Expression of heat shock protein 27 in gut tissue of growing pigs fed diets without and with inclusion of chicory fiber.
The physiological expression of cytoprotective heat shock protein 27 (Hsp27) in the gut was investigated in eighteen 7-wk-old pigs fed one of 3 fiber-rich diets for 18 d. The diets were a cereal-based control diet and a cereal-based diet with inclusion of either 80 g/kg chicory forage (CF80) or chicory root (CR80). Immunohistochemical staining showed that Hsp27 was expressed in all the samples from ileum and colon. The expression was most intensive in the apical intestinal epitheliums in close contact with luminal contents and lighter in crypt cells. The ileal Peyer's patches showed a strong expression of Hsp27, which was highly correlated with Hsp27 expression in the ileal epithelial cells (P = 0.003). The frequency of ileal Hsp27 expression with the most intensive staining was distributed higher in pigs fed chicory forage diet (CF80, 25%) followed by chicory root diet (CR80, 16.7%) and the control (11.1%). In proximal colon, the frequency of expression showed a similar pattern for the different diets. The intestinal microbiota profile was characterized with the intention to find correlations to heat shock protein (Hsp) expression in pig gastrointestinal (GI) tract and showed that the distal ileum and proximal colon encompass its own unique microbial profile. However, no significant relationship was found between gut microbiota diversity and Hsp27 expression. These indicate that Hsp27 expression in the porcine gut could be associated with specific dietary fiber components but not the overall microbiota diversity.


Light and electron microscopic distribution of the AMPA receptor subunit, GluR2, in the spinal cord of control and G86R mutant superoxide dismutase transgenic mice.
Excitotoxicity has been hypothesized to contribute to amyotrophic lateral sclerosis (ALS) neurodegeneration. The similar pattern of vulnerability in the spinal cord of mutant superoxide dismutase (SOD-1) transgenic mice and mice treated with excitotoxins supports a role for excitotoxicity in the mechanism of degeneration. The distribution of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) class of glutamate receptors (GluRs) with different calcium permeabilities has been proposed as an explanation for this differential vulnerability. GluR2 appears to be the dominant determinant of calcium permeability for AMPA receptors; thus, it is critical for their contribution to excitotoxic mechanisms. In this study, we investigate the distribution of GluR2 immunoreactivity in the spinal cord of control and SOD-1 transgenic mice. GluR2 immunoreactivity is present equally within vulnerable neurons (i.e., motor neurons and calretinin-immunoreactive neurons) as well as nonvulnerable neurons (i.e., calbindin-immunoreactive neurons and dorsal horn neurons). In addition, postembedding immunoelectron microscopy reveals that GluR2 is present in synapses of dorsal and ventral horn neurons and that the percentage of labeled synapses and numbers of immunogold particles per synapse do not vary between these spinal cord regions. Comparing control mice with SOD-1 transgenic mice, at both the light and the electron microscopic levels, the distribution and intensity of GluR2-immunoreactivity do not appear to be altered. These results suggest that the cellular and synaptic distribution of GluR2 is not a determinant of the selective vulnerability observed in SOD-1 transgenic mice or in ALS patients.


Trial of Spesolimab for Generalized Pustular Psoriasis.
Generalized pustular psoriasis (GPP) is a rare, life-threatening, inflammatory skin disease characterized by widespread eruption of sterile pustules. Interleukin-36 signaling is involved in the pathogenesis of this disorder. Spesolimab, a humanized anti-interleukin-36 receptor monoclonal antibody, is being studied for the treatment of GPP flares. In a phase 2 trial, we randomly assigned patients with a GPP flare in a 2:1 ratio to receive a single 900-mg intravenous dose of spesolimab or placebo. Patients in both groups could receive an open-label dose of spesolimab on day 8, an open-label dose of spesolimab as a rescue medication after day 8, or both and were followed to week 12. The primary end point was a Generalized Pustular Psoriasis Physician Global Assessment (GPPGA) pustulation subscore of 0 (range, 0 [no visible pustules] to 4 [severe pustulation]) at the end of week 1. The key secondary end point was a GPPGA total score of 0 or 1 (clear or almost clear skin) at the end of week 1; scores range from 0 to 4, with higher scores indicating greater disease severity. A total of 53 patients were enrolled: 35 were assigned to receive spesolimab and 18 to receive placebo. At baseline, 46% of the patients in the spesolimab group and 39% of those in the placebo group had a GPPGA pustulation subscore of 3, and 37% and 33%, respectively, had a pustulation subscore of 4. At the end of week 1, a total of 19 of 35 patients (54%) in the spesolimab group had a pustulation subscore of 0, as compared with 1 of 18 patients (6%) in the placebo group (difference, 49 percentage points; 95% confidence interval [CI], 21 to 67; P<0.001). A total of 15 of 35 patients (43%) had a GPPGA total score of 0 or 1, as compared with 2 of 18 patients (11%) in the placebo group (difference, 32 percentage points; 95% CI, 2 to 53; P = 0.02). Drug reactions were reported in 2 patients who received spesolimab, in 1 of them concurrently with a drug-induced hepatic injury. Among patients assigned to the spesolimab group, infections occurred in 6 of 35 (17%) through the first week; among patients who received spesolimab at any time in the trial, infections had occurred in 24 of 51 (47%) at week 12. Antidrug antibodies were detected in 23 of 50 patients (46%) who received at least one dose of spesolimab. In a phase 2 randomized trial involving patients with GPP, the interleukin-36 receptor inhibitor spesolimab resulted in a higher incidence of lesion clearance at 1 week than placebo but was associated with infections and systemic drug reactions. Longer and larger trials are warranted to determine the effect and risks of spesolimab in patients with pustular psoriasis. (Funded by Boehringer Ingelheim; Effisayil 1 ClinicalTrials.gov number, NCT03782792.).


Simulation Training Improves Trainee Technical Skill and Procedural Attitudes in Endoscopic Gastrostomy Tube Placement.
Percutaneous endoscopic gastrostomy (PEG) tube placement remains a core competency of gastroenterology fellowship, although this procedure is performed infrequently. Some training programs lack sufficient procedural volume for trainees to develop confidence and competence in this procedure. We aimed to determine the impact of a simulation-based educational intervention on trainee technical skill and procedural attitudes in simulated PEG tube placement. Gastroenterology fellows were invited to participate in the study. Baseline procedural attitudes toward PEG tube placement (self-confidence, perceived skill level, perceived level of required supervision) were assessed before simulation training using a Likert scale. Baseline technical skills were assessed by video recording-simulated PEG tube placement on a PEG tube simulator with scoring using a procedural checklist. Fellows next underwent individualized simulation training and repeated simulated PEG tube placement until greater than 90% of checklist items were achieved. Procedural attitudes were reassessed directly after the simulation. Technical skill and procedural attitudes were then reassessed 6 to 12 weeks later (delayed posttraining). Twelve fellows completed the study. Simulation training led to significant improvement in technical skill at delayed reassessment (52.9 ± 14.3% vs. 78.0 ± 8.9% correct, P = 0.0002). Simulation training also led to significant immediate improvements in self-confidence (2.1 ± 0.7 vs. 3.1 ± 0.3, P = 0.001), perceived skill level (2.2 ± 1.0 vs. 4 ± 1.1, P < 0.001), and perceived level of required supervision (2.2 ± 0.9 vs. 3.2 ± 0.6, P = 0.003). Simulation training led to sustained improvements in gastroenterology fellows' technical skill and procedural attitudes in PEG tube placement. Incorporation of simulation curricula in gastroenterology fellowships for this infrequently performed procedure should be considered.


iTRAQ-based quantitative proteomic analysis of two transgenic soybean lines and the corresponding non-genetically modified isogenic variety.
To investigate the unintended effects of genetically modified (GM) crops, an isobaric tags for relative and absolute quantitation (iTRAQ)-based comparative proteomic analysis was performed with seed cotyledons of two GM soybean lines, MON87705 and MON87701×MON89788, and the corresponding non-transgenic isogenic variety A3525. Thirty-five differentially abundant proteins (DAPs) were identified in MON87705/A3525, 27 of which were upregulated and 8 downregulated. Thirty-eight DAPs were identified from the MON87701×MON89788/A3525 sample, including 29 upregulated proteins and 9 downregulated proteins. Pathway analysis showed that most of these DAPs participate in protein processing in endoplasmic reticulum and in metabolic pathways. Protein-protein interaction analysis of these DAPs demonstrated that the main interacting proteins are associated with post-translational modification, protein turnover, chaperones and signal transduction mechanisms. Nevertheless, these DAPs were not identified as new unintended toxins or allergens and only showed changes in abundance. All these results suggest that the seed cotyledon proteomic profiles of the two GM soybean lines studied were not dramatically altered compared with that of their natural isogenic control.


Morphine and ethanol pretreatment effects on expression and extinction of ethanol-induced conditioned place preference and aversion in mice.
Opioid receptor antagonists reliably alter the expression or extinction of ethanol's conditioned motivational effects as indexed by the place conditioning procedure, suggesting endogenous opioids are normally involved. These studies examined how exogenous stimulation of opioid receptors alters ethanol's conditioned rewarding and aversive effects. Drugs that either directly (morphine) or indirectly (ethanol) stimulate opioid receptors were tested for their effects on the expression and extinction of ethanol-induced conditioned place preference (CPP) and conditioned place aversion (CPA). Male DBA/2J mice were exposed to unbiased ethanol (2 g/kg) conditioning procedures that produced either CPP (experiments 1-2) or CPA (experiments 3-4). Morphine (0, 2.5, 5, or 10 mg/kg) was injected before three post-conditioning tests in experiments 1 and 3, whereas ethanol (0, 1, 2, or 3 g/kg) was injected before tests in experiments 2 and 4. All groups received vehicle on test 4 to determine whether the drug pretreatments altered the course of extinction. Morphine dose-dependently enhanced CPP expression (experiment 1), but ethanol dose-dependently reduced CPP expression (experiment 2). Test 4 showed no differences between drug-treated mice and mice given vehicle on all tests. Morphine had no effect on expression or extinction of ethanol-induced CPA (experiment 3). The highest ethanol dose (3 g/kg) interfered with CPA expression, but not extinction (experiment 4). Pretreatment drug effects on ethanol CPP and CPA expression were most likely a byproduct of their activity altering effects rather than opioid-receptor mediated modulation of ethanol's conditioned motivational effects. Neither drug affected the course of extinction.


Associations among co-occurring medical conditions and cognition, language, and behavior in Down syndrome.
Specific medical conditions are more prevalent in Down syndrome (DS) compared to the general population. Medical heterogeneity has also been hypothesized to contribute to variability in outcomes in DS. This project aimed to examine the association between medical conditions (i.e., gastrointestinal issues, hearing loss, vision problems, and congenital heart defects) and cognition, language, and behavior in children and adolescents with DS. Participants were 73 children and adolescents with DS, ages 6-17 years (M = 12.67, SD = 3.16). Caregivers reported on participants' medical conditions, social behaviors, maladaptive behaviors, and executive function. Child cognitive abilities were also assessed. Of the 73 participants, 34.2% had gastrointestinal issues, 12.3% had uncorrected hearing loss, 26.0% had uncorrected vision problems, and 31.5% had congenital heart defects. Participants with gastrointestinal issues had significantly more challenges with social behaviors, maladaptive behaviors, and executive function compared to those without gastrointestinal issues. The associations identified between gastrointestinal issues and caregiver-reported behavioral characteristics in youth with DS contributes to our understanding of the interrelation between co-occurring medical conditions and child outcomes and has implications for approaches to care for individuals with DS.


[Do people exercise less than they think?].
Physical activity is important for health. Physical inactivity is an independent risk factor for disease and speeds up aging. People say in health surveys that they exercise more than they did before. This does not conform with two important facts: Over the last 20 years people have reduced their calorie intake, but their weight is increasing. The only possible explanation is that they exercise less than before, probably because their daily life requires less and less physical activity. We believe this to be a serious health problem, and support the most recent recommendation: A daily walk for 30 minutes.


Comparison of action levels for patient-specific quality assurance of intensity modulated radiation therapy and volumetric modulated arc therapy treatments.
To perform a comprehensive and systematic comparison of fixed-beam IMRT and volumetric modulated arc therapy (VMAT) patient-specific QA measurements for a common set of geometries using typical measurement methods. Fixed-beam IMRT and VMAT plans were constructed for structure set geometries provided by AAPM Task Group 119. The plans were repeatedly delivered across multiple measurement sessions, and the resulting dose distributions were measured with (1) radiochromic film and ionization chamber and (2) a commercial two-dimensional diode array. The resulting QA measurements from each delivery technique were then analyzed, compared, and tested for statistically significant differences. Although differences were noted between QA results for some plans, neither modality showed consistently better agreement of measured and planned doses: of the 22 comparisons, IMRT showed better QA results in 11 cases, and VMAT showed better QA results in 11 cases. No statistically significant differences (p < 0.05) between IMRT and VMAT QA results were found for point doses measured with an ionization chamber, planar doses measured with radiochromic film, or planar doses measured with a two-dimensional diode array. These results suggest that it is appropriate to apply patient-specific QA action levels derived from fixed-beam IMRT to VMAT.


The Cytokine and Bone Protein Expression by Ellagic Acid-Hydroxyapatite in Bone Remodelling Model.
Ellagic acid, a phenolic compound with anti-inflammatory potential, can be used to accelerate the bone healing process and affect human health, while hydroxyapatite is the most commonly used bone graft material. Using a combination of the two materials results in reduced inflammation and increased osteogenesis. This study aimed to determine the effects of combining ellagic acid and hydroxyapatite in bone marker remodelling by analysing the expression of tumour necrosis factor-α (TNF-α), interleukin 10 (IL-10), bone morphogenetic 4 protein (BMP-4), and osteopontin (OPN). Thirty Wistar rats were used in the study. A defect was created in each animal's femur using a low-speed diamond bur. In the control group, the bone was then treated with polyethylene glycol (PEG). In one of the other groups, the bone was treated with hydroxyapatite, and in the other, with ellagic acid-hydroxyapatite. The femur was biopsied 7 days after the procedure and again 14 days after the procedure, and an indirect immunohistochemical (IHC) examination was performed for TNF-α, IL-10, BMP-4, and OPN expression. The ellagic acid-hydroxyapatite decreased TNF-α expression in the bone tissue after 7 days and again after 14 days (p < 0.05). On the other hand, it increased IL-10, BMP-4, and OPN expression (p < 0.05) during the same time periods. Ellagic acid-hydroxyapatite plays a role in bone marker remodelling by decreasing the expression of TNF-α and increasing the expression of IL-10, BMP-4, and OPN. This hydroxyapatite combination can therefore be recommended for use as bone graft material.


What Are the Implications of the STAR*D Trial for Primary Care? A Review and Synthesis.
Although results of the Sequenced Treatment Alternatives to Relieve Depression (STAR*D) trial have been widely disseminated to mental health care providers, hitherto, primary care providers, who diagnose and manage most individuals with depressive syndromes, have had minimal exposure to the study's key findings. We aim to provide translational implications of the STAR*D trial for primary care practitioners as well as for future research vistas. A PubMed search was carried out with key search terms STAR*D and treatment-resistant depression found in articles published from 2001 through 2007. Articles reporting on the STAR*D outcomes at each sequence of treatment were the primary sources for review. Results from the primary outcome measures at each sequential treatment were extracted and reviewed. Articles reporting variables affecting the probability of achieving remission were also selected. The STAR*D trial is the largest effectiveness study evaluating next-step therapies in real-world patients with major depressive disorder. The ecological validity of the study results are burnished by several methodological factors, including the enrollment of both publicly and privately insured patients, the recruitment of patients in primary and specialty care settings, the broad inclusion criteria, the use of pharmacologic and psychosocial (i.e., cognitive-behavioral therapy) treatment options, the use of measurement-based care, and the randomized clinical equipoise design. Taken together, remission rates of approximately 50% to 55% were reported after 2 sequential treatment interventions. A substantial percentage of individuals achieving remission do so after 6 weeks of treatment. The probabilities of achieving remission with third- and fourth-step therapy were considerably lower, i.e., ≤ 25%. The probabilities of relapse during continuation therapy increased as a function of number of treatment trials required to achieve remission. There is no evidence that individuals failing to achieve remission with a selective serotonin reuptake inhibitor (SSRI) have a greater probability of remitting with a separate class antidepressant versus an alternative SSRI. A window of therapeutic opportunity appears to exist insofar as acute remission rates in major depressive disorder are greatest with the first 2 sequential treatments. Taken together, measurement-based care affords the greatest probability that an individual will achieve remission. Despite optimal continuation treatment, relapse rates remain significant, underscoring the chronicity of depressive disorders.


The phosphatidylinositol-3-phosphate 5-kinase inhibitor apilimod blocks filoviral entry and infection.
Phosphatidylinositol-3-phosphate 5-kinase (PIKfyve) is a lipid kinase involved in endosome maturation that emerged from a haploid genetic screen as being required for Ebola virus (EBOV) infection. Here we analyzed the effects of apilimod, a PIKfyve inhibitor that was reported to be well tolerated in humans in phase 2 clinical trials, for its effects on entry and infection of EBOV and Marburg virus (MARV). We first found that apilimod blocks infections by EBOV and MARV in Huh 7, Vero E6 and primary human macrophage cells, with notable potency in the macrophages (IC50, 10 nM). We next observed that similar doses of apilimod block EBOV-glycoprotein-virus like particle (VLP) entry and transcription-replication competent VLP infection, suggesting that the primary mode of action of apilimod is as an entry inhibitor, preventing release of the viral genome into the cytoplasm to initiate replication. After providing evidence that the anti-EBOV action of apilimod is via PIKfyve, we showed that it blocks trafficking of EBOV VLPs to endolysosomes containing Niemann-Pick C1 (NPC1), the intracellular receptor for EBOV. Concurrently apilimod caused VLPs to accumulate in early endosome antigen 1-positive endosomes. We did not detect any effects of apilimod on bulk endosome acidification, on the activity of cathepsins B and L, or on cholesterol export from endolysosomes. Hence by antagonizing PIKfyve, apilimod appears to block EBOV trafficking to its site of fusion and entry into the cytoplasm. Given the drug's observed anti-filoviral activity, relatively unexplored mechanism of entry inhibition, and reported tolerability in humans, we propose that apilimod be further explored as part of a therapeutic regimen to treat filoviral infections.


Molecular analysis of bacterial communities associated with the roots of Douglas fir (Pseudotsuga menziesii) colonized by different ectomycorrhizal fungi.
We studied the effect of ectomycorrhizal fungi on bacterial communities colonizing roots of Douglas fir (Pseudotsuga menziesii). Mycorrhizal tips were cleaned of soil and separated based on gross morphological characteristics. Sequencing of the internal transcribed spacers of the nuclear rRNA gene cluster indicated that the majority of the tips were colonized by fungi in the Russulaceae, with the genera Russula and Lactarius comprising 70% of the tips. Because coamplification of organellar 16S rRNA genes can interfere with bacterial community analysis of root tips, we developed and tested a new primer pair that permits amplification of bacterial 16S rRNA genes but discriminates more effectively against organellar sequences than commonly used bacterial primer sets. We then used terminal restriction fragment length polymorphism (T-RFLP) and sequence analysis of the 16S rRNA gene to examine differences in bacterial communities associated with the mycorrhizal tips. Cluster analysis of T-RFLP profiles indicated that there were different bacterial communities among the root tips; however, the communities did not seem to be affected by the taxonomic identity of the ectomycorrhizal fungi. Terminal restriction fragment profiling and sequencing of cloned partial 16S rRNA genes indicated that most bacteria on the ectomycorrhizal tips were related to the Alphaproteobacteria and the Bacteroidetes group.


Naturally occurring autoantibodies to skeletal proteins from human red blood cells.
The IgG fraction of sera of healthy human subjects contains natural antibodies to cytoskeletal elements of the donors own red blood cell membranes. Autoantibodies to spectrin are characterized in more detail: their Fab portion binds to the antigen. Autoantibodies, affinity-purified on immobilized spectrin band 1, precipitate 0.4 microgram of spectrin dimer per 1 microgram of autoantibody. They bind to band 1 and cross-react with band 2 of spectrin as well as with breakdown products of spectrin on blots from separated membrane polypeptides. Autoantibodies purified on spectrin band 2 after absorption on band 1 do not cross-react with band 1. The evidence strongly suggests the existence of such autoantibodies in healthy human subjects. This finding indicates that autoantibody production to normally unexposed antigens is not suppressed in ontogeny. These anti-cytoskeleton autoantibodies may have a physiologic role in clearance of debris from lysed cells. Their existence may open a new understanding of elevated anti-spectrin autoantibody concentrations in diseases with different etiologies.


Ezatiostat hydrochloride for the treatment of myelodysplastic syndromes.
Myelodysplastic syndromes (MDSs) are associated with significant morbidity due to ineffective hematopoiesis. Given the limited number of drugs approved by the FDA, there is a need for new therapeutic options. Ezatiostat is a novel agent targeting oxidative stress via inhibition of glutathione S-transferase 1. Herein, the authors summarize the standard of care in order to build the framework for therapeutic advancements. The purpose of this paper is to review the body of preclinical and clinical research literature on the investigational agent ezatiostat hydrochloride (TLK199) for the treatment of MDSs. The article includes details of the pathophysiology, pharmacology, toxicity and efficacy of ezatiostat hydrochloride from controlled studies in patients with myelodysplasia. MDS clonal heterogeneity and clonal architecture complexity has presented a significant technical challenge in developing effective therapies. Ezatiostat offers a unique and specific mechanism to improve the transfusion burden associated with myelodysplasia. Since it is tolerable as a monotherapy, combining ezatiostat with agents such as lenalidomide may have the most potential benefit.


Early back-to-Africa migration into the Horn of Africa.
Genetic studies have identified substantial non-African admixture in the Horn of Africa (HOA). In the most recent genomic studies, this non-African ancestry has been attributed to admixture with Middle Eastern populations during the last few thousand years. However, mitochondrial and Y chromosome data are suggestive of earlier episodes of admixture. To investigate this further, we generated new genome-wide SNP data for a Yemeni population sample and merged these new data with published genome-wide genetic data from the HOA and a broad selection of surrounding populations. We used multidimensional scaling and ADMIXTURE methods in an exploratory data analysis to develop hypotheses on admixture and population structure in HOA populations. These analyses suggested that there might be distinct, differentiated African and non-African ancestries in the HOA. After partitioning the SNP data into African and non-African origin chromosome segments, we found support for a distinct African (Ethiopic) ancestry and a distinct non-African (Ethio-Somali) ancestry in HOA populations. The African Ethiopic ancestry is tightly restricted to HOA populations and likely represents an autochthonous HOA population. The non-African ancestry in the HOA, which is primarily attributed to a novel Ethio-Somali inferred ancestry component, is significantly differentiated from all neighboring non-African ancestries in North Africa, the Levant, and Arabia. The Ethio-Somali ancestry is found in all admixed HOA ethnic groups, shows little inter-individual variance within these ethnic groups, is estimated to have diverged from all other non-African ancestries by at least 23 ka, and does not carry the unique Arabian lactase persistence allele that arose about 4 ka. Taking into account published mitochondrial, Y chromosome, paleoclimate, and archaeological data, we find that the time of the Ethio-Somali back-to-Africa migration is most likely pre-agricultural.


Invited review: Activation of G proteins by GTP and the mechanism of Gα-catalyzed GTP hydrolysis.
This review addresses the regulatory consequences of the binding of GTP to the alpha subunits (Gα) of heterotrimeric G proteins, the reaction mechanism of GTP hydrolysis catalyzed by Gα and the means by which GTPase activating proteins (GAPs) stimulate the GTPase activity of Gα. The high energy of GTP binding is used to restrain and stabilize the conformation of the Gα switch segments, particularly switch II, to afford stable complementary to the surfaces of Gα effectors, while excluding interaction with Gβγ, the regulatory binding partner of GDP-bound Gα. Upon GTP hydrolysis, the energy of these conformational restraints is dissipated and the two switch segments, particularly switch II, become flexible and are able to adopt a conformation suitable for tight binding to Gβγ. Catalytic site pre-organization presents a significant activation energy barrier to Gα GTPase activity. The glutamine residue near the N-terminus of switch II (Glncat ) must adopt a conformation in which it orients and stabilizes the γ phosphate and the water nucleophile for an in-line attack. The transition state is probably loose with dissociative character; phosphoryl transfer may be concerted. The catalytic arginine in switch I (Argcat ), together with amide hydrogen bonds from the phosphate binding loop, stabilize charge at the β-γ bridge oxygen of the leaving group. GAPs that harbor "regulator of protein signaling" (RGS) domains, or structurally unrelated domains within G protein effectors that function as GAPs, accelerate catalysis by stabilizing the pre-transition state for Gα-catalyzed GTP hydrolysis, primarily by restraining Argcat and Glncat to their catalytic conformations. © 2016 Wiley Periodicals, Inc. Biopolymers 105: 449-462, 2016.


Mechanisms of antibiotic resistance in Burkholderia pseudomallei: implications for treatment of melioidosis.
Burkholderia pseudomallei is the etiologic agent of melioidosis. This multifaceted disease is difficult to treat, resulting in high morbidity and mortality. Treatment of B. pseudomallei infections is lengthy and necessitates an intensive phase (parenteral ceftazidime, amoxicillin-clavulanic acid or meropenem) and an eradication phase (oral trimethoprim-sulfamethoxazole). The main resistance mechanisms affecting these antibiotics include enzymatic inactivation, target deletion and efflux from the cell, and are mediated by chromosomally encoded genes. Overproduction and mutations in the class A PenA β-lactamase cause ceftazidime and amoxicillin-clavulanic acid resistance. Deletion of the penicillin binding protein 3 results in ceftazidime resistance. BpeEF-OprC efflux pump expression causes trimethoprim and trimethoprim-sulfamethoxazole resistance. Although resistance is still relatively rare, therapeutic efficacies may be compromised by resistance emergence due to increased use of antibiotics in endemic regions. Novel agents and therapeutic strategies are being tested and, in some instances, show promise as anti-B. pseudomallei infectives.


ESMO Consensus Conference on malignant lymphoma: management of 'ultra-high-risk' patients.
The European Society for Medical Oncology (ESMO) consensus conference on malignant lymphoma was held on 20 June 2015 in Lugano, Switzerland, and included a multidisciplinary panel of 25 leading experts. The aim of the conference was to develop recommendations on critical subjects difficult to consider in detail in the ESMO Clinical Practice Guidelines. The following areas were identified: (1) the elderly patient, (2) prognostic factors suitable for clinical use and (3) the 'ultra-high-risk' group. Before the conference, the expert panel was divided into three working groups; each group focused on one of these areas in order to address clinically relevant questions relating to that topic. All relevant scientific literature, as identified by the experts, was reviewed in advance. During the consensus conference, each working group developed recommendations to address each of the questions devised by their group. These recommendations were then presented to the entire multidisciplinary panel and a consensus was reached. This manuscript presents recommendations regarding the management of the following 'ultra-high-risk' situations: (1) early central nervous system relapse of diffuse large B-cell lymphoma, (2) primary refractory Hodgkin lymphoma and (3) plasmablastic lymphoma. Results, including a summary of evidence supporting each recommendation, are detailed in this manuscript. All expert panel members approved this final article.


Validation of a GC/MS method for the detection of two quinolinone-derived selective androgen receptor modulators in doping control analysis.
Selective androgen receptor modulators (SARMs) represent an emerging class of drugs likely to be abused in sport. For clinical applications, these substances provide a promising alternative to testosterone-replacement therapies and their advantages include oral bioavailability, androgen receptor specificity, tissue selectivity, and the absence of steroid-related side effects. Although not yet commercially available, since January 2008 SARMs have been included on the prohibited list issued yearly by the World Anti-Doping Agency (WADA), so control laboratories need to update their procedures to detect either the parent drugs or their metabolites. Within this context, two quinolinone SARM models were synthesized and automatically characterized to update the existing routine screening procedures. The conditions for the new target analytes are compatible with the existing laboratory protocols used for both in-competition and out-of-competition controls and can be included in them. Validation parameters according to ISO 17025 and WADA guidelines were successfully determined. For analytical determinations, spiked urine samples were hydrolyzed and extracted at pH 9.6 with 10 mL of tert-butyl methyl ether. Then, the analytes were subsequently converted into trimethylsilyl derivatives and detected by gas chromatography-mass spectrometry. The absence of interferents, together with excellent repeatability of both retention times and the relative abundances of diagnostic ions, allowed proper identification of all SARM analytes. The analytes' quantification was linear up to 500 ng/mL and precision criteria were satisfied (coefficient of variation less than 25% at 10 ng/mL). The limits of detection were 1 ng/mL for both SARMs, whereas recovery values were between 95.5 and 99.3%. The validated method can be efficiently used for urine screening of the 2-quinolinone-derived SARMs tested.


The function of multiple ejaculations in bitterling.
In some taxa, males perform multiple ejaculations, which may function in sperm competition or in maintaining a baseline density of spermatozoa in the female reproductive tract to ensure fertilization, a process that has been termed 'topping up'. We investigated the function of multiple ejaculations in two species of bitterling, the European bitterling (Rhodeus amarus) and Chinese rose bitterling (Rhodeus ocellatus). Bitterling oviposit in living freshwater mussels, with fertilization taking place within the mussel gill cavity. Thus, although fertilization is external, the mussel is analogous to the female reproductive tract in an internally fertilizing species. We measured the frequency of ejaculations and mussel inspections by individual males of two bitterling species in 28 replicated mesocosms and examined focal male responses to rival ejaculations and the presence of females in spawning condition. We used a model of ejaculatory behaviour to simulate the temporal abundance of spermatozoa in mussels. Male R. amarus exhibited high rates of ejaculation and inspection of the siphons of mussels and increased their ejaculation rate in response to the presence of females in spawning condition. Rhodeus ocellatus showed lower overall rates of ejaculation, but significantly elevated ejaculation rate in response to rival ejaculations. The ejaculatory strategy of R. amarus is one that maintains a minimum level of spermatozoa in mussels, which is elevated when the probability of oviposition increases. In contrast, R. ocellatus engages more directly in sperm competition with rivals. We discuss these results in the context of the function of multiple ejaculations and male mating tactics.


Neisseria gonorrhoeae clustering to reveal major European whole-genome-sequencing-based genogroups in association with antimicrobial resistance.
Neisseria gonorrhoeae, the bacterium responsible for the sexually transmitted disease gonorrhoea, has shown an extraordinary ability to develop antimicrobial resistance (AMR) to multiple classes of antimicrobials. With no available vaccine, managing N. gonorrhoeae infections demands effective preventive measures, antibiotic treatment and epidemiological surveillance. The latter two are progressively being supported by the generation of whole-genome sequencing (WGS) data on behalf of national and international surveillance programmes. In this context, this study aims to perform N. gonorrhoeae clustering into genogroups based on WGS data, for enhanced prospective laboratory surveillance. Particularly, it aims to identify the major circulating WGS-genogroups in Europe and to establish a relationship between these and AMR. Ultimately, it enriches public databases by contributing with WGS data from Portuguese isolates spanning 15 years of surveillance. A total of 3791 carefully inspected N. gonorrhoeae genomes from isolates collected across Europe were analysed using a gene-by-gene approach (i.e. using cgMLST). Analysis of cluster composition and stability allowed the classification of isolates into a two-step hierarchical genogroup level determined by two allelic distance thresholds revealing cluster stability. Genogroup clustering in general agreed with available N. gonorrhoeae typing methods [i.e. MLST (multilocus sequence typing), NG-MAST (N. gonorrhoeae multi-antigen sequence typing) and PubMLST core-genome groups], highlighting the predominant genogroups circulating in Europe, and revealed that the vast majority of the genogroups present a dominant AMR profile. Additionally, a non-static gene-by-gene approach combined with a more discriminatory threshold for potential epidemiological linkage enabled us to match data with previous reports on outbreaks or transmission chains. In conclusion, this genogroup assignment allows a comprehensive analysis of N. gonorrhoeae genetic diversity and the identification of the WGS-based genogroups circulating in Europe, while facilitating the assessment (and continuous monitoring) of their frequency, geographical dispersion and potential association with specific AMR signatures. This strategy may benefit public-health actions through the prioritization of genogroups to be controlled, the identification of emerging resistance carriage, and the potential facilitation of data sharing and communication.


Novel ABHD12 mutations in PHARC patients: the differential diagnosis of deaf-blindness.
This study examines ABHD12 mutation analysis in 2 PHARC patients, originally thought to be Usher syndrome. The ABHD12 gene of 2 patients, who suffered from deaf-blindness and dysfunctional central and peripheral nervous systems, were sequenced. We identified that both cases carried the same novel splice site mutation in the ABHD12 gene. However, 1 had epilepsy and the other had peripheral neuropathy. Based on haplotype analysis, the mutation is likely not a hot spot, but rather could be attributable to a common ancestor. This study shows that PHARC has phenotypic variability, even within a family, which is consistent with previous reports. Differential diagnosis of "deaf-blindness" diseases is crucial. Confirming the presence of associated symptoms is necessary for differentiating some deaf-blindness syndromes. In addition, mutation analysis is a useful tool for confirming the diagnosis.


An optimized activity-based probe for the study of caspase-6 activation.
Although significant efforts have been made to understand the mechanisms of caspase activation during apoptosis, many questions remain regarding how and when executioner caspases get activated. We describe the design and synthesis of an activity-based probe that labels caspase-3/-6/-7, allowing direct monitoring of all executioner caspases simultaneously. This probe has enhanced in vivo properties and reduced cross-reactivity compared to our previously reported probe, AB50. Using this probe, we find that caspase-6 undergoes a conformational change and can bind substrates even in the absence of cleavage of the proenzyme. We also demonstrate that caspase-6 activation does not require active caspase-3/-7, suggesting that it may autoactivate or be cleaved by other proteases. Together, our results suggest that caspase-6 activation proceeds through a unique mechanism that may be important for its diverse biological functions.


Revising or redoing an imperfect or failed nasal reconstruction.
Almost all major nasal reconstructions will require a late revision to refine aesthetics and function. The early surgical result after pedicle division will be determined by the materials, methods, priorities, planning, and surgical stages chosen by the surgeon. Imperfections in nasal contour, including recreation of the alar crease and nasolabial fold, are corrected by soft tissue debulking and secondary cartilage grafting through peripheral or direct incisions. The nostrils are enlarged by soft tissue excision and local tissue rearrangement. Occasionally, the original repair must be discarded and a second regional flap used to "redo" the reconstruction. Success is determined by careful analysis of the visual deformity, regional principles of the subunit reconstruction, and careful planning.


An in vitro study measuring marginal gaps of inlay restorations fabricated from different CAD-CAM materials after thermocycling.
Many monolithic machined materials have been introduced and provided a suitable mechanical and physical properties for inlay restorations. However, there is shortage in the studies evaluating the marginal adaptation using these materials. This study aimed to compare the effect of fabricating inlay restorations from 3 different CAD-CAM materials on marginal gaps before and after thermocycling. Sixty human premolars were randomly divided into 3 groups (n = 20) according to the material used: (e.max CAD, Ivoclar AG, Schaan, Liechtenstein), (HC, Shofu, Koyoto, Japan) and (Brilliant Crios, Coltene, Altstätten, Switzerland) (n = 20). A scanning electron microscope (SEM) (JSM- 6510 lv, JEOL, Tokyo, JAPAN) was used to for measuring the marginal gaps after cementation of inlay restorations. The magnification was adapted to 250x. Marginal gaps were revaluated with SEM after thermocycling. The temperatures of baths were 5 and 55 °C was applied for a total of 5000 cycles. All data were statistically analyzed by using ANCOVA to demonstrate if there were any statistically significant differences between the gap measures after thermocycling of the three independent (unrelated) groups. A Bonferroni adjustmen was used to perform post hoc analysis (α = 0.05). Post-intervention marginal gap was statistically significantly lower in group EX (110.8 μm) which was statistically significant compared with group SF (112.5 μm) (mean difference=-1.768, P = .007) and group BR (113 μm) (mean difference=-2.272, P = .001), however, in. comparing SF and BR groups, there was no significant difference (mean difference=-0.5, P = .770). Thermocycling affected the marginal gaps of composite based restoration and resin-modified ceramics widely. However, it had a very small effect on glass ceramics marginal adaptation. The marginal gaps of CAD-CAM inlays varied according to material used (ceramic based, combination, or resin based). Thermocycling has a minor effect on the marginal adaptation of lithium disilicate glass-ceramic inlays, where it affected the margin of resin-modified ceramic and composite based inlays greatly. Using lithium disilicate glass-ceramic might improve the clinical longevity of inlay restored teeth.


Mismatch repair gene mutations lead to lynch syndrome colorectal cancer in rhesus macaques.
Colorectal cancer accounts for a substantial number of deaths each year worldwide. Lynch Syndrome is a genetic form of colorectal cancer (CRC) caused by inherited mutations in DNA mismatch repair (MMR) genes. Although researchers have developed mouse models of Lynch Syndrome through targeted mutagenesis of MMR genes, the tumors that result differ in important ways from those in Lynch Syndrome patients. We identified 60 cases of CRC in rhesus macaques (Macaca mulatta) at our facility since 2001. The tumors occur at the ileocecal junction, cecum and proximal colon and display clinicopathologic features similar to human Lynch Syndrome. We conducted immunohistochemical analysis of CRC tumors from several rhesus macaques, finding they frequently lack expression of MLH1 and PMS2 proteins, both critical MMR proteins involved in Lynch Syndrome. We also found that most macaque cases we tested exhibit microsatellite instability, a defining feature of Lynch Syndrome. Whole genome sequencing of rhesus macaque CRC cases identified mutations in MLH1 and/or MSH6 that are predicted to disrupt protein function. We conclude that this population of rhesus macaques constitutes a spontaneous model of Lynch Syndrome, matching the human disease in several significant characteristics, including genetic risk factors that parallel human Lynch Syndrome.


EVOTECH endoscope cleaner and reprocessor (ECR) simulated-use and clinical-use evaluation of cleaning efficacy.
The objective of this study was to perform simulated-use testing as well as a clinical study to assess the efficacy of the EVOTECH Endoscope Cleaner and Reprocessor (ECR) cleaning for flexible colonoscopes, duodenoscopes, gastroscopes and bronchoscopes. The main aim was to determine if the cleaning achieved using the ECR was at least equivalent to that achieved using optimal manual cleaning. Simulated-use testing consisted of inoculating all scope channels and two surface sites with Artificial Test Soil (ATS) containing 108 cfu/mL of Enterococcus faecalis, Pseudomonas aeruginosa and Candida albicans. Duodenoscopes, colonoscopes, and bronchoscopes (all Olympus endoscopes) were included in the simulated use testing. Each endoscope type was tested in triplicate and all channels and two surface sites were sampled for each scope. The clinical study evaluated patient-used duodenoscopes, bronchoscopes, colonoscopes, and gastroscopes (scopes used for emergency procedures were excluded) that had only a bedside flush prior to being processed in the ECR (i.e. no manual cleaning). There were 10 to 15 endoscopes evaluated post-cleaning and to ensure the entire ECR cycle was effective, 5 endoscopes were evaluated post-cleaning and post-high level disinfection. All channels and two external surface locations were sampled to evaluate the residual organic and microbial load. Effective cleaning of endoscope surfaces and channels was deemed to have been achieved if there was < 6.4 microg/cm2 of residual protein, < 1.8 microg/cm2 of residual hemoglobin and < 4 Log10 viable bacteria/cm2. Published data indicate that routine manual cleaning can achieve these endpoints so the ECR cleaning efficacy must meet or exceed these to establish that the ECR cleaning cycle could replace manual cleaning In the clinical study 75 patient-used scopes were evaluated post cleaning and 98.8% of surfaces and 99.7% of lumens met or surpassed the cleaning endpoints set for protein, hemoglobin and bioburden residuals. In the simulated-use study 100% of the Olympus colonoscopes, duodenoscopes and bronchoscopes evaluated met or surpassed the cleaning endpoints set for protein, and bioburden residuals (hemoglobin was not evaluated). The ECR cleaning cycle provides an effective automated approach that ensures surfaces and channels of flexible endoscopes are adequately cleaned after having only a bedside flush but no manual cleaning. It is crucial to note that endoscopes used for emergency procedures or where reprocessing is delayed for more than one hour MUST still be manually cleaned prior to placing them in the ECR.


Fucoxanthin prevents lipopolysaccharide-induced depressive-like behavior in mice via AMPK- NF-κB pathway.
Fucoxanthin (FX), a natural carotenoid abundant in edible brown seaweeds, has been shown the great anti-oxidant, anti-inflammatory and anti-diabetic effects in vivo and in vitro. The present study was designed to investigate the effects of FX on lipopolysaccharide (LPS)-induced behavioral defects in mice. In depressive behavior tests, the increased immobility time of forced swimming test and tail suspension test by LPS treatment in mice, which were significantly reversed by FX treatment (200 mg/kg, i.g.). In anxiety behavior tests, LPS injection was neither influence the anxiety-related parameters in marble burying test nor that in elevated plus maze test. Interestingly, anxiolytic effects were observed in single FX treated control and LPS-induced mice groups. FX treatment also reversed LPS-induced body weight loss and food intake decreases. Biochemical analysis indicated that FX inhibited LPS-induced overexpression of pro-inflammatory cytokines (IL-1β, IL-6 and TNF-α), as well as iNOS and COX-2 in the hippocampus, frontal cortex and hypothalamus, via the modulation of AMPK-NF-κB signaling pathway.


Cargo regulates clathrin-coated pit invagination via clathrin light chain phosphorylation.
Clathrin light chains (CLCs) control selective uptake of a range of G protein-coupled receptors (GPCRs), although the mechanism by which this occurs has remained elusive thus far. In particular, site-specific phosphorylation of CLCb controls the uptake of the purinergic GPCR P2Y12, but it is dispensable for the constitutive uptake of the transferrin receptor (TfR). We demonstrate that phosphorylation of CLCb is required for the maturation of clathrin-coated pits (CCPs) through the transition of flat lattices into invaginated buds. This transition is dependent on efficient clathrin exchange regulated by CLCb phosphorylation and mediated through auxilin. Strikingly, this rearrangement is required for the uptake of P2Y12 but not TfR. These findings link auxilin-mediated clathrin exchange to early stages of CCP invagination in a cargo-specific manner. This supports a model in which CCPs invaginate with variable modes of curvature depending on the cargo they incorporate.


Depression and use of a health plan-sponsored physical activity program by older adults.
The purpose of this study was to determine the associations between depression and the likelihood of enrollment in a health plan-sponsored physical activity program and pattern of program participation over 2 years; a secondary aim was to examine the association between participation dose and depression risk. There are no published studies on how depression influences participation in health plan-sponsored physical activity programs and how participation affects depression risk in older adults in nonresearch settings. This study used administrative data from a Medicare Advantage plan. Participants (n=4766) were enrolled in the plan for at least 1 year prior to participating in the plan-sponsored health club benefit (Silver Sneakers). Controls were age- and gender-matched to participants (n=9035). Members were identified as having depression based on ICD-9-CM codes. Multivariate regression and generalized estimating equations models were used. Data were collected between 1998 and 2003 and analyzed in 2008. Members who had a history of depression were as likely to participate in Silver Sneakers as nondepressed members (OR: 1.03; 95% CI=0.89, 1.20; p=0.67). The risk of lapse in Silver Sneakers attendance was 28%-55% (p<0.05) higher for depressed participants during months 15-24. For nondepressed Silver Sneakers participants, attendance of at least 2 visits/week during Year 1 was significantly associated with lower risk of depression in Year 2 (OR=0.54; 95% CI=0.37, 0.79; p=0.002); a similar but statistically nonsignificant association was observed for previously depressed participants (OR=0.51; 95% CI=0.26, 1.02; p=0.06). While depressed older adults are as likely to enroll in a health plan-sponsored physical activity as nondepressed members, they were at higher risk of attendance lapses. Greater participation in the physical activity program was associated with lower depression risk.


[Finerenone (Kerendia®), a new weapon against the chronic kidney disease of a patient with type 2 diabetes].
Finerenone, a new nonsteroidal mineralocorticoid receptor antagonist, showed a significant reduction in a primary composite renal outcome in FIDELIO-DKD and a significant reduction in a primary composite cardiovascular outcome in FIGARO-DKD in patients with type 2 diabetes (T2D) and a chronic kidney disease (CKD). In a subsequent analysis that combined these two clinical trials (FIDELITY), the reduction becomes statistically significant when compared to placebo for both outcomes, with a hazard ratio of 0.86 (95 % confidence interval 0.78-0.95; P = 0.0018) for the cardiovascular outcome and 0.77 (0.67-0.88; P = 0.0002) for the renal outcome. Furthermore, all renal events occurred less frequently with finerenone than with placebo, including the progression to end-stage CKD independently of the baseline levels of glomerular filtration rate and albuminuria and regardless of associated medications (including gliflozins). The safety profile was excellent. However, a significant increase in serum potassium level was observed. Even if it is less pronounced than the increase usually seen with spironolactone, the risk of hyperkalemia requires some caution regarding both patient selection and monitoring. Finerenone (Kerendia®) is indicated in the treatment of CKD with albuminuria in adult patients with T2D. In Belgium, it is reimbursed with conditions in combination with a renin-angiotensin blocker.


Mathematical Approach to Investigate Stress due to Control Measures to Curb COVID-19.
COVID-19 is a world pandemic that has affected and continues to affect the social lives of people. Due to its social and economic impact, different countries imposed preventive measures that are aimed at reducing the transmission of the disease. Such control measures include physical distancing, quarantine, hand-washing, travel and boarder restrictions, lockdown, and the use of hand sanitizers. Quarantine, out of the aforementioned control measures, is considered to be more stressful for people to manage. When people are stressed, their body immunity becomes weak, which leads to multiplying of coronavirus within the body. Therefore, a mathematical model consisting of six compartments, Susceptible-Exposed-Quarantine-Infectious-Hospitalized-Recovered (SEQIHR) was developed, aimed at showing the impact of stress on the transmission of COVID-19 disease. From the model formulated, the positivity, bounded region, existence, uniqueness of the solution, the model existence of free and endemic equilibrium points, and local and global stability were theoretically proved. The basic reproduction number (R 0) was derived by using the next-generation matrix method, which shows that, when R 0 < 1, the disease-free equilibrium is globally asymptotically stable whereas when R 0 > 1 the endemic equilibrium is globally asymptotically stable. Moreover, the Partial Rank Correlation Coefficient (PRCC) method was used to study the correlation between model parameters and R 0. Numerically, the SEQIHR model was solved by using the Rung-Kutta fourth-order method, while the least square method was used for parameter identifiability. Furthermore, graphical presentation revealed that when the mental health of an individual is good, the body immunity becomes strong and hence minimizes the infection. Conclusively, the control parameters have a significant impact in reducing the transmission of COVID-19.


Materials: marangoni convection in space microgravity environments.
Thermally induced surface-driven convection (thermal Marangoni convection) was investigated in the Spacelab 1 microgravity environment. The configuration studied is related to the floating-zone technique used for crystal growth.


[Programmed relaparotomy in generalized purulent peritonitis].
Efficacy of programmed relaparotomy in combined treatment of 51 patients with generalized peritonitis is analyzed. Based on evaluation of surgical data, basic clinical and laboratory markers of endotoxicosis algorithm of programmed relaparotomy has been developed. Comparative analysis of the results of treatment of 36 patients after 1-2 programmed relaparotomies (1.6+/-0.5 surgeries) and 15 patients after repeated relaparotomies (3.4+/-0.3 surgeries) demonstrated that these methods were more effective before development of significant polyorganic disturbances. Nine (17.6%) patients died.


Fetal Blood Flow and Genetic Mutations in Conotruncal Congenital Heart Disease.
In congenital heart disease, the presence of structural defects affects blood flow in the heart and circulation. However, because the fetal circulation bypasses the lungs, fetuses with cyanotic heart defects can survive in utero but need prompt intervention to survive after birth. Tetralogy of Fallot and persistent truncus arteriosus are two of the most significant conotruncal heart defects. In both defects, blood access to the lungs is restricted or non-existent, and babies with these critical conditions need intervention right after birth. While there are known genetic mutations that lead to these critical heart defects, early perturbations in blood flow can independently lead to critical heart defects. In this paper, we start by comparing the fetal circulation with the neonatal and adult circulation, and reviewing how altered fetal blood flow can be used as a diagnostic tool to plan interventions. We then look at known factors that lead to tetralogy of Fallot and persistent truncus arteriosus: namely early perturbations in blood flow and mutations within VEGF-related pathways. The interplay between physical and genetic factors means that any one alteration can cause significant disruptions during development and underscore our need to better understand the effects of both blood flow and flow-responsive genes.


Vasculitis: classification and management.
Vasculitis, which is pathologically an inflammatory or necrotic state of blood vessels, consists of a diverse group of disesases. The clinical manifestations of vasculitis vary greatly in severity. The simplified classification described in this article is geared toward recognizing the degree of severity of the vasculitic process so that appropriate management can be initiated expeditiously. Emphasis is placed on the vasculitides likely to be encountered in general practice.


Determination of vanillin in orange, grapefruit, tangerine, lemon, and lime juices using GC-olfactometry and GC-MS/MS.
The presence of vanillin in orange, tangerine, lemon, lime, and grapefruit juices has been identified and confirmed using high-resolution GC retention index values, mass spectra, and aroma quality. The impact of vanillin on the flavor score for grapefruit juice is discussed and reported to be minimal. Vanillin concentrations are determined to be in the low parts-per-million range for the various citrus juices. The calculated concentrations in the orange, tangerine, lemon, lime, and grapefruit juices are 0.20, 0.35, 0.41, 0.35, and 0.60 ppm, respectively. Pasteurization produced an average 15% increase in the concentration of vanillin in grapefruit juices. Vanillin did not correlate well with the overall flavor score despite a rather intense signal using OSME gas chromatography-olfactometry software.


Dermatologic surgery--its scope and some practical aspects.
Dermatologic surgery is of increasing interest to both dermatologists and other medical practitioners. It includes procedure like biopsy, excisional surgery, laser surgery, electrosurgery, curettage surgery, nail surgery, cryosurgery, minigrafting, sclerotherapy, collagen implant, punch grafting, Moh's micrographic surgery, dermabrasion and hair transplant. This article highlights the important aspects of the various procedures and some of the more important practical points.


R-carvedilol, a potential new therapy for Alzheimer's disease.
For decades, the amyloid cascade hypothesis has been the leading hypothesis in studying Alzheimer's disease (AD) pathology and drug development. However, a growing body of evidence indicates that simply removing amyloid plaques may not significantly affect AD progression. Alternatively, it has been proposed that AD progression is driven by increased neuronal excitability. Consistent with this alternative hypothesis, recent studies showed that pharmacologically limiting ryanodine receptor 2 (RyR2) open time with the R-carvedilol enantiomer prevented and reversed neuronal hyperactivity, memory impairment, and neuron loss in AD mouse models without affecting the accumulation of ß-amyloid (Aβ). These data indicate that R-carvedilol could be a potential new therapy for AD.


Tests for presynaptic modulation of corticospinal terminals from peripheral afferents and pyramidal tract in the macaque.
The efficacy of sensory input to the spinal cord can be modulated presynaptically during voluntary movement by mechanisms that depolarize afferent terminals and reduce transmitter release. It remains unclear whether similar influences are exerted on the terminals of descending fibres in the corticospinal pathway of Old World primates and man. We investigated two signatures of presynaptic inhibition of the macaque corticospinal pathway following stimulation of the peripheral nerves of the arm (median, radial and ulnar) and the pyramidal tract: (1) increased excitability of corticospinal axon terminals as revealed by changes in antidromically evoked cortical potentials, and (2) changes in the size of the corticospinal monosynaptic field potential in the spinal cord. Conditioning stimulation of the pyramidal tract increased both the terminal excitability and monosynaptic fields with similar time courses. Excitability was maximal between 7.5 and 10 ms following stimulation and returned to baseline within 40 ms. Conditioning stimulation of peripheral nerves produced no statistically significant effect in either measure. We conclude that peripheral afferents do not exert a presynaptic influence on the corticospinal pathway, and that descending volleys may produce autogenic terminal depolarization that is correlated with enhanced transmitter release. Presynaptic inhibition of afferent terminals by descending pathways and the absence of a reciprocal influence of peripheral input on corticospinal efficacy would help to preserve the fidelity of motor commands during centrally initiated movement.


[Contemporary pharmacogenetic approaches to the treatment of age-related macular degeneration].
A study on the role of CFH, HTRA and IL-8 gene polymorphism in age-related macular degeneration (AMD) development has been conducted. At the first stage of the study genetic testing was done in 69 patients with exudative AMD and 370 random Moscow citizens without the disease. The goal of the second stage was to determine the influence of gene polymorphism on patient's response to endovitreal ranibizumab treatment. For that, visual acuity and foveal thickness were assessed before and after ranibizumab injections in 120 patients with wet AMD. All patients were genotyped for the genes of interest. The results showed that the presence of homozygous 402H polymorphism in CFH gene, as well as homozygous (-625)A mutation in HTRA1 gene, determines certain clinical presentations. Moreover, visual acuity below 0.1 and presence of 402H, (-625)A and (-251)A alleles in both copies of all three genes (CFH, HTRA and IL-8) are negative predictors of disease severity and antiangiogenic treatment response.


Chronic intermittent hypoxia upregulates genes of lipid biosynthesis in obese mice.
Obstructive sleep apnea (OSA), a condition tightly linked to obesity, leads to chronic intermittent hypoxia (CIH) during sleep. There is emerging evidence that OSA is independently associated with insulin resistance and fatty liver disease, suggesting that OSA may affect hepatic lipid metabolism. To test this hypothesis, leptin-deficient obese (ob/ob) mice were exposed to CIH during the light phase (9 AM-9 PM) for 12 wk. Liver lipid content and gene expression profile in the liver (Affymetrix 430 GeneChip with real-time PCR validation) were determined on completion of the exposure. CIH caused a 30% increase in triglyceride and phospholipid liver content (P < 0.05), whereas liver cholesterol content was unchanged. Gene expression analysis showed that CIH upregulated multiple genes controlling 1) cholesterol and fatty acid biosynthesis [malic enzyme and acetyl coenzyme A (CoA) synthetase], 2) predominantly fatty acid biosynthesis (acetyl-CoA carboxylase and stearoyl-CoA desaturases 1 and 2), and 3) triglyceride and phospholipid biosynthesis (mitochondrial glycerol-3-phosphate acyltransferase). A majority of overexpressed genes were transcriptionally regulated by sterol regulatory element-binding protein (SREBP) 1, a master regulator of lipogenesis. A 2.8-fold increase in SREBP-1 gene expression in CIH was confirmed by real-time PCR (P = 0.001). Expression of major genes of cholesterol biosynthesis, SREBP-2 and 3-hydroxy-3-methylglutaryl-CoA reductase, was unchanged. In conclusion, we have shown that CIH may exacerbate preexisting fatty liver of obesity via upregulation of the pathways of lipid biosynthesis in the liver.


Decreased neurokinin-1 (substance P) receptor binding in patients with panic disorder: positron emission tomographic study with [18F]SPA-RQ.
Positron emission tomography (PET) can localize and quantify neurokinin-1 (NK(1)) receptors in brain using the nonpeptide antagonist radioligand, [(18)F]SPA-RQ. We sought to determine if patients with panic disorder have altered density of NK(1) receptors in brain because of their history of recurrent panic attacks. We also sought to determine if a drug-induced panic attack releases substance P in brain, as measured by decreased binding of [(18)F]SPA-RQ. Positron emission tomography scans with [(18)F]SPA-RQ were performed in 14 patients with panic disorder and 14 healthy subjects. Of these two groups, 7 patients and 10 healthy subjects were scanned twice, once at baseline and once after injection of doxapram, a drug that induces panic attacks. NK(1) receptor binding in patients (n = 14) compared with that in healthy subjects (n = 14) was significantly decreased by 12% to 21% in all brain regions. Doxapram effectively produced panic attacks in 6 of 7 patients with panic disorder but only 2 of 10 healthy subjects. Doxapram caused no significant change of [(18)F]SPA-RQ binding in either patients or healthy subjects. Although induction of a panic attack has no significant effect on [(18)F]SPA-RQ binding to NK(1) receptors, patients with panic disorder have widespread reduction of NK(1) receptor binding in brain.


Electron contamination and build-up doses in conformal radiotherapy fields.
The dose in the build-up region depends upon the primary photon beam, backscattered radiation from the patient and contamination radiation from outside the patient. In this paper, a model based on measured data is proposed which allows the build-up dose for arbitrarily shaped treatment fields to be determined. The dose in the build-up region is assumed to comprise a primary photon component and a contamination component that is a function of the field size and shape. This contamination component, for modelling purposes, is subdivided into contributions that correspond to elements of 1 cm by 1 cm cross-sectional area at the plane of the isocentre. The magnitude of these components has been obtained by fitting measured data to an exponential function. The exponent was found to vary linearly with depth for energies between 4 MV and 20 MV. The coefficient decreased linearly with depth at 4, 6 and 8 MV, but exhibited a broad build-up region at 20 MV. The primary component, in the build-up region, could be approximated by a 100 - (100 - PSD) e(-mu d) function, where PSD is the primary surface dose. The values obtained during the fitting procedure were used to calculate dose in the build-up region for arbitrarily shaped fields. Good agreement was found in each case.


Arterial baroreflex sensitivity, plasma catecholamines, and pressor responsiveness in essential hypertension.
Arterial baroreflex sensitivity, plasma norepinephrine (NE) and epinephrine (E), and pressor and depressor responses were assessed in 25 patients with essential hypertension and 29 normotensive control subjects. Sensitivity of the cardiac limb of the baroreflex was determined by blood pressure and interbeat interval responses associated with the Valsalva maneuver, externally applied neck suction and pressure, and injection of phenylephrine and nitroglycerin. By all these techniques, patients with essential hypertension had significantly decreased baroreflex sensitivity, even after adjustment for age mismatching between the hypertensive and normotensive groups. Hypertensive patients also had significantly higher mean levels of plasma NE and E in both brachial arterial and antecubital venous blood (246 vs 154 pg/ml arterial NE, 286 vs 184 pg/ml venous NE, 99 vs 55 pg/ml arterial E, and 65 vs 35 pg/ml venous E) and significantly larger pressor responses to injected phenylephrine (30.9 mm Hg/100 micrograms vs 16.7 mm Hg/100 micrograms). When baroreflex-cardiac sensitivity values measured by the various techniques were averaged, there was a significant inverse relationship between sensitivity and venous NE and between sensitivity and pressor responsiveness. The results indicate that decreased baroreflex-cardiac sensitivity, increased sympathetic outflow, and pressor hyperresponsiveness tend to occur together in some patients with essential hypertension. Decreased arterial distensibility and altered central neural integration can account for these findings.


Bis{2-[bis-(2-hy-droxy-eth-yl)amino]-acetato-κO,N,O'}zinc(II).
In the crystal structure of the zinc(II) complex of bicine, [Zn(C(6)H(12)NO(4))(2)], the deprotonated amino acid O,N,O'-chelates to the metal atom through a carboxyl-ate O atom, a hy-droxy O atom and the N atom, the three atoms occupying fac positions of the distorted octa-hedron surrounding the metal atom. The metal atom lies on a center of inversion. The uncoordinated carboxyl-ate O atom is hydrogen bonded to the hy-droxy groups of adjacent mol-ecules, these two hydrogen bonds leading to the formation of a three-dimensional network.


Management of perioperative hypertensive urgencies with parenteral medications.
Hypertension is the major risk factor for cardiovascular (CV) disease such as myocardial infarction (MI) and stroke. This risk is well known to extend into the perioperative period. Although most perioperative hypertension can be managed with the patient's outpatient regimen, there are situations in which oral medications cannot be administered and parenteral medications become necessary. They include postoperative nil per os status, severe pancreatitis, and mechanical ventilation. This article reviews the management of perioperative hypertensive urgency with parenteral medications. A PubMed search was conducted by cross-referencing the terms "perioperative hypertension," "hypertensive urgency," "hypertensive emergency," "parenteral anti-hypertensive," and "medication." The search was limited to English-language articles published between 1970 and 2008. Subsequent PubMed searches were performed to clarify data from the initial search. As patients with hypertensive urgency are not at great risk for target-organ damage (TOD), continuous infusions that require intensive care unit (ICU) monitoring and intraarterial catheters seem to be unnecessary and a possible misuse of resources. When oral therapy cannot be administered, patients with hypertensive urgency can have their blood pressure (BP) reduced with hydralazine, enalaprilat, metoprolol, or labetalol. Due to the scarcity of comparative trials looking at clinically significant outcomes, the medication should be chosen based on comorbidity, efficacy, toxicity, and cost.


Changing the Script: Intergenerational Communication about Sexual and Reproductive Health in Niamey, Niger.
Most strategies to reduce adolescent pregnancies have been designed to educate adolescents directly about family planning (FP), while adolescents often cite peers and parents as their primary sources of sexual health information. Yet parents' lack of knowledge about sexual and reproductive health (SRH), low self-efficacy to initiate conversations, and adverse social norms act as barriers to open intergenerational communication. To better understand the normative environment influencing communication between parents and youth about FP/SRH in francophone West Africa, the USAID supported Breakthrough RESEARCH project conducted a multi-stage qualitative study in Niger. During Stage 1, the research team developed a screening tool (based on a literature review) to categorize research participants into those who practiced open intergenerational communication about FP/SRH, and those who did not. Stage 2 consisted of 40 in-depth interviews with young people (ages 15-24) and adults (≥25 years old), stratified by whether they practiced open intergenerational communication. Results showed restrictive social norms related to youth SRH and access to information and services. However, particularly among participants classified as open-communicators, there is a hierarchy of norms and normative beliefs, with abstinence as the most virtuous decision for youth, but approving communication about and access to SRH services in order to minimize harm. Participants rely on values such as the protection of youth, protection of family honor and promotion of well-being as means to act in counter-normative ways and communicate about FP/SRH. Implications for the field include demystifying and destigmatizing SRH topics, increasing adults' communication skills, and changing the "script" to a more life-affirming view of SRH.


Chitosan oligosaccharides inhibit epithelial cell migration through blockade of N-acetylglucosaminyltransferase V and branched GlcNAc structure.
Chitosan oligosaccharides (COS) have been shown to regulate various cellular and biological functions. The aim of this study was to investigate the antimetastatic potency of COS and the underlying mechanism. Here, we established a stably N-acetylglucosaminyltransferase V (GnT-V)-overexpressed MCF10A cell line. As expected, GnT-V overexpression greatly promoted cell migration in the transfectants by using wound healing assay. However, the induction in the cell migration was significantly suppressed by an addition of COS. Curiously, COS inhibited the protein expression of GnT-V in a dose dependent manner. Consistent with that, the reactivities with datura stramonium (DSA) and leuko-agglutinating phytohemagglutinin (L4-PHA) lectins, which specifically recognize branched N-acetylglucosamine (GlcNAc) structure, were also suppressed by COS. Taken together, these results demonstrated COS inhibited breast epithelial cell migration through down-regulation of GnT-V and its products, branched N-glycans, indicating that COS may serve as a potential novel therapeutic candidate for the treatment of metastatic breast cancer.


[Role of changes in the hormonal system of the gastrointestinal tract after vagotomy].
The level of gastrin, serotonin, insulin and cortisol in the blood was raised after different types of vagotomy. Raised hormone secretion depends on the time following operation, the severity of a pathological process, to a lesser degree, on the type of surgical intervention and nature of postvagotomic disturbance. Shifts in the hormonal apparatus are caused by operation and associated with it functional-morphological changes of the digestive organs, the preservation of preoperative changes and developing compensatory-adaptive reactions. An excess of intestinal hormone secretion in the postoperative period can be responsible for the development of pathological disorders, in particular the dumping syndrome.


Clinical pharmacology of platelet cyclooxygenase inhibition.
Nonsteroidal anti-inflammatory drugs and sulfinpyrazone compete dose-dependently with arachidonate for binding to platelet cyclooxygenase. Such a process closely follows systemic plasma drug concentrations and is reversible as a function of drug elimination. Peak inhibition and extent of its reversibility at 24 hr varies consistently with individual pharmacokinetic profile. Inhibition of platelet cyclooxygenase activity by these agents is associated with variable effects on prostaglandin (PG) synthesis in the gastric mucosa and the kidney. Aspirin acetylates platelet cyclooxygenase and permanently inhibits thromboxane (TX) A2 production in a dose-dependent fashion when single doses of 0.1 to 2.0 mg/kg are given. Acetylation of the enzyme by low-dose aspirin is cumulative on repeated dosing. The fractional dose of aspirin necessary to achieve a given level of acetylation by virtue of cumulative effects approximately equals the fractional daily platelet turnover. Serum TXB2 measurements obtained during long-term dosing with 0.11, 0.22, and 0.44 mg/kg aspirin in four healthy subjects could be fitted by a theoretical model assuming identical acetylation of platelet (irreversible) and megakaryocyte (reversible) cyclooxygenase. For a given dose within this range, both the rate at which cumulative acetylation occurs and its maximal extent largely depend upon the rate of platelet turnover. Continuous administration of low-dose aspirin (20 to 40 mg/day) has no statistically significant effect on urinary excretion of either 6-keto-PGF1 alpha or 2,3-dinor-6-keto-PGF1 alpha, i.e., indexes of renal and extrarenal PGI2 biosynthesis in vivo. Whether a selective sparing of extraplatelet cyclooxygenase activity by low-dose aspirin will result in increased antithrombotic efficacy, fewer toxic reactions, or both remains to be established in prospective clinical trials.


Effect of claudins 6 and 9 on paracellular permeability in MDCK II cells.
The neonatal proximal tubule has a lower permeability to chloride, higher resistance, and higher relative sodium-to-chloride permeability (P(Na)/P(Cl)) than the adult tubule, which may be due to maturational changes in the tight junction. Claudins are tight-junction proteins between epithelial cells that determine paracellular permeability characteristics of epithelia. We have previously described the presence of two claudin isoforms, claudins 6 and 9, in the neonatal proximal tubule and subsequent reduction of these claudins during postnatal maturation. The question is whether changes in claudin expression are related to changes in functional characteristics in the neonatal tubule. We transfected claudins 6 and 9 into Madin-Darby canine kidney II (MDCK II) cells and performed electrophysiological studies to determine the resultant changes in physiological characteristics of the cells. Expression of claudins 6 and 9 resulted in an increased transepithelial resistance, decreased chloride permeability, and decreased P(Na)/P(Cl) and P(HCO3)/P(Cl). These findings constitute the first characterization of the permeability characteristics of claudins 6 and 9 in a cell model and may explain why the neonatal proximal tubule has lower permeability to chloride and higher resistance than the adult proximal tubule.


Risk Factors for Perioperative Complications in Morbidly Obese Patients Undergoing Elective Posterior Lumbar Fusion.
Retrospective cohort study. The prevalence of obesity-related low back pain and degenerative disc disease is on the rise. Past studies have demonstrated that obesity is associated with higher perioperative complication rates, but there remains a gap in the literature regarding additional risk factors that further predispose this already high-risk patient population to poor surgical outcomes following elective posterior lumbar fusion (PLF). The aim of the study is to identify independent risk factors for poor 30-day perioperative outcomes in morbidly obese patients undergoing elective PLF. We identified 22 909 patients in the American College of Surgeons National Surgical Quality Improvement Program database who underwent elective PLF. There were 1861 morbidly obese patients. Baseline patient demographics and medical comorbidities were collected. Univariate analysis was performed to compare perioperative complication rates between non-morbidly obese and morbidly obese patients. The 5 most common complications in the morbidly obese group were then selected for multivariate regression analysis to identify independent risk factors for poor 30-day outcomes. Morbidly obese patients had a higher perioperative complication rate. The 5 most common complications were prolonged hospitalization, blood transfusion, readmission, wound complications, and reoperation. Independent risk factors for these complications were age ≥65 years, super obesity (ie, BMI > 48.6), chronic steroid use, American Society of Anesthesiology classification ≥3, poor functional status, long length of fusion ≥4 levels, and extended operative time (ie, operative time ≥318 minutes). Morbidly obese patients are at higher risk of perioperative complications following elective PLF. Modifiable risk factors for the most common complications are obesity and preoperative steroid use.


The control of axillary meristem fate in the maize ramosa pathway.
Plant axillary meristems are composed of highly organized, self-renewing stem cells that produce indeterminate branches or terminate in differentiated structures, such as the flowers. These opposite fates, dictated by both genetic and environmental factors, determine interspecific differences in the architecture of plants. The Cys(2)-His(2) zinc-finger transcription factor RAMOSA1 (RA1) regulates the fate of most axillary meristems during the early development of maize inflorescences, the tassel and the ear, and has been implicated in the evolution of grass architecture. Mutations in RA1 or any other known members of the ramosa pathway, RAMOSA2 and RAMOSA3, generate highly branched inflorescences. Here, we report a genetic screen for the enhancement of maize inflorescence branching and the discovery of a new regulator of meristem fate: the RAMOSA1 ENHANCER LOCUS2 (REL2) gene. rel2 mutants dramatically increase the formation of long branches in ears of both ra1 and ra2 mutants. REL2 encodes a transcriptional co-repressor similar to the TOPLESS protein of Arabidopsis, which is known to maintain apical-basal polarity during embryogenesis. REL2 is capable of rescuing the embryonic defects of the Arabidopsis topless-1 mutant, suggesting that REL2 also functions as a transcriptional co-repressor throughout development. We show by genetic and molecular analyses that REL2 physically interacts with RA1, indicating that the REL2/RA1 transcriptional repressor complex antagonizes the formation of indeterminate branches during maize inflorescence development. Our results reveal a novel mechanism for the control of meristem fate and the architecture of plants.


The Effects of NT-1044, a Novel AMPK Activator, on Endometrial Cancer Cell Proliferation, Apoptosis, Cell Stress and In Vivo Tumor Growth.
Anti-diabetic biguanide drugs such as metformin may have anti-tumorigenic effects by behaving as AMPK activators and mTOR inhibitors. Metformin requires organic cation transporters (OCTs) for entry into cells, and NT-1044 is an AMPK activator designed to have greater affinity for two of these transporters, OCT1 and OCT3. We sought to compare the effects of NT-1044 on cell proliferation in human endometrial cancer (EC) cell lines and on tumor growth in an endometrioid EC mouse model. Cell proliferation was assessed in two EC cell lines, ECC-1 and Ishikawa, by MTT assay after exposure to NT-1044 for 72 hours of treatment. Apoptosis was analyzed by Annexin V-FITC and cleaved caspase 3 assays. Cell cycle progression was evaluated by Cellometer. Reactive oxygen species (ROS) were measured using DCFH-DA and JC-1 assays. For the in vivo studies, we utilized the LKB1fl/flp53fl/fl mouse model of endometrioid endometrial cancer. The mice were treated with placebo or NT-1044 or metformin following tumor onset for 4 weeks. NT-1044 and metformin significantly inhibited cell proliferation in a dose-dependent manner in both EC cell lines after 72 hours of exposure (IC50 218 μM for Ishikawa; 87 μM for ECC-1 cells). Treatment with NT-1044 resulted in G1 cell cycle arrest, induced apoptosis and increased ROS production in both cell lines. NT-1044 increased phosphorylation of AMPK and decreased phosphorylation of S6, a key downstream target of the mTOR pathway. Expression of the cell cycle proteins CDK4, CDK6 and cyclin D1 decreased in a dose-dependent fashion while cellular stress protein expression was induced in both cell lines. As compared to placebo, NT-1044 and metformin inhibited endometrial tumor growth in obese and lean LKB1fl/flp53fl/fl mice. NT-1044 suppressed EC cell growth through G1 cell cycle arrest, induction of apoptosis and cellular stress, activation of AMPK and inhibition of the mTOR pathway. In addition, NT-1044 inhibited EC tumor growth in vivo under obese and lean conditions. More work is needed to determine if this novel biguanide will be beneficial in the treatment of women with EC, a disease strongly impacted by obesity and diabetes.


No Association between Daily Walking and Knee Structural Changes in People at Risk of or with Mild Knee Osteoarthritis. Prospective Data from the Multicenter Osteoarthritis Study.
We investigated the association between objectively measured daily walking and knee structural change, defined either as radiographic worsening or as cartilage loss, in people at risk of or with knee osteoarthritis (OA). Participants from the Multicenter Osteoarthritis Study (MOST) with Kellgren-Lawrence grades 0-2 and daily walking (measured with the StepWatch) at the 60-month visit were included. Participants had fixed-flexion, weight-bearing radiographs and knee magnetic resonance images (MRI) at 60 and 84 months. Radiographic worsening was read in both knees using the Osteoarthritis Research Society International grading, and MRI were read for 1 knee using the Whole-Organ MRI Score semiquantitative scoring. OR and 95% CI were calculated comparing those in the middle tertile against the lowest and highest tertiles of daily walking using logistic regression models and generalized estimating equations. Data on walking with moderate to vigorous intensity (min with > 100 steps/min/day) were associated to structural change using multivariate and logistic regression models. The 1179 study participants (59% women) were 67.0 years old (± 7.6), with a mean (± SD) body mass index of 29.8 kg/m(2) (± 5.3) who walked 6981 (± 2630) steps/day. After adjusting for confounders, we found no significant associations between daily walking and radiographic worsening or cartilage loss. More time spent walking at a moderate to vigorous intensity was not associated with either radiographic worsening or cartilage loss. Results from the MOST study indicated no association between daily walking and structural changes over 2 years in the knees of people at risk of or with mild knee OA.


A new method of synthesis of fluorescently labelled oligonucleotides and their application in DNA sequencing.
A new approach to the chemical synthesis of oligodeoxynucleotides bearing reporter functional groups at base residues of 3'-end nucleosides is reported. Applications of the 3'-end fluorescently labelled primers for automated DNA sequencing are shown.


[Development of Human Vital Signs and Body Posture Monitoring and Positioning Alarm Systems].
In view of the high incidence of malignant diseases such as malignant arrhythmias in the elderly population, accidental injuries such as falls, and the problem of no witnesses when danger occurs, the study developed a human vital signs and body posture monitoring and positioning alarm system. Through the collection and analysis of electrocardiogram (ECG), respiration (RESP) and acceleration (ACC) signals, the system monitors human vital signs and body posture in real time, automatically judges critical states such as malignant arrhythmias and accidental falls on the local device side, and then issues alarm information, opens the positioning function, and uploads physiological information and patient location information through 4G communication. Experiments have shown that the system can accurately determine the occurrence of ventricular fibrillation and falls, and issue position and alarm information.


Relationship of Microbial Profile With Airway Immune Response in Eosinophilic or Neutrophilic Inflammation of Asthmatics.
Different characteristics of airway microbiome in asthmatics may lead to differential immune responses, which in turn cause eosinophilic or neutrophilic airway inflammation. However, the relationships among these factors have yet to be fully elucidated. Microbes in induced sputum samples were subjected to sequence analysis of 16S rRNA. Airway inflammatory phenotypes were defined as neutrophils (>60%) and eosinophils (>3%), and inflammation endotypes were defined by levels of T helper (Th) 1 (interferon-γ), Th2 (interleukin [IL]-5 and IL-13), Th-17 (IL-17), and innate Th2 (IL-25, IL-33, and thymic stromal lymphopoietin) cytokines, inflammasomes (IL-1β), epithelial activation markers (granulocyte-macrophage colony-stimulating factor and IL-8), and Inflammation (IL-6 and tumor necrosis factor-α) cytokines in sputum supernatants was assessed by enzyme-linked immunosorbent assay. The numbers of operational taxonomic units were significantly higher in the mixed (n = 21) and neutrophilic (n = 23) inflammation groups than in the paucigranulocytic inflammation group (n = 19; p < 0.05). At the species level, Granulicatella adiacens, Streptococcus parasanguinis, Streptococcus pneumoniae, Veillonella rogosae, Haemophilus parainfluenzae, and Neisseria perflava levels were significantly higher in the eosinophilic inflammation group (n = 20), whereas JYGU_s levels were significantly higher in the neutrophilic inflammation group compared to the other subtypes (p < 0.05). Additionally, IL-5 and IL-13 concentrations were correlated with the percentage of eosinophils (p < 0.05) and IL-13 levels were positively correlated with the read counts of Porphyromonas pasteri and V. rogosae (p < 0.05). IL-1β concentrations were correlated with the percentage of neutrophils (p < 0.05). had a tendency to be positively correlated with the read count of JYGU_s (p = 0.095), and was negatively correlated with that of S. pneumoniae (p < 0.05). Difference of microbial patterns in airways may induce distinctive endotypes of asthma, which is responsible for the neutrophilic or eosinophilic inflammation in asthma.


[Effect of eosin Y on Ca2+, Mg2+-ATPase of the smooth muscle sarcolemma].
Eosin Y was studied with the aim to elucidate the mechanism of its inhibitory effect on the activity of Ca(2+)-transporting ATPase of myometrium cell plasma membrane. The inhibitor was studied for its effect on the maximal rate of the ATP-hydrolase reaction catalyzed by Ca2+, Mg(2+)-ATPase, on the enzyme affinity for the substrate and a possibility of enzyme activity protection under the inhibitor effect by the main reagents of ATP-hydrolase reaction. It was established that eosin Y decreased the turnover rate of this enzyme and his affinity for ATP. Preincubation of ATPase with ATP (or ATP plus MgCl2) had no effect on the extent of enzyme inhibition by eosin Y. This result proves that eosin Y and ATP do not compete for the site of binding on the enzyme.


Choice of random rather than imprinted X inactivation in female embryonic stem cell-derived extra-embryonic cells.
In female mammals, one of two X chromosomes is epigenetically inactivated for gene dosage compensation, known as X inactivation (Xi). Inactivation occurs randomly in either the paternal or maternal X chromosome in all embryonic cell lineages, designated as random Xi. By contrast, in extra-embryonic cell lineages, which are segregated from somatic cell lineages in pre-implantation development, the paternal X chromosome is selectively inactivated, known as imprinted Xi. Although it is speculated that erasure of the imprinted mark on either the maternal or paternal X chromosome in somatic cell lineages might change the mode of Xi from imprinted to random, it is not known when this event is completed in development. Here, we tested the mode of Xi during the differentiation of female mouse embryonic stem (ES) cells derived from the inner cell mass (ICM) of blastocyst-stage embryos toward trophectoderm (TE) and primitive endoderm (PrE) lineages induced by artificial activation of transcription factor genes Cdx2 and Gata6, respectively. We found that random Xi occurs in both TE and PrE cells. Moreover, cloned embryos generated by the transfer of nuclei from the female ES cells showed random Xi in TE, suggesting the complete erasure of all X imprints for imprinted Xi in ICM-derived ES cells.


Which are you watching, an individual reactive oxygen species or total oxidative stress?
The fluorometric measurement of an individual reactive oxygen species (ROS) provides useful biological and biochemical information on ROS as important mediators for the pathological conditions of various diseases and yet requires a highly specific probe toward the target ROS. To design such a specific ROS probe, this report proposes a new strategy based on protection-deprotection chemistry between fluoresceins and their derivatives protected with benzenesulfonyl groups. The strategy has allowed developing new fluorescent probes toward extra- and intracellular hydrogen peroxide or superoxide. Herein, I outline the strategy used for the design of these ROS probes and their probe performance with high selectivity toward hydrogen peroxide and superoxide, which could not be achieved until a simple deprotection, namely, a nonredox reaction, was used as a fluorescing process.


Effects of THC on driving performance, physiological state and subjective feelings relative to alcohol.
The effects of marijuana or THC on driving has been tested in several studies, but usually not in conjunction with physiological and subjective responses and not in comparison to alcohol effects on all three types of measures. To assess the effects of two dosages of THC relative to alcohol on driving performance, physiological strain, and subjective feelings. We tested the subjective feelings and driving abilities after placebo, smoking two dosages of THC (13 mg and 17 mg), drinking (0.05% BAC) and 24 h after smoking the high dose THC cigarette, while monitoring physiological activity of the drugs by heart rate. Fourteen healthy students, all recreational marijuana users, participated in the study. Both levels of THC cigarettes significantly affected the subjects in a dose-dependent manner. The moderate dose of alcohol and the low THC dose were equally detrimental to some of the driving abilities, with some differences between the two drugs. THC primarily caused elevation in physical effort and physical discomfort during the drive while alcohol tended to affect sleepiness level. After THC administration, subjects drove significantly slower than in the control condition, while after alcohol ingestion, subjects drove significantly faster than in the control condition. No THC effects were observed after 24 h on any of the measures.


A surprising cause of reversible dilated cardiomyopathy.
This case report describes two cases of dilated cardiomyopathy due to hypocalcaemia as a result of hypoparathyroidism. Patient A suffered from dilated cardiomyopathy due to secondary hypoparathyroidism as a result of previous neck surgery. Patient B suffered from dilated cardiomyopathy with congestive heart failure due to primary hypoparathyroidism. Hypoparathyroidism can exist for years before being recognised, especially after neck surgery. Besides standard treatment of heart failure, restoration of serum calcium levels with calcium and vitamin D supplementation can lead to rapid improvement of cardiac function and should be continued lifelong. Both patients were responding very well to heart failure therapy and calcium supplementation as ejection fraction improved after restoration of plasma calcium levels. This case report emphasises that hypocalcaemia should be in the differential diagnosis of heart failure.


Measurement of Murine Neuromuscular Function Using the In Situ Preparation.
The presence and progression of a neuromuscular pathology can impact on the contractile force production of a muscle. Hence, measurements of force production can be an important tool for the evaluation of disease progression. In this chapter, we describe how to perform in situ function testing on the tibialis anterior muscle using a murine model. Performing neuromuscular in situ function testing allows force measurements to be recorded in a physiologically relevant environment.


Combining Resampling Strategies and Ensemble Machine Learning Methods to Enhance Prediction of Neonates with a Low Apgar Score After Induction of Labor in Northern Tanzania.
The goal of this study was to establish the most efficient boosting method in predicting neonatal low Apgar scores following labor induction intervention and to assess whether resampling strategies would improve the predictive performance of the selected boosting algorithms. A total of 7716 singleton births delivered from 2000 to 2015 were analyzed. Cesarean deliveries following labor induction, deliveries with abnormal presentation, and deliveries with missing Apgar score or delivery mode information were excluded. We examined the effect of resampling approaches or data preprocessing on predicting low Apgar scores, specifically the synthetic minority oversampling technique (SMOTE), borderline-SMOTE, and the random undersampling (RUS) technique. Sensitivity, specificity, precision, area under receiver operating curve (AUROC), F-score, positive predicted values (PPV), negative predicted values (NPV) and accuracy of the three (3) boosting-based ensemble methods were used to evaluate their discriminative ability. The ensemble learning models tested include adoptive boosting (AdaBoost), gradient boosting (GB) and extreme gradient boosting method (XGBoost). The prevalence of low (<7) Apgar scores was 9.5% (n = 733). The prediction models performed nearly similar in their baseline mode. Following the application of resampling techniques, borderline-SMOTE significantly improved the predictive performance of all the boosting-based ensemble methods under observation in terms of sensitivity, F1-score, AUROC and PPV. Policymakers, healthcare informaticians and neonatologists should consider implementing data preprocessing strategies when predicting a neonatal outcome with imbalanced data to enhance efficiency. The process may be more effective when borderline-SMOTE technique is deployed on the selected ensemble classifiers. However, future research may focus on testing additional resampling techniques, performing feature engineering, variable selection and optimizing further the ensemble learning hyperparameters.