Comparing variation across European countries: building geographical areas to provide sounder estimates.
In geographical studies, population distribution is a key issue. An unequal distribution across units of analysis might entail extra-variation and produce misleading conclusions on healthcare performance variations. This article aims at assessing the impact of building more homogeneous units of analysis in the estimation of systematic variation in three countries. Hospital discharges for six conditions (congestive heart failure, short-term complications of diabetes, hip fracture, knee replacement, prostatectomy in prostate cancer and percutaneous coronary intervention) produced in Denmark, England and Portugal in 2008 and 2009 were allocated to both original geographical units and new ad hoc areas. New areas were built using Ward's minimum variance methods. The impact of the new areas on variability was assessed using Kernel distribution curves and different statistic of variation such as Extremal Quotient, Interquartile Interval ratio, Systematic Component of Variation and Empirical Bayes statistic. Ward's method reduced the number of areas, allowing a more homogeneous population distribution, yet 20% of the areas in Portugal exhibited less than 100 000 inhabitants vs. 7% in Denmark and 5% in England. Point estimates for Extremal Quotient and Interquartile Interval Ratio were lower in the three countries, particularly in less prevalent conditions. In turn, the Systematic Component of Variation and Empirical Bayes statistic were slightly lower in more prevalent conditions. Building new geographical areas produced a reduction of the variation in hospitalization rates in several prevalent conditions mitigating random noise, particularly in the smallest areas and allowing a sounder interpretation of the variation across countries.


Detection of mRNAs of carcinoembryonic antigen and nonspecific cross-reacting antigen genes in colorectal adenomas and carcinomas by in situ hybridization.
Expression of mRNAs of carcinoembryonic antigen (CEA) and nonspecific cross-reacting antigen (NCA) genes in colorectal carcinomas and adenomas was investigated by in situ hybridization (ISH) with specific biotinylated probes. Hybridization was clearly detected throughout the cytoplasm of 7 out of 15 adenomas and 13 out of 15 carcinomas with the CEA cDNA probe, and in 6 out of 15 adenomas and 10 out of 15 carcinomas with the NCA cDNA probe. The intensity of signal appeared to be stronger in carcinomas than that in adenomas, and the CEA and NCA mRNAs were expressed together in most of the positive tissue specimens. On the other hand, noninvaded tissues adjacent to the carcinoma did not show any signal except for 4 cases faintly stained with the NCA probe. This finding was partly confirmed by Northern blot analysis which indicated that the specific bands for the CEA and NCA mRNAs were more intense in RNAs from carcinoma tissues than in those from adjacent noninvaded tissues. These data suggest that the ISH technique with biotinylated probes could be of use for analyzing expression and localization of CEA and related genes on tissue sections.


Endovascular Management of a Combined Subclavian and Vertebral Artery Injury in an Unstable Polytrauma Patient: Case Report and Literature Review.
While blunt trauma of the head and neck are a common pattern of injury, significant problems related to the prompt diagnosis and optimal management of traumatic artery injuries have been reported in the literature. While patients with major artery injuries might develop hemorrhagic shock very rapidly, patients with blunt cerebrovascular injuries (BCVI) can present asymptomatic, but complications like basilar territory infarction, cortical blindness and death may occur. We report the life- and limb-saving management in a 57-year-old hemodynamically unstable trauma patient. The individual developed hemorrhagic shock, and other major complications, including cortical blindness, related to a posterior circulation stroke. Full recovery was achieved by immediate endovascular prosthesis for subclavian artery (SA) rupture and stenting of a traumatic vertebral artery occlusion. Endovascular and alternative treatment options are discussed and the management of subsequent sequelae associated with aggressive anticoagulation in trauma patients is reviewed, including intracranial, abdominal and other sites of secondary hemorrhage.


Herpes simplex virus type 1 ICP27-dependent activation of NF-kappaB.
The ability of herpes simplex virus type 1 (HSV-1) to activate NF-kappaB has been well documented. Beginning at 3 to 5 h postinfection, HSV-1 induces a robust and persistent nuclear translocation of an NF-kappaB-dependent (p50/p65 heterodimer) DNA binding activity, as measured by electrophoretic mobility shift assay. Activation requires virus binding and entry, as well as de novo infected-cell protein synthesis, and is accompanied by loss of both IkappaBalpha and IkappaBbeta. In this study, we identified loss of IkappaBalpha as a marker of NF-kappaB activation, and infection with mutants with individual immediate-early (IE) regulatory proteins deleted indicated that ICP27 was necessary for IkappaBalpha loss. Analysis of both N-terminal and C-terminal mutants of ICP27 identified the region from amino acids 21 to 63 as being necessary for IkappaBalpha loss. Additional experiments with mutant viruses with combinations of IE genes deleted revealed that the ICP27-dependent mechanism of NF-kappaB activation may be augmented by functional ICP4. We also analyzed two additional markers for NF-kappaB activation, phosphorylation of the p65 subunit on Ser276 and Ser536. Phosphorylation of both serines was induced upon HSV infection and required functional ICP4 and ICP27. Pharmacological inhibitor studies revealed that both IkappaBalpha and Ser276 phosphorylation were dependent on Jun N-terminal protein kinase activity, while Ser536 phosphorylation was not affected during inhibitor treatment. These results demonstrate that there are several layers of regulation of NF-kappaB activation during HSV infection, highlighting the important role that NF-kappaB may play in infection.


Cost-effectiveness of implementing a suicide prediction tool (OxMIS) in severe mental illness: Economic modeling study.
Cost-effectiveness analysis needs to be considered when introducing new tools and treatments to clinical services. The number of new assessment tools in mental health has rapidly expanded, including suicide risk assessment. Such suicide-based assessments, when linked to preventative interventions, are integral to high-quality mental health care for people with severe mental illness (SMI). We examined the cost implications of implementing Oxford Mental Illness and Suicide (OxMIS), an evidence-based, scalable suicide risk assessment tool that provides probabilistic estimates of suicide risk over 12 months for people with SMI in England. We developed a decision analytic model using secondary data to estimate the potential cost-effectiveness of incorporating OxMIS into clinical decision-making in secondary care as compared to usual care. Cost-effectiveness was measured in terms of costs per quality-adjusted life years (QALYs) gained. Uncertainty was addressed with deterministic and probabilistic sensitivity analysis. Conducting suicide risk assessment with OxMIS was potentially cheaper than clinical risk assessment alone by £250 (95% confidence interval, -786;31) to £599 (-1,321;-156) (in 2020-2021 prices) per person with SMI and associated with a small increase in quality of life (0.01 [-0.03;0.05] to 0.01 QALY, [-0.04;0.07]). The estimated incremental cost-effectiveness ratio of implementing OxMIS was cost saving. Using probabilistic sensitivity analysis, 99.96% of 10,000 simulations remained cost saving. Cost-effectiveness analysis can be conducted on risk prediction models. Implementing one such model that focuses on suicide risk in a high-risk population can lead to cost savings and improved health outcomes, especially if explicitly linked to preventative treatments.


Prognostic value of the human antigen R (HuR) in human breast cancer: high level predicts a favourable prognosis.
The role of human antigen R (HuR) and its prognostic value in breast cancer is currently only partially understood. This study aimed to determine the levels of expression of HuR in breast cancer and assess its prognostic value. HuR expression levels were assessed in a cohort of human breast cancer specimens and cell lines using both quantitative and qualitative analysis along with immunohistochemical techniques and the results were compared to the patient details. Immunohistochemical staining revealed a weak HuR staining pattern in breast tumour sections compared to the intense epithelial staining of the normal breast tissues. The levels of HuR transcripts were lower in the more advanced TNM4 and TNM3, poor outcome NPI-2 (Nottingham Prognostic Index) and NPI-3 and node-positive tumours compared with the early stage TNM2 and TNM1, NPI-1, and node-negative tumours, although these values did not reach statistical significance. The patients with metastasis, those who died of breast cancer and those with bone metastasis had significantly decreased levels of HuR transcripts, (p=0.031, p=0.018 or p=0.038 respectively) compared to the disease-free patients. The patients with poor prognosis, including those with metastasis and those who died of breast cancer, had a significantly lower level of HuR transcripts compared to the disease-free patients (p=0.021). High levels of HuR correlated with longer overall survival, although the values did not reach statistical significance (p=0.06). While no significant association of the levels of HuR expression with pathological status of breast cancer is reported, HuR may represent a prognostic factor in human breast cancer, as patients expressing high levels of HuR have a favourable prognosis, and the prognostic role of HuR for breast cancer metastasis, particularly bone metastasis is supported.


The SNAP trial: a randomised placebo-controlled trial of nicotine replacement therapy in pregnancy--clinical effectiveness and safety until 2 years after delivery, with economic evaluation.
Smoking during pregnancy causes many adverse pregnancy and birth outcomes. Nicotine replacement therapy (NRT) is effective for cessation outside pregnancy but efficacy and safety in pregnancy are unknown. We hypothesised that NRT would increase smoking cessation in pregnancy without adversely affecting infants. To compare (1) at delivery, the clinical effectiveness and cost-effectiveness for achieving biochemically validated smoking cessation of NRT patches with placebo patches in pregnancy and (2) in infants at 2 years of age, the effects of maternal NRT patch use with placebo patch use in pregnancy on behaviour, development and disability. Randomised, placebo-controlled, parallel-group trial and economic evaluation with follow-up at 4 weeks after randomisation, delivery and until infants were 2 years old. Randomisation was stratified by centre and a computer-generated sequence was used to allocate participants using a 1 : 1 ratio. Participants, site pharmacies and all study staff were blind to treatment allocation. Seven antenatal hospitals in the Midlands and north-west England. Women between 12 and 24 weeks' gestation who smoked ≥ 10 cigarettes a day before and ≥ 5 during pregnancy, with an exhaled carbon monoxide (CO) reading of ≥ 8 parts per million (p.p.m.). NRT patches (15 mg per 16 hours) or matched placebo as an 8-week course issued in two equal batches. A second batch was dispensed at 4 weeks to those abstinent from smoking. self-reported, prolonged abstinence from smoking between a quit date and childbirth, validated at delivery by CO measurement and/or salivary cotinine (COT) (primary outcome). Infants, at 2 years: absence of impairment, defined as no disability or problems with behaviour and development. Economic: cost per 'quitter'. One thousand and fifty women enrolled (521 NRT, 529 placebo). There were 1010 live singleton births and 12 participants had live twins, while there were 14 fetal deaths and no birth data for 14 participants. Numbers of adverse pregnancy and birth outcomes were similar in trial groups, except for a greater number of caesarean deliveries in the NRT group. Smoking: all participants were included in the intention-to-treat (ITT) analyses; those lost to follow-up (7% for primary outcome) were assumed to be smoking. At 1 month after randomisation, the validated cessation rate was higher in the NRT group {21.3% vs. 11.7%, odds ratio [OR], [95% confidence interval (CI)] for cessation with NRT, 2.05 [1.46 to 2.88]}. At delivery, there was no difference between groups' smoking cessation rates: 9.4% in the NRT and 7.6% in the placebo group [OR (95% CI), 1.26 (0.82 to 1.96)]. Infants: at 2 years, analyses were based on data from 888 out of 1010 (87.9%) singleton infants (including four postnatal infant deaths) [445/503 (88.5%) NRT, 443/507 (87.4%) placebo] and used multiple imputation. In the NRT group, 72.6% (323/445) had no impairment compared with 65.5% (290/443) in placebo (OR 1.40, 95% CI 1.05 to 1.86). The incremental cost-effectiveness ratio for NRT use was £4156 per quitter (£4926 including twins), but there was substantial uncertainty around these estimates. Nicotine replacement therapy patches had no enduring, significant effect on smoking in pregnancy; however, 2-year-olds born to women who used NRT were more likely to have survived without any developmental impairment. Further studies should investigate the clinical effectiveness and safety of higher doses of NRT. Current Controlled Trials ISRCTN07249128. This project was funded by the NIHR Health Technology Assessment programme and will be published in full in Health Technology Assessment; Vol. 18, No. 54. See the NIHR Journals Library programme website for further project information.


The information needs of women newly diagnosed with breast cancer.
Nurses can play a key role in patient education, including providing patients with useful and appropriate information. Rather than focusing on the process of education or information giving by nurses, this study places emphasis on the content of that information by taking the patients' perspective and asking the patients themselves what particular types of information are perceived as important at a specific point in time. The aim of the study was to explore what particular types of information were important to women newly diagnosed with breast cancer; to enable nurses and other health care professionals to utilize their time as effectively as possible and provide a high-quality service to individuals in their care. Women with breast cancer (a mean of 2.5 weeks from diagnosis) were interviewed and asked to compare items of information. The items of information were presented in pairs and the women stated a preference for one item in that pair. Thirty-six pairs were presented in total. The analysis involved the use of a Thurstone scaling model, which allowed rank orderings, or profiles of information needs, to be developed, reflecting the perceived importance of each item. Information about the likelihood of cure, the spread of the disease and treatment options were perceived as the most important items of information at the time of diagnosis. Other information needs, in order of descending priority, included information about the risk to family, side-effects of treatments, impact on family, self-care, effect on social life and sexual attractiveness. Profiles of information needs were produced to take account of differences in age, level of education and social class.(ABSTRACT TRUNCATED AT 250 WORDS)


The Cost of Hyperkalemia in the United States.
There are limited data on the cost of hyperkalemia. This retrospective analysis of the Truven MarketScan claims database assessed the economic burden of hyperkalemia among selected adult patients with hyperkalemia and matched controls. A total of 39,626 cases (patients with hyperkalemia) were matched to 39,626 controls (patients without hyperkalemia) based on age, dialysis, chronic kidney disease (CKD) stage, heart failure, and renin-angiotensin aldosterone system inhibitor use. Compared with controls, cases incurred $4128 (95% confidence interval [CI] $3893-$4363) higher 30-day total health care costs ($5994 vs. $1865) and $15,983 (95% CI $15,026-$16,940) higher 1-year costs ($31,844 vs. $15,861). Among 11,221 matched pairs of patients with CKD and/or heart failure, cases incurred $5553 (95% CI $5059-$6047) higher 30-day total health care costs ($8165 vs. $2612) and $24,133 (95% CI $21,748-$26,518) higher 1-year costs ($48,994 vs. $24,861) than controls. The multivariable adjusted 1-year total health care cost difference was $15,606 (95% CI $14,648-$16,576) among all patients and $25,156 (95% CI $23,529-$26,757) among patients with CKD and/or heart failure. Cases had higher resource utilization rates including inpatient admissions (30-day: 0.14 vs. 0.03; 1-year: 0.44 vs. 0.19), outpatient visits (30-day: 3.33 vs. 2.28; 1-year: 26.58 vs. 18.53), and emergency department visits (30-day: 0.16 vs. 0.06; 1-year: 0.86 vs. 0.50) (all P < 0.001). When hospitalized, cases stayed 1.51 days (95% CI 1.22-1.80) longer and were 40% more likely to be readmitted. These data indicate that hyperkalemia is associated with a significant economic burden on afflicted patients and the health care system.


Testing a flexible-receptor docking algorithm in a model binding site.
Sampling receptor flexibility is challenging for database docking. We consider a method that treats multiple flexible regions of the binding site independently, recombining them to generate different discrete conformations. This algorithm scales linearly rather than exponentially with the receptor's degrees of freedom. The method was first evaluated for its ability to identify known ligands of a hydrophobic cavity mutant of T4 lysozyme (L99A). Some 200000 molecules of the Available Chemical Directory (ACD) were docked against an ensemble of cavity conformations. Surprisingly, the enrichment of known ligands from among a much larger number of decoys in the ACD was worse than simply docking to the apo conformation alone. Large decoys, accommodated in the larger cavity conformations sampled in the ensemble, were ranked better than known small ligands. The calculation was redone with an energy correction term that considered the cost of forming the larger cavity conformations. Enrichment improved, as did the balance between high-ranking large and small ligands. In a second retrospective test, the ACD was docked against a conformational ensemble of thymidylate synthase. Compared to docking against individual enzyme conformations, the flexible receptor docking approach improved enrichment of known ligands. Including a receptor conformational energy weighting term improved enrichment further. To test the method prospectively, the ACD database was docked against another cavity mutant of lysozyme (L99A/M102Q). A total of 18 new compounds predicted to bind this polar cavity and to change its conformation were tested experimentally; 14 were found to bind. The bound structures for seven ligands were determined by X-ray crystallography. The predicted geometries of these ligands all corresponded to the observed geometries to within 0.7A RMSD or better. Significant conformational changes of the cavity were observed in all seven complexes. In five structures, part of the observed accommodations were correctly predicted; in two structures, the receptor conformational changes were unanticipated and thus never sampled. These results suggest that although sampling receptor flexibility can lead to novel ligands that would have been missed when docking a rigid structure, it is also important to consider receptor conformational energy.


Motivation to pursue genetic testing in individuals with a personal or family history of cardiac events or sudden cardiac death.
Genetic testing is becoming increasingly available for cardiac channelopathies, such as long QT syndrome and Brugada syndrome, which can lead to sudden cardiac death. Test results can be used to shape an individual's medical management and to identify at-risk family members. In our qualitative study, all participants had a personal or family history of a diagnosed cardiac arrhythmia syndrome or sudden cardiac death. Open-ended interviews were conducted individually and in focus groups. Interviews were audio recorded, transcribed verbatim, and analyzed using a qualitative grounded-theory approach. Of 50 participants, 37 described their motivations for pursuing genetic testing for long QT syndrome or another cardiac channelopathy. Participants' motivations included: to find an explanation for a family member's sudden death, to relieve uncertainty regarding a diagnosis, to guide future medical management, to allay concern about children or other family members, and to comply with recommendations of physicians or family members. Perceived reasons not to pursue genetic testing included denial, fear, and lack of information. The genetic counseling and informed consent process can be enhanced by understanding and addressing an individual's internal and external motivations either for or against pursuing genetic testing.


TpPtMe(H)(2): why is there H/D scrambling of the methyl group but not methane loss?
The reactivity of TpPtMe(H)(2) (Tp = hydrido-tris(pyrazolyl)borate) was investigated. This complex is remarkably resistant to methane loss; heating it in methanol at 55 degrees C does not lead to either methane or hydrogen loss. When CD(3)OD is used, reversible H/D scrambling of the hydrides and the methyl hydrogens occurs. This reactivity was investigated by density functional theory (DFT) methods at the mPW1k/LANL2DZ+P//mPW1k/LANL2DZ level. It was found that methane loss cannot occur due to the rigidity of the Tp ligand, which does not allow the trans geometry which would be required for the product of methane elimination, TpPtH. The resulting complex is very high in energy, and therefore the loss of methane is unfavorable. On the other hand, H/D scrambling of the methyl ligand is relatively facile. It proceeds through an eta(2-CH)-CH(4) complex, even though methane loss is not observed. The model system, [(NH(3))(3)PtMe(H)(2)](+) was examined to verify that the cause of the observations is the rigidity of the Tp system. The reaction was investigated at a number of levels of DFT. It was concluded that investigations of similar sized systems should be examined at the above level of theory or the mPW1k/SDB-cc-pVDZ//mPW1k/SDD level for improved accuracy of the energetic calculations.


MTHFR C677T homozygous mutation in a patient with pigmentary glaucoma and central retinal vein occlusion.
We present the rare case of a young male patient with asymmetric ocular findings: pigmentary ocular hypertension associated with nonischemic central retinal vein occlusion (CRVO) in the right eye and pigmentary glaucoma (PG) with progressive glaucomatous optic damage in the left eye. A 31-year-old man showed nonischemic CRVO in the right eye and the clinical triad of pigment dispersion syndrome in both eyes, however more marked in the left eye. Best-corrected visual acuity was logMAR 0.3 in the right eye and 1.0 in the left eye at presentation. The single risk for developing PG and CRVO was hyperhomocysteinemia. The patient was a carrier of the methylenetetrahydrofolate reductase C677 homozygous mutation. At 18 months of follow-up, visual acuity remained stable, intraocular pressure was in the normal range, but retinal tomography indicated an increase in glaucomatous optic damage to the nerve fiber layer in almost the complete temporal-inferior sector of the left eye, but without visual field defects in the left eye. Retinal tomography and automated perimetry were normal in the right eye. The patient received topical antiglaucomatous therapy. Higher levels of plasma homocysteine, even mildly elevated ones, could be associated with nonischemic CRVO and PG, especially when related to genetic risk factors or C677T mutation.


Depletion of foxp3(+) T cells abrogates tolerance of skin and heart allografts in murine mixed chimeras without the loss of mixed chimerism.
The relative contribution of central and peripheral mechanisms to the generation and maintenance of allograft tolerance is of considerable interest. Here, we present new evidence that regulatory T cells (Foxp3(+) ) maintain skin and heart allograft tolerance in mixed hematopoietic chimeric mice. Transient depletion of both donor- and recipient-derived Foxp3(+) cells was necessary and sufficient to induce decisive rejection of long-accepted skin and heart allografts. In contrast, stable hematopoietic chimerism remained, and there was no detectable induction of donor-specific reactivity to hematopoietic cells. Foxp3(+) cell depletion did not result in the rejection of skin grafts of only MHC-disparate donors (B6.C-H2(d) /bByJ), indicating that MHC antigens were not the target in the graft. We conclude that two different mechanisms of tolerance are present in mixed chimeras. Hematopoietic chimerism, resistant to Foxp3(+) depletion, is probably due to deletional tolerance to MHC antigens, as supported by previous studies. In contrast, regulatory tolerance mechanisms involving Foxp3(+) cells are required to control reactivity against non-MHC antigens not present on hematopoietic lineages.


[Influence of personal, professional and cross-national factors in burnout syndrome in Hispanic Americans and Spanish health workers (2007)].
Burnout syndrome is related to cultural and individual factors. The aim of this study was to compare the frequency of burnout and the scores for its three components with the perceptions and the demographic and professional characteristics of the workers. Burnout syndrome was studied in 11,530 Hispanic Americans and Spanish healthcare professionals (51% male, mean age 41.7 years). The Maslach Burnout Inventory and a previously drawn up questionnaire were administered online from the Intramed website from December 2006 to September 2007. Associations were tested using multiple logistic regression. The frequency of burnout in professionals resident in Spain was 14.9%, in Argentina 14.4%, and in Uruguay 7.9% whereas professionals in Mexico, Ecuador, Peru, Columbia, Uruguay, Guatemala and El Salvador presented frequencies of burnout of between 2.5% and 5.9%. By professions, doctors had a prevalence of burnout of 12.1%, nurses 7.2%, and dentists, psychologists and nutritionists of <6%. Amongst doctors, burnout predominated amongst doctors working in emergency departments (17%) and internal medicine departments (15.5%) whereas anaesthetists and dermatologists had the lowest prevalence (5% and 5.3%, respectively). Older age (OR=0.96), having children (OR=0.93), the perception of feeling valued (OR=0.53), optimism (OR=0.80), job satisfaction (OR=0.80), and satisfaction with salary (OR=0.91) are variables which protect against burnout. The expression of burnout varies among nations and professions. Age (older age), having children, the perception of feeling valued, optimism, job satisfaction and satisfaction with salary are protective variables of burnout.


Dopamine Modulates Option Generation for Behavior.
Animals make innumerable decisions every day, each of which involves evaluating potential options for action. But how are options generated? Although much is now known about decision making when a fixed set of potential options is provided, surprisingly little progress has been made on self-generated options. Some researchers have proposed that such abilities might be modulated by dopamine. Here, we used a new measure of option generation that is quantitative, objective, and culture fair to investigate how humans generate different behavioral options. Participants were asked to draw as many different paths (options) as they could between two points within a fixed time. Healthy individuals (n = 96) exhibited a trade-off between uniqueness (how individually different their options were) and fluency (number of options), generating either many similar or few unique options. To assess influence of dopamine, we first examined patients with Parkinson's disease (n = 35) ON and OFF their dopaminergic medication and compared them to elderly healthy controls (n = 34). Then we conducted a double-blind, placebo-controlled crossover study of the D2 agonist cabergoline in healthy older people (n = 29). Across both studies, dopamine increased fluency but diminished overall uniqueness of options generated, due to the effect of fluency trading off with uniqueness. Crucially, however, when this trade-off was corrected for, dopamine was found to increase uniqueness for any given fluency. Three carefully designed control studies showed that performance on our option-generation task was not related to executing movements, planning actions, or selecting between generated options. These findings show that dopamine plays an important role in modulating option generation.


Insulin activation of phosphatidylinositol 3-kinase in the hypothalamic arcuate nucleus: a key mediator of insulin-induced anorexia.
In peripheral tissues, insulin signaling involves activation of the insulin receptor substrate (IRS)-phosphatidylinositol 3-kinase (PI3K) enzyme system. In the hypothalamus, insulin functions with leptin as an afferent adiposity signal important for the regulation of body fat stores and hepatic glucose metabolism. To test the hypothesis that hypothalamic insulin action involves intracellular PI3K signaling, we used histochemical and biochemical methods to determine the effect of insulin on hypothalamic IRS-PI3K activity. Here, we report that insulin induces tyrosine phosphorylation of the insulin receptor and IRS-1 and -2, increases binding of activated IRS-1 and -2 to the regulatory subunit of PI3K, and activates protein kinase B/Akt, a downstream target of PI3K. Using an immunohistochemical technique to detect PI 3,4,5-triphosphate, the main product of PI3K activity, we further demonstrate that in the arcuate nucleus, insulin-induced PI3K activity occurs preferentially within cells that contain IRS-2. Finally, we show that the food intake- lowering effects of insulin are reversed by intracerebroventricular infusion of either of two PI3K inhibitors at doses that have no independent feeding effects. These findings support the hypothesis that the IRS-PI3K pathway is a mediator of insulin action in the arcuate nucleus and, combined with recent evidence that leptin activates PI3K signaling in the hypothalamus, provide a plausible mechanism for neuronal cross-talk between insulin and leptin signaling.


Calreticulin is a fine tuning molecule in epibrassinolide-induced apoptosis through activating endoplasmic reticulum stress in colon cancer cells.
Epibrassinolide (EBR), a member of brassinostreoids plant hormones with cell proliferation promoting role in plants, is a natural polyhydroxysteroid with structural similarity to steroid hormones of vertebrates. EBR has antiproliferative and apoptosis-inducing effect in various cancer cells. Although EBR has been shown to affect survival and mitochondria-mediated apoptosis pathways in a p53-independent manner, the exact molecular targets of EBR are still under investigation. Our recent SILAC (Stable Isotope Labeling by Amino Acids in Cell Culture) data showed that the most significantly altered protein after EBR treatment was calreticulin (CALR). CALR, a chaperone localized in endoplasmic reticulum (ER) lumen, plays role in protein folding and buffering Ca2+ ions. The alteration of CALR may cause ER stress and unfolded protein response correspondingly the induction of apoptosis. Unfolded proteins are conducted to 26S proteasomal degradation following ubiquitination. Our study revealed that EBR treatment caused ER stress and UPR by altering CALR expression causing caspase-dependent apoptosis in HCT 116, HT29, DLD-1, and SW480 colon cancer cells. Furthermore, 48 h EBR treatment did not caused UPR in Fetal Human Colon cells (FHC) and Mouse Embryonic Fibroblast cells (MEF). In addition our findings showed that HCT 116 colon cancer cells lacking Bax and Puma expression still undergo UPR and related apoptosis. CALR silencing and rapamycin co-treatment prevented EBR-induced UPR and apoptosis, whereas 26S proteasome inhibition further increased the effect of EBR in colon cancer cells. All these findings showed that EBR is an ER stress and apoptotic inducer in colon cancer cells without affecting non-malignant cells.


Contact-based ligand-clustering approach for the identification of active compounds in virtual screening.
Evaluation of docking results is one of the most important problems for virtual screening and in silico drug design. Modern approaches for the identification of active compounds in a large data set of docked molecules use energy scoring functions. One of the general and most significant limitations of these methods relates to inaccurate binding energy estimation, which results in false scoring of docked compounds. Automatic analysis of poses using self-organizing maps (AuPosSOM) represents an alternative approach for the evaluation of docking results based on the clustering of compounds by the similarity of their contacts with the receptor. A scoring function was developed for the identification of the active compounds in the AuPosSOM clustered dataset. In addition, the AuPosSOM efficiency for the clustering of compounds and the identification of key contacts considered as important for its activity, were also improved. Benchmark tests for several targets revealed that together with the developed scoring function, AuPosSOM represents a good alternative to the energy-based scoring functions for the evaluation of docking results.


Psychophysical properties of two-stroke apparent motion.
In two-stroke apparent movement, repeated presentation of a two-frame pattern displacement followed by a brief inter-stimulus interval (ISI) can create an impression of continuous forward motion (G. Mather, 2006). Does the ISI in two-stroke motion just break the connection between adjacent frames, switching off the motion signal they normally generate, or does it actually generate a reversed motion signal? Reversed apparent motion in two-frame stimuli separated by a brief ISI has been reported in several previous papers (ISI reversal), which found that the effect is optimal at short, mean-luminance ISIs, and is abolished at scotopic luminances. A series of five experiments compared two-stroke apparent motion with ISI reversal using the same stimulus display. The two effects show the same dependence on ISI duration and luminance and are both abolished at low mean luminance. Results therefore support the conclusion that the ISI in two-stroke apparent motion does contribute a reversed motion signal and constrain theoretical explanations of two-stroke apparent motion.


Full-length RAG1 promotes contact with coding and intersignal sequences in RAG protein complexes bound to recombination signals paired in cis.
The RAG proteins initiate V(D)J recombination by mediating synapsis and cleavage of two different antigen receptor gene segments through interactions with their flanking recombination signal sequences (RSS). The protein-DNA complexes that support this process have mainly been studied using RAG-RSS complexes assembled using oligonucleotide substrates containing a single RSS that are paired in trans to promote synapsis. How closely these complexes model those formed on longer, more physiologically relevant substrates containing RSSs on the same DNA molecule (in cis) remains unclear. To address this issue, we characterized discrete core and full-length RAG protein complexes bound to RSSs paired in cis. We find these complexes support cleavage activity regulated by V(D)J recombination's '12/23 rule' and exhibit plasticity in RSS usage dependent on partner RSS composition. DNA footprinting studies suggest that the RAG proteins in these complexes mediate more extensive contact with sequences flanking the RSS than previously observed, some of which are enhanced by full-length RAG1, and associated with synapsis and efficient RSS cleavage. Finally, we demonstrate that the RAG1 C-terminus facilitates hairpin formation on long DNA substrates, and full-length RAG1 promotes hairpin retention in the post-cleavage RAG complex. These results provide new insights into the mechanism of physiological V(D)J recombination.


A Multi-Site Qualitative Study Examining Pharmacy Student Perspectives on the Opioid Crisis.
Objective. To determine pharmacy students' perspectives regarding opioid use, the opioid crisis, and pharmacy education related to both topics.Methods. Students from each professional year at eight participating schools and colleges of pharmacy were invited to participate in focus groups and answer questions about their experiences with the opioid crisis. Faculty and/or staff moderated the focus groups and audio-recorded responses. Recordings were deidentified, transcribed, and analyzed.Results. One hundred fifty students participated in one of 29 focus groups conducted. Responses were categorized according to themes using consensual qualitative research (CQR) methodology. Sources impacting student views on the crisis included school, personal and work experience, and media. Perspective changes since starting school included increased knowledge and awareness and decreased bias/stigma.Conclusion. Conducting focus groups on the opioid crisis provided pharmacy schools with information on what student pharmacists are learning about the crisis, practices they see, and their recommendations to address the crisis. Pharmacy schools can better prepare students to combat the opioid crisis by providing them with training in opioid counseling, use of naloxone, and how to have difficult conversations with patients.


Deciphering Multiplicity of HIV-1C Infection: Transmission of Closely Related Multiple Viral Lineages.
A single viral variant is transmitted in the majority of HIV infections. However, about 20% of heterosexually transmitted HIV infections are caused by multiple viral variants. Detection of transmitted HIV variants is not trivial, as it involves analysis of multiple viral sequences representing intra-host HIV-1 quasispecies. We distinguish two types of multiple virus transmission in HIV infection: (1) HIV transmission from the same source, and (2) transmission from different sources. Viral sequences representing intra-host quasispecies in a longitudinally sampled cohort of 42 individuals with primary HIV-1C infection in Botswana were generated by single-genome amplification and sequencing and spanned the V1C5 region of HIV-1C env gp120. The Maximum Likelihood phylogeny and distribution of pairwise raw distances were assessed at each sampling time point (n = 217; 42 patients; median 5 (IQR: 4-6) time points per patient, range 2-12 time points per patient). Transmission of multiple viral variants from the same source (likely from the partner with established HIV infection) was found in 9 out of 42 individuals (21%; 95 CI 10-37%). HIV super-infection was identified in 2 patients (5%; 95% CI 1-17%) with an estimated rate of 3.9 per 100 person-years. Transmission of multiple viruses combined with HIV super-infection at a later time point was observed in one individual. Multiple HIV lineages transmitted from the same source produce a monophyletic clade in the inferred phylogenetic tree. Such a clade has transiently distinct sub-clusters in the early stage of HIV infection, and follows a predictable evolutionary pathway. Over time, the gap between initially distinct viral lineages fills in and initially distinct sub-clusters converge. Identification of cases with transmission of multiple viral lineages from the same source needs to be taken into account in cross-sectional estimation of HIV recency in epidemiological and population studies.


The crystal structure of p13suc1, a p34cdc2-interacting cell cycle control protein.
p13suc1 binds to p34cdc2 kinase and is essential for cell cycle progression in eukaryotic cells. The crystal structure of S.pombe p13suc1 has been solved to 2.7 A resolution using data collected at the ESRF source, Grenoble, from both native crystals and crystals of a seleno-methionine derivative. The starting point for structure solution was the determination of the six selenium sites by direct methods. The structure is dominated by a four-stranded beta-sheet, with four further alpha-helical regions. p13suc1 crystallizes as a dimer in the asymmetric unit stabilized by the binding of two zinc ions. A third zinc site stabilizes the higher-order crystal packing. The sites are consistent with a requirement for zinc during crystal growth. A likely site for p13suc1-protein interaction is immediately evident on one face of the p13suc1 surface. This region comprises a group of conserved, exposed aromatic and hydrophobic residues below a flexible negatively charged loop. A conserved positively charged area would also present a notable surface feature in the monomer, but is buried at the dimer interface. p13suc1 is larger than its recently solved human homologue p9CKS2, with the extra polypeptide forming a helical N-terminal extension and a surface loop between alpha-helices 3 and 4. Notably, p13suc1 does not show the unusual beta-strand exchange that creates an intimate p9CKS2 dimer. p13suc1 cannot oligomerize to form a stable hexamer as has been proposed for p9CKS2.


Electron-paramagnetic-resonance studies on nitrogenase of Klebsiella pneumoniae. Evidence for acetylene- and ethylene-nitrogenase transient complexes.
Klebsiella pneumoniae nitrogenase exhibited four new electron-paramagnetic-resonance signals during turnover at 10 degrees C, pH7.4, which were assigned to intermediates present in low concentrations in the steady state. 57Fe-substituted Mo--Fe protein showed that they arose from Fe--S clusters in the Mo--Fe protein of nitrogenase. The new signals are designated: Ic, g values at 4.67, 3.37 and approx. 2.0; VI, g values at 2.125, 2.000 and 2.000; VII, g values at 5.7 and 5.4; VIII, g values at 2.092, 1.974 and 1.933. The sharp axial signal VI arises from a Fe4S4 cluster at the --1 oxidation level. This signal was only detected in the presence of ethylene and provides the first evidence of an enzyme--product complex for nitrogenase. [13C]Acetylene and [13C]ethylene provided no evidence for direct binding of this substrate and product to the Fe--S clusters giving rise to these signals. The dependence of signal intensities on acetylene concentration indicated two types of binding site, with apparent dissociation constants K less than 16 micron and K approximately 13mM. A single binding site for ethylene (K=1.5mM) was detected. A scheme is proposed for the mechanism of reduction of acetylene to ethylene and inhibition of this reaction by CO.


Cell freezing protocol suitable for ATAC-Seq on motor neurons derived from human induced pluripotent stem cells.
In recent years, the assay for transposase-accessible chromatin using sequencing (ATAC-Seq) has become a fundamental tool of epigenomic research. However, it is difficult to perform this technique on frozen samples because freezing cells before extracting nuclei can impair nuclear integrity and alter chromatin structure, especially in fragile cells such as neurons. Our aim was to develop a protocol for freezing neuronal cells that is compatible with ATAC-Seq; we focused on a disease-relevant cell type, namely motor neurons differentiated from induced pluripotent stem cells (iMNs) from a patient affected by spinal muscular atrophy. We found that while flash-frozen iMNs are not suitable for ATAC-Seq, the assay is successful with slow-cooled cryopreserved cells. Using this method, we were able to isolate high quality, intact nuclei, and we verified that epigenetic results from fresh and cryopreserved iMNs quantitatively agree.


A prospective study of in-utero exposure to magnetic fields and the risk of childhood obesity.
We conducted a prospective study to examine whether in-utero exposure to magnetic fields (MFs) increases the risk of childhood obesity. Participating women carried a meter measuring MF levels during pregnancy and 733 of their children were followed up to 13 years to collect clinically recorded information on growth patterns with 33 weight measurements per child on average. Prenatal exposure to high MF level was associated with increased risk of being obese in offspring than those with lower MF level (odds ratio = 1.69, 95% confidence interval: 1.01-2.84). The association demonstrated a dose-response relationship and was stronger (more than 2.3 fold increased risk) among children who were followed up to the end of the study. The association existed only for persistent obesity, but not for transitory (unlikely) obesity. Maternal exposure to high MF during pregnancy may be a new and previously unknown factor contributing to the world-wide epidemic of childhood obesity/overweight.


COVID-19 and Cannabidiol (CBD).
COVID-19 pandemic has resulted in devastating mortality and morbidity consisting of socioeconomic and health effects that have included respiratory/pulmonary, cardiovascular, mental health and neurological consequences such as anxiety, depression, and substance use. Extensive efforts are underway to develop preventive vaccines and therapeutics such as remdesivir, dexamethasone, convalescent plasma, and others to treat COVID-19 but many report residual mental health problems after recovery. Cannabis products such as cannabidiol (CBD) are being advertised for the treatment of COVID-19 associated mental health problems and substance use disorders. This commentary will briefly clear the myth that CBD can ameliorate a wide range of COVID-19 associated health effects including anxiety, depression, or any substance use disorder, and show that there is a clear lack of sufficient unbiased clinical evidence from well-designed double-blind, placebo-controlled clinical trials to prove the antianxiety or antidepression therapeutic properties of CBD and support its wide use as medicine to treat COVID-19- associated mental health conditions or substance use disorders. Finally, we suggest that addiction physicians must play an important role in dealing with their patients requesting CBD prescription for treating any of these conditions.


Primary retroperitoneal hydatid cyst with unusual clinical manifestation.
Although hydatid disease may affect any organ of the body, there are only a limited number of cases where the primary lesion is in the retroperitoneal region. Since the clinical and laboratory findings are nonspecific, the correct preoperative diagnosis may be difficult. A primary retroperitoneal hydatid cyst that was given a preoperative diagnosis of adrenal mass and treated by adrenalectomy and partial nephrectomy is presented and discussed with the relevant literature.


Genomic approaches to bleeding disorders.
The genes encoding the coagulation factors were characterized over two decades ago. Since then, significant progress has been made in the genetic diagnosis of the two commonest severe inherited bleeding disorders, haemophilia A and B. Experience with the genetic of inherited rare bleeding disorders and platelet disorders is less well advanced. Rare bleeding disorders are usually inherited as autosomal recessive disorders, while it is now clear that a number of the more common platelet function disorders are inherited as autosomal dominant traits. In both cases, DNA sequencing has been useful since most of these disorders are due to mutations located at the coding regions or splice sites of genes encoding the abnormal protein. However, in 5-10% of patients affected with severe clotting factor deficiencies, no genetic defect can be identified and until recently, the genetic characterization of inherited platelet disorders had been confined to the more prevalent conditions such as Glanzmann disease and Bernard-Soulier syndrome. In patients with no gene mutations identified, so far, the role of next-generation sequencing as well as of other new genomic technologies will very likely have increasing importance. However, such methods require extensive bioinformatics analysis that, in turn will require critical revision of our current diagnostic infrastructure.


Dual modulation by l-leucovorin and recombinant human interferon alpha 2a of 5-fluorouracil antitumor activity against the human colon carcinoma xenograft Co-4.
We investigated the dual modulation by l-leucovorin (LV) and recombinant human interferon-alpha 2a (IFN-alpha 2a) of 5-fluorouracil (5-FU) antitumor activity against human colon carcinoma cells (Co-4) using a nude mouse system. 5-FU was administered intraperitoneally (IP) at 10 or 90 mg/kg. 5-FU (10 mg/kg) was administered daily for 10 days, and 90 mg/kg was administered once. LV was administered IP 1 and 0 h before 5-FU treatment at 200 mg/kg. IFN-alpha 2a was administered subcutaneously (SC) daily for 14 days at 60,000 IU/mouse. When 5-FU was administered at 10 or 90 mg/kg with these two modulators, the antitumor effect was increased significantly, with T/C ratios of 18.1 and 6.1, respectively. These modulatory effects were assessed as synergistic, without associated severe side effects or death during the experimental period. LV augmented the antitumor activity of 5-FU through increment of thymidylate synthetase (TS) inhibition, and IFN-alpha 2a showed a modulatory effect in elevating the intratumoral concentration of fluorouridine without change in TS inhibition. These results suggest that 5-FU antitumor activity against human colon carcinoma could be significantly potentiated without severe side effects by these two modulators, which possess different modes of action.


Elastic serum-albumin based hydrogels: mechanism of formation and application in cardiac tissue engineering.
Hydrogels are promising materials for mimicking the extra-cellular environment. Here, we present a simple methodology for the formation of a free-standing viscoelastic hydrogel from the abundant and low cost protein serum albumin. We show that the mechanical properties of the hydrogel exhibit a complicated behaviour as a function of the weight fraction of the protein component. We further use X-ray scattering to shed light on the mechanism of gelation from the formation of a fibrillary network at low weight fractions to interconnected aggregates at higher weight fractions. Given the match between our hydrogel elasticity and that of the myocardium, we investigated its potential for supporting cardiac cells in vitro. Interestingly, these hydrogels support the formation of several layers of myocytes and significantly promote the maintenance of a native-like gene expression profile compared to those cultured on glass. When confronted with a multicellular ventricular cell preparation, the hydrogels can support macroscopically contracting cardiac-like tissues with a distinct cell arrangement, and form mm-long vascular-like structures. We envisage that our simple approach for the formation of an elastic substrate from an abundant protein makes the hydrogel a compelling biomedical material candidate for a wide range of cell types.


An integrated approach to improving the availability and utilisation of tuberculosis healthcare in rural South Africa.
Patients with tuberculosis (TB) face several challenges in accessing care, and an integrated service that includes HIV testing could be preferable for them and ensure timely HIV treatment initiation and optimal TB care. To investigate factors, including uptake of the offer of HIV testing, associated with availability and utilisation of healthcare by TB patients in a rural programme devolved to primary care in Hlabisa sub-district, KwaZulu-Natal. Three hundred TB patients were randomly selected in a two-stage-sampling scheme with five primary healthcare clinic (PHC) sampling units selected with probability proportional to size. Data were collected using a structured questionnaire. We describe key availability and utilisation factors and analyse factors associated with being offered an HIV test in multiple regressions controlling for sex, age, education, employment and marital status. Most patients (75.2%) received care for a first episode of TB, mainly pulmonary. Nearly all (94.3%) were offered an HIV test during their current TB treatment episode, patients using their closest clinic being substantially more likely to have been offered HIV testing than those not using their closest clinic (adjusted odds ratio 12.79, p=0.05). About one-fifth (20.3%) of patients did not take medication under observation, and 3.4% reported missing taking their tablets at some stage. Average travelling time to the clinic and back was 2 hours, most patients (56.8%) using minibus taxis. We demonstrate high HIV testing rates among TB patients in a rural public programme, suggesting appropriate management of HIV-TB co-infected patients. We describe healthcare availability and utilisation factors that can inform the proposed district management teams for PHC re-engineering on areas needing improvement.


Structural neuroimaging correlates of social deficits are similar in autism spectrum disorder and attention-deficit/hyperactivity disorder: analysis from the POND Network.
Autism spectrum disorder (ASD), attention-deficit/hyperactivity disorder (ADHD), and obsessive-compulsive disorder (OCD) have been associated with difficulties recognizing and responding to social cues. Neuroimaging studies have begun to map the social brain; however, the specific neural substrates contributing to social deficits in neurodevelopmental disorders remain unclear. Three hundred and twelve children underwent structural magnetic resonance imaging of the brain (controls = 32, OCD = 44, ADHD = 77, ASD = 159; mean age = 11). Their social deficits were quantified on the Social Communication Questionnaire (SCQ) and the Reading the Mind in the Eyes Test (RMET). Multivariable regression models were used to examine the structural neuroimaging correlates of social deficits, with both a region of interest and a whole-brain vertex-wise approach. For the region of interest analysis, social brain regions were grouped into three networks: (1) lateral mentalization (e.g., temporal-parietal junction), (2) frontal cognitive (e.g., orbitofrontal cortex), and (3) subcortical affective (e.g., limbic system) regions. Overall, social communication deficits on the SCQ were associated with thinner cortices in the left lateral regions and the right insula, and decreased volume in the ventral striatum, across diagnostic groups (p = 0.006 to <0.0001). Smaller subcortical volumes were associated with more severe social deficits on the SCQ in ASD and ADHD, and less severe deficits in OCD. On the RMET, larger amygdala/hippocampal volumes were associated with fewer deficits across groups. Overall, patterns of associations were similar in ASD and ADHD, supporting a common underlying biology and the blurring of the diagnostic boundaries between these disorders.


Steviol glycosides enhance pancreatic beta-cell function and taste sensation by potentiation of TRPM5 channel activity.
Steviol glycosides (SGs), such as stevioside and rebaudioside A, are natural, non-caloric sweet-tasting organic molecules, present in extracts of the scrub plant Stevia rebaudiana, which are widely used as sweeteners in consumer foods and beverages. TRPM5 is a Ca2+-activated cation channel expressed in type II taste receptor cells and pancreatic β-cells. Here we show that stevioside, rebaudioside A and their aglycon steviol potentiate the activity of TRPM5. We find that SGs potentiate perception of bitter, sweet and umami taste, and enhance glucose-induced insulin secretion in a Trpm5-dependent manner. Daily consumption of stevioside prevents development of high-fat-diet-induced diabetic hyperglycaemia in wild-type mice, but not in Trpm5-/- mice. These results elucidate a molecular mechanism of action of SGs and identify TRPM5 as a potential target to prevent and treat type 2 diabetes.


Fabry disease: results of the first UK hemodialysis screening study.
Fabry disease is an X-linked lysosomal storage disorder in which deficiency of α-Galactosidase A (α-Gal A), leads to accumulation of glycosphingolipids in the vascular endothelium, kidneys and heart. Males with classical disease present in childhood, however some individuals with low levels of α-Gal A activity present atypically with adult onset renal impairment. Screening studies in patients with established end-stage renal failure (ESRF) suggest that up to 1.5% of patients have sub-normal α-Gal A levels. We used the dried blood spot (DBS) enzyme activity test to screen for undiagnosed Fabry disease in patients with ESRF. Male hemodialysis patients treated at a single UK center (n = 155) were screened using the DBS assay. In patients with low enzyme activity on DBS, α-Gal A activity was assessed in plasma and leucocytes. 8 of the 155 (5%) patients screened showed low enzyme activity on the DBS assay. Confirmatory testing of plasma and leucocyte α-Gal A activity showed normal activity in all cases tested, indicating a false positive DBS result. This study is the first screening program in UK hemodialysis patients using the DBS test and did not identify any new cases of Fabry disease. In this cohort, the DBS enzyme assay had a false positive rate of 2.6%, emphasizing the need for validation with alternative techniques.


Papillary thyroid microcarcinoma: a study of 535 cases observed in a 50-year period.
The study aims were to characterize patients with papillary thyroid microcarcinoma and to provide data on outcome after surgical therapy. Five hundred thirty-five patients with papillary microcarcinoma had initial treatment at Mayo Clinic from 1940 to 1989. Follow-up extended to 48 years. Median follow-up time for 400 survivors was 16 years. Recurrence and mortality details were derived from a computerized cancer database. Median tumor size was 8 mm. Ninety-nine percent of tumors were histologic grade 1; 98% were not locally invasive. Thirty-two percent of patients had nodal metastases at examination. TNM stages were I in 485 patients (91%), III in 49 patients (9%), and IV in one patient (0.2%). Ninety-one percent of patients underwent bilateral lobar resection. Tumor resection was incomplete in three cases (0.6%). Radioiodine remnant ablation was performed in 55 patients (10%). All-causes survival did not differ from expected; two patients (0.4%) died of papillary microcarcinoma. Twenty-year tumor recurrence rate was 6%. Higher recurrence rates were seen either with node-positive patients (p < 0.0001) or after unilateral lobectomy (p < 0.0001). Recurrence rates did not appear to be significantly altered by total thyroidectomy (p = 0.44) or radioiodine remnant ablation in node-positive patients (p = 0.99). These results reaffirm that papillary microcarcinoma has an excellent prognosis if managed initially by bilateral lobar resection. Routine radioiodine remnant ablation is not indicated.


The Receptor for Activated C Kinase in Plant Signaling: Tale of a Promiscuous Little Molecule.
Two decades after the first report of the plant homolog of the Receptor for Activated C Kinase 1 (RACK1) in cultured tobacco BY2 cells, a significant advancement has been made in the elucidation of its cellular and molecular role. The protein is now implicated in many biological functions including protein translation, multiple hormonal responses, developmental processes, pathogen infection resistance, environmental stress responses, and miRNA production. Such multiple functional roles are consistent with the scaffolding nature of the plant RACK1 protein. A significant advance was achieved when the β-propeller structure of the Arabidopsis RACK1A isoform was elucidated, thus revealing that its conserved seven WD repeats also assembled into this typical topology. From its crystal structure, it became apparent that it shares the structural platform for the interaction with ligands identified in other systems such as mammals. Although RACK1 proteins maintain conserved Protein Kinase C binding sites, the lack of a bona fide PKC adds complexity and enigma to the nature of the ligand partners with which RACK1 interacts in plants. Nevertheless, ligands recently identified using the split-ubiquitin based and conventional yeast two-hybrid assays, have revealed that plant RACK1 is involved in several processes that include defense response, drought and salt stress, ribosomal function, cell wall biogenesis, and photosynthesis. The information acquired indicates that, in spite of the high degree of conservation of its structure, the functions of the plant RACK1 homolog appear to be distinct and diverse from those in yeast, mammals, insects, etc. In this review, we take a critical look at the novel information regarding the many functions in which plant RACK1 has been reported to participate, with a special emphasis on the information on its currently identified and missing ligand partners.


Effects of fibrous cap thickness on peak circumferential stress in model atherosclerotic vessels.
It is likely that factors other than stenosis severity predispose some atherosclerotic plaques to rupture. Because focal increases in circumferential stress may be an important mechanism of plaque rupture, we examined peak circumferential stress of atherosclerotic lesions by using finite element analysis based on idealized two-dimensional cross sections of diseased vessels similar to intravascular ultrasound images. The study was designed to test the hypothesis that subintimal plaque structural features such as thickness of the fibrous cap are more important factors in the distribution of stress in the plaque than stenosis severity. The analysis incorporated equilibrium biomechanical parameters from normal and diseased vessels and determined the stress distribution within the plaque at a mean luminal internal pressure of 110 mm Hg. With a constant luminal area reduction of 70%, maximum circumferential stress (sigma max) normalized to luminal pressure (sigma max/P) increased from 6.0 to 24.8 as the thickness of the lipid pool was increased from 38% to 54% of the plaque thickness because of the thinner fibrous cap over the lipid pool. When the lipid pool thickness was constant, increasing the stenosis severity from 70% to 91% by increasing the fibrous cap thickness decreased sigma max/P from 24.8 to 4.7. When no lipid pool was present and the stenosis severity was increased from 70% to 99%, sigma max/P decreased from 5.3 to 4.7. Thus, reducing the fibrous cap thickness dramatically increases peak circumferential stress in the plaque, whereas increasing the stenosis severity actually decreases peak stress in the plaque.(ABSTRACT TRUNCATED AT 250 WORDS)


Participation in community-originated interventions is associated with positive changes in weight status and health behaviors in youth.
To assess changes in children's health behaviors and weight status after participation in community-originated interventions. Prospective cohort study following body mass index (BMI) z-score trajectory over time. Schools and community settings in 19 locations in North Carolina. A total of 1144 children, with an average age of 9.5 years, participating in community-originated physical activity and nutrition interventions, such as active recess and healthy cafeteria offerings, and environment and policy changes designed to prevent and reduce the prevalence of childhood obesity. Retention from baseline to final collection was 54%. Measures . Self-reported physical activity and eating behaviors; measured height and weight were collected at baseline and after interventions (average of 20 months). Z-score was compared with expected growth without intervention. Descriptive statistics, χ(2) tests and t-tests, and ANOVA analyses of variance to assess changes in health behaviors and weight status. More than 11% of children improved their weight status, and 86% of children who began at a healthy weight remained there. The average BMI for children who were overweight at enrollment was significantly lower after intervention (1.1 kg/m(2) lower for boys and 0.88 kg/m(2) lower for girls). Overweight and obese children who made improvements in key health behaviors showed greater decreases in BMI z-score than those who did not improve those behaviors (increased fruit [-.2207 vs. -.0793] and vegetable [-.2215 vs. -.0855] intake, and drinking less soda [-.1985 vs. -.0912]). There were no significant increases in physical activity, and changes in physical activity were not related to decreases in BMI z-score in this study. Community-originated interventions to raise awareness about food choices and to change policies and environments may improve BMI z-scores.


Computer-assisted trauma surgery.
Computer-assisted orthopaedic surgery (CAOS) is performed by digitizing the patient's anatomy, combining the images in a computerized system, and integrating the surgical instruments into the digitized image background. This allows the surgeon to navigate the surgical instruments and the bone in an improved, virtual visual environment. CAOS in traumatology is performed with images obtained by fluoroscopy, CT, or three-dimensional fluoroscopy. CAOS is used in basic trauma procedures for preoperative planning, fracture reduction, intramedullary nailing, percutaneous screw or plate fixation, and hardware or shrapnel removal. Potential benefits of CAOS include minimal invasiveness, increased accuracy, and decreased radiation exposure. Limitations include a significant learning curve, increased surgical time, requirements for special setup and equipment handling in the operating room, specialized technical support, and cost. Current evidence shows no advantage with CAOS in trauma cases compared with conventional methods. Prospective randomized trials and clinical outcomes are lacking.


GC-FID and HPLC-DAD Methods for the Determination of Menadione Sodium Bisulphite Directly and by Converting Menadione Sodium Bisulphite to Menadione in Pharmaceutical Preparation.
was performed in both direct analysis of MSB and analysis of MN by converting MSB to MN with sodium carbonate. GC-FID method was carried out on the HP-5 capillary column GC-FID and HPLC-DAD methods were developed for determination of menadione (MN) and menadione sodium bisulphite (MSB). By means of each method, quantitative analysis of MSB in commercial pharmaceutical using nitrogen gas. HPLC-DAD method was achieved on the reversed phase C8 column by using a mobile phase consisting methanol and water. The calibration curves of GC-FID and HPLC-DAD for both analytes were linear in the same concentration range (0.5-20 μg/mL). Both methods were validated in terms of precision, accuracy, recovery and limits of detection (LOD) and quantitation (LOQ). Although LOD values of HPLC-DAD method (0.010 μg/mL for MN and 0.005 μg/mL for MSB) is lower than obtained values with GC-FID method (0.04 μg/mL for MN and 0.06 μg/mL for MSB), both methods gave similar and favorable results in terms of precision and accuracy. The Student's t-test was applied to investigate the significant of the different between the results of MSB determination with direct analysis of MSB and analysis of MN by converting MSB to MN by means of GC-FID and HPLC-DAD method in dosage form.


Effects of nonionic surfactants on the cell surface hydrophobicity and apparent hamaker constant of a Sphingomonas sp.
Nonionic surfactants of the form CxEy were studied for their ability to alter the cell surface hydrophobicity and apparent Hamaker constants of a Sphingomonas sp. Through contact angle measurements on hydrated and dried bacterial lawns, it was found that the cell surface hydrophobicity changed systematically with both the alkyl (x) and polyoxyethylene (y) chain lengths. While differences in contact angles were observed between hydrated and dried lawns, they could not be attributed to the mere presence or absence of water, suggesting that reorientation of cell surface components may occur during drying. All surfactants examined reduced the hydrophilicity of the bacterial cell surface, with one surfactant (C18E10) making the cells hydrophobic. Effective bacterial Hamaker constants for bacteria interacting across a vacuum (Abb) and water (Abwb) and bacteria interacting in water with quartz sand (Abws) were calculated from the contact angles. It was found that the surfactants have the potential to reduce the Hamaker constants, but that the overall effects differed between dried and hydrated lawns, indicating that lawn preparation method can have a significant impact in interpretation of cell surface properties. The results also indicate that the Abws value of 10-20 J, which is often assumed in bacterial attachment and transport studies, may be an order of magnitude higher than the actual value. Finally, the results suggest that alteration of bacterial adhesion due to the presence of surfactants cannot be attributed to a single cell surface property but is rather due to multiple interactions.


Simultaneous enantiomeric determination of propranolol, metoprolol, pindolol, and atenolol in natural waters by HPLC on new polysaccharide-based stationary phase using a highly selective molecularly imprinted polymer extraction.
A simple high performance liquid chromatography method HPLC-UV for simultaneous enantiomeric determination of propranolol, metoprolol, pindolol, and atenolol in natural water samples was developed and validated, using a molecularly imprinted polymer solid-phase extraction. To achieve this purpose, Lux(®) Cellulose-1/Sepapak-1 (cellulose tris-(3,5-dymethylphenylcarbamate)) (Phenomenex, Madrid, Spain) chiral stationary phase was used in gradient elution and normal phase mode at ambient temperature. The gradient elution program optimized consisted of a progressive change of the mobile phase polarity from n-hex/EtOH/DEA 90/10/0.5 (v/v/v) to 60/40/0.5 (v/v/v) in 13 min, delivered at a flow rate of 1.3 ml/min and a sudden change of flow rate to 2.3 ml/min in 1 min. Critical steps in any molecularly imprinted polymer extraction protocol such as the flow rate to load the water sample in the cartridges and the breakthrough volume were optimized to obtain the higher extraction recoveries for all compounds. In optimal conditions (100 ml breakthrough volume loaded at 2.0 ml/min), extraction recoveries for the four pairs of β-blockers were near 100%. The MIP-SPE-HPLC-UV method developed demonstrates good linearity (R(2) ≥ 0.99), precision, selectivity, and sensitivity. Method limit detection was 3.0 µg/l for propranolol and pindolol enantiomers and 20.0 and 22.0 µg/l for metoprolol and atenolol enantiomers, respectively. The proposed methodology should be suitable for routine control of these emerging pollutants in natural waters for a better understanding of the environmental impact and fate.


Hosts use altered macronutrient intake to circumvent parasite-induced reduction in fecundity.
Explanations for the evolution of pathogen-induced fecundity reduction usually rely on a common principle: the trade-off between host longevity and reproduction. Recent advances in nutritional research have, however, challenged this assumption and shown that longevity and reproduction are not inextricably linked. In this study, we showed that beetles infected by cysticercoids of the tapeworm Hymenolepis diminuta increased their total food intake and, more particularly, their carbohydrate consumption compared with uninfected insects. This increased intake was only pronounced during the first 12 days p.i., when the parasite grows and develops into a mature metacestode. Despite consuming more nutrients, infected individuals sustained lower levels of body lipid and were less efficient at converting ingested protein to body protein. However they demonstrated a capacity to compose a diet that sustained high levels of reproductive output unless confined to foods that were nutritionally dilute. We did not find any indication that macronutrient intakes had an effect on host pro-phenoloxidase activity; however, phenoloxidase activity was significantly affected by protein intake. Our results showed that when offered nutritionally complementary diets, infected hosts do not systematically suffer a reduction in fecundity. Thus, in our view, the assumption that a reduction in host reproduction represents an adaptive response by the host or the parasite to divert resources away from reproduction toward other traits should be reassessed.


Cross-ecosystem nutrient subsidies in Arctic and alpine lakes: implications of global change for remote lakes.
Environmental change is continuing to affect the flow of nutrients, material and organisms across ecosystem boundaries. These cross-system flows are termed ecosystem subsidies. Here, we synthesize current knowledge of cross-ecosystem nutrient subsidies between remote lakes and their surrounding terrain, cryosphere, and atmosphere. Remote Arctic and alpine lakes are ideal systems to study the effects of cross ecosystem subsidies because (a) they are positioned in locations experiencing rapid environmental changes, (b) they are ecologically sensitive to even small subsidy changes, (c) they have easily defined ecosystem boundaries, and (d) a variety of standard methods exist that allow for quantification of lake subsidies and their impacts on ecological communities and ecosystem functions. We highlight similarities and differences between Arctic and alpine systems and identify current knowledge gaps to be addressed with future work. It is important to understand the dynamics of nutrient and material flows between lakes and their environments in order to improve our ability to predict ecosystem responses to continued environmental change.


Nef-induced CD4 degradation: a diacidic-based motif in Nef functions as a lysosomal targeting signal through the binding of beta-COP in endosomes.
The Nef protein of primate lentiviruses downregulates the cell surface expression of CD4 through a two-step process. First, Nef connects the cytoplasmic tail of CD4 with adaptor protein complexes (AP), thereby inducing the formation of CD4-specific clathrin-coated pits that rapidly endocytose the viral receptor. Second, Nef targets internalized CD4 molecules for degradation. Here we show that Nef accomplishes this second task by acting as a connector between CD4 and the beta subunit of COPI coatomers in endosomes. A sequence encompassing a critical acidic dipeptide, located nearby but distinct from the AP-binding determinant of HIV-1 Nef, is responsible for beta-COP recruitment and for routing to lysosomes. A novel class of endosomal sorting motif, based on acidic residues, is thus revealed, and beta-COP is identified as its downstream partner.


Simultaneous determination of perfluoroalkyl substances and bile acids in human serum using ultra-high-performance liquid chromatography-tandem mass spectrometry.
There is evidence of a positive association between per- and polyfluoroalkyl substances (PFASs) and cholesterol levels in human plasma, which may be due to common reabsorption of PFASs and bile acids (BAs) in the gut. Here we report development and validation of a method that allows simultaneous, quantitative determination of PFASs and BAs in plasma, using 150 μL or 20 μL of sample. The method involves protein precipitation using 96-well plates. The instrumental analysis was performed with ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS), using reverse-phase chromatography, with the ion source operated in negative electrospray mode. The mass spectrometry analysis was carried out using multiple reaction monitoring mode. The method proved to be sensitive, robust, and with sufficient linear range to allow reliable determination of both PFASs and BAs. The method detection limits were between 0.01 and 0.06 ng mL-1 for PFASs and between 0.002 and 0.152 ng mL-1 for BAs, with the exception of glycochenodeoxycholic acid (0.56 ng mL-1). The PFAS measured showed excellent agreement with certified plasma PFAS concentrations in NIST SRM 1957 reference serum. The method was tested on serum samples from 20 healthy individuals. In this proof-of-concept study, we identified significant associations between plasma PFAS and BA levels, which suggests that PFAS may alter the synthesis and/or uptake of BAs. Graphical Abstract.


Exciting Andreev pairs in a superconducting atomic contact.
The Josephson effect describes the flow of supercurrent in a weak link-such as a tunnel junction, nanowire or molecule-between two superconductors. It is the basis for a variety of circuits and devices, with applications ranging from medicine to quantum information. Experiments using Josephson circuits that behave like artificial atoms are now revolutionizing the way we probe and exploit the laws of quantum physics. Microscopically, the supercurrent is carried by Andreev pair states, which are localized at the weak link. These states come in doublets and have energies inside the superconducting gap. Existing Josephson circuits are based on properties of just the ground state of each doublet, and so far the excited states have not been directly detected. Here we establish their existence through spectroscopic measurements of superconducting atomic contacts. The spectra, which depend on the atomic configuration and on the phase difference between the superconductors, are in complete agreement with theory. Andreev doublets could be exploited to encode information in novel types of superconducting qubits.


Comparison of coronary angiography characteristics among acute coronary syndrome patients in young and old age patients at Cipto Mangunkusumo Hospital Jakarta.
to identify the difference between coronary angiography in patients with acute coronary syndrome (ACS) aged 45 years and those aged >45 years. a total of 322 ACS patients who underwent coronary angiography in ICCU of Cipto Mangunkusumo Hospital, Jakarta between January 2008 and December 2012 were included in this study. The severity of coronary stenosis was determined by vessel score and coronary score. A significant vessel score was defined as a stenosis of coronary vessel of >70%. Patients were divided into two groups: those aged 45 years (72 cases) and those aged >45 years (250 cases). The statistical analysis was performed with Chi-square test for categorical data with two-unpaired groups and the t-test or Mann-Whitney for numerical data. the highest distribution of 1-VD (single-vessel disease) patients was found the group of patients aged 45 years (43.1% vs 26.0%); while for 3-VD (triple-vessel disease) patients, the highest distribution was noticed in the group of patients aged >45 years (31.6% vs 18.1%). The stenosis score was lower in patients aged 45 years compared those aged >45 years (median stenosis score 4 vs 8), p<0.001. Atherosclerosis was found most common for the left anterior descending artery in both age groups (65.3% and 74.0%). However, there was less significant stenosis lesion for the Left Circumflex and Right Coronary Arteries in patients aged 45. The result was statistically significant (26.4% and 31.9% vs 46.4% and 57.2%, p=0.002 and 0.001). the number of coronary artery diseases (vessel score) and stenosis score is lower in patients aged 45 years compared to patients aged >45 years.


Real-time, interactive website for US-county level Covid-19 event risk assessment.
Large events and gatherings, particularly those taking place indoors, have been linked to multi-transmission events that have accelerated the pandemic spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). To provide real-time, geo-localized risk information, we developed an interactive online dashboard that estimates the risk that at least one individual with SARS-CoV-2 is present in gatherings of different sizes in the United States. The website combines documented case reports at the county level with ascertainment bias information obtained via population-wide serological surveys to estimate real time circulating, per-capita infection rates. These rates are updated daily as a means to visualize the risk associated with gatherings, including county maps and state-level plots. The website provides data-driven information to help individuals and policy-makers make prudent decisions (e.g., increasing mask wearing compliance and avoiding larger gatherings) that could help control the spread of SARS-CoV-2, particularly in hard-hit regions.


Personality characteristics of adult survivors of childhood trauma.
Lenore Terr (1991) proposed a framework for childhood trauma that distinguishes between single-incident trauma (Type I) and repeated or prolonged trauma (Type II). Terr's framework and data collected from the National Comorbidity Survey (NCS) are used to examine differences in personality between adult survivors of childhood Type I trauma, childhood Type II trauma, and a control group of adults not traumatized as children. Groups were compared on five personality scales. Results indicate that individuals in the trauma categories scored higher in neuroticism and openness to new experiences than individuals in the control group. The Type I group was lower than the control group on interpersonal dependency, whereas the Type II group scored higher than the control group on this measure.


The role of pdx1 and HNF6 in proliferation and differentiation of endocrine precursors.
Ex vivo expansion of embryonic stem cells (ES cells) or pancreatic stem cells for insulin delivery to diabetic patients provides potential for the restoration of islet function in these individuals. Understanding the spatial and temporal requirements of crucial factors for endocrine progenitor specification, proliferation, and terminal differentiation remains a major challenge in the field of pancreas development. Here, we provide speculation as to the role of pdx1 and HNF6 in these different stages of pancreatic endocrine cell development. At the time when islets begin to form within the pancreas, the expression patterns of pdx1 and HNF6 diverge, suggesting distinct functions for each of the genes over the course of endocrine cell development. The current body of evidence provides support for a role of both factors in early endocrine specification as well as a requirement for pdx1 in the generation of mature pancreatic endocrine cells. The precise temporal requirement of HNF6 in the production of terminally differentiated endocrine cells remains unclear. Future studies in this area will rely on conditionally manipulatable systems in combination with lineage-tracing studies for a more accurate assessment of pdx1 and HNF6 function at different stages along the pathway of endocrine cell development.


Adverse outcome analyses of observational data: assessing cardiovascular risk in HIV disease.
Clinical decisions are ideally based on randomized trials but must often rely on observational data analyses, which are less straightforward and more influenced by methodology. The authors, from a series of expert roundtables convened by the Forum for Collaborative HIV Research on the use of observational studies to assess cardiovascular disease risk in human immunodeficiency virus infection, recommend that clinicians who review or interpret epidemiological publications consider 7 key statistical issues: (1) clear explanation of confounding and adjustment; (2) handling and impact of missing data; (3) consistency and clinical relevance of outcome measurements and covariate risk factors; (4) multivariate modeling techniques including time-dependent variables; (5) how multiple testing is addressed; (6) distinction between statistical and clinical significance; and (7) need for confirmation from independent databases. Recommendations to permit better understanding of potential methodological limitations include both responsible public access to de-identified source data, where permitted, and exploration of novel statistical methods.


Selective deletion of PPARβ/δ in fibroblasts causes dermal fibrosis by attenuated LRG1 expression.
Connective tissue diseases of the skin are characterized by excessive collagen deposition in the skin and internal organs. Fibroblasts play a pivotal role in the clinical presentation of these conditions. Nuclear receptor peroxisome-proliferator activated receptors (PPARs) are therapeutic targets for dermal fibrosis, but the contribution of the different PPAR subtypes are poorly understood. Particularly, the role of fibroblast PPARβ/δ in dermal fibrosis has not been elucidated. Thus, we generated a mouse strain with selective deletion of PPARβ/δ in the fibroblast (FSPCre-Pparb/d-/-) and interrogated its epidermal and dermal transcriptome profiles. We uncovered a downregulated gene, leucine-rich alpha-2-glycoprotein-1 (Lrg1), of previously unknown function in skin development and architecture. Our findings suggest that the regulation of Lrg1 by PPARβ/δ in fibroblasts is an important signaling conduit integrating PPARβ/δ and TGFβ1-signaling networks in skin health and disease. Thus, the FSPCre-Pparb/d-/- mouse model could serve as a novel tool in the current gunnery of animal models to better understand dermal fibrosis.


The Ser/Thr Kinase PrkC Participates in Cell Wall Homeostasis and Antimicrobial Resistance in Clostridium difficile.
Clostridium difficile is the leading cause of antibiotic-associated diarrhea in adults. During infection, C. difficile must detect the host environment and induce an appropriate survival strategy. Signal transduction networks involving serine/threonine kinases (STKs) play key roles in adaptation, as they regulate numerous physiological processes. PrkC of C. difficile is an STK with two PASTA domains. We showed that PrkC is membrane associated and is found at the septum. We observed that deletion of prkC affects cell morphology with an increase in mean size, cell length heterogeneity, and presence of abnormal septa. A ΔprkC mutant was able to sporulate and germinate but was less motile and formed more biofilm than the wild-type strain. Moreover, a ΔprkC mutant was more sensitive to antimicrobial compounds that target the cell envelope, such as the secondary bile salt deoxycholate, cephalosporins, cationic antimicrobial peptides, and lysozyme. This increased susceptibility was not associated with differences in peptidoglycan or polysaccharide II composition. However, the ΔprkC mutant had less peptidoglycan and released more polysaccharide II into the supernatant. A proteomic analysis showed that the majority of C. difficile proteins associated with the cell wall were less abundant in the ΔprkC mutant than the wild-type strain. Finally, in a hamster model of infection, the ΔprkC mutant had a colonization delay that did not significantly affect overall virulence.


Variations in reverse transcriptase and RNase H domain mutations in human immunodeficiency virus type 1 clinical isolates are associated with divergent phenotypic resistance to zidovudine.
Mutations in the RNase H domain of human immunodeficiency virus type 1 RT have been reported to cause resistance to zidovudine (ZDV) in vitro. However, very limited data on the in vivo relevance of these mutations in patients exist to date. This study was designed to determine the relationship between mutations in the RNase H domain and viral susceptibility to nucleoside analogues. Viruses harboring complex thymidine analogue mutation (TAM) and nucleoside analogue mutation (NAM) profiles were evaluated for their phenotypic susceptibilities to ZDV, tenofovir (TNF), and the nonapproved nucleoside reverse transcriptase inhibitors (NRTIs) beta-2',3'-didehydro-2',3'-dideoxy-5-fluorocytidine (Reverset), beta-D-5-fluorodioxolane-cytosine, and apricitabine. As controls, viruses from NRTI-naïve patients were also studied. The pol RT region (codons 21 to 250) of the viruses were sequenced and evaluated for mutations in the RNase H domain (codons 441 to 560) and the connection domain (codons 289 to 400). The results showed that viruses from patients failing multiple NRTI-containing regimens had distinct TAM and NAM profiles that conferred various degrees of resistance to ZDV (0.9- to >300-fold). Sequencing of the RNase H domain identified five positions (positions 460,468, 483, 512, and 519) at which extensive amino acid polymorphisms common in both wild-type viruses and viruses from treated patients were identified. No mutations were observed at positions 539 and 549, which have previously been associated with ZDV resistance. Mutations in the RNase H domain did not appear to correlate with the levels of phenotypic resistance to ZDV. Although some mutations were also observed in the connection domain, the simultaneous presence of the L74V and M184V mutations was the most significant determinant of phenotypic resistance to ZDV in patients infected with viruses with TAMs.


Couples' daily self-regulation: The Health Action Process Approach at the dyadic level.
Regulating health behavior change often occurs in a dyadic context of romantic relationships. Dyadic approaches to standard health behavior change models are, however, barely considered. We investigated volitional processes of the Health Action Process Approach model for two health behaviors within a dyadic context of romantic couples. Specifically, we tested whether day-to-day volitional self-regulation predicted one's own and one's partner's cigarettes smoked (Study 1) and physical activity (Study 2). In two dyadic intensive longitudinal studies (Study 1: 83 dual-smoker couples intending to jointly quit smoking; Study 2: 61 overweight couples intending to become physically active), heterosexual partners independently reported on intention, self-efficacy, action planning, and action control in end-of-day diaries. In Study 1, daily number of cigarettes smoked was assessed via self-report. In Study 2, daily moderate-to-vigorous physical activity (MVPA) was assessed objectively via accelerometers. In both studies, dyadic cross-lagged intensive longitudinal analyses based on the Actor-Partner Interdependence Model were applied. Across both studies, individual's own volitional self-regulation positively predicted one's own health behavior (less cigarettes smoked and more MVPA). One's partner's action control and intention also positively predicted one's own health behavior. A marginal partner effect for self-efficacy was found in the context of smoking only. Behavioral self-regulation is not only relevant for individuals themselves, but some volitional processes may spill over to their partners. This highlights the need to specify couple-level processes involved in health behavior change, and to consider a social context of self-regulation.


Terrestrial polarization imagery obtained from the Space Shuttle: characterization and interpretation.
An experiment to measure the polarization of land, sea, haze, and cloud areas from space was carried aboard the Space Shuttle in Sept. 1985. Digitized polarimetric and photometric imagery in mutually perpendicular planes was derived in the red, green, and blue spectral regions from photographs taken with two synchronized Hasselblad cameras using type 5036 Ektachrome film. Digitization at the NASA Houston Video Digital Analysis Systems Laboratory permitted reduction of the imagery into equipolarimetric contours with a relative accuracy of +/-20% for comparison to ground truth. The Island of Hawaii and adjacent sea and cloud areas were the objects of the specific imagery analyzed. Results show that cloud development is uniquely characterized using percent polarization without requiring precision photometric calibration. Furthermore, sea state and wind direction over the sea could be inferred as well as terrestrial soil texture.


Management of generalized anxiety disorder in primary care: identifying the challenges and unmet needs.
Generalized anxiety disorder (GAD) is one of the most common psychiatric disorders in primary care, although it is often underrecognized and undertreated. GAD is chronic, disabling, and associated with other health problems. Treatment response is often unsatisfactory, but the clinical evidence base for new treatments has expanded substantially in the past decade and suggests a growing range of options for reducing the burden of GAD. The objective of this article was to review current literature on GAD and its management to provide an overview of the clinical importance of GAD in primary care and available treatments. Recent studies (ie, over the past decade) on the epidemiology and treatment of GAD were identified by searching Medline using the term generalized anxiety disorder only and in combination with the terms epidemiology and treatment and for each drug class (benzodiazepines, azapirones, antidepressants, antihistamines, alpha-2-delta ligands, and antipsychotics) and for named drugs (buspirone, venlafaxine, duloxetine, fluoxetine, escitalopram, olanzapine, paroxetine, pregabalin, quetiapine, and risperidone in addition to psychological therapies and cognitive-behavioral therapy. The literature search was conducted in August 2008 for the period 1987-2009. Studies were included if judged to be relevant to a review of the epidemiology and management of GAD. Articles were excluded if they were not written in English or were published more than 10 years before the literature search was conducted. A few older studies were included for which more recent research evidence was not available. Recent national and international guidelines for the management of GAD were also reviewed. Most currently available interventions have similar overall efficacy, and treatment choices should reflect the situation of individual patients. Important unmet needs exist for treatments (1) that work rapidly, with (2) broad spectrum benefits, (3) that can improve rates of remission and well-being, (4) are devoid of risk for withdrawal symptoms, and (5) have few if any adverse interactions with other drugs. Additional needs include (6) safer drugs for the elderly, (7) safe and effective drugs for children with GAD, (8) further evaluation of psychotherapy, and (9) understanding the appropriate circumstances for, and optimal choices of, drug combination. While the development of novel treatments evolves, current management approaches can focus on improving identification and defining optimal use of available therapies for GAD.


Magnetic forces and magnetized biomaterials provide dynamic flux information during bone regeneration.
The fascinating prospect to direct tissue regeneration by magnetic activation has been recently explored. In this study we investigate the possibility to boost bone regeneration in an experimental defect in rabbit femoral condyle by combining static magnetic fields and magnetic biomaterials. NdFeB permanent magnets are implanted close to biomimetic collagen/hydroxyapatite resorbable scaffolds magnetized according to two different protocols . Permanent magnet only or non-magnetic scaffolds are used as controls. Bone tissue regeneration is evaluated at 12 weeks from surgery from a histological, histomorphometric and biomechanical point of view. The reorganization of the magnetized collagen fibers under the effect of the static magnetic field generated by the permanent magnet produces a highly-peculiar bone pattern, with highly-interconnected trabeculae orthogonally oriented with respect to the magnetic field lines. In contrast, only partial defect healing is achieved within the control groups. We ascribe the peculiar bone regeneration to the transfer of micro-environmental information, mediated by collagen fibrils magnetized by magnetic nanoparticles, under the effect of the static magnetic field. These results open new perspectives on the possibility to improve implant fixation and control the morphology and maturity of regenerated bone providing "in site" forces by synergically combining static magnetic fields and biomaterials.


Dietary cholesterol and egg intake in relation to incident cardiovascular disease and all-cause and cause-specific mortality in postmenopausal women.
The potential cardiovascular impact of dietary cholesterol intake has been actively debated for decades. We aimed to evaluate associations of dietary cholesterol and egg intakes with incident cardiovascular disease (CVD) and all-cause and cause-specific mortality. We included 96,831 US postmenopausal women aged 50-79 y without known CVD or cancer during baseline enrollment (1993-1998) of the Women's Health Initiative. Dietary information was collected using a validated FFQ. Incident CVD [i.e., ischemic heart disease (IHD) and stroke] and all-cause and cause-specific mortality were ascertained and adjudicated through February 2018. A total of 9808 incident CVD cases and 19,508 all-cause deaths occurred during a median follow-up of 17.8 y and 18.9 y, respectively. After multivariable adjustment for traditional risk factors and key dietary nutrients including dietary saturated fat, there were modest associations of dietary cholesterol intake with incident CVD (HRQ5versusQ1: 1.12; 95% CI: 1.03, 1.21; P-trend < 0.001) and all-cause mortality (HRQ5versusQ1: 1.09; 95% CI: 1.02, 1.15; P-trend < 0.001). Significant positive associations were also observed between dietary cholesterol and incident IHD (P-trend = 0.007), incident ischemic stroke (P-trend = 0.002), and CVD mortality (P-trend = 0.002), whereas there was an inverse association for incident hemorrhagic stroke (P-trend = 0.037) and no association for mortality from cancer, Alzheimer disease/dementia, respiratory diseases, or other causes (P-trend > 0.05). Higher egg consumption was also associated with modestly higher risk of incident CVD (P-trend = 0.004) and all-cause mortality (P-trend < 0.001), with HRs of 1.14 (95% CI: 1.04, 1.25) and 1.14 (95% CI: 1.07, 1.22), respectively, when comparing ≥1 egg/d with <1 egg/wk. Both higher dietary cholesterol intake and higher egg consumption appeared to be associated with modestly elevated risk of incident CVD and all-cause mortality in US postmenopausal women.