The application of hierarchical cluster analysis to the selection of isomorphous crystals.
It is generally assumed that the quality of X-ray diffraction data can be improved by merging data sets from several crystals. However, this effect is only valid if the data sets used are from crystals that are structurally identical. It is found that frozen macromolecular crystals very often have relatively low structure identity (and are therefore not isomorphous); thus, to obtain a real gain from multi-crystal data sets one needs to make an appropriate selection of structurally similar crystals. The application of hierarchical cluster analysis, based on the matrix of the correlation coefficient between scaled intensities, is proposed for the identification of isomorphous data sets. Multi-crystal single-wavelength anomalous dispersion data sets from four different protein molecules have been probed to test the applicability of this method. The use of hierarchical cluster analysis permitted the selection of batches of data sets which when merged together significantly improved the crystallographic indicators of the merged data and allowed solution of the structure.


Analysis of the complete nucleotide sequence of the picornavirus Theiler's murine encephalomyelitis virus indicates that it is closely related to cardioviruses.
Theiler's murine encephalomyelitis viruses (TMEV) are naturally occurring enteric pathogens of mice which constitute a separate serological group within the picornavirus family. Persistent TMEV infection in mice provides a relevant experimental animal model for the human demyelinating disease multiple sclerosis. To provide information about the TMEV classification, genome organization, and protein processing map, we determined the complete nucleotide sequence of the TMEV genome and deduced the amino acid sequence of the polyprotein coding region. The RNA genome, which is typical of the picornavirus family, is 8,098 nucleotides long. The 5' untranslated region is 1,064 nucleotides long (making it the longest in the picornavirus family after the aphthoviruses) and lacks a poly(C) tract. Computer-generated comparison of the 5' and 3' noncoding regions and polyprotein revealed the highest level of nucleotide and predicted amino acid identity between the TMEV and the cardioviruses encephalomyocarditis virus (EMCV) and Mengo virus. The TMEV polyprotein, which appears to be processed like EMCV since the amino acids flanking the putative proteolytic cleavage sites have been conserved, begins with a short leader peptide followed by 11 other gene products in the standard L-4-3-4 picornavirus arrangement. Because of these similarities, we propose that the TMEV be grouped with the cardioviruses. However, since TMEV and EMCV have different biophysical properties and show no cross-neutralization, they most likely belong in a separate cardiovirus subgroup.


Assistive Techniques for Proximal Anastomosis in Minimally Invasive Coronary Artery Bypass Grafting.
We introduce assistive techniques for proximal anastomosis in off-pump minimally invasive coronary artery bypass grafting (MICS CABG) to overcome difficult access to the ascending aorta in MICS CABG. An 8-cm left thoracotomy is made in the fifth intercostal space. ThoraTrak retractor (Medtronic Inc, Minneapolis, MN USA) is used to open the thoracotomy and is pulled to the cephalad and rightward direction toward to the ascending aorta. The pericardium is opened from the ascending aorta to the left ventricular apex and to the inferior vena cava. Two retraction sutures on the pericardial edge are used to laterally displace the heart. After dissecting between the ascending aorta and main pulmonary artery, the Octopus tissue stabilizer (Medtronic Inc, Minneapolis, MN USA), of which the suction tip is bent 60 degrees, is used to retract the pulmonary artery caudally. A flexible side-biting clamp (Vitalitec Inc.) is placed on the ascending aorta, and proximal anastomoses are handsewn on the ascending aorta. A total of 31 proximal anastomoses were completed with this technique between November 2013 and June 2015. All proximal anastomosis was completed without any difficulty. In MICS CABG, the technical challenges in proximal anastomosis due to difficult access to the aorta can be overcome safely by using this technique.


A randomized controlled trial comparing a modified Burch procedure and a suburethral sling: long-term follow-up.
The aim of this study was to compare the long-term results of a modified Burch procedure with a sling procedure for the treatment of stress incontinence with a low-pressure urethra. Thirty-six women with urodynamic stress incontinence, low-pressure urethra, urethral hypermobility and no significant pelvic organ prolapse were randomly assigned to undergo either a modified Burch procedure ( n=19) or a sling placement ( n=17). Cure of the stress incontinence (defined as a negative stress test and negative pad-weight test) was the primary long-term endpoint. Secondary endpoints included subjective cure of stress incontinence (defined as no incontinence episodes on a 1-week voiding diary) and voiding function studies. Comparisons of group means were performed with the Mann-Whitney U-test, pooled variance t-tests and separate variance t-tests. Proportions were compared with Fisher's exact test. A logistic regression analysis was performed to control for covariates that differed in our two groups despite randomization. Long-term follow-up (mean=72.6 months) was available for 82% (28/34) of the original study group. The objective cure rates for the Burch and sling groups were 84.6% and 100%, respectively ( P=0.17). Mean uroflowmetry rates for the Burch and sling groups were 7.38 and 6.8 ml/s, respectively ( P=0.58, 95%CI -2.5, 4.4). Mean postvoid residual volumes for both groups were 35 ml ( P=0.97, 95% CI -23.8, 65.9). Two sling patients (12%) required partial resection of their slings because of erosion. Both patients remained continent. In terms of voiding function and stress incontinence cure, there were no differences between groups undergoing modified Burch or sling procedures for treatment of urodynamic stress incontinence with low-pressure urethra.


Upper thermal limits of insects are not the result of insufficient oxygen delivery.
Most natural environments experience fluctuating temperatures that acutely affect an organism's physiology and ultimately a species' biogeographic distribution. Here we examine whether oxygen delivery to tissues becomes limiting as body temperature increases and eventually causes death at upper lethal temperatures. Because of the limited direct, experimental evidence supporting this possibility in terrestrial arthropods, we explored the effect of ambient oxygen availability on the thermotolerance of insects representing six species (Acheta domesticus, Hippodamia convergens, Gromphadorhina portentosa, Pogonomyrmex occidentalis, Tenebrio molitor, and Zophobus morio), four taxonomic orders (Blattodea, Coleoptera, Hymenoptera, and Orthoptera), and multiple life stages (e.g., adults vs. larvae or nymphs). The survival curves of insects exposed to temperatures (45° or 50°C) under normoxic conditions (21% O(2)) were compared with those measured under altered oxygen levels (0%, 10%, 35%, and 95% O(2)). Kaplan-Meier log rank analyses followed by Holm-Sidak pairwise comparisons revealed that (1) anoxia sharply diminished survival times in all groups studied, (2) thermotolerance under moderate hyperoxia (35% O(2)) or moderate hypoxia (10% O(2)) was the same as or lower than that under normoxia, (3) half of the experimental treatments involving extreme hyperoxia (95% O(2)) caused reduced thermotolerance, and (4) thermotolerance differed with developmental stage. Adult G. portentosa exhibited much higher thermotolerance than their first-instar nymphs, but responses from larval and adult Z. morio were equivocal. We conclude that some factor(s) separate from oxygen delivery is responsible for death of insects at upper lethal temperatures.


Introduction of Methyl Group in Substituted Isoselenazolones: Catalytic and Mechanistic Study.
Copper-catalyzed direct selenation of substituted 2-bromo-N-phenylbenzamide substrates with elemental selenium powder provided a series of methoxy-substituted isoselenazolones via the C-Se and Se-N bond formations. Phenolic substituted isoselenazolones have been obtained by O-demethylation of the corresponding methoxy-substituted analogues using boron tribromide. Some isoselenazolones have been structurally characterized by X-ray single-crystal analysis. The glutathione peroxidase (GPx)-like antioxidant activity of isoselenazolones has been evaluated both in thiophenol and coupled-reductase assays. All isoselenazolones showed good GPx-like activities in the coupled-reductase assay. The ferric-reducing antioxidant power of phenolic antioxidants has also been evaluated. The best phenolic antioxidants were found to be good ferric-reducing antioxidant power agents. The single electron transfer, hydrogen atom transfer, and proton-coupled electron transfer mechanisms for the antioxidant properties of all catalysts have been supported by density functional theory calculations. The catalytic cycle was proposed for one of the phenolic isoselenazolones involving diselenide, selenenyl sulfide, selenol, and selenenic acid as intermediates using 77Se{1H} NMR spectroscopy.


An extended model of vesicle fusion at the plasma membrane to estimate protein lateral diffusion from TIRF microscopy images.
Characterizing membrane dynamics is a key issue to understand cell exchanges with the extra-cellular medium. Total internal reflection fluorescence microscopy (TIRFM) is well suited to focus on the late steps of exocytosis at the plasma membrane. However, it is still a challenging task to quantify (lateral) diffusion and estimate local dynamics of proteins. A new model was introduced to represent the behavior of cargo transmembrane proteins during the vesicle fusion to the plasma membrane at the end of the exocytosis process. Two biophysical parameters, the diffusion coefficient and the release rate parameter, are automatically estimated from TIRFM image sequences, to account for both the lateral diffusion of molecules at the membrane and the continuous release of the proteins from the vesicle to the plasma membrane. Quantitative evaluation on 300 realistic computer-generated image sequences demonstrated the efficiency and accuracy of the method. The application of our method on 16 real TIRFM image sequences additionally revealed differences in the dynamic behavior of Transferrin Receptor (TfR) and Langerin proteins. An automated method has been designed to simultaneously estimate the diffusion coefficient and the release rate for each individual vesicle fusion event at the plasma membrane in TIRFM image sequences. It can be exploited for further deciphering cell membrane dynamics.


Disordered, stretched, and semiflexible biopolymers in two dimensions.
We study the effects of intrinsic sequence-dependent curvature for a two-dimensional semiflexible biopolymer with short-range correlation in intrinsic curvatures. We show exactly that when not subjected to any external force, such a system is equivalent to a system with a well-defined intrinsic curvature and a proper renormalized persistence length. We find the exact expression for the distribution function of the equivalent system. However, we show that such an equivalent system does not always exist for the polymer subjected to an external force. We find that under an external force, the effect of sequence disorder depends upon the averaging order, the degree of disorder, and the experimental conditions, such as the boundary conditions. Furthermore, a short to moderate length biopolymer may be much softer or has a smaller apparent persistent length than what would be expected from the "equivalent system." Moreover, under a strong stretching force and for a long biopolymer, the sequence disorder is immaterial for elasticity. Finally, the effect of sequence disorder may depend upon the quantity considered.


The Application and Development of Mural Art in Urban Public Environment Landscape Design.
With the gradual improvement of social and economic levels, cities not only meet people's basic living needs, but also have an artistic expression to a certain extent, and the public pays more attention to the beauty of the external environment. Landscapes in cities are settlement landscapes characterized by man-made landscapes, which are not only a cultural resource but also an ecological environment resource. Murals are also the first works of art of mankind. According to records, murals are the largest number of artworks in China. It is a popular work of art, representing social functions with historical value, aesthetic value, political significance, and educational function. Due to the great influence of the latest technology and emerging technology on urban art concepts, murals are widely used in urban public landscapes in Europe and America. Its grand and strong space coverage, not wasting too much space, and its imperceptible influence on vision make it the main element of urban public art. Therefore, the mural design in the urban public environment can improve China's current urban atmosphere and enhance urban characteristics. Bring the citizens a historical and cultural understanding of the urban environment they live in and enjoy art. According to the interrelationship between the city and the public environment and the human mural art, based on the relevant theoretical knowledge of urban planning and design, urban public art management, and based on the basic characteristics of human mural art, this study combines a large number of domestic and foreign cases, starting from the characteristics of the natural environment and human urban environment. This article studies the relationship between mural design and architectural design concepts and concepts in urban public environment landscape design.


Effects of Ocimum tenuiflorum on Induced Testicular Degeneration by Filgrastim in Wistar Rats.
Since the creation of man on earth, herbal remedies have been used as an invaluable, safe, and available natural source of medicine for the treatment of several dysfunctions in living animals and human beings. The pharmacological properties of these herbs are commonly known to include analgesic, antidiabetic, antispasmodic, hepatoprotective, reproductive, and cardioprotective potentials. Various nutritional, environmental, and physiological factors can affect the male reproductive system. Several herbal remedies are reported to target the testis at the spermatogenesis and hormonal level. Most of the chemotherapy drugs used for treating cancers cause adverse effects on male reproductive functions. Filgrastim is used for the treatment of cancer in patients suffering from neutropenia. Ocimum tenuiflorum is one of the herbal remedies used as a natural antioxidant substance for protecting the body organs against the toxic effects of chemotherapy drugs. Therefore, the current study was designed to investigate the possible productive effects of the O. tenuiflorum against adverse effects of filgrastim on testicular tissues in male Wistar rats. In total, 40 adult male rats were selected and randomly divided into four groups (n=10). Group 1 was treated with intraperitoneal administration of filgrastim at a dose of 30.83 µg/kg/day for 1 week. Group 2 received O. tenuiflorum suspended in corn oil and administered by gavages at 20 mg/kg/day for 1 week. Group 3 was intraperitoneally injected with filgrastim at a dose of 30.83 µg/kg/day for 1 week and treated with O. tenuiflorum at similar manure to group 2. Finally, group 4 was treated with placebo (0.9% saline solution). The results showed that filgrastim administration leads to the degeneration of spermatozoa and germ cells in the testicles of rats. The results of the current study showed that O. tenuiflorum has some ameliorating effects on the testicles and fertility of the rats which were treated with filgrastim. Finally, the recorded data showed that O. tenuiflorum has protective effects on testis tissues and reproductive functions in male rats.


Composition of explosives by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry.
Commercial explosives are complex mixtures that contain not only the active explosive agent(s) but also a host of other organic and inorganic compounds. The ultrahigh mass resolving power (m/delta m50% >200,000) and mass accuracy (<1 ppm) of electrospray ionization Fourier transform ion cyclotron resonance (ESI FTICR) mass spectrometry allow for definitive identification of various species in TNT, RDX, and HMX. We are thereby able to correct prior misassignments of the elemental compositions of the most abundant negative ions from electrospray of RDX and HMX. Although the (known) active agents of many explosives may be identified by low-resolution MS or MS/MS, it is the other characteristic components (indigenous or artificial additives) whose presence and elemental composition can potentially identify the source of the product. ESI FTICR mass spectrometry of smokeless powder, TNT, and Powermite resolves and identifies numerous nonactive ingredients, many of which are recovered in a postblast residue. In contrast, the residue recovered from an explosion of military C4 yielded several species derived from RDX but virtually none from other ingredients.


Tissue Inhibitor of Metalloproteinase-2 Suppresses Collagen Synthesis in Cultured Keloid Fibroblasts.
Keloids are defined as a kind of dermal fibroproliferative disorder resulting from the accumulation of collagen. In the remodeling of extracellular matrix, the balance between matrix metalloproteinases (MMPs) and the tissue inhibitors of metalloproteinases (TIMPs) is as critical as the proper production of extracellular matrix. We investigate the role of TIMPs and MMPs in the pathogenesis of keloids and examine the therapeutic potential of TIMP-2. The expression of TIMPs and MMPs in most inflamed parts of cultured keloid fibroblasts (KFs) and peripheral normal skin fibroblasts (PNFs) in the same individuals and the reactivity of KFs to cyclic mechanical stretch were analyzed by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay (n = 7). To evaluate the effect of treating KFs with TIMP-2, collagen synthesis was investigated by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay, and microscopic analysis was used to examine the treatment effects of TIMP-2 on ex vivo cultures of keloid tissue (n = 6). TIMP-2 was downregulated in cultured KFs compared with PNFs in the same individuals, and the reduction in TIMP-2 was exacerbated by cyclic mechanical stretch. Administration of TIMP-2 (200 or 300 ng/mL) significantly suppressed expression of Col1A2 and Col3A1 mRNA and collagen type I protein in KFs. TIMP-2 also significantly reduced the skin dermal and collagen bundle thickness in ex vivo cultures of keloid tissue. These results indicated that downregulation of TIMP-2 in KFs is a crucial event in the pathogenesis of keloids, and the TIMP-2 would be a promising candidate for the treatment of keloids.


Host Specificity of Flagellins from Segmented Filamentous Bacteria Affects Their Patterns of Interaction with Mouse Ileal Mucosal Proteins.
Segmented filamentous bacteria (SFB) are known modulators of the mammalian immune system. Currently, the technology for investigating SFB culture in vitro is immature, and as a result, the mechanisms of SFB colonization and immune regulation are not yet fully elucidated. In this study, we investigated the gene diversity and host specificity of SFB flagellin genes. The fliC1 and fliC2 genes are relatively conserved, while the fliC3 and fliC4 genes are more variable, especially at the central and C-terminal regions. Host specificity analysis demonstrated that the fliC1 genes do not cluster together based on the host organism, whereas the fliC3 and fliC4 genes were host specific at the nucleotide and deduced amino acid levels. SFB flagellin protein expression in the ileum mucosa and cecal contents was detected by using fluorescence in situ hybridization (FISH) combined with immunohistochemical (IHC) analysis, immunoblotting, and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Although the purified SFB FliC3 protein originating from both mouse and rat was able to activate Toll-like receptor 5 (TLR5)-linked NF-κB signaling, no host specificity was observed. Interestingly, the patterns of interaction with mouse ileum mucosal proteins were different for mouse FliC3 (mFliC3) and rat FliC3 (rFliC3). Gene Ontology (GO) and KEGG analyses indicated that more adherence-related proteins interacted with mFliC3, while more lysosome- and proteolysis-related proteins interacted with rFliC3. In vitro degradation experiments indicated that the stability of rFliC3 was lower than that of mFliC3 when they were incubated with mouse ileum mucosal proteins. In summary, the gene diversity and host specificity of SFB flagellin genes were investigated, and SFB flagellin expression was detected in gut samples.IMPORTANCE Since SFB genomes contain only one copy of each FliC gene, the diversity of FliC is representative of SFB strain diversity. Currently, little is known regarding the diversity and specificity of members of the group of SFB. The work presented herein demonstrates that select SFB strains, exhibiting unique FliC patterns, are present in a variety of mammalian hosts. SFB fliC genes were found to interact with a number of unique targets, providing further evidence for SFB host selection. Together, this work represents a major advancement in identifying SFB and delineating how members of the group of SFB interact with the host. Future examination of FliC genes will likely enhance our knowledge of intestinal colonization by the gut microbiota.


Hemozoin induces lung inflammation and correlates with malaria-associated acute respiratory distress syndrome.
Malaria-associated acute respiratory distress syndrome (MA-ARDS) is a deadly complication of malaria, and its pathophysiology is insufficiently understood. Both in humans and in murine models, MA-ARDS is characterized by marked pulmonary inflammation. We investigated the role of hemozoin in MA-ARDS in C57Bl/6 mice infected with Plasmodium berghei NK65, P. berghei ANKA, and P. chabaudi AS. By quantifying hemozoin in the lungs and measuring the disease parameters of MA-ARDS, we demonstrated a highly significant correlation between pulmonary hemozoin concentrations, lung weights, and alveolar edema. Histological analysis of the lungs demonstrated that hemozoin is localized in phagocytes and infected erythrocytes, and only occasionally in granulocytes. Species-specific differences in hemozoin production, as measured among individual schizonts, were associated with variations in pulmonary pathogenicity. Furthermore, both pulmonary hemozoin and lung pathology were correlated with the number of infiltrating inflammatory cells, an increased pulmonary expression of cytokines, chemokines, and enzymes, and concentrations of alveolar vascular endothelial growth factor. The causal relationship between hemozoin and inflammation was investigated by injecting P. falciparum-derived hemozoin intravenously into malaria-free mice. Hemozoin potently induced the pulmonary expression of proinflammatory chemokines (interferon-γ inducible protein-10/CXC-chemokine ligand (CXCL)10, monocyte chemotactic protein-1/CC-chemokine ligand 2, and keratinocyte-derived chemokine/CXCL1), cytokines (IL-1β, IL-6, IL-10, TNF, and transforming growth factor-β), and other inflammatory mediators (inducible nitric oxide synthase, heme oxygenase-1, nicotinamide adenine dinucleotide phosphate- oxidase-2, and intercellular adhesion molecule-1). Thus, hemozoin correlates with MA-ARDS and induces pulmonary inflammation.


Non-invasive diagnosis of liver steatosis using controlled attenuation parameter (CAP) and transient elastography.
Recently, a study showed that Controlled Attenuation Parameter (CAP), evaluated with transient elastography, could efficiently separate steatosis grades. The aim of this study was to prospectively evaluate the performance of CAP for the diagnosis of steatosis in patients with chronic liver disease. Consecutive patients with chronic liver disease had steatosis diagnosis using CAP, blood sample and liver biopsy. Steatosis was graded as the percentage of hepatocytes with fat: S0 ≤ 10%, S1: 11 ~ 33%, S2: 34 ~ 66%, S3 ≥ 67%. Characteristics of the 112 patients included were as follows: age 54 years, BMI 26 kg m(-) ², HCV 36%, NAFLD 25%. Steatosis repartition was: S0 52%, S1 19%, S2 14%, S3 15%. CAP was significantly correlated with SteatoTest, Fatty Liver Index (FLI), percentage of steatosis on liver biopsy, steatosis grade and slightly with liver stiffness, but not with fibrosis and activity grade on liver biopsy. Using CAP vs SteatoTest vs FLI score, Area Under the Receiver-Operating Characteristics (ROC) curves (AUROC)s were 0.84 vs 0.72 vs 0.72 for the diagnosis of steatosis ≥ S1, 0.86 vs 0.73 vs 0.71 for the diagnosis of steatosis ≥ S2, and 0.93 vs 0.73 vs 0.75 for the diagnosis of steatosis S3 respectively. For a sensitivity ≥ 90%, cut-offs of CAP were 215 dB m(-1) for S ≥ 1, 252 dB m(-1) for S ≥ 2, and 296 dB m(-1) for S3. CAP is very efficient to detect even low grade steatosis. CAP being implemented on FibroScan(®) (Echosens, Paris, France), both steatosis and fibrosis can be evaluated simultaneously, enlarging the spectrum of non-invasive techniques for the management of chronic liver diseases.


Livedoid vasculopathy - current aspects of diagnosis and treatment of cutaneous infarction.
Livedoid vasculopathy is a rare, chronic, recurrent disease of the cutaneous microcirculation. Its typical clinical manifestation is a triad which consists of livedo racemosa of the skin, episodic painful ulcerations of the distal aspects of the legs and a healing process leaving small porcelain-white scars called atrophie blanche. As an important result of recent research, livedoid vasculopathy has been defined as a coagulation disorder classified as a vasculopathy different from inflammatory vasculitis. This differentiation adds to the current pathophysiologic understanding and supports the therapeutic rationale with respect to the use of new systemic anticoagulants. The prevention of irreversible residual scarring and the improvement of patientsí quality of life are the main goals in treating cutaneous infarction and require early and consequent treatment. This article presents current knowledge on diagnosing this rare disease and offers practical guidance on its therapy.


[Diagnostic problems in a patient with staphylococcal endocarditis].
A patient, aged 68, admitted to the Internal Medicine Ward, in connection with syncope and a persisting fever which had appeared for the third time during the last two months. The patient had an artificial aortic valve graft implanted and a heart bypass surgery (CAPG) in 1994. A urine examination, performed before his admission to hospital, showed the characteristic changes which appeared in UTI (urine tract infection). Then the patient was treated with quinolone (zanocin) and the abatement of the fever was achieved. After two weeks the recurrence of the intermittent fever appeared. The diagnosis of bacterial endocarditis was established and a broad spectrum antibiotic treatment was introduced and continued in spite of the negative blood culture. The circulatory failure and growing cachexia developed in the patient and led to his death within two weeks. From the last blood sample Staphylococcus aureus was cultured. In autopsy the phlegmonic and haemorrhagic changes were found, with foci of necrosis in the region of the implanted artificial valve graft. Some changes of the type of interstitial numerous, microfocal, phlegmonic foci were also found. The author drew the conclusion that if the treatment with broad spectrum antibiotics had been introduced, in the patient with a fever, after a heart surgery, there would have been a chance for the successful treatment.


Prevalence of Burnout Among Pain Medicine Physicians and Its Potential Effect upon Clinical Outcomes in Patients with Oncologic Pain or Chronic Pain of Nononcologic Origin.
To evaluate the prevalence of burnout among physicians treating patients with chronic pain and to assess the potential relationships between the presence of burnout and patients' clinical outcomes such as pain relief, satisfaction with pain control, and quality of life. An observational, prospective, and noncomparative study. Pain medicine clinics. Physicians from medical departments involved in the management of chronic pain. Patients aged ≥18 years who exhibited moderate chronic pain lasting at least three months. Physicians were evaluated with the Maslach Burnout Inventory-Human Services Survey (MBI-HSS). Patients were evaluated with the Charlson Comorbidity Index, the Brief Pain Inventory-Short Form (BPI-SF), the EuroQol-5D (EQ-5D), and ad hoc instruments for evaluating satisfaction with pain control, the extent to which the treatment met patients' expectations, and subjective impressions of improvement. Of the 301 physician participants, 22 (7.3%, 95% confidence interval [CI] = 4.9 to 10.8) met the criteria of burnout. Burnout was higher among physicians from pain units, while none of the 35 primary care physicians reported burnout. The presence of burnout was positively associated with patients' pain relief (odds ratio [OR] = 1.423, 95% CI = 1.090 to 1.858) but not with satisfaction with pain control or quality of life. Of the remaining independent variables, being treated by pain unit physicians was significantly associated with worse pain relief (OR = 0.592, 95% CI = 0.507 to 0.691), lower satisfaction (β = -0.680, 95% CI = -0.834 to -0.525), and worse quality of life (β = -4.047, 95% CI = -5.509 to -2.585) compared with being treated by physicians from other specialties (e.g., traumatologists, oncologists, etc.). Our study shows a lack of negative or clinically relevant (as shown by the negligible to small effect sizes) impact of burnout on patient-reported outcomes (namely, pain relief, satisfaction, and quality of life) in patients with chronic pain who are treated by pain medicine physicians.


Immunocytochemical localization of three vesicular glutamate transporters in the cat retina.
Vesicular transporters play an essential role in the packaging of glutamate for synaptic release and so are of particular importance in the retina, where glutamate serves as the neurotransmitter for photoreceptors, bipolar cells, and ganglion cells. In the present study, we have examined the distribution of the three known isoforms of vesicular glutamate transporter (VGLUT) in the cat retina. VGLUT1 was localized to all photoreceptor and bipolar cells, whereas VGLUT2 was found in ganglion cells. This basic pattern of complementary distribution for the two transporters among known populations of glutamatergic cells is similar to previous findings in the brain and spinal cord. However, the axon terminals of S-cone photoreceptors were found to express both VGLUT1 and VGLUT2 and some ganglion cells labeled for both VGLUT2 and VGLUT3. Such colocalizations suggest the existence of dual modes of regulation of vesicular glutamate transport in these neurons. Staining for VGLUT2 was also present in a small number of varicose processes, which were seen to ramify throughout the inner plexiform layer. These fibers may represent axon collaterals of ganglion cells. The most prominent site of VGLUT3 immunoreactivity was in a population of amacrine cells; the axon terminals of B-type horizontal cells were also labeled at their contacts with rod spherules. The presence of the VGLUT3 transporter at sites not otherwise implicated in glutamate release may indicate novel modes of glutamate signaling or additional roles for the transporter molecule.


Nurse prescribing in diabetes.
The adjustment of diabetes medication is vital for the prevention of complications. Insulin dose adjustment by diabetes specialist nurses is common practice but its legality is questionable. Two surveys were undertaken to identify common practice. The results were used to establish the basis for an effective multiprofessional protocol for nurse administration of diabetes medication.


[Analysis of demands of African students for China-Africa malaria prevention training].
To understand the demands of African students on the China-Africa malaria prevention training programs as well as explore further suggestions on the student selection and course content design. A self-administered questionnaire survey was conducted, and all the students who attended in the malaria prevention training courses in 2014 and 2015 were included. The Chi-square test was conducted to analyse the correlations between professional backgrounds, work statues and training needs. A total of 161 individuals were sampled eventually. These participants were trained in either English (58.4%) or French (41.6%). Most of the participants were male (69.3%), the major of them were mainly clinical technology specialty (40.0%), and most of them worked in malaria area within 10 years (56.4%). Moreover, 48.2% of the participants used more than 76% of total work time on malaria control, and more than 80% worked in national or provincial/municipal level. The working areas of these participants were focused on clinical field (41.4%) and official field (29.9%), and only a few of them were from research positions (11.9%). The most needed course content in malaria training was strategy and epidemiology knowledge for malaria prevention and control (65.5%), while clinical workers were most needed to be trained (39.2%). The participants who came from French speaking countries preferred strategy training (χ2 = 12.528, P < 0.01), and those worked in the national level were aslo more likely to choose strategies training course (χ2 = 10.508, P < 0.05). Currently, the China-Africa malaria prevention training programs could basically satisfy African students'needs. However, more aimed courses should be designed according to their professional backgrounds, national situation, work experiences on malaria control, and institutional levels.


Annulated dinuclear metal-free and Zn(II) phthalocyanines: photophysical studies and quantum mechanical calculations.
The results of steady-state and time-resolved absorption and fluorescence experiments as well as quantum mechanical density functional theory (DFT) calculations of metal-free and Zn(II) mononuclear and dinuclear (sharing a common benzene ring) phthalocyanines are presented. A detailed comparison between measured and calculated absorption spectra of all compounds is done, showing a good agreement between theory and experiment. The NH tautomerization for phthalocyanines with an extended pi-electron system was shown for the first time at room temperature. The photophysical properties of all possible NH tautomers of metal-free dinuclear Pc have been fully characterized. In the first tautomer, Pc(parallel), both pairs of hydrogen atoms are parallel to the connection line of two Pc units. The maximum of the lowest-energy Q absorption band, lambda abs, in Pc(parallel) is located at 832 nm, whereas the spectral position of the fluorescence maximum lies at lambdafl=837 nm. The second NH tautomer, Pc(perpendicular) (lambdaabs=853 nm, lambdafl=860 nm), presents the two pairs of hydrogen atoms perpendicularly orientated to the covalent axis, and the third one, Pc(mix) (lambdaabs=864 nm, lambdafl=872 nm), contributing in a minor extend to the absorption and fluorescence spectra of the metal-free dinuclear phthalocyanine, has one perpendicular and one parallel pair of hydrogen atoms. Obviously, only one configuration exists in the case of the Zn(II)-containing dinuclear phthalocyanine (lambdaabs=845 nm, lambdafl=852 nm).


Amphiregulin-EGFR signaling regulates PTHrP gene expression in breast cancer cells.
Parathyroid hormone-related protein (PTHrP) is an autocrine/paracrine factor produced by breast cancer cells that is speculated to play a major role in permitting breast cancer cells to grow into the bone microenvironment by stimulating the bone resorption axis. It has been previously shown that EGFR signaling induces the production of PTHrP in several primary and transformed epithelial cell types. Therefore, we investigated the relationship between EGFR and PTHrP gene expression in human breast cancer cells. Of a panel of 7 breast epithelial and cancer cell lines, the osteolytic, EGFR- positive lines (MDA-MB-231 and NS2T2A1) exhibited higher levels of PTHrP transcript expression. Amphiregulin mRNA levels in all lines were approximately 2 orders of magnitude higher than those of TGFalpha or HB-EGF. In the EGFR bearing lines, the receptor was phosphorylated at tyrosine 992 under basal conditions, and the addition of 100 nM amphiregulin did not lead to the phosphorylation of other tyrosine residues typically phosphorylated by the prototypical ligand EGF. Treatment of the EGFR positive lines with the EGFR inhibitor PD153035 and amphiregulin-neutralizing antibodies reduced PTHrP mRNA levels by 50-70%. Stable EGFR expression in the MCF7 line failed to increase basal PTHrP mRNA levels; however, treatment of this cell line with exogenous EGF or amphiregulin increased PTHrP transcription 3-fold. Transient transfection analysis suggests that the MAPK pathway and ETS transcription factors mediate EGFR coupling to PTHrP gene expression. Taken together, it appears that autocrine stimulation of EGFR signaling by amphiregulin is coupled to PTHrP gene expression via EGFR Tyr992 and MAPK, and that this pathway may contribute to PTHrP expression by breast tumor cells.


[Human in vitro fertilisation and embryotransfer: first results at the 2nd department of obstetrics and gynaecology, university of vienna (author's transl)].
Methods and Results of human in vitro fertilisation at the 2nd Department of Obstetrics and Gynaecology, University of Vienna are reported. Applying preparations of media and culture techniques according to Edwards et al. no satisfactory results could be achieved: only 3 out of 11 preovulatory oocytes were fertilised, no cleavages occurred. Using modified Ham's F 10 medium according to Lopata et al. fertilisation could be observed in 60% as well as regular cleavages in 40%. 5 Embryotransfers have been performed in the 8 cell or in the 16 cell stage respectively. No implantation occurred in 4 of these, in one case evidence for a transient implantation could be proved by an increased activity of SP1 and HCG beta. In this case obviously postimplantation death occurred resulting in a bleeding 16 days after oocyte recovery. Possible reasons for failures in fertilisation and cleavage as well as possibilities for improving the rate of reimplantation are discussed.


[PECULIARITY OF NATIONAL TUBERCULOSIS PROGRAM, JAPAN--Public-Private Mix from the Very Beginning, and Provision of X-ray Apparatus in Most General Practitioner's Clinics].
Modern National Tuberculosis Program (NTP) of Japan started in 1951 when Tuberculosis (TB) Control Law was legislated, and 3 major components were health examination by tuberculin skin test (TST) and miniature X-ray, BCG vaccination and extensive use of modern TB treatment. As to the treatment program, Japan introduced Public-Private Mix (PPM) from the very beginning, and major reasons why PPM was adopted are (1) TB was then highly prevalent (Table 1), (2) TB sanatoria where many specialists are working are located in remote inconvenient places due to stigma against TB, (3) health centers (HCs) in Japan are working exclusively on prophylactic activities, and minor exceptions are treatment of sexually transmitted diseases and artificial pneumothorax for TB cases, however, as it covers on the average 100,000 population, access is not so easy in rural area, (4) Out-patients clinics mainly operated by general practitioners (GPs) are located throughout Japan, and the access is easy. Methods of TB treatment was developing rapidly in early 1950s, however, in 1952, as shown in Table 2, artificial pneumothorax and peritoneum were still used in many cases, and to fix the dosage of refill air, fluoroscopy was needed. Hence, GPs treating TB under TB Control Law had to be equipped with X-ray apparatus. To maintain the quality of TB treatment, "Criteria for TB treatment" was provided and revised taking into consideration the progress in TB treatment. If applied methods of treatment fit with the above criteria, public support is made for the cost of TB treatment. To discuss the applied treatment, TB Advisory Committee was set in each HC, composing of 5 members, director of HC, 2 TB specialists and 2 doctors recommended by the local medical association. In 1953, the first TB prevalence survey using stratified random sampling method was carried out, and the prevalence of TB requiring treatment was estimated at 3.4%, and only 21% of found cases knew their own disease, and more than half of all TB were found above 30 years of age. Based on these results, mass screening was expanded to cover whole population in 1955, and since 1957, cost of mass screening and BCG vaccination was covered 100% by public fund. Unified TB registration system covering whole Japan was introduced in 1961, and in the same year, national government subsidy for the hospitalization of infectious TB cases was raised from 50% to 80%. Hence, Japan succeeded to organize PPM system in TB care, and with 10% annual decline of TB, in 1975, Japan moved into the TB middle prevalence country.


Nerve compression injuries due to traumatic false aneurysm.
Experience with 17 patients with delayed onset of compression neuropraxia due to hemorrhage following nonoperative treatment of penetrating arterial injuries is presented. Fifteen cases involved the arteries of the neck shoulder girdle and upper extremity and two the gluteal vessels. This resulted in dysfunction of components of the brachial plexus, median ulnar, and sciatic nerves. Follow-up extended from 3 to 18 months. Of 10 brachial plexus lesions two recovered fully, five partially, and three not at all. Of seven peripheral nerve injuries, full recovery occurred in two patients and none in five. Adverse prognostic factors for neurological recovery are sepsis, involvement of intrinsic hand innervation and the sciatic nerve. An improved prognosis may be expected for upper trunk lesions of the brachial plexus and radial nerve lesions. The complication is essentially avoidable and a careful appraisal of the circulatory status must be made in all patients with penetrating trauma in the neck and shoulder girdle and buttock.


Heterogeneity, Characteristics, and Public Health Implications of Listeria monocytogenes in Ready-to-Eat Foods and Pasteurized Milk in China.
Listeria monocytogenes is a foodborne pathogen with a high mortality rate in humans. This study aimed to identify the pathogenic potential of L. monocytogenes isolated from ready-to-eat (RTE) foods and pasteurized milk in China on the basis of its phenotypic and genotypic characteristics. Approximately 7.7% (44/570) samples tested positive for L. monocytogenes among 10.8% (39/360) RTE and 2.4% (5/210) pasteurized milk samples, of which 77.3% (34/44) had < 10 MPN/g, 18.2% (8/44) had 10-110 MPN/g, and 4.5% (2/44) had > 110 MPN/g. A total of 48 strains (43 from RTE foods and five from milk samples) of L. monocytogenes were isolated from 44 positive samples. PCR-serogroup analysis revealed that the most prevalent serogroup was II.2 (1/2b-3b-7), accounting for 52.1% (25/48) of the total, followed by serogroup I.1 (1/2a-3a) accounting for 33.3% (16/48), serogroup I.2 (1/2c-3c) accounting for 12.5% (6/48), and serogroup II.1 (4b-4d-4e) accounting for 2.1%. All isolates were grouped into 11 sequence types (STs) belonging to 10 clonal complexes (CCs) and one singleton (ST619) via multi-locus sequence typing. The most prevalent ST was ST87 (29.2%), followed by ST8 (22.9%), and ST9 (12.5%). Virulence genes determination showed that all isolates harbored eight virulence genes belonging to Listeria pathogenicity islands 1 (LIPI-1) (prfA, actA, hly, mpl, plcA, plcB, and iap) and inlB. Approximately 85.4% isolates carried full-length inlA, whereas seven isolates had premature stop codons in inlA, six of which belonged to ST9 and one to ST5. Furthermore, LLS (encoded by llsX gene, representing LIPI-3) displays bactericidal activity and modifies the host microbiota during infection. LIPI-4 enhances neural and placental tropisms of L. monocytogenes. Results showed that six (12.5%) isolates harbored the llsX gene, and they belonged to ST1/CC1, ST3/CC3, and ST619. Approximately 31.3% (15/48) isolates (belonging to ST87/CC87 and ST619) harbored ptsA (representing LIPI-4), indicating the potential risk of this pathogen. Antimicrobial susceptibility tests revealed that > 95% isolates were susceptible to 16 antimicrobials; however, 60.4 and 22.9% isolates were intermediately resistant to streptomycin and ciprofloxacin, respectively. The results show that several isolates harbor LIPI-3 and LIPI-4 genes, which may be a possible transmission route for Listeria infections in consumers.


Thermodynamic behavior and structural properties of an aqueous sodium chloride solution upon supercooling.
We present the results of a molecular dynamics simulation study of thermodynamic and structural properties upon supercooling of a low concentration sodium chloride solution in TIP4P water and the comparison with the corresponding bulk quantities. We study the isotherms and the isochores for both the aqueous solution and bulk water. The comparison of the phase diagrams shows that thermodynamic properties of the solution are not merely shifted with respect to the bulk. Moreover, from the analysis of the thermodynamic curves, both the spinodal line and the temperatures of maximum density curve can be calculated. The spinodal line appears not to be influenced by the presence of ions at the chosen concentration, while the temperatures of maximum density curve displays both a mild shift in temperature and a shape modification with respect to bulk. Signatures of the presence of a liquid-liquid critical point are found in the aqueous solution. By analyzing the water-ion radial distribution functions of the aqueous solution, we observe that upon changing density, structural modifications appear close to the spinodal. For low temperatures, additional modifications appear also for densities close to that corresponding to a low density configurational energy minimum.


Photocurable 3D-printed PMBG/TCP biphasic scaffold mimicking vasculature for bone regeneration.
Mesoporous bioglass (MBG) with excellent osteointegration, osteoinduction, and biodegradability is a promising material for bone regeneration. However, its clinical application is hindered by complex processing and a lack of personalization, low mechanical strength, and uncontrollable degradation rate. In this study, we developed a double-bond-functionalized photocurable mesoporous bioglass (PMBG) sol that enabled ultrafast photopolymerization within 5 s. By further integrating nanosized tricalcium phosphate (TCP) particles through three-dimensional (3D) printing technology, we fabricated personalized and highly porous PMBG/TCP biphasic scaffolds. The mechanical properties and degradation behavior of the scaffolds were regulated by varying the amount of TCP doping. In vitro and in vivo experiments verified that PMBG/TCP scaffolds slowly released SiO44- and Ca2+, forming a vascularized bone regeneration microenvironment within the fully interconnected pore channels of the scaffold. This microenvironment promoted angiogenesis and accelerated bone tissue regeneration. Overall, this work demonstrates the solution to the problem of complex processing and lack of personalization in bioglass scaffolds, and the developed PMBG/TCP biphasic scaffold is an ideal material for bone regeneration applications with broad clinical prospects.


Connecting perceived economic threat and prosocial tendencies: The explanatory role of empathic concern.
Recent research suggests that perceived economic threat constitutes a valid predictor of people's attitudes and behaviors. While accumulated empirical evidence has mostly underlined the deleterious psychological effects (e.g., reduced psychological well-being) of perceived economic threat in times of economic strain, we postulate that individuals experiencing higher economic threat linked to the Spanish economic crisis are more prone to engage in other-beneficial prosocial behavior. Across two independently collected community samples, we tested this theoretical formulation and examined the potential mediating roles of empathic concern (Studies 1 & 2) and identification (Study 2). Study 1 (N = 306) revealed that participants who descended in the social scale due to the negative national economic context were engaged in a larger number of helping behaviors over the last three months compared to participants who did not descend the social ladder-independently of several sociodemographic and ideological factors. Moreover, our data indicated these effects were driven by increased empathic concern. Study 2 (N = 588), in which two hypothetical helping-behavior scenarios were randomly administered (crisis-related vs. control), showed that participants under high perceived financial threat exhibited an undifferentiated pattern of prosociality. However, moderated-mediation analyses indicated that empathic concern explained the perceived financial threat-helping behavior link in the hypothetical crisis-related scenario but not in the hypothetical control scenario. Together, these findings extend prior literature on the psychosocial effects of perceived economic threat and the determinants of other-oriented behavior. Implications of these findings and suggestions for further research are discussed.


Drug resistance by evasion of antiangiogenic targeting of VEGF signaling in late-stage pancreatic islet tumors.
Function-blocking antibodies to VEGF receptors R1 and R2 were used to probe their roles in controlling angiogenesis in a mouse model of pancreatic islet carcinogenesis. Inhibition of VEGFR2 but not VEGFR1 markedly disrupted angiogenic switching, persistent angiogenesis, and initial tumor growth. In late-stage tumors, phenotypic resistance to VEGFR2 blockade emerged, as tumors regrew during treatment after an initial period of growth suppression. This resistance to VEGF blockade involves reactivation of tumor angiogenesis, independent of VEGF and associated with hypoxia-mediated induction of other proangiogenic factors, including members of the FGF family. These other proangiogenic signals are functionally implicated in the revascularization and regrowth of tumors in the evasion phase, as FGF blockade impairs progression in the face of VEGF inhibition.


The vertebrate lysozyme inhibitor Ivy functions to inhibit the activity of lytic transglycosylase.
The proteinaceous inhibitor of vertebrate lysozymes (Ivy) is produced by a collection of Gram-negative bacteria as a stress response to damage to their essential cell wall component peptidoglycan. A paralog of Ivy, Ivyp2 is produced exclusively by a number of pseudomonads, including Pseudomonas aeruginosa, but this protein does not inhibit the lysozymes, and its function was unknown. In this study, we demonstrate that the production of Ivy (homologs of both Ivyp1 and Ivyp2) correlates with bacteria that do not O-acetylate their peptidoglycan, a modification that controls the activity of the lytic transglycosylases. Furthermore, we show that both Ivy proteins are potent inhibitors of the lytic transglycoslyases, enzymes involved in the biosynthesis and maintenance of peptidoglycan. These data suggest that the true physiological function of the Ivy proteins is to control the autolytic activity of lytic transglycosylases within the periplasm of Gram-negative bacteria that do not produce O-acetylated peptidoglycan and that the inhibition of exogenous lysozyme by Ivy is simply a fortuitous coincidence.


Acute epididymitis and Chlamydia trachomatis.
Prostatitis and epididymitis, which are the most common complications of urethritis in men below 35 years of age may be associated with Chlamydia trachomatis infection. Chlamydiae have been isolated from expressed prostatic fluid in men with urethritis complicated by prostatitis and epididymitis. Epididymitis seem to be an established complication of chlamydial urethritis particularly when urethritis is complicated with prostatitis. Since epididymitis may lead to reduced fertility, prompt antibiotic treatment using tetracyclines should be administered to patients with this condition.


Marrow transplantation following busulfan and cyclophosphamide in multiple myeloma.
We report the use of busulfan and cyclophosphamide as conditioning before allogeneic bone marrow transplantation for a 38-year-old man with multiple myeloma. Three months post-transplant the monoclonal IgA band previously present was no longer detectable in the serum and plasma cells were no longer visible in the marrow. The patient died on day 93 with fungal infection. We conclude that the combination of busulfan and cyclophosphamide could be valuable before transplant for other patients with multiple myeloma.


Immunoglobulin A protease activity of Ureaplasma urealyticum.
All of 14 serotype standards and 34 of 35 wild-type strains of Ureaplasma urealyticum isolated from humans demonstrated an immunoglobulin A (IgA) protease activity. This activity degraded radiolabeled human IgA including IgA1 but not IgG or azocasein. The IgA fragments were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by radioautography, and they had molecular weights of about 110,000 and 50,000. The IgA protease activity persisted in 25 mM EDTA but was sensitive to trypsin; it was presumed to be protein. This is the fourth microbial genus and the first myocoplasma species in which an IgA protease activity has been identified. Such activity was absent in Mycoplasma pneumoniae, Mycoplasma hominis, and Acholeplasma laidlawii.


Side rails as physical restraints: the need for appropriate assessment.
Side rails are commonly used in the care of the older person. They can be classified as a physical restraint if they restrict the movement of an individual and also if the individual has an inability to have them removed/lowered at will. Advanced age should not be an indicator for side rail use and an individualized approach in patient assessment is advised.


Genetic relationships between personality and eating attitudes and behaviors.
Genetic and environmental factors underlying relationships between personality traits and disordered eating were examined in 256 female adolescent twin pairs (166 monozygotic, 90 dizygotic). Eating behaviors were assessed with the Total Score, Body Dissatisfaction, Weight Preoccupation, Binge Eating, and Compensatory Behavior subscales from the Minnesota Eating Disorders Inventory (M-EDI; K. L. Klump, M. McGue, & W. G. Iacono, 2000). Personality characteristics were assessed with the Negative Emotionality, Positive Emotionality, and Constraint scales from the Multidimensional Personality Questionnaire (MPQ; A. Tellegen, 1982). Model-fitting analyses indicated that although genetic factors were more likely to contribute to MPQ and M-EDI phenotypic associations than environmental factors, shared genetic variance between the 2 phenotypes was limited. MPQ personality characteristics may represent only some of several genetic risk factors for eating pathology.


CRISPR-Cas systems exploit viral DNA injection to establish and maintain adaptive immunity.
Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems provide protection against viral and plasmid infection by capturing short DNA sequences from these invaders and integrating them into the CRISPR locus of the prokaryotic host. These sequences, known as spacers, are transcribed into short CRISPR RNA guides that specify the cleavage site of Cas nucleases in the genome of the invader. It is not known when spacer sequences are acquired during viral infection. Here, to investigate this, we tracked spacer acquisition in Staphylococcus aureus cells harbouring a type II CRISPR-Cas9 system after infection with the staphylococcal bacteriophage ϕ12. We found that new spacers were acquired immediately after infection preferentially from the cos site, the viral free DNA end that is first injected into the cell. Analysis of spacer acquisition after infection with mutant phages demonstrated that most spacers are acquired during DNA injection, but not during other stages of the viral cycle that produce free DNA ends, such as DNA replication or packaging. Finally, we showed that spacers acquired from early-injected genomic regions, which direct Cas9 cleavage of the viral DNA immediately after infection, provide better immunity than spacers acquired from late-injected regions. Our results reveal that CRISPR-Cas systems exploit the phage life cycle to generate a pattern of spacer acquisition that ensures a successful CRISPR immune response.


Inhibition of HtrA2/Omi ameliorates heart dysfunction following ischemia/reperfusion injury in rat heart in vivo.
High temperature requirement A2 (HtrA2)/Omi is a mitochondrial serine protease that is released into the cytosol from mitochondria and in turn promotes caspase activation by proteolyzing inhibitor of apoptosis proteins. Here we asked whether treatment with an HtrA2/Omi inhibitor, 5-[5-(2-nitrophenyl)furfuryliodine]-1,3-diphenyl-2-thiobarbituric acid (UCF-101), restores heart dysfunction following ischemia/reperfusion injury in vivo. Rats underwent a 30-min ischemia by occluding the left anterior descending artery, followed by 24 h reperfusion. UCF-101 (0.75 or 1.5 micromol/kg, i.p.) was administered 10 min before reperfusion. UCF-101 treatment significantly recovered the mean arterial blood pressure and ameliorated contractile dysfunction of the left ventricle 72 h after reperfusion with concomitant reduction of infarct size. Cardio-protection mediated by UCF-101 was correlated with reduced X-linked inhibitor of apoptosis protein (XIAP) degradation and inhibition of Caspase-9, Caspase-3, and Caspase-7 processing. Furthermore, UCF-101 prevented loss of membrane integrity by inhibiting fodrin breakdown in cardiomyocytes. UCF-101-induced cytoprotection was also correlated with reduced Fas ligand expression and inhibition of FLIP degradation following ischemia/reperfusion. These results suggest that UCF-101 rescues cardiomyocytes from ischemia/reperfusion injury by inhibiting XIAP degradation and Fas/Fas-ligand-induced apoptosis, thereby ameliorating ischemia/reperfusion-induced myocardial dysfunction.


Internal guide RNA interactions interfere with Cas9-mediated cleavage.
The CRISPR/Cas system uses guide RNAs (gRNAs) to direct sequence-specific DNA cleavage. Not every gRNA elicits cleavage and the mechanisms that govern gRNA activity have not been resolved. Low activity could result from either failure to form a functional Cas9-gRNA complex or inability to recognize targets in vivo. Here we show that both phenomena influence Cas9 activity by comparing mutagenesis rates in zebrafish embryos with in vitro cleavage assays. In vivo, our results suggest that genomic factors such as CTCF inhibit mutagenesis. Comparing near-identical gRNA sequences with different in vitro activities reveals that internal gRNA interactions reduce cleavage. Even though gRNAs containing these structures do not yield cleavage-competent complexes, they can compete with active gRNAs for binding to Cas9. These results reveal that both genomic context and internal gRNA interactions can interfere with Cas9-mediated cleavage and illuminate previously uncharacterized features of Cas9-gRNA complex formation.


First report of the efficacy of vestibular rehabilitation in improving function in patients with Neurofibromatosis type 2: an observational cohort study in a clinical setting.
Patients with Neurofibromatosis type 2 often experience debilitating neuro-otological problems which affect their mobility and balance. This study examined the efficacy of a personalised program of vestibular rehabilitation for patients with Neurofibromatosis type 2. An observational cohort study analysing routinely collected data for 21 patients in a highly specialised Neurofibromatosis type 2 service. Vestibular rehabilitation comprised an initial one-hour assessment followed by a patient-specific exercise program reviewed in person and by email consultations. Patients were subsequently followed-up at 9 months. The vestibular rehabilitation efficacy was assessed using the Dynamic Gait Index score. Nineteen of 21 patients were assessed as impaired and at risk of falls pre-rehabilitation (Dynamic Gait Index <19/24), of which 79% showed clinical improvement post-rehabilitation. There was a significant improvement in the Dynamic Gait Index scores pre-rehabilitation to post-rehabilitation (p < 0.001) and outcomes were subsequently maintained at the 9-month follow-up assessment. Whilst the pre-rehabilitation Dynamic Gait Index scores of patients with more severe genotype were lower compared to other patients, the beneficial effect of vestibular rehabilitation was similar amongst genetic severity groups. Personalised vestibular rehabilitation significantly improves function in Neurofibromatosis type 2, sustaining benefits for 9 months, irrespective of patients' age or genetic severity. Implications for rehabilitation Patients with Neurofibromatosis type 2 experience debilitating neuro-otological problems which affect their mobility and balance. A patient-tailored program of vestibular rehabilitation was offered in a highly specialised clinic for six months with a follow-up assessment at 9 months post-treatment. All patients improved from baseline, with 79% of them achieving clinically significant improvement in function and with statistically significant benefits sustained for 9 months. The beneficial effect of vestibular rehabilitation was similar for all patients, regardless of age or genetic severity, suggesting vestibular rehabilitation could be incorporated in routine clinical care in Neurofibromatosis type 2 clinics internationally.


Development of Non-Viral, Trophoblast-Specific Gene Delivery for Placental Therapy.
Low birth weight is associated with both short term problems and the fetal programming of adult onset diseases, including an increased risk of obesity, diabetes and cardiovascular disease. Placental insufficiency leading to intrauterine growth restriction (IUGR) contributes to the prevalence of diseases with developmental origins. Currently there are no therapies for IUGR or placental insufficiency. To address this and move towards development of an in utero therapy, we employ a nanostructure delivery system complexed with the IGF-1 gene to treat the placenta. IGF-1 is a growth factor critical to achieving appropriate placental and fetal growth. Delivery of genes to a model of human trophoblast and mouse placenta was achieved using a diblock copolymer (pHPMA-b-pDMAEMA) complexed to hIGF-1 plasmid DNA under the control of trophoblast-specific promoters (Cyp19a or PLAC1). Transfection efficiency of pEGFP-C1-containing nanocarriers in BeWo cells and non-trophoblast cells was visually assessed via fluorescence microscopy. In vivo transfection and functionality was assessed by direct placental-injection into a mouse model of IUGR. Complexes formed using pHPMA-b-pDMAEMA and CYP19a-923 or PLAC1-modified plasmids induce trophoblast-selective transgene expression in vitro, and placental injection of PLAC1-hIGF-1 produces measurable RNA expression and alleviates IUGR in our mouse model, consequently representing innovative building blocks towards human placental gene therapies.


Metabolic cooperation between mouse embryonal carcinoma cells and their differentiated derivatives.
Metabolic cooperation has been used as an index of intercellular communication between mouse embryonal carcinoma (EC) and other cell types. EC cells do not cooperate with differentiated cells of various properties or origins. In contrast they cooperate with cells of all other EC lines tested, including a human teratocarcinoma line, but with different efficiencies. This might reflect differences either in cell-cell interactions or in the formation of gap junctions. During in vitro differentiation, EC cells appear to remain isolated from differentiated cell types.


Fossil Bacteria in Pyrite.
A considerable variety of bacteria and similar microorganisms are present as preserved remains in pyrite samples of Pennsylvanian age obtained from the coal measures of southeastern Ohio. The types observed are similar to the present-day microflora found in stagnant pools of low oxygen and nutrient content.


Why study ageing in cold-blooded vertebrates?
Homeotherms exhibit programmed ageing correlated with their determinate mode of growth and the natural attrition of irreplaceable functional units in vitally essential organs. Poikilotherms are potentially capable of indefinite growth, can supplement their populations of functional units, and may therefore not be subject to the inherent depreciations of ageing. Comparative studies may enable us to determine which group is the exception to nature's rule.


Recommendations to protect patients and health care practices from Medicare and Medicaid fraud.
Fraud is defined as knowingly submitting, or causing to be submitted, false claims or making misrepresentations of a fact to obtain a federal health care payment for which no entitlement would otherwise exist. In today's health care environment, Medicare and Medicaid fraud is not uncommon. The negative impact of fraud is vast because it diverts resources meant to care for patients in need to the benefit of fraudsters. Fraud increases the overall costs for vital health care services and can potentially be harmful to Medicare and Medicaid beneficiaries. The objectives of this commentary are to describe the types and trends of Medicare and Medicaid fraud that are committed, and provide recommendations to protect patients and health care practices. Specifically, this article identifies types of Medicare and Medicaid fraud at beneficiary (patient) and provider level, and it can be intentional or unintentional. This article also describes the 3 primary laws that prohibit fraud and gives fraud case examples relevant to each law, including the False Claims Act, Anti-Kickback Statute, and the Stark Law. We also discuss currently trending and emerging areas, including opioid and pharmacogenetic testing; both have experienced heavier and higher-profile instances of fraud in today's health care landscape. Last, the article summarizes detection methods and recommendations for health care providers and patients to protect themselves against fraud. Recommended strategies to combat fraud are discussed at policy, practice, and grassroots levels. Health care practitioners, including pharmacists, can use these strategies to protect themselves and their patients from becoming victims of fraud or unknowingly committing fraud.


Patient participation in the intensive care unit.
Patient participation in healthcare is important for optimizing treatment outcomes and for ensuring satisfaction with care. The purpose of the study wasto explore critical care nurses' perceptions of patient participation for critically ill patients. Qualitative data were collected in four separate focus group interviews with 17 nurses from two hospitals. The interviews were analyzed using qualitative content analysis. Initially, the nurses stated that patient participation in the intensive care unit (ICU) was dependent on the patient's health condition and consciousness. However, during the interviews three descriptive categories emerged from their experience, that is: passive patient participation, one-way communication and nurse/patient interaction. In the ICU, the possibilities for patient participation in nursing care are not only dependent on the patient's health condition but also on the nurse's ability to include patients in various care actions despite physical and/or mental limitations. When the patient is not able to participate, nurses strive to achieve participation through relatives' knowledge and/or other external sources of information.


Comparison of medium- and high-energy collimators for 131I-tositumomab dosimetry.
Residence time measurements obtained by serial whole-body conjugate-view imaging are commonly used in patient-specific dosimetry for radioimmunotherapy applications. In order to determine the effect of collimator selection on residence time measurements for (131)I, the accuracies of (131)I half-life measurements obtained with multiple gamma-camera and collimator combinations were investigated. Serial anterior and posterior whole-body images were acquired over a period of 15 d with 4 different gamma-cameras and medium- or high-energy collimators. Background-corrected geometric mean counts from the images were fitted to a monoexponential curve to determine the half-life of (131)I obtained with the different gamma-camera and collimator combinations. An average half-life of 8.15 d (SD, 0.07 d) was obtained with all gamma-camera and collimator combinations. A half-life of 8.12 d (SD, 0.11 d) was obtained with the high-energy collimators, and a half-life of 8.18 d (SD, 0.04 d) was obtained with the medium-energy collimators. These values are all very close to the (131)I physical half-life of 8.02 d and were not found to be statistically significantly different (P = 0.44). Similar results were obtained for the half-life obtained with single-head gamma-camera configurations (mean half-life, 8.15 d; SD, 0.12 d). The therapeutic (131)I-tositumomab dose resulting from the differences in the measured half-life ranged from 2.58 to 2.6 GBq (69.8-70.4 mCi). There is no significant difference in (131)I half-life and residence time measurements obtained with medium- or high-energy collimators in dual-head or single-head imaging configurations.


A dramatic switch of enantioselectivity in asymmetric heck reaction by benzylic substituents of ligands.
A series of benzylic substituted P, N-ligands 1 and 2 have been synthesized. The Pd-complexes of these ligands show high catalytic activity and enantioselectivity in catalyzing the asymmetric Heck reaction. A dramatic switch in enantioselectivity is realized using ligands with and without substituents at the benzylic position of the ligand. Ligands 1 with H as the substituents offer products in (R)-configuration while ligands 2 with the methyl as substituents result in (S)-configuration products. In most cases high enantioselectivities are achieved. Density functional theory calculations on the reaction mechanism as well as X-ray analysis of 1a-PdCl 2 and 2a-PdCl 2 complexes provide a rational explanation for the above observations.


Clinical prognosis in BRAF-mutated PTC.
BRAF mutation has recently emerged as a potential prognostic marker for papillary thyroid carcinoma (PTC) due to several studies suggesting that it may condition the development of tumors with aggressive behavior. A study of the phenotypes of thyroid follicular cell lines and transgenic mice characterized by targeted expression of BRAF mutation indicates that, at variance with RET/PTC rearrangement, it induces or facilitates genomic instability and higher invasiveness and eventually deeper tumor de-differentiation and more significant suppression of apoptosis. An analysis of differential gene expression of PTCs harboring BRAF mutation versus PTCs characterized by other genetic alterations shows an important impairment of the expression of genes related to intra-thyroidal iodine metabolism machinery, up-regulation of Glut-1 mRNA, methylation-induced gene silencing of tumor suppressor genes and up-regulation of pro-angiogenetic proteins such as VEGF. Correlation of BRAF mutation with PTC clinico-pathological features yields controversial results, with several studies showing the association with unfavourable clinico-pathological qualities, while others do not confirm the findings. This review will summarize the studies in favor of or in contrast with a role of BRAF mutation as a prognostic marker in PTC. We will also indicate what information we still need in order to routinely introduce this indicator in clinical practice.


Localization of proteins to the apico-lateral junctions of Drosophila epithelia.
We have examined the distribution of proteins in the apico-lateral cell junctions in Drosophila imaginal discs. The subcellular distribution of these proteins in normal and mutant proliferating cells was analyzed with marker antibodies and confocal microscopy. Antibodies to phosphotyrosine (PY), Armadillo (Arm) and Drosophila E-cadherin (DE-cad) as well as FITC phalloidin marking filamentous actin, labeled the site of the adherens junction, whereas antibodies to Discs large (DIg), Fasciclin III (FasIII) and Coracle (Cor) labeled the more basal septate junction. The junctional proteins labeled by these antibodies underwent specific changes in distribution during the cell cycle. We have previously shown that a loss-of-function dlg mutation, which causes neoplastic imaginal disc overgrowth, leads to loss of the septate junctions and the formation of what appear to be ectopic adherens junctions [Woods et al., 1996]. We therefore extended this study to examine the effects of mutations in other genes that also cause imaginal disc overgrowth. Based on staining with PY and DIg antibodies, the apico-lateral junctional complexes appeared normal in tissue from the hyperplastic overgrowth mutants fat, dco, gd and wts. However, imaginal disc tissue from the neoplastic overgrowth mutants dlg and lgl showed abnormal distribution of the junctional markers including a complete loss of apico-basal polarity in loss-of-function dlg mutations. These results support the idea that some of the proteins of apico-lateral junctions are required both for apico-basal cell polarity and for the signalling mechanisms controlling cell proliferation, whereas others are required more specifically in cell-cell signalling.


Application of far-infrared irradiation in the manufacturing process of green tea.
Seven kinds of green tea leaves were manufactured with far-infrared (FIR) irradiation, and the physicochemical characteristics of the green tea were determined. Appropriate FIR irradiation during the manufacturing process significantly increased the polyphenolic content of green tea. FIR irradiation at 90 degrees C for 10 min, replacing the roasting step, and of the fully processed green tea leaves (GTP3) increased the total phenol content of green tea from 475.6 to 811.1 mg/g and the total flavanol content from 175.7 to 208.7 mg/g, as compared to the control. Epigallocatechin and epigallocatechin gallate increased from 57.68 and 9.60 mg/g in a nonirradiated control to 89.88 and 16.33 mg/g in GTP3, respectively. Ascorbic acid, caffeine, and nitrite scavenging activities were also increased in GTP3. However, the overall color change of GTP3 was negligible. These results indicate that the chemical properties of green tea are significantly affected by FIR irradiation at specific stages of the manufacturing process of green tea leaves and that this FIR irradiation results in high-quality green tea.


HIV-associated intussusception.
A 39-year-old man with untreated HIV presented with watery, non-bloody diarrhea and associated 25-lb weight loss. His workup was notable for a CT abdomen/pelvis which showed an entero-enteric intussusception of 3.4 cm without an underlying pathologic mass. The patient's intussusception resolved without intervention, as is typical of telescoping <3.5 cm. HIV is associated with an increased risk of developing intussusception because of an increased incidence of infectious and neoplastic conditions of the bowel.


Involuntary emotional expression disorder - new/old disease in psychiatry and neurology.
Involuntary emotional expression disorder (IEED) is underrecognized by clinicians, misdiagnosed as depression or bipolar disorder and undertreated, because clinicians are unfamiliar with the disorder. An important clinical consideration for IEED is that of distinguishing mood from affect. IEED describes a syndrome of relatively stereotypical episodes of uncontrollable crying and/or laughing, resulting from lesions of multiple types, in multiple brain regions, without an apparent stimulus to trigger such responses. This syndrome is common among a number of neurological diseases like patients with a stroke or traumatic brain injury (TBI), patients with amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), as well as dementias such as Alzheimer;s disease (AD), and motor disorders such as Parkinson;s disease (PD). The neuropathological cause and neurochemistry of the disorder remains unclear. There is general agreement that IEED is the result of an injury to the neurological pathways that control the expression of emotions. Adequate treatment can reduce the frequency and improve the quality of life of patients and caregivers.


Total Surgical Plication of Left Ventricular Aneurysm Using the BioVentrix Revivent Myocardial Anchoring System.
Surgical ventricular reconstruction (SVR) is the therapy of choice for patients with left ventricular dilatation, apical and anterolateral transmural scar, and low ejection fraction. STICH trial did not show that SVR led to improved survival but several observational studies did. However, because of the considerable operative risk, open heart surgery is considered risky in debilitated patients and clinical results are controversial. Alternative less invasive strategies for left ventricular aneurysm repair have been proposed. We present a case of a left ventricular aneurysm repair using the less invasive ventricular enhancement technique (LIVE) with the Revivent TC system (BioVentrix Inc., San Ramon, CA) in a totally surgical approach, instead of a hybrid interventional-surgical one, as previously described.


Effect of intensive statin therapy on regression of carotid intima-media thickness in patients with subclinical carotid atherosclerosis (a prospective, randomized trial: PEACE (Pitavastatin Evaluation of Atherosclerosis Regression by Intensive Cholesterol-lowering Therapy) study).
Atherosclerosis often advances before symptoms appear. It remains uncertain whether intensive cholesterol-lowering therapy with statin is beneficial when compared with moderate cholesterol-lowering therapy in patients with subclinical carotid atherosclerosis. The PEACE study was a prospective, randomized, open-labeled, blinded end points, two-arm parallel treatment group comparison study conducted at 15 centers in Japan. A total of 303 patients with carotid intima-media thickness (CIMT) thickening (>1.1 mm) whose low-density lipoprotein cholesterol (LDL-C) level was more than 100 mg/dl were enrolled, in which 223 patients completed the 12 months' follow-up study. Patients were randomly assigned to receive either moderate (target LDL-C; 100 mg/dl) or intensive (target LDL-C; 80 mg/dl) cholesterol-lowering therapy with pitavastatin. The primary end point was the change in mean far wall common CIMT. LDL-C level declined to 89.4 ± 20 mg/dl in the intensive group, while it declined to 95.1 ± 22.5 mg/dl in the moderate group at 12 months' follow-up (p < 0.05 between the groups). The change in mean CIMT was -0.024 (95% confidence interval -0.046 to -0.0014) mm/year (p < 0.05 vs. baseline) in the intensive group, and -0.0078 (95% confidence interval -0.028 to 0.012) mm/year (p = 0.4406 vs. baseline) in the moderate group. However, there was no significant difference in the change in mean far wall common CIMT between the groups (p = 0.29). Intensive cholesterol-lowering therapy did not show superior effects on the progression of CIMT to moderate cholesterol-lowering therapy, whereas only intensive cholesterol-lowering therapy regressed the carotid atherosclerosis over one year.


European nursing organizations stand up for family presence during cardiopulmonary resuscitation: a joint position statement.
Empirical evidence suggests that family presence during cardiopulmonary resuscitation (CPR) has beneficial effects. Although many American professional organizations have endorsed the idea of family presence, there is less formal support in Europe. In addition, the attitude of nurses from Anglo-Saxon countries, such as United Kingdom and Ireland, is more positive toward family presence than the attitude of nurses of mainland Europe. In order to support existing guidelines and to stimulate health care organizations to develop a formal policy with respect to family witnessed CPR, 3 important European nursing organizations have recently developed a joint position statement.


The clinical value of maternal and fetal Doppler parameters in low-risk postdates pregnancies - a prospective study.
The objective of this study is to determine the clinical significance of maternal and fetal ultrasound Doppler flow indices in postdates pregnancies. This prospective study comprised 120 low-risk pregnant women beyond 40 weeks of gestation. All participants underwent Doppler assessment including of fetal middle cerebral artery (MCA), umbilical, and uterine arteries. Perinatal outcomes were recorded and evaluated for possible associations with Doppler flow values. Adverse perinatal outcomes were defined as umbilical cord arterial pH <7.1, Apgar score <7 at 5 min, neonatal admission to a neonatal intensive care unit, and emergency cesarean section due to abnormal intrapartum cardiotocogram Results: Adverse perinatal outcome rate was 17.5% (n = 21). Doppler indices of umbilical artery, MCA, uterine artery and the cerebroplacental ratio (CPR) did not differ significantly between pregnant women with and without adverse perinatal outcomes. Neonatal birth weight was found to correlate negatively with umbilical artery Doppler indices, including the peak systolic to end diastolic ratio (p = .04), the resistance index (p = .02), and the pulsatility index (p = .01). Doppler values of the uterine artery, MCA and CPR did not correlate with neonatal birth weight. The contribution of maternal and fetal ultrasound Doppler to the prediction of adverse perinatal outcomes in low-risk postdates pregnancies is low. Hence, performing routine Doppler examination as part of postdates pregnancy assessments is unlikely to yield significant clinical benefit.


The Efficacy of Colchicine and Dapsone Combination Therapy in Relapsed Immune Thrombocytopenia.
The aim of the present paper is to evaluate the efficacy and safety of colchicine and dapsone combination therapy in cases of steroid-dependent, relapsed and refractory immune thrombocytopenia (ITP). This is a retrospective study of ITP patients who attended the Hematology Clinic at Chiang Mai University Hospital (Thailand) from 1 January 2008 to 30 September 2014. Medical records and clinical data were reviewed for efficacy and adverse effects. Sixty-four ITP patients received the combination therapy. The median age was 46 years and 70.3% were female. The majority (65.6%) were relapsed ITP patients. Median platelet count before starting treatment was 22.6×109/L. The response rate was 82.8%, with 75.0% of patients having a complete response. Median time to response was 8 weeks. The response rate was higher in relapsed patients (90.4%) compared to refractory (61.5%) and steroid-dependent patients (77.8%). Steroid treatment was discontinued in 30 patients (50%) following combination therapy. The most common side effect was hemolysis due to dapsone which was found in eight patients (12.5%). We can therefore conclude that combination therapy with colchicine and dapsone is an alternative second-line therapy option in relapsed ITP cases with acceptable side effects.


Genomic distribution of maize facultative heterochromatin marked by trimethylation of H3K27.
Trimethylation of histone H3 Lys-27 (H3K27me3) plays a critical role in regulating gene expression during plant and animal development. We characterized the genome-wide distribution of H3K27me3 in five developmentally distinct tissues in maize (Zea mays) plants of two genetic backgrounds, B73 and Mo17. There were more substantial differences in the genome-wide profile of H3K27me3 between different tissues than between the two genotypes. The tissue-specific patterns of H3K27me3 were often associated with differences in gene expression among the tissues and most of the imprinted genes that are expressed solely from the paternal allele in endosperm are targets of H3K27me3. A comparison of the H3K27me3 targets in rice (Oryza sativa), maize, and Arabidopsis thaliana provided evidence for conservation of the H3K27me3 targets among plant species. However, there was limited evidence for conserved targeting of H3K27me3 in the two maize subgenomes derived from whole-genome duplication, suggesting the potential for subfunctionalization of chromatin regulation of paralogs. Genomic profiling of H3K27me3 in loss-of-function mutant lines for Maize Enhancer of zeste-like2 (Mez2) and Mez3, two of the three putative H3K27me3 methyltransferases present in the maize genome, suggested partial redundancy of this gene family for maintaining H3K27me3 patterns. Only a portion of the targets of H3K27me3 required Mez2 and/or Mez3, and there was limited evidence for functional consequences of H3K27me3 at these targets.


The Vitamin D Metabolite Ratio Is Associated With Changes in Bone Density and Fracture Risk in Older Adults.
Recent studies have suggested that 25-hydroxyvitamin D (25(OH)D) may be a poor biomarker of bone health, in part because measured levels incorporate both protein-bound and free vitamin D. The ratio of its catabolic product (24,25-dihydroxyvitamin D [24,25(OH)2 D]) to 25(OH)D (the vitamin D metabolite ratio [VMR]) may provide more information on sufficient vitamin D stores and is not influenced by vitamin D-binding protein concentrations. We evaluated whether the VMR or 25(OH)D are more strongly associated with bone loss and fracture risk in older adults. We performed a retrospective cohort study of 786 community-dwelling adults aged 70 to 79 years who participated in the Health Aging and Body Composition study. Our primary outcomes were annual changes in bone density and incident fracture. The mean age of these participants was 75 ± 3 years, 49% were female, 42% were Black, and 23% had an estimated glomerular filtration rate (eGFR) <60 mL/mL/1.73m2 . In fully adjusted models, a 50% lower VMR was associated with 0.3% (0.2%, 0.6%) more rapid decline in total hip bone mineral density (BMD). We found similar relationships with thoracic and lumbar spine BMD. In contrast, 25(OH)D3 concentrations were not associated with longitudinal change in BMD. There were 178 fractures during a mean follow-up of 10 years. Each 50% lower VMR was associated with a 49% (95% confidence interval [CI] 1.06, 2.08) greater fracture risk, whereas lower 25(OH)D3 concentrations were not significantly associated with fracture risk (hazard ratio [HR] per 50% lower 1.07 [0.80, 1.43]). In conclusion, among a diverse cohort of community-dwelling older adults, a lower VMR was more strongly associated with both loss of BMD and fracture risk compared with 25(OH)D3 . Trials are needed to evaluate the VMR as a therapeutic target in persons at risk for worsening BMD and fracture. © 2021 American Society for Bone and Mineral Research (ASBMR).


Purification and characterization of a novel metalloproteinase, acurhagin, from Agkistrodon acutus venom.
Acurhagin, a high-molecular mass hemorrhagic metalloproteinase, was purified from the crude venom of Agkistrodon acutus using anion-exchange and hydrophobic interaction chromatography. Acurhagin is a monomer with a molecular mass of 51.4 kDa under non-reducing conditions on SDS-PAGE and 48,133 Da by mass spectrometry. Partial amino acid sequence of its metalloproteinase domain is homologous to other high-molecular mass metalloproteinases from snake venoms. It preferentially cleaved Aalpa chain of fibrinogen, followed by Bbeta chain, while gamma chains was minimally affected. Monitored by RP-HPLC, it extensively degraded fibrinogen into various peptide fragments. In aqueous solution, acurhagin autoproteolyzed to a 30 kDa fragment at 37 degrees C. The N-terminal sequence of the 30 kDa fragment of acurhagin showed a high homology to those proteins consisting of disintegrin-like and cysteine-rich domains. Caseinolytic assay showed that the proteinase activity of acurhagin was slightly enhanced by Ca2+ and Mg2+, but completely inhibited by Zn2+. When treated with metal chelators, acurhagin was completely inactivated. Furthermore, acurhagin exerts an inhibitory effect on ADP-induced platelet aggregation of platelet-rich plasma in an incubation-time dependent manner. It also impairs collagen- and ristocetin-induced platelet aggregation by cleaving collagen and vWF, respectively.


Self-selection and earnings assimilation: immigrants from the former Soviet Union in Israel and the United States.
Drawing on U.S. decennial census data and on Israeli census and longitudinal data, we compare the educational levels and earnings assimilation of Jewish immigrants from the former Soviet Union (FSU) in the United States and Israel during 1968-2000. Because the doors to both countries were practically open to FSU immigrants between 1968 and 1989, when FSU immigrants were entitled to refugee visas in the United States, the comparison can be viewed as a natural experiment in immigrants' destination choices. The results suggest that FSU immigrants to the United States are of significantly higher educational level and experience significantly faster rates of earnings assimilation in their new destination than their counterparts who immigrated to Israel. We present evidence that patterns of self-selection in immigration to Israel and the United States--on both measured and unmeasured productivity-related traits--is the main reason for these results. When the immigration regulations in the United States changed in 1989, and FSU Jewish immigrants to the United States had to rely on family reunification for obtaining immigrant visas, the adverse effects of the policy change on the type of FSU immigrants coming to the United States were minor and short-lived As early as 1992, the gaps in the educational levels between FSU immigrants coming to Israel and to the United States returned to their pre-1989 levels, and the differences in earnings assimilation of post-1989 immigrants in the United States and Israel are similar to the differences detected in the 1980s.


The Italian Registry of Laparoscopic Surgery of the Spleen (IRLSS) A retrospective review of 379 patients undergoing laparoscopic splenectomy.
In December 2000, the Italian Registry of Laparoscopic Surgery of the Spleen (IRLSS) was formally launched under the auspices of the Italian Society for Endoscopic Surgery and New Technologies (SICE). The aim of this multicentre study was to analyse various aspects of the treatment that are still under discussion, such as the extension of the laparoscopic indications in cases of malignancy, independently of the associated splenomegaly, patient selection and operative techniques. A retrospective review of 379 patients undergoing laparoscopic splenectomy for haematological diseases from February 1, 1993, to September 15, 2005, was conducted. Data were collected from the 18 italian centres participating in the IRLSS. The mean length of surgery was 140 minutes (range: 25-420). Conversion was necessary in 25 cases (6.6%), and at least one accessory spleen was found in 30 patients (8%). The mean spleen weight was 1200 g (range: 85-4500). Perioperative death occurred in two cases (0.5%). There were no complications in 312 patients (82.3%), with a mean hospital stay of 5.5 days (range: 2-30). Morbidity occurred in 67 patients (17.8%), mainly consisting in transient fever (n = 22), pleural effusions (n = 16), and actual or suspected haemorrhage (n = 14), requiring re-intervention in 7 patients. This first study carried out on the IRLSS data shows that laparoscopic splenectomy may constitute the gold standard for haematological diseases with a normal-sized spleen. The low morbidity and mortality rates suggest that laparoscopic splenectomy can be successfully proposed also for splenomegaly in haematological malignancies.


Immunogenicity and persistence of a targeted anti-caries DNA vaccine.
We have previously reported that a targeted anti-caries DNA vaccine, pGJA-P, induced accelerated and increased antibody responses compared with a non-targeted anti-caries DNA vaccine. Recently, pGJA-P/VAX, a new targeted anti-caries DNA vaccine for human trials, was constructed by replacing the pCI vector used in the construction of pGJA-P with pVAX1, the only vector authorized by the US Food and Drug Administration in clinical trials. Here, we report on our exploration of the kinetics of the antibody responses generated following pGJA-P/VAX immunization and the persistence of pGJA-P/VAX at both the inoculation site and the draining lymph nodes. Intranasal vaccination of mice with pGJA-P/VAX induced strong antibody responses that lasted for more than 6 months. Furthermore, pGJA-P/VAX could still be detected at both the inoculation site and the draining cervical lymph nodes 6 months after immunization. Thus, the persistent immune responses are likely due to the DNA depot in the host, which acts as a booster immunization.


Genetic differences in the ethanol sensitivity of GABAA receptors expressed in Xenopus oocytes.
Animal lines selected for differences in drug sensitivity can be used to help determine the molecular basis of drug action. Long-sleep (LS) and short-sleep (SS) mice differ markedly in their genetic sensitivity to ethanol. To investigate the molecular basis for this difference, mRNA from brains of LS and SS mice was expressed in Xenopus oocytes and the ethanol sensitivity of gamma-aminobutyric acid A (GABAA)- and N-methyl D-aspartate (NMDA)-activated ion channels was tested. Ethanol facilitated GABA responses in oocytes injected with mRNA from LS mice but antagonized responses in oocytes injected with mRNA from SS animals. Ethanol inhibited NMDA responses equally in the two lines. Thus, genes coding for the GABAA receptor or associated proteins may be critical determinants of individual differences in ethanol sensitivity.


Multinucleated giant cells in areas of the brain stem necrosis after cardiac arrest in two infants.
A rare type of cellular reaction in the brain stem of two infants with cardiac arrest encephalopathy is presented. After cardiac arrest both newborns were resuscitated and put on artificial ventilation. Their survival time amounted to 16 and 18 days, respectively, in a deep coma with areflexia. At the postmortem examination a widespread hemispheric necrosis of the gray and white matter was observed as well as symmetrical necrosis of the tegmental part of the brain stem extending from the midbrain up to medulla. Striking proliferation of blood vessels and large number of multinucleated giant cells originating from monocyte/macrophage lineage was found in the areas of the brain stem necrosis. No evidence of inflammatory process was found. It seems that giant cells appeared as local reaction on disintegration of maturing structures.


Child welfare caseworkers as brokers of mental health services: a pilot evaluation of Project Focus Colorado.
Youth in the child welfare system (CWS) have substantially higher rates of mental health needs compared to the general population, yet they rarely receive targeted, evidence-based practices (EBPs). Caseworkers play the critically important role of "service broker" for CWS youth and families. This study examines preliminary caseworker-level outcomes of Project Focus Colorado (PF-C), a training and consultation program designed to improve access to EBPs for CWS youth. PF-C evaluation occurred in four child welfare offices (two intervention [n = 16 caseworkers] vs. two practice-as-usual, wait-list control [WLC; n = 12 caseworkers]). Receipt of PF-C was associated with significantly increased caseworker knowledge of (a) EBPs, (b) child mental health problems, (c) evidence-based treatment components targeting mental health problem areas, and (d) mental health screening instruments, compared to WLC. Dose of training and consultation was associated with greater ability to correctly classify mental health problems and match them to EBPs. These preliminary results suggest that targeted training and consultation help to improve caseworker knowledge of children's mental health needs, EBPs for mental health, and mental health screening instruments.


The attribute rating test for sensory analysis.
This new sensory method evaluates the intensity of an odorant by using a modified "paired-comparison" format. It can be applied to any situation for which the odor-causing agent is known and a purified standard is available. The attribute rating test requires that a standard solution of the odorant be prepared at a concentration that is both readily recognized by most individuals and likely to evoke customer complaints. Within an hour individuals can be trained to compare the odor type and intensity of the standard to water samples, then rate the odor in the sample as "not detected," "less than the standard," "equal to standard," or "greater than the standard." The attribute rating test was successfully used by a full-scale water treatment facility to monitor the intensity of geosmin in the source water and to determine the effectiveness of powdered activated carbon for controlling its earthy-odor in treated water.