypes.


Temporally controlled targeted somatic mutagenesis in embryonic surface ectoderm and fetal epidermal keratinocytes unveils two distinct developmental functions of BRG1 in limb morphogenesis and skin barrier formation.
Animal SWI2/SNF2 protein complexes containing either the brahma (BRM) or brahma-related gene 1 (BRG1) ATPase are involved in nucleosome remodelling and may control the accessibility of sequence-specific transcription factors to DNA. In vitro studies have indicated that BRM and BRG1 could regulate the expression of distinct sets of genes. However, as mice lacking BRM are viable and fertile, BRG1 might efficiently compensate for BRM loss. By contrast, as Brg1-null fibroblasts are viable but Brg1-null embryos die during the peri-implantation stage, BRG1 might exert cell-specific functions. To further investigate the in vivo role of BRG1, we selectively ablated Brg1 in keratinocytes of the forming mouse epidermis. We show that BRG1 is selectively required for epithelial-mesenchymal interactions in limb patterning, and during keratinocyte terminal differentiation, in which BRM can partially substitute for BRG1. By contrast, neither BRM nor BRG1 are essential for the proliferation and early differentiation of keratinocytes, which may require other ATP-dependent nucleosome-remodelling complexes. Finally, we demonstrate that cell-specific targeted somatic mutations can be created at various times during the development of mouse embryos cell-specifically expressing the tamoxifen-activatable Cre-ER(T2) recombinase.


Subwavelength confined terahertz waves on planar waveguides using metallic gratings.
A terahertz plasmonic waveguide is experimentally demonstrated using a plastic ribbon waveguide integrated with a diffraction metal grating to approach subwavelength-scaled confinement and long-distance delivery. Appropriately adjusting the metal-thickness and the periodical slit width of a grating greatly improves both guiding ability and field confinement in the hybrid waveguide structure. The measured lateral decay length of the bound terahertz surface waves on the hybrid waveguide can be reduced to less than λ/4 after propagating a waveguide of around 50mm-long in length. The subwavelength-confined field is potentially advantageous to biomolecular sensing or membrane detection because of the long interaction length between the THz field and analytes.


Evaluation of a flat panel digital radiographic system for low-dose portable imaging of neonates.
The purpose of this study was to evaluate the clinical utility of an investigational flat-panel digital radiography system for low-dose portable neonatal imaging. Thirty image-pairs from neonatal intensive care unit patients were acquired with a commercial Computed Radiography system (Agfa, ADC 70), and with the investigational system (Varian, Paxscan 2520) at one-quarter of the exposure. The images were evaluated for conspicuity and localization of the endings of ancillary catheters and tubes in two observer performance experiments with three pediatric radiologists and three neonatologists serving as observers. The results indicated no statistically significant difference in diagnostic quality between the images from the investigational system and from CR. Given the investigational system's superior resolution and noise characteristics, observer results suggest that the high detective quantum efficiency of flat-panel digital radiography systems can be utilized to decrease the radiation dose/exposure to neonatal patients, although post-processing of the images remains to be optimized. The rapid availability of flat-panel images in portable imaging was found to be an added advantage for timely clinical decision-making.


THE OXYGEN CONTENT OF THE BLOOD IN LOBAR PNEUMONIA.
In most cases of uncomplicated lobar pneumonia the decrease of respiratory surface is completely compensated for, and the oxygen content of the blood is within normal limits. Occasional cases of uncomplicated pneumonia have an oxygen content of the venous blood which is below normal. In the two cases reported here, this was associated with a carbon dioxide content of the blood which was higher than normally, and the condition was apparently due to an interference with the respiratory exchange of gases. In the terminal stage of the fatal cases of pneumonia in which death does not occur with great suddenness, there is often a progressive diminution in the oxygen content of the blood. Synchronous with this is a progressive decrease in the oxygen-combining capacity of the blood. These changes are usually seen in patients in whom an intense bacteremia has developed and are analogous to those found in the arterial blood of infected rabbits, and to those resulting from the growth of the pneumococcus in blood in vitro. In all three conditions there is probably a change of oxyhemoglobin to methemoglobin. This change of the hemoglobin molecule, so that it no longer takes up and gives off oxygen readily, is probably a factor in the immediate cause of death in many cases of pneumonia.


Recent advances in proteomic technologies applied to cardiovascular disease.
In recent years, the diagnosis of cardiovascular disease (CVD) has increased its potential, also thanks to mass spectrometry (MS) proteomics. Modern MS proteomics tools permit analyzing a variety of biological samples, ranging from single cells to tissues and body fluids, like plasma and urine. This approach enhances the search for informative biomarkers in biological samples from apparently healthy individuals or patients, thus allowing an earlier and more precise diagnosis and a deeper comprehension of pathogenesis, development and outcome of CVD to further reduce the enormous burden of this disease on public health. In fact, many differences in protein expression between CVD-affected and healthy subjects have been detected, but only a few of them have been useful to establish clinical biomarkers because they did not pass the verification and validation tests. For a concrete clinical support of MS proteomics to CVD, it is, therefore, necessary to: ameliorate the resolution, sensitivity, specificity, throughput, precision, and accuracy of MS platform components; standardize procedures for sample collection, preparation, and analysis; lower the costs of the analyses; reduce the time of biomarker verification and validation. At the same time, it will be fundamental, for the future perspectives of proteomics in clinical trials, to define the normal protein maps and the global patterns of normal protein levels, as well as those specific for the different expressions of CVD.


Left posterior BA37 is involved in object recognition: a TMS study.
Functional imaging studies have proposed a role for left BA37 in phonological retrieval, semantic processing, face processing and object recognition. The present study targeted the posterior aspect of BA37 to see whether a deficit, specific to one of the above types of processing could be induced. Four conditions were investigated: word and nonword reading, colour naming and picture naming. Repetitive transcranial magnetic stimulation (rTMS) was delivered over posterior BA37 of the left and right hemispheres (lBA37 and rBA37, respectively) and over the vertex. Subjects were significantly slower to name pictures when TMS was given over lBA37 compared to vertex or rBA37. rTMS over lBA37 had no significant effect on word reading, nonword reading or colour naming. The picture naming deficit is suggested to result from a disruption to object recognition processes. This study corroborates the finding from a recent imaging study, that the most posterior part of left hemispheric BA37 has a necessary role in object recognition.


Manipulating directional cell motility using intracellular superparamagnetic nanoparticles.
This study investigated the ability for magnetic nanoparticles to influence cellular migration in the presence of an external magnetic field. We found that the direction of migrating keratinocytes can be controlled and the migration speed of fibroblasts can be increased with the internalisation of these nanoparticles in the presence of a magnetic field. The possibility of shepherding cells towards a region of interest through the use of internalized nanoparticles is an attractive prospect for cell tracking, cell therapies, and tissue engineering applications.


Mucoadhesive Bletilla striata Polysaccharide-Based Artificial Tears to Relieve Symptoms and Inflammation in Rabbit with Dry Eyes Syndrome.
Dry eye syndrome (DES) is a multifactorial disorder of the ocular surface affecting many people all over the world. However, there have been many therapeutic advancements for the treatment of DES, substantial long-term treatment remains a challenge. Natural plant-based polysaccharides have gained much importance in the field of tissue engineering for their excellent biocompatibility and unique physical properties. In this study, polysaccharides from a Chinese ground orchid, Bletilla striata, were successfully extracted and incorporated into the artificial tears for DES treatment due to its anti-inflammatory and mucoadhesive properties. The examination for physical properties such as refractive index, pH, viscosity and osmolality of the Bletilla striata polysaccharide (BSP) artificial tears fabricated in this study showed that it was in close association with that of the natural human tears. The reactive oxygen species (ROS) level and inflammatory gene expression tested in human corneal epithelium cells (HCECs) indicated that the low BSP concentrations (0.01-0.1% v/v) could effectively reduce inflammatory cytokines (TNF, IL8) and ROS levels in HCECs, respectively. Longer retention of the BSP-formulated artificial tears on the ocular surface is due to the mucoadhesive nature of BSP allowing lasting lubrication. Additionally, a rabbit's DES model was created to evaluate the effect of BSP for treating dry eye. Schirmer test results exhibited the effectiveness of 0.1% (v/v) BSP-containing artificial tears in enhancing the tear volume in DES rabbits. This work combines the effectiveness of artificial tears and anti-inflammatory herb extract (BSP) to moisturize ocular surface and to relieve the inflammatory condition in DES rabbit, which further shows great potential of BSP in treating ocular surface diseases like DES in clinics in the future.


Interaction of syntaxin with a single Kv1.1 channel: a possible mechanism for modulating neuronal excitability.
Voltage-gated K(+) channels are crucial for intrinsic neuronal plasticity and present a target for modulations by protein-protein interactions, notably, by exocytotic proteins demonstrated by us in several systems. Here, we investigated the interaction of a single Kv1.1 channel with syntaxin 1A. Syntaxin decreased the unitary conductance of all conductance states (two subconductances and a full conductance) and decreased their open probabilities by prolongation of mean closed dwell-times at depolarized potentials. However, at subthreshold potentials syntaxin 1A increased the probabilities of the subconductance states. Consequently, the macroscopic conductance is decreased at potentials above threshold and increased at threshold potentials. Numerical modeling based on steady-state and kinetic analyses suggests: (1) a mechanism whereby syntaxin controls activation gating by forcing the conductance pathway only via a sequence of discrete steps through the subconductance states, possibly via a breakdown of cooperative movements of voltage sensors that exist in Kv1.1; (2) a physiological effect, apparently paradoxical for an agent that reduces K(+) current, of attenuating neuronal firing frequency via an increase in K(+) shunting conductance. Such modulation of the gain of neuronal output in response to different levels of syntaxin is in accord with the suggested role for Kv1.1 in axonal excitability and synaptic efficacy.


Effectiveness of Social Skills Interventions Incorporating Peer Interactions for Children With Attention Deficit Hyperactivity Disorder: A Systematic Review.
Few studies examining the use of peers during interventions have been published, and no systematic review has been conducted to evaluate the available literature. To examine the effectiveness of social skills interventions incorporating peers for children with attention deficit hyperactivity disorder (ADHD) to improve social interactions. A search of five databases (CINAHL, PubMed, Web of Science, Google Scholar, and PsycINFO) produced 697 articles. Sixty-one were retrieved for full-text review, and 15 articles met inclusion criteria. Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines were used to abstract data. Inclusion criteria: Participants younger than age 18 yr with any ADHD pattern, social skills interventions with peer involvement, outcome measures within the domain of occupational therapy, written in English, and involved a peer as the sole or primary component at some point in the social skills intervention. Exclusion criteria: Studies older than 20 yr or that used participants with comorbidities or multiple conditions. Interventions incorporating both peer categories were effective for increasing play skills, reducing undesirable social behaviors (e.g., inappropriate verbalizations, dominant behaviors, aggression), and improving communication (e.g., pragmatic language, collaboration, joint participation) and social participation. Improvements were maintained over time, as evidenced by follow-up studies. Outcomes of these studies demonstrate moderate evidence that supports the use of social skills interventions incorporating peers for children with ADHD to improve social interactions, supporting their use by occupational therapists and the need for more studies. This article provides guidance to occupational therapy practitioners on social skills intervention options for children with ADHD.


DISTINCT MECHANISMS OF INHIBITION OF VSV REPLICATION IN NEURONS MEDIATED BY TYPE I AND TYPE II IFN.
Acute viral infection of neurons presents a difficult problem to the host, since neurons are essential and not replaced, therefore cell-autonomous pathway(s) of suppressing viral replication are critical. We have examined the mechanisms by which neurons respond to exogenous interferons (IFNs) and observed that novel pathways inhibit acute vesicular stomatitis virus (VSV) replication. For both type I (IFN-beta) and Type II (IFN-gamma) interferons, post-translational modification of viral proteins contributed to the replication blockade, diminishing the efficiency of viral assembly and budding from the host neuron. IFN-gamma treatment induces the accumulation of NOS-1 in the absence of an increase of mRNA encoding this enzyme; a NOS-1-inhibiting protein, PIN, is rapidly ubiquitinated and eliminated in the presence of IFN-gamma. NOS-1 produces NO which combines with superoxide to form peroxynitrite (ONOO-), this binds tyrosines, cysteines, and serines; antagonism of NOS-1 with either non-specific or selective inhibitors block the antiviral effect of IFN-gamma. VSV proteins are decorated with -NO(2) in IFN-gamma-treated neurons, probably resulting in their diminished ability to interact properly and mature into budding virus. For IFN-beta, protein phosphorylation of the Matrix protein (M) and Phosphoprotein (P) were altered in infected neurons, with hyperphosphorylation of M (but not hypophosphorylated P) found in released virions. Hyperphosphorylated M protein does not immunoprecipitate with the viral ribonucleoprotein complex in IFN-beta-treated neurons. Thus both types of IFN interfere with viral assembly and release of infectious particles, but by distinct pathways.


Transgenic mice expressing an intracellular fluorescent fusion of angiotensin II demonstrate renal thrombotic microangiopathy and elevated blood pressure.
We have generated transgenic mice that express angiotensin II (ANG II) fused downstream of enhanced cyan fluorescent protein, expression of which is regulated by the mouse metallothionein promoter. The fusion protein, which lacks a secretory signal, is retained intracellularly. In the present study, RT-PCR, immunoblot analyses, whole-animal fluorescent imaging, and fluorescent microscopy of murine embryonic fibroblasts confirm expression of the fusion protein in vivo and in vitro. The transgene is expressed in all tissues tested (including brain, heart, kidney, liver, lung, and testes), and radioimmunoassay of plasma samples obtained from transgenic mice indicate no increase in circulating ANG II over wild-type levels, consistent with intracellular retention of the transgene product. Kidneys from transgenic and corresponding wild-type littermates were histologically evaluated, and abnormalities in transgenic mice consistent with thrombotic microangiopathy were observed; microthrombosis was frequently observed within the glomerular capillaries and small vessels. In addition, systolic and diastolic blood pressures, measured by telemetry (n = 8 for each group), were significantly higher in transgenic mice compared with wild-type littermates. Blood pressure of line A male transgenic mice was 125 + or - 1.7 over 97 + or - 1.6 compared with 109 + or - 1.7 over 83 + or - 1.4 mmHg in wild-type littermates (systolic over diastolic). In summary, overexpression of an intracellular fluorescent fusion protein of ANG II correlates with elevated blood pressure and kidney pathology. This transgenic model may be useful to further explore the intracellular renin-angiotensin system and its implication in abnormal kidney function and hypertension.


Effects of Exercise on Liver Fat and Metabolism in Alcohol Drinkers.
Exercise is an important component of obesity-associated disorders and has been shown to reduce markers of nonalcoholic fatty liver disease (NAFLD). However, little is known about how these effects are influenced by alcohol intake. The authors performed a randomized controlled trial to investigate the effects of exercise on hepatic triglyceride content (HTGC) and metabolism in overweight or obese patients who consume alcohol. The authors performed a prospective study of 27 patients (mean 54 ± 11 years of age, body mass index [BMI] 31 ± 4 kg/m2) with >5% HTGC in the United Kingdom, consuming alcohol (mean 221 ± 75 g/week). Anthropometry, body composition, HTGC, and abdominal fat were measured using plethysmography and magnetic resonance imaging. Subjects were assigned to groups that exercised (3 times/week on nonconsecutive days) for 12 weeks (n = 14) or continued standard care (control group, n = 13), maintaining baseline weight and alcohol consumption. The exercise program consisted of aerobic exercise (static cycling) and a circuit of resistance exercise (free weights and machines). Patients were examined at baseline and at 12 weeks; data collected on HTGC, body composition, metabolic control, circulating inflammatory, and fibrosis markers were assessed at baseline and at 12 weeks. Between-group differences were evaluated using an unpaired t test and within-group differences using a paired t test. The primary outcomes for this study were changes in HTGC between baseline and 12 weeks. After 12 weeks, there was no significant difference between the exercise and control groups in HTGC (reduction of 0.1% ± 2.1% in exercisers vs increase of 0.5 ± 2.1% in control group; P > .05). At week 12, the exercise group had significant reductions in subcutaneous fat (loss of 23 ± 28 cm2 in the exercisers vs increase of 12 ± 19 cm2 in the control group; P < .01), and whole body fat (loss of 2.1 ± 1.1 kg in the exercisers vs increase of 0.2 ± 2.1 kg; P < .01). The exercise group also had a significantly greater increase in lean body mass (increase of 1.9 ± 1.4 kg for the exercisers vs increase of 0.7 ± 1.5 kg for the control group; P < .01) and a significantly greater reduction in level of cytokeratin 18 (reduction of 49 ± 82 U/L in exercisers vs increase of 17 ± 38 U/L in control group; P < .05). There were no differences between groups in changes in metabolic factors or markers of inflammation. In a randomized controlled trial of obese individuals who consume alcohol, exercise significantly improved body composition and reduced hepatocyte apoptosis (cytokeratin 18), but did not reduce HTGC. This finding could indicate that alcohol consumption reduces the effects of exercise on NAFLD observed in previous studies. Clinical care teams should look to use exercise as part of the management strategy for people consuming alcohol, but optimal benefit may be as an adjunct to alcohol reduction and weight management strategies. (ISRCTN.com, Number: ISRCTN90597099).


The cost effectiveness of strategies for the treatment of intestinal parasites in immigrants.
Currently, more than 600,000 immigrants enter the United States each year from countries where intestinal parasites are endemic. At entry persons with parasitic infections may be asymptomatic, and stool examinations are not a sensitive method of screening for parasitosis. Albendazole is a new, broad-spectrum antiparasitic drug, which was approved recently by the Food and Drug Administration. International trials have shown albendazole to be safe and effective in eradicating many parasites. In the United States there is now disagreement about whether to screen all immigrants for parasites, treat all immigrants presumptively, or do nothing unless they have symptoms. We compared the costs and benefits of no preventive intervention (watchful waiting) with those of universal screening or presumptive treatment with 400 mg of albendazole per day for five days. Those at risk were defined as immigrants to the United States from Asia, the Middle East, sub-Saharan Africa, Eastern Europe, and Latin America and the Caribbean. Cost effectiveness was expressed both in terms of the cost of treatment per disability-adjusted life-year (DALY) averted (one DALY is defined as the loss of one year of healthy life to disease) and in terms of the cost per hospitalization averted. As compared with watchful waiting, presumptive treatment of all immigrants at risk for parasitosis would avert at least 870 DALYs, prevent at least 33 deaths and 374 hospitalizations, and save at least $4.2 million per year. As compared with watchful waiting, screening would cost $159,236 per DALY averted. Presumptive administration of albendazole to all immigrants at risk for parasitosis would save lives and money. Universal screening, with treatment of persons with positive stool examinations, would save lives but is less cost effective than presumptive treatment.


Assessing vigilance in caregivers after traumatic brain injury: TBI-CareQOL Caregiver Vigilance.
Caregivers of individuals with traumatic brain injury (TBI) frequently experience anxiety related to the caregiver role. Often this is due to a caregiver's perceived need to avoid people and situations that might upset or "trigger" the care recipient. There are currently no self-report measures that capture these feelings; thus, this article describes the development and preliminary validation efforts for the TBI-Caregiver Quality of Life (CareQOL) Caregiver Vigilance item bank. A sample of 532 caregivers of civilians (n = 218) or service members/veterans (SMVs; n = 314) with TBI completed 32 caregiver vigilance items, other measures of health-related quality of life (RAND-12, Patient-Reported Outcomes Measurement Information System [PROMIS] Depression, PROMIS Social Isolation, Caregiver Appraisal Scale), and the Mayo-Portland Adaptability Inventory-4. The final item bank contains 18 items, as supported by exploratory and confirmatory factor analysis, item response theory graded response modeling (GRM), and differential item functioning investigations. Expert review and GRM calibration data informed the selection of a 6-item short form and programming of a computer adaptive test. Internal consistency reliability for the different administration formats were excellent (reliability coefficients ≥ .90). Three-week test-retest stability was supported (i.e., r ≥ .78). Correlations between vigilance and other self-report measures supported convergent and discriminant validity (0.01 ≤ r ≤ .69). Known-groups validity was also supported. The new TBI-CareQOL Caregiver Vigilance computer adaptive test and corresponding 6-item short form were developed using established rigorous measurement development standards, providing the first self-report measure to evaluate caregiver vigilance. This development work indicates that this measure exhibits strong psychometric properties. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


Microbially enhanced dissolution of HgS in an acid mine drainage system in the California Coast Range.
Mercury sulfides (cinnabar and metacinnabar) are the main ores of Hg and are relatively stable under oxic conditions (Ksp = 10⁻⁵⁴ and 10⁻⁵² , respectively). However, until now their stability in the presence of micro-organisms inhabiting acid mine drainage (AMD) systems was unknown. We tested the effects of the AMD microbial community from the inoperative Hg mine at New Idria, CA, present in sediments of an AMD settling pond adjacent to the main waste pile and in a microbial biofilm on the surface of this pond, on the solubility of crystalline HgS. A 16S rRNA gene clone library revealed that the AMD microbial community was dominated by Fe-oxidizing (orders Ferritrophicales and Gallionellas) and S-oxidizing bacteria (Thiomonas sp.), with smaller amounts (≤ 6%) being comprised of the orders Xanthomondales and Rhodospirillales. Though the order Ferritrophicales dominate the 16S rRNA clones (>60%), qPCR results of the microbial community indicate that the Thiomonas sp. represents ~55% of the total micro-organisms in the top 1 cm of the AMD microbial community. Although supersaturated with respect to cinnabar and metacinnabar, microcosms inoculated with the AMD microbial community were capable of releasing significantly more Hg into solution compared to inactivated or abiotic controls. Four different Hg-containing materials were tested for bacterially enhanced HgS dissolution: pure cinnabar, pure metacinnabar, mine tailings, and calcine material (processed ore). In the microcosm with metacinnabar, the presence of the AMD microbial community resulted in an increase of dissolved Hg concentrations up to 500 μg L ⁻¹during the first 30 days of incubation. In abiotic control microcosms, dissolved Hg concentrations did not increase above 100 ng L⁻¹ . When Hg concentrations were below 50 μg L⁻¹ , the Fe-oxidizing bacteria in the AMD microbial community were still capable of oxidizing Fe(II) to Fe(III) in the AMD solution, whereas concentrations above 50 μg L⁻¹ resulted in inhibition of microbial iron oxidation. Our experiments show that the AMD microbial community contributes to the dissolution of mercury sulfide minerals. These findings have major implications for risk assessment and future management of inoperative Hg mines worldwide.


Determining the Agreement Between an Automated Respiratory Rate Counter and a Reference Standard for Detecting Symptoms of Pneumonia in Children: Protocol for a Cross-Sectional Study in Ethiopia.
Acute respiratory infections (ARIs), primarily pneumonia, are the leading infectious cause of under-5 mortality worldwide. Manually counting respiratory rate (RR) for 60 seconds using an ARI timer is commonly practiced by community health workers to detect fast breathing, an important sign of pneumonia. However, correctly counting breaths manually and classifying the RR is challenging, often leading to inappropriate treatment. A potential solution is to introduce RR counters, which count and classify RR automatically. This study aims to determine how the RR count of an Automated Respiratory Infection Diagnostic Aid (ARIDA) agrees with the count of an expert panel of pediatricians counting RR by reviewing a video of the child's chest for 60 seconds (reference standard), for children aged younger than 5 years with cough and/or difficult breathing. A cross-sectional study aiming to enroll 290 children aged 0 to 59 months presenting to pediatric in- and outpatient departments at a teaching hospital in Addis Ababa, Ethiopia, was conducted. Enrollment occurred between April and May 2017. Once enrolled, children participated in at least one of three types of RR evaluations: (1) agreement-measure the RR count of an ARIDA in comparison with the reference standard, (2) consistency-measure the agreement between two ARIDA devices strapped to one child, and (3) RR fluctuation-measure RR count variability over time after ARIDA attachment as measured by a manual count. The agreement and consistency of expert clinicians (ECs) counting RR for the same child with the Mark 2 ARI timer for 60 seconds was also measured in comparison with the reference standard. Primary outcomes were (1) mean difference between the ARIDA and reference standard RR count (agreement) and (2) mean difference between RR counts obtained by two ARIDA devices started simultaneously (consistency). Study strengths included the design allowing for comparison between both ARIDA and the EC with the reference standard RR count. A limitation is that exactly the same set of breaths were not compared between ARIDA and the reference standard since ARIDA can take longer than 60 seconds to count RR. Also, manual RR counting, even when aided by a video of the child's chest movements, is subject to human error and can result in low interrater reliability. Further work is needed to reach global consensus on the most appropriate reference standard and an acceptable level of agreement to provide ministries of health with evidence to make an informed decision on whether to scale up new automated RR counters. ClinicalTrials.gov NCT03067558; https://clinicaltrials.gov/ct2/show/NCT03067558. RR1-10.2196/16531.


Design of renewable hydrogel release systems from fiberboard mill wastewater.
A new route for the design of renewable hydrogels is presented. The soluble waste from masonite production was isolated, fractionized, and upgraded. The resulting hemicellulose rich fraction was alkenyl-functionalized and used in the preparation of covalently cross-linked hydrogels capable of sustained release of incorporated agents. Said hydrogels showed a Fickian diffusion-based release of incorporated bovine serum albumin. Also, a method for the coating of seeds with hydrogel was developed. The sustained release of incorporated growth retardant agents from the hydrogel coating on rape seeds was shown to enable the temporary inhibition of germination.


Influence of soil properties on the bioaccessibility of Cr and Ni in geologic serpentine and anthropogenically contaminated non-serpentine soils in Taiwan.
Serpentine minerals with high levels of geologic chromium (Cr) and nickel (Ni) and non-serpentine farmlands polluted by irrigation water causing high anthropogenic Cr and Ni levels are both found in Taiwan. Elevated levels of Cr and Ni in these soils are a concern due to their potential to promote cancer mortality in humans. Bioaccessibility is a crucial factor determining the actual health risk via oral ingestion when children are exposed to metal-contaminated soils. Furthermore, the bioaccessibility of metals varies with the source, soil properties, and fractionation of metals in the soil. Therefore in this study, soil pH, total organic carbon (TOC), texture, and the total concentrations, fractionation, and bioaccessibility of Cr and Ni were analyzed and correlated for soils collected from serpentine mineral-containing deposits and contaminated non-serpentine farmlands. The low bioaccessibility and low mobility of Cr and Ni in serpentine soils suggested that incidental ingesting of soils posed a low health risk; however, the higher bioaccessibility and mobility of Ni in non-serpentine soils contaminated by electroplating wastewater could lead to potential risks for humans. Additionally, a significant difference in the bioaccessibility of Ni was observed between serpentine and non-serpentine soils, but this was not shown for Cr. Accordingly, a correlation analysis showed that Cr bioaccessibility was positively correlated with TOC, with no distinction between serpentine and non-serpentine soils. In contrast, TOC and the fractions of the sequential extraction procedure were significantly correlated with Ni bioaccessibility both in anthropogenically contaminated non-serpentine soils and in natural serpentine soils.


Paper-based and web-based intervention modeling experiments identified the same predictors of general practitioners' antibiotic-prescribing behavior.
To evaluate the robustness of the intervention modeling experiment (IME) methodology as a way of developing and testing behavioral change interventions before a full-scale trial by replicating an earlier paper-based IME. Web-based questionnaire and clinical scenario study. General practitioners across Scotland were invited to complete the questionnaire and scenarios, which were then used to identify predictors of antibiotic-prescribing behavior. These predictors were compared with the predictors identified in an earlier paper-based IME and used to develop a new intervention. Two hundred seventy general practitioners completed the questionnaires and scenarios. The constructs that predicted simulated behavior and intention were attitude, perceived behavioral control, risk perception/anticipated consequences, and self-efficacy, which match the targets identified in the earlier paper-based IME. The choice of persuasive communication as an intervention in the earlier IME was also confirmed. Additionally, a new intervention, an action plan, was developed. A web-based IME replicated the findings of an earlier paper-based IME, which provides confidence in the IME methodology. The interventions will now be evaluated in the next stage of the IME, a web-based randomized controlled trial.


Stimulation of the hypothalamic ventromedial nuclei by pituitary adenylate cyclase-activating polypeptide induces hypophagia and thermogenesis.
Numerous studies have demonstrated that the hypothalamic ventromedial nuclei (VMN) regulate energy homeostasis by integrating and utilizing behavioral and metabolic mechanisms. The VMN heavily express pituitary adenylate cyclase-activating polypeptide (PACAP) type I receptors (PAC1R). Despite the receptor distribution, most PACAP experiments investigating affects on feeding have focused on intracerebroventricular administration or global knockout mice. To identify the specific contribution of PACAP signaling in the VMN, we injected PACAP directly into the VMN and measured feeding behavior and indices of energy expenditure. Following an acute injection of PACAP, nocturnal food intake was significantly reduced for 6 h after injections without evidence of malaise. In addition, PACAP-induced suppression of feeding also occurred following an overnight fast and could be blocked by a specific PAC1R antagonist. Metabolically, VMN-specific injections of PACAP significantly increased both core body temperature and spontaneous locomotor activity with a concurrent increase in brown adipose uncoupling protein 1 mRNA expression. To determine which signaling pathways were responsive to PACAP administration into the VMN, we measured mRNA expression of well-characterized hypothalamic neuropeptide regulators of feeding. One hour after PACAP administration, expression of pro-opiomelanocortin mRNA was significantly increased in the arcuate nuclei (ARC), with no changes in neuropeptide Y and agouti-related polypeptide mRNA levels. This suggests that PAC1R expressing VMN neurons projecting to pro-opiomelanocortin neurons contribute to hypophagia by involving melanocortin signaling. While the VMN also abundantly express PACAP protein, the present study demonstrates that PACAP input to the VMN can influence the control of energy homeostasis.


STING agonist formulated cancer vaccines can cure established tumors resistant to PD-1 blockade.
Stimulator of interferon genes (STING) is a cytosolic receptor that senses both exogenous and endogenous cytosolic cyclic dinucleotides (CDNs), activating TBK1/IRF3 (interferon regulatory factor 3), NF-κB (nuclear factor κB), and STAT6 (signal transducer and activator of transcription 6) signaling pathways to induce robust type I interferon and proinflammatory cytokine responses. CDN ligands were formulated with granulocyte-macrophage colony-stimulating factor (GM-CSF)-producing cellular cancer vaccines--termed STINGVAX--that demonstrated potent in vivo antitumor efficacy in multiple therapeutic models of established cancer. We found that rationally designed synthetic CDN derivative molecules, including one with an Rp,Rp dithio diastereomer and noncanonical c[A(2',5')pA(3',5')p] phosphate bridge structure, enhanced antitumor efficacy of STINGVAX in multiple aggressive therapeutic models of established cancer in mice. Antitumor activity was STING-dependent and correlated with increased activation of dendritic cells and tumor antigen-specific CD8(+) T cells. Tumors from STINGVAX-treated mice demonstrated marked PD-L1 (programmed death ligand 1) up-regulation, which was associated with tumor-infiltrating CD8(+)IFNγ(+) T cells. When combined with PD-1 (programmed death 1) blockade, STINGVAX induced regression of palpable, poorly immunogenic tumors that did not respond to PD-1 blockade alone.


Effects of feeding omega-3-fatty acids on fatty acid composition and quality of bovine sperm and on antioxidative capacity of bovine seminal plasma.
The aim of the present study was to examine the effects of feeding alpha-linolenic (ALA) acid on fatty acid composition and quality of bovine sperm and on antioxidative capacity of seminal plasma. Nine bulls (ALA bulls) were fed with 800 g rumen-resistant linseed oil with a content of 50% linolenic acid and eight bulls with 400 g palmitic acid (PA bulls). Sperm quality was evaluated for plasma membrane and acrosome intact sperm (PMAI), the amount of membrane lipid peroxidation (LPO), and the percentage of sperm with a high DNA fragmentation index (DFI). Fatty acid content of sperm was determined using gas chromatography. Total antioxidant capacity, glutathione peroxidase, and superoxide dismutase activity were determined in seminal plasma. Feeding ALA increased (P < 0.05) the docosahexaenoic acid (DHA) content in bulls whereas in PA bulls did not change. PMAI increased after cryopreservation in ALA bulls as well as in PA bulls during the experiment period (P < 0.005). LPO of sperm directly after thawing did not change during the study period in ALA group, but decreased in PA group (P < 0.006). After 3h of incubation LPO increased in the ALA group (P < 0.02), while LPO did not differ between phases within groups. In conclusion, feeding of neither saturated nor polyunsaturated fatty acids affect the antioxidant levels in seminal plasma. Both saturated as well as polyunsaturated fatty acids had positive effects on quality of cryopreserved bovine sperm, although the content of docosahexaenoic acid in sperm membranes increased only in ALA bulls.


Validity and user-friendliness of the minimal eating observation and nutrition form - version II (MEONF - II) for undernutrition risk screening.
To analyze the criterion-related validity and user-friendliness of the Minimal Eating Observation and Nutrition Form - Version II (MEONF - II) and Malnutrition Universal Screening Tool (MUST) in relation to the Mini Nutritional Assessment (MNA). In addition, the effect of substituting body mass index (BMI) with calf circumference (CC) was explored for the MEONF-II. The study included 100 patients who were assessed for nutritional status with the MNA (full version), considered here to be the gold standard, and screened with the MUST and the MEONF-II. The MEONF-II includes assessments of involuntary weight loss, BMI (or calf circumference), eating difficulties, and presence of clinical signs of undernutrition. The MEONF-II sensitivity (0.73) and specificity (0.88) were acceptable. Sensitivity and specificity for the MUST were 0.57 and 0.93, respectively. Replacing the BMI with CC in the MEONF-II gave similar results (sensitivity 0.68, specificity 0.90). Assessors considered MEONF-II instructions and items to be relevant, easy to understand and complete (100%), and the questions to be relevant (98%). MEONF-II and MUST took 8.8 and 4.7 minutes to complete, respectively, and both were considered relevant and easy to finish. In addition, MEONF-II was thought to reveal problems that allows for nursing interventions. The MEONF-II is an easy to use, relatively quick, and sensitive screening tool to assess risk of undernutrition among hospital inpatients, which allows for substituting BMI with CC in situations where measures of patient height and weight cannot be easily obtained. High sensitivity is of primary concern in nutritional screening and the MEONF-II outperforms the MUST in this regard.


Glucocorticoid and mineralocorticoid receptors and associated diseases.
Adrenal corticosteroids, i.e. glucocorticoids and mineralocorticoids, play important physiological roles in humans. Their actions are mediated by intracellular receptor molecules, the glucocorticoid receptor (GR) and mineralocorticoid receptor (MR), which function as hormone-dependent transcription factors. Ligand-activated receptors modulate the transcription rates of responsive genes by interacting with responsive elements in the promoters of these genes or by influencing the activities of other transcription factors, via protein-protein interactions. Natural inactivating mutations of the GR or MR genes have been reported in humans with significant clinical phenotypes. The former causes sporadic or familial glucocorticoid resistance characterized by generalized partial insensitivity of tissues to glucocorticoids and subsequent activation of the hypothalamic/pituitary/adrenal axis with resultant hyperandrogenism in children and women and/or mineralocorticoid excess symptoms in both sexes. The latter develop pseudohypoaldosteronism type 1, i.e. hypotension and hyperkalaemic acidosis, as a result of reduced aldosterone actions in the kidney. An activating mutation in the MR gene causing early-onset, periodic hypertension was reported recently. The biological relevance of the GR and MR receptors was also addressed in mice whose GR or MR genes were inactivated or modified by gene targeting. The results were generally confirmatory of the concepts obtained by the human studies. Similarly, natural, compensated glucocorticoid and/or mineralocorticoid 'resistance' were described in several mammalian species, including non-human primates and rodents. Here we discuss the actions of GR and MR and the molecular defects of naturally occuring mutations in these receptors with associated pathophysiological changes.


Plants as a source of butyrylcholinesterase variants designed for enhanced cocaine hydrolase activity.
Cocaine addiction affects millions of people with disastrous personal and social consequences. Cocaine is one of the most reinforcing of all drugs of abuse, and even those who undergo rehabilitation and experience long periods of abstinence have more than 80% chance of relapse. Yet there is no FDA-approved treatment to decrease the likelihood of relapse in rehabilitated addicts. Recent studies, however, have demonstrated a promising potential treatment option with the help of the serum enzyme butyrylcholinesterase (BChE), which is capable of breaking down naturally occurring (-)-cocaine before the drug can influence the reward centers of the brain or affect other areas of the body. This activity of wild-type (WT) BChE, however, is relatively low. This prompted the design of variants of BChE which exhibit significantly improved catalytic activity against (-)-cocaine. Plants are a promising means to produce large amounts of these cocaine hydrolase variants of BChE, cheaply, safely with no concerns regarding human pathogens and functionally equivalent to enzymes derived from other sources. Here, in expressing cocaine-hydrolyzing mutants of BChE in Nicotiana benthamiana using the MagnICON virus-assisted transient expression system, and in reporting their initial biochemical analysis, we provide proof-of-principle that plants can express engineered BChE proteins with desired properties.


Improved electron-hole separation and migration in anatase TiO2 nanorod/reduced graphene oxide composites and their influence on photocatalytic performance.
The as-synthesized TiO2 nanorods a-TNR (amorphous TiO2 layer covering the crystalline anatase TiO2 core) and TNR (fully crystalline anatase TiO2) were decorated with reduced graphene oxide (rGO) to synthesize two series of TiO2 + rGO composites with different nominal loadings of GO (from 4 to 20 wt%). The structural, surface and electronic properties of the obtained TiO2 + rGO composites were analyzed and correlated to their performance in the photocatalytic oxidation of aqueous bisphenol A solution. X-ray photoelectron spectroscopy (XPS) analyses revealed that charge separation in TiO2 + rGO composites is improved due to the perfect matching of TiO2 and rGO valence band maxima (VBM). Cyclic voltammetry (CV) experiments revealed that the peak-to-peak separations (ΔEp) are the lowest and the oxidation current densities are the highest for composites with a nominal 10 wt% GO content, meaning that it is much easier for the charge carriers to percolate through the solid, resulting in improved charge migration. Due to the high charge carrier mobility in rGO and perfect VBM matching between TiO2 and rGO, the electron-hole recombination in composites was suppressed, resulting in more electrons and holes being able to participate in the photocatalytic reaction. rGO amounts above 10 wt% decreased the photocatalytic activity; thus, it is critical to optimize its amount in the TiO2 + rGO composites for achieving the highest photocatalytic activity. BPA degradation rates correlated completely with the results of the CV measurements, which directly evidenced improved charge separation and migration as the crucial parameters governing photocatalysis.


Biotin status assessed longitudinally in pregnant women.
This study assessed biotin nutritional status longitudinally during pregnancy as judged by urinary excretion of biotin and biotin metabolites and by serum concentration of biotin. 3-Hydroxyisovaleric acid excretion was also assessed because increased excretion of that acid reflects decreased tissue activity of the biotin-dependent enzyme, methylcrotonyl-CoA carboxylase. Thirteen women provided untimed urine samples during both early and late pregnancy. Twelve nonpregnant women served as controls. Biotin and metabolites were determined by a combined HPLC/avidin-binding assay. 3-Hydroxyisovaleric acid was determined by gas chromatography/mass spectrophotometry. Significance of changes from early to late pregnancy was tested by paired t test; to compare nonpregnant controls with early and late pregnancy, ANOVA was used. During early pregnancy, biotin excretion was not significantly different than controls; however, 3-hydroxyisovaleric acid excretion was significantly increased relative to controls (P < 0.0001) and was greater than the upper limit of normal in 9 of 13 women. From early to late pregnancy, biotin excretion decreased in 10 of 13 women (P < 0.01); by late pregnancy, biotin excretion was less than normal in six women. During late pregnancy, 3-hydroxyisovaleric acid remained significantly increased relative to controls (P < 0.0001). Serum concentrations of biotin were significantly greater than those of controls during early pregnancy (P < 0.0001) and decreased in each woman from early to late pregnancy (P < 0.0001). These data provide evidence that biotin status decreases during pregnancy.


Strong association between west african rainfall and u.s. Landfall of intense hurricanes.
Intense hurricanes occurred much more frequently during the period spanning the late 1940s through the late 1960s than during the 1970s and 1980s, except for 1988 and 1989. Seasonal and multidecadal variations of intense hurricane activity are closely linked to seasonal and multidecadal variations of summer rainfall amounts in the Western Sahel region of West Africa. The multidecadal nature of West African precipitation variations and their association with variations of intense Atlantic hurricane activity can be observed in data going back nearly a century. The apparent recent breaking of the 18-year Sahel drought during 1988 and 1989 suggests that the incidence of intense hurricanes making landfall on the U.S. coast and in the Caribbean basin will likely increase during the 1990s and early years of the 21st century to levels of activity notably greater than were observed during the 1970s and 1980s.


The effect of plasmon field on the coherent lattice phonon oscillation in electron-beam fabricated gold nanoparticle pairs.
By using electron beam lithography, we fabricated pairs of gold nanoparticles with varying interparticle separation. Double-beam femtosecond transient absorption spectroscopy was used to determine the coherent lattice oscillation frequency as a function of the interparticle separation in the presence of the plasmon field excited by the monitoring probe light. We found that the fractional shift in the coherent lattice phonon oscillation frequency follows an exponential decay with respect to the interparticle gap scaled by the disc diameter with the same decay constant as that previously observed for the fractional shift in the surface plasmon electronic oscillation resonance frequency. This strongly suggests that it is the near-field coupling between the particles that shifts both the coherent electronic oscillation (plasmon) frequency and the coherent lattice oscillation (phonon) frequency. The similar trend in the effect of interparticle coupling on the plasmon frequency and the phonon frequency is essentially a reflection of the universal scaling behavior of the distance decay of the interparticle plasmonic near-field. It is shown that the observed decrease in the lattice oscillation frequency with decrease in the interparticle distance is the result of a reduction in the effective free electron density within each nanoparticle pair partner as a result of the polarizing perturbation of the plasmonic field of the other nanoparticle in the pair.


Experimental and Theoretical Analysis of Nanotransport in Oligophenylene Dithiol Junctions as a Function of Molecular Length and Contact Work Function.
We report the results of an extensive investigation of metal-molecule-metal tunnel junctions based on oligophenylene dithiols (OPDs) bound to several types of electrodes (M1-S-(C6H4)n-S-M2, with 1 ≤ n ≤ 4 and M1,2 = Ag, Au, Pt) to examine the impact of molecular length (n) and metal work function (Φ) on junction properties. Our investigation includes (1) measurements by scanning Kelvin probe microscopy of electrode work function changes (ΔΦ = ΦSAM - Φ) caused by chemisorption of OPD self-assembled monolayers (SAMs), (2) measurements of junction current-voltage (I-V) characteristics by conducting probe atomic force microscopy in the linear and nonlinear bias ranges, and (3) direct quantitative analysis of the full I-V curves. Further, we employ transition voltage spectroscopy (TVS) to estimate the energetic alignment εh = EF - EHOMO of the dominant molecular orbital (HOMO) relative to the Fermi energy EF of the junction. Where photoelectron spectroscopy data are available, the εh values agree very well with those determined by TVS. Using a single-level model, which we justify via ab initio quantum chemical calculations at post-density functional theory level and additional UV-visible absorption measurements, we are able to quantitatively reproduce the I-V measurements in the whole bias range investigated (∼1.0-1.5 V) and to understand the behavior of εh and Γ (contact coupling strength) extracted from experiment. We find that Fermi level pinning induced by the strong dipole of the metal-S bond causes a significant shift of the HOMO energy of an adsorbed molecule, resulting in εh exhibiting a weak dependence with the work function Φ. Both of these parameters play a key role in determining the tunneling attenuation factor (β) and junction resistance (R). Correlation among Φ, ΔΦ, R, transition voltage (Vt), and εh and accurate simulation provide a remarkably complete picture of tunneling transport in these prototypical molecular junctions.


Older Age as a Prognostic Factor of Attenuated Pain Recovery After Shoulder Arthroscopy.
Shoulder pain and surgery are common among older adults. However, the extent to which older age affects recovery after shoulder surgery is not well understood. To assess influence of older age on postoperative recovery factors 3 and 6 months after shoulder arthroscopy. Prospective cohort study. University-affiliated outpatient orthopedic surgical center. A convenience sample of 139 persons between 20 and 79 years of age who experienced shoulder pain, had musculoskeletal dysfunction based on imaging and physician assessment, and were scheduled for an arthroscopic shoulder procedure. Postoperative outcomes were compared among younger, middle-aged, and older adults before surgery and at 3 and 6 months after surgery using analysis of variance modeling. Movement-evoked pain and an experimental laboratory correlate of pain processing were assessed at each time point. The influence of older age on 3- and 6-month pain outcomes were determined via multivariate regression analyses after accounting for preoperative, intraoperative, and postoperative prognostic factors. Older adults had higher movement-evoked pain intensity (F2,108 = 5.18, P = .007) and experimental pain response (F2,111 = 7.24, P = .001) at 3 months compared with young and middle-aged adults. After controlling for key prognostic factors, older age remained a positive predictor of 3-month movement-evoked pain (R(2) = 0.05; standardized [St.] β = 0.263, P = .031) and experimental pain response (R(2) = 0.07; St. β = 0.295, P = .014). Further, older age remained a positive predictor of movement-evoked pain at 6 months (R(2) = 0.04; St. β = 0.231, P = .004), despite no age group differences in outcome. Older age was found to be the strongest predictor of 3- and 6-month movement-evoked pain. Older adults may experience more pain related to movement, as well as endogenous pain excitation, in the first few months after shoulder arthroscopy. Future age-related research should consider use of movement-evoked pain intensity and experimental pain response as pain outcomes, as well as the utility of such measures in clinical care.


Vertebral augmentation in patients with multiple myeloma: a pooled analysis of published case series.
Studies examining the efficacy of vertebroplasty and kyphoplasty in patients with vertebral fractures from multiple myeloma are limited. We sought to perform a systematic review of published case studies examining changes in pain, disability, and analgesic drug use in patients with multiple myeloma who have undergone vertebral augmentation. We performed a pooled analysis of published case series of vertebral augmentation in patients with multiple myeloma. Twenty-three studies (9 kyphoplasty, 12 vertebroplasty, and 2 of both) with data on 923 patients were identified from a PubMed search. Quantitative outcome data included the Visual Analog Scale, the Brief Pain Inventory, the Short Form 36 Health Survey, and the Owestry Disability Index. Time periods were consolidated into 3: postoperatively ≤1 week, 1 week to 1 year, and ≥1 year. Change in analgesic use was also studied. Data were compared by using nonparametric tests and matched t tests for temporally linked data. Patients achieved a decrease in pain across all consolidated time periods. Pain, as measured on a 10-point scale, decreased by 4.8 points up to 1 week, 4.6 points up to 1 year, and 4.4 points after a year (P < .001). Decrease in pain was apparent early after treatment and was sustained with time. Kyphoplasty and vertebroplasty were equally effective in reducing pain scores because differences between procedures for each time period were insignificant (P < .9 for <1 week, P < 1.0 for ≤1 year, and P < .9 for >1 year. Our analysis demonstrates that vertebral augmentation is effective in patients with multiple myeloma.


Evaluation of liver and spleen stiffness of healthy dogs by use of two-dimensional shear wave elastography.
To assess liver and spleen stiffness in healthy dogs by use of a novel 2-D shear wave elastography (SWE) technique and to investigate the repeatability and reproducibility of the technique. 8 healthy adult Beagles. 2-D SWE was performed on each dog to assess liver and spleen stiffness. Repeatability (intraday variability) and reproducibility (interday variability) of 2-D SWE were investigated. For all 8 dogs, 2-D SWE was performed 3 times in 1 day (4-hour intervals) and on 3 separate days (1-week interval). Data were expressed as mean ± SD values for shear wave velocity and the Young modulus in the liver and spleen. Intraday and interday coefficients of variation were assessed for all variables. Mean ± SD shear wave velocity obtained for the liver and spleen was 1.51 ± 0.08 m/s and 2.18 ± 0.27 m/s, respectively. Mean value for the Young modulus obtained for the liver and spleen was 6.93 ± 0.79 kPa and 14.66 ± 3.79 kPa, respectively. Elasticity values were significantly higher for the spleen than for the liver. Intraday and interday coefficients of variation for all variables were < 25% (range, 3.90% to 20.70%). 2-D SWE was a feasible technique for assessing liver and spleen stiffness of healthy dogs. Future studies on the application of 2-D SWE for dogs with chronic hepatitis, cirrhosis, and portal hypertension are needed to evaluate the clinical applicability of 2-D SWE.


Migration of Beryllium via Multiple Exposure Pathways among Work Processes in Four Different Facilities.
Inhalation of beryllium is associated with the development of sensitization; however, dermal exposure may also be important. The primary aim of this study was to elucidate relationships among exposure pathways in four different manufacturing and finishing facilities. Secondary aims were to identify jobs with increased levels of beryllium in air, on skin, and on surfaces; identify potential discrepancies in exposure pathways, and determine if these are related to jobs with previously identified risk. Beryllium was measured in air, on cotton gloves, and on work surfaces. Summary statistics were calculated and correlations among all three measurement types were examined at the facility and job level. Exposure ranking strategies were used to identify jobs with higher exposures. The highest air, glove, and surface measurements were observed in beryllium metal production and beryllium oxide ceramics manufacturing jobs that involved hot processes and handling powders. Two finishing and distribution facilities that handle solid alloy products had lower exposures than the primary production facilities, and there were differences observed among jobs. For all facilities combined, strong correlations were found between air-surface (rp ≥ 0.77), glove-surface (rp ≥ 0.76), and air-glove measurements (rp ≥ 0.69). In jobs where higher risk of beryllium sensitization or disease has been reported, exposure levels for all three measurement types were higher than in jobs with lower risk, though they were not the highest. Some jobs with low air concentrations had higher levels of beryllium on glove and surface wipe samples, suggesting a need to further evaluate the causes of the discrepant levels. Although such correlations provide insight on where beryllium is located throughout the workplace, they cannot identify the direction of the pathways between air, surface, or skin. Ranking strategies helped to identify jobs with the highest combined air, glove, and/or surface exposures. All previously identified high-risk jobs had high air concentrations, dermal mass loading, or both, and none had low dermal and air. We have found that both pathways are relevant. [Supplementary materials are available for this article. Go to the publisher's online edition of Journal of Occupational and Environmental Hygiene for the following free supplemental resource: a file describing the forms of beryllium materials encountered during production and characteristics of the aerosols by process areas.].


Downregulation of CDK5 Restores Sevoflurane-Induced Cognitive Dysfunction by Promoting SIRT1-Mediated Autophagy.
An increasing number of studies have found that use of traditional anesthetics may lead to cognitive impairment of the immature brain. Our previous studies verified that cyclin-dependent kinase 5 (CDK5) plays a role in sevoflurane-induced cognitive dysfunction. Autophagy was shown to protect against anesthesia-induced nerve injury. Therefore, the current study aimed to ascertain if autophagy participates in anesthesia-induced neurotoxicity. In this study, primary hippocampal neurons were isolated and utilized for experiments in vitro. We also performed in vivo experiments with 6-day-old wild-type mice treated with or without roscovitine (Rosc, a CDK5 inhibitor) or 3-methyladenine (3-Ma, an autophagy inhibitor) after exposure to sevoflurane. We used the Morris water maze to analyze cognitive function. Immunohistochemical staining was used to assess pathologic changes in the hippocampus. The results showed that suppressing CDK5 reversed sevoflurane-induced nerve cell apoptosis both in vivo and in vitro and demonstrated that inhibits CDK5 activation promoted Sirtuin 1 (Sirt1) expression, which functions importantly in induced autophagy activation. Suppression of Sirt1 expression inhibited the protective effect of Rosc on sevoflurane-induced nerve injury by inhibiting autophagy activation. Our in vivo experiments also found that pretreatment with 3-Ma attenuated the protective effect of Rosc on sevoflurane-induced nerve injury and cognitive dysfunction. We conclude that inhibits CDK5 activation restored sevoflurane-induced cognitive dysfunction by promoting Sirt1-mediated autophagy.


Low-dose Aspirin for Primary Prevention of Cardiovascular Events in Postmenopausal Women with Type-2 Diabetes: The Prescriptive Approach in the Real World.
The long-term efficacy of low-dose aspirin for primary prevention of cardiovascular (CV) events in postmenopausal women with type-2 diabetes is controversial. Therefore, it is recommended only on an individual basis, recommendation of grade C. We enrolled 275 consecutive postmenopausal women with type-2 diabetes, without an increased bleeding risk and without preexisting CV disease as coronary artery disease, stroke, and peripheral vascular disease, but with a high risk assessed by score >10%, aged 60-69 years. All were receiving aspirin (75-100 mg daily), aspirin group (AG). 170 postmenopausal women with type-2 diabetes and without preexisting cardiovascular (CV) disease, but not on aspirin treatment, despite a high risk assessed by score >10%, were control group (CG). Mean age was 66 ± 4 years for AG and 65 ± 7 years for CG. Our goal was to identify the prevalence of low-dose aspirin prescriptions in these populations according to different clinical conditions. Women with only high risk were 41/275 (15%) on AG and 72/170 (42.3%) on CG, Chi-squared 41, Odds ratio 0.2, c.i. 95%, P < 0.0001. Women affected by metabolic syndrome were 105/275 (38.1%) on AG and 47/170 (27.6%) on CG, Chi-squared 5.1, Odds ratio 1.6, c.i. 95%, P < 0.02. Women affected by metabolic cardiomyopathy were 111/275 (40.3%) on AG and 44/170 (25.9%) on CG, Chi-squared 8, Odds ratio 1.8, c.i. 95%, P < 0.004. Women affected by diabetic cardiomyopathy were 18/275 (6.6%) on AG and 7/170 (4.2%) on CG, Chi-squared 1.2, Odds ratio 16, c.i. 95%, P < 0.2 n.s. Low-dose aspirin in our population is prescribed preferentially in postmenopausal women with type-2 diabetes when affected by metabolic syndrome or metabolic cardiomyopathy, at the opposite women with only high risk have lower chance to receive aspirin.


Evaluation of covered metallic stents in malignant biliary stenosis--prominent effectiveness in gallbladder carcinoma.
The survival time of patients with unresectable malignant biliary stenosis and the patent period of metallic biliary stents are different in each disease. The efficacy of the covered metallic stent was analyzed according to the primary disease. Seventy-three patients with bile duct carcinoma (12 cases), gallbladder carcinoma (22 cases), and pancreas carcinoma (39 cases) were retrospectively enrolled. Covered metallic stents were used in 42 patients and uncovered metallic stents in 31 patients. The patency of covered stents was compared with that of uncovered stents for each disease. The patent rate at 6 months after insertion was 80.6% (95% CI [72.6%, 88.6%]) for the covered stent, and 49.5% (95% CI [37.6%, 61.4%]) for the uncovered stent. The mean patent periods of the covered stent and the uncovered stent were 14.6 and 27.6 months for bile duct carcinoma (p=0.424), 12.7 and 3.0 months for gallbladder carcinoma (p=0.003), and 11.9 and 9.6 months for pancreas carcinoma (p=0.919), respectively. The covered metallic stent was the most effective in patients with gallbladder carcinoma.


Mistaken assumptions and missed opportunities: correlates of undiagnosed HIV infection among black and Latino men who have sex with men.
To identify demographic, behavioral, and psychological variables associated with being HIV positive unaware among black and Latino men who have sex with men (MSM). Participants recruited in 3 cities completed a computer-assisted interview and were tested for HIV infection (OraSure Technologies, Bethlehem, PA). HIV-positive unaware MSM were compared with MSM who tested HIV negative in bivariate and multivariate analyses. Of 1208 MSM (597 black and 611 Latino), 11% were HIV-positive unaware (18% black; 5% Latino). In multivariate analysis of the Latino MSM, being HIV-positive unaware was associated with nongay identity, high perceived risk of currently being HIV positive, and belief that sex with other Latino men reduces HIV transmission risk. Among black MSM, being HIV-positive unaware was associated with gay identity, moderately higher income, having health insurance, sexuality disclosure to a current health care provider, fewer than 3 lifetime HIV tests, high perceived risk of testing HIV positive, and belief that sex with other black men reduces HIV transmission risk. HIV prevention efforts should address misperceptions among those black and Latino MSM who believe that assortative (ie, intraracial) sexual mixing reduces risk of HIV infection. Our findings also revealed missed opportunities to diagnose black MSM with HIV infection who were already engaged in care and had disclosed their sexuality to their health care provider. Clinicians should offer HIV testing to all MSM, particularly black MSM, who disclose engaging in recent sex with other men to facilitate earlier diagnosis of HIV infection and reduce transmission risk to sexual partners.


Low birth weight in Spain associated with sociodemographic factors.
To describe the effect of different social and demographic characteristics on low birth weight (LBW) (less than 2500 g) in Spain, in both preterm (less than 37 weeks' gestation) and term infants (between 37 and 42 weeks' gestation). The study used data obtained from the Spanish birth registry. The study was based on those live born infants registered in 1988 from provinces where the birthweight details were completed in at least 99.5% of the birth registration records. A total of 1332 preterm LBW infants, 1292 term LBW infants, and 38,967 controls were included in the study. The odds ratio (OR) calculated by logistic regression was used as the measure of association between LBW and the sociodemographic variables. The highest ORs of preterm LBW were found in mothers younger than 20 years (1.32; 95% CI 0.98, 1.77) and older than 34 years (1.28; 95% CI 1.04, 1.59), in unmarried mothers (1.68; 95% CI 1.36, 2.07), and in fathers with manual occupations (1.26; 95% CI 1.08, 1.46). In term, live born infants the highest ORs were found in adolescent mothers (1.63; 95% CI 1.25, 2.14), in first born live born infants (1.38; 95% CI 1.09, 1.74) or the fourth born or more (1.28; 95% CI 0.91, 1.80), in unmarried mothers (1.55; 95% CI 1.27, 1.90), in housewives (1.13; 95% CI 0.99, 1.29), and in fathers with manual occupations (1.21; 95% CI 1.04, 1.42). The results have allowed documentation of the risk of preterm and term LBW in various age and social groups in Spain.


The intricate roles of RCC1 in normal cells and cancer cells.
RCC1 (regulator of chromosome condensation 1) is a highly conserved chromatin-binding protein and the only known guanine-nucleotide exchange factor of Ran (a nuclear Ras homolog). RCC1 plays an essential role in the regulation of cell cycle-related activities such as nuclear envelope formation, nuclear pore complex and spindle assembly, and nucleocytoplasmic transport. Over the last decade, increasing evidence has emerged highlighting the potential relevance of RCC1 to carcinogenesis, especially cervical, lung, and breast cancer. In this review, we briefly discuss the roles of RCC1 in both normal and tumor cells based on articles published in recent years, followed by a brief overview of future perspectives in the field.


Co-delivery of natural metabolic inhibitors in a self-microemulsifying drug delivery system for improved oral bioavailability of curcumin.
In spite of its well-documented anticancer chemopreventive and therapeutic activity, the clinical development of curcumin has been limited by its poor oral bioavailability. Curcumin has low aqueous solubility and undergoes extensive first pass metabolism following oral dosing. We hypothesized that oral bioavailability of curcumin can be enhanced by increasing its absorption and decreasing its metabolic clearance simultaneously. To test this hypothesis, we formulated curcumin with naturally occurring UGT inhibitors (piperine, quercetin, tangeretin, and silibinin) in a self-microemulsifying drug delivery system (SMEDDS). Mouse liver microsome studies showed that silibinin and quercetin inhibited curcumin glucuronidation effectively. When dosed orally in mice, the SMEDDS containing curcumin alone increased curcumin glucuronide concentrations in plasma without significantly affecting parent drug concentration. Of the four inhibitors examined in vivo, silibinin significantly improved the Cmax (0.15 μM vs. 0.03 μM for curcumin SMEDDS) and the overall bioavailability (3.5-fold vs. curcumin SMEDDS) of curcumin. Previous studies have shown that silibinin has anticancer activity as well. Thus, co-delivery of silibinin with curcumin in SMEDDS represents a novel and promising approach to improve curcumin bioavailability.


Cervical intraepithelial neoplasia is associated with genital tract mucosal inflammation.
Clinical studies demonstrate increased prevalence of human papillomavirus (HPV)-associated disease in HIV-infected individuals and an increased risk of HIV acquisition in HPV-infected individuals. The mechanisms underlying this synergy are not defined. We hypothesize that women with cervical intraepithelial neoplasia (CIN) will exhibit changes in soluble mucosal immunity that may promote HPV persistence and facilitate HIV infection. The concentrations of immune mediators and endogenous anti-Escherichia coli activity in genital tract secretions collected by cervicovaginal lavage were compared in HIV-negative women with high-risk HPV-positive (HRHPV+) CIN-3 (n = 37), HRHPV+ CIN-1 (n = 12), or PAP-negative control subjects (n = 57). Compared with control subjects, women with CIN-3 or CIN-1 displayed significantly higher levels of proinflammatory cytokines including interleukin (IL)-1α, IL-1β, and IL-8 (P < 0.002) and significantly lower levels of anti-inflammatory mediators and antimicrobial peptides, including IL-1 receptor antagonist, secretory leukocyte protease inhibitor (P < 0.01), and human β defensins 2 and 3 (P < 0.02). There was no significant difference in endogenous anti-E. coli activity after controlling for age and sample storage time. HRHPV+ CIN is characterized by changes in soluble mucosal immunity that could contribute to HPV persistence. The observed mucosal inflammation suggests a mechanism that may also contribute to the epidemiologic link between persistent HPV and HIV.