Phytochrome control of specific mRNA levels in developing pea buds : the presence of both very low fluence and low fluence responses.
We have examined phytochrome regulated changes in transcript abundance for 11 different light regulated mRNAs in developing pea buds. Fluence-response curves were measured for changes in transcript abundance in response to red light pulses in both the low and very low fluence ranges. Most transcripts show only low fluence responses, with a threshold of approximately 10 micromoles per square meter. All of the low fluence responses are reversible by far red light. One transcript shows a very low fluence response, with a threshold of approximately 10(-4) micromoles per square meter. As expected, the very low fluence response is not far red reversible and in fact can be induced by far red light.Various fluences of red light were also used as pretreatments before transferring seedlings to continuous white light. One transcript responds to pretreatments in the very low fluence range, several respond to pretreatments in the low fluence range (including chlorophyll a/b binding protein RNA and ribulose-1,5-bisphosphate carboxylase RNA), and several show no response to the red light under these conditions. The threshold of these low fluence responses is approximately 10(2) micromoles per square meter, one order of magnitude greater than the threshold of the low fluence responses to red light alone.The transcripts may also be grouped by their responses to white light treatment alone. Three of the clones correspond to transcripts whose abundance decreases after a 24 hour white light treatment. The remainder of the mRNAs increase between 2- and 10-fold in response to the 24 hour white light.


Out-of-hospital cardiac arrest and percutaneous coronary intervention for ST-elevation myocardial infarction: long-term survival and neurological outcome.
Predictors of long-term outcome after ST-elevation myocardial infarction (STEMI) complicated by out-of-hospital cardiac arrest (OHCA) are incompletely understood, including the influence of successful coronary reperfusion. We analysed clinical and procedural data as well as 1-year outcome of 72 consecutive patients who underwent primary coronary intervention (PCI) after witnessed OHCA and STEMI and compared the results with 695 patients with STEMI and PCI, but without OHCA. Neurological recovery after OHCA was assessed using the Cerebral Performance Category (CPC) scale. PCI was successful in 83.3% after OHCA vs. 84.3% in the non-OHCA group (p=0.87). One-year mortality was 34.7% vs. 9.5% (p<0.001). 58.3% of the OHCA-patients showed complete neurological recovery (CPC 1) or moderate neurological disability (CPC 2). Another 6.9% showed severe cerebral disability (CPC 3) or permanent vegetative status (CPC 4). Delay from collapse until start of Advanced Cardiopulmonary Life Support (ACLS) was shorter for survivors with CPC status ≤2 (median 1 min, range 0-11 min) compared to non-survivors or survivors with CPC status >2 (median 8 min, range 0-13 min), p<0.0001. Age-adjusted multivariate analysis identified 'unsuccessful PCI', 'vasopressors on admission' and 'start of ACLS after >6 min' as independent predictors of negative long-term outcome (death or CPC >2). Mortality is high in patients with STEMI complicated by OHCA - even though PCI was performed with the same success rate as in patients without OHCA. The majority of survivors had favourable neurological outcomes at 1 year, especially if advanced life support had been started within ≤6 min and PCI was successful.


Screening effect of PEG on avidin binding to liposome surface receptors.
This study investigates the screening effect of poly(ethylene glycol)-phospholipids (PE-PEG) on the interaction of avidin with PEGylated liposomes containing surface-bound biotin ligands. The influence of grafting density and lipopolymer chain length is examined. A simple fluorescence assay involving a receptor-mediated fluorescence increase of BODIPY-labeled avidin upon binding to biotinylated lipids is employed to study the screening effect of submicellar concentrations of 1,2-dipalmitoyl-sn-glycero-3-phosphatidylethanolamine-N-[poly(ethylene glycol)-2000] (PE-PEG(2000)) and 1,2-dipalmitoyl-sn-glycero-3-phosphatidylethanolamine-N-[poly(ethylene glycol)-5000] (PE-PEG(5000)) incorporated into 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) liposomes. The results show that incorporation of lipopolymers into DPPC lipid bilayers reduces binding of avidin to the biotinylated liposomes, and it is found that the screening effect of PE-PEG(5000) is stronger than that for PE-PEG(2000). Thus, the results reveal that both the grafting density and the polymer length of the PE-PEG lipopolymers are of importance for the ability of water-soluble macromolecules to reach the surface of PEG liposomes. Furthermore, it is found that none of the lipopolymers completely prevents avidin from reaching the surface-bound biotin ligands.


Lifestyle change for coronary artery disease. What to tell patients.
Heart-healthy living is a wise choice for anyone, but it assumes even greater importance in persons who have or are at risk for coronary artery disease. Such patients need to know which measures have proved most effective in the prevention and treatment of the disease. This article reviews the data regarding risk factor reduction and offers recommendations for effective lifestyle modifications.


Partial restoration of defective chloride conductance in DeltaF508 CF mice by trimethylamine oxide.
This study was designed to test the in vivo efficacy of the chemical chaperone trimethylamine oxide (TMAO) in correcting the Cl- transport defect in a mouse model of cystic fibrosis (CF). Rectal potential difference (RPD) measurements were done in matched wild-type and DeltaF508 CF mice. Mice were treated by subcutaneous injections of TMAO. Wild-type mice demonstrated a forskolin-stimulated, Cl--dependent hyperpolarization of -6.4 +/- 0.8 mV (n = 11), which was significantly increased to -13.1 +/- 1.4 mV after treatment with TMAO. DeltaF508 CF mice showed no significant responses to forskolin. Treatment with TMAO recovered a forskolin-activated RPD in DeltaF508 CF mice (-1.1 +/- 0.2 mV; n = 17) but not in CFTR null mice. The effects of TMAO were dose dependent, resulting in a slope of -0.4 +/- 0.1 mV x g(-1) x kg(-1) in DeltaF508 CF mice. The forskolin-stimulated RPD in TMAO-treated DeltaF508 CF mice was partially blocked by glibenclamide and further stimulated by apigenin. The total response to forskolin plus apigenin was -2.5 +/- 0.45 mV (n = 6 mice), corresponding to 39% of the response evoked by forskolin only in wild-type mice.


Template- and pH-directed assembly of diruthenium diphosphonates with different topologies and oxidation states.
In the presence of organic templates, six diruthenium diphosphonates, namely, [H3N(CH2)3NH3]2[Ru2(hedp)2] (1), [H3N(CH2)4NH3]2[Ru2(hedp)2].4H2O (2), [H3N(CH2)5NH3]2[Ru2(hedp)2].4H2O (3), [H3N(CH2)3NH3][Ru2(hedp)(hedpH)].H2O (4), [H3N(CH2)4NH3][Ru2(hedpH(0.5))2].2H2O (5), and [H3N(CH2)5NH3]2[Ru2(hedp)2][Ru2(hedpH)2]] (6) [hedp = 1-hydroxyethylidenediphosphonate, CH3C(OH)(PO3)2] have been synthesized under hydrothermal conditions. Compounds 1-3 contain homovalent paddlewheel cores of Ru2(II,II)(hedp)2(4-) that are connected through edge-sharing of the [RuO5Ru] octahedra, resulting in infinite linear chains. Compounds 4-6 contain mixed-valent diruthenium(II,III) phosphonate paddlewheel cores of Ru2(II,III)(hedpH(n))2(3-2n)- that are connected by phosphonate oxygen atoms, forming distorted square-grid layers in 4 and 6 or a kagomé lattice in 5. Both the templates and the pH values are found to play important roles in directing the final products with particular topologies and oxidation states of the diruthenium unit. The magnetic studies show that weak antiferromagentic interactions are propagated between the homovalent diruthenium units in compounds 1-3. For compounds 4-6, weak ferromagnetic interactions are observed.


Physical health of autistic girls and women: a scoping review.
There is a growing recognition of sex and gender influences in autism. Increasingly, studies include comparisons between sexes or genders, but few have focused on clarifying the characteristics of autistic girls'/women's physical health. A scoping review was conducted to determine what is currently known about the physical health of autistic girls/women. We screened 1112 unique articles, with 40 studies meeting the inclusion criteria. We used a convergent iterative process to synthesize this content into broad thematic areas. Autistic girls/women experience more overall physical health challenges compared to non-autistic girls/women and to autistic boys/men. Emerging evidence suggests increased prevalence of epilepsy in autistic girls/women compared to non-autistic girls/women and to autistic boys/men. The literature also suggests increased endocrine and reproductive health conditions in autistic girls/women compared to non-autistic girls/women. Findings regarding gastrointestinal, metabolic, nutritional, and immune-related conditions are preliminary and inconsistent. The literature has substantial heterogeneity in how physical health conditions were assessed and reported. Further, our explicit focus on physical health may have constrained the ability to examine interactions between mental and physical health. The widely differing research aims and methodologies make it difficult to reach definitive conclusions. Nevertheless, in keeping with the goals of a scoping review, we were able to identify key themes to guide future research. The emerging literature suggests that autistic girls/women have heightened rates of physical health challenges compared to non-autistic girls/women and to autistic boys/men. Clinicians should seek to provide holistic care that includes a focus on physical health and develop a women's health lens when providing clinical care to autistic girls/women.


Isolated sphenoid sinus lesion: A diagnostic dilemma.
Isolated sphenoid sinus lesions are an uncommon entity and present with non-specific symptoms. In this case report, the patient presented with a history of headaches for a duration of one month without sinonasal symptoms. A computed tomography scan showed a soft tissue mass occupying the sphenoid sinus. An endoscopic biopsy revealed fungal infection. Endoscopic wide sphenoidotomy with excision of the sphenoid sinus lesion was then performed however, the microbiological examination post-surgery did not show any fungal elements. Instead, Citrobacter species was implicated to be the cause of infection.


Studies on Pea Ribosomal Proteins: Conformational and Biological Activity Changes of Ribosomal Subunits Derived by NH(4)Cl Dissociation.
Ribosomal subunits prepared by NH(4)Cl dissociation (0.5 m) of the monomeric ribosomes were much less active in in vitro protein synthesis than those prepared by KCl dissociation. The decrease in activity correlated with a detachment of some proteins (L(2) and L(9) as shown by gel electrophoresis) within the 60S ribosomal subunits. Subunits prepared with 0.3 m NH(4)Cl retained L(2) and L(9), but the activity remained low. Incubation of these 60S subunits in TKM buffer (50 mm tris [pH 7.5], 20 mm KCl, and 5 mm MgCl(2)) for 20 min at 37 C restored the activity almost to the level of those obtained by KCl dissociation. Treatment of the 0.3 m NH(4)Cl-derived 60S subunits with a protein reagent, Procion brilliant blue, prior to extraction of the ribosomal proteins resulted in the loss of L(2) and L(9), showing that these proteins were made accessible for dye binding. These observations suggest that a considerable degree of unfolding of the 60S subunit occurs at 0.3 m NH(4)Cl (this apparently leads to a preferential detachment of L(2) and L(9) at 0.5 m NH(4)Cl) and that the activity of the purified subunits depends not only on the presence of L(2) and L(9) but also on the organization of these proteins within the 60S subunits.


A decision aid for considering indomethacin prophylaxis vs. symptomatic treatment of PDA for extreme low birth weight infants.
Decision Aids (DA) are well established in various fields of medicine. It can improve the quality of decision-making and reduce decisional conflict. In neonatal care, and due to scientific equipoise, neonatologists caring for extreme low birth weight (ELBW) infants are in need to elicit parents' preferences with regard to the use of indomethacin therapy in ELBW infants. We aimed to develop a DA that elicits parents' preferences with regard to indomethacin therapy in ELBW infants. We developed a DA for the use of the indomethacin therapy in ELBW infants according to the Ottawa Decision Support Framework. The development process involved parents, neonatologists, DA developers and decision making experts. A pilot testing with healthy volunteers was conducted through an evaluation questionnaire, a knowledge scale, and a validated decisional conflict scale. The DA is a computer-based interactive tool. In the first part, the DA provides information about patent ductus arteriosus (PDA) as a disease, the different treatment options, and the benefits and downsides of using indomethacin therapy in preterm infants. In the second part, it coaches the parent in the decision making process through clarifying values and preferences. Volunteers rated 10 out of 13 items of the DA positively and showed significant improvement on both the knowledge scale (p = 0.008) and the decisional conflict scale (p = 0.008). We have developed a computer based DA to assess parental preferences with regard to indomethacin therapy in preterm infants. Future research will involve measurement of parental preferences to guide and augment the clinical decisions in current neonatal practice.


Prospective study of microchimerism in renal allograft recipients: association between HLA-DR matching, microchimerism and acute rejection.
The presence of donor-derived hematopoietic cells in blood and various tissues of the organ recipients, termed allogeneic microchimerism, has been considered to play an essential role in establishment of organ acceptance. In this study, we prospectively determined the presence of peripheral blood microchimerism (PBM) in 20 male-to-female renal allograft recipients up to 30 months post-transplantation. Recipients were categorized according to the pattern of microchimerism into microchimeric and nonmicrochimeric groups, and then state of human leukocyte antigens (HLA) Class II (DR/DQ) matching, episodes of acute rejection, age at transplantation, renal function, and history of blood transfusion were compared. DNA was extracted from donor, pre-transplant, and post-transplant (1 wk; 1, 3, 6, 12, 18, 24, and 30 months) peripheral blood samples. We analyzed PBM using nested polymerase chain reaction (PCR) amplification specific for the SRY region of the Y chromosome with a sensitivity up to 1:1 000 000. Microchimerism was detected in 13 (65%) of 20 recipients at various intervals. The highest frequency of microchimerism was at 1 wk (55%). Among microchimeric recipients, none were positive on all post-transplant analyses. Interestingly, nonmicrochimeric cases were negative throughout the study. The three recipients with an episode of acute rejection during the first week after transplantation were all in the nonmicrochimeric group with completely mismatched HLA-DR antigens. HLA-DR incompatibility was significantly lower (t-test, p<0.05) in microchimeric cases (1.0+/-0.58) than in nonmicrochimeric ones (1.9+/-0.38). But regarding HLA-DQ and other clinical parameters mentioned above, significant difference was not observed. We propose that there is an association between HLA-DR matching, microchimerism and acute graft rejection in our recipients. Our study demonstrates that, with routine immunosuppressive protocols, higher compatibility of HLA-DR antigens facilitates microchimerism induction. Then, development of new stronger immunosuppressive protocols (including conditioning) or augmentation of chimeric state (by donor-specific bone marrow infusion), especially in completely mismatched HLA-DR renal allograft recipients, may be useful for graft acceptance.


cAMP inducibility of transcriptional repressor ICER in developing and mature human T lymphocytes.
Stimulation of the cAMP-dependent signaling pathway exerts an inhibitory effect on the proliferation and effector functions of T cells. The ability of T cells to form high intracellular levels of cAMP is acquired during development in the human thymus and is retained by the majority of mature peripheral T lymphocytes. Here we show that elevated cAMP levels in T cells correlate with the expression of the potent transcriptional repressor ICER (inducible cAMP early repressor) previously described in the hypothalamic-pituitary-gonadal axis. Further, in transcriptional assays in vivo, ICER inhibits calcineurin-mediated expression of the interleukin 2 promoter as well as Tax-mediated transactivation of the human T-lymphotropic virus type I (HTLV-I) promoter. Thus, the induction of ICER in T cells may play an important role in the cAMP-induced quiescence and the persistent latency of HTLV-I.


Cadmium inhibits the functions of eukaryotic MutS complexes.
Exposure of yeast cells to low concentrations of cadmium results in elevated mutation rates due to loss of mismatch repair (MMR), and cadmium inhibits MMR activity in extracts of human cells. Here we show that cadmium inhibits both Msh2-Msh6- and Msh2-Msh3-dependent human MMR activity in vitro. This inhibition, which occurs at a step or steps preceding repair DNA synthesis, is observed for repair directed by either a 3' or a 5' nick. In an attempt to identify the protein target(s) of cadmium inhibition, we show that cadmium inhibition of MMR is not reversed by addition of zinc to the repair reaction, suggesting that the target is not a zinc metalloprotein. We then show that cadmium inhibits ATP hydrolysis by yeast Msh2-Msh6 but has no effect on ATPase hydrolysis by yeast Mlh1-Pms1. Steady state kinetic analysis with wild type Msh2-Msh6, and with heterodimers containing subunit-specific Glu to Ala replacements inferred to inactivate the ATPase activity of either Msh2 or Msh6, suggest that cadmium inhibits ATP hydrolysis by Msh6 but not Msh2. Cadmium also reduces DNA binding by Msh2-Msh6 and more so for mismatched than matched duplexes. These data indicate that eukaryotic Msh2-Msh3 and Msh2-Msh6 complexes are targets for inhibition of MMR by cadmium, a human lung carcinogen that is ubiquitous in the environment.


Fluid shear stress enhances T cell activation through Piezo1.
T cell activation is a mechanical process as much as it is a biochemical process. In this study, we used a cone-and-plate viscometer system to treat Jurkat and primary human T cells with fluid shear stress (FSS) to enhance the activation of the T cells through mechanical means. The FSS treatment of T cells in combination with soluble and bead-bound CD3/CD28 antibodies increased the activation of signaling proteins essential for T cell activation, such as zeta-chain-associated protein kinase-70 (ZAP70), nuclear factor of activated T cells (NFAT), nuclear factor kappa B (NF-κB), and AP-1 (activator protein 1). The FSS treatment also enhanced the expression of the cytokines tumor necrosis factor alpha (TNF-α), interleukin 2 (IL-2), and interferon gamma (IFN-γ), which are necessary for sustained T cell activation and function. The enhanced activation of T cells by FSS was calcium dependent. The calcium signaling was controlled by the mechanosensitive ion channel Piezo1, as GsMTx-4 and Piezo1 knockout reduced ZAP70 phosphorylation by FSS. These results demonstrate an intriguing new dynamic to T cell activation, as the circulatory system consists of different magnitudes of FSS and could have a proinflammatory role in T cell function. The results also identify a potential pathophysiological relationship between T cell activation and FSS, as hypertension is a disease characterized by abnormal blood flow and is correlated with multiple autoimmune diseases.


Explaining the effects of a decision intervention on mammography intentions: The roles of worry, fear and perceived susceptibility to breast cancer.
This study examines the effects of a mammography decision intervention on perceived susceptibility to breast cancer (PSBC) and emotion and investigates how these outcomes predict mammography intentions. Randomised between-subjects online experiment. Participants were stratified into two levels of risk. Within each stratum, conditions included a basic information condition and six decision intervention conditions that included personalised risk estimates and varied according to a 2 (amount of information: brief vs. extended) × 3 (format: expository vs. untailored exemplar vs. tailored exemplar) design. Participants included 2465 US women ages 35-49. PSBC as a percentage, PSBC as a frequency, worry, fear and mammography intentions. The intervention resulted in significant reductions in PSBC as a percentage for women in both strata and significant increases in worry and fear for women in the upper risk stratum. Of the possible mediators examined, only PSBC as a percentage was a consistent mediator of the effect of the intervention on mammography intentions. The results provide insight into the mechanism of action of the intervention by showing that PSBC mediated the effects of the intervention on mammography intentions.


A Small but Significantly Greater Incidence of Inflammatory Heart Disease Identified After Vaccination for Severe Acute Respiratory Syndrome Coronavirus 2.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines are being administered on an unprecedented scale. Assessing the risks of side effects is needed to aid clinicians in early detection and treatment. This study examined the risk of inflammatory heart disease, including pericarditis and myocarditis, after SARS-CoV-2 vaccination. Intermountain Healthcare patients with inflammatory heart disease from December 15, 2020 to June 15, 2021, and with or without preceding SARS-CoV-2 vaccinations, were studied. Relative rates of inflammatory heart disease were examined for vaccinated patients compared to unvaccinated patients. Of 67 patients identified with inflammatory heart disease, 21 (31.3%) had a SARS-Cov-2 vaccination within the previous 60 days. Overall, 914 611 Intermountain Healthcare patients received a SARS-CoV-2 vaccine, resulting in an inflammatory heart disease rate of 2.30 per 100 000 vaccinated patients. The relative risk of inflammatory heart disease for the vaccinated patients compared to the unvaccinated patients was 2.05 times higher rate within the 30-day window (P = .01) and had a trend toward increase in the 60-day window (relative rate = 1.63; P = .07). All vaccinated patients with inflammatory heart disease were treated successfully with 1 death related to a pre-existing condition. Although rare, the rate of inflammatory heart disease was greater in a SARS-CoV-2-vaccinated population than the unvaccinated population. This risk is eclipsed by the risk of contracting coronavirus disease 2019 and its associated, commonly severe outcomes. Nevertheless, clinicians and patients should be informed of this risk to facilitate earlier recognition and treatment.


Past, present, and future perspectives of pulmonary metastasectomy for patients with advanced colorectal cancer.
Over a half-century has passed since Thomford et al. proposed the selection criteria for pulmonary metastasectomy, and several prognostic factors have been identified. Although screening modalities and operations have changed dramatically, the important concepts of the selection criteria remain unchanged. Recent improvements in the survival outcomes of colorectal cancer patients undergoing pulmonary metastasectomy may be the result of strict adherence to the selection criteria for oligometastatic lung tumors, which can mimic local disease. Pulmonary metastasectomy has become an important option for selected patients with oligometastasis, based mainly on a large amount of retrospective data, but its effect on survival remains unclear. Curable pulmonary metastasis might be regarded as a "semi-local disease" under the spontaneous control of an acquired alteration in host immune status. The current practice of pulmonary metastasectomy for colorectal cancer focuses on selecting the most appropriate operation for selected patients. However, in the rapidly evolving era of immunotherapy, treatment-naïve patients for whom surgery is not suitable might be pre-conditioned by immunotherapy so that they may be considered for salvage surgery.


Hyperfocus: the forgotten frontier of attention.
'Hyperfocus' is a phenomenon that reflects one's complete absorption in a task, to a point where a person appears to completely ignore or 'tune out' everything else. Hyperfocus is most often mentioned in the context of autism, schizophrenia, and attention deficit hyperactivity disorder, but research into its effect on cognitive and neural functioning is limited. We propose that hyperfocus is a critically important aspect of cognition, particularly with regard to clinical populations, and that it warrants significant investigation. Hyperfocus, though ostensibly self-explanatory, is poorly defined within the literature. In many cases, hyperfocus goes undefined, relying on the assumption that the reader inherently knows what it entails. Thus, there is no single consensus to what constitutes hyperfocus. Moreover, some studies do not refer to hyperfocus by name, but describe processes that may be related. In this paper, we review how hyperfocus (as well as possibly related phenomena) has been defined and measured, the challenges associated with hyperfocus research, and assess how hyperfocus affects both neurotypical and clinical populations. Using this foundation, we provide constructive criticism about previously used methods and analyses. We also propose an operational definition of hyperfocus for researchers to use moving forward.


Methodological aspects of outcomes research.
A critical evaluation of existing scientific evidence of treatment efficacy can be an important part of communicating risk and benefits of treatment options to patients during the course of clinical practice. A checklist of key methodological issues to examine when reading a research study is presented and discussed. Steps in reading a paper include: identifying the research question; identifying the manner in which subjects get enrolled in the study; identifying the treatments and outcomes used; identifying the study design and the comparisons being made; evaluating the study methods for the possibility of bias and uncontrolled confounding; assessing whether the statistical analysis used is appropriate for the study design; assessing whether the study has sufficient statistical power to demonstrate hypotheses being tested. Finally, procedures for grading and evaluating evidence, as used by systematic review groups and international best evidence synthesis consensus groups is briefly described.


Optimization by infusion of multiple reaction monitoring transitions for sensitive quantification of peptides by liquid chromatography/mass spectrometry.
In peptide quantification by liquid chromatography/mass spectrometry (LC/MS), the optimization of multiple reaction monitoring (MRM) parameters is essential for sensitive detection. We have compared different approaches to build MRM assays, based either on flow injection analysis (FIA) of isotopically labelled peptides, or on the knowledge and the prediction of the best settings for MRM transitions and collision energies (CE). In this context, we introduce MRMOptimizer, an open-source software tool that processes spectra and assists the user in selecting transitions in the FIA workflow. MS/MS spectral libraries with CE voltages from 10 to 70 V are automatically acquired in FIA mode for isotopically labelled peptides. Then MRMOptimizer determines the optimal MRM settings for each peptide. To assess the quantitative performance of our approach, 155 peptides, representing 84 proteins, were analysed by LC/MRM-MS and the peak areas were compared between: (A) the MRMOptimizer-based workflow, (B1) the SRMAtlas transitions set used 'as-is'; (B2) the same SRMAtlas set with CE parameters optimized by Skyline. 51% of the three most intense transitions per peptide were shown to be common to both A and B1/B2 methods, and displayed similar sensitivity and peak area distributions. The peak areas obtained with MRMOptimizer for transitions sharing either the precursor ion charge state or the fragment ions with the SRMAtlas set at unique transitions were increased 1.8- to 2.3-fold. The gain in sensitivity using MRMOptimizer for transitions with different precursor ion charge state and fragment ions (8% of the total), reaches a ~ 11-fold increase. Isotopically labelled peptides can be used to optimize MRM transitions more efficiently in FIA than by searching databases. The MRMOptimizer software is MS independent and enables the post-acquisition selection of MRM parameters. Coefficients of variation for optimal CE values are lower than those obtained with the SRMAtlas approach (B2) and one additional peptide was detected. Copyright © 2017 John Wiley & Sons, Ltd.


Evaluation of the β-Glucuronidase Substrate 5-Bromo-4-Chloro-3-Indolyl-β-D-Glucuronide (X-GLUC) in a 24-Hour Direct Plating Method for Escherichia coli.
A 24-h direct plating method for Escherichia coli using Peptone-Tergitol agar was used to compare the effectiveness of the chromogenic substrate 5-bromo-4-chloro-3-indolyl-β-D-glucuronide (X-GLUC) with the fluorogenic substrate 4-methylumbelliferyl-β-D-glucuronide (MUG) for β-glucuronidase activity. Values obtained for enumeration of two strains of E. coli recovered from artificially inoculated raw minced chicken (i.e., plating efficiencies on the inoculum, cells per g, and recovery percentages elated to those on Plate Count Agar) indicate that X-GLUC at 50 μg/ml was as effective as MUG in an agar medium. Unlike MUG, X-GLUC does not require ultraviolet light illumination, and the color reaction produced remains localized in the positive colonies.


The recent Italian Consolidation Act on privacy: new measures for data protection.
In the light of the new Consolidation Act on privacy, in effect since January 2004, the author has made some observations in relation to the protection of data concerning health. Special focus has been given to an overall view of the new regulations. "Sensitive data" are seen as differing from health data, while always respecting the rights, the fundamental freedom and the dignity of the person involved. A central role continues to be played by the Controller, who has regulating and control powers.


[Determination of four fractions of Lycium barbarum polysaccharides in different varieties].
The contents of four fractions of Lycium barbarum polysaccharides (LBP) in different varieties were analysed. This method can be used for quality evaluation of Fractus Lycii.


Essential Role of Visfatin in Lipopolysaccharide and Colon Ascendens Stent Peritonitis-Induced Acute Lung Injury.
Acute lung injury (ALI) is a life-threatening syndrome characterized by acute and severe hypoxemic respiratory failure. Visfatin, which is known as an obesity-related cytokine with pro-inflammatory activities, plays a role in regulation of inflammatory cytokines. The mechanisms of ALI remain unclear in critically ill patients. Survival in ALI patients appear to be influenced by the stress generated by mechanical ventilation and by ALI-associated factors that initiate the inflammatory response. The objective for this study was to understand the mechanisms of how visfatin regulates inflammatory cytokines and promotes ALI. The expression of visfatin was evaluated in ALI patients and mouse sepsis models. Moreover, the underlying mechanisms were investigated using human bronchial epithelial cell lines, BEAS-2B and NL-20. An increase of serum visfatin was discovered in ALI patients compared to normal controls. Results from hematoxylin and eosin (H&E) and immunohistochemistry staining also showed that visfatin protein was upregulated in mouse sepsis models. Moreover, lipopolysaccharide (LPS) induced visfatin expression, activated the STAT3/NFκB pathway, and increased the expression of pro-inflammatory cytokines, including IL1-β, IL-6, and TNF-α in human bronchial epithelial cell lines NL-20 and BEAS-2B. Co-treatment of visfatin inhibitor FK866 reversed the activation of the STAT3/NFκB pathway and the increase of pro-inflammatory cytokines induced by LPS. Our study provides new evidence for the involvement of visfatin and down-stream events in acute lung injury. Further studies are required to confirm whether the anti-visfatin approaches can improve ALI patient survival by alleviating the pro-inflammatory process.


Effect of probiotics and synbiotics on blood glucose: a systematic review and meta-analysis of controlled trials.
High fasting blood glucose (FBG) can lead to chronic diseases such as diabetes mellitus, cardiovascular and kidney diseases. Consuming probiotics or synbiotics may improve FBG. A systematic review and meta-analysis of controlled trials was conducted to clarify the effect of probiotic and synbiotic consumption on FBG levels. PubMed, Scopus, Cochrane Library, and Cumulative Index to Nursing and Allied Health Literature databases were searched for relevant studies based on eligibility criteria. Randomized or non-randomized controlled trials which investigated the efficacy of probiotics or synbiotics on the FBG of adults were included. Studies were excluded if they were review articles and study protocols, or if the supplement dosage was not clearly mentioned. A total of fourteen studies (eighteen trials) were included in the analysis. Random-effects meta-analyses were conducted for the mean difference in FBG. Overall reduction in FBG observed from consumption of probiotics and synbiotics was borderline statistically significant (-0.18 mmol/L 95 % CI -0.37, 0.00; p = 0.05). Neither probiotic nor synbiotic subgroup analysis revealed a significant reduction in FBG. The result of subgroup analysis for baseline FBG level ≥7 mmol/L showed a reduction in FBG of 0.68 mmol/L (-1.07, -0.29; ρ < 0.01), while trials with multiple species of probiotics showed a more pronounced reduction of 0.31 mmol/L (-0.58, -0.03; ρ = 0.03) compared to single species trials. This meta-analysis suggests that probiotic and synbiotic supplementation may be beneficial in lowering FBG in adults with high baseline FBG (≥7 mmol/L) and that multispecies probiotics may have more impact on FBG than single species.


Blood flow in the human iris measured by laser Doppler flowmetry.
A new instrument based on laser Doppler flowmetry (LDF) has been developed to determine noninvasively the relative flux of red blood cells (RBCs) through the microcirculatory network of the iris of the human eye. The probing laser, photodetector, and target fixation devices required by this method were adapted to a slit lamp. Electronic processing and computer analysis of the Doppler signal allow determination of relative velocity, number, and flux of RBCs in the iris, as well as the pulsatility of these flow parameters during the heart cycle. Based on measurements in one eye of eight normal volunteers, the sensitivity of the technique, i.e., the minimum change detectable at the P < 0.05 level, was 4% for the flux. The decrease in blood flow in response to decreases of the ocular perfusion pressure demonstrates the capability of the technique to detect flow changes and its suitability for investigating the physiology and the pharmacology of iris circulation.


Iodine-125 induced cholangiocarcinoma cell death is enhanced by inhibition of endoplasmic reticulum stress-mediated protective autophagy.
Cholangiocarcinoma (CCA) is the second most common primary liver malignancy, however, it is difficult to diagnose and treat, and only a few patients with CCA are suitable for surgery. Iodine-125 (I-125) is an effective treatment for cancer, but the molecular mechanisms underlying the effects of I-125 differ among different cancers. This study aimed to explore the effects of I-125 on CCA cell activity and determine the possible mechanisms of action of I-125 in this type of cancer. CCA cell proliferation, cycling, apoptosis, autophagy, and endoplasmic reticulum (ER) stress were determined after irradiation of CCA cells with I-125 seeds. The effects of I-125 on autophagy and ER stress in three CCA cell lines were evaluated using western blotting, while the effects of I-125 on apoptosis and autophagy in QBC939 cells treated with si-Beclin1 or si-PERK, respectively, were assessed using flow cytometry. I-125 suppressed cell viability and induced cell cycle G2/M-phase arrest in three CCA cell lines (QBC939, TFK-1, HuCCT1). I-125 induced apoptosis, autophagy, and ER stress by altering the expression levels of some related proteins in each of the three CCA cell lines. Furthermore, autophagy inhibition (treatment with si-Beclin1) increased expression of apoptosis-related proteins (cleaved-PARP and cleaved-caspase-3, Bax/Bcl2) in QBC939 cells irradiated with I-125 seeds, while ER stress inhibition (with si-PERK) suppressed the expression of autophagy-related proteins (LC3-I, LC3-II, p62). Therefore, I-125 induces ER stress, thereby activating protective autophagy in CCA cells through the PERK signaling pathway. Combined inhibition of ER stress and autophagy signaling may increase the killing effect of I-125 on cancer cells and serve as a new auxiliary method in I-125 radiotherapy.


A new canine model to evaluate the biological response of intramedullary bone to implant materials and surfaces.
A new canine model utilizing an implantable chamber with multiple bone ingrowth channels has been used to study the response of intramedullary bone to various implant materials and surfaces. The first group of dogs received implants containing channels lined by smooth-surfaced coupons of titanium, titanium alloy, sputter-hydroxyapatite-coated (HA-coated) titanium alloy, and polyethylene. A pattern of early initial bone ingrowth by 2 weeks, becoming maximal at 6 to 12 weeks with remodeling to a more mature lamellar bone, and later resorption by 24 weeks was seen for all test groups, with fibrous tissue interfaces covering the smooth test coupons at all time points. Significantly increased bone ingrowth in the sputter-HA coated group was found only at 6 weeks. The second group of dogs received implants with channels lined by surface-roughened coupons of either titanium or plasma-HA-coated titanium, half of which were also packed with a crystalline-HA grouting at the time of surgery. At both 6 and 12 weeks, bone ingrowth was greatly enhanced by the presence of the plasma-HA coating or the crystalline-HA grouting as compared to the uncoated titanium channels. Histologically, bone was seen to bond directly to the plasma-HA coating and the crystalline-HA grouting. A thin fibrous tissue layer was noted between bone and the titanium in most areas, but evidence of direct bone contact to the metal surface was seen. Mechanical testing in tension of intact coupon-bone-coupon units revealed significant strength of the bone-plasma-HA bond, with failure initiating at the metal-HA interface with forces of 15.3 N at 6 weeks, increasing to 44.8 N at 12 weeks. Plasma-HA-lined channels with crystalline-HA packing required similar forces for failure. No significant adhesion strength was noted for the titanium channels at 6 weeks, and only the crystalline-HA-filled channels displayed measurable strength of the bone-titanium interface at 12 weeks, with a force of 9 N needed for failure.


Assessing the Link Between Perception and Production in Cantonese Tone Acquisition.
Purpose Previous studies showed early production precedes late perception in Cantonese tone acquisition, contrary to the general principle that perception precedes production in child language. How tone production and perception are linked in 1st language acquisition remains largely unknown. Our study revisited the acquisition of tone in Cantonese-speaking children, exploring the possible link between production and perception in 1st language acquisition. Method One hundred eleven Cantonese-speaking children aged between 2;0 and 6;0 (years;months) and 10 adolescent reference speakers participated in tone production and perception experiments. Production materials with 30 monosyllabic words were transcribed in filtered and unfiltered conditions by 2 native judges. Perception accuracy was based on a 2-alternative forced-choice task with pictures covering all possible tone pair contrasts. Results Children's accuracy of production and perception of all the 6 Cantonese tones was still not adultlike by age 6;0. Both production and perception accuracies matured with age. A weak positive link was found between the 2 accuracies. Mother's native language contributed to children's production accuracy. Conclusions Our findings show that production and perception abilities are associated in tone acquisition. Further study is needed to explore factors affecting production accuracy in children. Supplemental Material https://doi.org/10.23641/asha.7960826.


CD62L as target receptor for specific gene delivery into less differentiated human T lymphocytes.
Chimeric antigen receptor (CAR)-expressing T cells are a complex and heterogeneous gene therapy product with variable phenotype compositions. A higher proportion of less differentiated CAR T cells is usually associated with improved antitumoral function and persistence. We describe in this study a novel receptor-targeted lentiviral vector (LV) named 62L-LV that preferentially transduces less differentiated T cells marked by the L-selectin receptor CD62L, with transduction rates of up to 70% of CD4+ and 50% of CD8+ primary T cells. Remarkably, higher amounts of less differentiated T cells are transduced and preserved upon long-term cultivation using 62L-LV compared to VSV-LV. Interestingly, shed CD62L neither altered the binding of 62L-LV particles to T cells nor impacted their transduction. The incubation of 2 days of activated T lymphocytes with 62L-LV or VSV-LV for only 24 hours was sufficient to generate CAR T cells that controlled tumor growth in a leukemia tumor mouse model. The data proved that potent CAR T cells can be generated by short-term ex vivo exposure of primary cells to LVs. As a first vector type that preferentially transduces less differentiated T lymphocytes, 62L-LV has the potential to circumvent cumbersome selections of T cell subtypes and offers substantial shortening of the CAR T cell manufacturing process.


Cloning and sequence analysis of Wild Argali short palate, lung and nasal epithelium clone 1 cDNA.
Experiments were conducted to clone the sequence of Wild Argali short palate, lung and nasal epithelium clone 1 (SPLUNC1) cDNA, and to lay the foundation for further study the biological function of Wild Argali SPLUNC1. The complete sequence of Wild Argali SPLUNC1 cDNA was generated by rapid amplification of cDNA ends. The entire coding sequence was inserted into the pPIC9K vector and expressed in Pichia pastoris (P. pastoris) GS115. The recombinant SPLUNC1 protein was detected by Western blot and purified by Ni2+ chelate affinity chromatography. The test of effect of the protein on Mycoplasma ovipneumoniae (MO) was performed with real-time polymerase chain reaction. The Wild Argali SPLUNC1 cDNA was 1,076 bp with an open reading frame of 768 bp, which encoded a 26.49 kDa protein composed of 255 amino acids. Its amino acid sequence shared 98.4%, 96.9%, 94.5%, 90.2%, 80.8%, 78.4%, 78.3%, 72.5%, 72.3%, 68.8% identity with those of SPLUNC1 cDNA from Ovis aries (accession no. NP_001288334.1), Capra hircus (accession no. XP_005688516.1), Pantholops hodgsonii (accession no. XP_005979709.1), Bos taurus (accession no. NP_776851.1), Felis catus (accession no. XP_006929910.1), Homo sapiens (accession no. NP_001230122.1), Sus scrofa (accession no. NP_001005727.1), Chinchilla lanigera (accession no. NP_001269294.1), Mus musculus (accession no. NP_035256.2), and Rattus norvegicus (accession no. NP_742028.1), respectively. The recombinant protein corresponded to the expected molecular mass of 25.47 kDa as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and it was detected in the supernatant of P. pastoris, and it could be purified. The results from the test of inhibition effect of argali recombinant SPLUNC1 protein on MO showed that the product could inhibit MO very well (p<0.01). The amino acid sequence of Wild Argali SPLUNC1 was different from other organisms. The recombinant SPLUNC1 protein has good biological activity.


Expression of high affinity receptors for erythropoietin on human bone marrow cells and on the human erythroleukemic cell line, HEL.
The principal growth factor involved in the regulation of erythropoiesis, erythropoietin (Epo), is currently under clinical trial for the treatment of anemia. Despite the advanced state of these trials, little is known about the nature and distribution of the receptor for Epo on human hemopoietic cells or about the cellular mechanisms of signal transduction. In the present study 125I-labeled recombinant human Epo has been used to demonstrate expression of saturable, high affinity binding sites for Epo on density-fractionated human bone marrow cells and on the human erythroleukemic cell line, HEL. Binding was reversible and proportional to cell number, and HEL cells were shown to express on average 34 receptors per cell (range 30-35) with an affinity of 293 pM (range 275-300 pM) at 37 degrees C in the presence of sodium azide to block receptor internalization. Autoradiographic analysis of Epo binding to human bone marrow cells showed that specific binding, measured as the difference in grain counts between total binding and binding in the presence of excess unlabeled Epo, was greatest to pronormoblasts and declined during erythroid cell maturation to undetectable levels on nucleated red cells. Autoradiography also revealed significant Epo binding to marrow megakaryocytes, which comprise less than 1% of nonerythroid cells. In contrast to erythroid cells, Epo binding to megakaryocytes increased with cell maturation, with stage IV megakaryocytes exhibiting the highest specific binding. Grain density per surface area however, remained constant during megakaryocyte maturation and was approximately 25% that on pronormoblasts.


An eosinophil chemotactic lymphokine in the supernatant of lymphokine-activated killer (LAK) cell culture: relationship to interleukin-5.
The administration of interleukin-2 (IL-2) systemically or locally to cancer patients, either alone or in combination with lymphokine-activated killer (LAK) cells, frequently results in eosinophilia. To investigate the mechanism of such eosinophil accumulation, we measured in vitro eosinophil chemotactic activity (ECA) in the supernatant of LAK cell cultures by a modified Boyden's chamber method. Consequently, a potent and specific ECA was found in the cell-free supernatant of LAK cell cultures. On the other hand, no ECA was found in the supernatant of normal lymphocyte cultures, nor in IL-2 itself. This activity plateaued when 3 x 10(6) lymphocytes were incubated with 1000 U/ml IL-2 for more than 3 days. Sephadex G-100 gel chromatography indicated that the molecular weight of this eosinophil chemotactic factor (ECF) was approximately 45,000 Da. The ECF was stable when heated to 56 degrees C for 30 min but was inactivated by trypsin, indicating that the ECF is a lymphokine which has very similar characteristics to those of IL-5. Using ELISA for hIL-5, the IL-5 level in the supernatant of LAK cell cultures increased dose dependently with increasing IL-2 concentrations. These results suggest that IL-5 may be responsible for the local recruitment of eosinophils in IL-2/LAK therapy.


Enzyme activities associated with carcinogen metabolism in liver and nonhepatic tissues of rats maintained on high fat and food-restricted diets.
The influence of high fat or food-restricted diets on key enzymes associated with polycyclic aromatic hydrocarbon metabolism was assessed in liver, lung, kidney and stomach of rats. Animals had access ad libitum to the AIN-76A purified diet (control) or were given 65% of the intake of controls for 3 wk. The high fat diet was isoenergetic to the control diet by substituting corn oil for equal energy from carbohydrate and addition of cellulose to obtain equal energy density. Activities of arylhydrocarbon hydroxylase and 7-ethoxycoumarin O-deethylase were significantly increased in lungs of food-restricted rats and decreased by the high fat diet but were not altered in liver. Both diets increased arylhydrocarbon hydroxylase approximately twofold in kidney. Glucose 6-phosphate and 6-phosphogluconate dehydrogenase were lowered in lung, kidney and liver by the high fat diet. Hepatic glutathione S-transferase was increased by high fat feeding. Food restriction decreased activities of arylsulfatase and beta-glucuronidase about 40% in lung. Hepatic arylsulfatase was also decreased about 40% by this treatment. Changes in hydrolase activities in livers and lungs of animals maintained on restricted diets raises in the interesting possibility that net production of glucuronide and sulfate conjugates of carcinogens by the liver and their hydrolysis in lung is altered by food restriction.


A Deep Learning Framework for Spatiotemporal Ultrasound Localization Microscopy.
Ultrasound Localization Microscopy (ULM) can resolve the microvascular bed down to a few micrometers. To achieve such performance, microbubble contrast agents must perfuse the entire microvascular network. Microbubbles are then located individually and tracked over time to sample individual vessels, typically over hundreds of thousands of images. To overcome the fundamental limit of diffraction and achieve a dense reconstruction of the network, low microbubble concentrations must be used, which leads to acquisitions lasting several minutes. Conventional processing pipelines are currently unable to deal with interference from multiple nearby microbubbles, further reducing achievable concentrations. This work overcomes this problem by proposing a Deep Learning approach to recover dense vascular networks from ultrasound acquisitions with high microbubble concentrations. A realistic mouse brain microvascular network, segmented from 2-photon microscopy, was used to train a three-dimensional convolutional neural network (CNN) based on a V-net architecture. Ultrasound data sets from multiple microbubbles flowing through the microvascular network were simulated and used as ground truth to train the 3D CNN to track microbubbles. The 3D-CNN approach was validated in silico using a subset of the data and in vivo in a rat brain. In silico, the CNN reconstructed vascular networks with higher precision (81%) than a conventional ULM framework (70%). In vivo, the CNN could resolve micro vessels as small as 10 μ m with an improvement in resolution when compared against a conventional approach.


Alternative nitrogenase.
Recently, it has been demonstrated that both A. vinelandii and A. chroococcum have the ability to synthesize several different nitrogen-fixing enzymes. Both species can produce a Mo- or V-containing nitrogenase while A. vinelandii can also generate an all-Fe form of the enzyme. Regardless of the source or form of the enzyme, all nitrogenases are composed of two separable proteins, called components 1 and 2, where component 2 is a highly conserved dimer containing a single [4Fe-4S] cluster. The major differences among the various forms of the enzyme are in component 1, the protein where substrate reduction occurs. This protein can exist in forms of four, five, or six subunits and can contain Mo, V or only Fe at the putative active site. Finally, there are also variations among the different enzyme systems regarding the paramagnetism of component 1 as well as the general substrate reduction patterns. While it is not yet known why these bacteria possess the ability to generate multiple forms of nitrogenase or which form of the enzyme has the greatest physiological importance, the existence of these various nitrogenases raises several other important questions. Specifically, how do Mo, V and Fe regulate the synthesis of each enzyme system, which genes are unique and which are common for these systems, and what roles, if any, do Mo, V and Fe play in catalysis? The fact that we can now use three different enzyme systems to investigate nitrogen fixation may greatly increase our ability to answer these questions and eventually understand the mechanism of this very important enzymatic reaction.


Two-Pore-Domain Potassium (K2P-) Channels: Cardiac Expression Patterns and Disease-Specific Remodelling Processes.
Two-pore-domain potassium (K2P-) channels conduct outward K+ currents that maintain the resting membrane potential and modulate action potential repolarization. Members of the K2P channel family are widely expressed among different human cell types and organs where they were shown to regulate important physiological processes. Their functional activity is controlled by a broad variety of different stimuli, like pH level, temperature, and mechanical stress but also by the presence of lipids or pharmacological agents. In patients suffering from cardiovascular diseases, alterations in K2P-channel expression and function have been observed, suggesting functional significance and a potential therapeutic role of these ion channels. For example, upregulation of atrial specific K2P3.1 (TASK-1) currents in atrial fibrillation (AF) patients was shown to contribute to atrial action potential duration shortening, a key feature of AF-associated atrial electrical remodelling. Therefore, targeting K2P3.1 (TASK-1) channels might constitute an intriguing strategy for AF treatment. Further, mechanoactive K2P2.1 (TREK-1) currents have been implicated in the development of cardiac hypertrophy, cardiac fibrosis and heart failure. Cardiovascular expression of other K2P channels has been described, functional evidence in cardiac tissue however remains sparse. In the present review, expression, function, and regulation of cardiovascular K2P channels are summarized and compared among different species. Remodelling patterns, observed in disease models are discussed and compared to findings from clinical patients to assess the therapeutic potential of K2P channels.


A comprehensive comparative review of adenosine diphosphate receptor antagonists.
Thrombosis risk necessitates dual antiplatelet therapy with aspirin and an adenosine diphosphate (ADP) receptor antagonist, in patients who have acute coronary syndrome. Current guidelines emphasize the critical role of dual antiplatelet therapy in both medical management and invasive strategy, especially in patients undergoing percutaneous coronary intervention. With the availability of multiple ADP-receptor antagonists, it is crucial to select the most appropriate agent for each patient. The pertinent trials were identified through a MEDLINE search, in addition to a manual search from the articles retrieved. This review examines the differences between clopidogrel, prasugrel and ticagrelor in terms of their pharmacological/pharmacokinetic properties, clinical efficacy, drug interactions and safety parameters. Prasugrel and ticagrelor exhibit greater platelet inhibition and superior efficacy compared with clopidogrel, at the expense of higher bleeding risk. Prasugrel and ticagrelor should be preferred over clopidogrel in patients who are at a high risk of thrombotic events with low risk of bleeding. Additionally, these two agents may offer advantage over clopidogrel in those patients who might have risk for drug resistance due to CYP2C19 polymorphism. In selecting the ideal agent for patients, clinicians should tailor the antiplatelet regimen by considering individual risk factors and medication characteristics.


Effect of light source and specimen thickness on the surface hardness of resin composite.
To compare the Knoop hardness of different thicknesses of resin composite irradiated using either a plasma arc curing (PAC) light, or a quartz tungsten halogen (QTH) light. A 2.5-mm diameter hole was drilled through the center of 2, 3, 4, and 5-mm thick discs of enamel and dentin. This hole was filled with either Filtek P60 or Prodigy Condensable composite and irradiated in bulk using: (a) a PAC light with an 8-mm, 470 nm light guide for 3 seconds, (b) a QTH light with an 8-mm standard light guide for 40 seconds (QTH+S), or (c) a QTH light with a 13/8-mm Turbo light guide for 40 seconds (QTH+T). Immediately after irradiating the composite, the surface hardness was measured at the bottom and then the top. Hardness values were remeasured after 24-hour and 7-day storage in water at 37 degrees C. The hardness values were compared using the Analysis of Variance for Repeated Measures and Least Squares Means test for multiple comparisons (LSM test) at P< 0.05. The power density delivered by each light was: PAC over 1999 mW/cm2, QTH+S 588 mW/cm2, and QTH+T 844 mW/cm2. Both the thickness of the composite and the light source had a significant effect on the hardness. Overall, using the QTH+T produced the highest hardness values and the PAC light the lowest values. For all 2-mm thick specimens, irrespective of the light source, the hardness values at the bottom were not significantly different from their top hardness values measured at the same time. For the 3-mm thick specimens using the QTH+T, the hardness values at the bottom of both composites were not significantly different from their top hardness values at all time intervals. For the 3-mm thick specimens using the QTH+S, the bottom hardness values were not significantly different from those at the top for P60 only, and only at 24 hours and 7 days, but not when measured immediately. For all the 4 and 5-mm thick specimens, the hardness values at the bottom were all significantly different from those at the top measured at the same time interval.


Strength of the respiratory and lower limb muscles and functional capacity in chronic stroke survivors with different physical activity levels.
The assessment of strength and its relationships with functional capacity could contribute to more specific and effective disability management of stroke survivors. To compare and investigate associations between measures of strength and functional capacity of 98 chronic stroke survivors, stratified into three groups, according to their physical activity levels. The physical activity levels were classified as impaired, moderately active, and active, based on their Human Activity Profile (HAP) scores. Strength was assessed by the maximal inspiratory (MIP) and expiratory (MEP) pressures and by the residual deficits (RDs) of work of the lower limb and trunk muscles, whereas functional capacity was evaluated by the distance covered during the six-minute walking test (6MWT). One-way analyses of variance revealed significant differences between the groups, except between the active and moderately active groups regarding the RDS of the hip and knee flexors/extensors and ankle dorsiflexors (2.91<F<8.62; 0.001<p<0.01). Differences between the groups were found for the 6MWT (F=10.75; p<0.001), but no differences were found for the MIP and MEP measures (0.92<F<2.13; 0.13<p<0.40). Significant, negative, and fair correlations were observed between the RDS of the hip and knee muscles and the 6MWT (0.30<r<-0.43; p<0.01) and the HAP (-0.28<r<-0.41; p<0.01). Moderate to good correlations were found between the 6MWT and the HAP (r=0.50; p<0.0001). There were no significant correlations between measures of respiratory strength and any of the investigated variables (-0.11<r<0.12; 0.26<p<0.56). Lower strength deficits and higher functional capacity were associated with higher physical activity levels. However, the moderately active and active groups demonstrated similar strength deficits.


Alcohol consumption and metabolic syndrome: Clinical and epidemiological impact on liver disease.
Alcohol use and metabolic syndrome are highly prevalent in the population and frequently co-exist. Both are implicated in a large range of health problems, including chronic liver disease, hepatocellular carcinoma, and liver-related outcomes (i.e. decompensation or liver transplantation). Studies have yielded mixed results regarding the effects of mild-moderate alcohol consumption on the risk of metabolic syndrome and fatty liver disease, possibly due to methodological differences. The few available prospective studies have indicated that mild-moderate alcohol use is associated with an increase in liver-related outcomes. This conclusion was substantiated by systems biology analyses suggesting that alcohol and metabolic syndrome may play a similar role in fatty liver disease, potentiating an already existing dysregulation of common vital homeostatic pathways. Alcohol and metabolic factors are independently and jointly associated with liver-related outcomes. Indeed, metabolic syndrome increases the risk of liver-related outcomes, regardless of alcohol intake. Moreover, the components of metabolic syndrome appear to have additive effects when it comes to the risk of liver-related outcomes. A number of population studies have implied that measures of central/abdominal obesity, such as the waist-to-hip ratio, can predict liver-related outcomes more accurately than BMI, including in individuals who consume harmful quantities of alcohol. Many studies even point to synergistic interactions between harmful alcohol use and many metabolic components. This accumulating evidence showing independent, combined, and modifying effects of alcohol and metabolic factors on the onset and progression of chronic liver disease highlights the multifactorial background of liver disease in the population. The available evidence suggests that more holistic approaches could be useful for risk prediction, diagnostics and treatment planning.


Mass diffusion and Soret coefficient measurements of triethylene glycol/water binary mixtures by dynamic shadowgraphy.
The investigation of the transport properties of binary fluid mixtures remains a topic of interest in relation to the more challenging studies of ternary mixtures. In fact, the study of the phase boundary limits of the Gibbs composition triangle can be the initial step for a more complete analysis of ternary mixtures. In this paper, we apply the dynamic shadowgraphy optical technique to study non-equilibrium fluctuations induced by the presence of a gradient of temperature and/or concentration in the triethylene glycol (TEG)/water system. These thermodiffusion and free-diffusion experiments aim at measuring the transport properties of samples of the studied system at different experimental conditions. We scan both the average temperature and the TEG concentration, which allows us investigating both positive and negative thermodiffusive behaviours. The obtained values of mass diffusion coefficient are consistent with data available in the literature in the range of temperature investigated in this study. The mass diffusion coefficient of the sample prepared at 0.7 w/w TEG concentration are characterised by shadowgraphy following the two proposed methods, exhibiting consistent results. An increase of the mass diffusion coefficient as a function of the average temperature is highlighted. On the other hand, the thermodiffusion coefficient appears to be independent of the average temperature of the sample at 0.3 w/w TEG concentration.


Nanoscale Mapping of Morphology of Organic Thin Films.
We determine precise nanoscale information about the morphologies of several organic thin film structures using Fourier plane imaging microscopy (FIM). We used FIM microscopy to detect the orientation of molecular transition dipole moments from an extremely low density of luminescent dye molecules, which we call "morphology sensors". The orientation of the sensor molecules is driven by the local film structure and thus can be used to determine details of the host morphology without influencing it. We use symmetric planar phosphorescent dye molecules as the sensors that are deposited into the bulk of organic film hosts during the growth. We demonstrate morphological mapping with a depth resolution to a few Ångstroms that is limited by the ability to determine thickness during deposition, along with an in-plane resolution limited by optical diffraction. Furthermore, we monitor morphological changes arising from thermal annealing of metastable organic films that are commonly employed in photonic devices.


High sensitivity image intensifier-TV detector for x-ray diffraction studies.
A sensitive, efficient image intensifier-TV x-ray detector is described that has been optimized for a large class of diffraction studies of biological structures. All of the major components are commercially available. The system is well suited to measuring the intensity of diffraction patterns that are weak, or changing with time. Because there is no count rate limitation, it is particularly well suited for studies utilizing the high fluxes of synchrotron sources.


Bio-Motivated Two-Level Event-Triggered Controller for Nonlinear Systems.
In this article, a biologically motivated two-level event-triggered mechanism is proposed to design a neuroadaptive controller with exponential convergence property. Specifically, an exponential adaptive neural network controller is designed, and a two-level event-triggered mechanism is developed for a class of nonlinear systems. The two-level event-triggered mechanism, which incorporates both static and dynamic event-triggered features, is motivated by the biological response to low- and high-speed changes in the environment. We also introduce a method in which time-varying control gain is used to achieve exponential convergence of the plant state. The effectiveness of the proposed control scheme is validated by numerical simulations. The minimal interevent time internal is lower bounded by a positive number, so no Zeno behavior occurs.


Free anterolateral thigh adipofascial flap for hemifacial atrophy.
Though the etiology of hemifacial atrophy is still unknown, the aim of its reconstruction is cosmetic amelioration of the defect. Among many techniques introduced, microsurgical reconstruction has become the gold standard to restore the symmetry of the face. Seven free anterolateral thigh adipofascial flaps were used to reconstruct hemifacial atrophy between September 1999 and May 2005. There were 1 male and 6 females in the series. All the patients had unilateral disease (6 right and 1 left). The average age of onset of disease was 11.9 years. The average duration of atrophy was 6.7 years. The patients' average age was 25 years (range, 21 to 32 years), and the average follow-up period was 12.6 months (range, 6 to 36). The size of the transferred flap ranged from 8 x 6 cm to 12 x 13 cm. All adipofascial flaps survived completely. No complications occurred. The result is stable, and no recurrence of facial atrophy was seen in the patients after follow-up. One patient received minor revision because of the bulky reconstructed face. The esthetic appearance of the reconstructed face was acceptable. Despite a variable vascular anatomy that can give rise to some surgical challenge in raising the anterolateral thigh adipofascial flap, the authors conclude that this is a safe and reliable adipofascial flap for hemifacial atrophy.


Fast spin echo vs conventional spin echo in cervical spine imaging.
The major attraction of fast-spin-echo (FSE) imaging is reduced acquisition time; however, careful review of the literature reveals many weaknesses: phase-encoded blurring, truncation artefact, bright fat signal, reduced magnetic susceptibility and increased motion artefact. Our aim was a prospective, blinded comparison of FSE and conventional spin echo (CSE) in the cervical spine. Both sequences were performed in 43 patients (19 males and 24 females; mean age 45 years, range 15-66 years). Twenty-eight patients were studied at 1.5 T and 15 at 0.5 T. Typical sequence parameters were: at 1.5 T, TR/TE 2000/90 CSE and 3000/120 FSE, and at 0.5 T, 2200/80 CSE and 2800/120 FSE. Time saved on the FSE was used to increase the matrix and the number of acquisitions. Two neuroradiologists evaluated the images for pathology, artefacts, disc signal intensity, thecal sac compression and image quality. Ten patients had cord lesions; 2 (20 %) were missed on CSE. In 4 of 10 patients with moderate/severe thecal sac compression, the degree of stenosis was apparently exaggerated on CSE. The mean degree of confidence for the CSE sequences was 1.8 and for the FSE 1.1, where 1 is optimal. For cervical spine imaging, FSE should be preferred to CSE.


The LuxS/AI-2 Quorum-Sensing System Regulates the Algicidal Activity of Shewanella xiamenensis Lzh-2.
Cyanobacterial blooming is an increasing environmental issue all over the world. Algicidal bacteria are potential tools for the control of algal blooms. The algicidal activity in many bacteria exhibits quorum-sensing (QS) dynamics and the regulatory mechanism of this activity in these bacteria is unclear. In this study, combining genomic sequencing and genome editing, we have identified that the primary quorum-sensing system in the isolated algicidal strain Shewanella xiamenensis Lzh-2 is the LuxS/AI-2 signaling pathway. Disruption of the QS system through recombination deletion of the LuxS gene led to a loss of algicides production and algicidal activity. Restoration of the LuxS gene in the deletion mutant compensated the QS system and recovered the algicidal activity. Consequently, we proved that Lzh-2 regulates the algicidal activity through LuxS/AI-2 quorum-sensing system.


Development and Characterization of a Glimepiride-Loaded Gelatin-Coated Mesoporous Hollow Silica Nanoparticle Formulation and Evaluation of Its Hypoglycemic Effect on Type-2 Diabetes Model Rats.
In this study, we prepared gelatin-coated mesoporous hollow silica nanospheres (GSN) as a drug carrier to improve the water solubility and regulate the release rate of glimepiride (GLM). GLM was loaded into GSN by an absorption method, and drug-loaded samples (GLM-GSN) were characterized by differential scanning calorimeter (DSC) and X-ray diffraction (XRD). Cellular uptake and in vivo intestinal uptake experiments were performed in rats. In addition, the studies of in-vitro drug dissolution, pharmacokinetics, and pharmacodynamic experiments also were performed. GLM-GSN showed excellent drug loading (39.7% ± 0.7%) and sustained GLM release. The state of GLM in GSN was amorphous according to DSC and XRD results. Cellular uptake and in vivo intestinal uptake experiments indicated that GSN could be effectively absorbed, and an MTT experiment demonstrated that GSN had good biocompatibility. Furthermore, the GLM-GSN had a higher bioavailability in pharmacokinetics experiments and a prominent hypoglycemic effect on type-2 diabetes model rats in pharmacodynamic experiments. This study clearly shows that GSN is a promising platform for delivering GLM for the treatment of type-2 diabetes.


Experts on age estimation.
The aim of this investigation was to study the biases and accuracy in age estimation of persons selling alcohol. Two experiments are reported, both suggesting that the accuracy in age estimation of Swedish alcohol salespersons is higher than that of control persons. This expertise in age estimation is probably the result of the extensive training Swedish alcohol salespersons go through as a natural part of their profession. Nonetheless, their estimates were not free from bias. Salespersons overestimated the age of target persons below 20 years of age and thus too young to buy alcohol. The results also revealed that controls, in contrast to salespersons, assimilated their estimates towards their own age (i.e. an own-anchor effect). Furthermore, female participants were shown to estimate the age of old target persons (56-65 years) more accurately than male participants. These results are discussed in relation to previous findings on training in age estimation and present jurisdiction.


Ligand-induced acceleration of the intramolecular [3 + 2] cycloaddition between alkynes and alkylidenecyclopropanes.
[chemical reaction: see text]. Bulky phosphite L6 and several sterically robust phosphoramidites are excellent ligands for promoting the Pd-catalyzed [3 + 2] intramolecular cycloaddition between alkylidenecyclopropanes and alkynes. The use of these ligands allows for low catalyst loadings and facilitates the otherwise sluggish cycloaddition of hept-6-ynylidenecyclopropane systems.


[Muckle-Wells syndrome: a rare periodic fever syndrome].
A 41-year-old patient was referred to our rheumatology ward due to a long history of urticaria, joint pain and fever. These symptoms appeared during the evening and then resolved during the night. Extensive testing in the past failed to provide a diagnosis and treatment with high-dose corticosteroids, methotrexate and colchicine was ineffective. Based on clinical criteria, including bilateral sensorineural hearing loss, we diagnosed Muckle-Wells syndrome, a rare auto-inflammatory disease considered one of the hereditary periodic fever syndromes and caused by a mutation in the CIAS1 gene. There was a remarkable response to anakinra, an interleukin-1 receptor antagonist. The favourable results suggest that interleukin-1 plays an important role in the development of this syndrome.


The prognostic value of tumour-stroma ratio in triple-negative breast cancer.
Triple-negative cancer constitutes one of the most challenging groups of breast cancer given its aggressive clinical behaviour, poor outcome and lack of targeted therapy. Until now, profiling techniques have not been able to distinguish between patients with a good and poor outcome. Recent studies on tumour-stroma, found it to play an important role in tumour growth and progression. To evaluate the prognostic value of the tumour-stroma ratio (TSR) in triple-negative breast cancer. One hundred twenty four consecutive triple-negative breast cancer patients treated in our hospital were selected and evaluated. For each patient the Haematoxylin-Eosin (H&E) stained histological sections were evaluated for percentage of stroma. Patients with less than 50% stroma were classified as stroma-low and patients with ≥ 50% stroma were classified as stroma-high. Of 124 triple-negative breast cancer patients, 40% had a stroma-high and 60% had a stroma-low tumour. TSR was assessed by two investigators (kappa 0.74). The 5-years relapse-free period (RFP) and overall survival (OS) were 85% and 89% in the stroma-low and 45% and 65% in the stroma-high group. In a multivariate cox-regression analysis, stroma amount remained an independent prognostic variable for RFP (HR 2.39; 95% CI 1.07-5.29; p = 0.033) and OS (HR 3.00; 95% CI 1.08-8.32; 0.034). TSR is a strong independent prognostic variable in triple-negative breast cancer. It is simple to determine, reproducible and can be easily incorporated into routine histological examination. This parameter can help optimize risk stratification and might lead to future targeted therapies.


Antiemetics for acute gastroenteritis in children.
The alleviation of vomiting due to gastroenteritis in children can help promote oral rehydration and prevent medical visits for dehydration. Recent randomized, placebo-controlled trials of newer antiemetics, such as serotonin antagonists like ondansetron, have demonstrated efficacy in relief of vomiting, decreased need for intravenous fluid therapy and hospitalizations, without serious adverse effects. The ability to arrest further vomiting and prevent intravenous fluid therapy and hospitalization aids children with vomiting so they may tolerate oral fluids and be discharged from medical care. This can reduce the medical care burdens and enhance patient satisfaction. This new pharmaceutical therapy may change how gastroenteritis is managed as it is studied further.


Feature Mining and Health Assessment for Gearboxes Using Run-Up/Coast-Down Signals.
Vibration signals measured in the run-up/coast-down (R/C) processes usually carry rich information about the health status of machinery. However, a major challenge in R/C signals analysis lies in how to exploit more diagnostic information, and how this information could be properly integrated to achieve a more reliable maintenance decision. Aiming at this problem, a framework of R/C signals analysis is presented for the health assessment of gearbox. In the proposed methodology, we first investigate the data preprocessing and feature selection issues for R/C signals. Based on that, a sparsity-guided feature enhancement scheme is then proposed to extract the weak phase jitter associated with gear defect. In order for an effective feature mining and integration under R/C, a generalized phase demodulation technique is further established to reveal the evolution of modulation feature with operating speed and rotation angle. The experimental results indicate that the proposed methodology could not only detect the presence of gear damage, but also offer a novel insight into the dynamic behavior of gearbox.


Detection and identification of allergens from Canadian mustard varieties of Sinapis alba and Brassica juncea.
Currently, information on the allergens profiles of different mustard varieties is rather scarce. Therefore, the objective of this study was to assess protein profiles and immunoglobulin E (IgE)-binding patterns of selected Canadian mustard varieties. Optimization of a non-denaturing protein extraction from the seeds of selected mustard varieties was first undertaken, and the various extracts were quantitatively and qualitatively analyzed by means of protein recovery determination and protein profiling. The IgE-binding patterns of selected mustard seeds extracts were assessed by immunoblotting using sera from mustard sensitized and allergic individuals. In addition to the known mustard allergens-Sin a 2 (11S globulins), Sin a 1, and Bra j 1 (2S albumins)-the presence of other new IgE-binding protein bands was revealed from both Sinapis alba and Brassica juncea varieties. Mass spectrometry (MS) analysis of the in-gel digested IgE-reactive bands identified the unknown ones as being oleosin, β-glucosidase, enolase, and glutathione-S transferase proteins. A bioinformatic comparison of the amino acid sequence of the new IgE-binding mustard proteins with those of know allergens revealed a number of strong homologies that are highly relevant for potential allergic cross-reactivity. Moreover, it was found that Sin a 1, Bra j 1, and cruciferin polypeptides exhibited a stronger IgE reactivity under non-reducing conditions in comparison to reducing conditions, demonstrating the recognition of conformational epitopes. These results further support the utilization of non-denaturing extraction and analysis conditions, as denaturing conditions may lead to failure in the detection of important immunoreactive epitopes.


FACTORS ASSOCIATED WITH COMPLEX REGIONAL PAIN SYNDROME IN SURGICALLY TREATED DISTAL RADIUS FRACTURE.
The aim of this study was to identify factors associated with developing complex regional pain syndrome (CRPS) after surgical treatment for distal radius fracture (DRF). This case-control study analyzed patients seen from January 2014 to January 2016. Results: In our sample of 249 patients, 4% developed CRPS. Associated factors were economic compensation via work disability (odds ratio [OR] 14.3), age (OR 9.38), associated fracture (OR 12.94), and level of impact (OR 6.46), as well as psychiatric history (OR 7.21). Economically-productive aged patients with a history of high-impact trauma and patients with a history of psychiatric disorders have greater risk of developing CRPS after DRF. Level of Evidence III, Case-Control Study.


Differential cytotoxic effects of sodium meta-arsenite on human cancer cells, dental papilla stem cells and somatic cells correlate with telomeric properties and gene expression.
We investigated the effects of sodium meta-arsenite (NaAsO(2)) on human cancer cells (MDA-MB-231, MCF-7 and U-87 MG), dental papilla tissue stem cells (DPSCs) and somatic cells [MRC-5 fetal fibroblasts and adult muscle cells (MCs)] by examining telomeric properties, endogenous reverse transcriptase (RT) activity and the expression of tumorigenesis-linked genes. Half maximal inhibitory concentration (IC(50)) values were higher in DPSCs and MCs, possessing longer telomere lengths when compared to cancer cells. Levels of telomerase and RT activity, and the expression of protein 53 (p53), B-cell lymphoma 2 (BCL2), nuclear factor kappa-light-chain-enhancer of activated B-cells (NFκB), transforming growth factor beta (TGFβ) and vascular endothelial growth factor (VEGF) were significantly lower in cancer cells following sodium meta-arsenite treatment, whereas the effect was absent or marginally detected in DPSCs and somatic cells. Collectively, sodium meta-arsenite effectively induced cellular cytotoxicity by inhibiting telomerase and RT activity, and down-regulating transcript levels in cancer cells with shorter telomere lengths, whereas more tolerance was evident in DPSCs and somatic cells possessing longer telomere lengths.


Neuroophthalmologic manifestations of systemic disease.
Significant contributions on the neuroophthalmologic manifestations of systemic disease have been published in the past year. The first part of this paper is devoted to practical guidelines that may help in the diagnosis and the management of oculomotor disorders, especially in connection with systemic diseases; the second part is focused on specific entities. Including idiopathic intracranial hypertension and ophthalmoplegic migraine in this review might be debatable; however, although the cause of these conditions is still unclear, relevant findings suggest that they may be the consequence of other adjacent processes rather than the cause itself. We believe that the diagnosis of idiopathic intracranial hypertension as well as that of ophthalmoplegic migraine remains a challenging issue for the ophthalmologist and may easily be overlooked.


The Role of Sense of Voice Presence and Anxiety Reduction in AVATAR Therapy.
AVATAR therapy offers a unique therapeutic context that uses virtual reality technology to create a virtual embodiment of the voice-hearing experience, enabling the person to visualize their persecutory voice and engage in real-time "face-to-face" dialogue. The present study explores, for the first time, the contribution of sense of voice presence, together with session-by-session reduction of anxiety and paranoid attributions about the avatar, to changes in primary outcomes following AVATAR therapy. Data from 39 participants, who completed AVATAR therapy and attended a 12-week follow-up assessment, were analysed. Mid- to high-levels of sense of voice presence were reported across the therapy sessions, along with significant reductions of anxiety levels and paranoid attributions about the avatar. The interaction of sense of voice presence and reduction of anxiety was associated with two of the significant therapy outcomes: PSYRATS total and frequency of voices. The findings suggest that improvements in voice severity and frequency at post AVATAR therapy may be influenced by the combination of feeling less anxious in the context of a realistic simulation of the voice, while voice-related distress may involve additional cognitive and relational processes.


Role of antioxidants in the treatment of male infertility.
Male infertility continues to be a clinical challenge of increasing significance. While male factors such as decreased semen quality are responsible for 25% of all infertility issues, the etiology of suboptimal semen quality is poorly understood. Many physiological, environmental, and genetic factors have been implicated, including oxidative stress. Oxidative stress is induced by reactive oxygen species (ROS), or free radicals, and although ROS are required for critical aspects of sperm function, excessive levels of ROS can negatively impact sperm quality. The origin of ROS generation, and the etiologies of increased ROS in men with suboptimal sperm quality have only recently been elucidated, offering multiple targets for potential therapy. Here, we present a critical review of the literature describing the role of oxidative stress on decreased sperm function, as well as the role of antioxidants in the treatment of male factor infertility.


Peroneal nerve injury with foot drop complicating ankle sprain--a series of four cases with review of the literature.
Foot drop has many etiologies. One rarely mentioned and often neglected reason for foot drop is an acute inversion sprain of the ankle. Over the past 14 years, a collection of 32 cases of foot drop have been compiled in our orthopaedic and physiatric practices. All cases had appropriate evaluations, including electrodiagnostic studies (electromyography and nerve conduction studies) to determine the location and type of injury. Treatment and follow-up are also discussed. Of the 32 case studies, four were caused by a straightforward acute inversion sprain of the ankle. These cases are described with the electrodiagnostic evaluations, treatments, and outcomes. Proposed mechanisms for this type of foot drop are discussed, including traction and compression of the common peroneal nerve as it winds around the neck of the fibula, and possible compression by hematoma. Surgical versus conservative treatment is described. The functional impairment associated with foot drop is detailed.


Heterogeneity of tumor cells in terms of cancer-initiating cells.
Tumors derive from a single cell clone but consist of heterogeneous cell subpopulations with diverse features and functions. A limited number of subclones with a selective advantage can initiate tumors when inoculated into immunocompromised mice, and are called cancer-initiating cells (CICs). CICs can be isolated from the bulk of tumors on the basis of their characteristics, such as high reagent efflux, degradation of reactive oxygen species, and aldehyde dehydrogenase (ALDH) activity. Under normal conditions, new CICs are produced by existing CICs rather than non-CICs. However, under stress conditions, non-CICs can occasionally produce CICs, a phenomenon known as plasticity. The dynamic exchange between CICs and non-CICs may enable tumors to survive under unfavorable conditions. CICs are located in a small portion of tumors. This suggests that microenvironmental factors induce or inhibit the CIC phenotype, which might be regulated by intercellular signaling between tumor cells. This review describes isolation of CICs from tumor cell populations and the microenvironmental factors that regulate CIC phenotypes in uterine cancer and lymphoma.


Altered islet prohormone processing: a cause or consequence of diabetes?
Peptide hormones are first produced as larger precursor prohormones that require endoproteolytic cleavage to liberate the mature hormones. A structurally conserved but functionally distinct family of nine prohormone convertase enzymes (PCs) are responsible for cleavage of protein precursors, of which PC1/3 and PC2 are known to be exclusive to neuroendocrine cells and responsible for prohormone cleavage. Differential expression of PCs within tissues defines prohormone processing; whereas glucagon is the major product liberated from proglucagon via PC2 in pancreatic α-cells, proglucagon is preferentially processed by PC1/3 in intestinal L cells to produce glucagon-like peptides 1 and 2 (GLP-1, GLP-2). Beyond our understanding of processing of islet prohormones in healthy islets, there is convincing evidence that proinsulin, pro-islet amyloid polypeptide (proIAPP), and proglucagon processing is altered during prediabetes and diabetes. There is predictive value of elevated circulating proinsulin or proinsulin-to-C-peptide ratio for progression to type 2 diabetes, and elevated proinsulin or proinsulin-to-C-peptide ratio is predictive for development of type 1 diabetes in at-risk groups. After onset of diabetes, patients have elevated circulating proinsulin and proIAPP, and proinsulin may be an autoantigen in type 1 diabetes. Furthermore, preclinical studies reveal that α-cells have altered proglucagon processing during diabetes, leading to increased GLP-1 production. We conclude that despite strong associative data, current evidence is inconclusive on the potential causal role of impaired prohormone processing in diabetes and suggest that future work should focus on resolving the question of whether altered prohormone processing is a causal driver or merely a consequence of diabetes pathology.


Economic contribution and attitude towards alien freshwater ornamental fishes of pet store owners in Klang Valley, Malaysia.
Malaysia is one of the top ten countries in the world that produce freshwater ornamental fishes. This industry can offer better livelihood opportunities to many poor households. However, most of the produced ornamental fishes are alien to Malaysia. In this study, we explore the contribution of alien freshwater fishes to the income of ornamental fish store owners and their attitude towards alien freshwater fishes within Klang Valley, Malaysia. Using a structured questionnaire, we surveyed 70 pet stores out of which 54 (81.42%) store owners responded. Most of the pet store owners were male (72%), Chinese (83%), and the highest educational level was at the secondary level (79%). Most of the pet store owners reported a monthly income of RM 2001-RM 5000 (78%) and were married (73%). Using Chi-square (χ2) test, significant relationships (p < 0.05) existed between the attitude of store owners towards alien ornamental fish species versus educational level (χ2 = 16.424, p = 0.007) and contribution of alien ornamental fishes to the pet store owners' income (χ2 = 27.266, p = 0.003). Fish sales as the main income source also related significantly with the impact of fish selling business on income level (χ2 = 10.448, p = 0.007). This study showed that the ornamental fish sales contributed over half of the income (51-100%) from the businesses of store owners. Almost half of the respondents (42%) reported that alien ornamental fish was the highest contributor to their income from the ornamental fish sale. While the mismanagement of alien ornamental fishes could give various negative ecological impacts, the socio-economic benefits of these fishes cannot be denied.


Dorsal Raphe 5-HT Neurons Utilize, But Do Not Generate, Negative Aversive Prediction Errors.
The dorsal raphe nucleus (DRN) contains the largest population of serotonin (5-HT) neurons in the central nervous system. 5-HT, synthesized via tryptophan hydroxylase 2 (Tph2), is a widely functioning neuromodulator implicated in fear learning. Here, we sought to investigate whether DRN 5-HT is necessary to reduce fear via negative prediction error (-PE). Using male and female TPH2-cre rats, DRNtph2+ cells were selectively deleted via cre-caspase (rAAV5-Flex-taCasp3-TEVp) in experiment 1. Rats then underwent fear discrimination during which three cues were associated with unique foot shock probabilities: safety p = 0.00, uncertainty p = 0.375, and danger p = 1.00. Rats then received selective extinction to the uncertainty cue, a behavioral manipulation designed to probe -PE. Deleting DRNtph2+ cells had no impact on initial discrimination but slowed selective extinction. In experiment 2, we used a within-subjects optogenetic inhibition design to causally implicate DRNtph2+ cells in prediction error signaling. Male and female TPH2-cre rats received intra-DRN infusions of cre-dependent halorhodopsin (rAAV5-Ef1a-DIO-eNpHR3.0-eYFP) or cre-YFP. DRNtph2+ cells were inhibited specifically during the time of prediction error or a control period. Illumination during either positive prediction error (+PE) or control periods had no impact on fear to the uncertainty cue. Inhibition of DRNtph2+ cells at the time of -PE did not impact immediate fear, but facilitated selective extinction in postillumination sessions. Together, these results demonstrate a role for DRNtph2+ cells in using, but not generating, -PE to weaken cue-shock associations.


The effect of adrenal cortex hormones on heart and skeletal muscles amylases activity under conditions of alpha- and beta-adrenergic stimulation.
Influence of hydrocortisone (H), deoxycorticosterone (D), phenylephrine (F) and isoprenaline (I) on activity of alpha amylase (AA) = 4-glucohydrolase alpha-1, 4-glucan [3, 2, 1, 1] and glucoamylase (GA) = glucohydrolase alpha-1, 4-glucan [3, 2, 1, 3] was investigated in heart and skeletal muscle in rats, before and after adrenalectomy (AT). It was found that AT inhibits a rise in the activities of AA and GA caused by F in both investigated muscles, whereas the I given after AT loses its ability to inhibit GA activity in heart and skeletal muscle. Both H and D administered after AT recover the ability of F to an increase in the investigated enzyme activity whereas recovers I ability to inhibit amylase activity in the presence of H. It can be concluded that inhibition of amylase activity is muscles results from interaction between beta-adrenergic stimulation with glucocorticoids, while the increase in activities of these enzymes is a result of interaction between alpha-adrenergic stimulation with mineral tropic action of both D and H.


Phytoremediation of petroleum hydrocarbon-contaminated saline-alkali soil by wild ornamental Iridaceae species.
As a green remediation technology, phytoremediation is becoming one of the most promising methods for treating petroleum hydrocarbons (PHCs)-contaminated soil. Pot culture experiments were conducted in this study to investigate phytoremediation potential of two representative Iridaceae species (Iris dichotoma Pall. and Iris lactea Pall.) in remediation of petroleum hydrocarbon-contaminated saline-alkali soil from the Dagang Oilfield in Tianjin, China. The results showed that I. lactea was more endurable to extremely high concentration of PHCs (about 40,000 mg/kg), with a relatively high degradation rate of 20.68%.The degradation rate of total petroleum hydrocarbons (TPHs) in soils contaminated with 10,000 and 20,000 mg/kg of PHCs was 30.79% and 19.36% by I. dichotoma, and 25.02% and 19.35% by I. lactea, respectively, which improved by 10-60% than the unplanted controls. The presence of I. dichotoma and I. lactea promoted degradation of PHCs fractions, among which saturates were more biodegradable than aromatics. Adaptive specialization was observed within the bacterial community. In conclusion, phytoremediation by I. dichotoma should be limited to soils contaminated with ≤20,000 mg/kg of PHCs, while I. lactea could be effectively applied to phytoremediation of contaminated soils by PHCs with at least 40,000 mg/kg.


The status of infection prevention and control structures in Eastern China based on the IPCAF tool of the World Health Organization.
The burden of healthcare-associated infections (HAIs) and the spread of antimicrobial resistance can be potentially preventable through comprehensive infection prevention and control (IPC) programs. However, information on the current state of IPC implementation is rare in China. A cross-sectional study was conducted in Zhejiang province, China, from April to June 2021. The Zhejiang nosocomial infection control and quality improvement center (NICQI) cooperated with 11 municipal NICQI centers to introduce the purpose of this study and invite all licensed secondary and tertiary hospitals in Zhejiang province through WeChat group. The questionnaire had three sections, including information about participating hospitals, demographic information about IPs, and the Chinese version of the Infection Prevention and Control Assessment Framework that covered eight core components (CC). Of the 382 hospitals invited, 222 (58.1% response rate) accepted and completed the online questionnaire. The overall median score of the participating hospitals was 682 (630-723), which corresponded to an advanced level of IPC. There was a significant difference in scores between hospitals types (P < 0.001). Profound differences were revealed regarding the scores of the individual components, with CC2 (IPC guidelines) and CC6 (Monitoring/audit of IPC practices and feedback) having the highest (100) and lowest (65) median scores, respectively. Only 23 (10.4%) hospitals reported assessing facility safety culture. IPC structures are at a relatively high level in acute care hospitals in Eastern China. The identified potential areas for improvement were similar to those identified in developed countries, particularly regarding multimodal strategies for implementation and safety culture construction. Meanwhile, the Chinese government should pay more attention to IPC resources and practices among secondary care hospitals.


Identification of Natural Killer Cells by Immunoelectron Microscopy
We employed immunoelectron microscopic techniques to localize natural killer cells (NK cells) in human lymph node and tonsil. These tissues were embedded in Lowicryl K4M. Thin sections were first reacted with anti-Leu-7 followed by anti-UCHL. Colloidal gold particles of different sizes were used as a label. NK cells were localized primarily in paracortical T-cell regions. The cells typed with these antisera include both large granular and agranular lymphocytes. No other cell types expressed the NK phenotype. These results illustrate the versatility of immunoelectron microscopy to solve problems beyond the resolution of the light microscope.