Contralateral cortico-ponto-cerebellar pathways reconstruction in humans in vivo: implications for reciprocal cerebro-cerebellar structural connectivity in motor and non-motor areas.
Cerebellar involvement in cognition, as well as in sensorimotor control, is increasingly recognized and is thought to depend on connections with the cerebral cortex. Anatomical investigations in animals and post-mortem humans have established that cerebro-cerebellar connections are contralateral to each other and include the cerebello-thalamo-cortical (CTC) and cortico-ponto-cerebellar (CPC) pathways. CTC and CPC characterization in humans in vivo is still challenging. Here advanced tractography was combined with quantitative indices to compare CPC to CTC pathways in healthy subjects. Differently to previous studies, our findings reveal that cerebellar cognitive areas are reached by the largest proportion of the reconstructed CPC, supporting the hypothesis that a CTC-CPC loop provides a substrate for cerebro-cerebellar communication during cognitive processing. Amongst the cerebral areas identified using in vivo tractography, in addition to the cerebral motor cortex, major portions of CPC streamlines leave the prefrontal and temporal cortices. These findings are useful since provide MRI-based indications of possible subtending connectivity and, if confirmed, they are going to be a milestone for instructing computational models of brain function. These results, together with further multi-modal investigations, are warranted to provide important cues on how the cerebro-cerebellar loops operate and on how pathologies involving cerebro-cerebellar connectivity are generated.


Intra-ocular pressure variation associated with the wear of scleral lenses of different diameters.
To evaluate the variation of intra-ocular pressure during scleral lens wear, and the influence of the lens diameter on the results. This is a prospective, randomized study performed on Caucasian subjects (16 F; 5 M), aged 24.7 + 4.1 y.o. A diurnal variation pattern (IOPg) was established, then, transpalpebral IOP (IOPt) was taken before and during SL wear. One eye, randomly fitted with a 15.8 diameter SL (L1), was compared to the fellow eye, fitted with an 18 mm SL of the same design, thickness and material (L2). Anterior segment tomography was taken pre-and after lens removal. Baseline IOPg (L1:15.2 + 3.1 mm HG; L2: 15.1 +/- 2.8 mm) did not reveal significant diurnal variations. Wearing L1, IOPt rose from 10.1 + 1.9 mm HG to 14.4 + 5.5 mm HG after 4.5 + 0.3 hrs, while with L2, it rose from 9.2 + 2.1 mm HG to 14.4 + 4.8 mm Hg. This difference is statistically significant based on time but not on lenses. Anterior segment parameters did not vary except for the anterior chamber volume (L1: -1.53 + 7.61 mm3; L2: -3.47 + 6.4 mm3), and for the corneal thickness (+2.1% with L1 and L2). These results suggest that, as evaluated with a non-standard transpalpebral methodology, IOP during scleral lens wear may be increased in average by 5 mm Hg, regardless of the lens diameter. More work is needed to confirm if practitioners should be warned when using SL on populations at risk for glaucoma.


Modeling coupled kinetics of antimony adsorption/desorption and oxidation on manganese oxides.
Understanding the kinetic reactions of antimony (Sb) at the MnO2-water interface is essential for predicting the dynamic behavior of Sb in soil environments. In this study, we developed a quantitative model for assessing the coupling between Sb(iii) oxidation and Sb(iii)/Sb(v) adsorption/desorption kinetics and the role of each individual reaction process in controlling the overall reaction rates. Based on our modeling results, Sb(iii) oxidation rates were very fast at the early stages of the reactions and may significantly slow down with time, and Sb(iii) and Sb(v) showed different adsorption behavior on δ-MnO2. Our kinetics model is able to account for different adsorption/desorption kinetics of Sb(iii) and Sb(v) and the changes of Sb(iii) oxidation rates during the coupled kinetic processes, which provides a general modeling framework for predicting Sb kinetic behavior at the MnO2-water interface.


Principles and practice of parenteral nutrition in the neonatal period.
In extremely preterm or critically ill infants, the parenteral route for maintaining nutritional integrity has to be relied upon before successful transition to the enteral route of feeding is achieved. Parenteral nutrition is now a fundamental part of neonatal intensive care. Fluid intake volumes vary from 60-150 ml/kg/d, depending on maturity of the infant and environmental conditions influencing insensible water loss from the skin. Parenteral nitrogen requirements are 30-35 mmol/kg/d, equivalent to 3.0-3.5 mg/kg/d of amino acids. Hyperglycaemia during parenteral nutrition can be minimised by starting glucose infusion at a rate of 6-8 g/kg/d with progressive increase to 18-20 g/kg/d by 2-3 weeks after birth. Parenteral fat is introduced at 1 g/kg/d, gradually increasing to 3 g/kg/d, given as a continuous infusion. An energy intake of 50 kcal/kg/d is adequate to match ongoing expenditure but an additional energy intake of 70 kcal/kg/d is required to achieve optimal growth. Minerals and trace elements delivered with parenteral nutrition are calculated to meet in-utero accretion rates. Multivitamins available for parenteral use should also be included. Improved techniques for the preparation, administration and monitoring of parenteral nutrition have helped minimise catheter-related and metabolic complications. In neonatal intensive care units where appropriate medical, nursing, pharmacy and laboratory expertise are available, the potential benefits of parenteral nutrition outweigh its hazards. Nevertheless, early initiation of enteral feeding in small subnutritional quantities to supplement parenteral nutrition is of major importance to enhance the growth and development of the gastrointestinal tract.


Intermediate CD14++CD16+ monocyte predicts severe coronary stenosis and extensive plaque involvement in asymptomatic individuals.
Circulating leukocyte subtypes and monocyte subsets are independent predictors of cardiovascular events. We hypothesized that an increased leukocyte subtype would predict severe coronary stenosis and extensive plaque involvement. We retrospectively analyzed clinical, laboratory, and coronary CT data in a total of 588 asymptomatic adults (69% men; mean age, 57 ± 9 years) undergoing a general health check-up. Intermediate CD14++CD16+ monocyte count had the strongest association with mixed and calcified plaque scores, whereas the numbers of neutrophils and classical CD14++CD16- monocytes were significantly associated with non-calcified plaque score. Only high CD14++CD16+ monocyte count (>12 cells/μL) significantly predicted extensive plaque involvement [odds ratio 3.16 (95% confidence interval 1.84-5.43), P < 0.001; quartile 4 vs. 1-3] and severe coronary stenosis [3.67 (1.84-7.33), P < 0.001; quartile 4 vs. 1-3] after adjustments for Framingham Risk Score (FRS), metabolic syndrome, and C-reactive protein. The CD14++CD16+ monocyte count, when added to FRS, significantly reclassified 30.4 and 26.7% of the overall and 50.2 and 36.2% of the intermediate-risk population (FRS 6-20%) for predicting extensive plaque involvement and severe coronary stenosis, respectively. Thus, in asymptomatic individuals, intermediate CD14++CD16+ monocyte could independently predict severe CAD and improve risk stratification.


Haltere-mediated equilibrium reflexes of the fruit fly, Drosophila melanogaster.
Flies display a sophisticated suite of aerial behaviours that require rapid sensory-motor processing. Like all insects, flight control in flies is mediated in part by motion-sensitive visual interneurons that project to steering motor circuitry within the thorax. Flies, however, possess a unique flight control equilibrium sense that is encoded by mechanoreceptors at the base of the halteres, small dumb-bell-shaped organs derived through evolutionary transformation of the hind wings. To study the input of the haltere system onto the flight control system, I constructed a mechanically oscillating flight arena consisting of a cylindrical array of light-emitting diodes that generated the moving image of a 30 degrees vertical stripe. The arena provided closed-loop visual feedback to elicit fixation behaviour, an orientation response in which flies maintain the position of the stripe in the front portion of their visual field by actively adjusting their wing kinematics. While flies orientate towards the stripe, the entire arena was swung back and forth while an optoelectronic device recorded the compensatory changes in wing stroke amplitude and frequency. In order to reduce the background changes in stroke kinematics resulting from the animal's closed-loop visual fixation behaviour, the responses to eight identical mechanical rotations were averaged in each trial. The results indicate that flies possess a robust equilibrium reflex in which angular rotations of the body elicit compensatory changes in both the amplitude and stroke frequency of the wings. The results of uni- and bilateral ablation experiments demonstrate that the halteres are required for these stability reflexes. The results also confirm that halteres encode angular velocity of the body by detecting the Coriolis forces that result from the linear motion of the haltere within the rotating frame of reference of the fly's thorax. By rotating the flight arena at different orientations, it was possible to construct a complete directional tuning map of the haltere-mediated reflexes. The directional tuning of the reflex is quite linear such that the kinematic responses vary as simple trigonometric functions of stimulus orientation. The reflexes function primarily to stabilize pitch and yaw within the horizontal plane.


Effects of gamma-tocopherol supplementation on thrombotic risk factors.
The antioxidant activity of vitamin E is derived primarily from alpha-tocopherol (alpha-T) and gamma-tocopherol (gamma-T). Results of epidemiological studies have demonstrated an inverse relationship between vitamin E intake and coronary disease. However, the results of clinical trials using alpha-T are equivocal. We determined the effect of 5 weeks of 100 mg/d or 200 mg/d gamma-T supplementation on thrombotic markers such as platelet reactivity, lipid profile and the inflammation marker C-reactive protein (CRP). Fourteen healthy subjects consumed 100 mg/day while 13 consumed 200 mg/d of gamma-T and 12 received placebo (soybean capsules with less than 5 mg/d gamma-T) in a double-blinded parallel study design. Fasting pre and post dose blood samples were analysed. Blood gamma-T concentrations increased significantly (p<0.05) relative to dose during the intervention period. Both groups receiving active ingredients showed significantly lower platelet activation after supplementation (p<0.05). Subjects consuming 100 mg/d gamma-T had significantly decreased LDL cholesterol, platelet aggregation and mean platelet volume (MPV) (p<0.05). Little effect of gamma-T was observed on other parameters. These data suggest that gamma-T supplementation may have a permissive role in decreasing the risk of thrombotic events by improving lipid profile and reducing platelet activity.


Socio-cultural Dimensions of Congenital Adrenal Hyperplasia: An Ethnographic Study from Chennai, South India.
This study aims to provide a medical anthropological perspective on how congenital adrenal hyperplasia (CAH) is perceived and constructed by parents and doctors in India. It aims to put forth the complexities that are associated with CAH and the various experiences that parents and doctors share as a result, while also exploring the influences that culture and medicine have on each other. An ethnographic approach was taken to understand CAH in this study, in which families and doctors of children with CAH were interviewed. Fieldwork was done for 2 months in Chennai, Tamil Nadu. A major finding of this study was the faith that parents had on biomedicine in general and doctors in particular. While parents continued to follow the instructions provided by the doctors, they also exercised their agency by questioning the decisions taken by the doctors. The research also revealed that there is constant worry and fear in parents about the future of their children due to the stigma attached to CAH. A constant discourse between medicine and culture can be noticed while analyzing the complexities associated with CAH. The study tries to show that medical decisions that doctors take in matters concerning CAH are culturally driven. Surgical corrections done in order to categorize the child into one of the two sexes is an example for the same. Similarly, various structures of family, marriage, and kinship have been medicalized owing to the strong influence medicine and culture have on each other.


Indo-European and Asian origins for Chilean and Pacific chickens revealed by mtDNA.
European chickens were introduced into the American continents by the Spanish after their arrival in the 15th century. However, there is ongoing debate as to the presence of pre-Columbian chickens among Amerindians in South America, particularly in relation to Chilean breeds such as the Araucana and Passion Fowl. To understand the origin of these populations, we have generated partial mitochondrial DNA control region sequences from 41 native Chilean specimens and compared them with a previously generated database of approximately 1,000 domestic chicken sequences from across the world as well as published Chilean and Polynesian ancient DNA sequences. The modern Chilean sequences cluster closely with haplotypes predominantly distributed among European, Indian subcontinental, and Southeast Asian chickens, consistent with a European genetic origin. A published, apparently pre-Columbian, Chilean specimen and six pre-European Polynesian specimens also cluster with the same European/Indian subcontinental/Southeast Asian sequences, providing no support for a Polynesian introduction of chickens to South America. In contrast, sequences from two archaeological sites on Easter Island group with an uncommon haplogroup from Indonesia, Japan, and the Philippines [corrected] and may represent a genetic signature of an early Polynesian dispersal. Modeling of the potential marine carbon contribution to the Chilean archaeological specimen casts further doubt on claims for pre-Columbian chickens, and definitive proof will require further analyses of ancient DNA sequences and radiocarbon and stable isotope data from archaeological excavations within both Chile and Polynesia.


Cross-Feeding among Probiotic Bacterial Strains on Prebiotic Inulin Involves the Extracellular exo-Inulinase of Lactobacillus paracasei Strain W20.
Probiotic gut bacteria employ specific metabolic pathways to degrade dietary carbohydrates beyond the capabilities of their human host. Here, we report how individual commercial probiotic strains degrade prebiotic (inulin type) fructans. First, a structural analysis of commercial fructose oligosaccharide-inulin samples was performed. These β-(2-1)-fructans differ in termination by either glucose (GF) or fructose (FF) residues, with a broad variation in the degrees of polymerization (DPs). The growth of individual probiotic bacteria on short-chain inulin (sc-inulin) (Frutafit CLR), a β-(2-1)-fructan (DP 2 to DP 40), was studied. Lactobacillus salivarius W57 and other bacteria grew relatively poorly on sc-inulin, with only fractions of DP 3 and DP 5 utilized, reflecting uptake via specific transport systems followed by intracellular metabolism. Lactobacillus paracasei subsp. paracasei W20 completely used all sc-inulin components, employing an extracellular exo-inulinase enzyme (glycoside hydrolase family GH32 [LpGH32], also found in other strains of this species); the purified enzyme converted high-DP compounds into fructose, sucrose, 1-kestose, and F2 (inulobiose). The cocultivation of L. salivarius W57 and L. paracasei W20 on sc-inulin resulted in cross-feeding of the former by the latter, supported by this extracellular exo-inulinase. The extent of cross-feeding depended on the type of fructan, i.e., the GF type (clearly stimulating) versus the FF type (relatively low stimulus), and on fructan chain length, since relatively low-DP β-(2-1)-fructans contain a relatively high content of GF-type molecules, thus resulting in higher concentrations of GF-type DP 2 to DP 3 degradation products. The results provide an example of how in vivo cross-feeding on prebiotic β-(2-1)-fructans may occur among probiotic lactobacilli.IMPORTANCE The human gut microbial community is associated strongly with host physiology and human diseases. This observation has prompted research on pre- and probiotics, two concepts enabling specific changes in the composition of the human gut microbiome that result in beneficial effects for the host. Here, we show how fructooligosaccharide-inulin prebiotics are fermented by commercial probiotic bacterial strains involving specific sets of enzymes and transporters. Cross-feeding strains such as Lactobacillus paracasei W20 may thus act as keystone strains in the degradation of prebiotic inulin in the human gut, and this strain-exo-inulinase combination may be used in commercial Lactobacillus-inulin synbiotics.


Design of isoform-selective phospholipase D inhibitors that modulate cancer cell invasiveness.
Phospholipase D (PLD) is an essential enzyme responsible for the production of the lipid second messenger phosphatidic acid. Phosphatidic acid participates in both G protein-coupled receptor and receptor tyrosine kinase signal transduction networks. The lack of potent and isoform-selective inhibitors has limited progress in defining the cellular roles of PLD. We used a diversity-oriented synthetic approach and developed a library of PLD inhibitors with considerable pharmacological characterization. Here we report the rigorous evaluation of that library, which contains highly potent inhibitors, including the first isoform-selective PLD inhibitors. Specific members of this series inhibit isoforms with >100-fold selectivity both in vitro and in cells. A subset of inhibitors was shown to block invasiveness in metastatic breast cancer models. These findings demonstrate the power of diversity-oriented synthesis combined with biochemical assays and mass spectrometric lipid profiling of cellular responses to develop the first isoform-selective PLD inhibitors--a new class of antimetastatic agents.


Effect of auditory input on activations in infant diverse cortical regions during audiovisual processing.
A fundamental question with regard to perceptual development is how multisensory information is processed in the brain during the early stages of development. Although a growing body of evidence has shown the early emergence of modality-specific functional differentiation of the cortical regions, the interplay between sensory inputs from different modalities in the developing brain is not well understood. To study the effects of auditory input during audio-visual processing in 3-month-old infants, we evaluated the spatiotemporal cortical hemodynamic responses of 50 infants while they perceived visual objects with or without accompanying sounds. The responses were measured using 94-channel near-infrared spectroscopy over the occipital, temporal, and frontal cortices. The effects of sound manipulation were pervasive throughout the diverse cortical regions and were specific to each cortical region. Visual stimuli co-occurring with sound induced the early-onset activation of the early auditory region, followed by activation of the other regions. Removal of the sound stimulus resulted in focal deactivation in the auditory regions and reduced activation in the early visual region, the association region of the temporal and parietal cortices, and the anterior prefrontal regions, suggesting multisensory interplay. In contrast, equivalent activations were observed in the lateral occipital and lateral prefrontal regions, regardless of sound manipulation. Our findings indicate that auditory input did not generally enhance overall activation in relation to visual perception, but rather induced specific changes in each cortical region. The present study implies that 3-month-old infants may perceive audio-visual multisensory inputs by using the global network of functionally differentiated cortical regions.


Early-life seizures result in deficits in social behavior and learning.
Children with epilepsy show a high co-morbidity with psychiatric disorders and autism. One of the critical determinants of a child's behavioral outcome with autism and cognitive dysfunction is the age of onset of seizures. In order to examine whether seizures during postnatal days 7-11 result in learning and memory deficits and behavioral features of autism we administered the inhalant flurothyl to induce seizures in C57BL/6J mice. Mice received three seizures per day for five days starting on postnatal day 7. Parallel control groups consisted of similarly handled animals that were not exposed to flurothyl and naïve mice. Subjects were then processed through a battery of behavioral tests in adulthood: elevated-plus maze, nose-poke assay, marble burying, social partition, social chamber, fear conditioning, and Morris water maze. Mice with early-life seizures had learning and memory deficits in the training portion of the Morris water maze (p<0.05) and probe trial (p<0.01). Mice with seizures showed no differences in marble burying, the nose-poke assay, or elevated plus-maze testing compared to controls. However, they showed a significant difference in the social chamber and social partition tests. Mice with seizures during postnatal days 7-11 showed a significant decrease in social interaction in the social chamber test and had a significant impairment in social behavior in the social partition test. Together, these results indicate that early life seizures result in deficits in hippocampal-dependent memory tasks and produce long-term disruptions in social behavior.


One year followup variables predict disability 5 years after presentation with inflammatory polyarthritis with greater accuracy than at baseline.
To compare the accuracy of simple demographic and clinical variables recorded at baseline with those recorded after one year followup, in predicting self-reported functional disability recorded 5 years after initial assessment in patients with early inflammatory polyarthritis (IP). We followed annually for 5 years 528 patients registered by the Norfolk Arthritis Register (a primary care based cohort of patients with early IP) using the Health Assessment Questionnaire (HAQ). Backward stepwise logistic regression was used to determine the clinical and demographic variables, collected at either baseline or first followup, that were associated with disability (HAQ > or = 1.00/> or =1.50) at 5 years. At the 5th anniversary assessment, the prevalence of moderate disability (HAQ > or = 1.00) was 47%. Twenty-nine percent reported more severe disability (HAQ > or = 1.50). Variables recorded at first anniversary assessment were better able to predict patients at risk of developing a poor outcome than baseline variables. Multivariate methods identified age at symptom onset, HAQ score, presence of nodules, and a statistically derived factor describing joint tenderness recorded at first year as important predictors of both moderate disability (HAQ > or =1.00) and a higher level of disability (HAQ > or = 1.50). When tested in an independent validation sample, the accuracy of the models generated from data recorded at the first year was 76% (HAQ > or = 1.00) and 83% (HAQ > or = 1.50). It was possible to predict disability at 5 years with high accuracy using simple clinical variables and demographic data collected 4 or 5 years previously. First year HAQ score was the strongest predictor of future disability. HAQ score at 5 years could be predicted more accurately using data collected at first anniversary visit than using data recorded at baseline.


New Features for Measuring Disease Activity in Pediatric Localized Scleroderma.
To identify clinical features that define disease activity in pediatric localized scleroderma (LS), and determine their specificity and importance. We conducted a multicenter prospective study of patients with active and inactive LS skin lesions. A standardized evaluation of a single designated study lesion per subject was performed at 3 visits. We evaluated the pattern and correlation between assessed features and physician's global assessments of activity (PGA-A). Ninety of 103 subjects had evaluable data; 66 had active and 24 inactive disease. Subjects had similar age of onset, sex, and disease patterns. Linear scleroderma was the most common subtype. Features specific for active disease included erythema, violaceous color, tactile warmth, abnormal skin texture, and disease extension. Scores for these variables changed over time and correlated with PGA-A of the lesion. Active and inactive lesions could not be distinguished by the presence or level of skin thickening, either of lesion edge or center. However, in active lesions, skin thickening scores did correlate with PGA-A scores. Regression analysis identified the combination of erythema, disease extension, violaceous color, skin thickening, and abnormal texture as predictive of PGA-A at study entry. Damage features were common irrespective of activity status. We identified variables strongly associated with disease activity, expanding upon those used in current measures, and determined their relative importance in physician activity scoring. Skin thickening was found to lack specificity for disease activity. These results will help guide development of a sensitive, responsive activity tool to improve care of patients with LS.


Spinal instrumentation for sacral-pelvic fixation: a biomechanical comparison between constructs ending with either S2 bicortical, bitriangulated screws or iliac screws.
A biomechanical study of 2 fixation techniques for lumbosacral fixation. To evaluate 2 techniques, one using S1 screws combined with bicortical, bitriangulated (BCBT) S2 screws, and the other using S1 screws combined with iliac screws. Common to the 2 techniques of sacral-pelvic fixation is S1 pedicle screws; the difference lies between S2 screws versus iliac screws. Iliac screws are clinically effective, yet present clinical disadvantages that S2 screws can potentially obviate, for example, wide dissection, soft tissue coverage, crossing the sacroiliac joint, and interference with bone graft harvesting. In an effort to optimize S2 fixation, we have used a BCBT S2 technique. Eight fresh human sacral-pelvic specimens were harvested (average age 78.7 y; bone density 0.75 g/cm2). Screws were placed bilaterally: (1) at S1: 7.5 mm diameter screws were placed bicortical; (2) at S2: 7.5 mm diameter by 60 mm long screws were placed bicortical and bitriangulated; (3) the ilium received 7.5 mm diameter by 80-mm-long screws. Sacral-pelvic constructs were assembled and biomechanical stiffness testing was performed. The stiffness in each loading mode was calculated. After the stiffness tests were completed, each BCBT S2 screw and each iliac screw were individually loaded to determine fixation strength of each type of screw. There was no significant difference in stiffness between the 2 constructs, although S1 and the BCBT S2 construct tended to be stiffer in all modes. However, for the fixation strength of screws, the iliac screws loosened at loads that were significantly greater than those for BCBT S2 screws (c.f. 435.9 N with 144.7 N). Iliac screws maybe the better choice in cases with poor bone quality. However, the S1-BCBT-S2 screw construct is biomechanically an appropriate alternative to the S1-iliac screw construct and it presents clinical advantages.


Immunoperoxidase detection of polycyclic aromatic hydrocarbon-DNA adducts in oral mucosa cells of smokers and nonsmokers.
An immunoperoxidase method using a polyclonal antiserum which recognizes benzo(a)pyrene and structurally related polycyclic aromatic hydrocarbon diol epoxide-DNA adducts has been developed for the detection and quantitation of DNA damage in single cells. The method was used initially on 10T1/2 cells treated with [3H]anti-benzo(a)pyrene diol epoxide then applied to the detection of adducts in oral mucosa cells of smokers and nonsmokers. Levels of DNA damage were elevated in each of 16 smokers (mean relative staining, 503 +/- 104) compared to 16 age-, race- and sex-matched nonsmokers (251 +/- 82; P < 0.0001). There was an approximately 3-fold range in relative staining in both smokers (252 +/- 125 to 663 +/- 189) and nonsmokers (157 +/- 72 to 431 +/- 269) suggesting the importance of individual differences in capacity to metabolize the carcinogens and/or repair damaged DNA. This noninvasive method, requiring small numbers of cells, will be useful for routine monitoring of DNA damage in intervention studies as well as for biofeedback in smoking cessation programs.


Application of machine learning-based models to boost the predictive power of the SPAN index.
This study re-explored the predictive validity of Stroke Prognostication using Age and National Institutes of Health Stroke Scale (SPAN) index in patients who received different treatments for acute ischemic stroke (AIS) and developed machine learning-boosted outcome prediction models. We evaluated the prognostic relevance of SPAN index in patients with AIS who received intravenous tissue-type plasminogen activator (IV-tPA), intra-arterial thrombolysis (IAT) or non-thrombolytic treatments (non-tPA), and applied machine learning algorithms to develop SPAN-based outcome prediction models in a cohort of 2145 hospitalized AIS patients. The performance of the models was assessed and compared using the area under the receiver operating characteristic curves (AUCs). SPAN index ≥100 was associated with higher mortality rate and higher modified Rankin Scale at discharge in AIS patients who received the different treatments. Compared to the lower AUCs for the SPAN-alone model across all groups, the AUCs of the logistic regression-boosted model were 0.838, 0.857, 0.766 and 0.875 for the whole cohort, non-tPA, IV-tPA and IAT groups, respectively. Similarly, the AUCs of the generated artificial neural network were 0.846, 0.858, 0.785 and 0.859 for the whole cohort, non-tPA, IV-tPA and IAT groups, respectively, while for gradient boosting decision tree model, we computed 0.850, 0.863, 0.779 and 0.815. SPAN index has prognostic relevance in patients with AIS who received different treatments. The generated machine learning-based models exhibit good performance for predicting the functional recovery of AIS; thus, their proposed clinical application to aid outcome prediction and decision-making for the patients with AIS.


Human carrying simulation with symmetric and asymmetric loads using optimization.
Human carrying is simulated in this work by using a skeletal digital human model with 55 degrees of freedom. An optimization-based approach is used to predict the carrying motion with symmetric and asymmetric loads. In this process, the model predicts joint dynamics using optimization schemes and task-based physical constraints. The results indicate that the model can predict different carrying strategies during symmetric and asymmetric load-carrying tasks. The model can also indicate the risk factors for extreme loading situations. With such robust prediction capability, the model could be used for biomedical and ergonomic studies.


In vivo administration of a lentiviral vaccine targets DCs and induces efficient CD8(+) T cell responses.
The present study evaluates the potential of third-generation lentivirus vectors with respect to their use as in vivo-administered T cell vaccines. We demonstrate that lentivector injection into the footpad of mice transduces DCs that appear in the draining lymph node and in the spleen. In addition, a lentivector vaccine bearing a T cell antigen induced very strong systemic antigen-specific cytotoxic T lymphocyte (CTL) responses in mice. Comparative vaccination performed in two different antigen models demonstrated that in vivo administration of lentivector was superior to transfer of transduced DCs or peptide/adjuvant vaccination in terms of both amplitude and longevity of the CTL response. Our data suggest that a decisive factor for efficient T cell priming by lentivector might be the targeting of DCs in situ and their subsequent migration to secondary lymphoid organs. The combination of performance, ease of application, and absence of pre-existing immunity in humans make lentivector-based vaccines an attractive candidate for cancer immunotherapy.


Understanding Articular Cartilage Injury and Potential Treatments.
The goals of all orthopaedic surgeons treating articular cartilage injuries have been anatomic reduction and stable fixation of the articular cartilage surface with restoration of limb alignment and/or reestablishment of the joint stability, all while minimizing the risk of surgical complications. Recent developments in the study of articular cartilage injury have shown that there is a robust cellular response to joint injury. This response has been shown to involve the synoviocytes, chondrocytes, and osteocytes in and around the injured joint and if these responses are left unchecked, they can lead to the development of posttraumatic osteoarthritis (PTOA). Therefore, to predictably and successfully treat articular cartilage injuries, it is not sufficient to just restore articular congruity, limb alignment, and joint stability, but we must also recognize and attempt to mitigate this associated cellular response. Understanding not only the mechanical aspects of these joint injuries but also the biological aspects is paramount to giving our patients the best opportunity to heal their injuries, recover full function, and avoid the potential devastating development of PTOA. Gone is the simplistic view that if one can achieve articular congruity after intraarticular fracture, as well as joint stability after ligamentous injury, that our patients will do just fine. This review sheds new light on the molecular response to cartilage injury, how residual joint incongruity and instability affect the joint's ability to recover from injury, and how chondrocyte apoptosis in response to injury can influence joint. This article then briefly reviews how cellular and growth factors may be beneficial to the treatment of articular cartilage injury and how ultimately cartilage regeneration may be used in the future to salvage the joints ravaged by PTOA in response to injury.


FastGGM: An Efficient Algorithm for the Inference of Gaussian Graphical Model in Biological Networks.
Biological networks provide additional information for the analysis of human diseases, beyond the traditional analysis that focuses on single variables. Gaussian graphical model (GGM), a probability model that characterizes the conditional dependence structure of a set of random variables by a graph, has wide applications in the analysis of biological networks, such as inferring interaction or comparing differential networks. However, existing approaches are either not statistically rigorous or are inefficient for high-dimensional data that include tens of thousands of variables for making inference. In this study, we propose an efficient algorithm to implement the estimation of GGM and obtain p-value and confidence interval for each edge in the graph, based on a recent proposal by Ren et al., 2015. Through simulation studies, we demonstrate that the algorithm is faster by several orders of magnitude than the current implemented algorithm for Ren et al. without losing any accuracy. Then, we apply our algorithm to two real data sets: transcriptomic data from a study of childhood asthma and proteomic data from a study of Alzheimer's disease. We estimate the global gene or protein interaction networks for the disease and healthy samples. The resulting networks reveal interesting interactions and the differential networks between cases and controls show functional relevance to the diseases. In conclusion, we provide a computationally fast algorithm to implement a statistically sound procedure for constructing Gaussian graphical model and making inference with high-dimensional biological data. The algorithm has been implemented in an R package named "FastGGM".


Persistent infection or successive reinfection of deer mice with Bartonella vinsonii subsp. arupensis.
Bartonella infections are common in rodents. From 1994 to 2006, longitudinal studies of a rodent community, consisting mainly of deer mice (Peromyscus maniculatus), were conducted in southwestern Colorado to study hantaviruses. Blood samples from deer mice captured one or more times during the period 2003 to 2006 (n = 737) were selected to study bartonellae in deer mice. Bartonellae were found to be widely distributed in that population, with an overall prevalence of 82.4% (607/737 mice). No correlation was found between bartonella prevalence and deer mouse weight or sex. Persistent or successive infections with bartonellae were observed in deer mice captured repeatedly, with a prevalence of 83.9% (297/354), and the infection appeared to last for more than 1 year in some of them. Persistent infection with bartonellae may explain the high prevalence of these bacteria in deer mice at this site and, perhaps, elsewhere. Genetic analysis demonstrated that deer mouse-borne bartonella isolates at this site belong to the same species, B. vinsonii subsp. arupensis, demonstrating a specific relationship between B. vinsonii subsp. arupensis and deer mice.


iCLIP--transcriptome-wide mapping of protein-RNA interactions with individual nucleotide resolution.
The unique composition and spatial arrangement of RNA-binding proteins (RBPs) on a transcript guide the diverse aspects of post-transcriptional regulation. Therefore, an essential step towards understanding transcript regulation at the molecular level is to gain positional information on the binding sites of RBPs. Protein-RNA interactions can be studied using biochemical methods, but these approaches do not address RNA binding in its native cellular context. Initial attempts to study protein-RNA complexes in their cellular environment employed affinity purification or immunoprecipitation combined with differential display or microarray analysis (RIP-CHIP). These approaches were prone to identifying indirect or non-physiological interactions. In order to increase the specificity and positional resolution, a strategy referred to as CLIP (UV cross-linking and immunoprecipitation) was introduced. CLIP combines UV cross-linking of proteins and RNA molecules with rigorous purification schemes including denaturing polyacrylamide gel electrophoresis. In combination with high-throughput sequencing technologies, CLIP has proven as a powerful tool to study protein-RNA interactions on a genome-wide scale (referred to as HITS-CLIP or CLIP-seq). Recently, PAR-CLIP was introduced that uses photoreactive ribonucleoside analogs for cross-linking. Despite the high specificity of the obtained data, CLIP experiments often generate cDNA libraries of limited sequence complexity. This is partly due to the restricted amount of co-purified RNA and the two inefficient RNA ligation reactions required for library preparation. In addition, primer extension assays indicated that many cDNAs truncate prematurely at the crosslinked nucleotide. Such truncated cDNAs are lost during the standard CLIP library preparation protocol. We recently developed iCLIP (individual-nucleotide resolution CLIP), which captures the truncated cDNAs by replacing one of the inefficient intermolecular RNA ligation steps with a more efficient intramolecular cDNA circularization (Figure 1). Importantly, sequencing the truncated cDNAs provides insights into the position of the cross-link site at nucleotide resolution. We successfully applied iCLIP to study hnRNP C particle organization on a genome-wide scale and assess its role in splicing regulation.


Sensitivity of FRAMINGHAM, PROCAM and SCORE models in Turkish people with Type 2 diabetes: comparison of three cardiovascular risk calculations.
To determine the cardiovascular risk factors according to the Framingham, PROCAM and SCORE models, to evaluate the 10-year cardiovascular disease (CVD) risk factors, and to compare the suitability of different models in Turkish Type 2 diabetes patients. Risk factors and the 10-year CVD risk in 265 patients were evaluated using three risk models. Measurements included blood pressure, weight, height, waist and hip circumferences. Cholesterol, triglycerides, fasting and postprandial plasma glucose and HbA1c were measured. Low-, moderate- and high-risk groups were determined according to the three risk calculations. Hypertension, obesity and no exercise, dyslipidemia and high HbA1c in women, and excessive cigarette/alcohol consumption, increased weight, dyslipidemia and high HbA1c in men were crucial. Men were in the moderate-risk group according to three risk models. Women were in the medium-risk group according to the Framingham and PROCAM risks and in the low-risk group according to the SCORE. Results estimating the 10-year CVD according to the three risk models were inconsistent. More sensitive CVD risk calculators are needed. Discussion and practice implications: Our results could guide diabetes specialists in identifying gender-specific risks and designing preventive interventions.


Racial and ethnic differences associated with feeding- and activity-related behaviors in infants.
To examine parental reports of feeding and activity behaviors in a cohort of parents of 2-month-olds and how they differ by race/ethnicity. Parents participating in Greenlight, a cluster, randomized trial of obesity prevention at 4 health centers, were queried at enrollment about feeding and activity behaviors thought to increase obesity risk. Unadjusted associations between race/ethnicity and the outcomes of interest were performed by using Pearson χ(2) and Kruskal-Wallis tests. Adjusted analyses were performed by using proportional odds logistic regressions. Eight hundred sixty-three parents (50% Hispanic, 27% black, 18% white; 86% Medicaid) were enrolled. Exclusive formula feeding was more than twice as common (45%) as exclusive breastfeeding (19%); 12% had already introduced solid food; 43% put infants to bed with bottles; 23% propped bottles; 20% always fed when the infant cried; 38% always tried to get children to finish milk; 90% were exposed to television (mean, 346 minutes/day); 50% reported active television watching (mean, 25 minutes/day); and 66% did not meet "tummy time" recommendations. Compared with white parents, black parents were more likely to put children to bed with a bottle (adjusted odds ratio [aOR] = 1.97, P < .004; bottle propping, aOR = 3.1, P < .001), and report more television watching (aOR = 1.6, P = .034). Hispanic parents were more likely than white parents to encourage children to finish feeding (aOR = 1.9, P = .007), bottle propping (aOR = 2.5, P = .009), and report less tummy time (aOR = 0.6, P = .037). Behaviors thought to relate to later obesity were highly prevalent in this large, diverse sample and varied by race/ethnicity, suggesting the importance of early and culturally-adapted interventions.


Early and late mortality after malaria in young children in Papua, Indonesia.
In southern Papua, Indonesia, malaria is highly prevalent in young children and is a significant cause of morbidity and early mortality. The association between malaria and delayed mortality is unknown. Routinely-collected hospital surveillance data from southern Papua, Indonesia, were used to assess the risk of recurrent malaria and mortality within 12 months of an initial presentation with malaria in all children younger than 5 years old attending the local hospital. Analysis was primarily by Kaplan Meier and Cox regression methods. In total 15,716 children presenting with malaria between April 2004 and December 2013 were included in the analysis; 6184 (39.3%) with Plasmodium falciparum, 7499 (47.7%) with P. vivax, 203 (1.3%) with P. malariae, 3 with P. ovale and 1827 (11.6%) with mixed infections. Within 1 year, 48.4% (7620/15,716) of children represented a total of 16,957 times with malaria (range 1 to 11 episodes), with the incidence of malaria being greater in patients initially presenting with P. vivax infection (1334 [95%CI 1307-1361] per 1000 patient years) compared to those with P. falciparum infection (920 [896-944]). In total 266 (1.7%) children died within 1 year of their initial presentation, 129 (48.5%) within 30 days and 137 (51.5%) between 31 and 365 days. There was no significant difference in the mortality risk in patients infected with P. vivax versus P. falciparum either before 30 days (Hazard Ratio (HR) 1.02 [0.69,1.49]) or between 31 and 365 days (HR = 1.30 [0.90,1.88]). Children who died had a greater incidence of malaria, 2280 [95%CI 1946-2671] per 1000 patient years preceding their death, compared to 1141 [95%CI 1124-1158] per 1000 patient years in those surviving. Children under-5 years old with P. vivax malaria, are at significant risk of multiple representations with malaria and of dying within 1 year of their initial presentation. Preventing recurrent malaria must be a public health priority in this vulnerable population.


Approaches used to estimate bioavailability when deriving dietary reference values for iron and zinc in adults.
This review aims to describe approaches used to estimate bioavailability when deriving dietary reference values (DRV) for iron and zinc using the factorial approach. Various values have been applied by different expert bodies to convert absorbed iron or zinc into dietary intakes, and these are summarised in this review. The European Food Safety Authority (EFSA) derived zinc requirements from a trivariate saturation response model describing the relationship between zinc absorption and dietary zinc and phytate. The average requirement for men and women was determined as the intercept of the total absorbed zinc needed to meet physiological requirements, calculated according to body weight, with phytate intake levels of 300, 600, 900 and 1200 mg/d, which are representative of mean/median intakes observed in European populations. For iron, the method employed by EFSA was to use whole body iron losses, determined from radioisotope dilution studies, to calculate the quantity of absorbed iron required to maintain null balance. Absorption from the diet was estimated from a probability model based on measures of iron intake and status and physiological requirements for absorbed iron. Average dietary requirements were derived for men and pre- and post-menopausal women. Taking into consideration the complexity of deriving DRV for iron and zinc, mainly due to the limited knowledge on dietary bioavailability, it appears that EFSA has made maximum use of the most relevant up-to-date data to develop novel and transparent DRV for these nutrients.


The interaction between Toxoplasma gondii and mammalian cells. II. The absence of lysosomal fusion with phagocytic vacuoles containing living parasites.
Electron microscope methods have been used to study delivery of macrophage primary or secondary lysosomal contents to phagocytic vacuoles containing living or dead toxoplasmas. Secondary lysosomes were labeled by culturing the cells in colloidal thorium dioxide (thorotrast) or in ferritin. Acid phosphatase cytochemistry was employed for detection of primary as well as secondary lysosomal constituents. These various lysosomal labels were present in nearly all vacuoles containing toxoplasmas killed with glutaraldehyde, or in vacuoles containing those parasites undergoing degeneration 1 hr after the uptake of living toxoplasmas. In contrast, at times ranging from 1 to 20 hr after infection, no vacuoles containing morphologically normal, apparently viable toxoplasmas were thorotrast or ferritin positive, and only rarely did these vacuoles react for acid phosphatase. In many instances vacuoles containing viable toxoplasmas and no lysosomal markers were situated in the same cell nearby to vacuoles containing degenerating toxoplasmas and lysosomal constituents, thus indicating that the determinants of lysosomal fusion were operating locally in the immediate vicinity of the phagocytic vacuole, and not operating to influence general cell function. Thus, some toxoplasmas are able to prevent the delivery of lysosomal contents, and apparently the phagocytic vacuole provides for these parasites a sheltered microenvironment ideal for their growth. Morphologic evidence indicated that living toxoplasmas altered the phagocytic vacuolar membrane in macrophages, fibroblasts, and HeLa cells. Within minutes after phagocytosis, the vacuole became surrounded by closely apposed strips of endoplasmic reticulum and mitochondria; somewhat later, microvillous protrusions of the membrane into the vacuole were seen. These morphologic features of phagocytic vacuoles containing living toxoplasmas may be of importance in relation to the absence of lysosomal fusion, or they may serve some function in protecting the host cell or in nourishing the parasite.


Glycomics of a novel type-2 N-acetyllactosamine-specific lectin purified from the feather star, Oxycomanthus japonicus (Pelmatozoa: Crinoidea).
A lectin - designated OXYL for the purposes of this study that strongly recognizes complex-type oligosaccharides of serum glycoproteins - was purified from a crinoid, the feather star Oxycomanthus japonicus, the most basal group among extant echinoderms. OXYL was purified through a combination of anion-exchange and affinity chromatography using Q-sepharose and fetuin-sepharose gel, respectively. Lectin was determined to be a 14-kDa polypeptide by sodium dodecyl sulphate-polyacrylamide gel electrophoresis under reducing conditions. However, 14-kDa and 28-kDa bands appeared in the same proportion under non-reducing conditions. Gel permeation chromatography showed a 54-kDa peak, suggesting that lectin consists of four 14-kDa subunits. Divalent cations were not indicated, and stable haemagglutination activity was demonstrated at pH 4-12 and temperatures below 60°C. Surface plasmon resonance analysis of OXYL against fetuin showed k(ass) and k(diss) values of 1.4×10(-6)M(-1)s(-1) and 3.1×10(-3)s(-1), respectively, indicating that it has a strong binding affinity to the glycoprotein as lectin. Frontal affinity chromatography using 25 types of prydylamine-conjugated glycans indicated that OXYL specifically recognizes multi-antennary complex-type oligosaccharides containing type-2 N-acetyllactosamines (Galβ1-4GlcNAc) if α2-3-linked sialic acid is linked at the non-reducing terminal. However, type-1 N-acetyllactosamine (Galβ1-3GlcNAc) chains and α2-6-linked sialic acids were never recognized by OXYL. This profiling study showed that OXYL essentially recognizes β1-4-linkage at C-1 position and free OH group at C-6 position of Gal in addition to the conservation of N-acetyl groups at C-2 position and free OH groups at C-3 position of GlcNAc in N-acetyllactosamine. This is the first report on glycomics on a lectin purified from an echinoderm belonging to the subphylum Pelmatozoa.


1-[11C]-acetate PET imaging in head and neck cancer--a comparison with 18F-FDG-PET: implications for staging and radiotherapy planning.
The aim of this study was to evaluate the feasibility of using 1-[(11)C]-acetate positron emission tomography (ACE-PET) to detect and delineate the gross tumour volume of head and neck cancer before radiotherapy, and to compare the results with those obtained using (18)F-fluoro-2-deoxy-D: -glucose (FDG) PET. Ten patients with histologically verified squamous cell carcinoma were investigated by FDG-PET and dynamic ACE-PET prior to radiotherapy. The two scans were performed on the same day or on consecutive days, except in one patient in whom they were done 5 days apart. Diagnostic CT or MRI was performed in all patients. The image data sets were analysed both visually and semi-quantitatively. All primary tumours and metastases were delineated automatically by using the 50% threshold of maximum radioactivity corrected for background. The mean standardised uptake value (SUV) and the tumour volumes were evaluated and compared. All ten primary tumours were detected by ACE-PET, while nine primaries were detected by FDG-PET and CT and/or MRI. The ACE SUV tended to be lower than the FDG SUV (5.3+/-2.7 vs 9.6+/-7.0, p=0.07). The tumour volumes delineated with ACE were on average 51% larger than the FDG volumes (p<0.05). ACE-PET identified 20/21 lymph node metastases, while only 13/21 lesions were detected by FDG-PET and 16/21 lesions by CT or MRI. ACE-PET appears promising for the staging of head and neck cancer. The biological information provided by both FDG and ACE must be carefully validated before it can be used in clinical routine for radiation treatment planning. More studies are needed to evaluate the differences in volumes and to confirm the clinical potential of both FDG and ACE-PET, especially in radiotherapy.


ScreenPro FH - Screening Project for Familial Hypercholesterolemia in Central, Southern and Eastern Europe: Basic Epidemiology.
Despite great recent progress, familial hypercholesterolemia (FH) is still underestimated, under-diagnosed and thus undertreated worldwide. We have very little information on exact prevalence of patients with FH in the Central, Eastern and Southern Europe (CESE) region. The aim of the study was to describe the epidemiological situation in the CESE region from data available. All local leaders of the ScreenPro FH project were asked to provide local data on (a) expert guess of FH prevalence (b) the medical facilities focused on FH already in place (c) the diagnostic criteria used (d) the number of patients already evidenced in local database and (e) the availability of therapeutic options (especially plasma apheresis). With the guess prevalence of FH around 1 : 500, we estimate the overall population of 588 363 FH heterozygotes in the CESE region. Only 14 108 persons (2.4 %) were depicted in local databases; but the depiction rate varied between 0.1 % and 31.6 %. Only four out of 17 participating countries reported the the LDL apheresis availability. Our data point to the large population of heterozygous FH patients in the CESE region but low diagnostic rate. However structures through the ScreenPro FH project are being created and we can hope that the results will appear soon.Key words: diagnosis - epidemiology - familial hypercholesterolemia - screening.


Neuropathological background of phenotypical variability in frontotemporal dementia.
Frontotemporal lobar degeneration (FTLD) is the umbrella term encompassing a heterogeneous group of pathological disorders. With recent discoveries, the FTLDs have been show to classify nicely into three main groups based on the major protein deposited in the brain: FTLD-tau, FTLD-TDP and FTLD-FUS. These pathological groups, and their specific pathologies, underlie a number of well-defined clinical syndromes, including three frontotemporal dementia (FTD) variants [behavioral variant frontotemporal dementia (bvFTD), progressive non-fluent aphasia, and semantic dementia (SD)], progressive supranuclear palsy syndrome (PSPS) and corticobasal syndrome (CBS). Understanding the neuropathological background of the phenotypic variability in FTD, PSPS and CBS requires large clinicopathological studies. We review current knowledge on the relationship between the FTLD pathologies and clinical syndromes, and pool data from a number of large clinicopathological studies that collectively provide data on 544 cases. Strong relationships were identified as follows: FTD with motor neuron disease and FTLD-TDP; SD and FTLD-TDP; PSPS and FTLD-tau; and CBS and FTLD-tau. However, the relationship between some of these clinical diagnoses and specific pathologies is not so clear cut. In addition, the clinical diagnosis of bvFTD does not have a strong relationship to any FTLD subtype or specific pathology and therefore remains a diagnostic challenge. Some evidence suggests improved clinicopathological association of bvFTD by further refining clinical characteristics. Unlike FTLD-tau and FTLD-TDP, FTLD-FUS has been less well characterized, with only 69 cases reported. However, there appears to be some associations between clinical phenotypes and FTLD-FUS pathologies. Clinical diagnosis is therefore promising in predicting molecular pathology.


Catheter-directed interventions for pulmonary embolism.
Pulmonary embolism (PE), a potentially life-threatening entity, can be treated medically, surgically, and percutaneously. In patients with right ventricular dysfunction (RVD), anticoagulation alone may be insufficient to restore cardiac function. Because of the morbidity and mortality associated with surgical embolectomy, clinical interest in catheter-directed interventions (CDI) has resurged. We describe specific catheter-directed techniques and the evidence supporting percutaneous treatments.


Retrospective screening of routine respiratory samples revealed undetected community transmission and missed intervention opportunities for SARS-CoV-2 in the United Kingdom.
In the early phases of the SARS coronavirus type 2 (SARS-CoV-2) pandemic, testing focused on individuals fitting a strict case definition involving a limited set of symptoms together with an identified epidemiological risk, such as contact with an infected individual or travel to a high-risk area. To assess whether this impaired our ability to detect and control early introductions of the virus into the UK, we PCR-tested archival specimens collected on admission to a large UK teaching hospital who retrospectively were identified as having a clinical presentation compatible with COVID-19. In addition, we screened available archival specimens submitted for respiratory virus diagnosis, and dating back to early January 2020, for the presence of SARS-CoV-2 RNA. Our data provides evidence for widespread community circulation of SARS-CoV-2 in early February 2020 and into March that was undetected at the time due to restrictive case definitions informing testing policy. Genome sequence data showed that many of these early cases were infected with a distinct lineage of the virus. Sequences obtained from the first officially recorded case in Nottinghamshire - a traveller returning from Daegu, South Korea - also clustered with these early UK sequences suggesting acquisition of the virus occurred in the UK and not Daegu. Analysis of a larger sample of sequences obtained in the Nottinghamshire area revealed multiple viral introductions, mainly in late February and through March. These data highlight the importance of timely and extensive community testing to prevent future widespread transmission of the virus.


Acupuncture and related techniques for postoperative pain: a systematic review of randomized controlled trials.
Postoperative pain management remains a significant challenge for all healthcare providers. The objective of this systematic review was to quantitatively evaluate the efficacy of acupuncture and related techniques as adjunct analgesics for acute postoperative pain management. We searched the databases of Medline (1966-2007), CINAHL, The Cochrane Central Register of Controlled Trials (2006), and Scopus for randomized controlled trials (RCTs) using acupuncture for postoperative pain management. We extracted data about postoperative opioid consumption, postoperative pain intensity, and opioid-related side-effects. Combined data were analysed using a random effects model. Fifteen RCTs comparing acupuncture with sham control in the management of acute postoperative pain were included. Weighted mean difference for cumulative opioid analgesic consumption was -3.14 mg (95% confidence interval, CI: -5.15, -1.14), -8.33 mg (95% CI: -11.06, -5.61), and -9.14 mg (95% CI: -16.07, -2.22) at 8, 24, and 72 h, respectively. Postoperative pain intensity (visual analogue scale, 0-100 mm) was also significantly decreased in the acupuncture group at 8 and 72 h compared with the control group. The acupuncture treatment group was associated with a lower incidence of opioid-related side-effects such as nausea (relative risk, RR: 0.67; 95% CI: 0.53, 0.86), dizziness (RR: 0.65; 95% CI: 0.52, 0.81), sedation (RR: 0.78; 95% CI: 0.61, 0.99), pruritus (RR: 0.75; 95% CI: 0.59, 0.96), and urinary retention (RR: 0.29; 95% CI: 0.12, 0.74). Perioperative acupuncture may be a useful adjunct for acute postoperative pain management.


Prospective study of bone mineral density changes in aging men with or at risk for HIV infection.
To investigate rates and predictors of change in bone mineral density (BMD) in a cohort of aging men with or at risk for HIV infection. A prospective cohort study among 230 HIV-infected and 159 HIV-uninfected men aged at least 49 years. Longitudinal analyses of annual change in BMD at the femoral neck, total hip, and lumbar spine. At baseline, 46% of men had normal BMD, 42% had osteopenia, and 12% had osteoporosis. Of those men with normal BMD, 14% progressed to osteopenia and 86% continued to have normal BMD. Of the men initially with osteopenia, 12% progressed to osteoporosis and 83% continued to have osteopenia. Osteopenia incidence per 100 person-years at risk was 2.6 for HIV-uninfected men and 7.2 for HIV-infected men; osteoporosis incidence was 2.2 per 100 person-years at risk among men with osteopenia, regardless of HIV status. In multivariable analysis of annual change in BMD at the femoral neck, we found a significant interaction between heroin use and AIDS diagnosis, such that the greatest bone loss occurred with both AIDS and heroin use (adjusted predicted mean annual bone loss 0.0196 g/cm). Hepatitis C virus seropositivity was also associated with femoral neck bone loss (P = 0.04). The interaction between AIDS and heroin use also was associated with bone loss at the total hip, as was current methadone use (P < 0.01). We found an association of heroin use and AIDS with BMD change, suggesting that heroin users with AIDS may be at particular risk for bone loss.


Surges of muscle sympathetic nerve activity during obstructive apnea are linked to hypoxemia.
Obstructive sleep apnea (OSA) is associated with oscillations of arterial blood pressure (BP) that occur in phase with irregularities of respiration. To explore the role of the sympathetic nervous system in these responses, we studied muscle sympathetic nerve activity (MSNA; peroneal microneurography), an index of vasoconstrictor nerve traffic, and BP during awake regular breathing and during spontaneous apneas in patients with OSA. To determine the role of the arterial chemoreflex, we also examined the effects of 100% O2 (hyperoxia) on MSNA and BP. In awake regularly breathing patients with OSA (n = 12), resting MSNA was markedly higher than in an age-matched control population (n = 15) [41 +/- 23 (SD) vs. 24 +/- 17 bursts/min; P < 0.05] and was unchanged during hyperoxia (n = 9). Apneas during sleep (n = 8) were associated with surges in MSNA followed by transient rises in BP when breathing resumed. In contrast to room air apneas, hyperoxic apneas of similar duration were associated with attenuated MSNA responses (+82 +/- 84% vs. +5 +/- 25% compared with awake baseline; P < 0.05; n = 6), even though O2 did not affect sleep stage and the occurrence of arousal. Thus the BP oscillations that occur with apnea during sleep may in part be mediated by intermittent surges of sympathetic activity resulting in vasoconstriction. Because the MSNA responses to obstructive apnea are blunted during O2 administration, they appear to be linked to intermittent arterial hypoxemia and stimulation of arterial chemoreceptors.


Real-time activity assays of β-lactamases in living bacterial cells: application to the inhibition of antibiotic-resistant E. coli strains.
The emergence of antibiotic resistance caused by β-lactamases, including serine β-lactamases (SβLs) and metallo-β-lactamases (MβLs), is a global public health threat. L1, a B3 subclass MβL, hydrolyzes almost all of known β-lactam antibiotics. We report a simple and straightforward UV-Vis approach for real-time activity assays of β-lactamases inside living bacterial cells, and this method has been exemplified by choosing antibiotics, L1 enzyme, Escherichia coli expressing L1 (L1 E. coli), Escherichia coli expressing extended-spectrum β-lactamases (ESBL-E. coli), clinical bacterial strains, and reported MβL and SβL inhibitors. The cell-based studies demonstrated that cefazolin was hydrolyzed by L1 E. coli and clinical strains, and confirmed the hydrolysis to be inhibited by two known L1 inhibitors EDTA and azolylthioacetamide (ATAA), with an IC50 value of 1.6 and 18.9 μM, respectively. Also, it has been confirmed that the breakdown of cefazolin caused by ESBL-E. coli was inhibited by clavulanic acid, the first SβL inhibitor approved by FDA. The data gained through this approach are closely related to the biological function of the target enzyme in its physiological environment. The UV-Vis method proposed here can be applied to target-based whole-cell screening to search for potent β-lactamase inhibitors, and to assays of reactions in complex biological systems, for instance in medical assays.


Primary deficiency of microsomal triglyceride transfer protein in human abetalipoproteinemia is associated with loss of CD1 function.
Abetalipoproteinemia (ABL) is a rare Mendelian disorder of lipid metabolism due to genetic deficiency in microsomal triglyceride transfer protein (MTP). It is associated with defects in MTP-mediated lipid transfer onto apolipoprotein B (APOB) and impaired secretion of APOB-containing lipoproteins. Recently, MTP was shown to regulate the CD1 family of lipid antigen-presenting molecules, but little is known about immune function in ABL patients. Here, we have shown that ABL is characterized by immune defects affecting presentation of self and microbial lipid antigens by group 1 (CD1a, CD1b, CD1c) and group 2 (CD1d) CD1 molecules. In dendritic cells isolated from ABL patients, MTP deficiency was associated with increased proteasomal degradation of group 1 CD1 molecules. Although CD1d escaped degradation, it was unable to load antigens and exhibited functional defects similar to those affecting the group 1 CD1 molecules. The reduction in CD1 function resulted in impaired activation of CD1-restricted T and invariant natural killer T (iNKT) cells and reduced numbers and phenotypic alterations of iNKT cells consistent with central and peripheral CD1 defects in vivo. These data highlight MTP as a unique regulator of human metabolic and immune pathways and reveal that ABL is not only a disorder of lipid metabolism but also an immune disease involving CD1.


Executive and mnemonic functions in early Huntington's disease.
Eighteen patients with early Huntington's disease were compared with age- and IQ-matched control volunteers on tests of executive and mnemonic function taken from the Cambridge Neuropsychological Test Automated Battery. Tests of pattern and spatial recognition memory, spatial span, spatial working memory, spatial planning and visual discrimination learning/attentional set shifting were employed. These tests have previously been found to be sensitive to the later stages of Huntington's disease. Patients with early Huntington's disease were found to have a wide range of cognitive impairments encompassing both visuospatial memory and executive functions, a pattern distinct from those seen in other basal ganglia disorders. In contrast to patients with more advanced Huntington's disease, early Huntington's disease patients were not impaired at simple reversal learning, but were impaired at performing an extradimensional shift (EDS). The results will be discussed in relation to the hypothesized neuropathological staging of Huntington's disease and to the anatomical connectivity of the striatum.


21-hydroxylase deficiency families with HLA identical affected and unaffected sibs.
During our investigations of polymorphisms at, and in the immediate chromosomal vicinity of, the 21-hydroxylase locus in families with 21-hydroxylase deficiency, three families were found to show marked discordance in clinical features of HLA identical subjects. In one family, there is discordance between a boy with the simple virilising form of 21-hydroxylase deficiency and his two younger sisters, who are both HLA identical to their brother, but who have additional salt wasting features. In the other two families, one subject is severely affected and has very high 17-hydroxyprogesterone levels, but has an HLA identical sib who is asymptomatic and shows only slightly raised 17-hydroxyprogesterone levels. In all cases, HLA identity, as indicated by protein polymorphism studies (HLA-A, B, DR, C4A, C4B, and Bf typing), has been verified at the gene organisation level using 21-hydroxylase and complement C4 DNA probes. An HLA-Bw47 bearing haplotype in one of the latter families has not been transmitted to the affected child and appears to carry a normal 21-OHB allele and two genes which specify C4A allotypes.


Investigation of the nutraceutical potential of monofloral Indian mustard bee pollen.
This study was designed to investigate the nutraceutical potential of monofloral Indian mustard bee pollen (MIMBP). The nutritional value of MIMBP was examined in terms of proteins, fats, carbohydrates, and energy value. Its chemical composition in terms of total polyphenol and flavonoid content was determined. MIMBP was screened for free flavonoid aglycones by developing and validating a high-performance liquid chromatography-photo diode array (HPLC-PDA) method. MIMBP was analyzed for in vitro antioxidant effect in terms of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity. MIMBP was found to be comprised of proteins ((182.2±5.9) g/kg), fats ((137.7±6.8) g/kg) and carbohydrates ((560.6±17.4) g/kg), which result in its high energy value ((17 616.7±78.6) kJ/kg). MIMBP was found to contain polyphenols ((18 286.1±374.0) mg gallic acid equivalent/kg) and flavonoids ((1 223.5±53.1) mg quercetin equivalent/kg). The HPLC-PDA analysis revealed the presence of kaempferol ((65.4±0.5) mg/kg) and quercetin ((51.4±0.4) mg/kg) in MIMBP, which can be used as markers for determining the quality of bee pollen. The MIMBP extract showed DPPH free radical-scavenging activity with a half maximal inhibitory concentration of 54.79 μg/mL. The MIMBP was found to be a rich source of nutrients providing high caloric value, which makes it a candidate for a potential nutraceutical agent. The study also illustrated the high antioxidant content of MIMBP, especially in the principle polyphenols and flavonoids, which suggests its potential role in the prevention of free radical-implicated diseases. The DPPH-scavenging effect of MIMBP further confirmed its antioxidant potential. Additionally, we developed a simple, specific and accurate HPLC-PDA method for the identification and quantification of free flavonoid aglycones. This can be applied in future screenings of the quality of pollen collected by honeybees.


An IL-23R/IL-22 Circuit Regulates Epithelial Serum Amyloid A to Promote Local Effector Th17 Responses.
RORγt(+) Th17 cells are important for mucosal defenses but also contribute to autoimmune disease. They accumulate in the intestine in response to microbiota and produce IL-17 cytokines. Segmented filamentous bacteria (SFB) are Th17-inducing commensals that potentiate autoimmunity in mice. RORγt(+) T cells were induced in mesenteric lymph nodes early after SFB colonization and distributed across different segments of the gastrointestinal tract. However, robust IL-17A production was restricted to the ileum, where SFB makes direct contact with the epithelium and induces serum amyloid A proteins 1 and 2 (SAA1/2), which promote local IL-17A expression in RORγt(+) T cells. We identified an SFB-dependent role of type 3 innate lymphoid cells (ILC3), which secreted IL-22 that induced epithelial SAA production in a Stat3-dependent manner. This highlights the critical role of tissue microenvironment in activating effector functions of committed Th17 cells, which may have important implications for how these cells contribute to inflammatory disease.


Graduated licensing laws and fatal crashes of teenage drivers: a national study.
The objective of the current study was to quantify the effects of the strength of US state graduated driver licensing laws and specific licensing components on the rate of teenage driver fatal crash involvements per 100,000 teenagers during 1996-2007. The strengths of state laws were rated good, fair, marginal, or poor based on a system developed previously by the Insurance Institute for Highway Safety. Analysis was based on quarterly counts of drivers involved in fatal crashes. Associations of overall ratings and individual licensing components with teenage crash rates were evaluated using Poisson regression, with the corresponding fatal crash rate for drivers ages 30-59 controlling for state- or time-dependent influences on crash rates unrelated to graduated licensing laws. Compared with licensing laws rated poor, laws rated good were associated with 30 percent lower fatal crash rates among 15- to 17-year-olds. Laws rated fair yielded fatal crash rates 11 percent lower. The longer the permit age was delayed, or the longer the licensing age was delayed, the lower the estimated fatal crash rates among 15- to 17-year-olds. Stronger nighttime restrictions were associated with larger reductions, and reductions were larger for laws limiting teenage passengers to zero or one than laws allowing two or more teenage passengers or laws without passenger restrictions. After the effects of any related delay in licensure were accounted for, an increase in the minimum learner's permit holding period showed no association with fatal crash rates. An increase in required practice driving hours did not appear to have an independent association with fatal crash rates. Graduated licensing laws that include strong nighttime and passenger restrictions and laws that delay the learner's permit age and licensing age are associated with lower teenage fatal crash rates. States that adopt such laws can expect to achieve substantial reductions in crash deaths.


Epidemiology of malaria in Rohingya refugee camps in Bangladesh within 2017-2020.
Malaria causes significant morbidity and mortality in tropical and sub-tropical regions, particularly in humanitarian emergencies including refugee camps in malaria endemic areas. An epidemiological investigation was conducted on malaria disease distribution and risk factors in the world's largest refugee settlement, the Rohingya refugee camps on the south-eastern border area of Bangladesh, within 2017-2020. From February 2017 to March 2020, 30,460 febrile patients were tested for malaria using light microscopy and rapid diagnostic tests. Most were self-presenting symptomatic patients and a minority were from door-to-door malaria screening. Diagnostic tests were done by trained medical technologists upon the advice of the concerned physicians in the camps. Test positivity rate (%) and annual parasite incidence were calculated and compared using chi-squared (χ 2) test or odds ratios. The overall average annual test positivity rate (TPR) was 0.05%. TPR was highest in people who had travelled to the forest in the previous 2 months, at 13.60%. Cases were clustered among male adults aged 15-60 years. There were no cases among children under five years or pregnant women and no deaths from malaria. This study found very few malaria cases among Rohingya refugees with the majority of cases being imported from hilly forested areas, which were thus assumed to act as the reservoir for transmission.


Postpolio syndrome: a 5 year follow-up.
The purpose of this investigation was to study subjective symptoms, medical and social situation, pulmonary function and physical work capacity during a period of 3-5 years in patients with the postpolio syndrome. We evaluated 68 patients consecutively admitted to our hospital because of postpolio syndrome, and re-evaluated 63 of the same patients 3-5 years later, 43 women and 20 men with mean age 55 +/- 10 (1 SD) years at the second evaluation. The patients answered a questionnaire about their subjective symptoms and medical and social situation, and underwent spirometry as well as symptom-limited exercise stress testing. Most patients experienced increasing symptoms and physical disability related to their polio, while the majority reported that their mental health were unchanged or improved. The lung function was in average moderately reduced of restrictive type, and only minor changes were found during the 3-5 years. A pronounced reduction in peak oxygen uptake was seen at the first evaluation, especially in women. At the second examination, peak oxygen uptake was further decreased, especially in men, more than predicted from increasing age. The patients increased their body mass index significantly during the same period. These results indicate that subjective symptoms and physical disability related to polio increased with increasing age in these patients with the post-polio syndrome, and cardiorespiratory deconditioning and weight gain also became increasing problems in most patients. However, the mental status of the patients remained stable or improved, possibly due to our comprehensive re-rehabilitation and educational programme.


Modeling of electrodes and implantable pulse generator cases for the analysis of implant tip heating under MR imaging.
The authors' purpose is to model the case of an implantable pulse generator (IPG) and the electrode of an active implantable medical device using lumped circuit elements in order to analyze their effect on radio frequency induced tissue heating problem during a magnetic resonance imaging (MRI) examination. In this study, IPG case and electrode are modeled with a voltage source and impedance. Values of these parameters are found using the modified transmission line method (MoTLiM) and the method of moments (MoM) simulations. Once the parameter values of an electrode/IPG case model are determined, they can be connected to any lead, and tip heating can be analyzed. To validate these models, both MoM simulations and MR experiments were used. The induced currents on the leads with the IPG case or electrode connections were solved using the proposed models and the MoTLiM. These results were compared with the MoM simulations. In addition, an electrode was connected to a lead via an inductor. The dissipated power on the electrode was calculated using the MoTLiM by changing the inductance and the results were compared with the specific absorption rate results that were obtained using MoM. Then, MRI experiments were conducted to test the IPG case and the electrode models. To test the IPG case, a bare lead was connected to the case and placed inside a uniform phantom. During a MRI scan, the temperature rise at the lead was measured by changing the lead length. The power at the lead tip for the same scenario was also calculated using the IPG case model and MoTLiM. Then, an electrode was connected to a lead via an inductor and placed inside a uniform phantom. During a MRI scan, the temperature rise at the electrode was measured by changing the inductance and compared with the dissipated power on the electrode resistance. The induced currents on leads with the IPG case or electrode connection were solved for using the combination of the MoTLiM and the proposed lumped circuit models. These results were compared with those from the MoM simulations. The mean square error was less than 9%. During the MRI experiments, when the IPG case was introduced, the resonance lengths were calculated to have an error less than 13%. Also the change in tip temperature rise at resonance lengths was predicted with less than 4% error. For the electrode experiments, the value of the matching impedance was predicted with an error less than 1%. Electrical models for the IPG case and electrode are suggested, and the method is proposed to determine the parameter values. The concept of matching of the electrode to the lead is clarified using the defined electrode impedance and the lead Thevenin impedance. The effect of the IPG case and electrode on tip heating can be predicted using the proposed theory. With these models, understanding the tissue heating due to the implants becomes easier. Also, these models are beneficial for implant safety testers and designers. Using these models, worst case conditions can be determined and the corresponding implant test experiments can be planned.


Rectal sensitivity in diabetes patients with symptoms of gastroparesis.
In a clinical setting, diabetic autonomic complications (cardiac, gastrointestinal, urogenital, etc.) are often handled as separate entities. We investigated rectal sensitivity to heat, mechanical distension, and electrical stimulations in 20 patients with diabetes and symptoms of gastroparesis, to evaluate the extent of visceral neuronal damage. Furthermore, to evaluate the relation between the nervous structures we examined gastric emptying and cardiac autonomic function with the hypothesis being an association between these. We found that 60% of patients had delayed gastric empting. Rectal hyposensitivity was a general finding as they tolerated 67% higher thermal, 42% more mechanical, and 33% higher electrical current intensity compared to healthy controls. In patients, most heart rate variability parameters were reduced; they reported significantly more gastrointestinal symptoms and a reduced quality of life in all SF-36 domains. Shortened RR interval correlated with reduced rectal temperature sensitivity, and gastric retention rate was negatively associated with symptoms of nausea and vomiting. To conclude, in these patients with signs and symptoms of diabetic gastroparesis, rectal sensitivity was reduced, and heart rate variability was impaired. Thus, we suggest regarding diabetic autonomic neuropathy as a diffuse disorder. Symptoms of widespread autonomic dysfunction and sensory disorders should be expected and treated in these patients.


Laser speckle contrast imaging of blood flow in rat retinas using an endoscope.
Laser speckle contrast imaging (LSCI) offers a cost-effective means to image blood flow in vivo. However, it is not commonly used to image rodent retinas because of the challenges associated with imaging through the curved cornea and delivering light through the highly scattering lens. A solution to overcome these problems by using LSCI in conjunction with an endoscope to obtain high spatiotemporal blood flow images is described. Its utility is demonstrated by imaging blood flow changes in rat retinas using hyperoxic, hypercapnic, and visual (flicker) stimulations. Hypercapnia increases blood flow, hyperoxia decreases blood flow, and visual stimulation increases blood flow in the retina relative to basal conditions. The time-to-peak of the LSCI response to visual stimulation is also measured. This approach may prove useful to investigate dysregulation in blood flow-evoked responses in retinal diseases and to evaluate treatment strategies in rodents.


Estimating Indoor PM2.5 and CO Concentrations in Households in Southern Nepal: The Nepal Cookstove Intervention Trials.
High concentrations of household air pollution (HAP) due to biomass fuel usage with unvented, insufficient combustion devices are thought to be an important health risk factor in South Asia population. To better characterize the indoor concentrations of particulate matter (PM2.5) and carbon monoxide (CO), and to understand their impact on health in rural southern Nepal, this study analyzed daily monitoring data collected with DataRAM pDR-1000 and LASCAR CO data logger in 2980 households using traditional biomass cookstove indoor through the Nepal Cookstove Intervention Trial-Phase I between March 2010 and October 2011. Daily average PM2.5 and CO concentrations collected in area near stove were 1,376 (95% CI, 1,331-1,423) μg/m3 and 10.9 (10.5-11.3) parts per million (ppm) among households with traditional cookstoves. The 95th percentile, hours above 100μg/m3 for PM2.5 or 6ppm for CO, and hours above 1000μg/m3 for PM2.5 or 9ppm for CO were also reported. An algorithm was developed to differentiate stove-influenced (SI) periods from non-stove-influenced (non-SI) periods in monitoring data. Average stove-influenced concentrations were 3,469 (3,350-3,588) μg/m3 for PM2.5 and 21.8 (21.1-22.6) ppm for CO. Dry season significantly increased PM2.5 concentration in all metrics; wood was the cleanest fuel for PM2.5 and CO, while adding dung into the fuel increased concentrations of both pollutants. For studies in rural southern Nepal, CO concentration is not a viable surrogate for PM2.5 concentrations based on the low correlation between these measures. In sum, this study filled a gap in knowledge on HAP in rural Nepal using traditional cookstoves and revealed very high concentrations in these households.


Hybrid optical antenna with high directivity gain.
Coupling of a far-field optical mode to electronic states of a quantum absorber or emitter is a crucial process in many applications, including infrared sensors, single molecule spectroscopy, and quantum metrology. In particular, achieving high quantum efficiency for a system with a deep subwavelength quantum absorber/emitter has remained desirable. In this Letter, a hybrid optical antenna based on coupling of a photonic nanojet to a metallo-dielectric antenna is proposed, which allows such efficient coupling. A quantum efficiency of about 50% is predicted for a semiconductor with volume of ~λ³/170. Despite the weak optical absorption coefficient of 2000 cm(-1) in the long infrared wavelength of ~8 μm, very strong far-field coupling has been achieved, as evidenced by an axial directivity gain of 16 dB, which is only 3 dB below of theoretical limit. Unlike the common phased array antenna, this structure does not require coherent sources to achieve a high directivity. The quantum efficiency and directivity gain are more than an order of magnitude higher than existing metallic, dielectric, or metallo-dielectric optical antenna.


Identity development in adolescents with mental problems.
In the revision of the Diagnostic and Statistical Manual (DSM-5), "Identity" is an essential diagnostic criterion for personality disorders (self-related personality functioning) in the alternative approach to the diagnosis of personality disorders in Section III of DSM-5. Integrating a broad range of established identity concepts, AIDA (Assessment of Identity Development in Adolescence) is a new questionnaire to assess pathology-related identity development in healthy and disturbed adolescents aged 12 to 18 years. Aim of the present study is to investigate differences in identity development between adolescents with different psychiatric diagnoses. Participants were 86 adolescent psychiatric in- and outpatients aged 12 to 18 years. The test set includes the questionnaire AIDA and two semi-structured psychiatric interviews (SCID-II, K-DIPS). The patients were assigned to three diagnostic groups (personality disorders, internalizing disorders, externalizing disorders). Differences were analyzed by multivariate analysis of variance MANOVA. In line with our hypotheses, patients with personality disorders showed the highest scores in all AIDA scales with T>70. Patients with externalizing disorders showed scores in an average range compared to population norms, while patients with internalizing disorders lay in between with scores around T=60. The AIDA total score was highly significant between the groups with a remarkable effect size of f= 0.44. Impairment of identity development differs between adolescent patients with different forms of mental disorders. The AIDA questionnaire is able to discriminate between these groups. This may help to improve assessment and treatment of adolescents with severe psychiatric problems.


Mitochondria are intracellular magnesium stores: investigation by simultaneous fluorescent imagings in PC12 cells.
To determine the nature of intracellular Mg2+ stores and Mg2+ release mechanisms in differentiated PC12 cells, Mg2+ and Ca2+ mobilizations were measured simultaneously in living cells with KMG-104, a fluorescent Mg2+ indicator, and fura-2, respectively. Treatment with the mitochondrial uncoupler, carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone (FCCP), increased both the intracellular Mg2+ concentration ([Mg2+]i) and the [Ca2+]i in these cells. Possible candidates as intracellular Mg2+ stores under these conditions include intracellular divalent cation binding sites, endoplasmic reticulum (ER), Mg-ATP and mitochondria. Given that no change in [Mg2+]i was induced by caffeine application, intracellular IP3 or Ca2+ liberated by photolysis, it appears that no Mg2+ release mechanism thus exists that is mediated via the action of Ca2+ on membrane-bound receptors in the ER or via the offloading of Mg2+ from binding sites as a result of the increased [Ca2+]i. FCCP treatment for 2 min did not alter the intracellular ATP content, indicating that Mg2+ was not released from Mg-ATP, at least in the first 2 min following exposure to FCCP. FCCP-induced [Mg2+]i increase was observed at mitochondria localized area, and vice versa. These results suggest that the mitochondria serve as the intracellular Mg2+ store in PC12 cell. Simultaneous measurements of [Ca2+]i and mitochondrial membrane potential, and also of [Ca2+]i and [Mg2+]i, revealed that the initial rise in [Mg2+]i followed that of mitochondrial depolarization for several seconds. These findings show that the source of Mg2+ in the FCCP-induced [Mg2+]i increase in PC12 cells is mitochondria, and that mitochondrial depolarization triggers the Mg2+ release.


Clinical consequences of circulating CD28-negative T cells for solid organ transplantation.
CD28 is an important costimulatory molecule expressed on T cells, interacting with CD80 and CD86 on antigen-presenting cells. All naïve T cells express CD28, but memory T cells may become CD28 negative (CD28neg) as a result of repetitive cell divisions, the influence of TNF-alpha, and infection with cytomegalovirus. This results in accumulation of CD28neg T cells, which may constitute >50% of the total circulating T-cell population in the elderly. The frequency of CD28neg T cells is associated with diseases such as cancer, autoimmunity, and atherosclerosis in which they may act as either effector or suppressor cells. This functional heterogeneity probably underlies the finding that the CD8posCD28neg T-cell population harbors effector cells mediating acute allograft rejection, but also suppressor cells. The CD4posCD28neg T cells are predominantly considered to be a nonclassical risk factor for atherosclerotic disease, and their contribution to alloreactivity is less clear. CD28neg T cells depend on IL-15 for their proliferation, and they have increased the expression of specific costimulatory molecules, such as NK receptors, TNF-alpha receptor family members, and the adhesion/costimulatory molecule CD2. Targeting these costimulatory pathways presents potential therapeutic options to block allograft rejection mediated by CD28neg T cells which are resistant to belatacept.


Female advantage in verbal memory: Evidence of sex-specific cognitive reserve.
We investigated sex differences in verbal memory across different levels of neural dysfunction, measured by temporal lobe glucose metabolic rates (TLGluMR). Three hundred ninety controls and 672 participants with amnestic mild cognitive impairment (aMCI) and 254 with Alzheimer disease (AD) dementia from the Alzheimer's Disease Neuroimaging Initiative completed the Rey Auditory Verbal Learning Test (RAVLT) and [18F]-fluorodeoxyglucose-PET. Cross-sectional analyses were conducted using linear regression to examine the sex by TLGluMR interaction on RAVLT performance in the overall sample and within diagnostic groups adjusting for age, education, and APOE ε4 genotype. Across groups, female sex and higher TLGluMR and their interaction were associated with better verbal memory (p values ≤ 0.005). The female advantage in verbal memory varied by TLGluMR such that the advantage was greatest among individuals with moderate to high TLGluMR and minimal or absent among individuals with lower TLGluMR. Diagnosis-stratified analyses revealed that this interaction was driven by the aMCI group (p values = 0.009). The interaction was not significant in control and AD dementia groups. Women show better verbal memory than men in aMCI despite similar levels of brain hypometabolism. The lifelong advantage that females show over males in verbal memory might represent a form of cognitive reserve that delays verbal memory decline until more advanced pathology, as indexed by TLGluMR. This issue is clinically important because verbal memory scores are used in diagnosing aMCI and AD dementia.


Iterative image-domain decomposition for dual-energy CT.
Dual energy CT (DECT) imaging plays an important role in advanced imaging applications due to its capability of material decomposition. Direct decomposition via matrix inversion suffers from significant degradation of image signal-to-noise ratios, which reduces clinical values of DECT. Existing denoising algorithms achieve suboptimal performance since they suppress image noise either before or after the decomposition and do not fully explore the noise statistical properties of the decomposition process. In this work, the authors propose an iterative image-domain decomposition method for noise suppression in DECT, using the full variance-covariance matrix of the decomposed images. The proposed algorithm is formulated in the form of least-square estimation with smoothness regularization. Based on the design principles of a best linear unbiased estimator, the authors include the inverse of the estimated variance-covariance matrix of the decomposed images as the penalty weight in the least-square term. The regularization term enforces the image smoothness by calculating the square sum of neighboring pixel value differences. To retain the boundary sharpness of the decomposed images, the authors detect the edges in the CT images before decomposition. These edge pixels have small weights in the calculation of the regularization term. Distinct from the existing denoising algorithms applied on the images before or after decomposition, the method has an iterative process for noise suppression, with decomposition performed in each iteration. The authors implement the proposed algorithm using a standard conjugate gradient algorithm. The method performance is evaluated using an evaluation phantom (Catphan©600) and an anthropomorphic head phantom. The results are compared with those generated using direct matrix inversion with no noise suppression, a denoising method applied on the decomposed images, and an existing algorithm with similar formulation as the proposed method but with an edge-preserving regularization term. On the Catphan phantom, the method maintains the same spatial resolution on the decomposed images as that of the CT images before decomposition (8 pairs/cm) while significantly reducing their noise standard deviation. Compared to that obtained by the direct matrix inversion, the noise standard deviation in the images decomposed by the proposed algorithm is reduced by over 98%. Without considering the noise correlation properties in the formulation, the denoising scheme degrades the spatial resolution to 6 pairs/cm for the same level of noise suppression. Compared to the edge-preserving algorithm, the method achieves better low-contrast detectability. A quantitative study is performed on the contrast-rod slice of Catphan phantom. The proposed method achieves lower electron density measurement error as compared to that by the direct matrix inversion, and significantly reduces the error variation by over 97%. On the head phantom, the method reduces the noise standard deviation of decomposed images by over 97% without blurring the sinus structures. The authors propose an iterative image-domain decomposition method for DECT. The method combines noise suppression and material decomposition into an iterative process and achieves both goals simultaneously. By exploring the full variance-covariance properties of the decomposed images and utilizing the edge predetection, the proposed algorithm shows superior performance on noise suppression with high image spatial resolution and low-contrast detectability.


Anti-diabetic activity of a polyphenol-rich extract from Phellinus igniarius in KK-Ay mice with spontaneous type 2 diabetes mellitus.
The present study investigated the anti-diabetic activity and potential mechanisms of the polyphenol rich extract from Phellinus igniarius (PI-PRE) in vitro and in vivo. Four main phenolic compounds of PI-PRE were purified and identified as 7,8-dihydroxycoumarin, 3,4-dihydroxybenzalacetone, 7,3'-dihydroxy-5'-methoxyisoflavone and inoscavin C by the off-line semipreparative liquid chromatography-nuclear magnetic resonance protocol. In vitro, PI-PRE stimulated GLUT4 translocation by 2.34-fold and increased glucose uptake by 1.73-fold in L6 cells. However, the selective AMP-activated protein kinase (AMPK) inhibitor, compound C, completely reversed the PI-PRE-induced GLUT4 translocation. In vivo, KK-Ay mice treated with PI-PRE for four weeks had lower fasting blood glucose levels, as well as other blood-lipid indexes, compared with the vehicle control group. Mechanistic studies showed that the expressions of p-AMPKα and GLUT4 were significantly increased by treatment with PI-PRE in L6 cells. In KK-Ay mice, the expression of p-AMPKα was enhanced in the liver and skeletal muscle, and the expression of GLUT4 was increased in skeletal muscle. These findings suggest that PI-PRE possesses potential anti-diabetic effects including improving glucose tolerance, reducing hyperglycemia, and normalizing insulin levels. These effects are partly due to the activation of GLUT4 translocation via the modulation of the AMPK pathway.