Deep Learning Based Metabolite Annotation.
Metabolite annotation is a major bottleneck in untargeted metabolomics studies by liquid chromatography coupled with mass spectrometry (LC-MS). This is in part due to the limited publicly available spectral libraries, which consist of tandem mass spectrometry (MS/MS) data acquired from just a fraction of known compounds. Machine learning and deep learning methods provide the opportunity to predict molecular fingerprints based on MS/MS data. The predicted molecular fingerprints can then be used to help rank candidate metabolite IDs obtained based on predicted formula or measured precursor m/z of the unknown metabolite. This approach is particularly useful to help annotate metabolites whose corresponding MS/MS spectra cannot be matched with those in spectral libraries. We previously reported application of a convolutional neural network (CNN) for molecular fingerprint prediction using MS/MS spectra obtained from the MoNA repository and NIST 20. In this paper, we investigate high-dimensional representation of the spectral data and molecular fingerprints to improve accuracy in molecular fingerprint prediction.


Effect of duration of hemostatic compression on radial artery occlusion after transradial access.
To evaluate the effect of duration of hemostatic compression on the incidence of radial artery occlusion (RAO) after transradial coronary intervention. RAO occurs in 2-10% of patients after transradial access. The effect of duration of hemostatic compression on occurrence of RAO is unknown. Four hundred consecutive patients undergoing transradial coronary intervention were retrospectively analyzed. Group I (n = 200) patients received hemostatic compression for 6 hr after completion of the procedure, and group II (n = 200) patients for 2 hr after completion of the procedure. TR band was used for hemostasis. Demographic and procedural variables as well as early (24 hr) and chronic (30 days) RAO as well as bleeding events were recorded. Demographic as well as procedural variables were similar between group I and group II. Early radial artery occlusion (ERAO) occurred in 12% of patients in group I and 5.5% of patients in group II, the difference was statistically significant (P = 0.025). Chronic radial artery occlusion (CRAO) occurred in 8.5% of patients in group I and 3.5% of patients in group II, the difference was statistically significant (P = 0.035). Occlusive compression was the only independent predictor of radial artery occlusion [Odds ratio = 13.1, P = 0.001]. Two patients in group I and one in group II developed small hematoma (P = 0.161). Shorter duration of hemostatic compression is associated with a lower incidence of early and chronic RAO, without increase in bleeding complications, although maintaining radial patency during hemostatic compression, eliminates the adverse effect of duration of compression.


Hydroxylated metabolies of R,S-1-(tetrahydro-2-furanyl)-5-fluorouracil (Ftorafur) in rats and rabbits.
Two hydroxylated metabolies (M1 and M2) have been isolated from rabbit urine after administration of Ftorafur (FT). The structures of 3'-OH-FT and 4'-OH-FT were assigned to M1 and M2, respectively. A reverse-phase high performance liquid chromatography assay was developed for jeasuring FT, M1, M2, and 5-fluorouracil (FU) plasma levels. M1, M2, and FU were present in rabbit and rat plasma in greatly varying concentrations after FT administration. Pharmacokinetic studies suggest that FU formation proceeds via metabolic intermediate(s) and that the extent of FT activation is variable. A horse liver thymidine phosphorylase ,reparation capable of catalyzing the conversion of beta-ribo-2'-deoxy-5-fluorouracil to FU was inactive against FT and M1. However, 20% of M2 was converted to FU by this enzume, which suggests that the urinary metabolite M2 consisted of a mixture of enantiomers with 20% present in the natural beta-D configuration. The stereochemistry of M1 remains unknown. Hydroxylation of FT to beta-D-4'-OH-FT and subsequent cleavage to FU by thymidine phosphorylase represents one possible activation mechanism of FT to FU. ,owever, lack of correlation between plasma levels of M2 and FU indicates that this mode of metabolic activation may account for only part of the overall activation of FT in vivo.


Integrated molecular diagnosis of Theileria and Babesia species of cattle in Italy.
A reverse line blot hybridization (RLB) test was developed to specifically identify six Theileria spp. (T. annulata, T. parva, T. mutans, T. velifera, T. taurotragi, and T. buffeli/orientalis) and three Babesia spp. (B. bovis, B. bigemina, and B. divergens). No cross reaction was observed with other livestock pathogens (such as Anaplasma marginale, A. centrale, A. ovis, Cowdria ruminantium, Trypanosoma brucei, T. congolense, and T. vivax). This method was used to test bovine blood samples collected in Sicily in April and November, 1998. Preliminary results indicated that T. annulata and T. buffeli/orientalis were the main species observed in cattle blood. Babesia species represented 1.8% and 23.5% in April and November, respectively.


Neuropsychological impairment from acute low-level exposure to carbon monoxide.
To investigate the effects of acute low-level exposure to carbon monoxide on higher cognitive functions in healthy humans. An empirical study of the effects of low-level exposure to carbon monoxide on higher cognitive functions in young healthy volunteers and a matched nonexposed control group. A dormitory at the Hebrew University campus in Jerusalem, Israel. Forty-five student volunteers who were exposed to carbon monoxide from residential kerosene stoves for 1.5 to 2.5 hours (air carbon monoxide concentrations of 17-100 ppm; mean +/- SD, 61 +/- 24 ppm) served as the experimental group and 47 nonexposed students served as the control group. A battery of neuropsychological tests was administered to each participant including digit span, the revised Wechsler Memory Scale for verbal and figural memory, Trail-Making Test parts A and B, digit symbol, block design, and the Rey Auditory Verbal Learning Test. Venous blood carboxyhemoglobin (Hbco) levels in participants of the study group ranged from 0.01 to 0.11 (mean +/- SD, 0.04 +/- 0.03) and correlated with air carbon monoxide concentrations (r = 0.39; P = .01). The experimental group scored significantly lower than controls on the following tests: digit span forward (P = .02), short-term (P = .008) and long-term semantic memory (P = .008), digit symbol (P = .004), block design (P = .009), recall of figural memory (P = .02), and Trail-Making part A (P = .04). No significant differences were found between the experimental and control groups in other tests. The lower scores on neuropsychological tests indicate dysfunctions in memory, new learning ability, attention and concentration, tracking skills, visuomotor skills, abstract thinking, and visuospatial planing and processing. These dysfunctions correspond with previous reports of carbon monoxide neurotoxic effects in patients with moderate carbon monoxide poisoning. Low-level exposure to carbon monoxide results in impairment of higher cognitive functions. Neuropsychological testing appears to be sensitive in the detection of subtle neurologic dysfunctions resulting from carbon monoxide poisoning.


Patient-reported outcomes with durvalumab, with or without tremelimumab, plus chemotherapy as first-line treatment for metastatic non-small-cell lung cancer (POSEIDON).
In the phase 3 POSEIDON study, first-line tremelimumab plus durvalumab and chemotherapy significantly improved overall survival and progression-free survival versus chemotherapy in metastatic non-small-cell lung cancer (NSCLC). We present patient-reported outcomes (PROs). Treatment-naïve patients were randomized 1:1:1 to tremelimumab plus durvalumab and chemotherapy, durvalumab plus chemotherapy, or chemotherapy. PROs (prespecified secondary endpoints) were assessed using the European Organisation for Research and Treatment of Cancer 30-item core quality of life questionnaire version 3 (QLQ-C30) and its 13-item lung cancer module (QLQ-LC13). We analyzed time to deterioration (TTD) of symptoms, functioning, and global health status/quality of life (QoL) from randomization by log-rank test and improvement rates by logistic regression. 972/1013 (96 %) patients randomized completed baseline QLQ-C30 and QLQ-LC13 questionnaires, with scores comparable between treatment arms. Patients receiving tremelimumab plus durvalumab and chemotherapy versus chemotherapy had longer median TTD for all PRO items. Hazard ratios for TTD favored tremelimumab plus durvalumab and chemotherapy for all items except diarrhea; 95 % confidence intervals did not cross 1.0 for global health status/QoL, physical functioning, cognitive functioning, pain, nausea/vomiting, insomnia, constipation, hemoptysis, dyspnea, and pain in other parts. For durvalumab plus chemotherapy, median TTD was longer versus chemotherapy for all items except nausea/vomiting and diarrhea. Hazard ratios favored durvalumab plus chemotherapy for all items except appetite loss; 95 % confidence intervals did not cross 1.0 for global health status/QoL, physical functioning, role functioning, dyspnea, and pain in other parts. For both immunotherapy plus chemotherapy arms, improvement rates in all PRO items were numerically higher versus chemotherapy, with odds ratios > 1. Tremelimumab plus durvalumab and chemotherapy delayed deterioration in symptoms, functioning, and global health status/QoL compared with chemotherapy. Together with significant improvements in survival, these results support tremelimumab plus durvalumab and chemotherapy as a first-line treatment option in metastatic NSCLC.


Isolation and Identification of Flavones Responsible for the Antibacterial Activities of Tillandsia bergeri Extracts.
Plants are an everlasting inspiration source of biologically active compounds. Among these medicinal plants, the biological activity of extracts from some species of the Tillandsia genus has been studied, but the phytochemistry of the hardy species Tillandsia bergeri remains unknown. The aim of the present study was to perform the first phytochemical study of T. bergeri and to identify the compounds responsible for the antibacterial activity of T. bergeri extracts. Soxhlet extraction of predried and grinded leaves was first performed using four increasing polarity solvents. A bio-guided fractionation was performed using agar overlay bioautography as a screening method against 12 Gram-positive, Gram-negative, sensitive, and resistant bacterial strains. The results showed the inhibition of Gram-positive methicillin-sensitive Staphylococcus aureus ATCC 29213 (MSSA), methicillin-resistant S. aureus N-SARM-1 (MRSA), and Staphylococcus caprae ATCC 35538 by the dichloromethane fraction. A phytochemical investigation led to the isolation and identification by high-resolution mass spectrometry and nuclear magnetic resonance of the two flavones penduletin and viscosine, responsible for this antibacterial activity. For viscosine, the minimum inhibitory concentration (MIC) value is equal to 128 μg/mL against MSSA and is equal to 256 μg/mL against MRSA and S. caprae. The combination of these compounds with vancomycin and cloxacillin showed a decrease in MICs of the antibiotics. Penduletin showed synergistic activity when combined with vancomycin against MSSA (FICI < 0.258) and S. caprae (FICI < 0.5). Thus, unexplored Tillandsia species may represent a valuable source for potential antibiotics and adjuvants.


Is coulomb explosion a damaging mechanism for (125)IUdR?
To test the integrity of the thymine molecule that experiences an increasing number of charges due to the loss of Auger electrons emitted by the decay of incorporated 125I. Besides the radiation action of these electrons, Coulomb explosion is suspected to be an additional mechanism responsible for the strong radiotoxic effect of decaying DNA-incorporated 125I. The two-step decay process initiates a first Auger cascade within 10(-16) to 10(-14) s resulting in the release of about 7 electrons on average and a corresponding large positive charge on the 125Te daughter atom. Being part of iododeoxyuridine (125IUdR), the analogue of the DNA base thymine, the base is suddenly confronted with this charge. Experimentally, the situation was investigated with small molecules (CH3(125)I and C2H5(125)I) resulting in ion fragmentation in agreement with a Coulomb explosion model (Carlson and White, 1963, 1966). Semi-empirical quantum mechanical calculations on the Parametric Method 3 (PM3) level (Stewart, 1989a, 1989b) were performed and geometry optimisation was applied for the identification of stable molecule conformations. Subsequently, semiempirical molecular dynamics simulations allowed changes in the conformations to be studied as a function of time. First results show that there is no stable molecular configuration with a total charge of > or = +5e. PM3 calculations will not converge for such a charge located at the 125I/125Te position. This finding is supported by total energy considerations, which begin to favour a system of isolated atoms versus molecular bound atoms when the molecular charge is greater than +4e. The distribution of the partial charges indicates that most of the charge will remain on the tellurium atom with slight increases of charge at the other molecular partners within 125IUdR. Moreover, the molecular dynamics simulations reveal a breaking of chemical bonds between those atoms with the strongest charge increase. Coulomb explosion must be taken into account as a possible damaging mechanism following the decay of DNA-incorporated Auger electron emitters. Lobachevsky and Martin (2000) have identified the same mechanism to be responsible for part of strand breakage in oligo-deoxynucleotides. To elucidate a possible link between both damage patterns the molecular mechanics simulations have to be extended to larger parts of the DNA molecule.


Knowledge, Attitude, and Practice Towards Face-Mask Use Amid the Coronavirus 2019 Pandemic Among Public Transport Drivers in Gondar Town, A Cross-Sectional Study.
Background. Coronavirus disease (COVID-19) is a highly communicable virus that continues to interrupt livelihoods, predominantly those of low-income segments of society. For the prevention of respiratory infections like the current COVID-19 outbreak, face masks are considered an effective approach.Objective. This study intended to assess the knowledge, attitude, and practice of public transport drivers towards face mask use amid the COVID-19 pandemic in Gondar, Ethiopia.Methods. A cross-sectional study was conducted among 412 public transport drivers in Gondar town from July to September 2021. The study subjects were recruited using a simple random sampling technique after proportionally allocating the sample size from the total number of public transport drivers, and finally, study subjects were selected using the convenience sampling method to select the participant drivers working in Gondar Town. The data were collected by face-to-face interview administered questioners and an on-the-spot observational checklist.Results. A total of 412 public transport drivers have participated in the study. The mean age of the respondents was 32.75 years (±8.75 years). The majority of the participants were Bajaj drivers 193 (46.84%). Among the responders, 114 (27.67%) of them use radio to gather information about the pandemic and 50 (12.14%) of them had reported being caught by COVID-19. Meanwhile, only 32 (7.77%) were vaccinated against COVID-19. Drivers that had a diploma level of education were found to be 87.7% less knowledgeable than degree holders (AOR .123, 95% CI = .026, .573). Respondents that had good knowledge about COVID-19 and face masks were found to have 1.7 times more positive attitudes than those that had poor knowledge (AOR = 1.728, 95% CI = 1.150, 2.596). drivers whose family members have ever been caught by COVID-19 were found 2 times more likely to use face masks whenever they are working/driving (AOR = 2.173, 95% CI = 1.015, 4.652) than their counterparts.Conclusion. This study revealed a very low Knowledge, attitude, and practice of face mask use among public transport drivers in Gondar town. Periodic reinforcement and training programs are needed for all public transport drivers in each level for proper understanding and adherence to COVID-19 prevention protocols and the use of face masks.


Facile formation of a microporous chitosan hydrogel based on self-crosslinking.
A facile approach for the formation of microporous (chitosan) hydrogel scaffolds based on self-crosslinking is presented. It is simple and does not require any sacrificial porogen, toxic initiator/catalyst, harmful irradiation, or sophisticated equipment. The pore size, porosity, and mechanical properties of our hydrogels can be readily tuned.


[Iron deficiency and anemia in pregnant women from Chaco, Argentina].
Aiming at their prevention, to determine the prevalence of gestational iron deficiency and anemia, considering access to health care and associations with social and biological variables in the Province of el Chaco, Argentina. Three hundred and sixty four, randomly selected pregnant women from the whole province participated. Mean Hemoglobin (Hb) +/- SD dropped progressively from 118.4 +/- 11.0 to 112.1 +/- 11.5 g/L (p = 0.015) between the 1st and the 3rd gestational trimesters. Anemia prevalence (Hb < 110 g/L) was 17.4, 26.5 and 35.8%. From the 1st to the 3rd trimesters. Second trimester prevalence was 14.1% using Hb < 105 g/L as recently suggested. The ferritin geometric mean during the progressive gestational trimesters were 28.9; 16.0 and 11.1 micrograms/L. 21.7, 40.5 and 60.5% had ferritins < 12 micrograms/L, and 39.1, 58.9 and 83.2% had ferritins < 20 micrograms/L in these respective trimesters (p < 0.004 between trimesters in both cases). Both iron deficiency and anemia were significantly greater in women with unfinished primary education and with interpregnancy interval < 1 year. Seventy five percent of women had adequate number of antenatal visits but only 23% were taking iron supplements and 10% had stopped taking them. The prevention and correction of gestational iron deficiency and anemia must focus on the preconceptional period (inter-pregnancy spacing and increasing iron intake using all available means) as well as during pregnancy improving adherence to iron supplements by means of motivation of health workers and community.


Ca2+ and calpain mediate capsaicin-induced ablation of axonal terminals expressing transient receptor potential vanilloid 1.
Capsaicin is an ingredient in spicy peppers that produces burning pain by activating transient receptor potential vanilloid 1 (TRPV1), a Ca2+-permeable ion channel in nociceptors. Capsaicin has also been used as an analgesic, and its topical administration is approved for the treatment of certain pain conditions. The mechanisms underlying capsaicin-induced analgesia likely involve reversible ablation of nociceptor terminals. However, the mechanisms underlying these effects are not well understood. To visualize TRPV1-lineage axons, a genetically engineered mouse model was used in which a fluorophore is expressed under the TRPV1 promoter. Using a combination of these TRPV1-lineage reporter mice and primary afferent cultures, we monitored capsaicin-induced effects on afferent terminals in real time. We found that Ca2+ influx through TRPV1 is necessary for capsaicin-induced ablation of nociceptive terminals. Although capsaicin-induced mitochondrial Ca2+ uptake was TRPV1-dependent, dissipation of the mitochondrial membrane potential, inhibition of the mitochondrial transition permeability pore, and scavengers of reactive oxygen species did not attenuate capsaicin-induced ablation. In contrast, MDL28170, an inhibitor of the Ca2+-dependent protease calpain, diminished ablation. Furthermore, overexpression of calpastatin, an endogenous inhibitor of calpain, or knockdown of calpain 2 also decreased ablation. Quantitative assessment of TRPV1-lineage afferents in the epidermis of the hind paws of the reporter mice showed that EGTA and MDL28170 diminished capsaicin-induced ablation. Moreover, MDL28170 prevented capsaicin-induced thermal hypoalgesia. These results suggest that TRPV1/Ca2+/calpain-dependent signaling plays a dominant role in capsaicin-induced ablation of nociceptive terminals and further our understanding of the molecular mechanisms underlying the effects of capsaicin on nociceptors.


Clinical trial design and new therapies for pulmonary arterial hypertension.
Until 20 years ago the treatment of pulmonary arterial hypertension (PAH) was based on case reports and small series, and was largely ineffectual. As a deeper understanding of the pathogenesis and pathophysiology of PAH evolved over the subsequent two decades, coupled with epidemiological studies defining the clinical and demographic characteristics of the condition, a renewed interest in treatment development emerged through collaborations between international experts, industry and regulatory agencies. These efforts led to the performance of robust, high-quality clinical trials of novel therapies that targeted putative pathogenic pathways, leading to the approval of more than 10 novel therapies that have beneficially impacted both the quality and duration of life. However, our understanding of PAH remains incomplete and there is no cure. Accordingly, efforts are now focused on identifying novel pathogenic pathways that may be targeted, and applying more rigorous clinical trial designs to better define the efficacy of these new potential treatments and their role in the management scheme. This article, prepared by a Task Force comprised of expert clinicians, trialists and regulators, summarises the current state of the art, and provides insight into the opportunities and challenges for identifying and assessing the efficacy and safety of new treatments for this challenging condition.


A mathematical model for Neanderthal extinction.
A simple mathematical homogeneous model of competition is used to describe Neanderthal extinction in Europe. It considers two interacting species, Neanderthals and Early Modern Men, in the same ecological niche. Using paleontological data we claim that the parameter of similarity, between both species, fluctuates between 0.992 and 0.997. An extension of the model including migration (diffusion) is also discussed; nevertheless, extinction of Neanderthal seems unavoidable. Numerical analysis of travelling wave solutions (fronts) confirms the extinction. The wave-front-velocity is estimated from linear analysis and numerical simulations confirm this estimation. We conjecture a mathematical formulation for the principle of exclusion between competitive interacting species (Gause).


[Effect of pyruvate dehydrogenase coenzymes and mitochondrial proteins on the accumulation of [35S]lipoic acid].
Coenzymes introduced in the ratio, peculiar for pyruvate dehydrogenase complex into the medium containing fresh-isolated mitochondria and oxidation substrate--pyruvate increase accumulation of [35S] lipoate by these organelles. This process is highly stimulated by introducing either the only CoA or a coenzyme mixture (CoA, thiamine pyrophosphate, FAD, NAD). Addition of phosphate-extracted components of mitochondria and their protein fraction with coenzymes in the ratio indicated above provides maximum accumulation of [35S] lipoate by liver mitochondria. An equimolar mixture of coenzymes as well as protein components evoke no reliable variations in [35S] lipoate accumulation by albino rat liver mitochondria, while addition of the only thiamine pyrophosphate decreases this accumulation. Reconstruction of multienzyme complexes of coenzymes and apoenzymes on mitochondrion membranes accounts for the results obtained.


Association of Physician Orders for Life-Sustaining Treatment Form Use With End-of-Life Care Quality Metrics in Patients With Cancer.
Many patients with advanced cancer receive aggressive treatment near the end of life. The Physician Orders for Life-Sustaining Treatment (POLST) form is an innovation that converts patient preferences into medical orders. We compared the end-of-life care of patients with advanced cancer who had completed POLST forms with that of patients who had advance directives (ADs). We studied 2,159 West Virginians with ADs and/or POLST forms in the West Virginia e-Directive Registry who died as a result of cancer between January 2011 and February 2016. Data from the Vital Registration Office (date, site, and cause of death) were merged with those from the registry (form type, completion date, and hospice admission). The primary outcome variables were out-of-hospital death (OHD) and hospice admission. The percentage of patients with OHD was 85.7% for patients with POLST forms compared with 72.0% for those with ADs ( P < .001). The odds of OHD for patients with POLST forms were 2.33 times those of patients with ADs. The percentage of patients admitted to hospice was 49.9% for those with POLST forms compared with 27.0% for those with ADs ( P < .001). The odds of being admitted to hospice for patients with POLST forms were 2.69 times those of patients with ADs. Advance care planning with completion of POLST forms compared with ADs in patients with advanced cancer was associated with two quality end-of-life care metrics: OHD and hospice admission. Our study suggests that goals-of-care discussions including POLST form completion may improve end-of-life care for patients with advanced cancer.


Heparin mimetic peptide nanofibers promote angiogenesis.
New blood vessel formation (angiogenesis) is one of the most important processes required for functional tissue formation. Induction of angiogenesis is usually triggered by growth factors released by cells. Glycosaminoglycans (e.g., heparan sulphates) in the extracellular matrix aid in proper functioning of these growth factors. Therefore, exogeneous heparin or growth factors were required for promoting angiogenesis in previous regenerative medicine studies. Here we report for the first time induction of angiogenesis by a synthetic nanofibrous peptide scaffold without the addition of any exogenous growth factors or heparin. We designed and synthesized a self-assembling peptide amphiphile molecule that is functionalized with biologically active groups to mimic heparin. Like heparin, this molecule has the ability to interact with growth factors and effectively enhance their bioactivity. The nanofibers formed by these molecules were shown to form a 3D network mimicking the structural proteins in the extracellular matrix. Because of heparin mimicking capabilities of the peptide nanofibers, angiogenesis was induced without the addition of exogenous growth factors in vitro. Bioactive interactions between the nanofibers and the growth factors enabled robust vascularization in vivo as well. Heparin mimetic peptide nanofibers presented here provide new opportunities for angiogenesis and tissue regeneration by avoiding the use of heparin and exogenous growth factors. The synthetic peptide nanofiber scaffolds enriched with proper chemical functional groups shown in this study can be used to induce various desired physiological responses for tissue regeneration.


Longitudinal links between childhood peer victimization, internalizing and externalizing problems, and academic functioning: developmental cascades.
Developmental cascade models linking childhood peer victimization, internalizing and externalizing problems, and academic functioning were examined in a sample of 695 children assessed in Grade 3 (academic only) and Grades 5, 6, 7, and 8. Results revealed several complex patterns of associations in which poorer functioning in one domain influenced poorer outcomes in other areas. For example, a symptom driven pathway was consistently found with internalizing problems predicting future peer victimization. Support for an academic incompetence model was also found-- lower GPA in Grade 5, 6, and 7 was associated with more externalizing issues in the following year, and poor writing performance in Grade 3 predicted lower grades in Grade 5, which in turn predicted more externalizing problems in Grade 6. Results highlight the need to examine bidirectional influences and multifarious transactions that exist between peer victimization, mental health, and academic functioning over time.


Torn Between Valences: Mixed Emotions Predict Poorer Psychological Well-Being and Job Burnout.
Conflicting perspectives on whether mixed emotions are adaptive states which promote integrative processes or uncomfortable states which are agonizing make divergent predictions on whether mixed emotions in daily life are conducive towards psychological well-being. We examined this across three studies. Study 1 (N = 335, 135 males; M age = 41.31) examined associations between state mixed emotions and psychological well-being. Study 2 (N = 389; 160 males; M age = 40.86) examined associations between workplace-specific mixed emotions and workplace ill-being assessed based on burnout. Both direct measures and indirect measures were examined in Study 1 and 2, allowing some comparison of methodological issues related to their predictive validity. Study 3 (N = 3444; 1587 males; M age = 55.34) examined bidirectional relationships between an index of emotional ambivalence and psychological well-being about ten years later, controlling for baselines. Demographic covariates, positive emotions, and negative emotions were controlled for in all studies. Analyses were performed using latent variable structural equation modelling. After adjusting for all covariates, the direct measures of mixed emotions predicted poorer psychological well-being (Study 1) and greater burnout (Study 2). However, indirect measures were found to suffer from relatively greater multicollinearity and poorer predictive validity upon controlling for positive and negative emotions. In Study 3, emotional ambivalence predicted poorer long-term psychological well-being, while psychological well-being also predicted lower emotional ambivalence, supporting bidrectionality. The findings provide preliminary evidence that naturalistically experienced mixed emotions may be conflicting and unpleasant, with potentially negative implications for psychological well-being over and above positive and negative emotions. The online version contains supplementary material available at 10.1007/s10902-021-00493-z.


Joint immobilization induced hypoxic and inflammatory conditions in rat knee joints.
The purpose of this study was to examine the hypoxic and inflammatory conditions after immobilization in the joint capsule of rat knees. The unilateral knee joints of adult male rats were immobilized with an internal fixator (Im group) for 1 day, 3 days, and 1, 2, 4, 8, and 16 weeks. Sham-operated animals had holes drilled in the femur and tibia and screws inserted without a plate (control group). The number of cells and blood vessels in the capsule were histologically examined. The hypoxic condition in the capsule was histologically examined with a Hypoxyprobe™-1. The gene expressions related to the hypoxic (hypoxia inducible factor-1α, vascular endothelial growth factor, and fibroblast growth factor 2) and inflammatory conditions [interleukin-6 (IL-6), IL-1α, IL-1β, tumor necrosis factor-α, and tumor necrosis factor-β] were evaluated by quantitative reverse transcription polymerase chain reaction. The number of cells was unchanged at 1 day in the two groups; however, the number significantly increased at 3 days in the Im group. The number of blood vessels in the Im group gradually decreased. Strong immunostaining of Hypoxyprobe™-1 around the blood vessels was observed in the Im group. The gene expressions of hypoxia inducible factor-1α and fibroblast growth factor 2 were significantly higher in the Im group compared with those in the control group. The gene expressions of IL-6, IL-1α, IL-1β, and tumor necrosis factor-β were significantly higher in the Im group compared with those in the control group. These data indicated that joint immobilization induced hypoxic and inflammatory conditions in the joint capsule, which might be an initiating factor for joint contracture.


Ganetespib induces G2/M cell cycle arrest and apoptosis in gastric cancer cells through targeting of receptor tyrosine kinase signaling.
Heat shock protein 90 (HSP90) regulates several important cellular processes via its repertoire of 'client proteins'. These client proteins have been found to play fundamental roles in signal transduction, cell proliferation, cell cycle progression and survival, as well as other features of malignant cells, such as invasion, tumor angiogenesis and metastasis. Thus, HSP90 is an emerging target for cancer therapy. To this end, we evaluated ganetespib (STA-9090), a novel and potent HSP90 inhibitor, for its activity in gastric cancer cell lines. Ganetespib significantly inhibited the proliferation of AGS and N87 human gastric cancer cell lines and potently induced G2/M cell cycle arrest and apoptosis. Upregulation of cleaved poly(ADP-ribose) polymerase (c-PARP), c-caspase-3, c-caspase-8 and c-caspase-9 and suppression of gastric cancer‑associated HSP90 client proteins, including ErbB2, Erk, Akt, mTOR, GSK3 and Src, were observed in ganetespib-treated cells. These findings demonstrate that the ganetespib-induced mechanism of cell growth inhibition involves the activation of death receptor and mitochondrial pathways and the inhibition of receptor tyrosine kinase signaling pathways. Our study implicates ganetespib as a potential strategy for gastric cancer treatment, which warrants further preclinical and clinical investigation.


Comparison of healing rate in diabetes-related foot ulcers with low frequency ultrasonic debridement versus non-surgical sharps debridement: a randomised trial protocol.
Foot ulceration has been reported as the leading cause of hospital admission and amputation in individuals with diabetes. Diabetes-related foot ulcers require multidisciplinary management and best practice care, including debridement, offloading, dressings, management of infection, modified footwear and management of extrinsic factors.Ulcer debridement is a commonly applied management approach involving removal of non-viable tissue from the ulcer bed. Different methods of debridement have been reported in the literature including autolytic debridement via moist wound healing, mechanical debridement utilising wet to dry dressings, theatre based sharps debridement, biological debridement, non-surgical sharps debridement and newer technology such as low frequency ultrasonic debridement. People with diabetes and a foot ulcer, referred to and treated by the Podiatry Department at Monash Health and who meet the inclusion criteria will be invited to participate in this randomised controlled trial. Participants will be randomly and equally allocated to either the non-surgical sharps debridement (control) or low frequency ultrasonic debridement (intervention) group (n = 322 ulcers/n = 108 participants).Where participants have more than one ulcer, only the participant will be randomised, not the ulcer. An investigator not involved in participant recruitment or assessment will be responsible for preparing the random allocation sequence and envelopes.Each participant will receive weekly treatment for six months including best practice podiatric management. Each ulcer will be measured on a weekly basis by calculating total area in centimetres squared. Measurement will be undertaken by a trained research assistant to ensure outcomes are blinded from the treating podiatrist. Another member of the research team will assess the final primary outcome. The primary aim of this study is to compare healing rates for diabetes-related foot ulcers using non-surgical sharps debridement versus low frequency ultrasonic debridement over a six month period. The primary outcome measure for this study is the proportion of ulcers healed by the six month follow-up period.Secondary outcomes will include a quality of life measure, assessment of pain and health care resource use between the two treatment modalities. Australian New Zealand Clinical Trial Registry: ACTRN12612000490875.


INTEREST: INteractive Tool for Exploring REsults from Simulation sTudies.
Simulation studies allow us to explore the properties of statistical methods. They provide a powerful tool with a multiplicity of aims; among others: evaluating and comparing new or existing statistical methods, assessing violations of modelling assumptions, helping with the understanding of statistical concepts, and supporting the design of clinical trials. The increased availability of powerful computational tools and usable software has contributed to the rise of simulation studies in the current literature. However, simulation studies involve increasingly complex designs, making it difficult to provide all relevant results clearly. Dissemination of results plays a focal role in simulation studies: it can drive applied analysts to use methods that have been shown to perform well in their settings, guide researchers to develop new methods in a promising direction, and provide insights into less established methods. It is crucial that we can digest relevant results of simulation studies. Therefore, we developed INTEREST: an INteractive Tool for Exploring REsults from Simulation sTudies. The tool has been developed using the Shiny framework in R and is available as a web app or as a standalone package. It requires uploading a tidy format dataset with the results of a simulation study in R, Stata, SAS, SPSS, or comma-separated format. A variety of performance measures are estimated automatically along with Monte Carlo standard errors; results and performance summaries are displayed both in tabular and graphical fashion, with a wide variety of available plots. Consequently, the reader can focus on simulation parameters and estimands of most interest. In conclusion, INTEREST can facilitate the investigation of results from simulation studies and supplement the reporting of results, allowing researchers to share detailed results from their simulations, readers to explore them freely.


Circulating Proteins as Diagnostic Markers in Gastric Cancer.
Gastric cancer (GC) is a highly malignant disease affecting humans worldwide and has a poor prognosis. Most GC cases are detected at advanced stages due to the cancer lacking early detectable symptoms. Therefore, there is great interest in improving early diagnosis by implementing targeted prevention strategies. Markers are necessary for early detection and to guide clinicians to the best personalized treatment. The current semi-invasive endoscopic methods to detect GC are invasive, costly, and time-consuming. Recent advances in proteomics technologies have enabled the screening of many samples and the detection of novel biomarkers and disease-related signature signaling networks. These biomarkers include circulating proteins from different fluids (e.g., plasma, serum, urine, and saliva) and extracellular vesicles. We review relevant published studies on circulating protein biomarkers in GC and detail their application as potential biomarkers for GC diagnosis. Identifying highly sensitive and highly specific diagnostic markers for GC may improve patient survival rates and contribute to advancing precision/personalized medicine.


Don't go there - It's not a nice place: Older adults' experiences of delirium.
Delirium is a common neuropsychiatric disorder that causes fluctuations in consciousness and attention, impairments in cognitive functioning and information processing, and changes in how individuals perceive what is going on around them. Delirium is associated with increased mortality, ongoing impairment in cognitive functioning, and a high possibility of discharge to residential care. The experience of delirium may be distressing for the patient and their family. Despite the frequency of delirium in hospitalized elderly patients, there is a dearth of literature that examines their experience of this phenomenon, and how it affects individuals as they continue their lives. This study uses descriptive qualitative methodology to explore the question: 'What is the experience of delirium for older adults during hospitalisation?' Data were collected from older adults who had received hospital care in a tertiary general hospital setting. Seven participants were recruited between January and June 2017. Semi-structured individual interviews were used to gather data which was analysed using content analysis. Four themes were identified. These were sense of confusion, disrupted sense of autonomy, perceptual disturbances, and emotional response. Participants exercised agency in the way they responded to these experiences. The study highlighted the need for delirium prevention, and education to improve nurses' recognition, understanding, and management of delirium. In particular, there is a need for nurses to attend to the psychological and emotional experience of delirium.


Electrostimulation as a prosthesis for repair of information flow in a computer model of neocortex.
Damage to a cortical area reduces not only information transmitted to other cortical areas, but also activation of these areas. This phenomenon, whereby the dynamics of a follower area are dramatically altered, is typically manifested as a marked reduction in activity. Ideally, neuroprosthetic stimulation would replace both information and activation. However, replacement of activation alone may be valuable as a means of restoring dynamics and information processing of other signals in this multiplexing system. We used neuroprosthetic stimulation in a computer model of the cortex to repair activation dynamics, using a simple repetitive stimulation to replace the more complex, naturalistic stimulation that had been removed. We found that we were able to restore activity in terms of neuronal firing rates. Additionally, we were able to restore information processing, measured as a restoration of causality between an experimentally recorded signal fed into the in silico brain and a cortical output. These results indicate that even simple neuroprosthetics that do not restore lost information may nonetheless be effective in improving the functionality of surrounding areas of cortex.


Fetal inflammation associated with minimal acute morbidity in moderate/late preterm infants.
To determine whether exposure to acute chorioamnionitis and fetal inflammation caused short-term adverse outcomes. This is a prospective observational study: subjects were mothers delivering at 32-36 weeks gestation and their preterm infants at a large urban tertiary level III perinatal unit (N=477 infants). Placentae and fetal membranes were scored for acute histological chorioamnionitis based on the Redline criteria. Fetal inflammation was characterised by histological diagnosis of funisitis (umbilical cord inflammation), increased cord blood cytokines measured by ELISA, and activation of the inflammatory cells infiltrating the placenta and fetal membranes measured by immunohistology. Maternal and infant data were collected. Twenty-four per cent of 32-36-week infants were exposed to histological chorioamnionitis and 6.9% had funisitis. Immunostaining for leucocyte subsets showed selective infiltration of the placenta and fetal membranes with activated neutrophils and macrophages with chorioamnionitis. Interleukin (IL) 6, IL-8 and granulocyte colony-stimulating factor were selectively increased in the cord blood of preterm infants with funisitis. Compared with infants without chorioamnionitis, funisitis was associated with increased ventilation support during resuscitation (43.8% vs 15.4%) and more respiratory distress syndrome postnatally (27.3% vs 10.2%) in univariate analysis. However, these associations disappeared after adjusting for prematurity. Despite fetal exposure to funisitis, increased cord blood cytokines and activated placental inflammatory cells, we could not demonstrate neonatal morbidity specifically attributable to fetal inflammation after adjusting for gestational age in moderate and late preterm infants.


An Umbrella Review of Self-Management Interventions for Health Conditions With Symptom Overlap With Traumatic Brain Injury.
To synthesize evidence for the effectiveness of self-management interventions for chronic health conditions that have symptom overlap with traumatic brain injury (TBI) in order to extract recommendations for self-management intervention in persons with TBI. An umbrella review of existing systematic reviews and/or meta-analyses of randomized controlled trials or nonrandomized studies targeting self-management of chronic conditions and specific outcomes relevant to persons with TBI. A comprehensive literature search of 5 databases was conducted using PRISMA guidelines. Two independent reviewers conducted screening and data extraction using the Covidence web-based review platform. Quality assessment was conducted using criteria adapted from the Assessing the Methodological Quality of Systematic Reviews-2 (AMSTAR-2). A total of 26 reviews met the inclusion criteria, covering a range of chronic conditions and a range of outcomes. Seven reviews were of moderate or high quality and focused on self-management in persons with stroke, chronic pain, and psychiatric disorders with psychotic features. Self-management interventions were found to have positive effects on quality of life, self-efficacy, hope, reduction of disability, pain, relapse and rehospitalization rates, psychiatric symptoms, and occupational and social functioning. Findings are encouraging with regard to the effectiveness of self-management interventions in patients with symptoms similar to those of TBI. However, reviews did not address adaptation of self-management interventions for those with cognitive deficits or for populations with greater vulnerabilities, such as low education and older adults. Adaptations for TBI and its intersection with these special groups may be needed.


CUG repeat binding protein (CUGBP1) interacts with the 5' region of C/EBPbeta mRNA and regulates translation of C/EBPbeta isoforms.
The transcription factor CCAAT/enhancer binding protein beta, C/EBPbeta, plays a significant role in the regulation of hepatocyte growth and differentiation. A single mRNA coding for C/EBPbeta produces several protein isoforms. Two pathways for generation of low molecular weight C/EBPbeta isoforms have been described: specific proteolytic cleavage and initiation of translation from different AUG codons of C/EBPbeta mRNA. A truncated C/EBPbeta isoform, LIP, is induced in rat livers in response to partial hepatectomy (PH) via the alternative translation mechanism. Here we present evidence that CUG repeat binding protein, CUGBP1, interacts with the 5' region of C/EBPbeta mRNA and regulates translation of C/EBPbeta isoforms. Two binding sites for CUGBP1 are located side by side between the first and second AUG codons of C/EBPbeta mRNA. One binding site is observed in an out of frame short open reading frame (sORF) that has been previously shown to regulate initiation of translation from different AUG codons of C/EBPbeta mRNA. Analysis of cytoplasmic and polysomal proteins from rat liver after PH showed that CUGBP1 is associated with polysomes that translate low molecular weight isoforms of C/EBPbeta. The binding activity of CUGBP1 to the 5' region of C/EBPbeta mRNA shows increased association with these polysomal fractions after PH. Addition of CUGBP1 into a cell-free translation system leads to increased translation of low molecular weight isoforms of C/EBPbeta. Our data demonstrate that CUGBP1 protein is an important component for the regulation of initiation from different AUG codons of C/EBPbeta mRNA.


Development of a hormone neutralizing vaccine, using GnRH-glycys-PPD, for use in the treatment of oestrogen-dependent disorders.
The aim of this study was to develop an effective and nontoxic vaccine, suitable for use in humans, which was capable of effectively controlling oestrogen levels. Female Sprague-Dawley rats were immunized with a conjugated analogue of gonadotrophin releasing hormone, GnRH-glycys-PPD. This resulted in high levels of neutralizing antibody which disrupted GnRH function and consequently caused a reduction in serum oestrogen. The effect of oestrogen deprivation correlated well with ovarian failure and gonadal atrophy. An examination was made of various adjuvants in conjunction with the analogue to determine the suitability of the combinations in the formulation of an effective human vaccine. This investigation included a novel adjuvant, non-ionic surfactant vesicles (NISV); the results showed that NISV are completely nontoxic and in terms of potentiating and sustaining an immune response, compare favourably with Freund's adjuvant and alum. In addition the long term effects of immunization were investigated and the data showed that immunoneutralization of GnRH effectively suppresses fertility on a long-term basis.


Immunomodulatory role of branched-chain amino acids.
Branched-chain amino acids (BCAAs) have been associated with immunomodulation since the mid-1970s and 1980s and have been used in the nutritional therapy of critically ill patients. Evidence shows that BCAAs can directly contribute to immune cell function, aiding recovery of an impaired immune system, as well as improving the nutritional status in cancer and liver diseases. Branched-chain amino acids may also play a role in treatment of patients with sepsis or trauma, contributing to improved clinical outcomes and survival. Branched-chain amino acids, especially leucine, are activators of the mammalian target of rapamycin (mTOR), which, in turn, interacts with several signaling pathways involved in biological mechanisms of insulin action, protein synthesis, mitochondrial biogenesis, inflammation, and lipid metabolism. Although many in vitro and human and animal model studies have provided evidence for the biological activity of BCAAs, findings have been conflicting, and the mechanisms of action of these amino acids are still poorly understood. This review addresses several aspects related to BCAAs, including their transport, oxidation, and mechanisms of action, as well as their role in nutritional therapy and immunomodulation.


Quantitative assessment of gastric atrophy using the syntactic structure analysis.
To assess the topographical relation between gastric glands, using the minimum spanning tree (MST), to derive both a model of neighbourhood and quantitative representation of the tissue's architecture, to assess the characteristic features of gastric atrophy, and to assess the grades of gastric atrophy. Haematoxylin and eosin stained sections from corporal and antral biopsy specimens (n = 139) from normal patients and from patients with nonatrophic gastritis and atrophic gastritis of grades 1, 2, and 3 (Sydney system) were assessed by image analysis system (Prodit 5.2) and 11 syntactic structure features were derived. These included both line and connectivity features. Syntactic structure analysis was correlated with the semiquantitative grading system of gastric atrophy. The study showed significant reductions in the number of points and the length of MST in both body and antrum. The standard deviation of the length of MST was significantly increased in all grades of atrophy. The connectivity to two glands was the highest and most affected by the increased grade of atrophy. The reciprocal values of the Wiener, Randic, and Balaban indices showed significant changes in the volume of gland, abnormality in the shape of glands, and changes in irregularity and branching of the glands in both types of gastric mucosa. There was a complete separation in the MST, connectivity, and index values between low grade and high grade gastric atrophy. (1) Gastric atrophy was characterised by loss of the gland, variation in the volume, reduction in the neighbourhood, irregularity in spacing, and abnormality in the shape of the glands. (2) Syntactic structure analysis significantly differentiated minor changes in gastric gland (low grade atrophy) from high grade atrophy of clinical significance. (3) Syntactic structure analysis is a simple, fast, and highly reproducible technique and appears a promising method for quantitative assessment of atrophy.


Sex Differences in Vascular Reactivity to Angiotensin II During the Evolution of Myocardial Infarction.
The influence of sex on the vascular response during the progression of myocardial infarction (MI) has not been extensively studied. In this work, we analyzed the differences of the vasoconstriction induced by angiotensin II (Ang II) in the absence and presence of valsartan (200 nM) on the aortic rings of male and female Wistar rats at 2, 4, 24, and 48 hours and 1, 2, 3, and 4 weeks after induction of MI. In the aortic rings of males, an increase was observed in the contractile response that lasts up to 4 weeks; on females, this effect is diminished since 48 hours until reaching sham group values at 2 weeks of coronary occlusion. The incubation of valsartan generated greater reduction on vasoconstriction in males than females. In relation to the determination of infarct areas, we found them between 30% and 40% in all experimental groups. In addition, the index of hypertrophy was determined and no significant changes were observed in female rats, while in males we reported an increase at 2, 3, and 4 weeks. In conclusion, we found differences in vascular reactivity due to sex, as well as on the response of Ang II via AT1 during the evolution of MI.


Clusterin/Apolipoprotein J immunolocalization on carotid artery is affected by TNF-alpha, cigarette smoking and anti-platelet treatment.
Clusterin (CLU) /Apolipoprotein J is a protein biosensor of oxidative stress and inflammation, which is upregulated in many pathological processes including atherosclerosis. Previous studies have shown that in aortic tissue, CLU expression increases with atherosclerotic lesion progression and it has been coupled with vascular damage and coronary artery disease. A few studies enter into CLU and carotid atherosclerosis while the apolipoprotein's expression on human carotid tissue and its association with parameters related to the disease development has not been examined. The present study was designed to reveal the relationships between the degree of CLU immunolocalization on carotid artery and demographic characteristics, blood parameters and pharmacological treatment of patients underwent internal carotid artery endarterectomy. CLU expression was detected by immunohistochemistry in 42 carotid endarterectomy specimens. Patients' serum levels of tumor necrosis factor-a (TNF-a), interleukin-6 (IL-6), high sensitive C-reactive protein (hsCRP) and classical parameters related to atherosclerosis such as lipid profile, as well as thrombosis related parameters such as fibrinogen, antithrombin III, protein C and protein S were determined. Demographic characteristics, smoking habits and the use of medications were recorded. Comparisons between groups were performed by students't-test and analysis of variance. Independent associations with CLU expression on carotid tissue were denoted by linear regression analysis. CLU imuunolocalization was denser in smokers than in non-smokers (p = 0.041) while it was rarefied in specimens of patients on cropidogrel treatment (p = 0.045) compared to the rest not taking this medication. Clopidogrel intake was independent predictor of lower CLU expression on carotid artery (p =0.045). CLU was positively correlated with serum TNF-a concentration (r = 0.33, p = 0.040) that was independent predictor of higher expression of the apolipoprotein (p = 0.001). IL-6, hsCRP and classical parameters related to atherosclerosis and thrombosis were not associated with CLU immunolocalization. Our study suggests that CLU expression on carotid artery is affected by TNF-alpha, cigarette smoking confirming its association with oxidative and cellular stress and anti-platelet medication reflecting the protective effects of such pharmacological treatment on vascular wall.


Neurotensin and acetylcholine evoke common responses in frog oocytes injected with rat brain messenger ribonucleic acid.
1. The intracellular reaction mechanism underlying electrophysiological responses evoked by neurotensin (NT) was studied using Xenopus laevis oocytes injected with poly (A)+ messenger ribonucleic acid (mRNA) isolated from rat brains. 2. A few days after the injection of mRNA, oocytes were found to acquire sensitivity to NT and substance P. 3. Under voltage-clamp conditions (-60 mV), application of NT to mRNA-injected oocytes produced transient and oscillatory inward currents which began after a delay of several tens of seconds. These inward currents were accompanied by an increase in membrane conductance. 4. NT receptors on mRNA-injected oocytes showed essentially the same pharmacological properties as those of native NT receptors. 5. The NT response showed desensitization and was not readily recovered even after extensive washing of cells for more than 30 min. 6. NT response was suppressed when the muscarinic acetylcholine (ACh) response of the same cell, which was also induced by the same mRNA, was desensitized by a large dose of ACh. 7. NT response and ACh response showed many similarities: they were both inhibited by pertussis toxin and intracellular ethyleneglycol-bis-(beta-aminoethylether) N, N'-tetraacetic acid (EGTA), mimicked by intracellularly injected inositol 1, 4, 5-trisphosphate (InsP3), and suppressed when cell response to InsP3 was desensitized by a large dose of InsP3. Reversal-potential analyses indicated that both responses were mediated by an increase in membrane permeability to Cl-. 8. It is concluded that NT responses and muscarinic ACh responses of Xenopus oocytes induced by rat brain mRNA may most likely share a common reaction mechanism. The reaction sequence includes the activation of receptors, activation of inhibitory guanine nucleotide-binding regulatory protein, production of InsP3, intracellular Ca2+ mobilization, and increased membrane permeability to Cl-.


Cinnamomum cassia bark in two herbal formulas increases life span in Caenorhabditis elegans via insulin signaling and stress response pathways.
Proving the efficacy and corresponding mode of action of herbal supplements is a difficult challenge for evidence-based herbal therapy. A major hurdle is the complexity of herbal preparations, many of which combine multiple herbs, particularly when the combination is assumed to be vitally important to the effectiveness of the herbal therapy. This issue may be addressed through the use of contemporary methodology and validated animal models. In this study, two commonly used traditional herbal formulas, Shi Quan Da Bu Tang (SQDB) and Huo Luo Xiao Ling Dan (HLXL) were evaluated using a survival assay and oxidative stress biomarkers in a well-established C. elegans model of aging. HLXL is an eleven herb formula modified from a top-selling traditional herbal formula for the treatment of arthritic joint pain. SQDB consists of ten herbs often used for fatigue and energy, particularly in the aged. We demonstrate here that SQDB significantly extend life span in a C. elegans model of aging. Among all individual herbs tested, two herbs Cinnamomum cassia bark (Chinese pharmaceutical name: Cinnamomi Cortex, CIN) and Panax ginseng root (Chinese pharmaceutical name: Ginseng Radix, GS) significantly extended life span in C. elegans. CIN in both SQDB and HLXL formula extended life span via modulation of multiple longevity assurance genes, including genes involved in insulin signaling and stress response pathways. All the life-span-extending herbs (SQDB, CIN and GS) also attenuated levels of H2O2 and enhanced small heat shock protein expression. Furthermore, the life span-extending herbs significantly delayed human amyloid beta (Abeta)-induced toxicity in transgenic C. elegans expressing human Abeta. These results validate an invertebrate model for rapid, systematic evaluation of commonly used Chinese herbal formulations and may provide insight for designing future evidence-based herbal therapy(s).


Inhibitory Metaplasticity in Juvenile Stressed Rats Restores Associative Memory in Adulthood by Regulating Epigenetic Complex G9a/GLP.
Exposure to juvenile stress was found to have long-term effects on the plasticity and quality of associative memory in adulthood, but the underlying mechanisms are still poorly understood. Three- to four week-old male Wistar rats were subjected to a 3-day juvenile stress paradigm. Their electrophysiological correlates of memory using the adult hippocampal slice were inspected to detect alterations in long-term potentiation and synaptic tagging and capture model of associativity. These cellular alterations were tied in with the behavioral outcome by subjecting the rats to a step-down inhibitory avoidance paradigm to measure strength in their memory. Given the role of epigenetic response in altering plasticity as a repercussion of juvenile stress, we aimed to chart out the possible epigenetic marker and its regulation in the long-term memory mechanisms using quantitative reverse transcription polymerase chain reaction. We demonstrate that even long after the elimination of actual stressors, an inhibitory metaplastic state is evident, which promotes synaptic competition over synaptic cooperation and decline in latency of associative memory in the behavioral paradigm despite the exposure to novelty. Mechanistically, juvenile stress led to a heightened expression of the epigenetic marker G9a/GLP complex, which is thus far ascribed to transcriptional silencing and goal-directed behavior. The blockade of the G9a/GLP complex was found to alleviate deficits in long-term plasticity and associative memory during the adulthood of animals exposed to juvenile stress. Our data provide insights on the long-term effects of juvenile stress that involve epigenetic mechanisms, which directly impact long-term plasticity, synaptic tagging and capture, and associative memory.


Implementing population-based mass drug administration for malaria: experience from a high transmission setting in North Eastern Uganda.
Mass drug administration (MDA) is a suggested mean to accelerate efforts towards elimination and attainment of malaria-free status. There is limited evidence of suitable methods of implementing MDA programme to achieve a high coverage and compliance in low-income countries. The objective of this paper is to assess the impact of this MDA delivery strategy while using coverage measured as effective population in the community and population available. Population-based MDA was implemented as a part of a larger program in a high transmission setting in Uganda. Four rounds of interventions were implemented over a period of 2 years at an interval of 6 to 8 months. A housing and population census was conducted to establish the eligible population. A team of 19 personnel conducted MDA at established village meeting points as distribution sites at every village. The first dose of dihydroartemisinin-piperaquine (DHA-PQ) was administered via a fixed site distribution strategy by directly observed treatment on site, the remaining doses were taken at home and a door-to-door follow up strategy was implemented by community health workers to monitor adherence to the second and third doses. Based on number of individuals who turned up at the distribution site, for each round of MDA, effective coverage was 80.1%, 81.2%, 80.0% and 80% for the 1st, 2nd, 3rd and 4th rounds respectively. However, coverage based on available population at the time of implementing MDA was 80.1%, 83.2%, 82.4% and 82.9% for rounds 1, 2, 3 and 4, respectively. Intense community mobilization using community structures and mass media facilitated community participation and adherence to MDA. A hybrid of fixed site distribution and door-to-door follow up strategy of MDA delivery achieved a high coverage and compliance and seemed feasible. This model can be considered in resource-limited settings.


Heterogeneity of the Fatty Acid Composition of Japanese Placentae for Determining the Perinatal Fatty Acid Status: a Methodological Study.
Analysis of the placenta can be a useful way to determine the fatty acid (FA) status of pregnant women and neonates since this large organ can be obtained easily and non-invasively. Although several studies have been conducted on using placental tissue for FA analysis, the sampling methods have not been thoroughly examined. The aim of the present study was to determine a suitable method of sampling human placentae by focusing on their heterogeneity. Twenty-four placentae were collected from healthy pregnant Japanese women in the Miyagi Prefecture of Japan. Five of them were used to compare the FA composition between the peripheral area and the central area of the placentae. The other 19 were examined to determine differences in the FA composition between the fetal and maternal side. Placental tissue crude lipid was saponified, and methyl was esterified with 14% sodium boron trifluoride methanol for gas chromatography with flame ionization detector analysis. Fifty-six peaks were detected from the methyl esters of the placental total lipid, and 33 of those were identified as FA methyl esters. There were considerable variations in the FA composition, as the variation was low in the central parts and high in the peripheral parts of the placentae. The 18:1n-9 and 18:2n-6 levels were higher in the fetal side, whereas the 20:3n-6, 20:4n-6, and 22:6n-3 levels were higher in the maternal side. These findings indicate the presence of heterogeneity in the FA composition of human placenta, and they suggest the necessity for standardizing the sampling method so that the placental tissue can be used to determine the FA status.


Therapeutic horticulture in clinical depression: a prospective study of active components.
This paper is a report of a study conducted to assess change in depression severity, perceived attentional capacity and rumination (brooding) in individuals with clinical depression during a therapeutic horticulture programme and to investigate if the changes were mediated by experiences of being away and fascination. Individuals with clinical depression suffer from distortion of attention and rumination. Interventions can help to disrupt maladaptive rumination and promote restoration of depleted attentional capacity. A single-group study was conducted with a convenience sample of 28 people with clinical depression in 2009. Data were collected before, twice during, and immediately after a 12-week therapeutic horticulture programme, and at 3-month follow-up. Assessment instruments were the Beck Depression Inventory, Attentional Function Index, Brooding Scale, and Being Away and Fascination subscales from the Perceived Restorativeness Scale. Mean Beck Depression Inventory scores declined by 4.5 points during the intervention (F = 5.49, P = 0.002). The decline was clinically relevant for 50% of participants. Attentional Function Index scores increased (F = 4.14, P = 0.009), while Brooding scores decreased (F = 4.51, P = 0.015). The changes in Beck Depression Inventory and Attentional Function Index scores were mediated by increases in Being Away and Fascination, and decline in Beck Depression Inventory scores was also mediated by decline in Brooding. Participants maintained their improvements in Beck Depression Inventory scores at 3-month follow-up. Being away and fascination appear to work as active components in a therapeutic horticulture intervention for clinical depression.


In vivo prooxidant state in Werner syndrome (WS): results from three WS patients and two WS heterozygotes.
The hypothesis was tested that Werner syndrome (WS) phenotype might be associated with an in vivo prooxidant state. A set of redox-related endpoints were measured in three WS patients, two of their parents, and 99 controls within a study of some cancer-prone and/or ageing-related genetic disorders. The following analytes were measured: (a) leukocyte 8-hydroxy-2'-deoxyguanosine; (b) glutathione from whole blood, and (c) plasma levels of glyoxal, methylglyoxal, 8-isoprostane, and some plasma antioxidants (uric acid, ascorbic acid, alpha- and gamma-tocopherol). Leukocyte 8-hydroxy-2'-deoxyguanosine levels showed a significant increase in the 3 WS patients vs. 85 controls (p<10(-7)). The disulfide glutathione:glutahione ratio was significantly altered in WS patients (p=0.005). Glyoxal and methylglyoxal levels were significantly increased (p=0.018 and p=0.007, respectively). The plasma levels of uric acid (p=0.002) and ascorbic acid (p=0.003) were also increased significantly in WS patients and in their parents. No significant alterations were found in the plasma levels of alpha- and gamma-tocopherol, nor of 8-isoprostane. This is the first report of in vivo alterations of oxidative stress parameters in WS patients. Further investigations on more extensive study populations are warranted to verify the relevance of an in vivo prooxidant state in WS patients.


Negotiations about helpfulness--the relationship between formal and informal care in home care arrangements.
Informal caregiving by family members is the most common way of caring for sick people at home. However, the number of care arrangements, in which both formal (nurses) and informal (family members) caregivers are involved, is considerable and increasing. Despite implicit assumptions in research that the involvement of nurses in home care arrangements is inherently beneficial, there is evidence that their involvement may have a destabilising effect. The purpose of this study was to investigate the relationship between nurses and family caregivers and its impact on the actual care that is provided. Eighty-eight interviews with family caregivers (n = 57) and nurses (n = 31) were conducted in Germany and analysed according to the Grounded Theory methodology. The relationship between formal and informal care is an encounter of two quite different perspectives that is focused on a negotiation process about caregiving work and the helpfulness of the actions taken and the interventions used. For family caregivers, it is determined by the goal of facilitating work and care for their sick family member. The nurses' work is characterised by a process of shaping different realities in different homes. The results reveal the processes that lead to the involvement of nurses into home care arrangements and offer a deeper understanding of the negotiation processes between formal and informal caregivers. To provide sufficient support in home care, nurses need the ability to engage in negotiation processes that take the whole home care arrangement into account. Developmental work is needed to design services that are helpful for family caregivers.


Agdc1p - a Gallic Acid Decarboxylase Involved in the Degradation of Tannic Acid in the Yeast Blastobotrys (Arxula) adeninivorans.
Tannins and hydroxylated aromatic acids, such as gallic acid (3,4,5-trihydroxybenzoic acid), are plant secondary metabolites which protect plants against herbivores and plant-associated microorganisms. Some microbes, such as the yeast Arxula adeninivorans are resistant to these antimicrobial substances and are able to use tannins and gallic acid as carbon sources. In this study, the Arxula gallic acid decarboxylase (Agdc1p) which degrades gallic acid to pyrogallol was characterized and its function in tannin catabolism analyzed. The enzyme has a higher affinity for gallic acid (Km -0.7 ± 0.2 mM, kcat -42.0 ± 8.2 s-1) than to protocatechuic acid (3,4-dihydroxybenzoic acid) (Km -3.2 ± 0.2 mM, kcat -44.0 ± 3.2 s-1). Other hydroxylated aromatic acids, such as 3-hydroxybenzoic acid, 4-hydroxybenzoic acid, 2,3-dihydroxybenzoic acid, 2,4-dihydroxybenzoic acid and 2,5-dihydroxybenzoic acid are not gallic acid decarboxylase substrates. A. adeninivorans G1212/YRC102-AYNI1-AGDC1, which expresses the AGDC1 gene under the control of the strong nitrate inducible AYNI1 promoter achieved a maximum gallic acid decarboxylase activity of 1064.4 U/l and 97.5 U/g of dry cell weight in yeast grown in minimal medium with nitrate as nitrogen source and glucose as carbon source. In the same medium, gallic acid decarboxylase activity was not detected for the control strain G1212/YRC102 with AGDC1 expression under the control of the endogenous promoter. Gene expression analysis showed that AGDC1 is induced by gallic acid and protocatechuic acid. In contrast to G1212/YRC102-AYNI1-AGDC1 and G1212/YRC102, A. adeninivorans G1234 [Δagdc1] is not able to grow on medium with gallic acid as carbon source but can grow in presence of protocatechuic acid. This confirms that Agdc1p plays an essential role in the tannic acid catabolism and could be useful in the production of catechol and cis,cis-muconic acid. However, the protocatechuic acid catabolism via Agdc1p to catechol seems to be not the only degradation pathway.


[A Portrait of Generalized Anxiety Symptoms in a Sample of High School Students: The Importance of Prevention, Screening and Intervention].
Anxiety disorders are among the most prevalent psychopathologies in children and adolescents around the world. They affect their personal, family, school and social functioning. Although symptoms of generalized anxiety disorder (GAD) are among the most commonly observed symptoms in adolescence, relatively little is known about the most important risk factors. Objectives The purpose of the present study was therefore 1) to document the prevalence of the central feature of GAD, excessive worrying, in high school students aged 12 to 17 years, by identifying the percentage of students reporting low, medium, and high levels of symptoms, 2) to identify key sociodemographic characteristics associated with high symptom level, and 3) to identify individual and family risk factors associated with high symptom level and estimate their relative contribution. Method A total of 8689 Quebec high school students (55.9% girls) participated in the study. The average age of these students was 14.34 years (SD=1.52). A k-means cluster analysis was first performed to create three categories based on the symptoms of generalized anxiety presented by the participants (low, medium, high). Multinomial logistic regression analyzes were then performed to identify the variables that best predict membership in these different categories. Results The results showed that 35.1% of the students reported a low level of symptoms, 40.2% reported an average level of symptoms, while 24.7% reported a high level of symptoms. Girls and adolescents in higher grades were more likely to report high levels of symptoms. Several factors were also identified as increasing the risk of being in the high generalized anxiety symptoms category compared to the average generalized anxiety symptoms and low generalized anxiety symptoms categories. These include fear of negative evaluation by others, perfectionism, depressed feelings, negative problem orientation and cognitive avoidance. Conclusion This study supports previous research showing that girls are at greater risk of developing generalized anxiety disorder. It also highlights the multidimensional aspect of the problem, by identifying the most important risk factors. Ultimately, this better understanding of the factors involved will allow us to better identify the targets to prioritize in the prevention of generalized anxiety disorders in adolescence.


A low cost kinect-based virtual rehabilitation system for inpatient rehabilitation of the upper limb in patients with subacute stroke: A randomized, double-blind, sham-controlled pilot trial.
We designed this study to prove the efficacy of the low-cost Kinect-based virtual rehabilitation (VR) system for upper limb recovery among patients with subacute stroke. A double-blind, randomized, sham-controlled trial was performed. A total of 23 subjects with subacute stroke (<3 months) were allocated to sham (n = 11) and real VR group (n = 12). Both groups participated in a daily 30-minute occupational therapy for upper limb recovery for 10 consecutive weekdays. Subjects received an additional daily 30-minute Kinect-based or sham VR. Assessment was performed before the VR, immediately and 1 month after the last session of VR. Fugl-Meyer Assessment (FMA) (primary outcome) and other secondary functional outcomes were measured. Accelerometers were used to measure hemiparetic upper limb movements during the therapy. FMA immediately after last VR session was not different between the sham (46.8 ± 16.0) and the real VR group (49.4 ± 14.2) (P = .937 in intention to treat analysis). Significant differences of total activity counts (TAC) were found in hemiparetic upper limb during the therapy between groups (F2,26 = 4.43; P = .22). Real VR group (107,926 ± 68,874) showed significantly more TACs compared with the sham VR group (46,686 ± 25,814) but there was no statistical significance between real VR and control (64,575 ± 27,533). Low-cost Kinect-based upper limb rehabilitation system was not more efficacious compared with sham VR. However, the compliance in VR was good and VR system induced more arm motion than control and similar activity compared with the conventional therapy, which suggests its utility as an adjuvant additional therapy during inpatient stroke rehabilitation.


'This town's a different town today':: Policing and regulating the night-time economy.
This article considers recent policing and regulatory responses to the night-time economy in England and Wales. Drawing upon the findings of a broader two-year qualitative investigation of local and national developments in alcohol policy, it identifies a dramatic acceleration of statutory activity, with 12 new or revised powers, and several more in prospect, introduced by the Labour Government within its first decade in office. Interview data and documentary sources are used to explore the degree to which the introduction of such powers, often accompanied by forceful rhetoric and high profile police action, has translated into a sustained expansion of control. Many of the new powers are spatially directed, as well as being focused upon the actions of distinct individuals or businesses, yet the willingness and capacity to apply powers to offending individuals in comparison to businesses is often variable and asymmetrical. The practice of negotiating order in the night-time economy is riddled with tensions and ambiguities that reflect the ad hoc nature and rapid escalation of the regulatory architecture. Night-time urban security governance is understood as the outcome of subtle organizational and interpersonal power-plays. Social orders, normative schemas and apportionments of blame thus arise as a byproduct of patterned (structural) relations.


Water Nanoconfined in a Hydrophobic Pore: Molecular Dynamics Simulations of Transmembrane Protein 175 and the Influence of Water Models.
Water molecules within biological ion channels are in a nanoconfined environment and therefore exhibit behaviors which differ from that of bulk water. Here, we investigate the phenomenon of hydrophobic gating, the process by which a nanopore may spontaneously dewet to form a "vapor lock" if the pore is sufficiently hydrophobic and/or narrow. This occurs without steric occlusion of the pore. Using molecular dynamics simulations with both rigid fixed-charge and polarizable (AMOEBA) force fields, we investigate this wetting/dewetting behavior in the transmembrane protein 175 ion channel. We examine how a range of rigid fixed-charge and polarizable water models affect wetting/dewetting in both the wild-type structure and in mutants chosen to cover a range of nanopore radii and pore-lining hydrophobicities. Crucially, we find that the rigid fixed-charge water models lead to similar wetting/dewetting behaviors, but that the polarizable water model resulted in an increased wettability of the hydrophobic gating region of the pore. This has significant implications for molecular simulations of nanoconfined water, as it implies that polarizability may need to be included if we are to gain detailed mechanistic insights into wetting/dewetting processes. These findings are of importance for the design of functionalized biomimetic nanopores (e.g., sensing or desalination) as well as for furthering our understanding of the mechanistic processes underlying biological ion channel function.


An orthogonal wavelet representation of multivalued images.
In this paper, a new orthogonal wavelet representation of multivalued images is presented. The idea for this representation is based on the concept of maximal gradient of multivalued images. This concept is generalized from gradients toward linear vector operators in the image plane with equal components along rows and columns. Using this generalization, the pyramidal dyadic wavelet transform algorithm using quadrature mirror filters is modified to be applied to multivalued images. This results in a representation of a single image, containing multiscale detail information from all component images involved. This representation leads to multiple applications ranging from multispectral image fusion to color and multivalued image enhancement, denoising and segmentation. In this paper, the representation is applied for fusion of images. More in particular, we introduce a scheme to merge high spatial resolution greylevel images with low spatial resolution multivalued images to improve spatial resolution of the latter while preserving spectral resolution. Two applications are studied: demosaicing of color images and merging of multispectral remote sensing images.


Activin treated urodele ectoderm: a model experimental system for cardiogenesis.
The tissue interactions which comprise the inductive phenomena associated with urodele heart morphogenesis are relatively well understood. In order to take full advantage of the experimental potential of this system formulation of an in vitro tissue culture system would be very helpful. Herein are described conditions for culturing Cynops pyrrhogaster early gastrula ectoderm tissue in the presence of the peptide growth factor activin. Two-week old explant cultures frequently displayed beating heart-like rudiments within. The beating frequency was measured and the extent to which cytodifferentiation mimicked normal heart differentiation assessed. Both measurements provided optimistic assessments which should encourage further exploitation of this model system.


Community Water Fluoridation in Focus: A Comprehensive Look at Fluoridation Levels across America.
Objective: This study reports on the number and percentage of community water systems (CWSs) meeting fluoride concentration standards set by the U.S. Department of Health and Human Services (DHHS). The study also explored changes in the population exposed to optimally fluoridated water in these systems between 2006 and 2020. Methods: This study analyzed U.S. Centers for Disease Control and Prevention data from 2006 to 2020, tabulating state-specific CWS fluoridation rates, ranking them, and calculating the percent change. Results: In 2020, 72.7% of the US population received CWS water, with 62.9% of those individuals served by a CWS system meeting DHHS fluoridation standards. This compares to 69.2% receiving CWS water in 2006 and 74.6% in 2012. The overall change in those receiving fluoridated water was 1.4%, from 61.5% in 2006 to 62.9% in 2020. State-specific percentages ranged from 8.5% in Hawaii to 100% in Washington DC in 2020 (median: 76.4%). Conclusions: Although endorsed by the American Dental Association, the percentage of individuals receiving fluoridated water did not increase substantially from 2006 to 2020, indicating that there has not been much progress toward meeting the Healthy People 2030 goal that 77.1% of Americans receive water with enough fluoride to prevent tooth decay.


Variation in diagnosis and treatment of chronic low back pain by traditional Chinese medicine acupuncturists.
To assess interrater reliability of Traditional Chinese Medicine (TCM) diagnosis and treatment of chronic low back pain. Under a Latin square design, six TCM acupuncturists evaluated the same six patients on the same day. Northwest Institute of Acupuncture and Oriental Medicine, Seattle, Washington. Assessment only. TCM diagnosis, acupoint prescriptions, auxiliary treatment recommendations. Twenty diagnoses and 65 acupoints were used at least once. The diagnosis of Qi/Blood Stagnation with Kidney Deficiency and the acupoint UB23 were used for every patient by most acupuncturists. However, consistency across acupuncturists regarding diagnostic details and other acupoints was poor. No diagnoses, and only one acupoint, were used preferentially for a subgroup of patients. Some diagnoses and treatment recommendations were dependent more on the practitioner than on the patient. Fine-grained diagnoses and most acupoints were unrelated to either patient or practitioner. TCM diagnoses and treatment recommendations for specific patients with chronic low back pain vary widely across practitioners. Acupuncture clinical trials using an individualized treatment arm may be difficult to replicate or evaluate because of low concordance among acupuncturists. Comparison of individualized treatment with a thoughtfully developed standardized approach is warranted to determine which, if either, is superior.


MRI of Superior Capsular Reconstruction.
Massive irreparable rotator cuff tears can be a challenging problem for arthroscopists in the perioperative setting because the typical treatment, reverse total shoulder arthroplasty, may not be the best option for all patients. Superior capsular reconstruction (SCR) is an advancing treatment option for patients with rotator cuff tears that are neither amenable to primary repair nor ideal for arthroplasty. Patient selection, which is strongly dependent on preoperative imaging findings, is an important step in obtaining favorable surgical outcomes. The tissue quality and tear type are particularly important when considering SCR for a patient. When unsuccessful SCR is suspected, postoperative MRI of the shoulder offers the surgeon and radiologist a means of evaluating the integrity and fixation of the graft. Fluid-sensitive MRI sequences are best for examining the final SCR construct, with high-signal-intensity fluid interruptions within the graft and the presence or worsening of shoulder arthropathy indicating graft failure. The indications for SCR are discussed, and the normal postoperative MRI findings after SCR are described in this review. In addition, the common types of SCR graft failure and associated imaging findings are described and illustrated. ©RSNA, 2020.


Maternal employment in a family context: effects on infant-mother and infant-father attachments.
The relation between resumption of full-time employment by mothers of infants under 6 months of age, and subsequent infant-mother and infant-father attachments, was examined in this study. Attachment classifications and ratings of reunion behavior with mother and with father in Ainsworth's Strange Situation at 12 months were obtained for 57 nonemployed-mother families and 40 employed-mother families. No relation emerged between maternal work status and the quality of infants' attachments to their mothers, indicating that early resumption of employment may not impede the development of secure infant-mother attachment. A significantly higher proportion of insecure attachments to fathers in employed-mother families was found for sons but not for daughters. Joint examination of the infants' attachments to both parents revealed a trend suggesting that in employed-mother families, boys were more likely to be insecurely attached to both parents than were girls in employed-mother families or infants of either sex in nonemployed-mother families. These patterns are discussed in light of differences in maternal and paternal sex-typing behavior and of evidence suggesting boys' vulnerability to psychosocial stress.


Trace elements assessment in Cerastoderma glaucum from port areas in the Tunisian Mediterranean coast: The influence of parasites on bioaccumulation.
In the present study, the seasonal concentration of seven trace elements was investigated in sediment and the cockle Cerastoderma glaucum tissues from two port areas in the North-East and South of Tunisia comparing cockles non-parasitized (NP) and parasitized (P) with digenean parasites. Elements concentration in sediments analyzed in both sites revealed that Zinc (Zn), Chromium (Cr), and Lead (Pb) were the most abundant ones, while Cadmium (Cd) and Mercury (Hg) were less abundant. The bioaccumulation of trace metals and Arsenic (As) in the tissues of cockles seems to be modulated by both the infection state and the parasite species. The relationship between bioaccumulation of metals and As, trematode species and abiotic parameters showed that the availability of certain metals for uptake by P cockles of both sites was influenced by the salinity and temperature of the water. Our results corroborate the possibility of using digenean infecting bivalves in biomonitoring aquatic ecosystems.


Acceptability of financial incentives and penalties for encouraging uptake of healthy behaviours: focus groups.
There is evidence that financial incentive interventions, which include both financial rewards and also penalties, are effective in encouraging healthy behaviours. However, concerns about the acceptability of such interventions remain. We report on focus groups with a cross-section of adults from North East England exploring their acceptance of financial incentive interventions for encouraging healthy behaviours amongst adults. Such information should help guide the design and development of acceptable, and effective, financial incentive interventions. Eight focus groups with a total of 74 adults were conducted between November 2013 and January 2014 in Newcastle upon Tyne, UK. Focus groups lasted approximately 60 minutes and explored factors that made financial incentives acceptable and unacceptable to participants, together with discussions on preferred formats for financial incentives. Verbatim transcripts were thematically coded and analysed in Nvivo 10. Participants largely distrusted health promoting financial incentives, with a concern that individuals may abuse such schemes. There was, however, evidence that health promoting financial incentives may be more acceptable if they are fair to all recipients and members of the public; if they are closely monitored and evaluated; if they are shown to be effective and cost-effective; and if clear health education is provided alongside health promoting financial incentives. There was also a preference for positive rewards rather than negative penalties, and for shopping vouchers rather than cash incentives. This qualitative empirical research has highlighted clear suggestions on how to design health promoting financial incentives to maximise acceptability to the general public. It will also be important to determine the acceptability of health promoting financial incentives in a range of stakeholders, and in particular, those who fund such schemes, and policy-makers who are likely to be involved with the design, implementation and evaluation of health promoting financial incentive schemes.


Risk to human health from a plethora of simian immunodeficiency viruses in primate bushmeat.
To assess human exposure to Simian immunodeficiency virus (SIV) in west central Africa, we looked for SIV infection in 788 monkeys that were hunted in the rainforests of Cameroon for bushmeat or kept as pets. Serologic reactivity suggesting SIV infection was found in 13 of 16 primate species, including 4 not previously known to harbor SIV. Overall, 131 sera (16.6%) reacted strongly and an additional 34 (4.3%) reacted weakly with HIV antigens. Molecular analysis identified five new phylogenetic SIV lineages. These data document for the first time that a substantial proportion of wild monkeys in Cameroon are SIV infected and that humans who hunt and handle bushmeat are exposed to a plethora of genetically highly divergent viruses.


Organic-Solvent-Tolerant Carboxylic Ester Hydrolases for Organic Synthesis.
Biocatalysis has emerged as an important tool in synthetic organic chemistry enabling the chemical industry to execute reactions with high regio- or enantioselectivity and under usually mild reaction conditions while avoiding toxic waste. Target substrates and products of reactions catalyzed by carboxylic ester hydrolases are often poorly water soluble and require organic solvents, whereas enzymes are evolved by nature to be active in cells, i.e., in aqueous rather than organic solvents. Therefore, biocatalysts that withstand organic solvents are urgently needed. Current strategies to identify such enzymes rely on laborious tests carried out by incubation in different organic solvents and determination of residual activity. Here, we describe a simple assay useful for screening large libraries of carboxylic ester hydrolases for resistance and activity in water-miscible organic solvents. We have screened a set of 26 enzymes, most of them identified in this study, with four different water-miscible organic solvents. The triglyceride tributyrin was used as a substrate, and fatty acids released by enzymatic hydrolysis were detected by a pH shift indicated by the indicator dye nitrazine yellow. With this strategy, we succeeded in identifying a novel highly organic-solvent-tolerant esterase from Pseudomonas aestusnigri In addition, the newly identified enzymes were tested with sterically demanding substrates, which are common in pharmaceutical intermediates, and two enzymes from Alcanivorax borkumensis were identified which outcompeted the gold standard ester hydrolase CalB from Candida antarcticaIMPORTANCE Major challenges hampering biotechnological applications of esterases include the requirement to accept nonnatural and chemically demanding substrates and the tolerance of the enzymes toward organic solvents which are often required to solubilize such substrates. We describe here a high-throughput screening strategy to identify novel organic-solvent-tolerant carboxylic ester hydrolases (CEs). Among these enzymes, CEs active against water-insoluble bulky substrates were identified. Our results thus contribute to fostering the identification and biotechnological application of CEs.


Cloning of TFC1, the Saccharomyces cerevisiae gene encoding the 95-kDa subunit of transcription factor TFIIIC.
The yeast gene encoding the 95-kDa subunit of the class III gene transcription factor TFIIIC was cloned. This gene, termed TFC1 (transcription factor C, gene 1), was isolated by screening a lambda gt11 yeast cDNA expression library using a polyclonal antiserum preparation which was previously shown to specifically recognize the 95-kDa subunit of yeast TFIIIC (Parsons, M. C., and Weil, P. A. (1990) J. Biol. Chem. 265, 5095-5103). TFC1 was found to be a single copy gene which contained a continuous open reading frame about 2 kilobases in length. TFC1 was shown to encode the 95-kDa subunit of TFIIIC by several criteria. Like the authentic yeast protein, the protein encoded by TFC1 had an apparent molecular weight of 95,000. In addition, the protein encoded by the TFC1 gene bound to the same antibody species as the yeast 95-kDa subunit of TFIIIC. Last, the sizes of the cleavage products of the Escherichia coli-expressed protein were indistinguishable from those of the cleavage products of the bona fide yeast 95-kDa protein.


[An ethological model of endogenous psychoses].
Our theory deals with the causality of the endogenous psychoses, considered as a shift of the systems of social behaviour that, in the natural history of our species, are supposed to have controlled the "Social homeostasis": territorial intolerance and dominance hierarchy. The subjective feeling of displacement beyond the boundaries of the group territory would trigger the paranoid syndrome, while the inner experience of an invasion of the own domain would lead to paranoia; hierarchical sudden ascent fits to the maniac syndromes, and the decay of dominance to depression. Intermingled emergence of territorial and ranking vectors allows us to obtain the syndrome profiles of mixed and atypical states with schizomorph and affective components. This causal model gives unity, as well as explanatory and comprehensive principles, to the clinical syndromes formed in the evolution of the endogenous psychoses.


Lopinavir impairs protein synthesis and induces eEF2 phosphorylation via the activation of AMP-activated protein kinase.
HIV anti-retroviral drugs decrease protein synthesis, although the underlying regulatory mechanisms of this process are not fully established. Therefore, we investigated the effects of the HIV protease inhibitor lopinavir (LPV) on protein metabolism. We also characterized the mechanisms that mediate the effects of this drug on elongation factor-2 (eEF2), a key component of the translational machinery. Treatment of C2C12 myocytes with LPV produced a dose-dependent inhibitory effect on protein synthesis. This effect was observed at 15 min and was maintained for at least 4 h. Mechanistically, LPV increased the phosphorylation of eEF2 and thereby decreased the activity of this protein. Increased phosphorylation of eEF2 was associated with increased activity of its upstream regulators AMP-activated protein kinase (AMPK) and eEF2 kinase (eEF2K). Both AMPK and eEF2K directly phosphorylated eEF2 in an in vitro kinase assay suggesting two distinct paths lead to eEF2 phosphorylation. To verify this connection, myocytes were treated with the AMPK inhibitor compound C. Compound C blocked eEF2K and eEF2 phosphorylation, demonstrating that LPV affects eEF2 activity via an AMPK-eEF2K dependent pathway. In contrast, incubation of myocytes with rottlerin suppressed eEF2K, but not eEF2 phosphorylation, suggesting that eEF2 can be regulated independent of eEF2K. Finally, LPV did not affect PP2A activity when either eEF2 or peptide was used as the substrate. Collectively, these results indicate that LPV decreases protein synthesis, at least in part, via inhibition of eEF2. This appears regulated by AMPK which can act directly on eEF2 or indirectly via the action of eEF2K.


Interstitial deletion 1p in a 30 year old woman.
High resolution chromosome analysis showed the karyotype 46,XX,del(1)(p22.1 p31.2) in a 30 year old woman with psychomotor retardation and various malformations. Determination of the enzyme phosphoglucomutase 1 (PGM1) showed that she was a heterozygote. Three other cases of interstitial deletion 1p have been reported previously, and one of these cases had several features in common with our case, suggesting a distinct syndrome.