Evaluation of safety and tolerance of microencapsulated Lactobacillus reuteri NCIMB 30242 in a yogurt formulation: a randomized, placebo-controlled, double-blind study.
Probiotic organisms have shown promise in treating diseases. Previously, we have reported on the efficacy of microencapsulated Lactobacillus reuteri NCIMB 30242 in a yogurt formulation at lowering serum cholesterol levels in otherwise healthy hypercholesterolemic adults. This study investigates the safety and toxicology of oral ingestion of microencapsulated L. reuteri NCIMB 30242 in a yogurt formulation. A randomized group of 120 subjects received a dose of 5 × 10(10) CFU microencapsulated L. reuteri NCIMB 30242 in yogurt (n=59) or placebo yogurt (n=61) twice/day for 6 weeks. Clinical chemistry and hematological parameters of safety were analyzed. Fecal samples were collected at these time points for the analysis of deconjugated bile acids. The frequency, duration and intensity of adverse events (AEs) and clinical significance of safety parameters were recorded for both groups. No clinically significant differences between the probiotic yogurt and placebo yogurt treated groups were detected in either the blood clinical chemistry or hematology results and there was no significant increase in fecal deconjugated bile acids (P>0.05) between treated and control groups. The frequency and intensity of AEs was similar in the two groups. These results demonstrate the safe use of this formulation in food.


Altered Glycosylation Patterns Increase Immunogenicity of a Subunit Hepatitis C Virus Vaccine, Inducing Neutralizing Antibodies Which Confer Protection in Mice.
Hepatitis C virus (HCV) infection is a global health problem for which no vaccine is available. HCV has a highly heterogeneous RNA genome and can be classified into seven genotypes. Due to the high genetic and resultant antigenic variation among the genotypes, inducing antibodies capable of neutralizing most of the HCV genotypes by experimental vaccination has been challenging. Previous efforts focused on priming humoral immune responses with recombinant HCV envelope E2 protein produced in mammalian cells. Here, we report that a soluble form of HCV E2 (sE2) produced in insect cells possesses different glycosylation patterns and is more immunogenic, as evidenced by the induction of higher titers of broadly neutralizing antibodies (bNAbs) against cell culture-derived HCV (HCVcc) harboring structural proteins from a diverse array of HCV genotypes. We affirm that continuous and discontinuous epitopes of well-characterized bNAbs are conserved, suggesting that sE2 produced in insect cells is properly folded. In a genetically humanized mouse model, active immunization with sE2 efficiently protected against challenge with a heterologous HCV genotype. These data not only demonstrate that sE2 is a promising HCV vaccine candidate, but also highlight the importance of glycosylation patterns in developing subunit viral vaccines. A prophylactic vaccine with high efficacy and low cost is urgently needed for global control of HCV infection. Induction of broadly neutralizing antibodies against most HCV genotypes has been challenging due to the antigenic diversity of the HCV genome. Here, we refined a high-yield subunit HCV vaccine that elicited broadly neutralizing antibody responses in preclinical trials. We found that soluble HCV E2 protein (sE2) produced in insect cells is distinctly glycosylated and is more immunogenic than sE2 produced in mammalian cells, suggesting that glycosylation patterns should be taken into consideration in efforts to generate antibody-based recombinant vaccines against HCV. We further showed that sE2 vaccination confers protection against HCV infection in a genetically humanized mouse model. Thus, our work identified a promising broadly protective HCV vaccine candidate that should be considered for further preclinical and clinical development.


Heterogeneous expression and release of CD14 by human gingival fibroblasts: characterization and CD14-mediated interleukin-8 secretion in response to lipopolysaccharide.
To identify the role in periodontal inflammatory diseases of human gingival fibroblasts (HGF), the major constituents of gingival tissue, the expression of CD14, a possible lipopolysaccharide (LPS) receptor, and the release of soluble CD14 (sCD14) by HGF were examined. Among the HGF samples from the nine donors tested, more than 50% of the HGF from five donors expressed CD14 but less than 20% of HGF from the other four donors did so, as determined by flow cytometric analysis. The CD14 expression on the cell surface was correlated with the expression of CD14 mRNA. The HGF and skin and lung fibroblasts tested expressed no CD18, which indicates that fibroblasts do not possess other LPS receptors, such as CD11b/CD18 and CD11c/CD18. The CD14 expression by the HGF was decreased after subculturing and was highest at the confluent stage of culture. The treatment of high-CD14-expressing (CD14(high)) HGF with phosphatidylinositol-phospholipase C reduced CD14 expression; this result and the increase in a 55-kDa CD14 indicate that the membrane CD14 (mCD14) on the HGF may be a 55-kDa glycosylphosphatidylinositol-anchored protein. CD14(high) HGF spontaneously released 48- and 57-kDa sCD14. The total release of sCD14 by the HGF was augmented by gamma interferon and Escherichia coli LPS in accordance with the increased expression of mCD14. The CD14(high) HGF secreted interleukin-8 in response to LPS, and the secretion was completely inhibited by anti-CD14 antibody. These results suggest that (i) HGF consist of populations that are heterogeneous on the basis of different levels of expression of CD14 and (ii) CD14(high) HGF secrete inflammatory cytokines in response to LPS via CD14.


Effect of UV light disinfection on antibiotic-resistant coliforms in wastewater effluents.
Total coliforms and total coliforms resistant to streptomycin, tetracycline, or chloramphenicol were isolated from filtered activated sludge effluents before and after UV light irradiation. Although the UV irradiation effectively disinfected the wastewater effluent, the percentage of the total surviving coliform population resistant to tetracycline or chloramphenicol was significantly higher than the percentage of the total coliform population resistant to those antibiotics before UV irradiation. This finding was attributed to the mechanism of R-factor-mediated resistance to tetracycline. No significant difference was noted for the percentage of the surviving total coliform population resistant to streptomycin before or after UV irradiation. Multiple drug resistance patterns of 300 total coliform isolates revealed that 82% were resistant to two or more antibiotics. Furthermore, 46% of these isolates were capable of transferring antibiotic resistance to a sensitive strain of Escherichia coli.


Purely nonlinear disorder-induced localizations and their parametric amplification.
We investigate spatial localization in a quadratic nonlinear medium in the presence of randomness. By means of numerical simulations and theoretical analyses we show that, in the down conversion regime, the transverse random modulation of the nonlinear susceptibility generates localizations of the fundamental wave that grow exponentially in propagation. The localization length is optically controlled by the pump intensity that determines the amplification rate. The results also apply to cubic nonlinearities.


Comparison of spherical and realistically shaped boundary element head models for transcranial magnetic stimulation navigation.
MRI-guided real-time transcranial magnetic stimulation (TMS) navigators that apply electromagnetic modeling have improved the utility of TMS. However, their accuracy and speed depends on the assumed volume conductor geometry. Spherical models found in present navigators are computationally fast but may be inaccurate in some areas. Realistically shaped boundary-element models (BEMs) could increase accuracy at a moderate computational cost, but it is unknown which model features have the largest influence on accuracy. Thus, we compared different types of spherical models and BEMs. Globally and locally fitted spherical models and different BEMs with either one or three compartments and with different skull-to-brain conductivity ratios (1/1-1/80) were compared against a reference BEM. The one-compartment BEM at inner skull surface was almost as accurate as the reference BEM. Skull/brain conductivity ratio in the range 1/10-1/80 had only a minor influence. BEMs were superior to spherical models especially in frontal and temporal areas (up to 20mm localization and 40% intensity improvement); in motor cortex all models provided similar results. One-compartment BEMs offer a good balance between accuracy and computational cost. Realistically shaped BEMs may increase TMS navigation accuracy in several brain areas, such as in prefrontal regions often targeted in clinical applications.


Effects of glycine supplementation at varying levels of methionine and cystine on the growth performance of broilers fed reduced crude protein diets.
Two experiments were conducted to investigate Gly addition to reduced crude protein corn-soybean meal (C-SBM) diets with varying levels of TSAA achieved by varying Met and Cys. The experiments were conducted with female Ross 708 broilers in brooder batteries from 0 to 18 d posthatching. Treatments had 6 replicates with 6 broilers/pen. Diets in all experiments were fed without or with Gly supplementation to contain 2.32% total Gly + Ser. All diets were C-SBM based and formulated to contained 1.27% standardized ileal digestible Lys supplemented with 0.20% Lys (0.394% Lys·SO(4)) and to meet or exceed the requirement of all nutrients except Met and Cys where appropriate. Experiment 1 consisted of 8 dietary treatments. Three ratios of Met to Cys (60:40, 50:50, and 40:60) were used on a mole for mole basis to achieve 0.063 mol of TSAA/kg of feed and a positive control with Met:Cys of 50:50 at 0.76 TSAA:Lys. Glycine supplementation did not affect ADG or ADFI; however, G:F was increased (P = 0.003) with Gly supplementation. An increase in Cys and a decrease in Met resulted in a decrease (P = 0.028) in ADG but had no effect on ADFI or G:F. In experiment 2, Met was kept constant at a marginal level of 0.45% and Cys was increased in 0.05% increments from 0.35 to 0.50%. Glycine supplementation had no main effect on ADG, ADFI, or G:F; however, Gly increased G:F at the lower levels of Cys but not at the higher levels (Gly × Cys, P = 0.031). A linear decrease (P = 0.071) was found in ADFI with increasing Cys supplementation. These data indicate that Gly increased G:F in female broilers fed suboptimal levels of Met and Cys but not at Cys levels at or above the requirement. This implies that the synthesis of Cys accounts for a portion of the increased G:F observed from Gly supplementation in female broilers fed reduced CP C-SBM diets.


Port Access (Thru-Port System) video-assisted mitral valve surgery.
The aim of this review is to focus the attention on the "state of the art" of port-access and video-assisted mitral valve surgery appraising the results compared to the traditional approach, describing different technical strategies and analyzing how to avoid and manage its related complications concomitantly elucidating which procedure is associated with the most favorable risk-benefit and cost-benefit profile. Our default strategy is to use the minimally invasive approach combining video-assisted right mini-thoracotomy with endo-cardiopulmonary bypass and endo-aortic balloon occlusion (EBO) whenever possible. This choice is supported by the evidence that, after an initial learning curve, it is a safe and effective approach in terms of short- and long-term results, mainly for redo operations and even for elderly patients with moderately elevated peri-operative risk.


Mechanisms of placebo analgesia: A dual-process model informed by insights from cross-species comparisons.
Placebo treatments are pharmacologically inert, but are known to alleviate symptoms across a variety of clinical conditions. Associative learning and cognitive expectations both play important roles in placebo responses, however we are just beginning to understand how interactions between these processes lead to powerful effects. Here, we review the psychological principles underlying placebo effects and our current understanding of their brain bases, focusing on studies demonstrating both the importance of cognitive expectations and those that demonstrate expectancy-independent associative learning. To account for both forms of placebo analgesia, we propose a dual-process model in which flexible, contextually driven cognitive schemas and attributions guide associative learning processes that produce stable, long-term placebo effects. According to this model, the placebo-induction paradigms with the most powerful effects are those that combine reinforcement (e.g., the experience of reduced pain after placebo treatment) with suggestions and context cues that disambiguate learning by attributing perceived benefit to the placebo. Using this model as a conceptual scaffold, we review and compare neurobiological systems identified in both human studies of placebo analgesia and behavioral pain modulation in rodents. We identify substantial overlap between the circuits involved in human placebo analgesia and those that mediate multiple forms of context-based modulation of pain behavior in rodents, including forebrain-brainstem pathways and opioid and cannabinoid systems in particular. This overlap suggests that placebo effects are part of a set of adaptive mechanisms for shaping nociceptive signaling based on its information value and anticipated optimal response in a given behavioral context.


Effects of acute myocardial ischemia on intramyocardial contraction heterogeneity: A study performed with ultrasound integrated backscatter during transesophageal atrial pacing.
[corrected] Subendocardial thickening is greater than subepicardial thickening and acute myocardial ischemia mainly impairs the former. Integrated backscatter cyclic variations (IBScv) reflect regional myocardial contractility and are blunted during myocardial ischemia. We hypothesized that stress-induced myocardial ischemia mainly affects subendocardial IBScv. Multiplane transesophageal echocardiography and simultaneous atrial pacing were performed in 12 patients without coronary artery disease (CAD) and in 25 with significant CAD. In a transgastric 2-chamber view, we calculated IBScv in subendocardium and subepicardium and a heterogeneity index, both at rest and at peak-pacing. In 27 myocardial segments of patients with normal coronary arteries, and in 16 myocardial segments supplied by coronary artery without significant stenosis in patients with CAD, there was a transmural gradient of IBScv at rest and the heterogeneity index did not change during all the protocol steps. In the 53 myocardial segments related to a significantly narrowed coronary artery, the transmural gradient of IBScv, present at rest, significantly decreased at peak-pacing because of subendocardial blunting, but promptly recovered 5 seconds after pacing interruption. Moreover, the myocardial thickening at rest and peak pacing correlated with the subendocardial IBScv behavior and not with the subepicardial one. IBScv are greater in the subendocardium than in the subepicardium. Atrial pacing stress test does not affect IBScv in segments supplied by nonstenotic coronary arteries, whereas it affects segments supplied by diseased coronary arteries, blunting exclusively subendocardial IBScv. Heterogeneity of IBScv intramyocardial changes caused by stress-induced ischemia must be taken into account when using IBScv for investigating myocardial ischemia.


Role of tumor necrosis factor-alpha and matrix metalloproteinase-9 in blood-brain barrier disruption after peripheral thermal injury in rats.
A relationship has been found between peripheral thermal injury and cerebral complications leading to injury and death. In the present study, the authors examined whether tumor necrosis factor-alpha (TNF-alpha) and matrix metalloproteinase-9 (MMP-9) play a causative role in blood-brain barrier (BBB) disruption after peripheral thermal injury. Thirty-two male Sprague-Dawley rats were subjected to thermal injury. One hour later, 8 rats were injected with TNF-alpha neutralizing antibody, and 8 were injected with doxycycline, an inhibitor of the MMP family proteins; 16 rats did not receive any treatment. Brain tissue samples obtained 7 hours after injury in the treated animals were examined for BBB function by using fluorescein isothiocyanate-dextran and by assessing parenchymal water content. Protein expression of basement membrane components (collagen IV, laminin, and fibronectin) was quantified on Western blot analysis, and MMP-9 protein expression and enzyme activity were determined using Western blot and gelatin zymography. Thermally injured rats that did not receive treatment were killed at 3, 7, or 24 hours after injury and tested for BBB functioning at each time point. Histological analysis for basement membrane proteins was also conducted in untreated rats killed at 7 hours after injury. Results of testing in injured rats were compared with those obtained in a control group of rats that did not undergo thermal injury. At 7 hours after thermal injury, a significant increase in the fluorescein isothiocyanate-dextran and water content of the brain was found (p < 0.05), but BBB dysfunction was significantly decreased in the rats that received TNF-alpha antibody or doxycycline (p < 0.05). In addition, the components of the basal lamina were significantly decreased at 7 hours after thermal injury (p < 0.01), and there were significant increases in MMP-9 protein expression and enzyme activity (p < 0.05). The basal lamina damage was reversed by inhibition of TNF-alpha and MMP-9, and the increase in MMP-9 protein was reduced in the presence of doxycycline (p < 0.05). The authors found that MMP-9 enzyme activity was significantly increased after thermal injury (p < 0.01) but decreased in the presence of either TNF-alpha antibody or doxycycline (p < 0.01). The dual, inhibitory activity of both TNF-alpha and MMP-9 in brain injury suggests that a TNF-alpha and MMP-9 cascade may play a key role in BBB disruption. These results offer a better understanding of the pathophysiology of burn injuries, which may open new avenues for burn treatment beyond the level of current therapies.


Medicare: where to now?
Although it is now more than ten years since Medicare was introduced, Medicare is still a relevant response to ensuring access to health care. However, marginal improvements to Medicare are needed in three areas. First, in terms of bureaucratic issues, rational development of health policy suffers from problems associated with the contemporary division of Commonwealth and State responsibilities. Secondly, more sophisticated methods for priority-setting, which take account of both the technical and allocative efficiency issues, are needed. Finally, Medicare needs to be enhanced by improving its ability to meet the diverse and varying needs of patients with long-term illnesses: improvement in 'coordinated care' is necessary. Approaches to meeting these needs are canvassed.


Integrated externally and internally generated task predictions jointly guide cognitive control in prefrontal cortex.
Cognitive control proactively configures information processing to suit expected task demands. Predictions of forthcoming demand can be driven by explicit external cues or be generated internally, based on past experience (cognitive history). However, it is not known whether and how the brain reconciles these two sources of information to guide control. Pairing a probabilistic task-switching paradigm with computational modeling, we found that external and internally generated predictions jointly guide task preparation, with a bias for internal predictions. Using model-based neuroimaging, we then show that the two sources of task prediction are integrated in dorsolateral prefrontal cortex, and jointly inform a representation of the likelihood of a change in task demand, encoded in frontoparietal cortex. Upon task-stimulus onset, dorsomedial prefrontal cortex encoded the need for reactive task-set adjustment. These data reveal how the human brain integrates external cues and cognitive history to prepare for an upcoming task.


Oral immunization with recombinant Streptococcus gordonii expressing porphyromonas gingivalis FimA domains.
Porphyromonas gingivalis, a gram-negative anaerobe, is implicated in the etiology of adult periodontitis. P. gingivalis fimbriae are one of several critical surface virulence factors involved in both bacterial adherence and inflammation. P. gingivalis fimbrillin (FimA), the major subunit protein of fimbriae, is considered an important antigen for vaccine development against P. gingivalis-associated periodontitis. We have previously shown that biologically active domains of P. gingivalis fimbrillin can be expressed on the surface of the human commensal bacterium Streptococcus gordonii. In this study, we examined the effects of oral coimmunization of germfree rats with two S. gordonii recombinants expressing N (residues 55 to 145)- and C (residues 226 to 337)-terminal epitopes of P. gingivalis FimA to elicit FimA-specific immune responses. The effectiveness of immunization in protecting against alveolar bone loss following P. gingivalis infection was also evaluated. The results of this study show that the oral delivery of P. gingivalis FimA epitopes via S. gordonii vectors resulted in the induction of FimA-specific serum (immunoglobulin G [IgG] and IgA) and salivary (IgA) antibody responses and that the immune responses were protective against subsequent P. gingivalis-induced alveolar bone loss. These results support the potential usefulness of the S. gordonii vectors expressing P. gingivalis fimbrillin as a mucosal vaccine against adult periodontitis.


Symmetry of initial cell divisions among primitive hematopoietic progenitors is independent of ontogenic age and regulatory molecules.
We have developed a time-lapse camera system to follow the replication history and the fate of hematopoietic stem cells (HSC) at a single-cell level. Combined with single-cell culture, we correlated the early replication behavior with colony development after 14 days. The membrane dye PKH26 was used to monitor cell division. In addition to multiple, synchronous, and symmetric divisions, single-sorted CD34(+)/CD38(-) cells derived from fetal liver (FLV) also gave rise to a daughter cell that remained quiescent for up to 8 days, whereas the other daughter cell proliferated exponentially. Upon separation and replating as single cells onto medium containing a cytokine cocktail, 60.6% +/- 9.8% of the initially quiescent cells (PKH26 bright) gave rise again to colonies and 15.8% +/- 7.8% to blast colonies that could be replated. We have then determined the effects of various regulatory molecules on symmetry of initial cell divisions. After single-cell sorting, the CD34(+)/CD38(-) cells derived from FLV were exposed to flt3-ligand, thrombopoietin, stem cell factor (SCF), or medium containing a cytokine cocktail (with SCF, interleukin-3, interleukin-6, granulocyte-macrophage colony-stimulating factor, and erythropoietin). Whereas mitotic rate, colony efficiency, and asymmetric divisions could be altered using various regulatory molecules, the asymmetric division index, defined as the number of asymmetric divisions versus the number of dividing cells, was not altered significantly. This observation suggests that, although lineage commitment and cell proliferation can be skewed by extrinsic signaling, symmetry of early divisions is probably under the control of intrinsic factors.


Mineralocorticoid-specificity of aldosterone-induced protein synthesis in giant-toad (Bufo marinus) urinary bladders.
We have identified a group of proteins (Mr approximately 70000-80000; pI approximately 5.8-6.4) in giant-toad (Bufo marinus) urinary-bladder epithelial cells whose synthesis appears to be related to aldosterone-stimulated Na+ transport. To define this relationship further, we examined whether submaximal natriferic concentrations of aldosterone induced these proteins and whether spironolactone (a specific mineralocorticoid antagonist in renal epithelia) inhibited their synthesis. Short-circuit current was used to measure Na+ transport and epithelial-cell protein synthesis was detected with high-resolution two-dimensional polyacrylamide-gel electrophoresis and autoradiography. Submaximal natriferic concentrations of aldosterone (1.4 X 10(-8) M) induced the same proteins as maximal concentrations of the hormone (1.4 X 10(-7) M). In contrast, in previous experiments, similar proteins were not induced by subnatriferic concentrations (5.0 X 10(-8) M) of cortisol, a glucocorticoid. A spironolactone/aldosterone molar ratio of 2000:1 was required to inhibit aldosterone-stimulated Na+ transport completely; ratios of 200:1 and 500:1 produced partial inhibition. Concentrations of spironolactone that abolished aldosterone-stimulated Na+ transport also inhibited aldosterone-induced protein synthesis. We conclude that the synthesis of the proteins we have identified is specifically related to activation of the mineralocorticoid pathway.


The universe of galectin-binding partners and their functions in health and disease.
Galectins, a family of evolutionarily conserved glycan-binding proteins, play key roles in diverse biological processes including tissue repair, adipogenesis, immune cell homeostasis, angiogenesis, and pathogen recognition. Dysregulation of galectins and their ligands has been observed in a wide range of pathologic conditions including cancer, autoimmune inflammation, infection, fibrosis, and metabolic disorders. Through protein-glycan or protein-protein interactions, these endogenous lectins can shape the initiation, perpetuation, and resolution of these processes, suggesting their potential roles in disease monitoring and treatment. However, despite considerable progress, a full understanding of the biology and therapeutic potential of galectins has not been reached due to their diversity, multiplicity of cell targets, and receptor promiscuity. In this article, we discuss the multiple galectin-binding partners present in different cell types, focusing on their contributions to selected physiologic and pathologic settings. Understanding the molecular bases of galectin-ligand interactions, particularly their glycan-dependency, the biochemical nature of selected receptors, and underlying signaling events, might contribute to designing rational therapeutic strategies to control a broad range of pathologic conditions.


Trade routes and plague transmission in pre-industrial Europe.
Numerous historical works have mentioned that trade routes were to blame for the spread of plague in European history, yet this relationship has never been tested by quantitative evidence. Here, we resolve the hypothetical role of trade routes through statistical analysis on the geo-referenced major trade routes in the early modern period and the 6,656 geo-referenced plague outbreak records in AD1347-1760. Ordinary Least Square (OLS) estimation results show that major trade routes played a dominant role in spreading plague in pre-industrial Europe. Furthermore, the negative correlation between plague outbreaks and their distance from major trade ports indicates the absence of a permanent plague focus in the inland areas of Europe. Major trade routes decided the major plague outbreak hotspots, while navigable rivers determined the geographic pattern of sporadic plague cases. A case study in Germany indicates that plague penetrated further into Europe through the local trade route network. Based on our findings, we propose the mechanism of plague transmission in historical Europe, which is imperative in demonstrating how pandemics were spread in recent human history.


Optical associatron: a simple model for optical associative memory.
A new neural network architecture for optical computing is proposed. This architecture can be used for implementing a simple system based on a modified theory of the associative memories. It is shown that the memorized patterns can be recalled perfectly by an experimental system using two microchannel spatial light modulators. The modified theory and principle of operation of the optical associatron system are discussed, and the system itself and basic experimental results are also described.


Continuing education in informatics among registered nurses in the United States in 2000.
Continuing education is one method for acquiring competency in informatics among nurses. However, little is known about nurses' participation in continuing education in informatics and the factors or characteristics that motivate them to pursue this type of education. This article identifies the proportion and characteristics of U.S. registered nurses reporting continuing education in informatics in 2000. A secondary data analysis was conducted with data from the National Sample Survey of Registered Nurses. More than 25,000 nurses responded to this survey in 2000. Bivariate and logistic regression analyses were conducted to examine the association of reported continuing education in informatics with demographic, educational, and other characteristics of respondents to the survey. Of the respondents, 21% reported continuing education in informatics in the year before the survey. The probability of continuing education in informatics increased with Internet access and decreased for nurses working outside hospitals or providing direct patient care. Relatively low proportions of registered nurses report continuing education in informatics, but some opportunities exist to increase rates.


Malignant hyperthermia testing in probands without adverse anesthetic reaction.
Malignant hyperthermia (MH) is triggered by reactions to anesthetics. Reports link nonanesthetic-induced MH-like reactions to a variety of disorders. The objective of the authors was to retrospectively investigate the reasons for referrals for MH testing in nonanesthetic cases and assess their phenotype. In addition, the response to the administration of oral dantrolene in nonanesthetic probands with positive caffeine-halothane contracture test (CHCT) was investigated. Following institutional research ethics board approval, probands without reaction to anesthesia, who underwent CHCT, were selected. Clinical details and response to dantrolene were analyzed. In total, 87 of 136 (64%) patients referred for nonanesthetic indications tested positive to the CHCT. Of these, 47 with a high creatine kinase (CK), 9 with exercise-induced rhabdomyolysis and/or exercise intolerance, 2 with high CK and exercise-induced rhabdomyolysis and/or exercise intolerance, 15 with postviral chronic fatigue, and 14 with muscle weakness of unknown etiology had a positive CHCT. These patients had a higher CK compared with those with negative CHCT. Oral dantrolene improved the musculoskeletal symptoms in 28 of 34 (82%) CHCT-positive patients. Response to treatment was associated with a significantly higher pretreatment CK and a greater posttreatment CK reduction. A positive CHCT may represent more than simply an anesthetic-related disorder. Individuals with positive CHCTs may exhibit muscle symptoms without exposure to MH-triggering anesthetics. Oral dantrolene may be useful in alleviating these symptoms.


Preexposure prophylaxis for adolescents and young adults at risk for HIV infection: is an ounce of prevention worth a pound of cure?
An alarming proportion of incident human immunodeficiency virus (HIV) infections worldwide occur in youth. In the United States, 69% of all new infections among youth occurred in young men who have sex with men (YMSM). Recent studies show the promise of preexposure prophylaxis (PrEP) for preventing HIV infection, but research efforts suffer from disproportionately low representation of the youth who are most at risk. Youth-focused research is critical and should include behavioral, community, and biomedical interventions to create a comprehensive HIV prevention package. The many ethical, legal, and regulatory considerations in conducting HIV research among, and in providing care services to, youth must be addressed so that those at high risk and most likely to benefit can have unfettered access to safe and effective health-promoting interventions. YMSM and minority youth are at substantial HIV risk and urgently need effective HIV prevention tools for which the short and long-term benefits and risks have been carefully considered.


Use of suboptimal control arms in randomized clinical trials of investigational cancer drugs in China, 2016-2021: An observational study.
The use of suboptimal controls in randomized trials of new cancer drugs can produce potentially unreliable clinical efficacy results over the current standard of care and expose patients to substandard therapy. We aim to investigate the proportion of randomized trials of investigational cancer drugs that used a suboptimal control arm and the number of trial participants at risk of exposure to suboptimal treatments in China. The association between the use of a suboptimal control and concluding statistical significance on the primary endpoint was also examined. This observational study included randomized controlled trials (RCTs) of cancer drugs that were authorized by specific Chinese institutional review boards between 2016 and 2021, supporting investigational new drug applications of these drugs in China. The proportion of trials that used a suboptimal control arm and the total number of trial participants at risk of exposure to suboptimal treatments were calculated. In a randomized trial for a specific condition, a comparator was deemed suboptimal if it was not recommended by clinical guidelines published in priori or if there existed a regimen with a higher level of recommendation for the indication. The final sample included 453 Phase II/III and Phase III randomized oncology trials. Overall, 60 trials (13.2%) adopted a suboptimal control arm. Among them, 58.3% (35/60) used comparators that were not recommended by a prior guideline for the indication. The cumulative number of trial participants at risk of exposure to suboptimal treatments totaled 18,610 by the end of 2021, contributing 15.1% to the total number of enrollees of all sampled RCTs in this study. After adjusting for the year of ethical approval, region of participant recruitment, line of therapy, and cancer site, second-line therapies (adjusted odds ratio [aOR] = 2.7, 95%CI [1.2, 5.9]), adjuvant therapies (aOR = 8.9, 95% CI [3.4, 23.1]), maintenance therapies (aOR = 5.2, 95% CI [1.6, 17.0]), and trials recruiting participants in China only (aOR = 4.1, 95% CI [2.1, 8.0]) were more likely to adopt a suboptimal control. For the 105 trials with publicly available results, no statistically significant difference was observed between the use of a suboptimal control and concluding positive on the primary endpoint (100.0% [12/12] versus 83.9% [78/93], p = 0.208). The main limitation of this study is its reliance on clinical guidelines that could vary across cancer types and time in assessing the quality of the control groups. In this study, over one-eighth of randomized trials of cancer drugs registered to apply for regulatory approval in China used a suboptimal comparator. Our results highlight the necessity to refine the design of randomized trials to generate optimal clinical evidence for new cancer therapies.


[Adenovirus-mediated transfer of TIMP-4 gene inhibits neointimal formation after balloon injury].
To investigate the effects of human tissue inhibitor of matelloproteinase-4 (TIMP-4) on vascular smooth muscle cells (VSMCs) migration and the neointimal formation after balloon injury in rats. The cultured VSMCs were transfected with an adenoviral vector containing human TIMP-4 gene, AdTIMP-4. Effect of TIMP-4 on VSMC migration was investigated by monolayer cell scrape. AdTIMP-4, control adenoviral vector or PBS was transduced into the rat carotid artery from the adventitial after carotid artery injury. Cell number within the internal elastic lamina 4 days after gene transfer was counted and neointima/media area ratio 28 days after gene transfer was calculated. The migration distance of VSMCs infected with AdTIMP-4 was inhibited markedly. Morphometric analysis demonstrated a statistically significant reduction in the number of cells migrated into neointima compared with controls [(32.5 +/- 4.8) cells per section, (33.8 +/- 7.0) cells per section and (8.2 +/- 2.4) cells per section for uninfected, AdGFP-treated and AdTIMP-4-treated arteries, respectively]. There was a reduction of intima/media ratio of TIMP-4-treated group by 66.5% compared with control groups 28 days after gene transfer (P < 0.01). There was no statistical difference between the control adenoviral vector group and PBS group. Adenoviral mediated gene transfer of TIMP-4 inhibits migration of VSMC and significantly reduces neointimal hyperplasia in the rat carotid balloon injury model. The TIMP-4 gene transfer is a potential therapeutic approach to preventing neointimal formation after balloon injury.


Important role of melanin for fertility in the fungus Podospora anserina.
Melanins are pigments used by fungi to withstand various stresses and to strengthen vegetative and reproductive structures. In Sordariales fungi, their biosynthesis starts with a condensation step catalyzed by an evolutionary-conserved polyketide synthase. Here we show that complete inactivation of this enzyme in the model ascomycete Podospora anserina through targeted deletion of the PaPks1 gene results in reduced female fertility, in contrast to a previously analyzed nonsense mutation in the same gene that retains full fertility. We also show the utility of PaPks1 mutants for detecting rare genetic events in P. anserina, such as parasexuality and possible fertilization and/or apomixis of nuclei devoid of mating-type gene.


More than skin-deep: the many dimensions of the psoriatic disease.
Psoriasis is among the most common skin diseases, exhibiting a wide spectrum of clinical manifestations. Joint involvement in the form of psoriatic arthritis is readily recognised, but both frequency and severity of this manifestation have long been underestimated. More recently, additional important diseases have been found to be associated with psoriasis, including the metabolic syndrome (or components thereof), cardiovascular diseases, lymphoma, and anxiety/depression. In the past, psoriasis treatment aimed at suppressing acute rashes. Current concepts regard psoriasis as a chronic systemic inflammatory condition and cardiovascular risk factor. In the light of this concept, long-term disease control through systemic maintenance therapy is increasingly recommended by experts. This approach became feasible with the approval of numerous biologics for the treatment of psoriasis. But to really address all medical needs of psoriasis patients, a truly interdisciplinary, comprehensive management approach is needed. Several national societies have already published algorithms to ensure that this need will be implemented in the daily practice of dermatologists and nondermatologists alike.


Biosynthesis of Plant Triterpenoid Saponins in Microbial Cell Factories.
Triterpenoid saponins are triterpenoid glycoside compounds which have been widely used in pharmaceutical, agricultural, and food industries. Traditionally, they are extracted from plants, which is time-consuming and environmentally unfriendly. Recently, de novo synthesis of triterpenoid saponins in microbial cell factories was realized, which provides a promising and green approach to alter the traditional supply way. However, the complex biosynthetic pathway and the poor suitability between the endogenous and heterogeneous pathways tremendously limit the yield of triterpenoid saponins. We introduce the biosynthetic pathways of triterpenoid saponins first, and we then summarize the microbial cell factories developed to produce these compounds. Further, we discuss the strategies applied to enhance the production. This paper systematically illustrates the biosynthesis of plant triterpenoid saponins in microbial cell factories.


Retrotransposon insertion polymorphisms in six rice genes and their evolutionary history.
Retrotransposons are abundant in higher plant genomes. Although retrotransposons associated with plant genes have been identified, little is known about their evolutionary conservation at the level of species and subspecies. In the present study, we investigated the phylogenetic distribution of long terminal repeat (LTR) retrotransposon, long interspersed nuclear element (LINE) and short interspersed nuclear element (SINE) insertions in six genes in 95 cultivated and wild rice genotypes. These six genes are likely to be functional based on nonsynonymous (Ka) to synonymous (Ks) substitution ratios which were found to be significantly <1. Different conservation patterns of these retrotransposons in genes were observed in cultivated and wild rice species. Four out of seven retrotransposon insertions appear to predate the ancestral Oryza AA genome. Two of these insertions in genes 4 and 5 occurred early in the evolutionary history of Oryza. Two retrotransposon insertions in gene 1 arose after the divergence of Asian cultivated rice from its wild ancestor. Furthermore, the retrotransposon insertion in gene 3 appears to have occurred in the ancestral lineage leading to temperate japonicas. Conservation of retrotransposon insertions in genes in specific groups, species, and lineages might be related to their specific function.


Action-effect binding is decreased in motor conversion disorder: implications for sense of agency.
The abnormal movements seen in motor conversion disorder are affected by distraction and entrainment, similar to voluntary movement. Unlike voluntary movement, however, patients lack a sense of control for the abnormal movements, a failure of "self-agency." The action-effect binding paradigm has been used to quantify the sense of self-agency, because subjective contraction of time between an action and its effect only occurs if the patient feels that they are the agent responsible for the action. We used this paradigm, coupled with emotional stimuli, to investigate the sense of agency with voluntary movements in patients with motor conversion disorder. Twenty patients with motor conversion disorder and 20 age-matched and sex-matched healthy volunteers used a rotating clock to judge the time of their own voluntary key presses (action) and a subsequent auditory tone (effect) after they completed conditioning blocks in which high, medium, and low tones were coupled to images of happy, fearful, and neutral faces. The results replicated those produced previously: it was reported that an effect after a voluntary action occurred earlier, and the preceding action occurred later, compared with trials that used only key presses or tones. Patients had reduced overall binding scores relative to healthy volunteers, suggesting a reduced sense of agency. There was no effect of the emotional stimuli (faces) or other interaction effects. Healthy volunteers with subclinical depressive symptoms had higher overall binding scores. We demonstrate that patients with motor conversion disorder have decreased action-effect binding for normal voluntary movements compared with healthy volunteers, consistent with the greater experience of lack of control.


Sexual concurrency among young African American women.
Young African-American women are disproportionately affected by human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) sexually transmitted infections (STI), and engage in greater sexual concurrency than other race/ethnicities. It is important to evaluate behaviors and characteristics associated with the risk of sexual concurrency, so that interventions can target factors most likely to affect positive change. An emphasis on correlates of concurrency beyond individual-level factors has been suggested. The purpose of this study, therefore, was to identify individual- and partner-level characteristics associated with sexual concurrency among high-risk, young African-American women. Data were collected from 570 African-American adolescent women (aged 15-21) recruited from a STI clinic, a family planning clinic, and a teen clinic located in Atlanta, GA from March 2002 through August 2004. Logistic regression analysis was conducted in 2012 to evaluate correlates of sexual concurrency. Results show that almost one-quarter of participants reported sexually concurrent partnerships and 28.4% suspected male partner concurrency. Logistic regression results indicated the number of lifetime sexual partners and relationship factors were the primary contributors to engaging in concurrency in this sample. These findings suggest relationship factors may be important contributors to the prevalence of sexual concurrency among young African-American women. Interventions targeted toward sexual health among young African-American women may need to specifically address partner/relationship factors. Through these findings, we hope to better understand sexual risk taking and develop strategies that would overcome barriers to existing interventions aimed at improving the sexual health outcomes of young African-American women.


Silicon photonic grating-assisted, contra-directional couplers.
We demonstrate, in both theory and experiment, 4-port, electrically tunable photonic filters using silicon contra-directional couplers (contra-DCs) with uniform and phase-shifted waveguide Bragg gratings. Numerical analysis, including both intra- and inter-waveguide couplings, is performed using coupled-mode theory and the transfer-matrix method. The contra-DC devices were fabricated by a CMOS-photonics manufacturing foundry and are electrically tunable using free-carrier injection. A 4-port, grating-based photonic resonator has been obtained using the phase-shifted contra-DC, showing a resonant peak with a 3-dB bandwidth of 0.2 nm and an extinction ratio of 24 dB. These contra-DC devices enable on-chip integration of Bragg-grating-defined functions without using circulators and have great potential for applications such as wavelength-division multiplexing networks and optical signal processing.


Effects of forest thinning on sap flow dynamics and transpiration in a Japanese cedar forest.
Historically, forest thinning in Japan was conducted to obtain high-quality timber from plantations. Today, in contrast, thinning is also motivated by forest water balance and climate change considerations. It is in this context that the present study examines the effects of thinning on the ecophysiological responses of remaining trees, which are inadequately understood, especially in relation to changes in the magnitude and duration of transpiration. Sap flux densities were measured in both outer and inner sapwood to obtain stand-scale transpiration for two years in the pre-thinning state and three years in post-thinning. The effects of thinning on transpiration were quantitatively evaluated based on canopy conductance models. The larger increases in outer sap flux density were found in the first year after the treatment, while those in inner sap flux density were detected in the second and third years. The remaining trees required a few of years to adjust to improved light conditions of the lower crown, resulting in a delayed response of inner sap flux density. As a result of this lag, transpiration was reduced to 71 % of the pre-thinning condition in the first year, but transpiration recovered to the pre-thinning levels in the second and third years due to compensating contributions from inner sap flow. In terms of more accurately chronicling the thinning effect, the distribution of sap flux density with respect to its radial pattern, is necessary. Such measurements are key to more comprehensively examining the ecophysiological response of forest plantations to thinning and, ultimately, its effect on the forest water balance.


Approaches for Curriculum and Assessment in Leader and Leadership Education and Development Programs in American Medical Schools.
This article was migrated. The article was marked as recommended. Problem: There is a growing call to add leader and leadership education to undergraduate medical education (UME). Yet, there currently are no established standards, competencies, curricula, or requirements for UME leader and leadership education and development (LEAD) programs. The Uniformed Services University of the Health Sciences (USU) F. Edward Hébert School of Medicine LEAD program hosts annual Summit and Working Group meetings to address issues and to share experiences about LEAD programs. Approach: Based on survey results following the 2017 USU LEAD Summit, working group participants reported that the meeting was valuable, should be repeated, and should address the specific topics of curriculum and assessment. Therefore, the 2018 Summit's goal was for participants to share experiences, ideas, and ways forward regarding leader and leadership curricula and assessment measures for UME. Themes from working groups were compiled and reported. Outcomes: Themes within LEAD curriculum include: (1) what to teach: relevant knowledge, skills, and abilities/attitudes (KSA) for specific topics; (2) when to teach: a life-cycle program woven through UME into graduate medical education and beyond; and (3) how to teach: near peers, development of mentors, and near-term, practical applications of skills. Themes within LEAD assessment include: (1) what to assess: alignment with program goals and curriculum within a positive culture of assessment and trust; (2) when to assess: occur at times that are consistent with the learning objectives and curriculum to provide information on incremental "growth" of students and the program; (3) how to assess: use formative and summative, qualitative and quantitative measures that are reliable and valid. Next steps: Based on feedback from working group participants at the 2018 Summit, the USU LEAD team will host a third Summit in April 2019 focusing on leader and leadership education and development across the healthcare work force life cycle.


Physiological responses to food deprivation in the house sparrow, a species not adapted to prolonged fasting.
Many wild birds fast during reproduction, molting, migration, or because of limited food availability. Species that are adapted to fasting sequentially oxidize endogenous fuels in three discrete phases. We hypothesized that species not adapted to long fasts have truncated, but otherwise similar, phases of fasting, sequential changes in fuel oxidization, and similar changes in blood metabolites to fasting-adapted species. We tested salient predictions in house sparrows (Passer domesticus biblicus), a subspecies that is unable to tolerate more than ~32 h of fasting. Our main hypothesis was that fasting sparrows sequentially oxidize substrates in the order carbohydrates, lipids, and protein. We dosed 24 house sparrows with [(13)C]glucose, palmitic acid, or glycine and measured (13)CO(2) in their breath while they fasted for 24 h. To ascertain whether blood metabolite levels reflect fasting-induced changes in metabolic fuels, we also measured glucose, triacylglycerides, and β-hydroxybutyrate in the birds' blood. The results of both breath (13)CO(2) and plasma metabolite analyses did not support our hypothesis; i.e., that sparrows have the same metabolic responses characteristic of fasting-adapted species, but on a shorter time scale. Contrary to our main prediction, we found that recently assimilated (13)C-tracers were oxidized continuously in different patterns with no definite peaks corresponding to the three phases of fasting and also that changes in plasma metabolite levels accurately tracked the changes found by breath analysis. Notably, the rate of recently assimilated [(13)C]glycine oxidization was significantly higher (P < 0.001) than that of the other metabolic tracers at all postdosing intervals. We conclude that the inability of house sparrows to fast for longer than 32 h is likely related to their inability to accrue large lipid stores, separately oxidize different fuels, and/or spare protein during fasting.


Patient-specific record linkage between emergency department and hospital admission data for a cohort of people who inject drugs: methodological considerations for frequent presenters.
People who inject drugs (PWID) have been identified as frequent users of emergency department (ED) and hospital inpatient services. The specific challenges of record linkage in cohorts with numerous administrative health records occurring in close proximity are not well understood. Here, we present a method for patient-specific record linkage of ED and hospital admission data for a cohort of PWID. Data from 688 PWID were linked to two state-wide administrative health databases identifying all ED visits and hospital admissions for the cohort between January 2008 and June 2013. We linked patient-specific ED and hospital admissions data, using administrative date-time timestamps and pre-specified linkage criteria, to identify hospital admissions stemming from ED presentations for a given individual. The ability of standalone databases to identify linked ED visits or hospital admissions was examined. There were 3459 ED visits and 1877 hospital admissions identified during the study period. Thirty-four percent of ED visits were linked to hospital admissions. Most links had hospital admission timestamps in-between or identical to their ED visit timestamps (n = 1035, 87%). Allowing 24-h between ED visits and hospital admissions captured more linked records, but increased manual inspection requirements. In linked records (n = 1190), the ED 'departure status' variable correctly reflected subsequent hospital admission in only 68% of cases. The hospital 'admission type' variable was non-specific in identifying if a preceding ED visit had occurred. Linking ED visits with subsequent hospital admissions in PWID requires access to date and time variables for accurate temporal sorting, especially for same-day presentations. Selecting time-windows to capture linked records requires discretion. Researchers risk under-ascertainment of hospital admissions if using ED data alone.


Stroke is an emergency.
Stroke is an emergency. Ischemic stroke is similar to myocardial infarction in that the pathogenesis is loss of blood supply to the tissue, which can result in irreversible damage if blood flow is not restored quickly. Public education is needed to emphasize the warning signs of stroke. Patients should seek medical help immediately, using emergency transport systems. Therapy geared toward minimizing the damage from an acute stroke should be started without delay in the emergency room. This includes measures to protect brain tissue, support perfusion pressure, and minimize cerebral edema. Strategies for improving recovery should also begin immediately. All major medical centers need stroke teams and stroke units. Stroke prevention should be given high priority as a public health strategy. Risk factor management should be part of general health care and should begin in childhood, with emphasis on nutrition, exercise, weight control, and avoidance of tobacco. Health screening and early treatment of hypertension and hypercholesterolemia has decreased the incidence of stroke and heart disease, but these efforts need to be expanded to reach all segments of the population. Basic research has opened the door to new therapies aimed at re-establishing blood flow and limiting tissue damage. Clinical trials have already led to changes in stroke prevention, including studies of carotid endarterectomy and ticlopidine and warfarin therapy (for patients with atrial fibrillation). Trials in progress are testing the usefulness of ancrod, neuroprotective agents, antioxidant agents, anti-inflammatory agents, low-molecular-weight heparin, thrombolytic drugs, and angioplasty. Any delay starting therapy after an acute stroke will result in progressive, irreversible loss of brain tissue. Clinicians should remember that for a stroke patient, time is brain tissue.


Stoichiometry of a half-turnover of band 3, the chloride transport protein of human erythrocytes.
The kinetics of human red blood cell Cl transport have been studied under nonequilibrium conditions to determine whether or not an outward Cl gradient can recruit the transport protein from an inward-facing to an outward-facing configuration. Three kinds of evidence are consistent with this outward recruitment. First, the initial net Cl efflux into a Cl-free phosphate medium is independent of the intracellular Cl concentration in the range 20-170 mM. Second, an outward Cl gradient strongly enhances the inhibitory potency of DNDS (4,4'-dinitro-2,2'-stilbene disulfonate), which suggests that DNDS binds primarily to outward-facing states. Finally, we have estimated the number of Cl ions transported during the putative outward recruitment. Resealed red cell ghosts containing only 70 muM 36Cl were resuspended at 0 degrees C in a Cl-free, HCO3-free Na2SO4 medium. In the first 10 s, or approximately 10(6) Cl ions per ghost, followed by a much slower further loss of Cl. The rapid loss of 10(6) Cl ions per ghost, which is abolished by pretreatment with DIDS (4,4'-diisothiocyano-2,2'-stilbene disulfonate), appears to represent the Cl that is transported during the first half-turnover of the transport cycle. These data are strong evidence that the influx and efflux events in the catalytic cycle for anion transport do not take place simultaneously, and that the stoichiometry of the transport cycle is close to one pair of anions exchanged per band 3 monomer.


Aryl-Modified Pentamethyl Cyanine Dyes at the C2' Position: A Tunable Platform for Activatable Photosensitizers.
Pentamethyl cyanine dyes are promising fluorophores for fluorescence sensing and imaging. However, advanced biomedical applications require enhanced control of their excited-state properties. Herein, a synthetic approach for attaching aryl substituents at the C2' position of the thio-pentamethine cyanine (TCy5) dye structure is reported for the first time. C2'-aryl substitution enables the regulation of both the twisted intramolecular charge transfer (TICT) and photoinduced electron transfer (PET) mechanisms to be regulated in the excited state. Modulation of these mechanisms allows the design of a nitroreductase-activatable TCy5 fluorophore for hypoxic tumor photodynamic therapy and fluorescence imaging. These C2'-aryl TCy5 dyes provide a tunable platform for engineering cyanine dyes tailored to sophisticated biological applications, such as photodynamic therapy.


Paget's disease of the breast: the experience of the European Institute of Oncology and review of the literature.
Paget's disease of the breast is an uncommon presentation of breast malignancy, accounting for 1-3% of all the breast tumors and presents in different histopathologic patterns: in association with an underlying invasive or non invasive carcinoma, or without any underlying neoplasia. In the literature, different methods are used for the treatment. Mastectomy with or without axillary dissection has been considered as the standard treatment procedure for many years. Several studies have already shown that breast conservation with radiation therapy is an oncologically safe option. Regarding the axillary approach, several studies have documented the presence of positive sentinel lymph node even in Paget's disease alone. The objective of this study was to retrospectively analyze outcome of patients affected by Paget's breast disease and to define our institutional experience. Between May 1996 and February 2003, 114 patients with confirmed Paget's disease of the breast were retrieved and underwent surgery at the European Institute of Oncology of Milan, Italy. The median age of the patients was 54 years at the time of the diagnosis. In our study, the histopathological examination of the operated specimen revealed one hundred seven patients with Paget's disease associated with an underlying invasive or non invasive carcinoma, and seven patients without underlying carcinoma. Patients underwent either conservative breast surgery or mastectomy, with or without sentinel lymph node biopsy and/or axillary surgery. Each patient was evaluated after surgery at a multidisciplinary meeting to selecting systemic therapy. Seven patients had "pure" Paget's disease of the breast and one hundred seven had the disease associated with an underlying carcinoma. As surgical techniques 71 mastectomies and 43 breast conserving surgeries have been performed. Complete axillary dissection was done in patients with clinically positive lymph node and/or sentinel lymph node biopsy positive. Sentinel lymph node biopsy was performed in nineteen patients with invasive component and five were positive and underwent axillary dissection. Eleven sentinel lymph node biopsies were done in patients with non invasive component and none of them was positive. Adjuvant systemic therapies were based on the final tumor, node and metastasis stage: thirty patients received adjuvant chemotherapy alone, fourteen received endocrine treatment alone, twenty-six patients were evaluated to receive both chemo and endocrine therapy. The median duration of follow up was 73 months and was updated in the last 6 months. Five patients developed local recurrence, one had regional recurrence, another two had loco-regional recurrences and fourteen had distant metastasis as a first event. Malignancy-related deaths were censored in the statistical analyses cancer for and due to another tumor in eleven patients. Additionally, deaths were not related to malignancy totally in thirteen patients. Screening examination and imaging techniques are fundamental. Breast conserving surgery combined with breast irradiation for patients with invasive and non invasive breast carcinoma has become the treatment of first choice. All surgical conservative approaches should include the complete nipple-areolar complex and margins of resected specimen free of tumor. Thanks to the evolution of the conservative approach, good cosmetic result can be obtained. To be informed about the axillary lymph node status and to avoid the patient to have a second surgical approach, sentinel lymph node biopsy should be performed.


Investigating the Hypothesis of Stress System Dysregulation as a Risk Factor for Central Serous Chorioretinopathy: A Literature Mini-Review.
Background: This mini-review addressed the question "what do we know about the association between the dysregulation of stress systems (HPA axis and SAM) and the onset and prognosis of CSC in adult populations?" Methods: The literature mini-review was conducted through electronic searches using the PubMed, Web of Science, and Scopus databases. All published human and animal studies with both observational and experimental designs from 1966 to October 2018 were included. Results: Our search identified 229 reports, of which 32 articles were ultimately identified to be reviewed in this paper. Among these key articles, twenty-three were related to exogenous and/or endogenous high glucocorticoids as risk factors for CSC, seven were related to Type-A behavior and chronic psychological distress as risk factors for idiopathic CSC, and two were related to stress-induced animal models of CSC. Nineteen out of twenty-three studies in the first group reported a consistent association between high circulating corticosteroids and the onset and prognosis of CSC. Six out of seven studies in the second group reported a consistent association between stress-induced allostatic (over)load and the appearance of more- or less-severe CSC disorders, assuming that elevated circulating steroids may constitute a kind of risk factor for the eye through dysregulation of the HPA axis. All the selected studies reported HPA axis dysregulation as a possible common factor to explain the association between high circulating corticosteroids and CSC. In contrast, the involvement of the SAM system is only indirectly taken into consideration through the PA and HR measures and/or plasma and 24-h urinary catecholamine levels. Therefore, information regarding the involvement of SAM system dysregulation in the onset and prognosis of CSC is lacking. This observation is particularly relevant in view of the fact that animal models of CSC in monkeys are primarily induced by adrenergic hypertonia and that the course of experimental CSC is not further exacerbated by the administration of corticosteroids.


HPLC analysis of insulin degradation products from isolated hepatocytes. Effects of inhibitors suggest intracellular and extracellular pathways.
Isolated rat hepatocytes were incubated with A14-[125I]monoiodotyrosyl insulin for 30 min, and labeled material was extracted from the cells and incubation media. The medium and the cell extract were chromatographed on a Sephadex G-50 column, and radioactivity eluting in the position of intact insulin was concentrated and analyzed on HPLC. The HPLC analysis of the cell extract showed two major products eluting from the column at 19 and 23 min, whereas medium extracts showed one prominent product eluting at 14 min. Inclusion of chloroquine in the incubation blocked the formation of cellular products at 19 and 23 min and caused the accumulation of a product eluting at 41 min while not affecting the media products. After sulfitolysis all cellular products contained an intact A-chain. Dansylcadaverine increased media products and altered the cell-extracted product pattern such that it had a major peak at 14 min, similar to media. These results suggest that two pathways for insulin degradation exist within hepatocytes. The extracellular process forms products that are essentially unchanged by chloroquine and dansylcadaverine. The intracellular process is altered by chloroquine and apparently inhibited by dansylcadaverine.


Analysis of HCV-immunoglobulin isotype complexes provide new insights into antibody response to HCV.
Hepatitis C virus (HCV) is known for its ability to establish persistent infection and cause chronic hepatitis in most infected individuals. The antibody response to HCV in HCV-circulating immune complexes (CIC) is unknown. In the present study, we have characterized distinct changes in patterns of HCV-immunoglobulin (Ig) constituents with disease category, viral mutation and clinical markers. The number of samples positive for single HCV-Ig, HCV-IgG and HCV-IgA, HCV-IgM and HCV-IgA, HCV-IgM and HCV-IgG, HCV-IgM, HCV-IgG and HCV-IgA in 47 samples tested were 8 (17%), 1 (2.1%), 9 (19.1%), 4 (8.5%) and 17 (36.2%), respectively. The occurrence of HCV-IgM and HCV-IgA in combination of two isotypes of HCV-Ig became predominant. These results show that defective IgG in HCV-CIC may contribute to long-term viremia. Further analysis indicated that the frequency of HCV RNA/IgA-CIC in the abnormal aspartic aminotransferase (AST) group was significantly higher than that of the normal AST group, and HCV RNA/IgA-CIC frequency in the abnormal alanine aminotransferase (ALT) group was slightly higher than that in the normal ALT group. IgA complexes may reflect the damage degree of liver function during the course of HCV infection. We also found that there were more mutations in supernatant than in other constituents from single-strand conformation polymorphism (SSCP) analysis. Our results suggest that Ig-complexed virions and free virions may have different biological consequences, with the latter being elusive to immunological elimination. The findings in this study may provide some new insights into antibody response to HCV.


Medial orbitofrontal cortex codes relative rather than absolute value of financial rewards in humans.
Functional imaging studies in recent years have confirmed the involvement of orbitofrontal cortex (OFC) in human reward processing and have suggested that OFC responses are context-dependent. A seminal electrophysiological experiment in primates taught animals to associate abstract visual stimuli with differently valuable food rewards. Subsequently, pairs of these learned abstract stimuli were presented and firing of OFC neurons to the medium-value stimulus was measured. OFC firing was shown to depend on the relative value context. In this study, we developed a human analogue of this paradigm and scanned subjects using functional magnetic resonance imaging. The analysis compared neuronal responses to two superficially identical events, which differed only in terms of the preceding context. Medial OFC response to the same perceptual stimulus was greater when the stimulus predicted the more valuable of two rewards than when it predicted the less valuable. Additional responses were observed in other components of reward circuitry, the amygdala and ventral striatum. The central finding is consistent with the primate results and suggests that OFC neurons code relative rather than absolute reward value. Amygdala and striatal involvement in coding reward value is also consistent with recent functional imaging data. By using a simpler and less confounded paradigm than many functional imaging studies, we are able to demonstrate that relative financial reward value per se is coded in distinct subregions of an extended reward and decision-making network.


Prevalence of renal artery disease and its prognostic significance in patients undergoing coronary bypass grafting.
Several studies demonstrated the prognostic importance of renal failure and peripheral artery disease in patients undergoing coronary artery bypass grafting (CABG), but data regarding the prognostic value of renal artery disease in this context are scarce. We aimed to study the prevalence and prognostic value of renal artery disease in patients undergoing CABG. We assessed by duplex ultrasound the renal arteries of 429 consecutive patients who underwent CABG, of whom 401 had satisfactory imaging quality to detect >60% renal artery stenosis (RAS) and/or an elevated resistive index (ERI>0.80). Of the 401 subjects included (age 68±10 years, 83% men), 40 (10%) had RAS and 35 (9%) had ERI. Nine patients (2.2%) had both conditions. Patients were followed up for 12.4±7.0 months. The primary outcome was composite, including 30-day death, stroke, and/or myocardial infarction. In a multivariate model adjusted for age, gender, cardiovascular (CV) risk factors, renal function, chronic obstructive pulmonary disease, the use of off-pump CABG, CV co-morbidities, and drugs, the presence of ERI was strongly associated with the occurrence of the composite outcome (odds ratio 4.3, 95% confidence interval 1.7 to 9.9, p=0.0006). Similarly, ERI, not RAS, was significantly associated with the 30-day acute kidney disease and the midterm mortality, as well as fatal and nonfatal CV events. In conclusion, regardless of renal function and other factors, the renal resistive index is a strong predictor of CV and renal events after CABG. Renal duplex ultrasound can identify a subgroup of patients at high risk of CABG.


Projection of Target Drug Particle Size in Oral Formulations Using the Refined Developability Classification System (rDCS).
Dissolution limitations to oral absorption can occur if the time required for dissolution is longer than the transit time across the small intestine and/or if dissolution is slower than the drug's permeation through the gut wall. These limitations most often occur for poorly soluble drugs. A standard method for overcoming dissolution issues is to reduce the particle size of the (solid) drug. Building on the refined Developability Classification System (rDCS), this work establishes a novel set of equations with which the appropriate degree of particle size reduction needed to mitigate dissolution limitations to absorption can be calculated. According to the type of data available, the appropriate equation(s) for each situation can be applied. Three case examples are used to illustrate implementation of the equations: voriconazole, lemborexant and istradefylline. Although for voriconazole (rDCS Class I) target radius (rtarget) estimates indicate that particle size reduction is unnecessary, for lemborexant (rDCS Class I) a radius of ≤20 µm would be required to improve absorption. For istradefylline (rDCS Class IIb) the rtarget was approximately 12 µm. Results are commensurate with literature information for these three drugs, signaling that the equations are suitable for application to a wide variety of drug substances.


Improvement in Peer Assessment Rating scores after nonextraction, premolar extraction, and mandibular incisor extraction treatments in patients with Class I malocclusion.
Different treatment protocols implemented for correction of Class I malocclusion aim at achieving ideal occlusal characteristics. This study was planned to evaluate the improvement in the occlusal characteristics of Class I patients treated with nonextraction (NE), all first premolar extractions (PME), and mandibular incisor extraction (MIE) as assessed by the percentage of improvement in Peer Assessment Rating (PAR) scores. This retrospective cross-sectional study was conducted on the pretreatment and posttreatment dental casts of 108 subjects with Class I malocclusion. The total sample was divided into 3 equal groups according to the treatment protocol implemented: NE, PME, and MIE. The mean pretreatment and posttreatment PAR scores, and the percentages of improvement were compared among the 3 treatment modalities using Kruskal-Wallis and post-hoc Dunnett T3 tests. The mean percentages of improvement in the PAR score were 75.8% ± 25.8% in the NE group, 73.1% ± 19.4% in the PME group, and 70.6% ± 24.1% in the MIE group. There was no significant difference (P = 0.351) in the percentages of improvement in PAR scores among the 3 treatment modalities. However, the mean pretreatment and posttreatment PAR scores varied significantly (P <0.001) in the 3 groups. The average pretreatment and posttreatment PAR scores were highest in the MIE group and lowest in the NE group. The comparable percentages of improvement in PAR scores among the 3 groups denote that equivalent occlusal corrections were achieved in Class I patients treated with the NE, PME, and MIE protocols.


Podoplanin expression by cancer-associated fibroblasts predicts poor outcome in invasive ductal breast carcinoma.
It has recently been shown that podoplanin, a mucin-type glycoprotein, is expressed by cancer cells and cancer-associated fibroblasts (CAFs), and promotes cancer cell migration and invasiveness. The biological role of podoplanin expression in tumour stroma of invasive ductal carcinoma of the breast (IDC) has not been determined. Podoplanin expression was analysed in 117 cases of IDC and 27 cases of fibrocystic change, as well as in breast cancer cell lines, with the use of immunohistochemistry and real-time polymerase chain reaction. In 82.1% of analysed tumours, podoplanin was found only in CAFs. Only two of 117 IDC cases (1.7%) were characterized by expression of this glycoprotein in cancer cells. None of the fibrocystic changes or stroma surrounding normal ducts showed podoplanin expression. Podoplanin-positive CAFs correlated with tumour size (P = 0.0125), grade of malignancy (P = 0.0058), lymph node metastasis (P = 0.0149), lymphovascular invasion (LVI) (P = 0.0486) and Ki67 expression in cancer cells (P = 0.0128). High-level podoplanin expression (>50% of positive stroma) in the tumour stroma was significantly associated with a negative oestrogen status (P = 0.0201). Univariate, but not multivariate, analysis showed that podoplanin expression by CAFs was associated with poor patient outcome (P = 0.0202). Our results suggest that podoplanin expression by CAFs could be an unfavourable prognostic marker for IDC.


One Health in Practice: Using Integrated Bite Case Management to Increase Detection of Rabid Animals in Tanzania.
Rabies is a neglected zoonotic disease that causes an estimated 59,000 human deaths worldwide annually, mostly in Africa and Asia. A target of zero human deaths from dog-mediated rabies has been set for 2030, and large-scale control programs are now advocated. However, in most low-income endemic countries surveillance to guide rabies control is weak and few cases of rabies are recorded. There is an urgent need to enhance surveillance to improve timely case detection and inform rabies control and prevention, by operationalizing a "One Health" approach. Here we present data from a study piloting Integrated Bite Case Management (IBCM) to support intersectoral collaboration between health and veterinary workers in Tanzania. We trained government staff to implement IBCM, comprising risk assessments of bite patients by health workers, investigations by livestock field officers to diagnose rabid animals, and use of a mobile phone application to support integration. IBCM was introduced across 20 districts in four regions of Tanzania and results reported after 1 year of implementation. Numbers of bite patient presentations to health facilities varied across regions, but following the introduction of IBCM reporting of bite patients at high-risk for rabies more than doubled in all regions. Over 800 high-risk investigations were carried out, with 49% assessed as probable dog rabies cases on the basis of clinical signs, animal outcome, and rapid diagnostic testing. The status of a further 20% of biting animals could not be determined but rabies could not be ruled out. Livestock field officers reported that use of rapid diagnostic tests (RDTs) were useful for confirming rabies occurrence. Overall, our study provides further evidence that IBCM is a practical approach that can improve rabies detection in endemic countries, and be used to monitor the impact of mass dog vaccinations, including potential to verify rabies freedom. However, the main challenges to implementation are limited training of health workers in rabies, perceived burden of real-time recording and limited resources for livestock field officers to undertake investigations. Nonetheless, IBCM dramatically improved case detection and communication between sectors and we recommend further implementation research to establish best practice and applicability to other settings.


Non-seminomatous germ cell tumor with bone metastasis only at diagnosis: A rare clinical presentation.
Bone metastasis of non-seminomatous germ cell tumors (NSGCT) of the testes is a rare event and even more uncommon at initial presentation. Generally, bone lesions are discovered in the presence of concurrent retroperitoneal lymph node or visceral disease. However, in this case, a 37 years old male complaining of a growing testicular mass was found to have isolated bone metastasis with associated caudaequina syndrome without apparent abnormal findings on initial computed tomography (CT) scans. Continued neurologic symptoms prompted further evaluation with magnetic resonance imaging (MRI), which demonstrated multiple sites of bone metastasis without evidence of retroperitoneal lymph node or visceral organ involvement. This case represents a rare clinical presentation and disease manifestation of NSGCT.


Calcium phosphate nanocapsule crowned multiwalled carbon nanotubes for pH triggered intracellular anticancer drug release.
We report calcium phosphate (CaP) nanocapsule crowned multiwalled carbon nanotubes (CNT-GSH-G4-CaP) as a novel platform for intracellular delivery of an anticancer drug. As a proof-of-concept, CNT-GSH-G4-CaP demonstrates release of anticancer drug doxorubicin hydrochloride (DOX) within intracellular lysosomes from the interior cavity of CNT upon pH triggered CaP dissolution. Importantly, we found that the CNT with a CaP nanolid can efficiently prevent untimely drug release at physiological pH but promotes DOX release at increased acidic milieu as observed in subcellular compartments such as lysosomes (∼5.0). This "zero premature release" characteristic is of clinical significance in delivering cytotoxic drugs, by reducing systemic toxicity and thus beneficial for the effective anticancer treatment. We envision that this pH triggered CaP crowned CNT nanosystem would lead to a new generation of self-regulated platforms for intracellular delivery of a variety of anticancer drugs.


Psychosocial treatment programs for people with both severe mental illness and substance misuse.
Over 50% of people with a severe mental illness also use illicit drugs and/or alcohol at hazardous levels. This review is based on the findings of 25 randomized controlled trials which assessed the effectiveness of psychosocial interventions, offered either as one-off treatments or as an integrated or nonintegrated program, to reduce substance use by people with a severe mental illness. The findings showed that there was no consistent evidence to support any one psychosocial treatment over another. Differences across trials with regard to outcome measures, sample characteristics, type of mental illness and substance used, settings, levels of adherence to treatment guidelines, and standard care all made pooling results difficult. More quality trials are required that adhere to proper randomization methods; use clinically valuable, reliable, and validated measurement scales; and clearly report data, including retention in treatment, relapse, and abstinence rates. Future trials of this quality will allow a more thorough assessment of the efficacy of psychosocial interventions for reducing substance use in this challenging population.


Biochemical properties of amylase isozymes from Gammarus palustris. A comparative study.
Two major alpha-amylase isozymes from Gammarus palustris were purified and characterized. These isozymes, Amy IW and Amy Ic, exhibit a seasonal pattern of expression. In this article we investigate whether the seasonal variation has an adaptive significance. In addition, the species-specific properties of amylases were studied. Purification of the isozymes was achieved by sequential glycogen-ethanol precipitation, Sephadex G-200 and ion exchange chromatography. Characterization in terms of Km, activity patterns at different temperatures, pH values and salt concentrations was done for both isozymes. In addition, the distribution of enzymatic products was analyzed in a high-performance liquid chromatography system. In all conditions tested, the two isozymes gave similar results. This observation suggests that the seasonal change in amylase expression pattern does not result in enzymes differentially suited for seasonal variation in conditions. A comparative analysis showed that G. palustris amylases are apparently distinct from other amylases. These distinctions were seen as an unusually low Km value, an activity peak at low NaCl concentration, a relatively high pH optimum and the predominant formation of maltotriose.


Length heteroplasmies in human mitochondrial DNA control regions and breast cancer risk.
It has been proposed that the presence of heteroplasmy in the hypervariable (HV) regions of the mitochondrial DNA (mtDNA) may be an indicator of mitochondrial genome instability, mtDNA dysfunction, and, thus, may be associated with increased cancer risk. However, whether heteroplasmy in the HV regions of mtDNA could be a risk predictor of oxidative stress-related human cancers, such as breast cancer, remains to be determined. To explore the role of heteroplasmy in the HV regions of mtDNA in breast cancer etiology, we analyzed heteroplasmy in the HV regions of mtDNA in whole blood from 103 patients with breast cancer and 103 matched control subjects. Both cases and controls displayed heteroplasmies in both of the HV1 and HV2 regions. Closer examination of the prevalence of length heteroplasmy indicated that the prevalence of heteroplasmies in both of the HV1 and HV2 regions was much higher in the cases than in the controls (HV1: 68% vs 49%, P=0.007; HV2: 46% vs 25%, P=0.002). The presence of length heteroplasmies in both of the HV1 and HV2 regions was associated with 2.18- and 2.49-folds increased risk of breast cancer, respectively, (HV1: OR=2.18, 95% CI: 1.19 - 4.00; HV2: OR=2.49, 95% CI: 1.32 - 4.69). Interestingly, we observed that the controls with length heteroplasmies in both HV1 and HV2 had statistically significantly lower copy number of mtDNA than the ones without heteroplasmies. These results suggest that the length heteroplasmy in the HV regions of mtDNA could be associated with a risk of breast cancer, perhaps through affecting the copy number of mtDNA.


Subfractionation of hepatic endosomes in Nycodenz gradients and by free-flow electrophoresis. Separation of ligand-transporting and receptor-enriched membranes.
The complexity of rat liver endosome fractions containing internalized radioiodinated asialotransferrin, asialo-(alkaline phosphatase), insulin and prolactin was investigated by using free-flow electrophoresis and isopycnic centrifugation in Nycodenz gradients. Two subfractions were separated by free-flow electrophoresis. Both subfractions contained receptors for asialoglycoprotein and insulin. Glycosyltransferase activities were associated with the more electronegative vesicles, whereas 5'-nucleotidase and alkaline phosphodiesterase activities were associated with the less electronegative vesicles. Three subfractions were separated on Nycodenz gradients. Two subfractions, previously shown to become acidified in vitro, contained the ligands. At short intervals after uptake (1-2 min), ligands were mainly in subfraction DN-2 (density 1.115 g/cm3), but movement into subfraction DN-1 (density 1.090 g/cm3) had occurred 10-15 min after internalization. Low amounts of glycosyltransferase activities were associated with subfraction DN-2, and 5'-nucleotidase and alkaline phosphodiesterase activities were mainly located in subfraction DN-1. The binding sites for asialoglycoproteins and insulin were distributed towards the higher density range in the Nycodenz gradients, thus indicating a segregation of receptor-enriched vesicles and those vesicles containing the various ligands 10-15 min after internalization. Electron microscopy of the subfractions separated on Nycodenz gradients indicated that whereas the ligand-transporting fractions consisted mainly of empty vesicles (average diameter 100-150 nm), the receptor-enriched component was more granular and smaller (average diameter 70-95 nm). The properties of the endosome subfraction are used to assign their origin to the regions of the endocytic compartment where ligand-receptor dissociation and separation occur.


Epiluminescence microscopy for the diagnosis of doubtful melanocytic skin lesions. Comparison of the ABCD rule of dermatoscopy and a new 7-point checklist based on pattern analysis.
To compare the reliability of a new 7-point checklist based on simplified epiluminescence microscopy (ELM) pattern analysis with the ABCD rule of dermatoscopy and standard pattern analysis for the diagnosis of clinically doubtful melanocytic skin lesions. In a blind study, ELM images of 342 histologically proven melanocytic skin lesions were evaluated for the presence of 7 standard criteria that we called the "ELM 7-point checklist." For each lesion, "overall" and "ABCD scored" diagnoses were recorded. From a training set of 57 melanomas and 139 atypical nonmelanomas, odds ratios were calculated to create a simple diagnostic model based on identification of major and minor criteria for the "7-point scored" diagnosis. A test set of 60 melanomas and 86 atypical nonmelanomas was used for model validation and was then presented to 2 less experienced ELM observers, who recorded the ABCD and 7-point scored diagnoses. University medical centers. A sample of patients with excised melanocytic lesions. Sensitivity, specificity, and accuracy of the models for diagnosing melanoma. From the total combined sets, the 7-point checklist gave a sensitivity of 95% and a sepcificity of 75% compared with 85% sensitivity and 66% specificity using the ABCD rule and 91% sensitivity and 90% specificity using standard pattern analysis (overall ELM diagnosis). Compared with the ABCD rule, the 7-point method allowed less experienced observers to obtain higher diagnostic accuracy values. The ELM 7-point checklist provides a simplification of standard pattern analysis because of the low number of features to identify and the scoring diagnostic system. As with the ABCD rule, it can be easily learned and easily applied and has proven to be reliable in diagnosing melanoma.


Onset of Propene Oligomerization Reactivity in ZSM-5 Studied by Inelastic Neutron Scattering Spectroscopy.
The techniques of quasi-elastic and inelastic neutron scattering (QENS and INS) are applied to investigate the oligomerization of propene over a ZSM-5 zeolite. Investigations are performed at low temperatures, allowing identification of the onset of the oligomerization reaction and observation of the low-energy spectral changes due to intermediate formation that are difficult to observe by optical methods. Oligomerization proceeds via formation of a hydrogen-bonded precursor by an interaction of the propene with an internal acid site followed by protonation and chain growth with protonation being the rate-limiting step. The use of quasi-elastic neutron scattering to observe changes in system mobility with temperature via the elastic window scan technique allows identification of the active temperature range where catalyst activity commences and permits targeting of the more time-consuming INS investigations to conditions of interest. From examination of the product's spectrum, the structure of the resulting oligomer is deduced to be primarily linear.


The gas phase structure of coulombically stretched polyethylene glycol ions.
Prior ion-mobility mass-spectrometry (IMS-MS) studies of polyethylene glycol (PEG) ions have identified only two out of many sharply different observed structures: Linear shapes with several individually solvated singly charged cations at high charge states z (beads on a string), and single multiply charged globules at low z. The present study is devoted to assign all other existing structures of PEG ions, for the first time reaching masses of 100 kDa and charge states up to z = 10. There are at most z different structures at charge state z. All involve a single globule carrying n charges, tied to one or several appendices bearing z - n separate charges in a beads-on-a-string configuration. All sharp shape transitions observed at decreasing ion mass involve ejection of one elementary charge (sometimes two) from the shrinking globule into the growing linear appendage. This picture is supported by molecular dynamics simulations and approximate calculations of electrical mobilities for computed structures.


Dietary n-6 PUFA, carbohydrate:protein ratio and change in body weight and waist circumference: a follow-up study.
To investigate the association between the intake of n-6 PUFA and subsequent change in body weight and waist circumference at different levels of the carbohydrate:protein ratio. Follow-up study with anthropometric measurements at recruitment and on average 5·3 years later. Dietary intake was determined at recruitment by using an FFQ that was designed for the study and validated. We applied linear regression models with 5-year change in weight or waist circumference as outcome and including a two-way interaction term between n-6 PUFA and carbohydrate intakes, lower-order terms, protein intake, long-chain n-3 PUFA intake and other potential confounders. Due to adjustment for intake of protein, levels of carbohydrate indirectly reflect levels of the carbohydrate:protein ratio. Diet, Cancer and Health follow-up study, Denmark. Women and men (n 29 152) aged 55 years. For a high intake of n-6 PUFA (6·9 % of energy) v. a low intake of n-6 PUFA (3·4 % of energy), the difference in 5-year weight change was -189·7 g (95 % CI -636·8, 257·4 g) at a low carbohydrate:protein ratio and -86·7 g (95 % CI -502·9, 329·6 g) at a high carbohydrate:protein ratio; the differences in 5-year waist circumference change were 0·26 cm (95 % CI -0·47, 0·98 cm) and -0·52 cm (95 % CI -1·19, 0·15 cm), respectively. Inclusion of the dietary glycaemic index did not change the results. No consistent associations between the intake of n-6 PUFA and change in body weight or waist circumference at different levels of the carbohydrate:protein ratio were observed.


Induction of apoptosis by cytoplasmically localized wild-type p53 and the S121F mutant super p53.
After DNA damage, p53 is accumulated in the nucleus and transactivates downstream genes and induces apoptosis. There are two pathways in p53-dependent apoptosis, the transactivation-dependent and -independent pathway. In this study, we constructed p53-inducible glioblastoma cell lines and analyzed them for the induction of apoptosis and transactivation of p53-downstream genes after the nuclear or cytoplasmic expression of p53. To sequester p53 in the cytoplasm, we used p53 mutant with arginine to glycine substitution at residue 306 (R306G). Wild-type p53 retained the ability to arrest the cell cycle, and a p53 mutant with serine to phenylalanine substitution at residue 121 (S121F), which has a strong ability to induce apoptosis, retained this ability even when both the wild-type and p53 and S121F mutant were exclusively sequestered from the nucleus into the cytoplasm. Notably, cytoplasmically sequestered wild-type p53 and S121F mutant transactivated the downstream genes with distinct expression profiles, and the strong apoptotic ability of S121F was not associated with its transactivation activity. These results underscore the existence of transactivation-independent apoptosis and cytoplasmic function of p53.


Plasma homocyst(e)ine concentrations in eclamptic and preeclamptic African women postpartum.
To examine the relationship between plasma homocyst(e)ine and risk of eclampsia and preeclampsia among sub-Saharan African women who delivered at Harare Maternity Hospital in Zimbabwe. We ran a hospital-based, case-control study at Harare Maternity Hospital, University of Zimbabwe, Harare, Zimbabwe comprising 33 pregnant women with eclampsia and 138 with preeclampsia. Controls were 185 normotensive pregnant women. Plasma was collected postpartum and homocyst(e)ine levels were measured by high-performance liquid chromatography and electrochemical detection. Women with eclampsia or preeclampsia had significantly higher mean homocyst(e)ine levels than normotensive controls (12.54 or 12.77 micromol/L versus 9.93 micromol/L, respectively, P<.001). The odds ratio (OR) for eclampsia was 6.03 among women in the highest quartile of the control homocyst(e)ine distribution (median 13.9 micromol/L) compared with women in the lowest quartile (median 6.2 micromol/L). The corresponding OR for preeclampsia was 4.57. Nulliparas with elevated homocyst(e)ine had a 12.90 times higher risk of preeclampsia compared with multiparas without elevated homocyst(e)ine. Postpartum plasma homocyst(e)ine concentrations are higher among Zimbabwean women with eclampsia and preeclampsia compared with normotensive women.


Clinical elements for the neuromuscular stimulation and functional electrical stimulation protocols in the practice of neurorehabilitation.
The physicians and their multidisciplinary teams involved in the clinical practice of neurological rehabilitation have more and more opportunities to apply neuromuscular stimulation (NMS) and functional electrical stimulation (FES) of peripheral nerves as a part of their daily practice. In this article, we outline clinical protocols of NMS and FES in the following clinical conditions of upper motor neuron dysfunction: to prevent consequences of disuse of the neuromuscular system of the upper motor neuron, to facilitate recovery processes of impaired upper motor neuron functions due to acute and/or subacute neurological conditions, to maintain or enhance the trophic state of the muscle, to modify altered control of muscle tone, to modify altered patterns of automatic and volitional functional movements, to enhance functional movement of the single joint muscle group within intact functional multijoint movement, and to modify altered neurocontrol of posture, locomotion, and skillful movements. We emphasize the importance of understanding the motor control alteration while developing clinical protocols and defining the goals. It is very important to be aware that similar clinical findings and due to the same cause can have different features of residual motor control, and therefore potentials for recovery or modification can be very different.


Adjustment Disorder and Suicidal Behaviours Presenting in the General Medical Setting: A Systematic Review.
Background: Adjustment disorder (AD) is a condition commonly encountered by clinicians in emergency departments and liaison psychiatry settings and has been frequently reported among patients presenting with suicidal behaviours. A number of previous studies have noted the strong association between suicidal ideation and behaviours, and AD. In this paper, we aimed to explore this relationship, by establishing the incidence of AD in patients who present with self-harm and suicidal ideation, and the rates of self-harm among patients with a diagnosis of AD. Methods: We conducted a review of the literature of well-established databases using specific key words then synthesised the results into a descriptive narrative as well as representing it in table form. Results: Sample sizes and study methods varied significantly across the review. A majority of studies were retrospective chart-based reviews, and only three used structured diagnostic instruments. A high prevalence of AD (ranging from 9.8 to 100%) was found, with self-poisoning representing the most common form of suicide attempt in the majority of studies. Interpersonal difficulties were the main precipitant in studies which examined this. Conclusions: This study suggests there is a strong association between AD and suicidal behaviours. Given the paucity of research in the area, there is a need to build the evidence base for effective treatment strategies.