An alternative phase II/III design for continuous endpoints.
The success rate of drug development has been declined dramatically in recent years and the current paradigm of drug development is no longer functioning. It requires a major undertaking on breakthrough strategies and methodology for designs to minimize sample sizes and to shorten duration of the development. We propose an alternative phase II/III design based on continuous efficacy endpoints, which consists of two stages: a selection stage and a confirmation stage. For the selection stage, a randomized parallel design with several doses with a placebo group is employed for selection of doses. After the best dose is chosen, the patients of the selected dose group and placebo group continue to enter the confirmation stage. New patients will also be recruited and randomized to receive the selected dose or placebo group. The final analysis is performed with the cumulative data of patients from both stages. With the pre-specified probabilities of rejecting the drug at each stage, sample sizes and critical values for both stages can be determined. As it is a single trial with controlling overall type I and II error rates, the proposed phase II/III adaptive design may not only reduce the sample size but also improve the success rate. An example illustrates the applications of the proposed phase II/III adaptive design.


3'-Intercalation of a N2-dG 1R-trans-anti-benzo[c]phenanthrene DNA adduct in an iterated (CG)3 repeat.
The conformation of the 1 R,2 S,3 R,4 S-benzo[ c]phenanthrene- N (2)-dG adduct, arising from trans opening of the (+)-1 S,2 R,3 R,4 S- anti-benzo[ c]phenanthrene diol epoxide, was examined in 5'- d(ATCGC XCGGCATG)-3'.5'-d(CATGCCG CGCGAT)-3', where X = 1 R,2 S,3 R,4 S-B[ c]P- N (2)-dG. This duplex, derived from the hisD3052 frameshift tester strain of Salmonella typhimurium, contains a (CG) 3 iterated repeat, a hotspot for frameshift mutagenesis. NMR experiments showed a disconnection in sequential NOE connectivity between X (4) and C (5), and in the complementary strand, they showed another disconnection between G (18) and C (19). In the imino region of the (1)H NMR spectrum, a resonance was observed at the adducted base pair X (4) x C (19). The X (4) N1H and G (18) N1H resonances shifted upfield as compared to the other guanine imino proton resonances. NOEs were observed between X (4) N1H and C (19) N (4)H and between C (5) N (4)H and G (18) N1H, indicating that base pairs X (4) x C (19) and C (5) x G (18) maintained Watson-Crick hydrogen bonding. No NOE connectivity was observed between X (4) and G (18) in the imino region of the spectrum. Chemical shift perturbations of greater than 0.1 ppm were localized at nucleotides X (4) and C (5) in the modified strand and G (18) and C (19) in the complementary strand. A total of 13 NOEs between the protons of the 1 R-B[ c]Ph moiety and the DNA were observed between B[ c]Ph and major groove aromatic or amine protons at base pairs X (4) x C (19) and 3'-neighbor C (5) x G (18). Structural refinement was achieved using molecular dynamics calculations restrained by interproton distances and torsion angle restraints obtained from NMR data. The B[ c]Ph moiety intercalated on the 3'-face of the X (4) x C (19) base pair such that the terminal ring of 1 R-B[ c]Ph threaded the duplex and faced into the major groove. The torsion angle alpha' [X (4)]-N3-C2-N2-B[ c]Ph]-C1 was calculated to be -177 degrees, maintaining an orientation in which the X (4) exocyclic amine remained in plane with the purine. The torsion angle beta' [X (4)]-C2-N2-[B[ c]Ph]-C1-C2 was calculated to be 75 degrees. This value governed the 3'-orientation of the B[ c]Ph moiety with respect to X (4). The helical rise between base pairs X (4) x C (19) and C (5) x G (18) increased and resulted in unwinding of the right-handed helix. The aromatic rings of the B[ c]Ph moiety were below the Watson-Crick hydrogen-bonding face of the modified base pair X (4) x C (19). The B[c]Ph moiety was stacked above nucleotide G (18), in the complementary strand.


Conservative treatment of patients with thromboangiitis obliterans or cannabis-associated arteritis presenting with critical lower limb ischaemia.
Proper management of patients with thromboangiitis obliterans (TAO) or cannabis-associated arteritis (CAA), presenting with critical lower limb ischaemia (CLI) remains controversial, and data are limited. Patients with TAO or CAA presenting with CLI between 2011 and 2016 were retrospectively evaluated. Patients requiring primary intervention were excluded. Conservative treatment included: (a) weight-adjusted bemiparin plus six hours/day intravenous iloprost for 28 days, (b) aspirin (100 mg/day) plus cilostazol (100 mg twice/day) after discharge, and (c) strict recommendations/monitoring for smoking cessation. Main outcomes included symptom recession, ankle-brachial index (ABI) improvement, and healing of lesions at the time of discharge as well as amputation, revascularization, and abstinence rate during follow-up. Overall, 23 patients (TAO: 15; CAA: 8) were included within six years, none of the patients reported any other factor than smoking. All patients presented with rest pain and 12 patients with ulcer or necrotic lesions. Mean ABI measurement at presentation was 0.46 ± 0.2, after 28 days of treatment, all patients showed improvement regarding clinical picture and ABI measurement (0.54 ± 0.1; p < 0.05). During follow-up, only three patients underwent bypass surgery and two patients underwent major amputation, although the smoking abstinence rate was very low (13 %). Intravenous iloprost plus bemiparin for 28 days together with per os aspirin plus cilostazol seem to produce promising results in patients with TAO/CAA, treated for CLI, even with a low smoking abstinence rate. However, larger series are needed to further evaluate inter-group differences and potential prognostic factors.


Cutaneous nocardiosis in a cat.
A 7-year-old cat was found to have multiple cutaneous lesions caused by Nocardia asteroides infection. The cat was treated successfully with potentiated sulfonamides and a combination of ampicillin and erythromycin.


Abrupt category shifts during real-time person perception.
Previous studies have suggested that real-time person perception relies on continuous competition, in which partially active categories smoothly compete over time. Here, two studies demonstrated the involvement of a different kind of competition. In Study 1, before participants selected the correct sex category for morphed faces, their mouse trajectories often exhibited a continuous attraction toward the incorrect category that increased with sex-category ambiguity, indicating continuous competition. On other trials, however, trajectories initially pursued the incorrect category and then abruptly redirected toward the correct category, suggesting early incorrect category activation that was rapidly reversed later in processing. These abrupt category reversals also increased with ambiguity. In Study 2, participants were presented with faces containing a sex-typical or sex-atypical hair cue, in a context in which the norm was either sex-typical targets (normative context) or sex-atypical targets (counternormative context). Sex-atypical targets induced greater competition in the normative context, but sex-typical targets induced greater competition in the counternormative context. Together, these results demonstrate that categorizing others involves both smooth competition and abrupt category shifts, and that these flexibly adapt to the social context.


Sites of clonidine action to inhibit gut propulsion in mice: demonstration of a central component.
The role of central (supraspinal and spinal) and peripheral alpha-adrenoceptors in the regulation of gastrointestinal propulsion in the mouse was studied using clonidine, an alpha 2-adrenoceptor agonist. Clonidine produced a dose-dependent inhibition of propulsion when given intracerebroventricularly, intrathecally, or subcutaneously, but was most potent when given intracerebroventricularly. The antitransit effects of centrally given clonidine were antagonized by intracerebroventricular (i.c.v.) yohimbine, but higher doses were required when this antagonist was given peripherally. Whereas i.c.v. and s.c. administration of clonidine were effective in inhibiting gut transit in spinally transected mice, intrathecal (i.th.) administration of this agonist was not. A supraspinal site of clonidine action is suggested based upon (a) the higher central to peripheral potency of clonidine; (b) the greater potency of i.c.v., compared with s.c., administration of yohimbine in blocking i.c.v. clonidine; (c) the lack of effect of i.th. administration of clonidine in spinally transected mice; and (d) the reduced potency of i.c.v., but not s.c., administration of clonidine in spinally transected mice. Additionally, a peripheral site of clonidine action is suggested by (a) the lower potency of i.c.v. yohimbine in blocking s.c., compared with i.c.v., clonidine; (b) the lower potency of i.c.v. yohimbine in blocking i.c.v. clonidine in transected mice (compared with normal mice); (c) the equal potency of s.c. clonidine in slowing propulsion in normal and spinally transected mice; and (d) the equal potency of i.c.v. yohimbine in blocking s.c. clonidine in normal and spinally transected mice. These data in mice would thus support the concept that normal (peripheral) therapeutic administration of clonidine would affect gut motor function by interactions within the brain and directly at the level of the gut.


A solitary hemangioblastoma of the posterior brain fossa: the role of radiotherapy.
We report here the case of a patient admitted for management of posterior fossa cerebral hemangioblastoma. A 16-year-old male patient with a history of intracranial hypertension syndrome consisting of progressively worsening headache, vomiting, especially morning and jet vomiting, and decreased visual acuity. The patient's symptomatology worsened a few days later with the appearance of a disturbance of balance with enlargement of the sustentation polygon. The patient initially benefited from a brain computed tomography (CT) scan that objectified a solidocystic process of the posterior brain fossa. The patient then underwent a surgical excision that was considered partial and the diagnosis of hemangioblastoma was made on the surgical specimen. Since the surgical removal was partial the patient was referred to our training where he received external radiotherapy on his hemangioblastoma of the posterior brain fossa. The patient was examined one month after the end of irradiation; he presented a spectacular improvement in his neurological symptomatology with a clear regression of balance disorders. The standard treatment for cerebellar hemangioblastoma is complete microsurgical removal, but our results show a high level of efficacy for fractional photon radiotherapy after partial surgery of this benign tumour.


Acetylcholinesterase (AChE)-positive innervation of the guinea-pig spleen.
The presence and intraorgan distribution of the acetylcholinesterase (AChE)-positive nerve structures in the guinea-pig spleen were studied by means of the direct thiocholine method. Visualized AChE-positive nerve fibres entered the guinea-pig spleen at its hilum in the vicinity of the splenic artery branches and intra parenchyma were gradually distributed to form thicker periarterial nerves and also fine adventitial nerve plexus. In described topography the AChE-positive nerve fibres were identified in association with the central artery running through the white pulp. Some of the perivascular nerve fibres associated with the central artery extended away and passed into the periarterial lymphatic sheath (PALS) to reach the marginal zone and in continuation entered into the mantle zone of lymphatic follicles. Several AChE-positive nerve fibres were seen in the red pulp but less in the splenic capsule. We did not find any AChE-positive nerve cells in the guinea-pig spleen.


The transition between strong and weak chaos in delay systems: Stochastic modeling approach.
We investigate the scaling behavior of the maximal Lyapunov exponent in chaotic systems with time delay. In the large-delay limit, it is known that one can distinguish between strong and weak chaos depending on the delay scaling, analogously to strong and weak instabilities for steady states and periodic orbits. Here we show that the Lyapunov exponent of chaotic systems shows significant differences in its scaling behavior compared to constant or periodic dynamics due to fluctuations in the linearized equations of motion. We reproduce the chaotic scaling properties with a linear delay system with multiplicative noise. We further derive analytic limit cases for the stochastic model illustrating the mechanisms of the emerging scaling laws.


On-farm quality assurance programs: a survey of producer and industry leader opinions.
To assess interest in implementing a California dairy quality assurance program, practices and opinions of dairy producers and industry leaders were surveyed by a mailed questionnaire and by focus groups. The majority of the 55 participants in the focus group were dairy producers; processor marketing executives, extension dairy advisors, packinghouse executives, and dairy veterinarians were represented. The consensus among the focus groups was that a quality assurance program should be voluntary, be managed by creameries, and confer an economic advantage to participants. Focus group members listed chemical and microbial food safety (in both meat and milk), environmental health, and animal welfare as issues that should be addressed. Of the 1440 questionnaires mailed with producers' milk checks, 413 were returned. Information was collected regarding opinions and practices pertaining to administration of drugs to animals, medical records and animal identification, culling practices, manure management, cow welfare, and feeding of animal protein. An overwhelming 99% of producers believed they were responsible for the safety of meat and milk leaving the farm. Sixty percent of producers said that they would consider joining a California-specific quality assurance program, whereas 9% indicated that they would not. Producers would be more likely to join if their processor believed it would impart a market advantage and if the program standards were controlled by producers.


Modulation of CB1 cannabinoid receptor by allosteric ligands: Pharmacology and therapeutic opportunities.
Cannabinoid pharmacology has been intensely studied because of cannabis' pervasive medicinal and non-medicinal uses as well as for the therapeutic potential of cannabinoid-based drugs for the treatment of pain, anxiety, substance abuse, obesity, cancer and neurodegenerative disorders. The identification of allosteric modulators of the cannabinoid receptor 1 (CB1) has given a new direction to the development of cannabinoid-based therapeutics due to the many advantages offered by targeting allosteric site(s). Allosteric receptor modulators hold potential to develop subtype-specific and pathway-specific therapeutics. Here we briefly discuss the first-generation of allosteric modulators of CB1 receptor, their structure-activity relationships, signaling pathways and the allosteric binding site(s) on the CB1 receptor. This article is part of the Special Issue entitled "A New Dawn in Cannabinoid Neurobiology".


Infant Breastfeeding and Kidney Function in School-Aged Children.
Early life factors may influence kidney growth and function throughout the life course. We examined the associations of breastfeeding duration and exclusivity and age at introduction of solid foods with kidney outcomes at school age. Prospective cohort study from fetal life onward. 5,043 children in the Netherlands. Infant feeding was assessed prospectively using questionnaires. In children at a median age of 6.0 years, we measured kidney volume with ultrasound, estimated glomerular filtration rate (eGFR) from serum creatinine level, and microalbuminuria from urinary albumin and creatinine levels. 92% of all children were ever breastfed, of whom 27% were breastfed for more than 6 months and 21% were breastfed exclusively for at least 4 months. Compared with ever-breastfed children, never-breastfed children had smaller combined kidney volumes (-2.69 [95% CI, -4.83 to -0.56] cm(3)) and lower eGFRs (-2.42 [95% CI, -4.56 to -0.28] mL/min/1.73 m(2)) at school age. Among breastfed children, shorter duration of breastfeeding was associated with smaller combined kidney volume and lower microalbuminuria risk (P<0.05). Compared to exclusive breastfeeding for 4 months, nonexclusive breastfeeding in the first 4 months was associated with smaller combined kidney volume and lower eGFR (both P<0.05). Associations with eGFR were explained largely by kidney volume. Age at introduction of solid foods was not associated with any kidney outcome. Observational study, so causality cannot be established. Follow-up measurements were available for 76% of children. These results suggest that breastfeeding is associated with subclinical changes in kidney outcomes in childhood. Further studies are needed to explore whether early life nutrition also affects the risk of kidney disease in adulthood.


Heterotopic ossification after reverse total shoulder arthroplasty.
Heterotopic ossification (HO) occurs after arthroplasty, especially total hip arthroplasty. In this study we describe the incidence, evolution, morphology and clinical consequences of HO following reverse shoulder arthroplasty. This is a single-centre retrospective study of 132 consecutive patients who received a Delta III or Delta Xtend reverse total shoulder arthroplasty between 2006 and 2013 for the treatment of cuff tear arthropathy. There were 96 women and 36 men. Their mean age at the time of surgery was 69 years (49 to 89) and the mean follow-up was 36 months (12 to 84). The incidence, evolution, morphology and clinical consequences of HO using the Constant-Murley score (CS) were analysed. A modified Brooker classification of HO of the hip was used. HO was seen in 39 patients (29.5%). A total of 31 of these patients (81.6%) began to develop HO by three months post-operatively. According to the Hamada classification, 11 patients had grade 1a, eight had grade 1b, six had grade 1c and 14 had grade 2 HO. The HO evolved over a mean of 8.3 months (3 to 21). Patients with HO had a lower mean CS at three (p = 0.017), six (p < 0.001) and 12 months (p < 0.001) post-operatively. HO was not associated with notching (p = 0.675). HO after reverse shoulder arthroplasty is a non-progressive condition without long-term clinical consequences. Only grade 2 HO is clinically relevant with a negative effect on the function of the shoulder during its development. Cite this article: Bone Joint J 2016;98-B:1215-21.


Adenosine inhibits the excitatory synaptic inputs to Basal forebrain cholinergic, GABAergic, and parvalbumin neurons in mice.
Coffee and tea contain the stimulants caffeine and theophylline. These compounds act as antagonists of adenosine receptors. Adenosine promotes sleep and its extracellular concentration rises in association with prolonged wakefulness, particularly in the basal forebrain (BF) region involved in activating the cerebral cortex. However, the effect of adenosine on identified BF neurons, especially non-cholinergic neurons, is incompletely understood. Here we used whole-cell patch-clamp recordings in mouse brain slices prepared from two validated transgenic mouse lines with fluorescent proteins expressed in GABAergic or parvalbumin (PV) neurons to determine the effect of adenosine. Whole-cell recordings were made from BF cholinergic neurons and from BF GABAergic and PV neurons with the size (>20 μm) and intrinsic membrane properties (prominent H-currents) corresponding to cortically projecting neurons. A brief (2 min) bath application of adenosine (100 μM) decreased the frequency but not the amplitude of spontaneous excitatory postsynaptic currents (EPSCs) in all groups of BF cholinergic, GABAergic, and PV neurons we recorded. In addition, adenosine decreased the frequency of miniature EPSCs in BF cholinergic neurons. Adenosine had no effect on the frequency of spontaneous inhibitory postsynaptic currents in cholinergic neurons or GABAergic neurons with large H-currents but reduced them in a group of GABAergic neurons with smaller H-currents. All effects of adenosine were blocked by a selective, adenosine A1 receptor antagonist, cyclopentyltheophylline (CPT, 1 μM). Adenosine had no postsynaptic effects. Taken together, our work suggests that adenosine promotes sleep by an A1 receptor-mediated inhibition of glutamatergic inputs to cortically projecting cholinergic and GABA/PV neurons. Conversely, caffeine and theophylline promote attentive wakefulness by inhibiting these A1 receptors in BF thereby promoting the high-frequency oscillations in the cortex required for attention and cognition.


From single-cell variability to population growth.
Single-cell experiments have revealed cell-to-cell variability in generation times and growth rates for genetically identical cells. Theoretical models relating the fluctuating generation times of single cells to the population growth rate are usually based on the assumption that the generation times of mother and daughter cells are uncorrelated. This assumption, however, is inconsistent with the exponential growth of cell volume in time observed for many cell types. Here we develop a more general and biologically relevant model in which cells grow exponentially and generation times are correlated in a manner which controls cell size. In addition to the fluctuating generation times, we also allow the single-cell growth rates to fluctuate and account for their correlations across the lineage tree. Surprisingly, we find that the population growth rate only depends on the distribution of single-cell growth rates and their correlations.


Time trends of selected persistent organic pollutants in lake sediments from Greenland.
Sediments from seven lakes in West Greenland were used as natural archives to study past and present levels of PCBs (polychlorinated biphenyls, tri- to decachlorinated), tetra-BDE #47 (2,2',4,4'-bromodiphenyl ether), chlordane (cis- and trans-octachlordane) and HxCBz (hexachlorobenzene). The concentrations found are lower than or comparable to concentrations found in sediments from other Arctic regions and one to 2 orders of magnitude lower than concentrations typically found in sediments at lower latitudes. The observed temporal trends (direct and indirect dating) show a decreasing total PCB concentration. Even though local contamination sources exist, the POP deposition in the studied area is most likely a result from long-range transport. The hypothesis about "cold condensation" suggests a latitudinal fractionation to occur between different volatile compounds during the transport toward the pole. In this study a time delay in the deposition for the low-chlorinated PCBs (tri- and tetrachlorinated), compared to their emission histories and compared to higher chlorinated PCBs, was indicated. Although very low tetra-BDE #47 concentrations are observed in this study, there are indications for an increasing concentration in recent sediment layers that may reflect increasing environmental concentrations at lower latitudes. The investigated pesticides are still in use at lower latitudes, however neither chlordane nor HxCBz show any distinct temporal trend of increasing or decreasing concentration toward the sediment surface.


Empirical assessment of competency to consent to psychiatric hospitalization.
The authors used an interview questionnaire to assess competency to consent to voluntary admission in a group of 50 newly admitted psychiatric patients. Using a number of alternative definitions of competency with this sample of patients tested shortly after admission, they found that a majority of patients appeared to have severe impairment of competency. These results suggest limitations in the applicability of the legal model of informed consent to the psychiatric setting.


Anodal and cathodal pulsed electrical stimulation on skin wound healing in guinea pigs.
We investigated the effects of anodal and cathodal electrical stimulation on wound healing. In a randomized controlled trial, we divided 42 male albino guinea pigs into two control (C1 and C2) and four experimental (E1-E4) groups. A 3 cm linear incision was made at the dorsal skin of all guinea pigs. A unidirectional pulse current of 300 to 600 microamperes, 80 pps, and 0.3 ms pulse duration was administered for 1 hour a day. In groups E1 and E3 (anodal), a positive polarity was applied for the first 3 days followed by negative polarity the remaining days. In groups E2 and E4 (cathodal), negative polarity was applied for the first 3 days and positive polarity the remaining days. Groups E1, E2, and C1 were killed on day 14 and E3, E4, and C2 on day 21. We measured the percentage of decrease in wound surface area (daily tracing) and tensile strength (on days 14 and 21). The results indicated that both cathodal and anodal stimulations increased the rate of wound closure. Beginning with day 12, we saw a significant difference in the percentage of the decrease in wound surface between all treatment and control groups (p < 0.05). Ultimate tensile strength and stress increased in the anodal compared with the cathodal and control groups; at the end of day 14, ultimate tensile stress in E1 was significantly greater compared with C1 (p < 0.05). We conclude that electrical stimulation, regardless of polarity regimen, benefits wound healing, but anodal stimulation the first 3 days and cathodal stimulation the remaining days can lead to stronger repaired tissue.


[Brain-gut interactions].
Our digestive tract has an autonomous functioning but also has a bidirectional relation with our brain known as brain-gut interactions. This communication is mediated by the autonomous nervous system, i.e., the sympathetic and parasympathetic nervous systems, with a mixed afferent and efferent component, and the circumventricular organs located outside the blood-brain barrier. The vagus nerve, known as the principal component of the parasympathetic nervous system, is a mixed nerve composed of 90% afferent fibers, which has physiological roles due to its putative vegetative functions. The vagus nerve has also anti-inflammatory properties both through the hypothalamic pituitary adrenal axis (through its afferents) and the cholinergic anti-inflammatory pathway (through its efferents). The sympathetic nervous system has a classical antagonist effect on the parasympathetic nervous system at the origin of an equilibrated sympathovagal balance in normal conditions. The brain is able to integrate inputs coming from the digestive tract inside a central autonomic network organized around the hypothalamus, limbic system and cerebral cortex (insula, prefrontal, cingulate) and in return to modify the autonomic nervous system and the hypothalamic pituitary adrenal axis in the frame of physiological loops. A dysfunction of these brain-gut interactions, favoured by stress, is most likely involved in the pathophysiology of digestive diseases such as irritable bowel syndrome or even inflammatory bowel diseases. A better knowledge of these brain-gut interactions has therapeutic implications in the domain of pharmacology, neurophysiology, behavioural and cognitive management.


Understanding political radicalization: The two-pyramids model.
This article reviews some of the milestones of thinking about political radicalization, as scholars and security officials struggled after 9/11 to discern the precursors of terrorist violence. Recent criticism of the concept of radicalization has been recognized, leading to a 2-pyramids model that responds to the criticism by separating radicalization of opinion from radicalization of action. Security and research implications of the 2-pyramids model are briefly described, ending with a call for more attention to emotional experience in understanding both radicalization of opinion and radicalization of action. (PsycINFO Database Record


HPV E6 oncoproteins and nucleic acids in neck lymph node fine needle aspirates and oral samples from patients with oropharyngeal squamous cell carcinoma.
Commercial assays measuring HPV E6 viral oncoproteins, E6/E7 mRNA or DNA were used to test neck lymph node fine needle aspirates (FNA) and oropharyngeal samples (saliva and oral swabs) from 59 Canadian patients with oropharyngeal squamous cell carcinomas (OPSCC). Overall agreements of p16 antigen staining of tumors to FNA tested for OncoE6™, Aptima HPV E6/E7 mRNA and cobas HPV DNA were 81.4% (k 0.53), 94.9% (k 0.83) and 91.1% (k 0.73) respectively. Using HPV presence in a subset of 25 tumors as the comparator, overall agreement was 64.0% (k 0.08) with OncoE6™, 88.0% (k 0.65) with Aptima HPV E6/E7 mRNA and 91.7% (k 0.70) with cobas HPV DNA. HPV testing of oropharyngeal samples yielded lower agreements with tumor markers; 23.7-24.0% (k 0.02), 55.9-68.0% (k 0.24-0.37) and 78.9-86.9% (k 0.49-0.58) in the 3 respective tests. HPV 16 was present in 93.7-100% of the samples tested and showed 100% genotype agreement between FNA and tumors. The high rates for HPV E6 oncoproteins and E6/E7 mRNA suggests most patients were experiencing transcriptionally active HPV-related OPSCC. Results from these commercial assays performed on FNA but not oropharyngeal samples showed moderate to very good agreements with p16 and HPV testing of tumors.


Analysis of saphenous vein graft lesion composition using near-infrared spectroscopy and intravascular ultrasonography with virtual histology.
To examine the composition of saphenous vein graft (SVG) lesions using two novel modalities, near-infrared spectroscopy (NIRS) and intravascular ultrasonography with virtual histology (IVUS-VH). We performed NIRS and IVUS-VH imaging of 23 SVGs in 21 patients undergoing clinically-indicated angiography. Mean patient and SVG age was 66±7 and 10±7 years, respectively. SVG lesion location was aorto-ostial in 8 (35%), body in 13 (57%) and distal anastomotic in 2 (9%). Compared to anastomotic lesions, body lesions had larger mean lumen area (6.4±1.8 mm2 vs. 4.2±6.4 mm2, P=0.02) but similar mean plaque burden (73±5% vs. 70±10%, P=0.66). A NIRS lipid core plaque was identified in 9 of 13 body lesions vs. 1 of 10 anastomotic lesions (69% vs. 10%, P=0.005). SVG body lesions had higher lipid core burden index (LCBI) compared to anastomotic lesions (184±76 vs. 49±54, P<0.001). By IVUS-VH, SVG lesions had high % necrotic core (28±10%) and % dense calcium (13±10%), without any significant difference between body and anastomotic sites. Older SVG age was associated with higher lesion and vessel LCBI (r=0.76 and r=0.64, respectively, P<0.001), but was not associated with IVUS-VH determined plaque composition. Higher HDL-cholesterol was associated with lower lesion LCBI (r=-0.43, P=0.04). NIRS-measured lipid core plaque in SVGs increases with increasing SVG age and is infrequent in anastomotic lesions. No association was found between IVUS-VH plaque composition measurements and SVG lesion location or age.


Temperature controlled formation of polar copper phosphonates showing large dielectric anisotropy and a dehydration-induced switch from ferromagnetic to antiferromagnetic interactions.
The reaction of copper(ii) chloride with 2-carboxyphenylphosphonic acid (2-cppH3) under hydrothermal conditions at 100 °C results in the formation of a polar compound, Cu3(2-cpp)2(H2O)5 (1), which shows large dielectric anisotropy. A similar reaction at 140 °C results in a centrosymmetric phase, Cu3(2-cpp)2(H2O)2 (2). Compound 1 can convert into 2 upon increasing the reaction temperature under hydrothermal conditions, and further to Cu3(2-cpp)2 (3) upon thermal treatment at 240 °C. This process is accompanied by a switch from ferromagnetic in 1 to antiferromagnetic interactions in 2 and 3.


Broca's area network in language function: a pooling-data connectivity study.
Modern neuroimaging developments have demonstrated that cognitive functions correlate with brain networks rather than specific areas. The purpose of this paper was to analyze the connectivity of Broca's area based on language tasks. A connectivity modeling study was performed by pooling data of Broca's activation in language tasks. Fifty-seven papers that included 883 subjects in 84 experiments were analyzed. Analysis of Likelihood Estimates of pooled data was utilized to generate the map; thresholds at p < 0.01 were corrected for multiple comparisons and false discovery rate. Resulting images were co-registered into MNI standard space. A network consisting of 16 clusters of activation was obtained. Main clusters were located in the frontal operculum, left posterior temporal region, supplementary motor area, and the parietal lobe. Less common clusters were seen in the sub-cortical structures including the left thalamus, left putamen, secondary visual areas, and the right cerebellum. Broca's area-44-related networks involved in language processing were demonstrated utilizing a pooling-data connectivity study. Significance, interpretation, and limitations of the results are discussed.


Inhibitory Effects of Scutellaria baicalensis Root Extract on Linoleic Acid Hydroperoxide-induced Lung Mitochondrial Lipid Peroxidation and Antioxidant Activities.
In this study, we evaluated the ability of Scutellaria baicalensis Georgi to protect lipid-peroxidation (LPO) in lung tissue after free radical-induced injury. We prepared S. baicalensis root (SBR) extracts using different solvents. The total flavonoid and total phenol contents of each extract were measured, and the ROS damage protection was evaluated by analyzing linoleic acid hydroperoxide (LHP)-induced LPO in rat lung mitochondria. Moreover, evaluating diphenylpicrylhydrazyl (DPPH), hydrogen peroxide, superoxide anion radical, and hydroxyl radical scavenging abilities and using metal chelating assays were used to determine in vitro antioxidant activity. The ethyl acetate (EtOAc) extract showed high ROS scavenging ability, and four compounds were subsequently isolated and purified from this extract: baicalin, baicalein, wogonin, and oroxylin A. Baicalein in rat lung mitochondria the most significant LHP-induced LPO inhibition was shown and extracted with EtOAc that contained the highest amount of baicalein. Thus, baicalein and the EtOAc extract of SBR may be efficient in conferring ROS damage protection and inhibiting LHP-induced LPO in rat lung mitochondria. Additional studies are warranted to investigate their use as antioxidant therapy for respiration infections, nutrition supplements, and lead compounds in pharmaceuticals.


SKILLS - A psychoeducational group programme for children with ADHD.
Psychoeducation is regarded as a first line of intervention for attention-deficit/hyperactivity disorder (ADHD). Previous studies have been limited to psychoeducation for adult patients or parents of children with ADHD. Therefore, the aim of the present study was to investigate the acceptability and effectiveness of a psychoeducational group intervention for children with ADHD. The two-session psychoeducational intervention, SKILLS, was completed by 125 children (6-12 years) with ADHD and their parents. Self-ratings and parental ratings of ADHD symptoms, functional impairment and attitudes to diagnosis and treatment were collected before and after treatment. Post-treatment client satisfaction and parental responses to an open question about SKILLS were assessed. The data were analyzed using a combination of quantitative and qualitative (content analysis) methods. A majority of the participants expressed satisfaction with the group intervention. The intervention had no effect on symptoms and level of function, although the parents were more positive to their child's diagnosis after the intervention. The parents were positive to the group format and to the opportunity for their children to meet other children with similar symptoms. Parents wished for more interactive elements and more opportunities for children to share experiences. We concluded that the group intervention was accepted by most participants, although more activating exercises and opportunities for interaction between participants should be included. Since the intervention group was not compared with a control group, the results should be interpreted with caution. Future studies should use a randomized control treatment design and investigate the effect on adherence to later treatment.


Identification in vitro of an endothelial cell surface cofactor for antithrombin III. Parallel studies with isolated perfused rat hearts and microcarrier cultures of bovine endothelium.
Two in vitro systems were used to identify an antithrombin III cofactor activity on vascular endothelium. Langendorff rat heart preparations or columns packed with endothelium cultured on microcarrier beads were perfused with mixtures of purified thrombin and antithrombin III. With each preparation, accelerated inhibition of thrombin by antithrombin III occurred during passage over endothelium. Platelet factor 4, protamine sulfate and diisopropylphosphoryl thrombin, all antagonists of the antithrombin III cofactor activity of heparin, significantly reduced the capacity of the preparation to inhibit thrombin. It is concluded that a substance with the functional properties of a stationary phase cofactor for antithrombin III is present on the microvascular endothelium and there catalyzes the inactivation of circulating free thrombin.


Mitofusin 1 and 2 play distinct roles in mitochondrial fusion reactions via GTPase activity.
The mammalian homologues of yeast and Drosophila Fzo, mitofusin (Mfn) 1 and 2, are both essential for mitochondrial fusion and maintenance of mitochondrial morphology. Though the GTPase domain is required for Mfn protein function, the molecular mechanisms of the GTPase-dependent reaction as well as the functional division of the two Mfn proteins are unknown. To examine the function of Mfn proteins, tethering of mitochondrial membranes was measured in vitro by fluorescence microscopy using green fluorescence protein- or red fluorescent protein-tagged and Mfn1-expressing mitochondria, or by immunoprecipitation using mitochondria harboring HA- or FLAG-tagged Mfn proteins. These experiments revealed that Mfn1-harboring mitochondria were efficiently tethered in a GTP-dependent manner, whereas Mfn2-harboring mitochondria were tethered with only low efficiency. Sucrose density gradient centrifugation followed by co-immunoprecipitation revealed that Mfn1 produced oligomerized approximately 250 kDa and approximately 450 kDa complexes in a GTP-dependent manner. The approximately 450 kDa complex contained oligomerized Mfn1 from distinct apposing membranes (docking complex), whereas the approximately 250 kDa complex was composed of Mfn1 present on the same membrane or in the membrane-solubilized state (cis complex). These results were also confirmed using blue-native PAGE. Mfn1 exhibited higher activity for this reaction than Mfn2. Purified recombinant Mfn1 exhibited approximately eightfold higher GTPase activity than Mfn2. These findings indicate that the two Mfn proteins have distinct activities, and suggest that Mfn1 is mainly responsible for GTP-dependent membrane tethering.


Cause-specific mortality and comorbid neurodevelopmental disorder in 167,515 patients with bipolar disorder: An entire population longitudinal study.
Studies addressing premature mortality in bipolar disorder (BD) patients are limited by small sample sizes. Herein, we used almost 99 % of the population of Taiwan to address this issue, and its association with comorbid neurodevelopmental disorders and severe BD. Between 2003 and 2017, we enrolled 167,515 individuals with BD and controls matched 1:4 for sex and birth year from the National Health Insurance Database linked to the Database of National Death Registry in Taiwan. Time-dependent Cox regression models were used to examine cause-specific mortality (all-cause, natural, and unnatural causes [accidents or suicide]). With adjustments of sex, age, income, urbanization, and physical conditions, suicide was associated with the highest risk of mortality (reported as hazard ratio with 95 % confidence interval: 9.15; 8.53-9.81) among BD patients, followed by unnatural (4.94; 4.72-5.17), accidental (2.15; 1.99-2.32), and natural causes (1.02; 1.00-1.05). Comorbid attention-deficiency hyperactivity disorder did not contribute to the increased risk of cause-specific mortality; however, comorbid autism spectrum disorder (ASD) increased such risks, particularly for natural (3.00; 1.85-4.88) and accidental causes (7.47; 1.80-31.1). Cause-specific mortality revealed a linear trend with the frequency of psychiatric hospitalization (all, p for trend <0.001), and BD patients hospitalized twice or more each year had 34.63-fold increased risk of suicide mortality (26.03-46.07). BD patients with a higher frequency of psychiatric hospitalization have the highest risk of suicide mortality, and comorbid ASD was associated with an increased risk of natural and accidental causes of mortality.


Effect of Vetiveria zizanioides essential oil on melanogenesis in melanoma cells: downregulation of tyrosinase expression and suppression of oxidative stress.
The major objective of this study was to estimate the hypopigmentation function of the essential oil from Vetiveria zizanioides (VZ-EO). Our results indicated that VZ-EO exhibits potent lipid peroxidation inhibitory activity to moderate the bleaching of β-carotene and to maintain the cellular glutathione (GSH) levels. VZ-EO can markedly decrease melanin production and tyrosinase activity in α-melanin-stimulating-hormone- (α-MSH-) stimulated B16 cells. The effect of VZ-EO on melanogenesis is achieved by the suppression of cellular tyrosinase expression. The results demonstrated that the activity of VZ-EO on melanogenesis might be the result of its potent antioxidative ability, which was reflected in the decreased cellular oxidant and malondialdehyde (MDA) levels and the recovered activities of superoxide dismutase (SOD), glutathione peroxidase (GPX), and catalase (CAT) in α-MSH-stimulated B16 cells. The most abundant compound in VZ-EO is cedr-8-en-13-ol (12.4%), which has a strong capability to inhibit lipid peroxidation. Therefore, VZ-EO has the potential to become an ingredient in future hypopigmentation drugs, foods, and cosmetics.


Formation pathway of Roussin's red ester (RRE) via the reaction of a {Fe(NO)2}(10) dinitrosyliron complex (DNIC) and thiol: facile synthetic route for synthesizing cysteine-containing DNIC.
Transformation of {Fe(NO)2}(10) dinitrosyliron complex (DNIC) Fe(CO)2(NO)2 into [{Fe(NO)2}(9)]2 Roussin's red ester (RRE) [(μ-S(CH2)2NH2)Fe(NO)2]2 (3) triggered by cysteamine via the reaction pathway (intermediates) [{Fe(NO)2}(10)]2[(NO)2Fe(μ-CO)(μ-S(CH2)2NH3)Fe(NO)2] (1) → {Fe(NO)2}(9){Fe(NO)2}(10)[(NO)2Fe(μ-S(CH2)2NH2)(μ-S(CH2)2NH3)Fe(NO)2] (2) → RRE 3 is demonstrated. The 1-to-2-to-3 conversion is promoted by proton transfer followed by O2 oxidation and deprotonation. Additionally, a study on facile conversion of complex 3 to complexes [(SR)(S(CH2)2NH3)Fe(NO)2] [SR = 2-aminoethanethiolate (4), benzenethiolate (5)] and [(CysS))(S(CH2)2NH3)Fe(NO)2] (6) via reaction with thiols and the further utility of complex 5 as a template for synthesizing mixed-thiolate-containing reduced RRE (rRRE) [(μ-SC6H5)(μ-S(CH2)2NH3)Fe2(NO)4] (7) provide the methodology for the synthesis and isolation of neutral, pure cysteine-/mixed-thiolate-containing DNIC/RRE. Compared to the conversion of complex 2 to complex 3 via reaction with O2, diphenyl disulfide triggers oxidation of complex 2 to lead to formation of the neutral {Fe(NO)2}(9) DNIC 5 and RRE 3. S-S bond activation of diphenyl disulfide by rRRE 2 may support the decay (oxidation) of rRRE species in ToMOC via the reduction of adjacent protein residues such as cystins, proposed by Lippard.


A large-scale survey on finger counting routines, their temporal stability and flexibility in educated adults.
A strong link between bodily activity and number processing has been established in recent years. Although numerous observations indicate that adults use finger counting (FC) in various contexts of everyday life for different purposes, existing knowledge of FC routines and their use is still limited. In particular, it remains unknown how stable the (default) FC habits are over time and how flexible they can be. To investigate these questions, 380 Polish participants completed a questionnaire on their FC routines, the stability of these routines, and the context of FC usage, preceded by the request to count on their fingers from 1 to 10. Next, the test-retest stability of FC habits was examined in 84 participants 2 months following the first session. To the best of our knowledge, such a study design has been adopted for the first time. The results indicate that default FC routines of the majority of participants (75%) are relatively stable over time. At the same time, FC routines can flexibly adapt according to the situation (e.g., when holding an object). As regards prevalence, almost all participants, in line with previous findings on Western individuals, declared starting from the closed palm and extending consecutive fingers. Furthermore, we observed relations between FC preferences and handedness (more left-handers start from the left hand) and that actual finger use is still widespread in healthy adults for a variety of activities (the most prevalent uses of FC are listing elements, presenting arguments and plans, and calendar calculations). In sum, the results show the practical relevance of FC in adulthood, the relative stability of preferences over time along with flexible adaptation to a current situation, as well as an association of FC routines with handedness. Taken together our results suggest that FC is the phenomenon, which is moderated or mediated by multiple embodied factors.


Potassium citrate prevents increased osteoclastogenesis resulting from acidic conditions: Implication for the treatment of postmenopausal bone loss.
The extracellular acidic milieu in bones results in activation of osteoclasts (OC) and inhibition of osteoblasts (OB) causing a net loss of calcium from the skeleton and the deterioration of bone microarchitecture. Alkalinization through supplementation with potassium citrate (K citrate) has been proposed to limit the osteopenia progression, even though its pharmacological activity in bone microenvironment is not well defined. We evaluated if K citrate was able to prevent the adverse effects that acidic milieu induces on bone cells. OC and OB were maintained in neutral (pH 7.4) versus acidic (pH 6.9) culture medium, and treated with different K citrate concentrations. We evaluated the OC differentiation at seven days, by counting of multinucleated cells expressing tartrate-resistant acid phosphatase, and the activity of mature OC at 14 days, by quantifying of collagen degradation. To evaluate the effects on OB, we analyzed proliferation, mineralization, and expression of bone-related genes. We found that the low pH increased OC differentiation and activity and decreased OB function. The osteoclastogenesis was also promoted by RANKL concentrations ineffective at pH 7.4. Non-cytotoxic K citrate concentrations were not sufficient to steadily neutralize the acidic medium, but a) inhibited the osteoclastogenesis, the collagen degradation, and the expression of genes involved in RANKL-mediated OC differentiation, b) enhanced OB proliferation and alkaline phosphatase expression, whereas it did not affect the in vitro mineralization, and c) were effective also in OC cultures resistant to alendronate, i.e. the positive control of osteoclastogenesis inhibition. In conclusion, K citrate prevents the increase in OC activity induced by the acidic microenvironment, and the effect does not depend exclusively on its alkalizing capacity. These data provide the biological basis for the use of K citrate in preventing the osteopenia progression resulting from low-grade acidosis.


Prognostic value of the expression of p53, bcl-2, and bax oncoproteins, and neovascularization in patients with radically resected non-small-cell lung cancer.
To assess the prognostic value of p53, bcl-2, bax, and neovascularization in radically resected non-small-cell lung cancer (NSCLC) patients. Tumors from 116 patients were assessed by immunohistochemistry for expression of p53 (DO7 and PAb1081), bcl-2, and the quantification of microvessel density (CD-31). In addition, the expression of bax was assessed in 61 stage I tumors. The median levels of expression of each marker were used as cutoff points. p53 was not correlated to any patient or tumor characteristic, whereas bcl-2 showed higher expression in squamous cell carcinomas (P < .001). bax expression was significantly related with male sex (P = .006) and adenocarcinoma type (P = .0013). p53 status, assessed with one monoclonal antibody (MoAb), was not predictive for survival; however, the combination of staining results obtained with two MoAbs identified the DO7-/PAb1801+ tumors as those with the worst prognosis. bcl-2 expression was associated with longer survival in stage I patients (P = .0169). The combined group expressing p53+(PAb1801)/bcl-2- had the worst survival in stage I patients (P = .034) and in the whole series in comparison with the other combinations of the two oncoproteins. bax expression alone had no influence on survival of stage I patients, but patients with bax+/bcl-2- tumors had the worst prognosis (P = .02 in comparison with bax+/bcl-2+). Tumor neovascularization was not related with other factors, and patients with CD-31+ tumors had a shorter survival duration than those with CD-31- tumors only in stage II (P = .0283). By multivariate analysis including all patients, the presence of p53+/ bcl-2- tumor expression and large tumor diameter (> or = 4cm) were independent prognostic factors for shorter survival duration. For stage I, only the presence of bax+/ bcl-2- tumor expression had a significant negative influence on survival. The interaction and the regulation of new biologic markers, such as those involved in the apoptotic pathway, are complex. Combinations of the expression of several of them may give more valuable information than the study of just one. Prognostic influence of p53 staining varied depending on the choice of antibody and the combination of bcl-2- together with p53+ (PAb1801) or with bax+ had the worst influence on survival for patients with stage I NSCLC.


[Refeeding syndrome. A review].
Refeeding syndrome is a complex clinical picture that encompass all those alterations that can occur as a consequence of the nutritional support (oral, enteral or parenteral) in malnourished patients. Refeeding syndrome is classically characterized by neurological alterations, respiratory symptoms, cardiac arrhythmias and heart failure few days after beginning of refeeding, with life-threatening outcome. Its pathogenesis includes alterations in the corporal fluids, and in some electrolytes, minerals and vitamins. In this article a review of refeeding syndrome pathogenesis and clinical manifestations is carried out, with a final series of recommendations for lowering the risk of this syndrome and for facilitate the early diagnosis and the treatment.


Systematic instruction of arthroscopic knot tying with the ArK Trainer: an objective evaluation tool.
A Proficiency Formula was introduced as an objective self-evaluation method for evaluating basic arthroscopic knot tying in a laboratory setting. The correlation between the Proficiency Formula and gold standard pass/fail dichotomy was demonstrated, as well as with other popular evaluation tools--task-specific checklist (TSC) and global rating scale (GRS). A step-by-step video tutorial was used to instruct 35 medical students on how to tie an arthroscopic Samsung Medical Center (SMC) knot secured by three half hitches. Participants were video recorded performing arthroscopic knot tying and assessed on their success tying an SMC knot, pass or fail, and through three outcome tools: the Proficiency Formula, GRS and the TSC. Independent samples t test was used to compare the GRS, TSC and Proficiency Formula scores, between those who were passed or failed by the evaluators. Correlation between the measurement scales was tested using Spearman's rho correlation coefficient. Participants received a mean proficiency score of 195 (140-249). The mean Proficiency score for those that passed was 323 (95 % CI 272-374), for those that failed, 87 (95 % CI 26-148, p < 0.001). We found strong linear correlation between the Proficiency Formula and GRS and TSE (0.83 and 0.78, respectively). The Proficiency Formula has high correlation with gold standard GRS and TSC measurements when used to assess arthroscopic knot tying skills on a model. It has the added advantage of being able to be self-assessed.


Community-acquired pneumonia: still a major burden of disease.
Describe recent studies that may impact on the management of community-acquired pneumonia (CAP). CAP continues to be associated with a considerable burden of disease. Diagnosis remains problematic, and various biomarkers are neither accurate in the diagnosis of the presence of CAP nor superior to standard severity of illness scores in predicting outcome. Current evidence indicates that patients with nonsevere CAP can be effectively treated with antibiotic monotherapy, whereas those with severe infection, particularly ICU cases, do best with early initiation of combination antibiotic therapy. Several studies have investigated anti-inflammatory, adjunctive therapies for severe CAP, with corticosteroids appearing to be most promising. It is well recognized that cardiac complications occur during the course of CAP, being associated with poorer short-term and long-term outcomes, prompting considerable interest in the adjunctive potential of statins and antiplatelet therapies. In addition to evaluating these adjunctive therapies, attention has also focused on identifying strategies that predict the need for ICU admission in patients with CAP. Although questions remain, particularly with regard to prediction of outcome, recent studies of CAP, both clinical and experimental, have contributed novel insights into disease pathogenesis that may enable improvement of current treatment strategies.


Risk factors for invasive fungal infections in patients with connective tissue disease: Systematic review and meta-analysis.
Invasive fungal infections (IFIs) are life-threatening opportunistic infections in patients with connective tissue disease CTD) that cause significant morbidity and mortality. We attempted to determine the potential risk factors associated with IFIs in CTD. We systematically searched PubMed, Embase, and the Cochrane Library databases for relevant articles published from the database inception to February 1, 2023. Twenty-six studies were included in this systematic review and meta-analysis. Risk factors identified for IFIs were diabetes (odds ratio [OR], 1.62; 95% confidence interval [CI], 1.00 to 2.64), pulmonary diseases (OR 3.43; 95% CI 2.49 to 4.73), interstitial lung disease (ILD; OR, 4.06; 95% CI, 2.22 to 7.41), renal disease (OR, 4.41; 95% CI, 1.84 to 10.59), glucocorticoid (GC) use (OR, 4.15; 95% CI, 2.74 to 6.28), especially moderate to high-dose GC, azathioprine (AZA) use (OR, 1.50; 95% CI, 1.12 to 2.01), calcineurin inhibitor (CNI) use (OR, 2.49; 95% CI, 1.59 to 3.91), mycophenolate mofetil (MMF) use (OR, 2.83; 95% CI, 1.59 to 5.03), cyclophosphamide (CYC) use (OR, 3.35; 95% CI, 2.47 to 4.54), biologics use (OR, 3.43; 95% CI, 2.36 to 4.98), and lymphopenia (OR, 4.26; 95% CI, 2.08 to 8.73). Hydroxychloroquine (HCQ) use reduced risk of IFIs (OR, 0.67; 95% CI, 0.54 to 0.84). Furthermore, 17 of the 26 studies only reported risk factors for Pneumocystis jiroveci pneumonia (PJP) in patients with CTD. Pulmonary disease; ILD; and the use of GC, CNIs, CYC, methotrexate (MTX), MMF and biologics, and lymphopenia increased the risk of PJP, whereas the use of HCQ reduced its risk. Diabetes, pulmonary disease, ILD, renal disease, use of GC (especially at moderate to high dose) and immunosuppressive drugs, and lymphopenia were found to be associated with significant risk for IFIs (especially PJP) in patients with CTD. Furthermore, the use of HCQ may reduce the risk of IFIs in patients with CTD.


Development of assays to support identification and characterization of modulators of DExH-box helicase DHX9.
DHX9 is a DExH-box RNA helicase that utilizes hydrolysis of all four nucleotide triphosphates (NTPs) to power cycles of 3' to 5' directional movement to resolve and/or unwind double stranded RNA, DNA, and RNA/DNA hybrids, R-loops, triplex-DNA and G-quadraplexes. DHX9 activity is important for both viral amplification and maintaining genomic stability in cancer cells; therefore, it is a therapeutic target of interest for drug discovery efforts. Biochemical assays measuring ATP hydrolysis and oligonucleotide unwinding for DHX9 have been developed and characterized, and these assays can support high-throughput compound screening efforts under balanced conditions. Assay development efforts revealed DHX9 can use double stranded RNA with 18-mer poly(U) 3' overhangs and as well as significantly shorter overhangs at the 5' or 3' end as substrates. The enzymatic assays are augmented by a robust SPR assay for compound validation. A mechanism-derived inhibitor, GTPγS, was characterized as part of the validation of these assays and a crystal structure of GDP bound to cat DHX9 has been solved. In addition to enabling drug discovery efforts for DHX9, these assays may be extrapolated to other RNA helicases providing a valuable toolkit for this important target class.


Persistent high prevalence of cardiovascular risk factors in the urban middle class in India: Jaipur Heart Watch-5.
Urban subjects have high burden of cardiovascular risk factors, therefore, to evaluate risk factors in middle socioeconomic subjects and to study secular trends we performed an epidemiological study. The study was performed at urban middle class locations defined according to municipal records in years 2009-10. Stratified random sampling using house-to-house survey was performed. Details of medical history, anthropometry and clinical examination were recorded and biochemical tests performed for estimation of fasting glucose and lipids. Current definitions were used for risk factor classification. Descriptive statistics are provided. Trends were calculated using ANOVA or Mantel Haenszel chi-square. Univariate and multivariate logistic regression was performed to assess risk factor determinants. To determine secular trends we compared risk factors with previous cross-sectional studies performed in same locations in years 2002-3 and 2004-5 in subjects 20-59 years age. We evaluated 739 subjects (men 451, women 288, response 67%). Age-adjusted prevalence (%) of risk factors in men and women respectively was smoking 95 (21.1) and 12 (4.2), low physical activity 316 (69.6) and 147 (52.3), high fat intake > or = 20 gm/day 278 (73.4) and 171 (68.7), low fruits and vegetables intake < 3 helpings/day 249 (70.3) and 165 (76.4), overweight/obesity 205 (46.2) and 142 (50.7), high waist size 58 (12.9) and 76 (26.6), high waist:hip 143 (31.9) and 154 (53.9), hypertension 177 (39.5) and 71 (24.6), high total cholesterol > or = 200 mg/ dl 148 (33.0) and 93 (32.7), low HDL cholesterol < 40/50 mg/dl 113 (25.1) and 157 (55.3), diabetes 62 (15.5) and 25 (10.8) and metabolic syndrome 109 (25.1) and 61 (22.0). Age-associated increase was observed in body mass index, waist size, waist ratio:hip, systolic blood pressure and fasting and total cholesterol, non-HDL cholesterol and triglycerides in women (Ptrend < 0.01). Age related increase was also observed in prevalence of obesity, truncal obesity, hypertension, diabetes and metabolic syndrome (Ptrend < 0.01). On univariate analysis significant determinants of risk factors were low educational and socioeconomic status for smoking, high fat diet for obesity and hypertension, low fruits and vegetables intake for metabolic syndrome, and low physical activity or obesity but on age-and sex-adjusted multivariate analysis only association was high fat diet with obesity and hypertension (logistic regression analysis p < 0.05). Compared to studies performed at similar locations in years 2002-03 and 2005-06 there was increasing trend in prevalence of high non-HDL cholesterol and hypertriglyceridemia (Ptrend < 0.05) while other risk factors did not change significantly. There is a high prevalence of multiple cardiovascular risk factors in Indian middle class individuals. Secular trends demonstrate a persistent high prevalence and increasing non-HDL cholesterol and triglycerides over 8-year period.


Simple methods of determining confidence intervals for functions of estimates in published results.
Often, the reader of a published paper is interested in a comparison of parameters that has not been presented. It is not possible to make inferences beyond point estimation since the standard error for the contrast of the estimated parameters depends upon the (unreported) correlation. This study explores approaches to obtain valid confidence intervals when the correlation [Formula: see text] is unknown. We illustrate three proposed approaches using data from the National Health Interview Survey. The three approaches include the Bonferroni method and the standard confidence interval assuming [Formula: see text] (most conservative) or [Formula: see text] (when the correlation is known to be non-negative). The Bonferroni approach is found to be the most conservative. For the difference in two estimated parameter, the standard confidence interval assuming [Formula: see text] yields a 95% confidence interval that is approximately 12.5% narrower than the Bonferroni confidence interval; when the correlation is known to be positive, the standard 95% confidence interval assuming [Formula: see text] is approximately 38% narrower than the Bonferroni. In summary, this article demonstrates simple methods to determine confidence intervals for unreported comparisons. We suggest use of the standard confidence interval assuming [Formula: see text] if no information is available or [Formula: see text] if the correlation is known to be non-negative.


Prognostic value of components of body composition in patients treated with targeted therapy for advanced renal cell carcinoma: a retrospective case series.
To evaluate the association between various components of body composition and overall survival of patients treated with targeted therapies for advanced renal cell carcinoma. This retrospective study included 124 Chinese patients with advanced renal cell carcinoma who had been treated with targeted therapy from 2008 to 2012 at Fudan University Cancer Center. The L3 plane from a computed tomography scan was analyzed. Area and density were recorded as quantitative and quality measures. Univariate and multivariate Cox proportion hazard regression models were constructed to calculate the crude and adjusted hazard ratio (HR) of various components of body composition for overall survival. X-tile software was used to search for optimal cutoffs for male and female patients and the concordance index evaluated incremental changes in prognostication. After adjusting for age, sex and Heng risk stratification, only visceral adipose tissue index (HR 0.981, p = 0.002) and subcutaneous adipose tissue index (HR 0.987, p = 0.048) were independently associated with overall survival. Visceral adipose tissue remained a significant prognostic factor (HR 0.997, p = 0.005) when the influence of body mass index was included. Using defined cutoffs, patients with low VAT had double the death rate (p = 0.007). Visceral adipose tissue also added significant benefit to Heng risk stratification. Further exploratory analysis revealed that visceral adipose tissue may be an indicator of nutritional status in patients with advanced renal cell carcinoma. Radiologic measurement of VAT is an independent prognostic factor for Asian patients treated with targeted therapy for advanced renal cell carcinoma.


Biological and clinical roles of IL-18 in inflammatory diseases.
Several new discoveries have revived interest in the pathogenic potential and possible clinical roles of IL-18. IL-18 is an IL-1 family cytokine with potent ability to induce IFNγ production. However, basic investigations and now clinical observations suggest a more complex picture. Unique aspects of IL-18 biology at the levels of transcription, activation, secretion, neutralization, receptor distribution and signalling help to explain its pleiotropic roles in mucosal and systemic inflammation. Blood biomarker studies reveal a cytokine for which profound elevation, associated with detectable 'free IL-18', defines a group of autoinflammatory diseases in which IL-18 dysregulation can be a primary driving feature, the so-called 'IL-18opathies'. This impressive specificity might accelerate diagnoses and identify patients amenable to therapeutic IL-18 blockade. Pathogenically, human and animal studies identify a preferential activation of CD8+ T cells over other IL-18-responsive lymphocytes. IL-18 agonist treatments that leverage the site of production or subversion of endogenous IL-18 inhibition show promise in augmenting immune responses to cancer. Thus, the unique aspects of IL-18 biology are finally beginning to have clinical impact in precision diagnostics, disease monitoring and targeted treatment of inflammatory and malignant diseases.


A transient inward current elicited by hyperpolarization during serotonin activation in Xenopus oocytes.
Activation of serotonin, glutamate or muscarinic receptors, incorporated into the membrane of Xenopus oocytes following injection of messenger RNA from rat brain, caused the development of a transient inward (Tin) current when the membrane was hyperpolarized. A detailed study was made of the Tin current induced during serotonin activation. The current is due principally to efflux of chloride ions, and is presumably activated by an influx of calcium ions, because it was blocked by removal of calcium from the bathing medium, by addition of manganese, cobalt or lanthanum, or by intracellular injection of EGTA. During application of serotonin, the amplitude of the Tin current increased slowly, and after washing it persisted for longer than the direct serotonin-induced current. The amplitude of the Tin current was sensitive to temperature and pH, and was abolished at pH 6.5 or by cooling to 12 degrees C. The Tin current may be of importance in regulating the excitability of neurons in the central nervous system.


[Lessons from the organization of medical support during the Battle for Moscow].
Covered and analyzed the state of medical support during the Battle for Moscow. The problems of organizing of medical support for troops during all stages of this war--defensive operations and counter-offensive strategic operations are considered. Analyzed errors and mistakes made in the course of medical support of combat operations of the fronts that participated in the battle for Moscow. The article traces the solutions of the problems encountered and assess the results of translating these decisions into practice. Particular attention is paid to problems of organization of work as a hospital base fronts and armies, and the medical units and divisional managers, as well as management of medical service, evacuate the wounded and sick, the dynamics of formation of the provision of specialized medical care and anti security forces.


Structural basis for the inhibition of histone deacetylase 8 (HDAC8), a key epigenetic player in the blood fluke Schistosoma mansoni.
The treatment of schistosomiasis, a disease caused by blood flukes parasites of the Schistosoma genus, depends on the intensive use of a single drug, praziquantel, which increases the likelihood of the development of drug-resistant parasite strains and renders the search for new drugs a strategic priority. Currently, inhibitors of human epigenetic enzymes are actively investigated as novel anti-cancer drugs and have the potential to be used as new anti-parasitic agents. Here, we report that Schistosoma mansoni histone deacetylase 8 (smHDAC8), the most expressed class I HDAC isotype in this organism, is a functional acetyl-L-lysine deacetylase that plays an important role in parasite infectivity. The crystal structure of smHDAC8 shows that this enzyme adopts a canonical α/β HDAC fold, with specific solvent exposed loops corresponding to insertions in the schistosome HDAC8 sequence. Importantly, structures of smHDAC8 in complex with generic HDAC inhibitors revealed specific structural changes in the smHDAC8 active site that cannot be accommodated by human HDACs. Using a structure-based approach, we identified several small-molecule inhibitors that build on these specificities. These molecules exhibit an inhibitory effect on smHDAC8 but show reduced affinity for human HDACs. Crucially, we show that a newly identified smHDAC8 inhibitor has the capacity to induce apoptosis and mortality in schistosomes. Taken together, our biological and structural findings define the framework for the rational design of small-molecule inhibitors specifically interfering with schistosome epigenetic mechanisms, and further support an anti-parasitic epigenome targeting strategy to treat neglected diseases caused by eukaryotic pathogens.


Regeneration in the ctenophore Mnemiopsis leidyi occurs in the absence of a blastema, requires cell division, and is temporally separable from wound healing.
The ability to regenerate is a widely distributed but highly variable trait among metazoans. A variety of modes of regeneration has been described for different organisms; however, many questions regarding the origin and evolution of these strategies remain unanswered. Most species of ctenophore (or "comb jellies"), a clade of marine animals that branch off at the base of the animal tree of life, possess an outstanding capacity to regenerate. However, the cellular and molecular mechanisms underlying this ability are unknown. We have used the ctenophore Mnemiopsis leidyi as a system to study wound healing and adult regeneration and provide some first-time insights of the cellular mechanisms involved in the regeneration of one of the most ancient extant group of multicellular animals. We show that cell proliferation is activated at the wound site and is indispensable for whole-body regeneration. Wound healing occurs normally in the absence of cell proliferation forming a scar-less wound epithelium. No blastema-like structure is generated at the cut site, and pulse-chase experiments and surgical intervention show that cells originating in the main regions of cell proliferation (the tentacle bulbs) do not seem to contribute to the formation of new structures after surgical challenge, suggesting a local source of cells during regeneration. While exposure to cell-proliferation blocking treatment inhibits regeneration, the ability to regenerate is recovered when the treatment ends (days after the original cut), suggesting that ctenophore regenerative capabilities are constantly ready to be triggered and they are somehow separable of the wound healing process. Ctenophore regeneration takes place through a process of cell proliferation-dependent non-blastemal-like regeneration and is temporally separable of the wound healing process. We propose that undifferentiated cells assume the correct location of missing structures and differentiate in place. The remarkable ability to replace missing tissue, the many favorable experimental features (e.g., optical clarity, high fecundity, rapid regenerative performance, stereotyped cell lineage, sequenced genome), and the early branching phylogenetic position in the animal tree, all point to the emergence of ctenophores as a new model system to study the evolution of animal regeneration.


Exonuclease III (XthA) Enforces In Vivo DNA Cloning of Escherichia coli To Create Cohesive Ends.
Escherichia coli has an ability to assemble DNA fragments with homologous overlapping sequences of 15 to 40 bp at each end. Several modified protocols have already been reported to improve this simple and useful DNA cloning technology. However, the molecular mechanism by which E. coli accomplishes such cloning is still unknown. In this study, we provide evidence that the in vivo cloning of E. coli is independent of both RecA and RecET recombinases but is dependent on XthA, a 3' to 5' exonuclease. Here, in vivo cloning of E. coli by XthA is referred to as in vivoE. coli cloning (iVEC). We also show that iVEC activity is reduced by deletion of the C-terminal domain of DNA polymerase I (PolA). Collectively, these results suggest the following mechanism of iVEC. First, XthA resects the 3' ends of linear DNA fragments that are introduced into E. coli cells, resulting in exposure of the single-stranded 5' overhangs. Then, the complementary single-stranded DNA ends hybridize each other, and gaps are filled by DNA polymerase I. Elucidation of the iVEC mechanism at the molecular level would further advance the development of in vivo DNA cloning technology. Already we have successfully demonstrated multiple-fragment assembly of up to seven fragments in combination with an effortless transformation procedure using a modified host strain for iVEC.IMPORTANCE Cloning of a DNA fragment into a vector is one of the fundamental techniques in recombinant DNA technology. Recently, an in vitro recombination system for DNA cloning was shown to enable the joining of multiple DNA fragments at once. Interestingly, E. coli potentially assembles multiple linear DNA fragments that are introduced into the cell. Improved protocols for this in vivo cloning have realized a high level of usability, comparable to that by in vitro recombination reactions. However, the mechanism of in vivo cloning is highly controversial. Here, we clarified the fundamental mechanism underlying in vivo cloning and also constructed a strain that was optimized for in vivo cloning. Additionally, we streamlined the procedure of in vivo cloning by using a single microcentrifuge tube.


SPECT assessment of brain activation induced by caffeine: no effect on areas involved in dependence.
Caffeine is not considered addictive, and in animals it does not trigger metabolic increases or dopamine release in brain areas involved in reinforcement and reward. Our objective was to measure caffeine effects on cerebral perfusion in humans using single photon emission computed tomography with a specific focus on areas of reinforcement and reward. Two groups of nonsmoking subjects were studied, one with a low (8 subjects) and one with a high (6 subjects) daily coffee consumption. The subjects ingested 3 mg/kg caffeine or placebo in a raspberry-tasting drink, and scans were performed 45 min after ingestion. A control group of 12 healthy volunteers receiving no drink was also studied. Caffeine consumption led to a generalized, statistically nonsignificant perfusion decrease of 6% to 8%, comparable in low and high consumers. Compared with controls, low consumers displayed neuronal activation bilaterally in inferior frontal gyrus-anterior insular cortex and uncus, left internal parietal cortex, right lingual gyrus, and cerebellum. In high consumers, brain activation occurred bilaterally only in hypothalamus. Thus, on a background of widespread low-amplitude perfusion decrease, caffeine activates a few regions mainly involved in the control of vigilance, anxiety, and cardiovascular regulation, but does not affect areas involved in reinforcing and reward.


In vivo virulence of MHC-adapted AIDS virus serially-passaged through MHC-mismatched hosts.
CD8+ T-cell responses exert strong suppressive pressure on HIV replication and select for viral escape mutations. Some of these major histocompatibility complex class I (MHC-I)-associated mutations result in reduction of in vitro viral replicative capacity. While these mutations can revert after viral transmission to MHC-I-disparate hosts, recent studies have suggested that these MHC-I-associated mutations accumulate in populations and make viruses less pathogenic in vitro. Here, we directly show an increase in the in vivo virulence of an MHC-I-adapted virus serially-passaged through MHC-I-mismatched hosts in a macaque AIDS model despite a reduction in in vitro viral fitness. The first passage simian immunodeficiency virus (1pSIV) obtained 1 year after SIVmac239 infection in a macaque possessing a protective MHC-I haplotype 90-120-Ia was transmitted into 90-120-Ia- macaques, whose plasma 1 year post-infection was transmitted into other 90-120-Ia- macaques to obtain the third passage SIV (3pSIV). Most of the 90-120-Ia-associated mutations selected in 1pSIV did not revert even in 3pSIV. 3pSIV showed lower in vitro viral fitness but induced persistent viremia in 90-120-Ia- macaques. Remarkably, 3pSIV infection in 90-120-Ia+ macaques resulted in significantly higher viral loads and reduced survival compared to wild-type SIVmac239. These results indicate that MHC-I-adapted SIVs serially-transmitted through MHC-I-mismatched hosts can have higher virulence in MHC-I-matched hosts despite their lower in vitro viral fitness. This study suggests that multiply-passaged HIVs could result in loss of HIV-specific CD8+ T cell responses in human populations and the in vivo pathogenic potential of these escaped viruses may be enhanced.


Effects of protein supplementation on lean body mass, muscle strength, and physical performance in nonfrail community-dwelling older adults: a systematic review and meta-analysis.
Increasing protein intake has been suggested as an effective strategy to ameliorate age-related loss of muscle mass and strength. Current reviews assessing the effect of protein supplementation are strongly influenced by the inclusion of studies with frail older adults. We assessed the effect of protein supplementation on lean body mass, muscle strength, and physical performance in exclusively nonfrail community-dwelling older adults. Moreover, we assessed the superior effects of protein supplementation during concomitant resistance exercise training on muscle characteristics. A systematic literature search was conducted on PubMed, Embase, and Web of Science up to 15 May 2018. We included randomized controlled trials that assessed the effect of protein supplementation on lean body mass, muscle thigh cross-sectional area, muscle strength, gait speed, and chair-rise ability and performed random-effects meta-analyses. Data from 36 studies with 1682 participants showed no significant effects of protein supplementation on changes in lean body mass [standardized mean difference (SMD): 0.11; 95% CI: -0.06, 0.28], handgrip strength (SMD: 0.58; 95% CI: -0.08, 1.24), lower extremity muscle strength (SMD: 0.03; 95% CI: -0.20, 0.27), gait speed (SMD: 0.41; 95% CI: -0.04, 0.85), or chair-rise ability (SMD: 0.10; 95%: CI -0.08, 0.28) compared with a control condition in nonfrail community-dwelling older adults. Moreover, no superior effects of protein supplementation were found during concomitant resistance exercise training on muscle characteristics. Protein supplementation in nonfrail community-dwelling older adults does not lead to increases in lean body mass, muscle cross-sectional area, muscle strength, or physical performance compared with control conditions; nor does it exert superior effects when added to resistance exercise training. Habitual protein intakes of most study participants were already sufficient, and protein interventions differed in terms of type of protein, amount, and timing. Future research should clarify what specific protein supplementation protocol is beneficial for nonfrail community-dwelling older adults with low habitual protein intake.


Syntheses of Gold and Silver Nanocomposite Contact Lenses via Chemical Volumetric Modulation of Hydrogels.
Integration of nanomaterials into hydrogels has emerged as a prominent research tool utilized in applications such as sensing, cancer therapy, and bone tissue engineering. Wearable contact lenses functionalized with nanoparticles have been exploited in therapeutics and targeted therapy. Here, we report the fabrication of gold and silver nanocomposite commercial contact lenses using a breathing-in/breathing-out (BI-BO) method, whereby a hydrated contact lens is shrunk in an aprotic solvent and then allowed to swell in an aqueous solution containing nanoparticles. The morphology and optical properties of the gold and silver nanoparticles were characterized through transmission electron microscopy and ultraviolet-visible spectroscopy. The transmission spectra of nanocomposite contact lenses indicated that the nanoparticles' loading amount within the lens depended primarily on the number of BI-BO cycles. Nanocomposites were stable for a minimum period of 1 month, and no nanoparticle leaching was observed. Wettability and water content analysis of the nanocomposites revealed that the contact lenses retained their intrinsic material properties after the fabrication process. The dispersion of the nanoparticles within the contact lens media was determined through scanning electron microscopy imaging. The nanocomposite lenses can be deployed in color filtering and antibacterial applications. In fact, the silver nanocomposite contact lens showed blue-light blocking capabilities by filtering a harmful high-energy blue-light range (400-450 nm) while transmitting the visible light beyond 470 nm, which facilitates enhanced night vision and color distinction. The ease of fabricating these nanocomposite contact lenses via the BI-BO method could enable the incorporation of nanoparticles with diverse morphologies into contact lenses for various biomedical applications.


Spasticity Management: The Current State of Transcranial Neuromodulation.
This narrative review aims to provide an objective view of the noninvasive neuromodulation (NINM) protocols available for treating spasticity, including repetitive transcranial magnetic stimulation (rTMS) and transcranial direct current stimulation (tDCS). On the basis of the relevant randomized controlled trials, we infer that NINM is more effective in reducing spasticity when combined with the conventional therapies than used as a stand-alone treatment. However, the magnitude of NINM after-effects depends significantly on the applied hemisphere and the underlying pathology. Being in line with these arguments, low-frequency rTMS and cathodal-tDCS over the unaffected hemisphere are more effective in reducing spasticity than high-frequency rTMS and anodal-tDCS over the affected hemisphere in chronic poststroke. However, most of the studies are heterogeneous in the stimulation setup, patient selection, follow-up duration, and the availability of the sham operation. Therefore, the available data on the usefulness of NINM in reducing spasticity need to be confirmed by larger and multicentric randomized controlled trials to gather evidence on the efficiency of NINM regimens in reducing spasticity in various neurologic conditions. V.


Characterization of gag and pol sequences from long-term survivors of human immunodeficiency virus type 1 infection.
We previously identified a group of 10 long-term survivors (LTS) of human immunodeficiency virus type 1 (HIV-1) infection. Extensive biological analysis revealed that some of these individuals do well, at least in part, because they possess weakened or attenuated viruses. Also, previously, to determine the genotype associated with the attenuated phenotype in vivo, we characterized nef, vif, vpr, vpu, env, and LTR in our cohort of LTS. In this study, we analyzed gag and pol genes derived from eight individuals in our cohort. For each subject multiple full-length gag and pol clones were obtained for analysis. In most cases, the sequences derived from the LTS had an intact open reading frame. At the protein level, there were no discernible differences between the sequences derived from LTS and those derived from patients with AIDS. Thus, no common defect in gag and pol was found in our cohort. One individual (subject SF), however, had only Gag-defective proviral sequences (10 of 10) in his peripheral blood mononuclear cells. Furthermore, longitudinal studies of the samples collected from SF over a 2-year period showed that all p17 gag clones sequenced (24 of 24) were defective due to G-to-A hypermutations. This viral defect in Gag may provide the molecular basis for this individual's extremely low viral load and long-term asymptomatic state. These results, together with previous findings in our LTS cohort, reinforce the notion that it is unlikely that a single common viral genetic determinant accounts for the lack of disease progression in all cases. Multiple host and viral factors undoubtedly contribute to the well-being of LTS of HIV-1 infection.


Why do some patients with type 1 diabetes live so long?
While the lifespan of people with type 1 diabetes has increased progressively since the advent of insulin therapy, these patients still experience premature mortality, primarily from cardiovascular disease (CVD). However, a subgroup of those with type 1 diabetes survives well into old age without significant morbidity. It is the purpose of this review to explore the factors which may help in identifying these patients. It might be expected that hyperglycaemia plays a major role in explaining the increased incidence of CVD and mortality of these individuals. However, while a number of publications have associated poor long term glycaemic control with an increase in both all-cause mortality and CVD in those with type 1 diabetes, it is apparent that good glycaemic control alone cannot explain why some patients with type 1 diabetes avoid fatal CVD events. Lipid disorders may occur in those with type 1 diabetes, but the occurrence of elevated high-density lipoprotein-cholesterol is positively associated with longevity in this population. Non-renal hypertension, by itself is a significant risk factor for CVD but if adequately treated does not appear to mitigate against longevity. However, the presence of nephropathy is a major risk factor and its absence after 15-20 years of diabetes appears to be a marker of long-term survival. One of the major factors linked with long-term survival is the absence of features of the metabolic syndrome and more specifically the presence of insulin sensitivity. Genetic factors also play a role, with a family history of longevity and an absence of type 2 diabetes and hypertension in the family being important considerations. There is thus a complex interaction between multiple risk factors in determining which patients with type 1 diabetes are likely to live into older age. However, these patients can often be identified clinically based on a combination of factors as outlined above.


Frequency of self-monitoring of blood glucose in patients with type 2 diabetes: association with hypoglycaemic events.
The objective of this study is to describe current self-monitoring of blood glucose (SMBG) practice for patients with type 2 diabetes by treatment type and adherence with healthcare professional advice concerning SMBG. In addition, the study aims to investigate the association of SMBG and self-reported episodes of low blood glucose. This cross-sectional survey design study was carried out on patients with type 2 diabetes aged 18 years or over, attending community pharmacies in 97 sites across the United Kingdom. Patients picking up a prescription for blood glucose test strips or diabetes medicine from a community pharmacist were asked to complete a questionnaire. The pharmacist was available to assist if requested. Questions included: self-reports of frequency of blood glucose testing; type of diabetes treatment; advice given by healthcare professionals about frequency of blood glucose testing; frequency of episodes of low blood glucose; and last known HbA(1c) level. The final sample size was 554 respondents, who were grouped for analysis as follows: those being treated with insulin, either alone or with any oral medication (n = 167); those being treated with sulfonylureas, either alone or with any oral medication (n = 187); and those being treated with any other medication, or controlled by diet and exercise alone (n = 202). Frequency of SMBG was higher in patients using insulin (median 10 times per week, Q (1), Q (3) = 4.5, 14) than in patients on treatments other than insulin (four times per week, Q (1), Q (3) = 2, 7, p < 0.001). SMBG was carried out at the same frequency in patients not treated with insulin regardless of whether they were prescribed sulfonylureas. Greater frequency of SMBG was associated with self-reports of one or more episodes of low blood glucose in the previous six months. Among patients with type 2 diabetes, those treated with insulin used SMBG at a greater frequency than those not treated with insulin. Increased frequency of testing was associated with increased frequency of self-reported episodes of low blood glucose, even among patients not taking insulin or sulfonylureas. This raises the possibility that episodes of hypoglycaemia may not be accurately identified, leading to unnecessary fear, or conversely that treatment is not being adjusted to avoid such morbidity. Although further work is needed to explore this association in a representative, prospective cohort of patients, possible explanations for reports of low-blood glucose should be discussed with patients using SMBG more frequently to ensure they are able to accurately identify episodes of hypoglycaemia.


Nonpigmented dysplastic melanocytic nevi.
Dysplastic melanocytic nevi (DMN) are thought to represent a clinical and histologic bridge between common pigmented nevi and superficial spreading malignant melanoma. The following clinical criteria for DMN were established to aid in the proper identification of these lesions: irregular perimeter, size exceeding 5 mm in diameter, background erythema, and variegated color (shades of browns, tans, blacks, and reds). Histologic features include basilar melanocytic proliferation with nuclear atypia, a patchy lymphocytic infiltrate with concentric eosinophilic fibroplasia, and lamellar fibroplasia. To our knowledge, there have been no previously reported cases of uniformly nonpigmented DMN. A 31-year-old brown-haired, browneyed white woman with no personal or family history of either DMN or melanoma presented for evaluation of numerous, discrete, nonindurated, 2- to 5-mm-diameter, nonpigmented macules and slightly elevated papules that had appeared in a truncal distribution over the course of several years. Microscopic examination of these lesions showed lentiginous epidermal hyperplasia and disordered proliferation with variable cellular atypia of intraepidermal melanocytes. Nonpigmented, nonindurated, macular or slightly elevated papular lesions may represent nevi with features of dysplasia. In light of the significant risk of malignant melanoma that is associated with pigmented varieties of dysplastic nevi, it is essential that clinicians consider nonpigmented DMN in the differential diagnosis of entities that present as hypopigmented macules.


Changes over time in digital literacy.
The current study is a follow-up on the 2002 empirical study by Eshet-Alkalai and Amichai-Hamburger, which investigated digital literacy skills among different age groups. This study explores changes through time in digital literacy among the same participants 5 years later, and their performance is compared to new matched control groups. Results indicate an improvement over time among all age groups, but especially for the adults, in the tasks that require proficiency and technical control in using technology (e.g., photovisual and branching literacy skills). On the other hand, results indicate a drop in the skills that require creative and critical thinking (e.g., information and reproduction literacy skills), especially for the younger participants. Results show two major patterns of change over time: (a) closing the gap between younger and older participants in the tasks that emphasize proficiency and technical control and (b) widening the gap between younger and older participants in tasks that emphasize creativity and critical thinking. Based on the comparison with the matched control groups, we suggest that experience with technology, and not age, accounts for the observed lifelong changes in digital literacy skills.


The European Association for Cardio-Thoracic Surgery (EACTS) database: an introduction.
Continuous monitoring of surgical outcomes through benchmarking and the identification of best practices has become increasingly important. A structured approach to data collection, coupled with validation, analysis and reporting, is a powerful tool in these endeavours. However, inconsistencies in standards and practices have made comparisons within and between European countries cumbersome. The European Association for Cardio-Thoracic Surgery (EACTS) has established a large international database with the goals of (i) working with other organizations towards universal data collection and creating a European-wide repository of information on the practice of cardio-thoracic surgery, and (ii) disseminating that information in scientific, peer-reviewed articles. We report on the process of data collection, as well as on an overview of the data in the database. The EACTS Database Committee met for the first time in Monaco, September 2002, to establish the ground rules for the process of setting up the database. Subsequently, data have been collected and merged by Dendrite Clinical Systems Ltd. As of December 2008, the database included 1,074,168 patient records from 366 hospitals located in 29 countries. The latest submission from the years 2006-08 included 404,721 records. The largest contributors were the UK (32.0%), Germany (20.9%) and Belgium (7.3%). Isolated coronary bypass surgery was the most frequently performed operation; the proportion of surgical workload that comprised isolated coronary artery bypass grafting varied from country to country: 30% in Spain and almost 70% in Denmark. Isolated valve procedures constituted 12% of all procedures in Norway and 32% in Spain. Baseline demographics showed an increase in the mean age and the percentage of patients that were female over time. Remarkably, the mortality rates for all procedures declined over the period analysed, to 2.2% (95% confidence interval [CI] 2.2-2.3%) for isolated coronary bypass, 3.4% (95% CI 3.3-3.5%) for isolated valve and 6.2% (95% CI 6.0-6.5%) for bypass + valve procedures. The EACTS database has proven to be an important step forward in providing opportunities for monitoring cardiac surgical care across Europe. As the database continues to expand, it will facilitate research projects, establish benchmarking standards and identify potential areas for quality improvements.


Conformational dynamics of DNA hairpins at millisecond resolution obtained from analysis of single-molecule FRET histograms.
Here we provide high resolution study of DNA hairpin dynamics achieved by probability distribution analysis (PDA) of diffusion-based single-molecule Förster resonance energy transfer (sm-FRET) histograms. The opening and closing rates of three hairpins both free and attached to DNA origami were determined. The agreement with rates previously obtained using the total internal reflection (TIRF) technique and between free hairpins and hairpins attached to origami validated the PDA and demonstrated that the origami had no influence on the hairpin dynamics. From comparison of rates of four DNA hairpins, differing only in stem sequence, and from comparison with rates calculated using nearest-neighbor method and standard transition state theory, we conclude that the unfolding reaction resembles that of melting of DNA duplex with a corresponding sequence and that the folding reaction depends on counterion concentration and not on stem sequence. Our validation and demonstration of the PDA method will encourage its implementation in future high-resolution dynamic studies of freely diffusing biomolecules.