The evolution of YidC/Oxa/Alb3 family in the three domains of life: a phylogenomic analysis.
YidC/Oxa/Alb3 family includes a group of conserved translocases that are essential for protein insertion into inner membranes of bacteria and mitochondria, and thylakoid membranes of chloroplasts. Because mitochondria and chloroplasts are of bacterial origin, Oxa and Alb3, like many other mitochondrial/chloroplastic proteins, are hypothetically derived from the pre-existing protein (YidC) of bacterial endosymbionts. Here, we test this hypothesis and investigate the evolutionary history of the whole YidC/Oxa/Alb3 family in the three domains of life. Our comprehensive analyses of the phylogenetic distribution and phylogeny of the YidC/Oxa/Alb3 family lead to the following findings: 1) In archaea, YidC homologs are only sporadically distributed in Euryarchaeota; 2) Most bacteria contain only one YidC gene copy; some species in a few taxa (Bacillus, Lactobacillales, Actinobacteria and Clostridia) have two gene copies; 3) Eukaryotic Oxa and Alb3 have two separate prokaryotic origins, but they might not arise directly from the YidC of proteobacteria and cyanobacteria through the endosymbiosis origins of mitochondrium and chloroplast, respectively; 4) An ancient duplication occurred on both Oxa and Alb3 immediately after their origins, and thus most eukaryotes generally bear two Oxa and two Alb3. However, secondary loss, duplication or acquisition of new domain also occurred on the two genes in some lineages, especially in protists, resulting in a rich diversity or adaptive differentiation of the two translocases in these lineages. YidC is distributed in bacteria and some Euryarchaeota. Although mitochondrial Oxa and chloroplastic Alb3 are derived from the prokaryotic YidC, their origin might be not related to the endosymbiosis events of the two organelles. In some eukaryotic lineages, especially in protists, Oxa and Alb3 have diverse evolutionary histories. Finally, a model for the evolutionary history of the entire YidC/Oxa/Alb3 family in the three domains of life is proposed.


Catalytic activity and isoform-specific inhibition of rat cytochrome p450 4F enzymes.
Arachidonic acid is omega-hydroxylated to 20-hydroxyeicosatetraenoic acid (20-HETE), which has effects on vasoactivity and renal tubular transport and has been implicated in the regulation of blood pressure. Cytochrome p450 (p450) 4A isoforms are generally considered the major arachidonic acid omega-hydroxylases; however, little is known about the role of rat CYP4F isoforms in 20-HETE formation. The rat CYP4F isoforms, CYP4F1, CYP4F4, CYP4F5, and CYP4F6, were heterologously expressed in Escherichia coli, and their substrate specificity in fatty acid metabolism was characterized. Substrate-binding assays indicated that leukotriene B(4) (LTB(4)) and arachidonic acid bound CYP4F1 and CYP4F4 in a type-I manner with a K(s) of 25 to 59 microM, and lauric acid bound CYP4F4 poorly. Reconstituted CYP4F1 and CYP4F4 catalyzed the omega-hydroxylation of LTB(4) with a K(m) of 24 and 31 microM, respectively, and CYP4F5 had minor activity in LTB(4) metabolism. Importantly, CYP4F1 and CYP4F4 catalyzed the omega-hydroxylation of arachidonic acid with an apparent k(cat) of 9 and 11 min(-1), respectively. Lauric acid was a poor substrate for all of the CYP4F isoforms, and CYP4F6 had no detectable fatty acid omega-hydroxylase activity. The p450 omega-hydroxylase inhibitors 17-octadecynoic acid, 10-undecynyl sulfate, and N-methylsulfonyl-12,12-dibromododec-11-enamide showed isoform-specific inhibition of CYP4F1- and CYP4F4-catalyzed omega-hydroxylation of arachidonic acid and potency differences between the CYP4A and CYP4F isoforms. These data support a significant role for CYP4F1 and CYP4F4 in the formation of 20-HETE and identify p450 inhibitors that can be used to understand the relative contribution of the CYP4A and CYP4F isoforms to renal 20-HETE formation.


Electron-microscopic mapping of AT-rich regions and of E. coli RNA polymerase-binding sites on the circular kinetoplast DNA of Trypanosoma cruzi.
Partial alkaline denaturation of the circular kinetoplast DNA (kDNA) of Trypanosoma cruzi has shown the existence of 4 small, well-defined AT-rich regions with an average size of about 200 base pairs. They are almost equally distributed, separated by approximately 90 degrees on the circular molecule. All minicircles, whether free or linked in networks, have the same denaturation pattern and, therefore, seem to contain the same information. The long linear molecules present in low amounts in the kDNA samples do not show the same denaturation pattern. Partial denaturation of molecules in larger associations indicates that the circular units may be linked to each other by one strand only. kDNA can be transcribed in vitro by the RNA polymerase of E. coli. RNA polymerase-kDNA complexes have been studied in the electron microscope. By spreading the DNA-protein complexes by adhesion to positively charged carbon films and dark-field observation, it was possible to show the existence of 4 specific binding sites of the E. coli RNA polymerase on the kDNA circles. Comparing the position of the polymerase-binding sites and the AT-rich melted zones, it is suggested that a correlation exists between the two. As had been shown in earlier work, the replication of circular kDNA can be blocked by treating the trypanosomes with the trypanocidal drug Berenil. The comparison of the relative position of the Berenil-blocked replication forks with the position of the 4 denaturation loops shows that the DNA replication is stopped at these AT-rich regions. Since there is evidence that Berenil binds preferentially to AT-rich DNA and seems to be involved in inhibition of DNA replication, the following hypothetical model can be proposed. The replication of the circular kDNA molecules is discontinuous and involves the synthesis of RNA primers; when Berenil is bound to the AT-rich regions, synthesis of new RNA primers is inhibited and replication is blocked at these points, leading to the accumulation of replicating intermediates with defined branch lengths.


Systematic Cys mutagenesis of FlgI, the flagellar P-ring component of Escherichia coli.
The bacterial flagellar motor is embedded in the cytoplasmic membrane, and penetrates the peptidoglycan layer and the outer membrane. A ring structure of the basal body called the P ring, which is located in the peptidoglycan layer, is thought to be required for smooth rotation and to function as a bushing. In this work, we characterized 32 cysteine-substituted Escherichia coli P-ring protein FlgI variants which were designed to substitute every 10th residue in the 346 aa mature form of FlgI. Immunoblot analysis against FlgI protein revealed that the cellular amounts of five FlgI variants were significantly decreased. Swarm assays showed that almost all of the variants had nearly wild-type function, but five variants significantly reduced the motility of the cells, and one of them in particular, FlgI G21C, completely disrupted FlgI function. The five residues that impaired motility of the cells were localized in the N terminus of FlgI. To demonstrate which residue(s) of FlgI is exposed to solvent on the surface of the protein, we examined cysteine modification by using the thiol-specific reagent methoxypolyethylene glycol 5000 maleimide, and classified the FlgI Cys variants into three groups: well-, moderately and less-labelled. Interestingly, the well- and moderately labelled residues of FlgI never overlapped with the residues known to be important for protein amount or motility. From these results and multiple alignments of amino acid sequences of various FlgI proteins, the highly conserved region in the N terminus, residues 1-120, of FlgI is speculated to play important roles in the stabilization of FlgI structure and the formation of the P ring by interacting with FlgI molecules and/or other flagellar components.


Disarming Burkholderia pseudomallei: structural and functional characterization of a disulfide oxidoreductase (DsbA) required for virulence in vivo.
The intracellular pathogen Burkholderia pseudomallei causes the disease melioidosis, a major source of morbidity and mortality in southeast Asia and northern Australia. The need to develop novel antimicrobials is compounded by the absence of a licensed vaccine and the bacterium's resistance to multiple antibiotics. In a number of clinically relevant Gram-negative pathogens, DsbA is the primary disulfide oxidoreductase responsible for catalyzing the formation of disulfide bonds in secreted and membrane-associated proteins. In this study, a putative B. pseudomallei dsbA gene was evaluated functionally and structurally and its contribution to infection assessed. Biochemical studies confirmed the dsbA gene encodes a protein disulfide oxidoreductase. A dsbA deletion strain of B. pseudomallei was attenuated in both macrophages and a BALB/c mouse model of infection and displayed pleiotropic phenotypes that included defects in both secretion and motility. The 1.9 Å resolution crystal structure of BpsDsbA revealed differences from the classic member of this family Escherichia coli DsbA, in particular within the region surrounding the active site disulfide where EcDsbA engages with its partner protein E. coli DsbB, indicating that the interaction of BpsDsbA with its proposed partner BpsDsbB may be distinct from that of EcDsbA-EcDsbB. This study has characterized BpsDsbA biochemically and structurally and determined that it is required for virulence of B. pseudomallei. These data establish a critical role for BpsDsbA in B. pseudomallei infection, which in combination with our structural characterization of BpsDsbA will facilitate the future development of rationally designed inhibitors against this drug-resistant organism.


The Relationship between Infectious Diseases and Housing Maintenance in Indigenous Australian Households.
This research aimed to identify systemic housing-level contributions to infectious disease transmission for Indigenous Australians, in response to the Government program to 'close the gap' of health and other inequalities. A narrative literature review was performed in accordance to PRISMA guidelines. The findings revealed a lack of housing maintenance was associated with gastrointestinal infections, and skin-related diseases were associated with crowding. Diarrhoea was associated with the state of food preparation and storage areas, and viral conditions such as influenza were associated with crowding. Gastrointestinal, skin, ear, eye, and respiratory illnesses are related in various ways to health hardware functionality, removal and treatment of sewage, crowding, presence of pests and vermin, and the growth of mould and mildew. The research concluded that infectious disease transmission can be reduced by improving housing conditions, including adequate and timely housing repair and maintenance, and the enabling environment to perform healthy behaviours.


Girls start life on an uneven playing field: Evidence from lowland rural Nepal.
Evolutionary research on the sex ratio at birth (SRB) has focused on explaining variability within and between populations, and whether parental fitness is maximized by producing daughters or sons. We tested predictors of SRB in a low-income setting, to understand whether girls differ from boys in their likelihood of being born into families with the capacity to invest in them, which has implications for their future health and fitness. We used data from a cluster randomized control trial from lowland rural Nepal (16 115 mother-child dyads). We applied principal component analysis to extract two composite indices reflecting maternal socio-economic and reproductive (parity, age) capital. We fitted mixed-effects logistic regression models to estimate odds ratios of having a girl in association with these individual factors and indices. The SRB was 112. Compared to the global reference SRB (105), there were seven missing girls per 100 boys. Uneducated, early-marrying, poorer and shorter mothers were more likely to give birth to girls. Analysing composite maternal indices, lower socio-economic and reproductive capital were independently associated with a greater likelihood of having a girl. In this population, girls start life facing composite disadvantages, being more likely than boys to be born to mothers with lower socio-economic status and reproductive capital. Both physiological and behavioural mechanisms may contribute to these epidemiological associations. Differential early exposure by sex to maternal factors may underpin intergenerational cycles of gender inequality, mediated by developmental trajectory, education and socio-economic status.


Interim Report of a Prospective Trial on the Clinical Efficiency of a New Full-thickness Endoscopic Plication Device for Patients With GERD: Impact of Changed Suture Material.
The aim of the present trial is to investigate the clinical efficiency of the GERDx device for patients with gastroesophageal reflux disease (GERD). Prospective study evaluating Gastrointestinal Quality of Life Index, symptoms scores, as well as esophageal manometry and impedance-pH-monitoring data at baseline and 3 months after endoscopic full-thickness plication with the GERDx device. In total, 28 patients underwent the procedure so far. Mean Gastrointestinal Quality of Life Index scores, DeMeester scores, and general and reflux-specific scores improved (P<0.01). Three of 6 patients, who were treated with converted sutures, experienced postoperative complications. Endoscopic plication using the GERDx device may be effective in improving quality of life and GERD symptoms. Suture length between pledgets and suture material may have an impact on procedure outcomes.


New approaches in the management of multiple sclerosis.
Multiple sclerosis (MS) is a central nervous system chronic inflammatory disease that is characterized by an extensive and complex immune response. Scientific advances have occurred in immunology, pathophysiology, and diagnostic and clinical assessment tools, and recent discovery of unique therapeutic targets has spurred numerous Phase II and Phase III clinical trials. Reductions in MS relapse rates and improvements in T₂ or gadolinium-enhancing lesion burdens have been reported from Phase III trials that include fingolimod, alemtuzumab, cladribine, and rituximab. Promising Phase II trial data exist for teriflunomide, daclizumab, laquinimod, and fumarate. The optimism created by these favorable findings must be tempered with evaluation of the adverse effect profile produced by these new agents. Given the discovery of progressive multifocal leukoencephalopathy with the use of natalizumab, ongoing vigilance for rare and life-threatening reactions due to new agents should be paramount. Patients with MS often experience difficulty with ambulation, spasticity, and cognition. Recent clinical trial data from two Phase III dalfampridine-SR trials indicate certain patients receive benefits in ambulation. This article provides an overview of data from clinical trials of newer agents of potential benefit in MS.


Self-trapping of polychromatic light in nonlinear periodic photonic structures.
We study dynamical reshaping of polychromatic beams due to collective nonlinear self-action of multiple-frequency components in periodic and semi-infinite photonic lattices and predict the formation of polychromatic gap solitons and polychromatic surface solitons due to light localization in spectral gaps. We show that the self-trapping efficiency and structure of emerging polychromatic solitons depend on the input spectrum due to the lattice-enhanced dispersion, including the effect of crossover from localization to diffraction in media with defocusing nonlinearity.


Amyloid-related Imaging Abnormalities in Alzheimer Disease Treated with Anti-Amyloid-β Therapy.
Alzheimer disease (AD) is the most common form of dementia worldwide. Treatment of AD has mainly been focused on symptomatic treatment until recently with the advent and approval of monoclonal antibody (MAB) immunotherapy. U.S. Food and Drug Administration-approved drugs such as aducanumab, as well as upcoming newer-generation drugs, have provided an exciting new therapy focused on reducing the amyloid plaque burden in AD. Although this new frontier has shown benefits for patients, it is not without complications, which are mainly neurologic. Increased use of MABs led to the discovery of amyloid-related imaging abnormalities (ARIA). ARIA has been further classified into two categories, ARIA-E and ARIA-H, representing edema and/or effusion and hemorrhage, respectively. ARIA is thought to be caused by increased vascular permeability following an inflammatory response, leading to the extravasation of blood products and proteinaceous fluid. Patients with ARIA may present with headaches, but they are usually asymptomatic and ARIA is only diagnosable at MRI; it is essential for the radiologist to recognize and monitor ARIA. Increased incidence and investigation into this concern have led to the creation of grading scales and monitoring guidelines to diagnose and guide treatment using MABs. Cerebral amyloid angiopathy has an identical pathogenesis to that of ARIA and is its closest differential diagnosis, with imaging findings being the same for both entities and only a history of MAB administration allowing differentiation. The authors discuss the use of MABs for treating AD, expand on ARIA and its consequences, and describe how to identify and grade ARIA to guide treatment properly. ©RSNA, 2023 Quiz questions for this article are available through the Online Learning Center See the invited commentary by Yu in this issue.


[Sucrose utilization determinants of transposon Tn2555].
The Sac- -derivatives of the plasmid pBRS5.2 containing the sucrose utilization transposon Tn2555 were obtained by using the insertional mutagenesis. The sac mutations were mapped within the transposons DNA fragment of about 3kb. The existence of two linkage groups of the sac mutations was shown by complementation analysis. Among the gene products of Tn2555 the polypeptides of about 58, 36, 27, 14 and 12 kd were found. The data on relation of the sac genes of Tn2555 and the ones of the known plasmid pUR400 of Salmonella typhimurium are discussed.


Fibre-Reinforced Polymer Reinforced Concrete Members under Elevated Temperatures: A Review on Structural Performance.
Several experimental and numerical studies have been conducted to address the structural performance of FRP-reinforced/strengthened concrete structures under and after exposure to elevated temperatures. The present paper reviews over 100 research studies focused on the structural responses of different FRP-reinforced/strengthened concrete structures after exposure to elevated temperatures, ranging from ambient temperatures to flame. Different structural systems were considered, including FRP laminate bonded to concrete, FRP-reinforced concrete, FRP-wrapped concrete, and concrete-filled FRP tubes. According to the reported data, it is generally accepted that, in the case of insignificant resin in the post curing process, as the temperature increases, the ultimate strength, bond strength, and structure stiffness reduce, especially when the glass transition temperature Tg of the resin is approached and exceeded. However, in the case of post curing, resin appears to preserve its mechanical properties at high temperatures, which results in the appropriate structural performance of FRP-reinforced/strengthened members at high temperatures that are below the resin decomposition temperature Td. Given the research gaps, recommendations for future studies have been presented. The discussions, findings, and comparisons presented in this review paper will help designers and researchers to better understand the performance of concrete structures that are reinforced/strengthened with FRPs under elevated temperatures and consider appropriate approaches when designing such structures.


Interstitial High-Dose-Rate Gynecologic Brachytherapy: Clinical Workflow Experience From Three Academic Institutions.
An interstitial brachytherapy approach for gynecologic cancers is typically considered for patients with lesions exceeding 5 mm within tissue or that are not easily accessible for intracavitary applications. Recommendations for treating gynecologic malignancies with this approach are available through the American Brachytherapy Society, but vary based on available resources, staffing, and logistics. The intent of this manuscript is to share the collective experience of 3 academic centers that routinely perform interstitial gynecologic brachytherapy. Discussion points include indications for interstitial implants, procedural preparations, applicator selection, anesthetic options, imaging, treatment planning objectives, clinical workflows, timelines, safety, and potential challenges. Interstitial brachytherapy is a complex, high-skill procedure requiring routine practice to optimize patient safety and treatment efficacy. Clinics planning to implement this approach into their brachytherapy practice may benefit from considering the discussion points shared in this manuscript.


Prostate surgeries: average charges throughout the United States, 1997.
During 1997 hospital claims for 2,149 radical (open) prostatectomies among selected group policy insureds averaged $16,990 and for 2,192 transurethral resections of the prostate (TURPs), $6,620. There was marked geographic variation in both the distribution and charges for these two procedures. The Pacific region reported the highest average total charges for both procedures, driven by the high charges in California. The California average total charges were 39 percent above the norm for an open procedure and 54 percent above for a TURP. Illinois and Florida each reported open prostatectomy charges over $20,000, 20 and 18 percent, respectively, above the norm. For a TURP, however, Minnesota and Arizona reported the second and third highest charges, each over $8,000, and around 30 percent above the average. Washington state reported the lowest average charge for an open procedure ($12,020, 29 percent below the norm and about half that in California); Pennsylvania had the lowest TURP total charge ($3,860, 42 percent below the norm and 62 percent lower than the California charge). Among study states, charges 15 percent or more below average for an open procedure were also recorded in Pennsylvania, Indiana, Maryland and North Carolina. Similarly low charges for a TURP were recorded in Ohio. Michigan and North Carolina (each more than 15 percent below the norm). The hospital charges comprised 63 percent of the total for an open procedure and 68 percent for a TURP. The California average was the highest for both surgeries. 58 and 69 percent, respectively, above the norm and 129 percent above the low charge in Maryland for an open procedure and 193 percent above the low charge in Pennsylvania for a TURP. Physicians' charges averaged $6,370 for an open prostatectomy and $2,130 for a TURP. For both surgeries, these charges were the highest in New Jersey and New York (between 20 and 30 percent above the norm) and the lowest in Pennsylvania where they averaged 28 percent lower than the norm for an open procedure and 40 percent lower for a TURP. The length of stay averaged 3.75 days for a radical prostatectomy and 2.80 days for a TURP. These days ranged from 4.20 days in New Jersey to 3.11 days in Minnesota for an open procedure and 3.51 days in New Jersey to 2.14 days in Minnesota for a TURP.


Evidence to suggest that expression of MITF induces melanocyte differentiation and haploinsufficiency of MITF causes Waardenburg syndrome type 2A.
MITF (microphthalmia-associated transcription factor) encodes a transcription factor with a basic-helix-loop-helix-leucine zipper (bHLH-Zip) motif. Ectopic expression of MITF is found to convert NIH/3T3 fibroblasts into cells with characteristics of melanocytes. MITF transfectants formed foci, which superficially resembled those induced by oncogenes, but did not exhibit malignant phenotypes. Instead, they contained dendritic cells that express melanogenic marker proteins such as tyrosinase and tyrosinase-related protein 1. Such properties were not observed in cells transfected with the closely related gene, TFE3. These findings indicated that MITF is involved in melanocyte differentiation. Two mutations (C760-->T and C895-->T) in MITF are found to be associated with individuals with Waardenburg syndrome type 2 (WS2). These mutations create stop codons in exon 7 and 8, respectively, and probably result in truncated proteins lacking HLH-Zip or Zip structure. To understand how these MITF mutations cause WS2 in heterozygotes, mutant MITF proteins were generated and used for DNA-binding and luciferase reporter assays. The mutated MITF proteins lose their DNA-binding activity and fail to transactivate the promoter of the tyrosinase gene. However, these mutated proteins do not appear to interfere with the activity of wild-type MITF protein in these assays, indicating that they do not show a dominant-negative effect. These findings suggest that the phenotypes of the two WS2 families are caused by loss-of-function mutations in one of the two MITF alleles, resulting in haploinsufficiency of the MITF protein, the transcription factor necessary for normal melanocyte differentiation.


Leukocyte survival in cerebrospinal fluid.
Delays in the laboratory examination of cerebrospinal fluid are commonly encountered in clinical medicine. The present studies were designed to evaluate changes in cerebrospinal fluid leukocyte counts relative to time elapsed before analysis. Neutrophil counts decreased most rapidly, being 68 +/- 10% (standard error of the mean) and 50 +/- 12% of initial values at 1 and 2 h, respectively. Lymphocyte and monocyte numbers were not significantly altered until 3 h.


Negative-stain electron microscopy of inside-out FtsZ rings reconstituted on artificial membrane tubules show ribbons of protofilaments.
FtsZ, the primary cytoskeletal element of the Z ring, which constricts to divide bacteria, assembles into short, one-stranded filaments in vitro. These must be further assembled to make the Z ring in bacteria. Conventional electron microscopy (EM) has failed to image the Z ring or resolve its substructure. Here we describe a procedure that enabled us to image reconstructed, inside-out FtsZ rings by negative-stain EM, revealing the arrangement of filaments. We took advantage of a unique lipid that spontaneously forms 500 nm diameter tubules in solution. We optimized conditions for Z-ring assembly with fluorescence light microscopy and then prepared specimens for negative-stain EM. Reconstituted FtsZ rings, encircling the tubules, were clearly resolved. The rings appeared as ribbons of filaments packed side by side with virtually no space between neighboring filaments. The rings were separated by variable expanses of empty tubule as seen by light microscopy or EM. The width varied considerably from one ring to another, but each ring maintained a constant width around its circumference. The inside-out FtsZ rings moved back and forth along the tubules and exchanged subunits with solution, similarly to Z rings reconstituted outside or inside tubular liposomes. FtsZ from Escherichia coli and Mycobacterium tuberculosis assembled rings of similar structure, suggesting a universal structure across bacterial species. Previous models for the Z ring in bacteria have favored a structure of widely scattered filaments that are not in contact. The ribbon structure that we discovered here for reconstituted inside-out FtsZ rings provides what to our knowledge is new evidence that the Z ring in bacteria may involve lateral association of protofilaments.


Interphase Regulation by Multifunctional Additive Empowering High Energy Lithium-Ion Batteries with Enhanced Cycle Life and Thermal Safety.
High energy density lithium-ion batteries (LIBs) adopting high-nickel layered oxide cathodes and silicon-based composite anodes always suffer from unsatisfied cycle life and poor safety performance, especially at elevated temperatures. Electrode/electrolyte interphase regulation by functional additives is one of the most economic and efficacious strategies to overcome this shortcoming. Herein, cyano-groups (-CN) are introduced into lithium fluorinated phosphate to synthesize a novel multifunctional additive of lithium tetrafluoro (1,2-dihydroxyethane-1,1,2,2-tetracarbonitrile) phosphate (LiTFTCP), which endows high nickel LiNi0.8Co0.1Mn0.1O2 /SiOx-graphite composite full cell with an ultrahigh cycle life and superior safety characteristics, by adding only 0.5 wt.% LiTFTCP into a LiPF6-carbonate baseline electrolyte. It is revealed that LiTFTCP additive effectively suppresses the HF generation and facilitates the formation of a robust and heat-resistant cyano-enriched CEI layer as well as a stable LiF-enriched SEI layer. The favorable SEI/CEI layers greatly lessen the electrode degradation, electrolyte consumption, thermal-induced gassing and total heat-releasing. This work illuminates the importance of additive molecular engineering and interphase regulation in simultaneously promoting the cycling and thermal safety of LIBs with high-nickel NCMxyz cathode and silicon-based composite anode.


Proteomic and phosphoproteomic analysis of responses to enterovirus A71 infection reveals novel targets for antiviral and viral replication.
Hand, foot, and mouth disease (HFMD) is a common infectious disease in infants and children, especially those under five years of age. EV-A71 is a common pathogen that causes HFMD and the primary pathogen leading to severe or fatal HFMD, which is characterized by neurological complications. However, the underlying mechanisms of EV-A71 pathogenesis remain largely unknown. In this report, we used proteomic and phosphorylated proteomic methods to characterize the proteome and phosphoproteome profiles of EV-A71-infected human neuroblastoma SK-N-SH cells. More than 7744 host proteins and 10069 phosphorylation modification sites were successfully quantified. Among them, 974 proteins and 3648 phosphorylation modification sites were regulated significantly during EV-A71 infection. KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis revealed that EV-A71 altered cell biological processes, including protein synthesis, RNA splicing and metabolism in SK-N-SH cells. Notably, based on the prediction of upregulated kinases during EV-A71 infection, we identified specific kinase inhibitors approved by the FDA, with ceralasertib, bosutinib, flavin mononucleotide, minocycline, pimasertib and acetylcysteine inhibiting EV-A71 infection. Finally, EV-A71 proteins were found to be phosphorylated during infection, with one site (S184 on 3D polymerase) observed to be crucial for viral replication because a S184A mutation knocked out viral replication. The results improve our understanding of the host response to EV-A71 infection of neuroblastoma cells and provide potential targets for developing anti-EV-A71 strategies.


Spared nerve injury decreases motivation in long-access homecage-based operant tasks in mice.
Neuropathic pain causes both sensory and emotional maladaptation. Preclinical animal studies of neuropathic pain-induced negative affect could result in novel insights into the mechanisms of chronic pain. Modeling pain-induced negative affect, however, is variable across research groups and conditions. The same injury may or may not produce robust negative affective behavioral responses across different species, strains, and laboratories. Here, we sought to identify negative affective consequences of the spared nerve injury model on C57BL/6J male and female mice. We found no significant effect of spared nerve injury across a variety of approach-avoidance conflict, hedonic choice, and coping strategy assays. We hypothesized these inconsistencies may stem in part from the short test duration of these assays. To test this hypothesis, we used the homecage-based Feeding Experimentation Device version 3 to conduct 12-hour, overnight progressive ratio testing to determine whether mice with chronic spared nerve injury had decreased motivation to earn palatable food rewards. Our data demonstrate that despite equivalent task learning, spared nerve injury mice are less motivated to work for a sugar pellet than sham controls. Furthermore, when we normalized behavioral responses across all the behavioral assays we tested, we found that a combined normalized behavioral score is predictive of injury state and significantly correlates with mechanical thresholds. Together, these results suggest that homecage-based operant behaviors provide a useful platform for modeling nerve injury-induced negative affect and that valuable pain-related information can arise from agglomerative data analyses across behavioral assays-even when individual inferential statistics do not demonstrate significant mean differences.


Isolated IgG4-related gastric disease presenting as diffuse gastric wall thickening with ulcer.
An 81-year-old male presented with loss of appetite, early satiety and iron deficiency anaemia. A computed tomography (CT) scan of the abdomen and pelvis during initial work-up revealed diffuse gastric mural thickening associated with a large ulcer and adjacent gastro-hepatic lymphadenopathy. The CT appearances, together with the clinical features, were highly suspicious for an infiltrative type of gastric malignancy. Endoscopic biopsy however showed erosive inflammation, IgG4 plasmacytosis and fibrosis, raising the possibility of IgG4-related disease. A serologic assay for IgG showed normal IgG4 and elevated IgG2 serum levels. After appropriate steroid treatment, endoscopy and CT scan showed resolution of the ulcer and gastric wall thickening. This case shows yet another possible appearance of gastric involvement in IgG4-related disease on the current evolving spectrum of this disease presentation. Greater awareness and education of this disease would help in patient care, ensuring earlier diagnosis, prevention of severe organ damage and morbidity, as well as unnecessary surgery.


Clinical deficits correlate with regional cerebral atrophy in progressive supranuclear palsy.
Most cerebral imaging studies of patients with progressive supranuclear palsy (PSP) have noted subtle atrophy, although the full extent of atrophy and any correlates to clinical features have not been determined. We used voxel-based morphometry analysis of grey matter, white matter and CSF on MRI brain scans to map the statistical probability of regional tissue atrophy in 21 patients with PSP, 17 patients with Parkinson's disease and 23 controls. PSP and Parkinson's disease cohorts were selected to approximate the mid-stages of their respective disease courses. Where regions of significant tissue atrophy were identified in a disease group relative to controls, the probability of tissue loss within those regions was correlated with global indices of motor disability, and behavioural and cognitive disturbance for that disease group. Minimal regional atrophy was observed in Parkinson's disease. PSP could be distinguished from both controls and Parkinson's disease by symmetrical tissue loss in the frontal cortex (maximal in the orbitofrontal and medial frontal cortices), subcortical nuclei (midbrain, caudate and thalamic) as well as periventricular white matter. For PSP, motor deficits correlated with atrophy of the caudate and motor cingulate, while behavioural changes related to atrophy in the orbitofrontal cortex and midbrain. These data suggest that intrinsic neurodegeneration of specific subcortical nuclei and frontal cortical subregions together contribute to motor and behavioural disturbances in PSP and differentiate this disorder from Parkinson's disease within 2-4 years of symptom onset.


Tolerance of staphylococcal thermonuclease to stress.
Remarkable tolerance to prolonged heating, prolonged storage, and bacterial proliferation was exhibited by staphylococcal thermonuclease in foods and broth. A purified enzyme preparation added to Brain Heart Infusion broth was unaffected by the growth of five bacterial species. Minimal inactivation was effected by Bacillus subtilis. Optimal growth of Streptococcus faecalis var. liquifaciens caused extensive inactivation of thermonuclease. However, storage at room temperature or the addition of 5% NaCl caused only minimal inactivation.


Tricarboxylic acid cycle-dependent regulation of Staphylococcus epidermidis polysaccharide intercellular adhesin synthesis.
Staphylococcus epidermidis is a major nosocomial pathogen primarily infecting immunocompromised individuals or those with implanted biomaterials (e.g., catheters). Biomaterial-associated infections often involve the formation of a biofilm on the surface of the medical device. In S. epidermidis, polysaccharide intercellular adhesin (PIA) is an important mediator of biofilm formation and pathogenesis. Synthesis of PIA is regulated by at least three DNA binding proteins (IcaR, SarA, and sigma(B)) and several environmental and nutritional conditions. Previously, we observed the environmental conditions that increased PIA synthesis decreased tricarboxylic acid (TCA) cycle activity. In this study, S. epidermidis TCA cycle mutants were constructed, and the function of central metabolism in PIA biosynthesis was examined. TCA cycle inactivation altered the metabolic status of S. epidermidis, resulting in a massive derepression of PIA biosynthetic genes and a redirection of carbon from growth into PIA biosynthesis. These data demonstrate that the bacterial metabolic status is a critical regulatory determinant of PIA synthesis. In addition, these data lead us to propose that the TCA cycle acts as a signal transduction pathway to translate external environmental cues into intracellular metabolic signals that modulate the activity of transcriptional regulators.


Tumor necrosis factor-alpha stimulates superoxide anion generation by perfused rat liver and Kupffer cells.
Tumor necrosis factor (TNF) has been implicated as one of the mediators of the immunologic and metabolic changes in endotoxemia. Under adverse conditions, TNF can also be cytotoxic, and its effects can ultimately contribute to organ failure. This study shows that a 30-min infusion of a nonlethal dose of TNF induced the release of superoxide anion (0.9 nmol.min-1.g-1) by the in situ perfused rat liver. TNF also primed the liver to generate more superoxide anion (2.0 nmol.min-1.g-1) in response to an in vitro challenge with phorbol 12-myristate 13-acetate (PMA). Kupffer cells are most likely responsible for the superoxide anion production under these conditions, because the isolated Kupffer cells from TNF-infused rats produced increased quantities of superoxide anion (4-8 nmol/10(6) cells) when subsequently treated in vitro with either PMA or opsonized zymosan (control less than 1 nmol/10(6) cells). Thus, under these experimental conditions, TNF in vivo primed the Kupffer cells, but not the hepatocytes, endothelial cells, and the blood or hepatic neutrophils, to release more superoxide anion. These studies indicate that during a short-term nonlethal TNF infusion, Kupffer cells are a major target of TNF action, leading to the release of toxic-oxygen metabolites that may contribute to organ failure.


Achieving arsenic concentrations of <1 μg/L by Fe(0) electrolysis: The exceptional performance of magnetite.
Consumption of drinking water containing arsenic at concentrations even below the World Health Organization provisional limit of 10 μg/L can still lead to unacceptable health risks. Consequently, the drinking water sector in the Netherlands has recently agreed to target 1 μg/L of arsenic in treated water. Unfortunately, in many poor, arsenic-affected countries, the costs and complexity of current methods that can achieve <1 μg/L are prohibitive, which highlights the need for innovative methods that can remove arsenic to <1 μg/L without costly support infrastructure and complicated supply chains. In this work, we used Fe(0) electrolysis, a low cost and scalable technology that is also known as Fe(0) electrocoagulation (EC), to achieve <1 μg/L residual dissolved arsenic. We compared the arsenic removal performance of green rust (GR), ferric (oxyhydr)oxides (Fe(III) oxides) and magnetite (Mag) generated by EC at different pH (7.5 and 9) in the presence of As(III) or As(V) (initial concentrations of 200-11,000 μg/L). Although GR and Fe(III) oxides removed up to 99% of initial arsenic, neither Fe phase could reliably meet the 1 μg/L target at both pH values. In contrast, EC-generated Mag consistently achieved <1 μg/L, regardless of the initial As(V) concentration and pH. Only solutions with initial As(III) concentrations ≥2200 μg/L resulted in residual arsenic >1 μg/L. As K-edge X-ray absorption spectroscopy showed that Mag also sorbed arsenic in a unique mode, consistent with partial arsenic incorporation near the particle surface. This sorption mode contrasts with the binuclear, corner sharing surface complex for GR and Fe(III) oxides, which could explain the difference in arsenic removal efficiency among the three Fe phases. Our results suggest that EC-generated Mag is an attractive method for achieving <1 μg/L particularly in decentralized water treatment.


The keratitis, ichthyosis, and deafness (KID) syndrome.
We describe here a patient with ichthyosis, keratitis, deafness, and recalcitrant cutaneous bacterial and fungal infections, who was previously described as having hereditary hypohidrotic ectodermal dysplasia. Similarly affected patients described in the literature have the following features in common: (1) a distinctive ichthyosis characterized by a fine dry scale, follicular hyperkeratotic spines, and a reticulated pattern of hyperkeratosis on the palms and soles; (2) a vascularizing keratitis that results in notable visual impairment; and (3) neurosensory deafness. One half of the affected patients also display frequent, severe cutaneous infections. We suggest that the name "the KID syndrome" to emphasize the characteristic features of the syndrome-keratitis, ichthyosis, and deafness.


Trauma-hemorrhagic shock mesenteric lymph induces endothelial apoptosis that involves both caspase-dependent and caspase-independent mechanisms.
To determine the mechanism by which gut-derived factors present in mesenteric lymph from rats subjected to trauma-hemorrhagic shock (T/HS) induce endothelial cell death. Intestinal ischemia after hemorrhagic shock results in gut barrier dysfunction and the subsequent production of biologically active and tissue injurious factors by the ischemic gut. These factors are carried in the mesenteric lymph and reach the systemic circulation via the mesenteric lymph, thereby ultimately resulting in distant organ injury. Although studies have established that trauma-hemorrhagic (T/HS) shock but not trauma-sham-shock (T/SS) mesenteric lymph is cytotoxic to endothelial cells, whether T/HS lymph-induced endothelial cell death occurs via an apoptotic or a necrotic pathway is unknown. The mechanisms underlying T/HS lymph-induced cytotoxicity are likewise unknown. Human umbilical vein endothelial cell (HUVEC) monolayers were incubated with medium, sham-shock, or post shock mesenteric lymph (5%) for 4 hours, after which the mode of cell death (ie, apoptosis versus necrosis) was determined using morphologic (confocal microscopy), biochemical (nucleosomal release), and DNA-based (gel electrophoresis) assays. To clarify the cellular pathways involved in T/HS lymph-induced HUVEC cell death, caspase-3, caspase-9, caspase-8, and BID activity was measured as was the ability of the pan-caspase inhibitor z-VAD-fmk to prevent T/HS lymph-induced cell death. T/HS, but not T/SS, mesenteric lymph or medium was cytotoxic and caused the appearance of the classic morphologic signs of apoptosis, including membrane blebbing, cell shrinkage, and apoptotic body formation. Nucleosomal release and a DNA laddering pattern was also observed in the HUVECs incubated with T/HS lymph. These signs of apoptosis were associated with increased caspase activity as reflected in activation of the pro-apoptotic caspases, caspase-8, -9, and -3, as well as the pro-apoptotic bcl-2-related protein BID. However, since the broad-spectrum caspase inhibitor z-VAD-fmk delayed T/HS lymph-induced HUVEC cell death, but did not prevent it fully, it appears that other factors besides caspases are involved in the endothelial cell toxicity of T/HS lymph. Gut-derived factors in T/HS, but not T/SS, mesenteric lymph cause endothelial cell death via an apoptotic mechanism that involves both caspase-dependent and caspase-independent pathways.


[Taurine and the adenosine cyclic monophosphate levels in the heart].
The mechanism of the decrease in the heart of cAMP content under the effect of taurine was studied. It was shown on a preparation of cytoplasmic membranes isolated from the rat heart that under the effect of taurine (10(-6)--10(-3) M) the activity of phosphodiesterase and adenylate cyclase increases to the maximum by 2.4 and 1.4 times, respectively. Taurine weakens the activation of adenylate cyclase by adrenaline. The effect of taurine on adenylate cyclase activity diminishes under the action of phentolamine and obsidan and is practically not manifested in the simultaneous presence of both adrenoblocking agents. Taurine obviously changes the content of cAMP in the heart by altering the activity of adenylate cyclase and phosphodiesterase.


Maintenance of D-alanine ester substitution of lipoteichoic acid by reesterification in Staphylococcus aureus.
Toluene-treated Staphylococcus aureus cells did not synthesize teichoic acid and lipoteichoic acid under the conditions used. The organism displayed, however, a high capacity of incorporating D-[14C]alanine into previously formed polymers. The reaction was dependent on ATP and enhanced by magnesium ions. The incorporation rate into lipoteichoic acid correlated with the rate of loss of alanine ester which occurred through transfer to teichoic acid and base-catalyzed hydrolysis. At pH 6.5 the loss (20% within 4 h) was completely compensated for by reesterification. At pH 7.5 the loss was 60%, but by accelerated incorporation it was reduced to 10%. Incorporation was also enhanced when the original substitution of lipoteichoic acid was lowered by previous growth of S. aureus at high salt concentration. The newly added alanine was randomly distributed along the poly(glycerophosphate) chain. The decreased alanine substitution of lipoteichoic acid after growth at high salt concentration was shown to result from a direct inhibition of alanine incorporation.


Correlation of Retinal Structure and Visual Function Assessments in Mouse Diabetes Models.
Diabetic retinopathy results in vision loss with changes to both retinal blood vessels and neural retina. Recent studies have revealed that animal models of diabetes demonstrate early loss of visual function. We explored the time course of retinal change in three different mouse models of diabetes in a longitudinal study using in vivo measures of retinal structure (optical coherence tomography [OCT]) and visual function (optomotor and pupillary responses). OCT analysis of retinal microstructure, optokinetic response as a measure of visual acuity, and pupillary response to light stimulation were compared among the db/db, Ins2Akita, and streptozotocin (STZ)-induced mouse models of diabetes at 1.5, 3, 6, and 9 months of diabetes. The db/db, Ins2Akita, and STZ-induced models of diabetes all exhibited vision loss and retinal thinning as disease progressed. Both structural changes and functional measures were significantly correlated with the blood glucose levels. Despite this, vision loss and retinal thinning were not consistently correlated, except for the inner retinal layer thickness at 6 months of diabetes. This longitudinal study compiled structural measures and functional outcome data for type 1 and 2 diabetes mouse models commonly used for diabetes studies and demonstrated an overall decline in retinal-related health in conjunction with weight change and blood glucose alterations. The relationship between the structural change and functional outcome could be correlative but is not necessarily causative, as retinal thinning was not sufficient to explain visual acuity decline.


Continuous epidural analgesia for vaginal delivery in Sweden. Report of a nationwide inquiry.
Volumetric pump infusion of bupivacaine into the epidural space for pain relief during labour was introduced in Sweden in 1976. Since then the method has gained widespread use. A questionnaire concerning the use of epidural analgesia (EDA) in 1982 was sent to anaesthetists at all Swedish maternity clinics. During the year, 84 188 vaginal deliveries were registered. EDA for pain relief was performed in 11 324 of them (13.4%). At that time, EDA was available in 56 out of 66 clinics. In almost all clinics (52/56), bupivacaine plain 2.5 mg/ml was used for the blocks. Analgesia was maintained by volumetric pump infusion of bupivacaine in 28 units, by intermittent technique in 25, and both techniques were used in three units. No clinic had abandoned the infusion technique once it had been introduced. Experience from both techniques showed the infusion technique to be more convenient as it produces stable analgesia, is easily conducted and can be supervised by midwives. No complication which could be connected with infusion technique was noted. Maternal hypotension was rare and the instrumental delivery rate was low. The technique proved to be a valuable aid to the organisation of a 24-h service for obstetric analgesia, without requiring any increase in staff.


Heart rate deceleration after exercise predicts patients most likely to respond to cardiac resynchronisation therapy.
This study examines the relationship between heart rate recovery following exercise and subsequent response to cardiac resynchronisation therapy (CRT). Blunted heart rate recovery is an adverse prognostic marker in heart failure and has been shown to correlate with disease severity. 37 patients receiving biventricular pacemakers for conventional indications underwent functional assessments; cardiopulmonary exercise test, 6-min walk test and quality-of-life assessment, together with echo analyses, before and at 3 months following implant. Heart rate deceleration (HRD) gradients were calculated at 30-, 60-, 90- and 120-s intervals following cessation of the baseline exercise test and compared with subsequent markers of response to CRT. Functional response was defined as > or =20% improvement in any two of the three functional assessments, and echo response defined as > or =5% increase in ejection fraction. Functional responders demonstrated steeper HRD gradients than non-responders at 30, 60 and 90 s. Echo responders also demonstrated steeper HRD at 30 and 60 s from the cessation of exercise. Receiver-operating curve analysis demonstrates area under the curve of 0.87 and 0.82, respectively, for HRD30 to predict functional and echo response to CRT. A cut-off value of 3 for HRD30, equating to a 5% reduction in HR between peak exercise and 30 s into recovery, demonstrates the optimal sensitivity/specificity profile to perform this function. HRD following exercise correlates with functional and echocardiographic response to CRT. Application of this parameter in addition to standard criteria may provide valuable supplementary information in the evaluation of prospective CRT candidates.


Dynamics of functional connectivity at high spatial resolution reveal long-range interactions and fine-scale organization.
Dynamic functional connectivity (dFC) derived from resting-state functional magnetic resonance imaging sheds light onto moment-to-moment reconfigurations of large-scale functional brain networks. Due to computational limits, connectivity is typically computed using pre-defined atlases, a non-trivial choice that might influence results. Here, we leverage new computational methods to retrieve dFC at the voxel level in terms of dominant patterns of fluctuations, and demonstrate that this new representation is informative to derive meaningful brain parcellations, capturing both long-range interactions and fine-scale local organization. Specifically, voxelwise dFC dominant patterns were captured through eigenvector centrality followed by clustering across time/subjects to yield most representative dominant patterns (RDPs). Voxel-wise labeling according to positive/negative contributions to RDPs, led to 37 unique labels identifying strikingly symmetric dFC long-range patterns. These included 449 contiguous regions, defining a fine-scale parcellation consistent with known cortical/subcortical subdivisions. Our contribution provides an alternative to obtain a whole-brain parcellation that is for the first time driven by voxel-level dFC and bridges the gap between voxel-based approaches and graph theoretical analysis.


Conjoint Tendon Release in Direct Anterior Total Hip Arthroplasty: No Impact on Patient Outcomes.
The direct anterior (DA) approach for total hip arthroplasty (THA) has become increasingly popular. However, femoral access can be more technically challenging in difficult cases as compared with other approaches. Conjoint tendon release can improve proximal femoral exposure, but its effect on patient function and pain has not been studied. This study evaluated the effect of conjoint tendon release on length of stay (LOS), inpatient pain medication requirements, and functional outcomes of patients undergoing DA THA. The authors retrospectively reviewed charts of all primary DA THAs conducted by a single surgeon between August 2012 and July 2015. Patient demographics, bilateral THA cases, intraoperative conjoint tendon or other soft tissue releases, intraoperative complications, LOS, and inpatient pain medication data were evaluated. One-year functional outcome scores, including the Western Ontario and McMaster Universities Osteoarthritis Index and the Harris Hip Scores, were also reviewed. The authors identified 312 primary DA THAs, with 29 concurrent bilateral THAs. The conjoint tendon was released in 180 cases, whereas a tensor fascia lata (TFL) was released for 29 cases. Mean LOS was 1.3±1.1 days, with patient age (P=.002), bilateral THA (P<.001), TFL release (P=.005), and intraoperative complications (P=.002) predictive of LOS. Mean daily morphine equivalent dose narcotic use was 43.2±48.2 mg, with age being a negative predictor of narcotic use (P=.019). Conjoint tendon release was not predictive of LOS, inpatient pain medication requirements, or outcome scores. Given that conjoint release improves femoral exposure but does not affect LOS or 1-year patient-reported outcomes, intraoperative thresholds for conjoint release should be low. [Orthopedics. 2017; 40(6):e971-e974.].


Lactation and fertility.
In almost all mammals lactation, or more correctly the suckling stimulus, induces a period of infertility designed to provide the optimal birth spacing for survival of the offspring. The duration of lactational infertility depends on the sucking activity of the young with little evidence to support a role for nutritional status. Suckling disrupts the normal pulsatile pattern of hypothalamic gonadotrophin releasing hormone (GnRH)2 secretion resulting in reduced LH secretion from the pituitary. Secretion of FSH returns to its normal cyclic pattern early in lactation and ovarian follicles may develop under its influence. However, until suckling declines, the follicles fail to secrete amounts of estradiol adequate to stimulate an LH surge and ovulation. The suckling stimulus may affect GnRH secretion by affecting prolactin, opiate and dopaminergic tone in the hypothalamus but no unifying mechanism has yet been proposed convincingly. The birth spacing effects of breastfeeding in women have a profound effect on infant well-being, and breastfeeding still prevents more pregnancies than all forms of artificial contraception.


Value of information analysis in environmental health risk management decisions: past, present, and future.
Experts agree that value of information (VOI) analyses provide useful insights in risk management decisions. However, applications in environmental health risk management (EHRM) remain largely demonstrative thus far because of the complexity in modeling and solving VOI problems. Based on this comprehensive review of all VOI applications published in the peer-reviewed literature of such applications, the complexity of solving VOI problems with continuous probability distributions as inputs in models emerges as the main barrier to greater use of VOI although simulation allows analysts to solve more complex and realistic problems. Several analytical challenges that inhibit greater use of VOI techniques include issues related to modeling decisions, valuing outcomes, and characterizing uncertain and variable model inputs appropriately. This comprehensive review of methods for modeling and solving VOI problems for applications related to EHRM provides the first synthesis of important methodological advances in the field. The insights provide risk analysts and decision scientists with some guidance on how to structure and solve VOI problems focused on evaluating opportunities to collect better information to improve EHRM decisions. They further suggest the need for some efforts to standardize approaches and develop some prescriptive guidance for VOI analysts similar to existing guidelines for conducting cost-effectiveness analyses.


Comparison of kinesiophobia in patients with rheumatoid arthritis and systemic lupus erythematosus.
Patients with rheumatic disease often experience arthritis and chronic joint pain, which can lead them to avoid movement, known as kinesiophobia. This may result in decreased mobility and endurance, as well as social isolation and depression. This study aimed to assess and compare the prevalence of kinesiophobia among patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), and its association with disease activity, functional status, fear of falling, and fatigue. A cross-sectional study was conducted with 124 RA, 76 SLE patients, and 87 healthy controls. The tampa kinesiophobia scale (TKS) was used to assess kinesiophobia. The disease activity was assessed using Disease Activity Score 28 (DAS28) in RA, and systemic lupus erythematosus Disease Activity Index 2000 (SLEDAI-2K) in SLE patients. The Health Assessment Questionnaire (HAQ) was used to assess functional status, the Falls Efficacy Scale International (FES-I) was used to assess fear of falling, and the Fatigue Severity Scale (FSS) was used to assess fatigue levels. Kinesiophobia was significantly more prevalent in RA patients compared to SLE patients [77.4% vs. 63.2%, odds ratio (OR): 2, 95% CI: 1.07-3.75; p<0.05]. The mean TKS score was 41.42±6.95 in RA patients, and 37.84±8.85 in SLE (p=0.005). TKS scores were positively correlated with DAS28 in RA patients; however, no correlation was found between SLEDAI-2K in SLE patients. A positive correlation was observed between TKS scores and the HAQ, FSS, and FES-I, both in RA and SLE patients. RA patients exhibited significantly higher kinesiophobia scores compared to SLE patients. The higher rate of kinesiophobia levels in RA patients compared to SLE patients suggests an association between kinesiophobia and the more destructive course of RA, indicating that tight control of the disease is important. Kinesiophobia was associated with functional impairment, fear of falling, and fatigue. Timely identification and intervention for patients with kinesiophobia are essential to prevent progression, mitigate long-term consequences, and maintain functional capacity.


Cytomegalovirus Reactivation in Patients with Sepsis in an Intensive Care Unit in Portugal.
In the last few years, cytomegalovirus reactivation has been considered an aggravating factor for septic patients in Intensive Care units. The main objectives of this study were to determine cytomegalovirus reactivation in patients with a diagnosis of sepsis admitted to an intensive care unit, and whether this reactivation was related to the evolution of the patient's clinical condition. The detection of cytomegalovirus DNA was performed by real-time polymerase chain reaction and the concentration of nine cytokines (IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL- TNF-α, and INFγ) were determined by a Multiplex ELISA technique. Eight of 22 septic patients (36.3%) from the Intensive Care Unit of the Hospital da Luz had cytomegalovirus reactivation. No association was found between cytomegalovirus reactivation and gender, age, length of Intensive Care unit stay, duration of mechanical ventilation, and patient death. No significant differences were found in cytokine concentrations in patients with and without reactivation. However, patients with cytomegalovirus reactivation had a longer hospital stay from Intensive Care unit entry to hospital discharge or patient death (p = 0.025). Despite the low sampling rate, the present study suggests that reactivation is a frequent event in patients diagnosed with sepsis and may be related to prolonged hospital stay in these patients. The overall analysis of the results obtained and the literature review do not support the concept that cytomegalovirus monitoring should be implemented in routine practice, but it seems prudent to wait for further randomized trials using antiviral prophylaxis, before assuming a definitive attitude towards the role of cytomegalovirus in sepsis.


Glucocorticoid receptor-regulated TcLEC2 expression triggers somatic embryogenesis in Theobroma cacao leaf tissue.
Theobroma cacao, the source of cocoa, is a crop of particular importance in many developing countries. Availability of elite planting material is a limiting factor for increasing productivity of Theobroma cacao; therefore, the development of new strategies for clonal propagation is essential to improve farmers' incomes and to meet increasing global demand for cocoa. To develop a more efficient embryogenesis system for cacao, tissue was transformed with a transgene encoding a fusion of Leafy Cotyledon 2 (TcLEC2) to a glucocorticoid receptor domain (GR) to control nuclear localization of the protein. Upon application of the glucocorticoid dexamethasone (dex), downstream targets of LEC2 involved in seed-development were up-regulated and somatic embryos (SEs) were successfully regenerated from TcLEC2-GR transgenic flower and leaf tissue in large numbers. Immature SEs regenerated from TcLEC2-GR leaves were smaller in size than immature SEs from floral tissue, suggesting a different ontogenetic origin. Additionally, exposure of TcLEC2-GR floral explants to dex increased the number of SEs compared to floral explants from control, non-transgenic trees or from TcLEC2-GR floral explants not treated with dex. Testing different durations of exposure to dex indicated that a three-day treatment produced optimal embryo regeneration. Leaf derived SEs were successfully grown to maturity, converted into plants, and established in the greenhouse, demonstrating that these embryos are fully developmentally competent. In summary, we demonstrate that regulating TcLEC2 activity offers a powerful new strategy for optimizing somatic embryogenesis pipelines for cacao.


[Oxyproline excretion in the urine in frostbite].
Hyperoxyprolinuria associated with the processes occuring in frostbitten tissues is observed in frostbites of II--III degree. In frostbites of I--II and III--IV degree the level of urine oxyproline excretion does not differ from the normal one throughout the posttraumatic period. In frostbites of I--II degree this is due to a limited volume of tissue injury, in frostbites of III--IV degree-to the fixation of decay products in the pathological focus, resulting from blood circulation disorders in the frostbitten zone. Differences in oxyprolinuria level may serve as a useful tool for differential diagnosis of frostbites.


Interfacility transfer and mortality for patients with ruptured abdominal aortic aneurysm.
Patients receiving interfacility transfer to a higher level of medical care for ruptured abdominal aortic aneurysms (rAAAs) are an important minority that are not well characterized and are typically omitted from outcomes and quality indicator studies. Our objective was to compare patients transferred for treatment of rAAAs with those treated without transfer, with particular emphasis on mortality and resource utilization. We linked longitudinal data from 2005 to 2010 Healthcare Cost and Utilization Project (HCUP) State Inpatient Databases and Emergency Department Databases from California, Florida, and New York. Patients were identified using International Classification of Diseases-Ninth Revision-Clinical Modification codes. Our main outcome variables were mortality, length of stay, and cost. Data included discharge information on the transfer-out and transfer-in hospital. We used univariate and multivariate analysis to identify variables independently associated with transfer and in-hospital mortality. Of 4439 rAAA patients identified with intent to treat, 847 (19.1%) were transferred before receiving operative repair. Of those transferred, 141 (17%) died without undergoing AAA repair. By multivariate analysis, increasing age in years (odds ratio [OR] 0.98; 95% confidence interval [CI], 0.97-0.99; P < .001), private insurance vs Medicare (OR, 0.62; 95% CI, 0.47-0.80; P < .001), and increasing comorbidities as measured by the Elixhauser Comorbidity Index (OR, 0.90; 95% CI, 0.86-0.95; P < .001) were negatively associated with transfer. Weekend presentation (OR, 1.23; 95% CI, 1.02-1.47; P = .03) was positively associated with transfer. Transfer was associated with a lower operative mortality (adjusted OR, 0.81; 95% CI, 0.68-0.97; P < .02) but an increased overall mortality when including transferred patients who died without surgery (OR, 1.30; 95% CI, 1.05-1.60; P = .01). Among the transferred patients, there was no significant difference in travel distance between those who survived and those who died (median, 28.7 vs 25.8 miles; P = .07). Length of stay (median, 10 vs 9 days; P = .008), and hospital costs ($161,000 vs $146,000; P = .02) were higher for those transferred. The survival advantage for patients transferred who received treatment was eclipsed by increased mortality of the transfer process. Including 17% of transferred patients who died without receiving definitive repair, mortality was increased for patients transferred for rAAA repair compared with those not transferred after adjusting for demographic, clinical, and hospital factors. Transferred patients used significantly more hospital resources. Improving systems and guidelines for interfacility transfer may further improve the outcomes for these patients and decrease associated hospital resource utilization.


Tamoxifen-induced enhancement of calcium signaling in glioma and MCF-7 breast cancer cells.
The antiestrogen tamoxifen is commonly used to treat breast cancer, but it also has therapeutic activity in several other types of cancer. Many of these tumors, including malignant gliomas, are estrogen receptor negative. Nonetheless, high concentrations of tamoxifen can directly reduce cell proliferation in some of these tumors and induce apoptosis. In this study, the role of tamoxifen in calcium signaling and calcium-induced cell death was studied in both malignant glioma cell lines and MCF-7 breast cancer cells. Tamoxifen potently increased the spatial expansion of calcium waves by 30-150% while significantly enhancing and prolonging agonist-induced calcium elevations. Furthermore, tamoxifen pretreatment accelerated calcium ionophore-induced death by more than 20 min, suggesting that tamoxifen lowered cellular resistance to calcium loads. In contrast to its potentiating of calcium signaling in tumors, tamoxifen had no significant effect on calcium signaling in cultures of primary astrocytes from either human or rat brain. This study demonstrates the existence of calcium signaling in breast cancer and glioma cells and identifies tamoxifen as a potential modulator of tumor-associated calcium signaling.


Fluorescent functionalised naphthalimides and their Au(i)-NHC complexes for potential use in cellular bioimaging.
A series of cationic, dihydroimidazolinium-functionalized 1,8-naphthalimide fluorophores have been isolated as their hexafluorophosphate salts, [HL]PF6. These pro-ligands react with [AuCl(tht)] in the presence of base to form N-heterocyclic carbene (NHC) complexes, [AuCl(L)]. Two X-ray structures represent a pro-ligand and complex pairing: the latter reveals the two-coordinate linear geometry of the NHC-Au(i) species, as well as intermolecular interactions supported by both ligand π-π stacking and a weak aurophilic interaction of 3.3205(6) Å. The luminescence properties of the pro-ligands and complexes are dominated by the ICT character of the substituted fluorophore at ca. 500 nm, which is further modulated via functionalization at the 4-position of the naphthalimide. Cytotoxicity assessments were performed for all [HL]PF6 and [AuCl(L)] species against LOVO, MCF-7, A549 and PC3 cell lines; added lipophilicity seems to correlate with increased cytotoxicity. Confocal fluorescence microscopy was undertaken on a selected [HL]PF6 and [AuCl(L)] species and showed that the intracellular distribution is dependent upon the specific ligand structure. More detailed co-localisation studies show that selected examples present a predominant lysosomal staining pattern. FLIM studies exemplified the applicability of these probes, and secondly suggest that fluorescence lifetime could be used to provide information on the integrity of the complex and thus liberation of gold in a biological environment.


Two-year-olds use adults' but not peers' points.
In the current study, 24- to 27-month-old children (N = 37) used pointing gestures in a cooperative object choice task with either peer or adult partners. When indicating the location of a hidden toy, children pointed equally accurately for adult and peer partners but more often for adult partners. When choosing from one of three hiding places, children used adults' pointing to find a hidden toy significantly more often than they used peers'. In interaction with peers, children's choice behavior was at chance level. These results suggest that toddlers ascribe informative value to adults' but not peers' pointing gestures, and highlight the role of children's social expectations in their communicative development.


Functional specialization within a vesicle tethering complex: bypass of a subset of exocyst deletion mutants by Sec1p or Sec4p.
The exocyst is an octameric protein complex required to tether secretory vesicles to exocytic sites and to retain ER tubules at the apical tip of budded cells. Unlike the other five exocyst genes, SEC3, SEC5, and EXO70 are not essential for growth or secretion when either the upstream activator rab, Sec4p, or the downstream SNARE-binding component, Sec1p, are overproduced. Analysis of the suppressed sec3Delta, sec5Delta, and exo70Delta strains demonstrates that the corresponding proteins confer differential effects on vesicle targeting and ER inheritance. Sec3p and Sec5p are more critical than Exo70p for ER inheritance. Although nonessential under these conditions, Sec3p, Sec5p, and Exo70p are still important for tethering, as in their absence the exocyst is only partially assembled. Sec1p overproduction results in increased SNARE complex levels, indicating a role in assembly or stabilization of SNARE complexes. Furthermore, a fraction of Sec1p can be coprecipitated with the exoycst. Our results suggest that Sec1p couples exocyst-mediated vesicle tethering with SNARE-mediated docking and fusion.


Pregnancy after breast reconstruction with a transverse rectus abdominis musculocutaneous flap.
The transverse rectus abdominis musculocutaneous flap has been used for breast reconstruction with increasing frequency in recent years. It has not previously been reported whether or not an abdominal wall weakened by the transfer of this flap can withstand the added stress of pregnancy. An experience with one such patient who successfully carried a pregnancy to term is described.


Monitoring Psychometric States of Recovery to Improve Performance in Soccer Players: A Brief Review.
In order to maximize adaptations and to avoid nonfunctional overreaching syndrome or noncontact injury, coaches in high-performance sports must regularly monitor recovery before and after competitions/important training sessions and maintain well-being status. Therefore, quantifying and evaluating psychometric states of recovery during the season in sports teams such as soccer is important. Over the last years, there has been substantial growth in research related to psychometric states of recovery in soccer. The increase in research on this topic is coincident with the increase in popularity obtained by subjective monitoring of the pre-fatigue state of the players before each training sessions or match with a strong emphasis on the effects of well-being or recovery state. Among the subjective methods for players' control, the Hooper index (HI) assesses the quality of sleep during the previous night, overall stress, fatigue, and delayed-onset muscle soreness. Additionally, the total quality of recovery (TQR) scale measures recovery status. The HI and TQR recorded before each training session or match were affected by the variability of training load (TL) and influenced the physical and technical performances, and the affective aspects of soccer players. Researchers have recommended wellness monitoring soccer players' psychometric state of recovery before each training session or match in order to detect early signs of fatigue and optimize high-level training performance. This method allows for better detecting signs of individual fatigue and allows coaches to adapt and readjust the TL, and avoid physical and technical gaps in order to improve the performance of soccer players.


Specialist training: workplace-based assessments impact on teaching, learning and feedback to support competency-based postgraduate programs.
Workplace-based assessments (WBAs) are part of a competency-based curriculum where training progression is dependent on the achievement of defined competencies in a real-world clinical environment. There is a significant literature gap on the impact of WBAs implemented in resource constrained countries and their contextual challenges. This study aimed to examine the use, impact, and educational context of WBAs in South African medical specialist training programs drawing on perspectives from both trainees and trainers to identify educational challenges and propose effective solutions. A mixed methods national electronic survey was conducted with specialist medical trainees and supervising trainers from all eight specialist training institutions in South Africa involving 16 specialities. The survey responses were voluntary and anonymous. The survey was closed after seven months when data saturation was achieved. Descriptive statistical analysis was performed using SPSS Version 27 (SPSS Inc, 2012, Chicago, IL) for the quantitative analysis. The thematic coding framework for the qualitative analysis was facilitated by NVivo Version 12 software. There were 108 ethnically diverse supervising trainers and 248 specialist trainees' survey respondents. Across the 16 medical specialities, 45% of the respondents were using WBAs. Despite contextual resource and staff challenges, this study found that WBAs had a positive impact to Kirkpatrick level 2 in providing actionable feedback to improve competency. WBA users had a significantly higher rating for trainee supervision (p < 0.01), general quality of feedback on trainee competence (< 0.01) and the specialist training program (p = 0.03) compared to WBA non-users. They also had a higher rating for the assessment of the trainee as a professional (p < 0.01); scholar (p < 0.01); communicator (p < 0.01); collaborator (p = 0.001) and leader/manager (p < 0.001) based on the AfriMEDS competency framework. Racism, sexism and favouritism were challenges that negatively affected the training programs. Overall, this study reports that the use of WBAs had a substantially favourable impact on teaching, learning, feedback and supports a competency-based approach to specialist training programs. Addressing the contextual concerns that negatively impact training; training the trainees and trainers about their relationship, roles and responsibilities; and focusing on a trainee-centred, inclusive and empowering teaching approach will help further enhance its effectiveness.


Churg-Strauss Syndrome Following Vaccination Against 2010 Influenza A (H1N1): A Case Report.
Churg-Strauss syndrome (CSS) is a systemic inflammatory disorder characterized by asthma, transient pulmonary infiltration, hyper-eosinophilia, and systemic vasculitis. Reported triggering factors include infections, drugs, allergic desensitization, and vaccinations, although cases involving the latter two are extremely rare. Herein, we describe a patient who developed CSS after receiving an H1N1 vaccination. A 55-year-old woman presented with fever, skin eruptions, and sensory impairment of her feet within one week after an H1N1 vaccine injection. A chest X-ray showed pulmonary infiltrations in both lower lung fields. Eosinophilia was noted in a hematological test, and an electrophysiological study revealed a pattern of mononeuritis multiplex. A skin biopsy was performed which revealed palisading necrotizing granuloma around a degenerated dermis and eosinophilic infiltration of the blood vessel walls. These findings combined with the hematological and electrophysiological findings met the criteria of CSS according to the American College of Rheumatology. The patient recovered well after steroid treatment. This case highlights the possibility that the H1N1 vaccination can trigger CSS. Due to the rarity of reported autoimmune events after vaccine administration and the obscure causal association between autoimmunity and a vaccine, further post-marketing surveillance and research are necessary to clarify the relationship and identify risk factors.


Nanoencapsulation strategies for the delivery of novel bifunctional antioxidant/σ1 selective ligands.
Nowadays sigma-1 receptors are considered as new therapeutic objectives for central nervous system neurodegenerative diseases. Among different molecules, alpha lipoic acid has been identified as a natural potent antioxidant drug, whose therapeutic efficacy is limited by its many drawbacks, such as fast metabolism, poor bioavailability and high physico-chemical instability. Alfa-lipoic acid derivatives have been recently developed demonstrating their neuroprotective activity and effectiveness in different types of oxidative stress. In this work, two derivatives containing an amide or an ester functional group with different lipophilicity, were selected for their important affinity for sigma-1 receptors. Herein, in order to improve the in vitro stability and antioxidant effectiveness of alpha-lipoic acid derivatives, we focused our efforts in the nanoencapsulation strategies. Aqueous-core nanocapsules for the delivery of the hydrophilic compound and nanostructured lipid carrier for the lipophilic derivative, were properly designed and prepared using a direct or inverse eco-friendly organic solvent-free procedure. All nanosystems were characterized in terms of mean size, polydispersity, stability, morphology, encapsulation efficiency and in vitro release profiles. In order to evaluate the nanocarriers biocompatibility and antioxidant effectiveness, in vitro biological studies (cell viability, total antioxidant capacity and total oxidative status) were developed on primary human whole blood cell cultures, on both unloaded and derivatives-loaded nanodevices.


Alpha-Lipoic Acid as an Antioxidant Strategy for Managing Neuropathic Pain.
Neuropathic pain (NP) is the most prevalent and debilitating form of chronic pain, caused by injuries or diseases of the somatosensory system. Since current first-line treatments only provide poor symptomatic relief, the search for new therapeutic strategies for managing NP is an active field of investigation. Multiple mechanisms contribute to the genesis and maintenance of NP, including damage caused by oxidative stress. The naturally occurring antioxidant alpha-lipoic acid (ALA) is a promising therapeutic agent for the management of NP. Several pre-clinical in vitro and in vivo studies as well as clinical trials demonstrate the analgesic potential of ALA in the management of NP. The beneficial biological activities of ALA are reflected in the various patents for the development of ALA-based innovative products. This review demonstrates the therapeutic potential of ALA in the management of NP by discussing its analgesic effects by multiple antioxidant mechanisms as well as the use of patented ALA-based products and how technological approaches have been applied to enhance ALA's pharmacological properties.


Air in the kidney: between emphysematous pyelitis and pyelonephritis.
Presence of air in the kidney can be problematic as the location of the air in different parts of the kidney greatly affects the subsequent management and outcome of the patient. We present here a case of a patient who had emphysematous pyelitis, in which CT scan was able to display presence of air only in the collecting system, thus differentiating this condition from the more fulminant emphysematous pyelonephritis. This leads to a more favourable prognosis and outcome to the patient.


Use of immunoglobulin m cross-reactions in differential diagnosis of human flaviviral encephalitis infections in the United States.
To define the virus specificity of the immunoglobulin M (IgM) antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA) among the medically important members of the Japanese encephalitis (JE) virus serocomplex of flaviviruses, 103 IgM-positive human serum samples from patients with confirmed West Nile (WN) virus, St. Louis encephalitis (SLE) virus, or JE virus infections were assembled and simultaneously tested against all three viral antigens in a standardized MAC-ELISA. Of the serum samples tested, 96 (93%) showed higher positive-to-negative absorbance ratios (P/Ns) with the infecting virus antigen compared to those obtained with the other two virus antigens. Of the seven specimens with higher P/Ns with heterologous virus antigens, six were from patients with SLE virus infections (the serum samples had higher levels of reactivity with WN virus antigen) and one was from a patient with a JE virus infection (this serum sample also had a higher level of reactivity with WN virus antigen). Not surprisingly, similar virus specificity was observed with WN virus-elicited IgM in cerebrospinal fluid. As shown in previous studies, a subset of these specimens was even less reactive in the MAC-ELISA with dengue virus, a member of a different flavivirus serocomplex. The degree of virus cross-reactivity did not appear to be related to days postonset, at least during the first 40 days of infection. Infections with WN virus could be correctly distinguished from infections with SLE virus on the basis of the observed anti-viral IgM cross-reactivities alone 92% of the time. Infections with SLE virus resulted in antibody that was more cross-reactive, so identification of SLE virus as the infecting agent by use of MAC-ELISA cross-reactivity alone was more problematic.


Common Data Elements for Acute Coronary Syndrome: Analysis Based on the Unified Medical Language System.
Standardization in clinical documentation can increase efficiency and can save time and resources. The objectives of this work are to compare documentation forms for acute coronary syndrome (ACS), check for standardization, and generate a list of the most common data elements using semantic form annotation with the Unified Medical Language System (UMLS). Forms from registries, studies, risk scores, quality assurance, official guidelines, and routine documentation from four hospitals in Germany were semantically annotated using UMLS. This allowed for automatic comparison of concept frequencies and the generation of a list of the most common concepts. A total of 3710 forms items from 86 sources were semantically annotated using 842 unique UMLS concepts. Half of all medical concept occurrences were covered by 60 unique concepts, which suggests the existence of a core dataset of relevant concepts. Overlap percentages between forms were relatively low, hinting at inconsistent documentation structures and lack of standardization. This analysis shows a lack of standardized and semantically enriched documentation for patients with ACS. Efforts made by official institutions like the European Society for Cardiology have not yet been fully implemented. Utilizing a standardized and annotated core dataset of the most important data concepts could make export and automatic reuse of data easier. The generated list of common data elements is an exemplary implementation suggestion of the concepts to use in a standardized approach.


Tobacco as a risk factor in bladder cancer.
A case-control study concerning smoking habits has been performed to determine risk factors for bladder cancer. The population attributable risk (AP) for cigarettes, cigars/cigarillos, chewing tobacco and pipe yielded a statistical explanation of 33%, 23%, 9% and 17%, respectively. A logistic regression analysis revealed cigarette smoking as the most pronounced determinant with an independent effect beyond the other smoking variables corresponding to a relative risk (RR) of 2.15. The life-time consumption of all kinds of tobacco showed a positive dose-response relationship to risk for bladder cancer.


Genomic characterization of novel circular ssDNA viruses from insectivorous bats in Southern Brazil.
Circoviruses are highly prevalent porcine and avian pathogens. In recent years, novel circular ssDNA genomes have recently been detected in a variety of fecal and environmental samples using deep sequencing approaches. In this study the identification of genomes of novel circoviruses and cycloviruses in feces of insectivorous bats is reported. Pan-reactive primers were used targeting the conserved rep region of circoviruses and cycloviruses to screen DNA bat fecal samples. Using this approach, partial rep sequences were detected which formed five phylogenetic groups distributed among the Circovirus and the recently proposed Cyclovirus genera of the Circoviridae. Further analysis using inverse PCR and Sanger sequencing led to the characterization of four new putative members of the family Circoviridae with genome size ranging from 1,608 to 1,790 nt, two inversely arranged ORFs, and canonical nonamer sequences atop a stem loop.


Influence of the liver on the profile of circulating antigens recognized by antiserum against hepatoma membrane glycoproteins.
Rat hepatoma tissue culture cells have retained only a few liver-specific membrane functions. To find out whether transformation of these cells led to the appearance of tumor-specific membrane glycoprotein structures, which might also be present in released form in the circulation of tumor-bearing animals, we compared immunologically the sera and ascites fluids of normal and of hepatoma-bearing rats. For structural identification, we used polyspecific antibodies against the membrane glycoproteins of the hepatoma cells. Crossed immunoelectrophoresis of the body fluids resulted in the separation of several antigens, one of which appears to be tumor specific. Immunoelectrophoretic analyses of the glycoproteins from the membranes of liver and hepatoma cells revealed many shared antigens; in one instance, we could assign the membrane associated form to major antigen detected in the serum. Characterization of the molecular size and charge of the two selected antigens by two-dimensional gel electrophoresis indicated that the hepatoma cells products were not completely sialylated. In the case of the circulating, hepatoma-specific antigen, this property appears to result in hepatic clearance of the more basic ("asialo") portion of the glycoprotein species via the receptor-mediated uptake of galactose-terminated glycoproteins. The result was the accumulation of the acidic forms of the antigen in the serum and ascites fluid of tumor-bearing rats.


Overcoming barriers to recognizing and reporting child abuse.
Infants and children under age 3 years have the highest risk of dying from child abuse and neglect. Clinicians treating children must recognize and report child abuse. Barriers to consistent recognition and reporting leave children in harm's way. Often, the signs of abuse in very young children are subtle, and clinicians may fail to recognize and report these signs. Clinicians also must understand the role of bias in the reporting of child abuse and ways to address abuse individually and as part of a larger system.


Lipoprotein-associated phospholipase A2 and risk of dementia in the Cardiovascular Health Study.
To evaluate associations between Lipoprotein-associated phospholipase A2 (Lp-PLA2) mass and activity with risk of dementia and its subtypes. Analysis were completed on 3320 participants of the Cardiovascular Health Study (CHS), a population-based longitudinal study of community-dwelling adults age ≥65 years followed for an average of 5.4 years. Baseline serum Lp-PLA2 mass was measured using a sandwich enzyme immunoassay and Lp-PLA2 activity utilized a tritiated-platelet activating factor activity assay. Cox proportional hazards regression assessed the relative risk of incident dementia with higher baseline Lp-PLA2 adjusting for demographics, cardiovascular disease (CVD) and risk factors, inflammation markers and apolipoprotein E (APOE) genotype. Each standard deviation higher Lp-PLA2 mass and activity were related to increased risk of dementia (fully adjusted HR: 1.11 per SD, 95% CI: 1.00-1.24 for mass; HR: 1.12 per SD, 95% CI: 1.00-1.26 for activity). Persons in the highest quartile of Lp-PLA2 mass were 50% more likely to develop dementia than those in the lowest quartile in adjusted models (HR: 1.49; 95% CI: 1.08-2.06). Among dementia subtypes, the risk of AD was increased two-fold in the highest compared to lowest quartile of Lp-PLA2 mass (adjusted HR: 1.98, 95% CI: 1.22-3.21). Results were attenuated in models of mixed dementia and VaD. Lp-PLA2 activity also doubled the risk of mixed dementia in the highest compared to lowest quartile (HR: 2.21, 95% CI: 1.12-4.373). These data support Lp-PLA2 as a risk factor for dementia independent of CVD and its risk factors. Further study is required to clarify the role of Lp-PLA2-related mechanisms in dementia subtypes.


Ultrasonography in the pathway to an optimal standard of care of hidradenitis suppurativa: the Italian Ultrasound Working Group experience.
Ultrasound (US) is a real-time non-invasive technique that has been demonstrated to support an early diagnosis and a more precise assessment of hidradenitis suppurativa (HS). To compare the clinical and US evaluation of a series of HS patients. 434 HS patients (259 F, 175 M; mean age 33.82 ±13.31 years) observed across 19 Italian dermatology centres [members of the Italian Ultrasound Working Group (IUWG)] were enrolled in a retrospective study. Clinical staging was obtained by the Hidradenitis Suppurativa Physician's Global Assessment score (HS-PGA), while the ultrasonographic staging was determined by the US HS-PGA, based on the same scores as clinical HS-PGA but performed with the aid of US. At the end of the study, the mean clinical and US HS-PGA scores were 2.70 and 2.92, respectively. Direct comparison of clinical and ultrasonographic assessment revealed that a higher proportion of patients was classified as having moderate and very severe disease by US. In particular, 117 patients (26.96%) had a worse classification by US HS-PGA compared to clinical assessment. Our findings confirm that the use of clinical grading only to assess HS severity may underestimate the real disease severity. US examination can be considered an essential non-invasive imaging tool available to dermatologists for a more accurate diagnosis, staging, treatment planning and monitoring of HS and should be included in the pathway to an optimal standard of care of HS.


Requirement for Xvent-1 and Xvent-2 gene function in dorsoventral patterning of Xenopus mesoderm.
Xvent-1 and Xvent-2 are members of a novel homeobox subfamily that have been implicated in dorsoventral patterning in Xenopus mesoderm and are thought to function in BMP signalling. Here we investigate the requirement for Xvent function by employing two dominant-negative strategies. Loss of Xvent function dorsalizes ventral mesoderm, induces secondary embryonic axes and directly neuralizes ectoderm. We further find that (1) Xvents act as transcriptional repressors, (2) Xvents function in an additive fashion and (3) a surprising number of genes are able to rescue dominant-negative Xvent phenotypes including Bmp-4, Smad-1 and wild-type Xvents and Xhox3, but not Xwnt-8. The results show that Xvent-1 and Xvent-2 are essential for ventral mesoderm formation and for preventing neural differentiation. A model is suggested to explain how Bmp-4 positional information is converted into distinct cellular responses.


Current Practice of Therapeutic Drug Monitoring of Biopharmaceuticals in Psoriasis Patients.
The high prevalence of psoriasis and the high spending on pharmaceuticals motivate a more evidence-based and cost-effective usage of biopharmaceuticals. A growing body of evidence exists that the implementation of therapeutic drug monitoring for biopharmaceuticals in psoriasis patients optimizes patient management and clinical outcome and enhances their efficacy. Therefore, the aim of this review was to give an overview of the literature on therapeutic drug monitoring of biopharmaceuticals in the treatment of psoriasis and to provide the useful information to dermatologists to improve health care in psoriasis patients.


Evaluation of a newly developed down-flow immunoassay for detection of serum mannan antigens in patients with candidaemia.
A down-flow immunoassay has been developed to detect serum mannan antigens, and the test was recently marketed as the Unimedi Candida monotest. Using 251 serum samples from 105 patients with candidaemia, a comparison of the Unimedi Candida monotest with the Cand-Tec latex agglutination test and 2 microplate enzyme immunoassay tests (Platelia Candida Ag test and Unimedi Candida) was conducted. One hundred and seventy-five febrile patients without clinical and microbiological evidence of fungal infections and pneumocytosis were examined as controls. The Cand-Tec test had a sensitivity of 38% and a specificity of 82%. The sensitivity and specificity of the Platelia Candida Ag test, the Unimedi Candida and the Unimedi Candida monotest were 53 and 92%, 69 and 89% and 82 and 96%, respectively. The sensitivity of the Unimedi Candida monotest was significantly (P<0.01) higher than that of the Plateria Candida Ag test for diagnosing candidaemia caused by Candida parapsilosis. The beta-D-glucan assay had a high sensitivity of 95%, with a specificity of 84%. Of 74 patients with candidaemia whose sera were available before or on positive blood culture sampling, 29 (39%), 38 (51%) and 48 (65%) patients had antigenemia detected using the Platelia Candida Ag test, the Unimedi Candida and the Unimedi Candida monotest, respectively. The Unimedi Candida monotest seems to be a promising tool for the early diagnosis of invasive candidiasis, because the test was sensitive, simple, rapid (approx. 1 h) and cost-effective.


Investigating Choice Experiments for Preferences of Older People (ICEPOP): evaluative spaces in health economics.
This paper deals with three concerns about the evaluative framework that is currently dominant within health economics. These concerns are: that the evaluative framework is concerned entirely with health; that the evaluative framework has an individualistic focus on patients alone; and that the methods used to estimate 'health' within the current evaluative framework could be improved both in terms of the generation of descriptive systems and in using valuation methods that rely less on people's ability to express their preferences on a cardinal scale. In exploring these issues the Investigating Choice Experiments for Preferences of Older People (ICEPOP) programme has explicitly focused on both the topic of older people and the methods of discrete choice experiments. A capability index has been developed and attributes for an economic measure of end-of-life care are currently being generated, providing the possibility of extending the evaluative framework beyond health alone. A measure of carer's experience and a framework for extending measurement in end-of-life care to loved ones are both also in development, thus extending the evaluative framework beyond the patient alone. Rigorous qualitative methods employing an iterative approach have been developed for use in constructing attributes, and best-worst scaling has been utilized to reduce task complexity and provide insights into heterogeneity. There are a number of avenues for further research in all these areas, but in particular there is need for greater attention to be paid to the theory underlying the evaluative framework within health economics.