Extraction of gastric slow waves from electrogastrograms: combining independent component analysis and adaptive signal enhancement.
The electrogastrogram (EGG), a cutaneous measurement of gastric electrical activity, is a mixture of gastric slow waves and noise. To detect the propagation of gastric slow waves, it is desired to obtain gastric slow waves in each of multichannel EGGs. Recently, independent component analysis (ICA) has shown its efficiency in separating the gastric slow wave from noisy multichannel EGGs. However, this method is not able to recover gastric slow waves in each of the multichannel EGGs. In this paper, a two-stage combined method was proposed for extracting gastric slow waves. First, ICA was performed to separate the gastric slow wave component from noisy multichannel EGGs. Second, adaptive signal enhancement with a reference input derived by the ICA in the first stage was employed to extract gastric slow waves in each channel. Quantitative analysis showed that, with the proposed method, the maximum root-mean-square error between the estimated time lag and its theoretical value in the simulations was only 0.65. The results from real EGG data demonstrated that the combined method was able to extract gastric slow waves from individual channels of EGGs which are important to identify the slow wave propagation. Therefore, the proposed method can be used to detect propagation of gastric slow waves from multichannel EGGs.


Assessing "best evidence": issues in grading the quality of studies for systematic reviews.
Evidence-based medicine, clinical practice guidelines, quality and value of health services, and science-based decision making are becoming mainstays of the health care sector. As part of the evidence-based movement, systematic reviews of the literature on clinical questions are becoming increasingly common. Part of the structured approach to evaluating the literature involves assessing the quality of individual studies included in systematic reviews. To clarify issues in this area, in 1998 the Agency for Health Care Policy and Research commissioned a small project to determine how its 12 Evidence-based Practice Centers were carrying out this part of their systematic reviews (called evidence reports). The number of potential checklists, scales, and similar tools for grading the methodology or the clinical relevance of individual reports is large; the reliability, the validity, the feasibility, and the utility of these tools are either unmeasured or quite variable. Numerous methodologic questions await definitive research and answers, but in the meantime teams developing authoritative systematic reviews can take certain steps to ensure that their approaches to grading the quality of articles meet applicable scientific standards. Clinicians, program administrators, and health policymakers can then be confident in the overall strength of the evidence and study conclusions.


Neuroendocrine Liver Metastasis: Prognostic Implications of Primary Tumor Site on Patients Undergoing Curative Intent Liver Surgery.
Neuroendocrine tumors typically arise from pancreatic (PNET) vs. gastrointestinal or thoracic origins (non-PNET). The impact of primary tumor site on long-term prognosis following resection of neuroendocrine liver metastasis (NELM) remains poorly defined. The objective of the current study was to define the association of primary tumor location on prognosis of patients undergoing curative intent liver resection for NELM. Between 1990 and 2014, 421 patients who underwent resection of NELM were identified from a multi-institutional database. Clinicopathological characteristics, operative details, and outcomes were stratified and analyzed by location of the primary tumor (PNET vs. non-PNET). A propensity score-matched analysis was utilized to assess the impact of primary tumor location on long-term survival. Among the 421 patients, 197 (46.8%) patients had NELM from a PNET primary while 224 (53.2%) had a non-PNET primary (small bowel, n = 145; rectal, n = 10; bronchial, n = 22; other, n = 47). There were no differences in tumor burden and tumor site, while presence of extrahepatic disease was more common among patients with non-PNET NELM (extrahepatic disease, PNET NELM, n = 11 27.5% vs. non-PNET NELM, n = 29 72.5%; p = 0.010). Patients with PNET NELM were more likely to have non-functional disease compared with patients who had non-PNET NELM (non-functional, PNET NELM, n = 117 54.9% vs. non-PNET NELM, n = 96 45.1%; p = 0.011). On the final pathological specimen of the resected NELM, patients with PNET NELM were more likely to have a moderately differentiated tumor (59.3%), while patients with non-PNET NELM were more likely to have a poorly differentiated tumor (67.8%) (p = 0.005). Patients with PNET NELM had a worse 5-year DFS and 5-year OS compared with patients who had non-PNET NELM (DFS, PNET 36.2% vs. non-PNET 55.2%; p = 0.001 and OS, PNET 79.5% vs. non-PNET 83.4%; p = 0.008). After propensity score matching, both 5-year DFS and 5-year OS of the PNET and non-PNET groups were comparable (DFS, PNET 46.2% vs. non-PNET 55.9%; p = 0.22 and OS, PNET 81.5% vs. non-PNET 84.3%; p = 0.19). PNET patients more often present with non-functional NELM and moderately differentiated tumors. On propensity-matched analysis, factors such as extrahepatic disease and tumor grade, but not primary tumor location, were associated with prognosis of patients undergoing curative intent liver surgery for NELM.


In vivo cytosolic H2O2 changes and Ca2+ homeostasis in mouse skeletal muscle.
Hydrogen peroxide (H2O2) and calcium ions (Ca2+) are functional regulators of skeletal muscle contraction and metabolism. Although H2O2 is one of the activators of the type-1 ryanodine receptor (RyR1) in the Ca2+ release channel, the interdependence between H2O2 and Ca2+ dynamics remains unclear. This study tested the following hypotheses using an in vivo model of mouse tibialis anterior (TA) skeletal muscle. 1) Under resting conditions, elevated cytosolic H2O2 concentration ([H2O2]cyto) leads to a concentration-dependent increase in cytosolic Ca2+ concentration ([Ca2+]cyto) through its effect on RyR1; and 2) in hypoxia (cardiac arrest) and muscle contractions (electrical stimulation), increased [H2O2]cyto induces Ca2+ accumulation. Cytosolic H2O2 (HyPer7) and Ca2+ (Fura-2) dynamics were resolved by TA bioimaging in young C57BL/6J male mice under four conditions: 1) elevated exogenous H2O2; 2) cardiac arrest; 3) twitch (1 Hz, 60 s) contractions; and 4) tetanic (30 s) contractions. Exogenous H2O2 (0.1-100 mM) induced a concentration-dependent increase in [H2O2]cyto (+55% at 0.1 mM; +280% at 100 mM) and an increase in [Ca2+]cyto (+3% at 1.0 mM; +8% at 10 mM). This increase in [Ca2+]cyto was inhibited by pharmacological inhibition of RyR1 by dantrolene. Cardiac arrest-induced hypoxia increased [H2O2]cyto (+33%) and [Ca2+]cyto (+20%) 50 min postcardiac arrest. Compared with the exogenous 1.0 mM H2O2 condition, [H2O2]cyto after tetanic muscle contractions rose less than one-tenth as much, whereas [Ca2+]cyto was 4.7-fold higher. In conclusion, substantial increases in [H2O2]cyto levels evoke only modest Ca2+ accumulation via their effect on the sarcoplasmic reticulum RyR1. On the other hand, contrary to hypoxia secondary to cardiac arrest, increases in [H2O2]cyto from muscle contractions are small, indicating that H2O2 generation is unlikely to be a primary factor driving the significant Ca2+ accumulation after, especially tetanic, muscle contractions.NEW & NOTEWORTHY We developed an in vivo mouse myocyte H2O2 imaging model during exogenous H2O2 loading, ischemic hypoxia induced by cardiac arrest, and muscle contractions. In this study, the interrelationship between cytosolic H2O2 levels and Ca2+ homeostasis during muscle contraction and hypoxic conditions was revealed. These results contribute to the elucidation of the mechanisms of muscle fatigue and exercise adaptation.


Prevalence and risk factors associated with sexually transmitted diseases (STDs) in Sikkim.
The population of Sikkim is a unique blend of multi-tribal and metropolitan culture. However, till date, no data regarding prevalence of sexually transmitted diseases (henceforth abbreviated as STDs) among this population is available and hence requires attention. Hence the objective is to determine the prevalence of STDs in Sikkim and to describe associated risk factors. A cross-sectional study involving 'Questionnaire-based anonymous feedback system' was followed to collect data from 2,000 individuals across the society. The four most common STDs, gonorrhea, syphilis, chlamydia and HIV, were considered for the study. Total 69 (3.6%) cases of STDs were found in 1,918 individuals was affected by at least one of the STDs, out of which 43 were males and 26 were females. Cases of gonorrhea, syphilis, chlamydiasis and HIV were 25, 22, 4 and 18 respectively. Out of total 69 cases of STDs, 20 individuals were also suffering from some kind of hepatitis. Addictions like alcoholism, smoking and drugs were also found in significant number, with 1,019 (>50%) individuals with at least one of these addictions. Relative risk analysis indicates that gender-wise females are more vulnerable to STDs than males. The number of partners, addictions, especially alcohol and drug abuse, also contribute to STD cases. STDs act as a significant risk factor in transmitting some of the types of hepatitis. In such cases, females are more vulnerable than males. The results suggest that new community health programs are essential for both, HIV and non-HIV STDs in Sikkim.


Engineered protein disaggregases mitigate toxicity of aberrant prion-like fusion proteins underlying sarcoma.
FUS and EWSR1 are RNA-binding proteins with prion-like domains (PrLDs) that aggregate in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). The FUS and EWSR1 genes are also prone to chromosomal translocation events, which result in aberrant fusions between portions of the PrLDs of FUS and EWSR1 and the transcription factors CHOP and FLI. The resulting fusion proteins, FUS-CHOP and EWS-FLI, drive aberrant transcriptional programs that underpin liposarcoma and Ewing's sarcoma, respectively. The translocated PrLDs alter the expression profiles of these proteins and promote their phase separation and aggregation. Here, we report the development of yeast models of FUS-CHOP and EWS-FLI toxicity and aggregation. These models recapitulated several salient features of sarcoma patient cells harboring the FUS-CHOP and EWS-FLI translocations. To reverse FUS and EWSR1 aggregation, we have explored Hsp104, a hexameric AAA+ protein disaggregase from yeast. Previously, we engineered potentiated Hsp104 variants to suppress the proteotoxicity, aggregation, and mislocalization of FUS and other proteins that aggregate in ALS/FTD and Parkinson's disease. Potentiated Hsp104 variants that robustly suppressed FUS toxicity and aggregation also suppressed the toxicity and aggregation of FUS-CHOP and EWS-FLI. We suggest that these new yeast models are powerful platforms for screening for modulators of FUS-CHOP and EWS-FLI phase separation. Moreover, Hsp104 variants might be employed to combat the toxicity and phase separation of aberrant fusion proteins involved in sarcoma.


Closed-Form Power Normalization Methods for a Satellite MIMO System.
The paper proposes a new set of normalization techniques for precoding/beamforming matrices applicable to broadband multiuser multiple-input multiple-output (MIMO) satellite systems. The proposed techniques adapt known normalization methods to account for the signal attenuation experienced by users due to the degradation of antenna gain and free space losses towards the edge of the coverage. We use, as an example, an array-fed reflector (AFR) antenna onboard a satellite in geosynchronous orbit (GEO), which provides a favorable trade-off between high-directivity, reconfigurability, and the requirement for digital processing, but suffers from high scan losses away from broadside due to optical aberrations when considered for global coverage applications. Three different precoding/beamforming techniques are employed, namely zero forcing (ZF), minimum mean squared error (MMSE), and matched filtering (MF). Low-complexity power normalization techniques digitally applied after the beamformer are introduced that, in the absence of any atmospheric effects, lead to iso-flux-like characteristics whilst satisfying the power constraint per feed. In comparison with other methods reported in the literature, mainly based on iterative algorithms, the proposed techniques consist in closed-form expressions to provide uniform signal-to-noise ratio (SNR) and signal-to-noise plus interference ratio (SNIR) across the users without significant impact on the payload sum rate. Numerical results are presented to comparatively demonstrate the achieved performance in terms of total capacity and distribution of SNR and SNIR at various noise and interference scenarios.


Neuronal nitric oxide synthase negatively regulates xanthine oxidoreductase inhibition of cardiac excitation-contraction coupling.
Although interactions between superoxide (O(2)(.-)) and nitric oxide underlie many physiologic and pathophysiologic processes, regulation of this crosstalk at the enzymatic level is poorly understood. Here, we demonstrate that xanthine oxidoreductase (XOR), a prototypic superoxide O(2)(.-) -producing enzyme, and neuronal nitric oxide synthase (NOS1) coimmunoprecipitate and colocalize in the sarcoplasmic reticulum of cardiac myocytes. Deficiency of NOS1 (but not endothelial NOS, NOS3) leads to profound increases in XOR-mediated O(2)(.-) production, which in turn depresses myocardial excitation-contraction coupling in a manner reversible by XOR inhibition with allopurinol. These data demonstrate a unique interaction between a nitric oxide and an O(2)(.-) -generating enzyme that accounts for crosstalk between these signaling pathways; these findings demonstrate a direct antioxidant mechanism for NOS1 and have pathophysiologic implications for the growing number of disease states in which increased XOR activity plays a role.


Annular vesicular lupus erythematosus.
Disseminated cutaneous lupus erythematosus is a recently defined variant of lupus erythematosus. It presents as migratory areas of annular erythema, but lacks the atrophy, scarring, and follicular plugging associated with discoid lupus erythematosus. The clinical manifestations of this disease fall somewhere in between those associated with classic discoid lupus erythematosus and those associated with systemic lupus erythematosus. A case of disseminated cutaneous lupus erythematosus occurring in a 91-year-old woman is presented. This patient developed areas of subepidermal cleavage resulting in clinically apparent flat vesiculation with subsequent crusting and erosion. Lupus erythematosus must be considered in the differential diagnosis of migratory annular erythema with or without vesiculation.


[Evaluation of diagnostic criteria of syphilitic infection by means of gas chromatography and mass-spectrometry].
In order to estimate possibility of chromatographic criteria application in diagnostics of syphilitic infection, the authors researched in three directions: evaluation of dysmetabolism of carbohydrate, lipid and amino acid components in patients' serum; search of signal compounds that serve as microbe "cooperative sensitivity" activators (lactones, quinolones, furan boron ethers) with evaluation of the risk of luetic infection generalization; indication of organ lesion markers (the brain, liver, bone structures). The factor analysis performed by the researchers allowed to determine priorities of clinical, laboratory and chromatographic data according to their value. The diagnostic value of various diagnostic chromatographic criteria was estimated as follows: the diagnostic sensitivity was 79.6% to 97.2%, the diagnostic specificity was 50.0% to 92.7%, the positive diagnostic predictability was 61.8% to 94.7% and the negative diagnostic predictability was 60.9% to 95.1%.


Prospective assessment of antidonor cellular alloreactivity is a tool for guidance of immunosuppression in kidney transplantation.
Current characterization of the immune risk in renal transplant patients is only focused on the assessment of preformed circulating alloantibodies; however, alloreactive memory T cells are key players in mediating allograft rejection. Immune monitoring of antidonor alloreactive memory/effector T cells using an IFN-γ Elispot has been shown to distinguish patients at risk for immune-mediated graft dysfunction, suggesting a potential tool for immunosuppression individualization. In this nonrandomized study, we prospectively assessed donor and nondonor T-cell alloreactivity in 60 highly alloreactive patients receiving calcineurin inhibitor-based immunosuppression and in non-T-cell alloreactive transplant recipients treated with a calcineurin inhibitor-free regimen. The impact was evaluated using 1-year allograft outcome. We found a strong association between ongoing antidonor T-cell alloreactivity and histological lesions of acute T cell-mediated rejection in 6-month protocol biopsies, distinguishing those patients with better 1-year graft function, regardless of immunosuppression regimen. Interestingly, evidence for enhanced immune regulation, driven by circulating Foxp3-demethylated regulatory T cells, was only observed among patients achieving antidonor T-cell hyporesponsiveness. Thus, prospective evaluation of donor-specific T-cell sensitization may add crucial information on the alloimmune state of transplanted patients to be used in daily clinical practice.


Isopropylthiol emission by bloom-forming Microcystis: Biochemistry, ecophysiology and semiochemistry of a volatile organosulfur compound.
Microcystis species not only produce toxic cyanobacterial blooms, but can be a significant source of taste and odour. Previous studies have associated foul-smelling volatile organic sulfur compounds (VOSCs) with Microcystis blooms, but have largely attributed these compounds to bacterial bloom decomposition. However, earlier reports of the production of isopropylthio compounds by several Microcystis strains suggests that these cyanobacteria may themselves be a source of these VOSCs. Sulphur compounds have been shown to play important semiochemical roles in algal cell protection and grazer interactions in marine systems, but little is known about the production and chemical ecology of freshwater cyanobacterial VOSCs. To address this knowledge gap, we undertook the first detailed investigation of the biochemistry, ecophysiology and semiochemistry of these compounds and their production by Microcystis, and tested the hypothesis that they act as multifunctional semiochemicals in processes related to cell protection and grazer defence. Using short-term incubations and an adapted headspace-GC-MS technique, we investigated VOSC production by axenic and non-axenic strains, and verified that isopropylthio compounds are in fact produced by these cyanobacteria, identifying 5 isopropyl moiety-containing VOSCs (isopropylthiol (ISH), isopropylmethyl sulfide, isopropyl methyl disulfide, diisopropyl disulfide (ISSI) and diisopropyl trisulfide) as well as methanethiol in three strains. Further studies with the axenic strain Microcystis PCC 7806 using different light regimes, metabolic inhibitors (sodium azide, DCMU), the antioxidant enzyme catalase and stable labelled precursors (hydrogencarbonate, acetates and sulfate) demonstrated that ISH is a true exo-metabolite, synthesized via the acetate pathway. It is actively produced and continuously excreted by the cyanobacteria during growth, with minimal internal storage or post-lysis catalytic generation. The molar ratios of the redox pair ISH/ISSI are not directly involved in the photosynthetic and respiratory electron transport chains, but dependant on the redox state of the cell - likely mediated by reactive oxygen species (ROS), as shown by a marked effect of catalase. These results, along with toxicological and behavioural assays using the two aquatic invertebrates Thamnocephalus platyurus and Daphnia magna indicate that ISH plays multiple important physiological and ecological roles. It acts as an effective antioxidant against high ROS levels, as often experienced in surface blooms, it elicits avoidance-related behavioural responses in grazer communities and at high levels, it can be toxic to some invertebrates.


Muscle-tendon unit length changes differ between young and adult sprinters in the first stance phase of sprint running.
The aim of this study was to compare young and adult sprinters on several biomechanical parameters that were previously highlighted as performance-related and to determine the behaviour of several muscle-tendon units (MTU) in the first stance phase following a block start in sprint running. The ground reaction force (GRF) and kinematic data were collected from 16 adult and 21 young well-trained sprinters. No difference between the groups was found in some of the previously highlighted performance-related parameters (ankle joint stiffness, the range of dorsiflexion and plantar flexor moment). Interestingly, the young sprinters showed a greater maximal and mean ratio of horizontal to total GRF, which was mainly attributed to a greater horizontal GRF relative to body mass and resulted in a greater change in horizontal centre of mass (COM) velocity during the stance phase in the young compared with the adult sprinters. Results from the MTU length analyses showed that adult sprinters had more MTU shortening and higher maximal MTU shortening velocities in all plantar flexors and the rectus femoris. Although previously highlighted performance-related parameters could not explain the greater 100 m sprinting times in the adult sprinters, differences were found in the behaviour of the MTU of the plantar flexors and rectus femoris during the first stance phase. The pattern of length changes in these MTUs provides ideal conditions for the use of elastic energy storage and release for power enhancement.


Case Report of a Massive Retropharyngeal Goiter Resulting in Laryngeal Compression.
This is a rare presentation of a morbidly obese male with a massive retropharyngeal goiter causing laryngeal compressive symptoms and unique airway management challenges. Flexible laryngoscopy revealed a retropharyngeal mass circumferentially compressing the oropharynx and supraglottis and preventing the visualization of the glottis. Awake tracheostomy was performed before total thyroidectomy. Airway compression from a goiter typically results from substernal extension, which usually does not cause difficulty with endotracheal intubation. Extensive retropharyngeal extension can cause supraglottic obstruction of the airway. In these cases, optimal management consists of a closely coordinated approach between anesthesia and surgery teams to establish a surgical airway.


The presence of neutrophils causes RANKL expression in periodontal tissue, giving rise to osteoclast formation.
Increased neutrophil infiltration and osteoclast formation are key characteristics of periodontitis. The effect of these neutrophils on osteoclast formation in periodontitis remains unclear. Therefore, we investigated the effects of neutrophils on osteoclast formation in a neutrophil-deficient mouse model of periodontitis. Anti-Ly6G antibody (Ab) was used for neutrophil depletion in two mouse models: periodontitis and air pouch. In the periodontitis experiments, mice were divided into PBS-administered control (C), control Ab-administered periodontitis (P), and anti-Ly6G Ab-administered periodontitis (P + Ly6G) groups. Periodontitis was induced by ligature of mandibular first molars. In the air pouch experiments, mice were divided into PBS-administered (C), LPS and control Ab-administered (LPS), and LPS and anti-Ly6G Ab-administered (LPS + Ly6G) groups. Neutrophil migration into air pouches was induced by LPS injection. Flow cytometry was used to examine CD11b+ Ly6G+ neutrophils in the blood of periodontitis mice and CD11b+ Ly6G+ RANKL+ neutrophils in exudates of air pouch mice. In periodontal tissue, Ly6G+ neutrophil and RANKL+ cell numbers in periodontal ligament and alveolar bone areas were estimated using immunohistochemistry, osteoclast numbers were measured using TRAP assay, and alveolar bone loss was determined by H&E staining. In blood, CD11b+ Ly6G+ neutrophils were found in greater percentage in the P group than in the C group on days 3 and 7. However, the percentage of neutrophils was lower in the P + Ly6G group than in the C and P groups. In periodontal tissue, the numbers of Ly6G+ neutrophils and RANKL+ cells were lower in the P + Ly6G group than in the P group on day 3. Ly6G+ neutrophil numbers decreased more in the P + Ly6G group than in the P group on day 7, but RANKL+ cell numbers did not decrease in the P + Ly6G group. In exudates, the number of CD11b+ Ly6G+ RANKL+ neutrophils was greater in the LPS group than in the C and LPS + Ly6G groups. On days 3 and 7, the numbers of osteoclasts and alveolar bone loss were greater in periodontal tissue in the P and P + Ly6G groups than in the C group. Interestingly, there were fewer osteoclasts in the P + Ly6G group than in the P group on day 3. Neutrophil deficiency caused a reduction in numbers of both RANKL+ cells and osteoclasts in periodontitis-induced tissues only on day 3. Furthermore, in the LPS-injected air pouch model, neutrophil deficiency reduced the influx of RANKL+ neutrophils. These findings suggest that the presence of neutrophils induces RANKL expression and could induce osteoclast formation in the early stages of periodontitis.


Mandibular fractures in children. A retrospective study.
This study reviews the treatment of mandibular fractures in children for the period 1986-1995. The sample included 107 children under 15 years of age, which were studied by age, sex, cause and type of fracture, method of treatment and complications. The children were divided into four groups according to the development of the dentition: 0-5 years, 6-9 years, 10-12 years and 13-15 years. Boys to girls ratio was 3.46:1 and the highest incidence was observed in boys over 13 years of age. Associated injuries were more common in younger children (56%). Most of the fractures were the results of traffic accidents (28.97%), games and sports (25.23%), beating (19.63%) or falls (15.88%). Unilateral fractures were more common (69.16%) than bilateral (30.84%). Intermaxillar fixation for 3 weeks was the method of treatment used in 84.12% of the patients. Complications appeared to be rare.


Functional ecology and evolution of hominoid molar enamel thickness: Pan troglodytes schweinfurthii and Pongo pygmaeus wurmbii.
The divergent molar characteristics of Pan troglodytes and Pongo pygmaeus provide an instructive paradigm for examining the adaptive form-function relationship between molar enamel thickness and food hardness. Although both species exhibit a categorical preference for ripe fruit over other food objects, the thick enamel and crenulated occlusal surface of Pongo molar teeth predict a diet that is more resistant to deformation (hard) and fracture (tough) than the diet of Pan. We confirm these predictions with behavioral observations of Pan troglodytes schweinfurthii and Pongo pygmaeus wurmbii in the wild and describe the mechanical properties of foods utilized during periods when preferred foods are scarce. Such fallback foods may have exerted a selective pressure on tooth evolution, particularly molar enamel thinness, which is interpreted as a functional adaptation to seasonal folivory and a derived character trait within the hominoid clade. The thick enamel and crenulated occlusal surface of Pongo molars is interpreted as a functional adaptation to the routine consumption of relatively tough and hard foods. We discuss the implications of these interpretations for inferring the diet of hominin species, which possessed varying degrees of thick molar enamel. These data, which are among the first reported for hominoid primates, fill an important empirical void for evaluating the mechanical plausibility of putative hominin food objects.


Immunogenicity, Safety, and Efficacy of a Standalone Universal Influenza Vaccine, FLU-v, in Healthy Adults: A Randomized Clinical Trial.
FLU-v is a broad-spectrum influenza vaccine that induces antibodies and cell-mediated immunity. To compare the safety, immunogenicity, and exploratory efficacy of different formulations and dosing regimens of FLU-v versus placebo. Randomized, double-blind, placebo-controlled, single-center phase 2b clinical trial. (ClinicalTrials.gov: NCT02962908; EudraCT: 2015-001932-38). The Netherlands. 175 healthy adults aged 18 to 60 years. 0.5-mL subcutaneous injection of 500 µg of adjuvanted (1 dose) or nonadjuvanted (2 doses) FLU-v (A-FLU-v or NA-FLU-v) or adjuvanted or nonadjuvanted placebo (A-placebo or NA-placebo) (2:2:1:1 ratio). Vaccine-specific cellular responses at days 0, 42, and 180 were assessed via flow cytometry and enzyme-linked immunosorbent assay. Solicited information on adverse events (AEs) was collected for 21 days after vaccination. Unsolicited information on AEs was collected throughout the study. The AEs with the highest incidence were mild to moderate injection site reactions. The difference between A-FLU-v and A-placebo in the median fold increase in secreted interferon-γ (IFN-γ) was 38.2-fold (95% CI, 4.7- to 69.7-fold; P = 0.001) at day 42 and 25.0-fold (CI, 5.7- to 50.9-fold; P < 0.001) at day 180. The differences between A-FLU-v and A-placebo in median fold increase at day 42 were 4.5-fold (CI, 2.3- to 9.8-fold; P < 0.001) for IFN-γ-producing CD4+ T cells, 4.9-fold (CI, 1.3- to 40.0-fold; P < 0.001) for tumor necrosis factor-α (TNF-α), 7.0-fold (CI, 3.5- to 18.0-fold; P < 0.001) for interleukin-2 (IL-2), and 1.7-fold (CI, 0.1- to 4.0-fold; P = 0.004) for CD107a. At day 180, differences were 2.1-fold (CI, 0.0- to 6.0-fold; P = 0.030) for IFN-γ and 5.7-fold (CI, 2.0- to 15.0-fold; P < 0.001) for IL-2, with no difference for TNF-α or CD107a. No differences were seen between NA-FLU-v and NA-placebo. The study was not powered to evaluate vaccine efficacy against influenza infection. Adjuvanted FLU-v is immunogenic and merits phase 3 development to explore efficacy. SEEK and the European Commission Directorate-General for Research and Innovation, European Member States within the UNISEC (Universal Influenza Vaccines Secured) project.


Gene expression patterns in human placenta.
The placenta is the principal metabolic, respiratory, excretory, and endocrine organ for the first 9 months of fetal life. Its role in fetal and maternal physiology is remarkably diverse. Because of the central role that the placenta has in fetal and maternal physiology and development, the possibility that variation in placental gene expression patterns might be linked to important abnormalities in maternal or fetal health, or even variations in later life, warrants investigation. As an initial step, we used DNA microarrays to analyze gene expression patterns in 72 samples of amnion, chorion, umbilical cord, and sections of villus parenchyma from 19 human placentas from successful full-term pregnancies. The umbilical cord, chorion, amnion, and villus parenchyma samples were readily distinguished by differences in their global gene-expression patterns, many of which seemed to be related to physiology and histology. Differentially expressed genes have roles that include placental trophoblast secretion, signal transduction, metabolism, immune regulation, cell adhesion, and structure. We found interindividual differences in expression patterns in villus parenchyma and systematic differences between the maternal, fetal, and intermediate layers. A group of genes that was expressed in both the maternal and fetal villus parenchyma sections of placenta included genes that may be associated with preeclampsia. We identified sets of genes whose expression in placenta was significantly correlated with the sex of the fetus. This study provides a rich and diverse picture of the molecular variation in the placenta from healthy pregnancies.


Appropriateness of proton pump inhibitor recommendations at hospital discharge and continuation in primary care.
Background: Inappropriate prescriptions of proton pump inhibitors (PPI) in hospital and primary care have been widely reported. Recommendations from hospital have been implicated as one reason for inappropriate prescriptions of PPI in primary care. Objective: To quantify the amount of appropriate PPI recommendations in hospital discharge letters and the influence of these recommendations on general practitioners' (GPs') PPI-prescriptions. Materials and Methods: This is an observational study in 31 primary care practices. We identified patients discharged from hospital with PPI recommendation between 2006 and 2007 and assessed practice records and PPI prescription six months prior and after hospital admission. Hospital recommendation for continuous PPI-treatment and continuation by GPs was classified as appropriate, inappropriate or uncertain. Logistic regression analysis was used to calculate factors associated with indicated and non-indicated PPI continuation. Results: In 263 (58%) out of 506 patients discharged from 35 hospitals with a PPI recommendation no indication could be found. Non-indicated PPIs were continued by GPs in 58% for at least 1 month. Indicated PPIs were discontinued in 33%. Two thirds of non-indicated PPIs were initiated in hospital. The strongest factor associated with non-indicated continuation was a PPI-prescription prior to hospital admission [OR: 3.0; 95% confidence interval (CI): 1.7-5.4]. This was also the strongest factor for continuation of an indicated PPI medication (OR: 3.2; 95% CI: 1.4-7.5). Conclusions: We found a strong influence of hospital recommendations and previous prescriptions on PPI prescriptions after discharge. Hospitals should critically review their practice of recommending PPI and document indications. GPs should carefully assess hospital recommendations and their medication prior to admission to avoid over- and under-prescribing.


Amantadine and dansylcadaverine inhibit vesicular stomatitis virus uptake and receptor-mediated endocytosis of alpha 2-macroglobulin.
The entry of many animal viruses into their host cells often proceeds via a specialized internalization pathway involving clathrin-coated regions of the plasma membrane. We have examined the effect of dansylcadaverine and amantadine on the entry of vesicular stomatitis virus (VSV) into mouse cells. Both compounds inhibit VSV entry as determined by fluorescence and electron microscopy, 3H-labeled VSV uptake, and VSV-dependent RNA synthesis assays. They also inhibit the uptake of alpha 2-macroglobulin, a protein that binds to specific membrane receptors and follows the same route of internalization. Dansylcadaverine is 20-fold more potent than amantadine in blocking virus and alpha 2-macroglobulin uptake. One cellular target for both of these amine-containing compounds appears to be the clustering of membrane-bound ligands or particles in clathrin-coated pits.


Interpreting changes in quality-of-life score in N of 1 randomized trials.
To provide additional evidence regarding the plausible range of the differences in health-related quality of life (HRQL) questionnaire scores within which the minimal important difference (MID) falls, we reviewed the results of 32 randomized controlled trials in individual subjects (N of 1 RCTs) with chronic diseases. These trials had been conducted to establish whether a patient was obtaining more good than harm from a medication. Each N of 1 RCT included a series of pairs of treatment periods, one period on active drug, and one on placebo or alternative drug. We examined the relationship between small (MID), medium, and large differences between periods within pairs, as indicated by Global Ratings and differences between these same periods according to HRQL questionnaires. The results showed a mean difference of 0.29 points per question in HRQL questionnaire scores corresponded to the MID. Differences of approximately 0.66 points per question corresponded to a moderate difference as ranked by the Global Rating; difference of about 1.09 points per question represented marked difference.


Arthrogenic muscle inhibition and return to sport after arthrofibrosis complicating anterior cruciate ligament surgery.
Arthrofibrosis is a devastating complication after Anterior Cruciate Ligament reconstruction (ACLr) characterized by a muscle weakness secondary to an arthrogenic muscle inhibition process. The loss of knee isokinetic strength due to arthrogenic muscle inhibition may be more important after arthrofibrosis, compared to an ACLr population with no complication. The isokinetic strength deficit [Limb Symmetry Index (LSI) at 60 and 180°/s of angular speed] was measured at 4, 7 and 12 post-operative months. Knee function, return to running and return to sport were evaluated. A comparison of the Quadriceps and the Hamstring LSI between patients with arthrofibrosis and those without post-operative complication was performed according to time and taking into consideration the type of surgical procedure. 539 primary ACLr patients were assessed. The arthrofibrosis group presented at 4, 7 and 12 post-operative months a Quadriceps LSI significantly lower compared to the control group, without influence of the graft procedure (LSI: 38, 53, 68% vs 63, 73, 85% at 60°/s, respectively). The Hamstring LSI was significantly lower at 4 and 7 post-operative months, but comparable at 12 months with an influence of the Hamstring procedure. Knee function was significantly lower at 4 and 7 post-operative months. Few arthrofibrosis ACLr patients returned to running at 7 post-operative months (6.8% vs 69.9%; p < .0001). An important and durable Quadriceps muscle weakness occurred after arthrofibrosis, whatever the type of graft procedure. This is explained by an Arthrogenic muscle inhibition which compromised the return to sport at the same level until 12 post-operative months.HighlightsAn important and durable quadriceps muscle weakness occurred after arthrofibrosis.The arthrogenic muscle inhibition was significant on the operated side independently of the type of the graft procedure.Arthrogenic muscle inhibition compromised the return to sport at the same level until 12 post-operative months.


Cutaneous Lyme borreliosis: Guideline of the German Dermatology Society.
This guideline of the German Dermatology Society primarily focuses on the diagnosis and treatment of cutaneous manifestations of Lyme borreliosis. It has received consensus from 22 German medical societies and 2 German patient organisations. It is the first part of an AWMF (Arbeitsgemeinschaft der Wissenschaftlichen Medizinischen Fachgesellschaften e.V.) interdisciplinary guideline: "Lyme Borreliosis - Diagnosis and Treatment, development stage S3". The guideline is directed at physicians in private practices and clinics who treat Lyme borreliosis. Objectives of this guideline are recommendations for confirming a clinical diagnosis, recommendations for a stage-related laboratory diagnosis (serological detection of IgM and IgG Borrelia antibodies using the 2-tiered ELISA/immunoblot process, sensible use of molecular diagnostic and culture procedures) and recommendations for the treatment of the localised, early-stage infection (erythema migrans, erythema chronicum migrans, and borrelial lymphocytoma), the disseminated early-stage infection (multiple erythemata migrantia, flu-like symptoms) and treatment of the late-stage infection (acrodermatitis chronica atrophicans with and without neurological manifestations). In addition, an information sheet for patients containing recommendations for the prevention of Lyme borreliosis is attached to the guideline.


Peptide and recombinant immunotherapy.
Because of the need to standardize allergen immunotherapy and the desire to reduce allergic adverse events during therapy, a transition to recombinant/synthetic hypoallergenic approaches is inevitable. Evidence supports the notion that effective therapy can be delivered using a limited panel of allergens or even epitopes, weakening the argument that all allergens must be present for optimal efficacy. Moreover, standardized products will allow direct comparisons between studies and, for the first time, immunotherapy studies will be truly blinded, allowing an accurate assessment of the actual treatment effect that can be achieved with this form of intervention.


Measuring the importance of different barriers to opioid agonist treatment using best-worst scaling in an Australian setting.
Opioid agonist treatment (OAT) is an effective treatment for opioid use disorder (OUD), however several client barriers to OAT are reported. Client importance of these barriers using economic preference elicitation measures have not been identified. This paper determines the most important OAT barriers using best-worst scaling (BWS) and compares the results of BWS to Likert scale. Cross-sectional self-completed survey with 191 opioid dependent clients who attended Australian needle and syringe sites. Participants were presented 15 Likert scale barriers and 15 BWS barrier scenarios. The BWS data was presented using count analysis, multinomial logit and mixed logit models. The ranking of barrier items was completed using three BWS methods and one Likert scale method, with share preference results (BWS) or mean scores (Likert) used to rank the 15 barriers. The most important client barriers were 'enjoy using opioids', 'lack of support services' and 'hard to access'. The four ranking methods produced different barrier rankings for the most important barriers, but similar results for the least important barriers. Policies around OAT as a harm reduction approach, increased support services and increased availability of OAT services would be beneficial in improving OAT uptake. Comparing BWS and Likert methods produced different highest ranked barriers, indicating the method used to identify preferences has significant implications on the type of intervention prioritised.


Human immunodeficiency virus transmission-Mechanisms underlying the cell-to-cell spread of human immunodeficiency virus.
Despite the success of combined antiretroviral therapy in controlling viral load and reducing the risk of human immunodeficiency virus (HIV) transmission, an estimated 1.5 million new infections occurred worldwide in 2021. These new infections are mainly the result of sexual intercourse and thus involve cells present on the genital mucosa, such as dendritic cells (DCs), macrophages (Mø) and CD4+ T lymphocytes. Understanding the mechanisms by which HIV interacts with these cells and how HIV exploits these interactions to establish infection in a new human host is critical to the development of strategies to prevent and control HIV transmission. In this review, we explore how HIV has evolved to manipulate some of the physiological roles of these cells, thereby gaining access to strategic cellular niches that are critical for the spread and pathogenesis of HIV infection. The interaction of HIV with DCs, Mø and CD4+ T lymphocytes, and the role of the intercellular transfer of viral particles through the establishment of the infectious or virological synapses, but also through membrane protrusions such as filopodia and tunnelling nanotubes (TNTs), and cell fusion or cell engulfment processes are presented and discussed.


Comparative study of toxicity of azo dye Procion Red MX-5B following biosorption and biodegradation treatments with the fungi Aspergillus niger and Aspergillus terreus.
Azo dyes are an important class of environmental contaminants and are characterized by the presence of one or more azo bonds (-N=N-) in their molecular structure. Effluents containing these compounds resist many types of treatments due to their molecular complexity. Therefore, alternative treatments, such as biosorption and biodegradation, have been widely studied to solve the problems caused by these substances, such as their harmful effects on the environment and organisms. The aim of the present study was to evaluate biosorption and biodegradation of the azo dye Procion Red MX-5B in solutions with the filamentous fungi Aspergillus niger and Aspergillus terreus. Decolorization tests were performed, followed by acute toxicity tests using Lactuca sativa seeds and Artemia salina larvae. Thirty percent dye removal of the solutions was achieved after 3 h of biosorption. UV-Vis spectroscopy revealed that removal of the dye molecules occurred without major molecular changes. The acute toxicity tests confirmed lack of molecular degradation following biosorption with A. niger, as toxicity to L. sativa seed reduced from 5% to 0%. For A. salina larvae, the solutions were nontoxic before and after treatment. In the biodegradation study with the fungus A. terreus, UV-Vis and FTIR spectroscopy revealed molecular degradation and the formation of secondary metabolites, such as primary and secondary amines. The biodegradation of the dye molecules was evaluated after 24, 240 and 336 h of treatment. The fungal biomass demonstrated considerable affinity for Procion Red MX-5B, achieving approximately 100% decolorization of the solutions by the end of treatment. However, the solutions resulting from this treatment exhibited a significant increase in toxicity, inhibiting the growth of L. sativa seeds by 43% and leading to a 100% mortality rate among the A. salina larvae. Based on the present findings, biodegradation was effective in the decolorization of the samples, but generated toxic metabolites, while biosorption was effective in both decolorization and reducing the toxicity of the solutions.


Distribution of HLA-B27 subtypes in patients with ankylosing spondylitis: the disease is associated with a common determinant of the various B27 molecules.
HLA-B27 subtypes can be defined by cellular, serological, and biochemical techniques. The seven subtypes so far identified represent structural variants of B27 with limited variations in the amino acid sequence of the B27 molecule. The routinely typed B27 'antigen' remains a common (shared, public) determinant present on various B27 molecules. The distribution of the subtypes varies strongly among different ethnic groups and they occur in different linkage disequilibria. In the healthy Dutch population only two subtypes were found: B27W (B27 X 1, B27M2+) (90%) and B27K (B27 X 2, B27M2-) (10%). A similar distribution of B27 subtypes was observed in 91 unrelated Dutch patients with ankylosing spondylitis (AS)--namely, 92% B27W and 8% B27K. In Oriental populations the subtype distribution is quite different: B27W occurs in less than 50%, whereas more than 50% individuals are of the B27C and B27D subtypes. Preliminary data indicate that the distribution of subtypes in healthy and diseased Oriental individuals is similar. These results suggest that the B27 and disease (AS) association is not correlated with the structural variations of one of the B27 subtypes, but with a common B27 determinant shared by various B27 subtypes. Consequently, the disease is older than the B27 variants. Further studies on disease and subtype distribution in various ethnic groups might contribute to a better understanding of the origin of both.


Investigation of center-specific saphenous vein utilization rates in femoral popliteal artery bypass and associated impact of conduit on outcomes.
The purpose of this study is to investigate variation in great saphenous vein (GSV) use among the various centers participating in the Vascular Quality Initiative infrainguinal bypass modules. Further, differences in outcomes in femoral-popliteal artery bypass with single segment GSV conduit vs prosthetic conduit will be documented. Center GSV use rate impact on outcomes will be investigated. Primary exclusions were patients undergoing redo bypass, urgent or emergent bypass, and those in whom prosthetic graft was used while having undergone prior coronary artery bypass grafting. The distribution of GSV use across the 260 centers participating in Vascular Quality Initiative infrainguinal bypass module was placed into histogram and variance in mean GSV use evaluated with analysis of variance analysis. Centers that used GSV in >50% of bypasses were categorized as high use centers and centers that used the GSV in <30% of cases were categorized as low use centers. Baseline differences in patient characteristics and comorbidities in those undergoing bypass with GSV vs prosthetic conduit were analyzed with χ2 testing and the Student t test, as were those undergoing treatment in high vs low use centers. Multivariable time-dependent Cox regression analyses were then performed for the primary outcomes of major amputation ipsilateral to the operative side and mortality in long-term follow-up. High vs low use center was a dichotomous variable in these regressions. Secondary outcomes of freedom from graft infection and freedom from loss of primary patency were performed with Kaplan-Meier analysis. Among centers with >50 patients meeting inclusion criteria for this study, GSV use ranged from 15% to 93% (analysis of variance P < .001). When considering all centers irrespective of number of patients, the range was 0% to 100%. On Kaplan-Meier analysis, GSV conduit use was associated with improved freedom from loss of primary or primary assisted patency, improved freedom from major amputation after index hospitalization, improved freedom from graft infection after the index hospitalization, and improved freedom from mortality in long-term follow-up (log-rank P < .001 for all four outcomes). Both low use center (hazard ratio, 1.35; P < .001) and prosthetic graft use (hazard ratio, 1.24; P < .001) achieved multivariable significance as risks for mortality in long-term follow-up. Other variables with a multivariable mortality association are presented in the manuscript. Low use center and prosthetic bypass were significant univariable but not multivariable risks for major amputation after index hospitalization. There is remarkably wide variation in GSV use for femoral popliteal artery bypass among various medical centers. GSV use is associated with enhanced long-term survival as well as freedom from loss of bypass patency and graft infection. The data herein indicate institutional patterns of prosthetic conduit choice, which has the potential to be altered to enhance outcomes.


Shc depletion stimulates brown fat activity in vivo and in vitro.
Adipose tissue is an important metabolic organ that integrates a wide array of homeostatic processes and is crucial for whole-body insulin sensitivity and energy metabolism. Brown adipose tissue (BAT) is a key thermogenic tissue with a well-established role in energy expenditure. BAT dissipates energy and protects against both hypothermia and obesity. Thus, BAT stimulation therapy is a rational strategy for the looming pandemic of obesity, whose consequences and comorbidities have a huge impact on the aged. Shc-deficient mice (ShcKO) were previously shown to be lean, insulin sensitive, and resistant to high-fat diet and obesity. We investigated the contribution of BAT to this phenotype. Insulin-dependent BAT glucose uptake was higher in ShcKO mice. Primary ShcKO BAT cells exhibited increased mitochondrial respiration; increased expression of several mitochondrial and lipid-oxidative enzymes was observed in ShcKO BAT. Levels of brown fat-specific markers of differentiation, UCP1, PRDM16, ELOVL3, and Cox8b, were higher in ShcKO BAT. In vitro, Shc knockdown in BAT cell line increased insulin sensitivity and metabolic activity. In vivo, pharmacological stimulation of ShcKO BAT resulted in higher energy expenditure. Conversely, pharmacological inhibition of BAT abolished the improved metabolic parameters, that is the increased insulin sensitivity and glucose tolerance of ShcKO mice. Similarly, in vitro Shc knockdown in BAT cell lines increased their expression of UCP1 and metabolic activity. These data suggest increased BAT activity significantly contributes to the improved metabolic phenotype of ShcKO mice.


First year after head and neck cancer: quality of life.
Treatment regimens for head and neck cancer patients profoundly affect several quality-of-life domains. Rehabilitative needs have been identified through cross-sectional analyses; however, few studies have prospectively assessed quality of life, included assessment of psychosocial variables, and identified predictors of long-term follow-up. The present study addresses these limitations through a prospective assessment of 105 patients with a newly diagnosed first primary squamous cell carcinoma of the oral cavity, pharynx, or larynx. Participants were enrolled onto a larger randomized controlled trial comparing a provider-delivered smoking cessation intervention with a usual-care-advice control condition. Participants completed a battery of self-report measures after diagnosis and before treatment and additional quality-of-life instruments at 1 and 12 months after initial smoking cessation advice. Participants displayed improvements at 12 months in functional status (P = .006) and in the areas of eating, diet, and speech; however, the latter three represent areas of continued dysfunction, and the changes were not statistically significant. Despite these improvements, patients reported a decline in certain quality-of-life domains, including marital (P = .002) and sexual functioning (P = .017), as well as an increase in alcohol use (P < .001). Predictors of quality of life at 12 months included treatment type, the Vigor subscale of the Profile of Mood States instrument, and quality-of-life scores obtained 1 month after initial smoking cessation advice. Results reinforce the need for rehabilitation management through the integration of psychologic and behavioral interventions in medical follow-up.


Headgroup and hydrocarbon tail effects on the surface tension of sugar-based surfactant solutions.
Measurements of surface tension isotherms were conducted for water solutions of pure and mixed n-decyl-beta-d-glucopyranoside (C(10)-Glu) and n-decyl-beta-d-maltopyranoside (C(10)-Mal) surfactants. By applying the Gibbs surface tension equation, the surface densities of Glu and Mal were derived for different compositions and concentrations. The surface fractions were compared with theoretically calculated values where the headgroups were modeled as hard disks. Satisfactory agreement was found for hard-disk sizes of 22.9 and 11.3 A(2) in the case of a 1:1 mixture. The results of the hard-disk calculations were employed to estimate the configurational free energy of the n-decyl-hydrocarbon chain. The results obtained agree well with previous calculations for the n-dodecyl chain. Comparison with n-dodecyl beta-d-maltopyranoside (C(12)-Mal) indicated a further contribution, with the longer hydrocarbon chain giving rise to a higher surface tension in good agreement with data for hydrocarbon liquids. Furthermore, the interpenetration of the headgroup into the hydrocarbon film was studied by means of comparing surface-tension data for n-decyl- and n-dodecyl-ethylene-oxide-based surfactants and n-decyl- and n-dodecyl-beta-d-thiomaltopyranosides (C(10)-S-Mal and C(12)-S-Mal, respectively) and -maltopyranosides. It was found that lengthening the tetra(etylene oxide) chain by one segment affects the surface tension only marginally, indicating little interpenetration of the additional ethylene-oxide group into the hydrocarbon film. For the thiomaltosides, however, the corresponding effect was found to be remarkably high.


[Congenital lens subluxation: visual acuity outcomes and intraocular lens postoperative position].
To evaluate the visual acuity outcomes and to investigate the intraocular lens (IOL) and endocapsular ring positions with ultrasound biomicroscopy in 17 eyes of 10 patients with congenital lens subluxation who underwent the same surgical technique, by the same surgeon. The study was performed in the ''Hospital de Olhos de Pernambuco'' and ''Fundação Altino Ventura''. The surgical technique consisted of phacoaspiration with implant of endocapsular ring and intraocular lens with one loop haptic amputated. The age varied from 7 to 22 years. Data on visual acuity (VA) before and after surgery, surgery follow-up period, and complications were analyzed. All patients underwent ultrasound biomicroscopy. The mean follow-up period was 2.8 years. There was a VA improvement in 17 (100%) eyes: in 12 eyes (70.6%) the visual acuity was better than 20/40; 4 (23.5%) ranged from 20/40 to 20/100, and 1 (5.9%) had visual acuity worse than 20/100, however better than the preoperative visual acuity. The posterior capsular opacification occurred in 10 eyes (58.9%). Ultrasound biomicroscopy showed that all IOL were partially decentralized, however without surpassing the pupil border limit. Endocapsular ring position was correct and there was a good capsular support in all cases. The evaluated surgical treatment provided good intraocular lens and endocapsular ring position, with VA improvement Thus, this technique is a viable, effective and safe option for the visual rehabilitation of patients with congenital lens subluxation.


Behavior of lambs in rest pens during long-distance transport.
The aim of this study was to determine how one group of lambs utilized 2 consecutive rest periods in novel environments with access to food and water that occurred during 22 hr of motor transport. The 18.5 +/- 0.6 kg lambs (n = 15) were transported for 8 hr and then unloaded for a 6-hr rest period. After 6 hr, the lambs were reloaded for another 8 hr of transport followed by a 24-hr rest period. Reloading for a second 8 hr of transport followed the initial rest period. The percentage of lambs engaged in drinking, eating, lying, playing, or "other" was determined at 15-min intervals. During the 6-hr rest period, peak lying behavior occurred during the 2nd and 6th hr of the period. During the first 6 hr of the 24-hr rest period, the percentage of lambs lying increased while the percentage of lambs eating decreased. In addition, the percentage of lambs lying during the first 6 hr of the 24-hr rest period was greater than during the 6-hr rest period. Lying down had a greater priority than eating during the second (24-hr) rest period.


Molecular cloning and characterization of gene encoding for cytoplasmic Hsc70 from Pennisetum glaucum may play a protective role against abiotic stresses.
Molecular chaperones (Hsps) have been shown to facilitate protein folding or assembly under various developmental and adverse environmental conditions. The aim of this study was to unravel a possible role of heat-shock proteins in conferring abiotic stress tolerance to plants. We isolated a cDNA encoding a cytoplasmic Hsp70 (PgHsc70) from Pennisetum glaucum by screening heat-stress cDNA library. PgHsc70 cDNA encoding 649 amino acids represents all conserved signature motifs characteristic of Hsp70s. The predicted molecular model of PgHsc70 protein suggests that the N-terminus ATP-binding region is evolutionarily conserved, in comparison to C-terminus peptide-binding domains. A single intron in ATPase domain coding region of PgHsc70 exhibited a high degree of conservation with respect to its position and phasing among other plant Hsp70 genes. Recombinant PgHsc70 protein purified from E. coli possessed in vitro chaperone activity and protected PgHsc70 expressing bacteria from damage caused by heat and salinity stress. Nucleotide sequence analysis of 5' flanking promoter region of PgHsc70 gene revealed a potential heat-shock element (HSE) and other putative stress-responsive transcription factor binding sites. Positive correlation existed between differentially up-regulated PgHsc70 transcript levels and the duration and intensity of different environmental stresses. Molecular and biochemical analyses revealed that PgHsc70 gene was a member of the Hsp70 family and suggested that its origin was from duplication of a common ancestral gene. Transcript induction data, presence of several putative stress-responsive transcription factor-binding sites in the promoter region of PgHsc70 and the presence of a protective in vitro chaperone activity of this protein against damage caused by heat and salinity, when expressed in E. coli, suggest its probable role in conferring abiotic stress tolerance to this plant.


Spatial covariation in nutrient enrichment and fishing of herbivores in an oceanic coral reef ecosystem.
Both natural and anthropogenic stressors are increasing on coral reefs, resulting in large-scale loss of coral and potential shifts from coral- to macroalgae-dominated community states. Two factors implicated in shifts to macroalgae are nutrient enrichment and fishing of reef herbivores. Although either of these factors alone could facilitate establishment of macroalgae, reefs may be particularly vulnerable to coral-to-algae phase shifts in which strong bottom-up forcing from nutrient enrichment is accompanied by a weakening of herbivore control of macroalgae via intense fishing. We explored spatial heterogeneity and covariance in these drivers on reefs in the lagoons of Moorea, French Polynesia, where the local fishery heavily targets herbivorous fishes and there are spatially variable inputs of nutrients from agricultural fertilizers and wastewater systems. Spatial patterns of fishing and nutrient enrichment were not correlated at the two landscape scales we examined: among the 11 interconnected lagoons around the island or among major habitats (fringing reef, mid-lagoon, back reef) within a lagoon. This decoupling at the landscape scale resulted from patterns of covariation between enrichment and fishing that differed qualitatively between cross-shore and long-shore directions. At the cross-shore scale, nutrient enrichment declined but fishing increased from shore to the crest of the barrier reef. By contrast, nutrient enrichment and fishing were positively correlated in the long-shore direction, with both increasing with proximity to a pass in the barrier reef. Contrary to widespread assumptions in the scientific literature that human coastal population density correlates with impact on marine ecosystems and that fishing effort declines linearly with distance from the shore, these local stressors produced a complex spatial mosaic of reef vulnerabilities. Our findings support spatially explicit management involving the control of anthropogenic nutrients and strategic reductions in fishing pressure on herbivores by highlighting specific areas to target for management actions.


Copper(II) complexes as turn on fluorescent sensors for nitric oxide.
Two copper(II) complexes, 1 and 2, of two tridentate N-donor ligands, L(1) and L(2) [L(1) = dansyl derivative of bis-[3-(dimethylamino)-propyl]amine; L(2) = dansyl derivative of dipropylenetriamine] were synthesized and characterized. The quenched fluorescence intensity of complexes 1 and 2, in degassed methanol or aqueous (buffered at pH 7.2) solution, was found to reappear on exposure to nitric oxide. This is attributed to the reduction of paramagnetic Cu(II) center by nitric oxide to diamagnetic Cu(I).


p16 Expression and Biological Behavior of Flat Vulvar Low-grade Squamous Intraepithelial Lesions (LSIL).
Flat low-grade squamous intraepithelial lesion (LSIL) of the vulva [vulvar intraepithelial neoplasia (VIN) 1, flat condyloma] is an uncommon entity with poorly understood biological behavior. We aimed to determine the risk of subsequent vulvar high-grade squamous intraepithelial lesion (HSIL) or carcinoma following a diagnosis of vulvar LSIL/VIN 1, as well as the frequency and predictive value of p16 immunohistochemical expression in this setting. Of the 51 included cases, p16 positivity (diffuse block staining) was identified in 2 (4%). Follow-up data were available in 34 cases, of which 2 (5.9%) developed subsequent vulvar HSIL, including 1/2 p16-positive cases and 1/32 p16-negative cases. The difference in HSIL frequency between p16-positive and p16-negative cases was not statistically significant (P=0.116 for VIN 2+, P=0.061 for VIN 3). For the 18 patients with treatment information available, 10 (56%) received medical or surgical treatment after biopsy. Our results indicate that flat vulvar LSIL is infrequently p16 positive, and that few patients with vulvar LSIL develop subsequent vulvar HSIL. Despite the use of destructive treatment in some cases, the data provide support for the nonpreneoplastic nature of the entity. Immunohistochemical expression of p16 may not be a predictor of HSIL risk in vulvar LSIL, although this result may also be related to the very low rates of both p16 positivity and subsequent vulvar HSIL in our sample. It is clear that vulvar LSIL is distinct from LSIL in other lower anogenital sites in terms of its behavior and p16 expression frequency.


Polarized light through polycrystalline vaterite helicoids.
Vaterite helicoids [W. Jiang et al., Nat. Commun., 2017, 8, 15066] are chiral, polycrystalline suprastructures grown in the presence of the amino acids, aspartic (Asp) or glutamic (Glu) acid, that are abundant in proteins regulating biomineralization. These complex objects are composed of hexagonal vaterite nanocrystals assembled as curved-edge platelets that form chiral ensembles. The sense stacked platelets is correlated with the stereochemistry of the amino acid additive: l-Asp gives counterclockwise architectures while d-Asp gives the clockwise enantiomorphs. As new layers stack, platelets become progressively inclined with respect to the substrate suface. The growth and structure of vaterite helicoids was originally evidenced by electron microscopy and atomic force microscopy. Here, we develop an optical model for describing polarized light transmission through helicoids as measured by Mueller matrix polarimetry. The close agreement between experimental measurements and simulation confirms that the propellor-like organization of inclined platelets creates optically active structures determined by growth additive stereochemistry. The microscopy employed demonstrates the information that can be obtained by complete polarimetry using a camera as a light detector, a technique that could be applied profitably to all manner of complex structures organized from anisotropic particles.


Delayed-onset sinus node dysfunction in a child victim of Russell's viper bite.
Cardiac complications following envenomation by Russell's viper venom are uncommon. We describe a 14-year-old girl who developed delayed-onset sinus node dysfunction. She presented with mucosal bleeding, ptosis, and muscle weakness. Her 20-min whole blood clotting time and international normalized ratio were prolonged. The initial electrocardiogram showed sinus tachycardia. Her systemic manifestations responded to antivenom serum. After 24 h, she developed bradycardia and electrocardiography showed sinus node dysfunction with sinus arrest and an atrial escape rhythm. This case shows that arrhythmias can have a delayed onset even after resolution of other systemic manifestations, and even after treatment with antivenom serum.


A focus on the biosynthesis and composition of cuticle in fruits.
Cuticles are plant structures, composed mostly by lipidic layers, synthesized by nonwoody aerial plant organs and deposited on the surface of outer epidermal cell walls. Although its significance has been often disregarded, cuticle deposition modifies organ chemistry, influences mechanical properties, and plays a central role in sensing and interacting with the surrounding environment. Even though some research has been undertaken addressing cuticle biosynthesis and composition in vegetative plant tissues, comparatively less information is available regarding cuticle composition in the epidermis of fruits. However, recent work points to a role for cuticles in the modulation of fruit quality and postharvest performance, indicating that current models for the investigation of fruit development, metabolism, and quality need to integrate a comprehensive knowledge of the cuticle layer. This paper provides an overview of recent findings and observations regarding cuticle biosynthesis and composition in fruits from species of agronomic and economic relevance. Important, but often neglected differences in cuticle composition and biosynthesis patterns among diverse fruit species are described herein to generate an atlas of what is currently known about fruit cuticles and to highlight what remains to be explored. Emphasis is placed on the need to investigate each genetic background considering its own specificities, to permit correlations with the particular physiology of each species considered. Both specific composition and changes during maturation and ripening are reviewed.


[A new echographic sign for the reliable diagnosis of graves' disease (author's transl)].
With standardized echography, an ultrasonic method specifically designed for tissue differentiation, Graves' orbitopathy can be diagnosed reliably and accurately even when the clinical picture and thyroid tests are not typical for this condition (e.g., with unilateral exophthalmus, lack of lid signs, euthyroid status). A new echographic criterion of Graves' orbitopathy, i.e., the heterogeneous and high-reflective structure of the diseased muscle tissue, plays a key role in the differential diagnosis.


Species richness on coral reefs and the pursuit of convergent global estimates.
Global species richness, whether estimated by taxon, habitat, or ecosystem, is a key biodiversity metric. Yet, despite the global importance of biodiversity and increasing threats to it (e.g., we are no better able to estimate global species richness now than we were six decades ago. Estimates of global species richness remain highly uncertain and are often logically inconsistent. They are also difficult to validate because estimation of global species richness requires extrapolation beyond the number of species known. Given that somewhere between 3% and >96% of species on Earth may remain undiscovered, depending on the methods used and the taxa considered, such extrapolations, especially from small percentages of known species, are likely to be highly uncertain. An alternative approach is to estimate all species, the known and unknown, directly. Using expert taxonomic knowledge of the species already described and named, those already discovered but not yet described and named, and those still awaiting discovery, we estimate there to be 830,000 (95% credible limits: 550,000-1,330,000) multi-cellular species on coral reefs worldwide, excluding fungi. Uncertainty surrounding this estimate and its components were often strongly skewed toward larger values, indicating that many more species on coral reefs is more plausible than many fewer. The uncertainties revealed here should guide future research toward achieving convergence in global species richness estimates for coral reefs and other ecosystems via adaptive learning protocols whereby such estimates can be tested and improved, and their uncertainties reduced, as new knowledge is acquired.


Perioperative Hypothermia Is Associated With Increased 30-Day Mortality in Hip Fracture Patients in the United Kingdom: Α Systematic Review and Meta-analysis.
To undertake a systematic review and meta-analysis to determine the relationship between perioperative hypothermia and mortality after surgery for hip fracture. A systematic literature search of Medline, EMBASE, CINAHL, and Cochrane CENTRAL databases was performed using the Cochrane methodology for systematic reviews with no publication year limit. Only studies available in the English language were included. Predetermined inclusion criteria were patients of any age with a hip fracture, exposure was their body temperature and outcome was mortality rate. Any comparative study design was eligible. The quality of selected studies was assessed according to each study design with the Methodological Index for Non-Randomised Studies (MINORS) used for all the retrospective comparative studies. The GRADE approach was used to assess the quality of evidence. A meta-analysis was conducted using a random-effects model. The literature search identified 1016 records. After removing duplicates and those not meeting inclusion criteria, 3 studies measuring 30-day mortality were included. All included studies were carried out in the United Kingdom. The mortality rate was higher in the hypothermic groups as compared with the normothermic group in all the studies, with the difference being significant in 2 of the studies (P < 0.0001). The meta-analysis showed that low body temperature was associated with an increased mortality risk (estimated odds ratio: 2.660; 95% confidence interval: 1.948-3.632; P < 0.001) in patients undergoing surgery for hip fracture. This study shows that low body temperature in hip fracture patients is associated with an increased 30-day mortality risk in the United Kingdom. Randomized control trials are required to determine whether the association between perioperative hypothermia in hip fracture patients and mortality is causal. Nevertheless, based on this analysis, we urge the maintenance of normal body temperature in the perioperative period to be included in national hip fracture guidelines. Prognostic Level III. See Instructions for Authors for a complete description of levels of evidence.


Corticosteroids for parasitic eosinophilic meningitis.
Angiostrongylus cantonensis (A. cantonensis) is the major cause of infectious eosinophilic meningitis. Dead larvae of this parasite cause inflammation and exacerbate symptoms of meningitis. Corticosteroids are drugs used to reduce the inflammation caused by this parasite. To assess the efficacy and safety of corticosteroids for the treatment of eosinophilic meningitis. We searched CENTRAL (2014, Issue 11), MEDLINE (1950 to November Week 3, 2014), EMBASE (1974 to December 2014), Scopus (1960 to December 2014), Web of Science (1955 to December 2014), LILACS (1982 to December 2014) and CINAHL (1981 to December 2014). Randomised controlled trials (RCTs) of corticosteroids versus placebo for eosinophilic meningitis. Two review authors (SiT, SaT) independently collected and extracted study data. We graded the methodological quality of the RCTs. We identified and analysed outcomes and adverse effects. We did not identifiy any new trials for inclusion or exclusion in this 2014 update. One study involving 110 participants (55 participants in each group) met our inclusion criteria. The corticosteroid (prednisolone) showed a benefit in shortening the median time to resolution of headaches (five days in the treatment group versus 13 days in the control group, P value < 0.0001). Corticosteroids were also associated with smaller numbers of participants who still had headaches after a two-week course of treatment (9.1% versus 45.5%, P value < 0.0001). The number of patients who needed repeat lumbar puncture was also smaller in the treatment group (12.7% versus 40%, P value = 0.002). There was a reduction in the median time of analgesic use in participants receiving corticosteroids (10.5 versus 25.0, P value = 0.038). There were no reported adverse effects from prednisolone in the treatment group. Corticosteroids significantly help relieve headache in patients with eosinophilic meningitis, who have a pain score of four or more on a visual analogue scale. However, there is only one RCT supporting this benefit and this trial did not clearly mention allocation concealment and stratification. Therefore, we agreed to grade our included study as a moderate quality trial. Future well-designed RCTs are necessary.


Suppressive effects of ascorbate derivatives on ultraviolet-B-induced injury in HaCaT human keratinocytes.
The aging of skin, including sunburning, is caused by ultraviolet (UV) irradiation. Here, we examined the inhibitory effect of ascorbic acid (AsA) and its derivatives AsA 2-phosphate (AA-2P) and AsA 2-glucoside (AA-2G) on UV-B- induced cytotoxicity in HaCaT keratinocytes. Results show that cell viability significantly decreased when exposed to UV-B at 0.1-0.4 J/cm2 in a dose-dependent manner. In this study, AsA could not inhibit cytotoxicity, but AA-2P and AA-2G was able to cancel the harmful effect of UV-B when treated at high levels of 0.5-5 mM. These results indicate that the masking of the C-2 OH group may be an effective modification for AsA to inhibit UV-B-induced cytotoxicity in human keratinocytes.


MRI T2 and T1ρ relaxation in patients at risk for knee osteoarthritis: a systematic review and meta-analysis.
Magnetic resonance imaging (MRI) T2 and T1ρ relaxation are increasingly being proposed as imaging biomarkers potentially capable of detecting biochemical changes in articular cartilage before structural changes are evident. We aimed to: 1) summarize MRI methods of published studies investigating T2 and T1ρ relaxation time in participants at risk for but without radiographic knee OA; and 2) compare T2 and T1ρ relaxation between participants at-risk for knee OA and healthy controls. We conducted a systematic review of studies reporting T2 and T1ρ relaxation data that included both participants at risk for knee OA and healthy controls. Participant characteristics, MRI methodology, and T1ρ and T2 relaxation data were extracted. Standardized mean differences (SMDs) were calculated within each study. Pooled effect sizes were then calculated for six commonly segmented knee compartments. 55 articles met eligibility criteria. There was considerable variability between scanners, coils, software, scanning protocols, pulse sequences, and post-processing. Moderate risk of bias due to lack of blinding was common. Pooled effect sizes indicated participants at risk for knee OA had lengthened T2 relaxation time in all compartments (SMDs from 0.33 to 0.74; p < 0.01) and lengthened T1ρ relaxation time in the femoral compartments (SMD from 0.35 to 0.40; p < 0.001). T2 and T1ρ relaxation distinguish participants at risk for knee OA from healthy controls. Greater standardization of MRI methods is both warranted and required for progress towards biomarker validation.


Update on the management of COPD.
COPD is highly prevalent and will continue to be an increasing cause of morbidity and mortality worldwide. COPD is now viewed under a new paradigm as preventable and treatable. In addition, it has become accepted that COPD is not solely a pulmonary disease but also one with important measurable systemic consequences. Patients with COPD have to be comprehensively evaluated to determine the extent of disease so that therapy can be adequately individualized. We now know that smoking cessation, oxygen for hypoxemic patients, lung reduction surgery for selected patients with emphysema, and noninvasive ventilation during severe exacerbations have an impact on mortality. The completion of well-planned pharmacologic trials have shown the importance of decreasing resting and dynamic hyperinflation on patient-centered outcomes and the possible impact on mortality and rate of decline of lung function. In addition, therapy with pulmonary rehabilitation and lung transplantation improve patient-centered outcomes such as health-related quality of life, dyspnea, and exercise capacity. Rational use of single or multiple therapeutic modalities in combination have an impact on exacerbations and hospitalizations. This monograph presents an integrated approach to patients with COPD and updates their management incorporating the recent advances in the field. The future for patients with COPD is bright as primary and secondary prevention of smoking becomes more effective and air quality improves. In addition, current research will unravel the pathogenesis, clinical, and phenotypic manifestations of COPD, thus providing exciting therapeutic targets. Ultimately, the advent of newer and more effective therapies will lead to a decline in the contribution of this disease to poor world health.


Direct Evidence for Li Ion Hopping Conduction in Highly Concentrated Sulfolane-Based Liquid Electrolytes.
We demonstrate that Li+ hopping conduction, which cannot be explained by conventional models i.e., Onsager's theory and Stokes' law, emerges in highly concentrated liquid electrolytes composed of LiBF4 and sulfolane (SL). Self-diffusion coefficients of Li+ ( DLi), BF4- ( DBF4), and SL ( DSL) were measured with pulsed-field gradient NMR. In the concentrated electrolytes with molar ratios of SL/LiBF4 ≤ 3, the ratios DSL/ DLi and DBF4/ DLi become lower than 1, suggesting faster diffusion of Li+ than SL and BF4-, and thus the evolution of Li+ hopping conduction. X-ray crystallographic analysis of the LiBF4/SL (1:1) solvate revealed that the two oxygen atoms of the sulfone group are involved in the bridging coordination of two different Li+ ions. In addition, the BF4- anion also participates in the bridging coordination of Li+. The Raman spectra of the highly concentrated LiBF4-SL solution suggested that Li+ ions are bridged by SL and BF4- even in the liquid state. Moreover, detailed investigation along with molecular dynamics simulations suggests that Li+ exchanges ligands (SL and BF4-) dynamically in the highly concentrated electrolytes, and Li+ hops from one coordination site to another. The spatial proximity of coordination sites, along with the possible domain structure, is assumed to enable Li+ hopping conduction. Finally, we demonstrate that Li+ hopping suppresses concentration polarization in Li batteries, leading to increased limiting current density and improved rate capability compared to the conventional concentration electrolyte. Identification and rationalization of Li+ ion hopping in concentrated SL electrolytes is expected to trigger a new paradigm of understanding for such unconventional electrolyte systems.


Dentin regeneration using deciduous pulp stem/progenitor cells.
Reparative dentin formation is essential for maintaining the integrity of dentin structure during disease or trauma. In this study, we investigated stem/progenitor cell-based tissue engineering for dentin regeneration in a large animal model. Porcine deciduous pulp stem/progenitor cells (PDPSCs) were mixed with a beta-tricalcium phosphate (β-TCP) scaffold for dentin regeneration. Different concentrations of PDPSCs were tested to determine the optimal density for dentin regeneration. Aliquots of 5×10(5) PDPSCs in 1 mL resulted in the highest number of cells attached to the scaffold and the greatest alkaline phosphatase activity. We labeled PDPSCs with green fluorescent protein (GFP) and used the optimal cell numbers mixed with β-TCP to repair pulp chamber roof defects in the premolars of swine. Four weeks after transplantation, GFP-positive PDPSCs were observed in PDPSC-embedded scaffold constructs. At 16 weeks after transplantation, the PDPSCs mixed with β-TCP significantly regenerated the dentin-like structures and nearly completely restored the pulp chamber roof defects. This study demonstrated that the PDPSC/scaffold construct was useful in direct pulp-capping and provides pre-clinical evidence for stem/progenitor cell-based dentin regeneration.


CYP1B1 gene mutations causing primary congenital glaucoma in Tunisia.
Primary congenital glaucoma (PCG) is responsible for a significant proportion of childhood blindness in Tunisia. Early prevention based on genetic diagnosis is therefore required. This study sought to determine the frequency of CYP1B1 (cytochrome P450, family 1, subfamily B, polypeptide 1) mutations in 18 PCG patients, recruited from Central and Southern of Tunisia. Genomic DNA was extracted and the coding regions of CYP1B1 were analysed by direct sequencing. A phylogenetic network of CYP1B1 haplotypes was drawn using the median-joining algorithm. Sequence analysis revealed a "tetra-allelic mutation" (two novel mutations, p.F231I and p.P437A in the homozygous state) in one patient. The healthy members of his family carried those variations on the same allele. Two previously described mutations p.G61E and c.535delG were also identified in the homozygous state in seven and two probands, respectively. Seven single-nucleotide polymorphisms were identified and used to generate haplotypes. Our results showed that the CYP1B1 mutations were present in 55% of Tunisian PCG patients' alleles. Haplotype analysis allowed us to define the proto-haplotype and to confirm historical migratory flows. Establishment of PCG genetic aetiology in Tunisia will improve genetic diagnosis and counselling.


Parent Contributions to the Development of Political Attitudes in Adoptive and Biological Families.
Where do our political attitudes originate? Although early research attributed the formation of such beliefs to parent and peer socialization, genetically sensitive designs later clarified the substantial role of genes in the development of sociopolitical attitudes. However, it has remained unclear whether parental influence on offspring attitudes persists beyond adolescence. In a unique sample of 394 adoptive and biological families with offspring more than 30 years old, biometric modeling revealed significant evidence for genetic and nongenetic transmission from both parents for the majority of seven political-attitude phenotypes. We found the largest genetic effects for religiousness and social liberalism, whereas the largest influence of parental environment was seen for political orientation and egalitarianism. Together, these findings indicate that genes, environment, and the gene-environment correlation all contribute significantly to sociopolitical attitudes held in adulthood, and the etiology and development of those attitudes may be more important than ever in today's rapidly changing sociopolitical landscape.


Gene expression of collagen types IIA and IX correlates with ultrastructural events in early osteoarthrosis: new applications of the rabbit meniscectomy model.
To examine the phenotypic expression and geographic distribution of collagens in early stages of osteoarthrosis and their relationship to ultrastructural events in cartilage. In situ hybridization was used to localize articular expression of total type II (A+B) and type IX collagen at 2 and 4 weeks in the rabbit meniscectomy model of osteoarthrosis. The expression of the developmental marker collagen IIA was analyzed at the same time points. Articular cartilage structure was examined by scanning electron microscopy. Little difference was found in total type II or type IX collagen gene expression for operated versus control limbs at 2 weeks. Gene expression for collagen types IIA and IX was found to be site-specific by the 4 week period and was largely limited to the meniscectomy site. At 4 weeks, this activity was correlated with site-specific alterations in chondrocyte morphology, qualitative changes in the collagen matrix, and articular surface delamination on microscopy. Gene expression for collagen types IIA and IX is site-specific and correlates with ultrastructural changes in cartilage in this model of early osteoarthrosis. We present the first known report of the distribution of type IX collagen gene expression in any model of osteoarthrosis. These findings support the central importance of matrix interactions in osteoarthrosis and suggest that early phases of repair involve re-expression of a developmental sequence by chondrocytes.


Effects of different forms of environmental enrichment on behavioral, endocrinological, and immunological parameters in male mice.
This study investigated effects of different forms of environmental enrichment on behavioral, endocrinological, and immunological parameters in male mice. For this purpose, animals of the inbred strain CS were kept in groups of four males under three different housing conditions: (A) nonstructured Makrolon type III laboratory cages ("standard-housing" = S); (B) equivalent laboratory cages that were enriched with a box and a scaffolding ("enriched-housing" = E); and (C) spacious terraria that were structured richly ("super-enriched-housing" = SE). Both forms of enrichment caused a sharp rise in aggressive behavior, though play behavior was increased in E and SE mice, too. Levels of sociopositive behaviors in S and SE mice were higher than those in E mice. Plasma corticosterone concentrations and adrenal tyrosine hydroxylase activities were significantly increased in male mice kept in both forms of enriched cages, indicating an activation of the pituitary-adrenocortical and the adrenomedullary systems. The behavioral and endocrinological differences were partly reflected by immunological parameters: SE mice had levels of IgG1 and ratios of IFN-gamma/IL-10 and IL-2/IL-10 significantly lower than those of S mice. Ratios of IgG2a/IgG1 were significantly higher in SE mice. The absolute percentages of CD8 cells in E-mice were significantly lower than those in S mice. Despite the elevated levels of stress hormones under both forms of enriched housing, the behavioral parameters also indicate positive effects of the enrichment, especially on SE animals. Obviously, an environmental enrichment is beneficial for male mice as long as the spatial conditions are generous enough to allow coping with the increased aggression brought about by the enrichment.


Intraperitoneal application of bupivacaine plus morphine for pain relief after laparoscopic cholecystectomy.
Intraperitoneal administration of a local anaesthetic in combination with an opioid, for the relief of postoperative pain, has already been reported except after laparoscopic cholecystectomy. This study was aimed at assessing the analgesic effect of the intraperitoneal administration of bupivacaine and morphine in patients undergoing laparoscopic cholecystectomy. At the end of laparoscopic cholecystectomy, in a double-blind, randomized manner, one of the following injections was given intraperitoneally. There were 30 patients in each group: Group 1, physiological saline 30 mL; Group 2, bupivacaine 0.25% 30 mL; Group 3, bupivacaine 0.25% 30 mL plus morphine 2 mg. In addition, Group 2 received 2 mg intravenous (i.v.) morphine in 2 mL saline, and Groups 1 and 3, 2 mL saline intravenously. Patients' postoperative pain was evaluated using a visual analogue scale and a verbal rating score. The postoperative analgesic requirement was assessed by the total dose of metamizol administered by an i.v. patient-controlled analgesia (PCA) device. Pain, vital signs, supplemental analgesic consumption and side-effects were recorded for all patients for 24 h. There were no differences between the three groups regarding pain scores (at rest and coughing) during the study except in the first 2 h, when scores were lower for patients receiving intraperitoneal bupivacaine plus i.v. morphine (P < 0.05). Supplemental consumption of metamizol was significantly lower (P < 0.05) in Group 3 than in Group 1 during the first 6 h after surgery. However, the cumulative doses of metamizol were also lower in Group 2 than in Groups 1 and 3 over the entire study (2025 +/- 1044 mg vs. 4925 +/- 1238 and 4125 +/- 1276mg; P < 0.05). In patients undergoing laparoscopic cholecystectomy, the intraperitoneal administration of morphine plus bupivacaine 0.25% reduced the analgesic requirements during the first 6 postoperative hours compared with the control group. However, the combination of intraperitoneal bupivacaine 0.25% and i.v. morphine was more effective for treatment of pain after laparoscopic cholecystectomy.


Divergent Neuroimmune Signatures in the Cerebrospinal Fluid Predict Differential Gender-Specific Survival Among Patients With HIV-Associated Cryptococcal Meningitis.
Survival among people with HIV-associated cryptococcal meningitis (CM) remains low, exceptionally among women with the increased threat of death on current optimal use of antifungal drugs. Cryptococcus dissemination into the central nervous system (CNS) prompts a neuroimmune reaction to activate pathogen concomitant factors. However, no consistent diagnostic or prognostic immune-mediated signature is reported to underpin the risk of death or mechanism to improve treatment or survival. We theorized that the distinct neuroimmune cytokine or chemokine signatures in the cerebrospinal fluid (CSF), distinguish survivors from people who died on antifungal treatment, who may benefit from tailored therapy. We considered the baseline clinical disease features, cryptococcal microbiologic factors, and CSF neuroimmune modulated signatures among 419 consenting adults by gender (biological sex assigned at birth) (168 females and 251 males) by 18 weeks of survival on antifungal management. Survival at 18 weeks was inferior among females than males (47% vs. 59%; hazard ratio HR=1.4, 95% CI: 1.0 to 1.9, and p=0.023). Unsupervised principal component analysis (PCA) demonstrated the divergent neuroimmune signatures by gender, survival, and intragender-specific survival. Overall, females displayed lower levels of PD-L1, IL-1RA, and IL-15 than males (all p≤0.028). Female survivors compared with those who died, expressed significant fold elevations in levels of CSF (CCL11 - myeloid and CXCL10 - lymphoid chemokine (in both p=0.001), and CSF Th1, Th2, and Th17 cytokines. In contrast, male survivors expressed distinctly lower levels of CSF IL-15 and IL-8 compared with those who died. Survivors of either gender demonstrated a significant increase in the levels of immune regulatory element, IL-10. In the finale, we classified divergent neuroimmune key signatures in CSF by gender, survival, and intragender-specific survival among people with HIV-associated cryptococcal meningitis. These intragender-specific survival associated-neuroimmune signatures, suggests the discrete role of gender immune regulating mechanisms as the possible targets for interventions to advance therapy to improve survival among people with HIV-associated cryptococcal meningitis.


Protection against cisplatin-induced ototoxicity by N-acetylcysteine in a rat model.
Cisplatin (CDDP) is a widely used chemotherapeutic agent that is highly ototoxic. Animal studies and clinical trials have shown that thiosulfates can protect against platinum-induced ototoxicity. This study investigated a new model for CDDP ototoxicity in the rat, and tested the potential chemoprotective effect of administering N-acetylcysteine (NAC) before giving CDDP. Long Evans rats were treated with CDDP 6 mg/kg delivered to the aorta via a retrograde right external carotid artery infusion, 15 min after intravenous (IV) infusion of saline (n=8) or NAC 400 mg/kg (n=8), such that the vertebral arteries were perfused. Subsequent groups were similarly treated with NAC 30 min before (n=7) and 4 h after (n=7) CDDP. Auditory brainstem response (ABR) thresholds were tested at 4-20 kHz, 7 days after treatment and compared to baseline ABR values. The NAC-treated rats exhibited no significant change from baseline values at all time intervals, while the saline-treated rats showed marked ototoxicity, especially at higher frequencies. Furthermore, the rats treated with NAC 15 min before CDDP exhibited less overall toxicity to CDDP, as evidenced in weight loss 7 days post-treatment (mean for saline=-39.63 g; mean for NAC=-21.13 g; p=0.0084). These data show that treatment with NAC can prevent CDDP-induced ototoxicity in rats.


Temporal changes in surface-water insecticide concentrations after the phaseout of diazinon and chlorpyrifos.
The recent (late 2001) federally mandated phaseout of diazinon and chlorpyrifos insecticide use in outdoor urban settings has resulted in a rapid decline in concentrations of these insecticides in urban streams and rivers in the northeastern and midwestern United States. Assessment of temporal insecticide trends at 20 sites showed that significant step decreases in diazinon concentrations occurred at 90% of the sites after the phaseout, with concentrations generally decreasing by over 50% in summer samples. Chlorpyrifos concentrations showed significant step decreases in at least 1 season at 3 of the 4 sites with sufficient data for analysis. The decrease in diazinon concentrations in response to the phaseout resulted in a decline in the frequency of concentrations exceeding the acute invertebrate water-quality benchmark of 0.1 microg/L from 10% of pre-phaseout summer samples to fewer than 1% of post-phaseout summer samples. Although some studies have indicated an increase in concentrations of carbaryl in response to the organophosphorous phaseout, carbaryl concentrations only increased at 1 site after the phaseout. A full assessment of the effect of the phaseout of diazinon and chlorpyrifos on surface water will require data on other insecticides used to replace these compounds.


Factors associated with progression to pre-diabetes: a recurrent events analysis.
Pre-diabetes is a strong risk factor for type 2diabetes (T2D). The aim of this study was to explore factors associated with normal glucose maintenance and pre-diabetes prevention or delay. Data of 1016 first-degree relatives of T2D patients were retrieved from the Isfahan Diabetes Prevention Study (IDPS). Association of various variables including nutrients, serum tests and physical activity with the risk of pre-diabetes was assessed using recurrent events approach. Cumulative incidence of diabetes was 8.17, 9.44, and 4.91% for total sample and individuals with and without pre-diabetes experience in the follow-up. Risk of progression to pre-diabetes was higher in women and older people (p < 0.01). Additionally, BMI and blood pressure had significant association with the risk (p < 0.01) and individuals with higher intake of fat were at higher risk (HR = 2.26; 95% CI 1.66-3.07 for high-intake and HR = 1.52; 95% CI 1.27-1.83 for medium-intake compared to low-intake group). Carbohydrates and protein intake were positively associated with the risk of pre-diabetes with HR = 8.63 per 49 g extra carbohydrates per day and HR = 1.32 per 6 g extra protein per day (p < 0.01). The association was also significant for triglyceride (TG) with 7% risk increase per 1 SD = 1.14 increase in TG level. Despite frequent studies on lifestyle modification for pre-diabetes prevention, less information is available about the role of nutritional components. We observed direct effects for intake of macronutrients including fat, carbohydrates, and protein in first-degree relatives. Further research is warranted to assess these associations in general populations. Level III: Evidence obtained from a single-center cohort study.