Tissue engineering strategies for promoting vascularized bone regeneration.
This review focuses on current tissue engineering strategies for promoting vascularized bone regeneration. We review the role of angiogenic growth factors in promoting vascularized bone regeneration and discuss the different therapeutic strategies for controlled/sustained growth factor delivery. Next, we address the therapeutic uses of stem cells in vascularized bone regeneration. Specifically, this review addresses the concept of co-culture using osteogenic and vasculogenic stem cells, and how adipose derived stem cells compare to bone marrow derived mesenchymal stem cells in the promotion of angiogenesis. We conclude this review with a discussion of a novel approach to bone regeneration through a cartilage intermediate, and discuss why it has the potential to be more effective than traditional bone grafting methods.


Neoadjuvant therapy for early-stage breast cancer: the clinical utility of pertuzumab.
Approximately 20% of breast cancer patients harbor tumors that overexpress human epidermal growth factor receptor 2 (HER2; also known as ErbB2), a receptor tyrosine kinase that belongs to the epidermal growth factor receptor family of receptor tyrosine kinases. HER2 amplification and hyperactivation drive the growth and survival of breast cancers through the aberrant activation of proto-oncogenic signaling systems, particularly the Ras/MAP kinase and PI3K/AKT pathways. Although HER2-positive (HER2(+)) breast cancer was originally considered to be a highly aggressive form of the disease, the clinical landscape of HER2(+) breast cancers has literally been transformed by the approval of anti-HER2 agents for adjuvant and neoadjuvant settings. Indeed, pertuzumab is a novel monoclonal antibody that functions as an anti-HER2 agent by targeting the extracellular dimerization domain of the HER2 receptor; it is also the first drug to receive an accelerated approval by the US Food and Drug Administration for use in neoadjuvant settings in early-stage HER2(+) breast cancer. Here, we review the molecular and cellular factors that contribute to the pathophysiology of HER2 in breast cancer, as well as summarize the landmark preclinical and clinical findings underlying the approval and use of pertuzumab in the neoadjuvant setting. Finally, the molecular mechanisms operant in mediating resistance to anti-HER2 agents, and perhaps to pertuzumab as well, will be discussed, as will the anticipated clinical impact and future directions of pertuzumab in breast cancer patients.


Intra-articular fate of degradable poly(ethyleneglycol)-hydrogel microspheres as carriers for sustained drug delivery.
A novel degradable microsphere (MS) for intra-articular drug delivery, composed of a polyethylene glycol (PEG) core containing degradable regions made of short poly-(lactic-co-glycolic acid) (PLGA) sequences - named PEG-hydrogel MS - was injected into the cavity of sheep shoulder joint, and compared to non-degradable MS devoid of hydrolysable crosslinker in terms of location, degradation and inflammation. One week after intra-articular injection both groups of MS were localized beneath the synovial lining of the synovial fringes located at bottom of the shoulder joint, while a fraction of particles remained in synovial fluid. Histological analyses made one and 4 weeks after intra-articular injection showed cell proliferation around the non-degradable MS entrapped within the synovium. By contrast, degradable PEG-hydrogel MS were surrounded by few cells. The degradation of degradable PEG-hydrogel MS within the synovium was slow and was not fully complete after four weeks. Our findings indicate that the tissue entrapment of MS below the synovial lining was independent of the material degradability, while degradable PEG-hydrogel MS are less inflammatory than the non-degradable one. Degradable PEG-hydrogel MS offer several advantages over the non-degradable MS as carriers for a sustained drug delivery in synovial tissue according to the low intensity of inflammatory reaction triggered in synovium.


Insulin resistance, inflammation, and non-alcoholic fatty liver disease.
Non-alcoholic fatty liver disease (NAFLD), the major cause of abnormal liver function in the western world, is often associated with obesity and diabetes. In obese individuals, fat accumulation in the abdominal region affects both lipid and glucose metabolism, and a liver loaded with fat is insulin resistant. Insulin resistance (IR) is often associated with chronic low-grade inflammation, and numerous mediators released from immune cells and adipocytes contribute to development of IR. Recent results showing an important role for these mediators in NAFLD are providing us with a better understanding of this highly prevalent disease with implications for novel therapy development. This review highlights new aspects in development of liver steatosis and the relevance of various cytokines and adipocytokines in NAFLD.


Infection control implications of the laundering of ambulance staff uniforms and reusable mops.
There is a lack of published studies on laundering in ambulance services. We performed bacterial culture on soiled and unsoiled uniforms and reusable mop heads artificially contaminated with Escherichia coli, Staphylococcus aureus, and Clostridium difficile spores. Current laundering processes used for routine cleans in the ambulances appears, from our simulations, to be effective at reducing vegetative pathogenic bacteria to undetectable levels, <3.398log10 colony-forming units (S. aureus and E. coli). Reduced levels of C. difficile were still detected after laundering but the risk this poses for infection is unknown, as background levels of these spores in the environment are unknown.


Molecular evidence for novel mercury methylating microorganisms in sulfate-impacted lakes.
Methylmercury (MeHg) is a bioaccumulative neurotoxin that is produced by certain anaerobic microorganisms, but the abundance and importance of different methylating populations in the environment is not well understood. We combined mercury geochemistry, hgcA gene cloning, rRNA methods, and metagenomics to compare microbial communities associated with MeHg production in two sulfate-impacted lakes on Minnesota's Mesabi Iron Range. The two lakes represent regional endmembers among sulfate-impacted sites in terms of their dissolved sulfide concentrations and MeHg production potential. rRNA amplicon sequencing indicates that sediments and anoxic bottom waters from both lakes contained diverse communities with multiple clades of sulfate reducing Deltaproteobacteria and Clostridia. In hgcA gene clone libraries, however, hgcA sequences were from taxa associated with methanogenesis and iron reduction in addition to sulfate reduction, and the most abundant clones were from unknown groups. We therefore applied metagenomics to identify the unknown populations in the lakes with the capability to methylate mercury, and reconstructed 27 genomic bins with hgcA. Some of the most abundant potential methylating populations were from phyla that are not typically associated with MeHg production, including a relative of the Aminicenantes (formerly candidate phylum OP8) and members of the Kiritimatiellaeota (PVC superphylum) and Spirochaetes that, together, were more than 50% of the potential methylators in some samples. These populations do not have genes for sulfate reduction, and likely degrade organic compounds by fermentation or other anaerobic processes. Our results indicate that previously unrecognized populations with hgcAB are abundant and may be important for MeHg production in some freshwater ecosystems.


Efficient inhibition of human AP endonuclease 1 (APE1) via substrate masking by abasic site-binding macrocyclic ligands.
Bis-naphthalene macrocycles, which bind with high affinity and selectivity to abasic sites in DNA, efficiently inhibit their cleavage by APE1 (IC50 = 55-60 nM in the kinetic assay with a model THF substrate). These results demonstrate that substrate masking by non-covalent abasic-site ligands is an efficient strategy for inhibition of APE1.


Molecular genetics of HLA.
A review of the molecular genetics of the HLA gene complex from a tissue typer's point of view.


Time trends in inpatient physician spending.
Despite falling admissions and declining lengths of stay, Medicare expenditures for inpatient physician services have continued to climb; this article seeks to understand this trend by examining the expenditures on a per admission basis. One hundred percent Medicare claims data were available from nine states for the 1985-1988 time period. Because Medicare's prospective payment system encourages hospitals to shift some services outside the inpatient setting, we examined trends in episodes of care, encompassing some time both before and after the inpatient stay itself. Trends were also examined at the individual DRG level in order to partially control for case-mix shifts and increased surgical use. Allowed charges were purged of both Medicare fee updates and geographic price variation in order to derive estimates of real spending growth. Hospital and physician claims were merged to form inpatient episodes that included seven days prior to admission as well as 30 days following discharge. Physician spending per episode increased 27 percent just over this four-year time period, but with considerable variation by DRG ranging from only 2 percent for transurethral prostatectomies (TURPs) to 56 percent for uncomplicated acute myocardial infarctions (AMIs). Changes in case severity and hospital and physician characteristics were all found to be important contributors to the increase in physician inpatient spending. Most important seemed to be the growth in the number of physicians associated with the inpatient stay (and the subsequent increase in diagnostic tests and other procedures). The findings suggest that control of technology and control of the number of physicians involved in the care of a patient are both critical to constraining the rate of increase in physician inpatient expenditures.


Intralesional methotrexate in the treatment of localized vitiligo: A pilot study.
Vitiligo is an immune-mediated skin disorder that targets epidermal melanocytes leading to the appearance of depigmented skin patches. Different treatment modalities have been reported with varied efficacy. We tried to evaluate the safety and efficacy of intralesional methotrexate in treating localized areas of vitiligo. Thirty participants with localized patches of vitiligo were recruited. They were treated with intralesional injections of methotrexate every 2 weeks for a maximum of six sessions. At the end of the study, the degree of repigmentation was categorized into: excellent improvement (>75% repigmentation), good improvement (50%-75% repigmentation), fair improvement (25%-50% repigmentation) and poor improvement (<25% repigmentation). We included 7 males (23.3%) and 23 females (76.7%). Their mean age was 33.6 ± 8.6 years. The duration of the disease ranged from 1 to 22 years. Four patients had a family history of vitiligo. At the end of the study, there was a highly statistically significant improvement (p < 0.001) after treatment regarding repigmentation. This study showed that intralesional methotrexate is a safe and effective treatment option for patients with localized vitiligo lesions. Further studies on a larger scale are needed to evaluate the long-term effects of treatment and detect the ideal dose to be injected.


Middle-aged rats orally supplemented with gel-encapsulated catechin favorably increases blood cytosolic NADPH levels.
Green tea catechins are primarily known to function as free radical scavengers and have several beneficial uses. Orally supplemented catechin (OSC) was previously shown to increase mitochondrial heme and catalase levels in rat heart blood, however, its effect in the cytosol has not been elucidated. Here, we determined the effects of OSC in the rat heart blood cytosol. We used middle-aged (40 week-old) and young (4 week-old) rats throughout the study. We isolated blood cytosol, verified its purity, and determined heme, hydrogen peroxide (H2O2) levels, catalase (CAT) activities, gp91(phox) amounts, NADP and NAD pools, sirtuin 1 (SIRT1) and glutathione reductase (GR) activities, and free fatty acids (FFA). We established that OSC is associated with decreased heme-dependent H2O2 amounts while increasing heme-independent CAT activity. Moreover, we found that OSC-related decrease in NAD(+) amounts among middle-aged rats is associated to increased NADPH levels and SIRT1 activity. In contrast, we associated OSC-related decrease in NAD(+) amounts among young rats to decreased NADPH levels and increased SIRT1 activity. This highlights a major difference between catechin-treated middle-aged and young rats. Furthermore, we observed that cytosolic FFA and GR levels were significantly increased only among OSC-treated middle-aged rats which we hypothesize are related to increased NADPH levels. This insinuates that OSC treatment allows higher catechin amounts to enter the bloodstream of middle-aged rats. We propose that this would favorably increase NADPH amounts and lead to the simultaneous decrease in NADPH-related pro-oxidant activity and increase in NADPH-related biomolecules and anti-oxidant activities.


Size-dependent properties and unusual reactivity of novel nonplanar heterocycloarenes.
The solution-phase synthesis of (hetero)cycloarenes with a well-defined size and geometric structure remains a challenging topic in organic chemistry and materials science. Herein, two novel nonplanar N,S-heterocycloarenes (PTZ1 and PTZ2) containing two/three alternate phenothiazine-co-phenanthrene units were conveniently synthesized. The smaller size heterocycloarene PTZ1 adopts a unique butterfly-shaped geometry and shows moderate supramolecular host-guest interactions with both fullerenes C60 and C70; whereas the higher homologue PTZ2 has a saddle-shaped conformation and demonstrates no obvious encapsulation with C60 or C70. Meanwhile, benefiting from the relatively ordered molecular packing, the thin film of PTZ1 behaved as a p-type semiconductor, while the more distorted PTZ2 does not display any field-effect characteristics. Particularly, upon the oxidation of heterocycloarene PTZ1 by Oxone, an unusual bis(sulfone-co-orthoquinone) product PTZ1-Oxi with an arc-shaped geometry is obtained and identified by single-crystal X-ray analysis. Our findings markedly expand the known chemistry of (hetero)cycloarenes and open a new path for their further functionalization.


Ischemic preconditioning of skeletal muscle: duration of late-phase protection.
The time course of the late phase of ischemic preconditioning (IPC) was determined in latissimus dorsi muscle (LDM) flaps using viability and function as the endpoints. LDM flaps from Sprague-Dawley rats were allocated into 6 groups. LDMs were preconditioned with 2 30-minute periods of ischemia separated by 10 minutes of reperfusion and subjected to a 4-hour ischemic insult after 24, 48, 72, and 96 hours from IPC. LDMs were evaluated for percent necrosis and muscle contractile function and compared with controls. The late phase of IPC provides significant protection against necrosis up to 72 hours. Conversely, when the end point used was muscle contractile function, the protection only lasted 48 hours. The time course of late-phase protection in skeletal muscle is 2-3 days. Late phase IPC appears to protect muscle flaps during the most critical time period following elevation.


Transforming growth factor-beta enhances connective tissue growth factor expression in L6 rat skeletal myotubes.
Transforming growth factor (TGF)-beta plays an important role in fibrosis of various organs and tissues. TGF-beta1 stimulates fibroblastic cells to form extracellular matrix (ECM), and regulates all critical events in wound healing. Connective tissue growth factor (CTGF), a TGF-beta-inducible molecule, has recently been reported to promote fibroblast proliferation, migration, adhesion and extracellular matrix formation, both in vivo and in vitro. In this study, we demonstrated that TGF-beta1 enhances CTGF mRNA and protein levels in L6 rat skeletal muscle myotubes. TGF-beta might, therefore, play a role in fibrosis of skeletal muscle by stimulating CTGF expression in the muscle tissue itself.


UV-photoexcitation and ultrafast dynamics of HCFC-132b (CF2 ClCH2 Cl).
The UV-induced photochemistry of HCFC-132b (CF2 ClCH2 Cl) was investigated by computing excited-state properties with time-dependent density functional theory (TDDFT), multiconfigurational second-order perturbation theory (CASPT2), and coupled cluster with singles, doubles, and perturbative triples (CCSD(T)). Excited states calculated with TDDFT show good agreement with CASPT2 and CCSD(T) results, correctly predicting the main excited-states properties. Simulations of ultrafast nonadiabatic dynamics in the gas phase were performed, taking into account 25 electronic states at TDDFT level starting in two different spectral windows (8.5 ± 0.25 and 10.0 ± 0.25 eV). Experimental data measured at 123.6 nm (10 eV) is in very good agreement with our simulations. The excited-state lifetimes are 106 and 191 fs for the 8.5 and 10.0 eV spectral windows, respectively. Internal conversion to the ground state occurred through several different reaction pathways with different products, where 2Cl, C-Cl bond breakage, and HCl are the main photochemical pathways in the low-excitation region, representing 95% of all processes. On the other hand, HCl, HF, and C-Cl bond breakage are the main reaction pathways in the higher excitation region, with 77% of the total yield. © 2015 Wiley Periodicals, Inc.


The effect of core-shell engineering on the energy product of magnetic nanometals.
Solution-based growth of magnetic FePt-FeCo (core-shell) nanoparticles with a controllable shell thickness has been demonstrated. The transition from spin canting to exchange coupling of FePt-FeCo core-shell nanostructures leads to a 28% increase in the coercivity (12.8 KOe) and a two-fold enhancement in the energy product (9.11 MGOe).


Correlation of alcohol craving with striatal dopamine synthesis capacity and D2/3 receptor availability: a combined [18F]DOPA and [18F]DMFP PET study in detoxified alcoholic patients.
In abstinent alcoholic patients, a low availability of dopamine D2/3 receptors in the ventral striatum and adjacent putamen was associated with a high level of craving for alcohol. Alcohol craving may also depend on presynaptic dysfunction of striatal dopamine production, which may contribute to the risk of relapse. In this study, positron emission tomography (PET) was used to compare dopamine synthesis capacity in the striatum in alcoholic patients and healthy comparison subjects. Positron emission tomography (PET) was used to map the net blood-brain clearance of the dopa decarboxylase substrate 6-[18F]fluoro-l-dopa, an index of dopamine synthesis capacity, in the striatum of 12 detoxified male alcoholic patients and 13 age-matched healthy men. The parametric maps were correlated with results of an earlier [18F]desmethoxyfallypride PET study of dopamine D2/3 receptor availability in the same 12 alcoholic patients and in 12 of the healthy volunteers. Alcohol craving was measured with the Alcohol Craving Questionnaire. Patients were followed for 6 months, and alcohol intake was recorded. The magnitude of net blood-brain clearance in the striatum did not differ significantly between detoxified alcoholic patients and the comparison subjects. However, a voxel-wise correlation analysis of net blood-brain clearance in the alcoholic patients linked low levels of dopamine synthesis capacity in the bilateral putamen with high levels of alcohol craving. After normalization of net blood-brain clearance maps to the voxel-wise estimates of dopamine D2/3 receptor availability, there was still a negative correlation with alcohol craving. Alcohol craving at the time of scanning was associated with high level of alcohol intake in the 6-month follow-up period. Simultaneous assay by PET of pre- and postsynaptic markers of dopamine neurotransmission indicated that a striatal dopamine deficit correlated with alcohol craving, which was associated with a high relapse risk.


A new constitutive formulation for characterizing the mechanical behavior of soft tissues.
We present a new constitutive formulation that combines certain desirable features of two previously used approaches (phenomenological and microstructural). Specifically, we assume that certain soft tissues can be idealized as composed of various families of noninteracting fibers and a homogeneous matrix. Both the fibers and the matrix are assumed to follow the gross deformation. Within the usual framework of pseudoelasticity, incompressibility, homogeneity, and the continuum hypothesis, a pseudostrain-energy function (W) is proposed wherein W is expressed in terms of matrix and fibrous contributions. Unlike phenomenological approaches where a W is usually chosen in an ad hoc manner, the present approach can be used to postulate reasonable forms of W based on limited structural information and multiaxial stress-strain data. Illustrative applications of the theory are discussed for visceral pleura and myocardium. Concise structurally motivated constitutive relations result, wherein load-dependent anisotropy, nonlinear material behavior, finite deformations, and incompressibility are accounted for.


An unusual case of Candida tropicalis sepsis in a patient submitted to allogeneic bone marrow transplantation.
We describe an exceptional case of Candida tropicalis sepsis in a patient submitted to allogeneic BMT; the diagnosis was made on a peripheral blood smear, when the pt was neutropenic and only mildly febrile. The combination of GM-CSF to accelerate hematological recovery and the possibility of administering large doses of a liposomal form of Amphotericin B were the contributing factors to the resolution of the infection.


Dimensional phenotypic analysis and functional categorisation of mutations reveal novel genotype-phenotype associations in Rett syndrome.
We aimed to improve the understanding of genotype-phenotype correlations in Rett syndrome (RS) by adopting a novel approach to categorising phenotypic dimensions - separating typicality of presentation, outcome severity and age of onset - and by classifying MECP2 mutations strictly by predicted functional attributes. MECP2 mutation screening results were available on 190 patients with a clinical diagnosis of RS (140 cases with classic RS, 50 with atypical RS). 135 cases had identified mutations. Of the 140 patients, 116 with classic RS (82.9%) had an identified mutation compared with 19 of 50 patients (38%) with an atypical presentation. Cases with early onset of regression and seizures, and those with clinical features that might indicate alternative aetiologies, were less likely to have mutations. Individuals with late truncating mutations had a less typical presentation than cases with missense and early truncating mutations, presumably reflecting greater residual function of MECP2 protein. Individuals with early truncating mutations had a more severe outcome than cases with missense and late truncating mutations. These findings held when restricting the analysis to cases over 15 years of age and classic cases only. Previous findings of variation in severity among the common mutations were confirmed. The approach to phenotypic and genotypic classification adopted here allowed us to identify genotype-phenotype associations in RS that may aid our understanding of pathogenesis and also contribute to clinical knowledge on the impact of different types of mutations.


Relapsing polychondritis.
Relapsing polychondritis is an uncommon disease consisting of inflammation of the eyes, inner ears, cardiovascular system, and cartilaginous portions of the joints, respiratory tract, and external ear. Cutaneous manifestations are the presenting feature in more than 50% of patients. These usually consist of erythema, swelling, and pain, reflecting involvement of the underlying cartilage. Direct involvement of the skin may occur as vasculitis, lesions resembling erythema nodosum, or nonspecific eruptions. The presence of circulating antibodies to type II collagen--more against native than denatured collagen--and to human fetal cartilage, and the presence of circulating immune complexes suggest a primary role for antibody in the pathogenesis of relapsing polychondritis. There is no ideal treatment, but systemic corticosteroids and dapsone seem most effective.


[Factors associated with jejunostomy complications].
There are few studies in the literature that analyze jejunostomy complications and their associated factors. To describe the rate of complications and analyze the factors associated with their development in a tertiary reference center in Mexico. A retrospective study was carried out on patients that underwent Witzel jejunostomy within the time frame of January 2002 to December 2011. Patient demographic, clinical, and laboratory data were collected at the time of hospital admission and during follow-up. The factors associated with the development of complications were analyzed using the chi square test for categorical variables and the Student's t test for the continuous variables. Statistical significance was considered with a p<.05. One hundred and twelve patients (57 men) with a mean age of 54.2 years were included in the study. Forty-eight patients (42.9%) presented with postoperative complications. The most frequent surgical complications were severe sepsis (11.6%) and septic shock (7.14%) and the most frequent medical complications were pleural effusion (7.14%) and aspiration pneumonia (6.25%). The factors that were significantly associated with the development of complications were obesity, lymphocytopenia, and hypoalbuminemia. Jejunostomy is associated with an elevated medical, as well as surgical, complication frequency. Even though it has been established as a better alternative to parenteral nutrition, its associated morbidity should be taken into consideration before establishing its routine use as a means of nutritional support, especially in patients with factors associated with the development of complications.


Alterations in regional homogeneity of resting-state brain activity in patients with major depressive disorder screening positive on the 32-item hypomania checklist (HCL-32).
Bipolar disorder (BD) is difficult to diagnose in the early stages of the illness, with the most frequent misdiagnosis being major depressive disorder (MDD). We aimed to use a regional homogeneity (ReHo) approach with resting-state functional magnetic resonance imaging (rs-fMRI) to investigate the features of spontaneous brain activity in MDD patients screening positive on the 32-item Hypomania Checklist (HCL-32). Nineteen MDD patients screening positive (HCL-32(+); 9 males; 24.9±5.7 years) and 18 patients screening negative (HCL-32(-); 9 males; 27.1±6.7 years), together with 24 healthy controls (HC; 11 males; 26.4±3.9 years) were studied. ReHo maps were compared and an receiver operating characteristic (ROC) analysis was conducted to confirm the utility of the identified ReHo differences in classifying the patients. The MDD versus HC showed different ReHo in many brain areas, especially in the frontal and parietal cortex. The HCL-32(+) versus HCL-32(-) showed significant increase of ReHo in the right medial superior frontal cortex, left inferior parietal cortex and middle/inferior temporal cortex, and decrease of ReHo in the left postcentral cortex and cerebellum. ROC analysis showed good sensitivity and specificity for distinguishing these two subgroups of MDD. Recruited patients were all on antidepressants and standard mania rating scales were not performed to assess their hypomanic symptoms. The rs-fMRI measurement of ReHo in distributed brain regions may be putative biomarkers which could differentiate subthreshold BD from MDD.


[Pulmonary eosinophilic infiltration with pneumothorax during trimipramine treatment].
Three weeks after starting treatment with the antidepressant trimipramine a 55-year-old man developed increasing cough, fatigue and lack of appetite. The blood count revealed marked eosinophilia (15%) and the chest X-ray showed bilateral pulmonary infiltrates. Later a left spontaneous pneumothorax occurred. When trimipramine was discontinued (as the suspected cause of the described changes) the symptoms quickly subsided and the infiltrate as well as the eosinophilia quickly disappeared.


A genetic linkage map of the human genome.
We report the construction of a linkage map of the human genome, based on the pattern of inheritance of 403 polymorphic loci, including 393 RFLPs, in a panel of DNAs from 21 three-generation families. By a combination of mathematical linkage analysis and physical localization of selected clones, it was possible to arrange these loci into linkage groups representing 23 human chromosomes. We estimate that the linkage map is detectably linked to at least 95% of the DNA in the human genome.


Determination of hydrogen peroxide generated by reduced nicotinamide adenine dinucleotide oxidase.
Hydrogen peroxide can be conveniently determined using horseradish peroxidase (HRP) and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid). However, interference occurs among assay components in the presence of reduced nicotinamide adenine dinucleotide (NADH) that is also a substrate of NADH oxidase. So, depletion of NADH is required before using the HRP method. Here, we report simple and rapid procedures to accurately determine hydrogen peroxide generated by NADH oxidase. All procedures developed were based on the extreme acid lability of NADH and the stability of hydrogen peroxide, because NADH was decomposed at pH 2.0 or 3.0 for 10 min, while hydrogen peroxide was stable at pH 2.0 or 3.0 for at least 60 min. Acidification and neutralization were carried out by adjusting sample containing NADH up to 30 microM to pH 2.0 for 10 min before neutralizing it back to pH 7.0. Then, hydrogen peroxide in the sample was measured using the HRP method and its determination limit was found to be about 0.3 microM. Alternatively, hydrogen peroxide in samples containing NADH up to 100 microM could be quantitated using a modified HRP method that required an acidification step only, which was found to have a determination limit of about 3 microM hydrogen peroxide in original samples.


Differentiation of cardiac and noncardiac dyspnea using bioelectrical impedance vector analysis (BIVA).
There is no gold standard for the differential diagnosis of acute dyspnea despite the usefulness of N-terminal pro-B-type natriuretic peptide (NT-proBNP) and lung ultrasound. No study has evaluated the contribution of bioelectrical impedance vector analysis (BIVA) in discriminating between cardiac and noncardiac dyspnea. We sought to determine whether a relationship exists between ultrasound detection of lung congestion, NT-proBNP, and BIVA in patients with acute dyspnea. Eligible patients were between 50 and 95 years, with an estimated glomerular filtration rate of ≥30 mL min(-1) 1.73 m(-2), who presented to an emergency department with dyspnea. Dyspnea was classified by reviewers blinded to BIVA as cardiac or noncardiac based on physical examination, electrocardiogram, chest X-ray, NT-proBNP, and B-lines of lung congestion on ultrasound. Overall, 315 patients were enrolled (median age 77 years, 48% male). An adjudicated diagnosis of cardiac dyspnea was established in 169 (54%). Using BIVA, vector positions below -1 SD of the Z-score of reactance were associated with peripheral congestion (χ(2) = 115; P < .001). BIVA measures were reasonably accurate in discriminating cardiac and noncardiac dyspnea (69% sensitivity, 79% specificity, 80% area under the receiver operating characteristic curve). In patients presenting with acute dyspnea, the combination of BIVA and lung ultrasound may provide a rapid noninvasive method to determine the cause of dyspnea.


Can financial incentives encourage women to participate in a cervical cancer screening programme? Evidence from a randomized controlled trial analysis.
The incidence of cervical cancer can be reduced by introducing effective screening programmes to detect the earliest signs of cancer. However, the literature reveals that, despite the availability of some level of infrastructural facilities for screening in low- and middle-income countries, very few women are screened in these countries due to their low acceptance of and participation in screening. Therefore, increasing the participation of women in cancer screening programmes is one of the major challenges for policy-makers. With this background, we attempted in this study to evaluate the effectiveness of financial incentives as an intervention to encourage women to accept and participate in a cervical cancer screening programme. Within a framework of randomized controlled trial (RCT) analysis, an experiment was conducted to evaluate the effectiveness of a financial intervention. The experiment was conducted in two districts of Assam, India. In total, 412 women participated in the experiment. Participants were divided between control and treatment groups on a random basis. Logistic regression was used to quantify the effectiveness of the financial intervention. The likelihood of participation in a cancer screening programme was 20.5 times greater for those respondents who received financial assistance to attend the cancer screening programme. Financial incentive was found to be an effective intervention to encourage women to participate in a cervical cancer screening programme. The study also generated evidence that, along with financial incentives, there are other factors or determinants that also play a crucial role in a woman's decision-making process. Therefore, these factors should also be considered while formulating any policies for managing and controlling cervical cancer incidence. By introducing a financial incentive as an intervention, the problem of low acceptance and participation of women in cervical cancer screening programmes can be overcome to a significant level.


The chick chorioallantoic membrane as test system for biocompatible materials.
Biologic and non-biologic materials clinically used as hemostyptica, as vascular prostheses, or as temporary skin substitutes were implanted to the chick chorioallantoic membrane (CAM) during days 9-14 of incubation. The histological study revealed an intact CAM after application of cellulose gauze (Tabotamp). The fibrin tissue adhesive (Tissucol), the collagen sponge (Tachotop), or the gelatin sponge (Gelfoam) induced different amounts of connective tissue. Some fibroblasts were about to grow into the fibrin adhesive. Fibroblasts and capillaries filled the interstices of the collagen sponge poorly, and in the case of the gelatin sponge markedly. Neutrophils and giant cells of the foreign body type occurred more often in the gelatin sponge than in the collagen sponge. With regard to the non-biologic materials, expanded polytetrafluorethylene induced squamous metaplasia of the chorionic epithelium. Dacron caused ulceration with the onset of connective tissue ingrowth, based on a reaction of giant cells of the foreign body type. Polyurethane foam (SYSpur-derm) showed bleeding into the implant due to spikes of the material. Eosinophilic granulocytes were absent in all materials studied. The results are related to different clinical findings. The correlation allows consideration of the CAM as an in vivo model in screening materials for their biocompatibility and their connective tissue reaction.


Entomophagy: Nutritional, ecological, safety and legislation aspects.
Globally, there is a need to seek alternative sources of protein in addition to meat. This has led to considerable interest in edible insects. Such insects form part of cultures and diets in many Asian and African countries, and are an excellent source of essential nutrients, minerals, vitamins and proteins. Furthermore, they have been reported to be sustainable. The ecological importance of insects is related to their short life cycles when reared and farmed. This makes them ideal in mitigating greenhouse gas emissions, cutting land uses and polluted water, and reducing environmental contamination. However, the use of edible insects as food in Europe is minimal. To ensure safety of insects when eaten as food, considerations should be made on: microbiological contamination; toxicological hazards, e.g. chemical hazards and antinutrients; allergenicity issues that are related to different exposures, including injection, ingestion, inhalation and skin contact. In this review, we summarize the nutritional and sustainable values of edible insects, look at safety and legislative measures and we finally discuss future issues.


"Age is just a number"; frailty as a marker of peri-operative risk in head and neck surgery: Systematic review and meta-analysis.
Frailty refers to a patient's reduced capacity to withstand stressors due to a reduction in physiologic reserves. We assessed the impact of frailty on outcomes following head and neck surgery. We performed a systematic review in accordance with the PRISMA guidelines. Meta-analysis was performed using the Mantel-Haenszel method. Fourteen studies incorporating 182 059 patients were included in qualitative synthesis with 15 953 (8.8%) of patients deemed as frail. Meta-analysis incorporating nine studies demonstrated that frailty is associated with an increased 30 day postoperative morbidity (OR 2.74; 95% CI 1.98-3.80; p < 0.01) and meta-analysis with six studies suggested increased 30-day mortality (OR 2.94; 95% CI 2.62-3.31; p < 0.01). Preliminary meta-analyses between two and five studies suggested that frail patients had reduced overall survival and were more likely to be discharged to a nonhome location or readmitted within 30 days. Frailty appears to be associated with poor short-term outcomes following head and neck surgery and may improve understanding of an individual patient's peri-operative risk.


Sexually dimorphic effects of maternal asthma during pregnancy on placental glucocorticoid metabolism and fetal growth.
Human pregnancy is associated with sexually dimorphic differences in mortality and morbidity of the fetus with the male fetus experiencing the poorest outcome following complications such as pre-eclampsia, pre-term delivery and infection. The physiological mechanisms that confer these differences have not been well characterised in the human. Work conducted on the effect of maternal asthma during pregnancy, combining data collected from the mother, placenta and fetus has found some significant sex-related mechanistic differences associated with fetal growth in both normal pregnancies and pregnancies complicated by asthma. Specifically, sexually dimorphic differences have been found in placental glucocorticoid metabolism in male and female fetuses of normal pregnancies. In response to the presence of maternal asthma, only the female fetus alters placental glucocorticoid metabolism resulting in decreased growth. The male fetus does not alter placental function or growth in response to maternal asthma. As a result of the alterations in glucocorticoid metabolism in the female, downstream changes occur in pathways regulated by glucocorticoids. These data suggest that the female fetus adjusts placental function and reduces growth to compensate for maternal disease. However, the male fetus continues to grow in response to maternal asthma with no changes in placental function. This response by the male fetus may partially contribute to the increased risk of morbidity and mortality in this sex.


Case report: Meningothelial hamartoma of the scalp in a 9-year-old child.
Meningothelial hamartomas represent a collection of meningothelial elements in an ectopic location. Lesions are histologically characterized as a proliferation of connective tissue elements and small- and medium-sized vessels admixed with meningothelial elements. Lesions are most often located on the scalp, present at birth, and do not extend past the subcutis. We discuss the case of a 9-year-old African American girl presenting with one such lesion on the parietal aspect of the scalp.


Early-onset versus typical childhood absence epilepsy; clinical and electrographic characteristics.
Childhood absence epilepsy (CAE) is characterized by absence attacks with stereotyped electrographic discharges. Epidemiologic data concerning early-onset CAE is scarce. We tried to specifically analyze the early-onset CAE and compare it with typical CAE. In this retrospective study, all patients with a clinical diagnosis of CAE were recruited in the outpatient epilepsy clinic at Shiraz University of Medical Sciences from 2008 to 2011. We considered the age of onset at four years or earlier as early-onset and above four years as typical CAE. Age, gender, age at seizure onset, seizure type(s), epilepsy risk factors, and EEG findings of all patients were registered routinely. Statistical analyses were performed using Chi square and Fisher's Exact tests. Forty-six patients were diagnosed as having CAE. Sixteen patients (35%) had early-onset and 30 (65%) patients had typical CAE. Sex ratio (female to male) in early-onset was 7:9 and in typical CAE was 19:11 (P=0.2). Generalized tonic-clonic and myoclonic seizures were reported in both early-onset and typical CAE. The differences were not statistically significant. Epilepsy risk factors were similarly reported in both conditions. The EEG findings were similar in both groups. We did not observe any significant differences between early-onset and typical childhood absence epilepsies with respect to the demographic, clinical and electroencephalographic characteristics.


Fat Grafting for Facial Rejuvenation with Nanofat Grafts.
Reversing structural changes in aging skin gained potential after the therapeutic use of adipose-derived stem cells was described. Nanofat is a highly concentrated solution of progenitor cells without viable adipocytes. Nanofat grafting creates striking skin quality improvement. The availability of adipose-tissue combined with straightforward mechanical protocol to process fat brings regenerative and antiaging medicine into real-life clinical practice. Association with other cofactors (hyaluronic acid, botulin toxin, and vitamin C) and therapies (microneedling and drug delivery) provides better outcomes. This article describes the techniques and the authors' experiences in nanofat grafting, and its potential new applications in regenerative medicine.


Acute hemorrhagic colitis associated with oral administration of oseltamivir for the treatment of influenza A.
The importance of oseltamivir as a strategy for the treatment of influenza has been recognized; however, the risk of acute hemorrhagic colitis should be considered, although it is rare. This report describes a case of acute hemorrhagic colitis in a 61-year-old Japanese man after the oral administration of oseltamivir for influenza A. A definitive diagnosis of influenza A was made using a rapid diagnosis kit. Abdominal pain, diarrhea, and hematochezia occurred the day after the first administration of oseltamivir (The patient had taken two capsules of oseltamivir). Endoscopic examination revealed extensive acute hemorrhagic colitis. Histopathological examination of the colonic mucosa revealed small atrophic glands with partial deciduation of the epithelium and a decrease in the number of goblet cells. Hemorrhage and mild infiltration of inflammatory cells were observed as well. These changes were suggestive of an ischemic change. A culture test showed normal flora.


Dendritic cells in contact sensitivity.
Bone-marrow-derived DC, passing through the skin or residing there as LC, acquire antigen following epicutaneous exposure to contact sensitizer. They move as veiled cells in the afferent lymphatics and migrate to draining lymph nodes, where they become interdigitating cells of the paracortex. Here they initiate T-cell responses; the cytotoxic T cells and antibody formation which develop may be able to target on DC as well as other antigen-bearing cells, so producing feed-back mechanisms to switch off immune responses. Additional features include a systemic effect which leads to movement of DC without antigen into lymph nodes. What are the signals leading to this movement and what is its significance? There is evidence for synergy between directly haptenated DC and DC not directly acquiring antigen. How does this occur and how important is this effect in ensuring the potency of DC in presenting contact sensitizer to T cells? What is the importance of antigen processing by LC? Finally, dendriform cells which may be of T-cell origin are also present in the skin. What is their role in modulating the development of contact sensitivity?


A prospective study on the use of a non-adhesive gelling foam dressing on exuding leg ulcers.
This non-comparative phase II study aimed to evaluate the safety and performance of a non-adhesive gelling foam dressing (GFD-N) in leg ulcer management. Forty-six subjects with moderately to heavily exuding leg ulcers were treated with a regimen including GFD-N. Dressings were changed at least every seven days for four weeks or until healing. Mean GFD-N wear time was 3.2 days per subject. Mean wound area decreased from 10.1 cm2 at baseline to 5.1 cm2 at four weeks (p<0.001) and healed in five subjects (11%). The surrounding skin improved or remained stable in all but one subject. When compared with pre-study dressings, ulcer pain decreased for GFD-N, both with the dressing in place (p<0.001) and on dressing removal (p<0.001). Of final investigator ratings for 45 subjects, most were 'excellent' for ease of application (89%), ease of removal (96%), conformability (67%) and overall performance (58%). Five subjects experienced adverse events; none were serious or dressing-related. This small study demonstrates that GFD-N was safe, effective and convenient for wound healing, exudate management, pain/comfort and ease of use.


Online interactive tutorials for creating graphs with excel 2007 or 2010.
Graphic display of clinical data is a useful tool for the behavior-analytic clinician. However, graphs can sometimes be difficult to create. We describe how to access and use an online interactive tutorial that teaches the user to create a variety of graphs often used by behavior analysts. Three tutorials are provided that cover the basics of Microsoft Excel 2007 or 2010, creating graphs for clinical purposes, and creating graphs for research purposes. The uses for this interactive tutorial and other similar programs are discussed.


High WSS or low WSS? Complex interactions of hemodynamics with intracranial aneurysm initiation, growth, and rupture: toward a unifying hypothesis.
Increasing detection of unruptured intracranial aneurysms, catastrophic outcomes from subarachnoid hemorrhage, and risks and cost of treatment necessitate defining objective predictive parameters of aneurysm rupture risk. Image-based computational fluid dynamics models have suggested associations between hemodynamics and intracranial aneurysm rupture, albeit with conflicting findings regarding wall shear stress. We propose that the "high-versus-low wall shear stress" controversy is a manifestation of the complexity of aneurysm pathophysiology, and both high and low wall shear stress can drive intracranial aneurysm growth and rupture. Low wall shear stress and high oscillatory shear index trigger an inflammatory-cell-mediated pathway, which could be associated with the growth and rupture of large, atherosclerotic aneurysm phenotypes, while high wall shear stress combined with a positive wall shear stress gradient trigger a mural-cell-mediated pathway, which could be associated with the growth and rupture of small or secondary bleb aneurysm phenotypes. This hypothesis correlates disparate intracranial aneurysm pathophysiology with the results of computational fluid dynamics in search of more reliable risk predictors.


Flexor digitorum profundus entrapment after closed treatment of a displaced Colles' fracture.
Osseous entrapment of the flexor digitorum profundus tendons to the long and ring fingers occurred at the fracture site after closed reduction of a comminuted, displaced Colles' fracture. The prominent tenodesis effect was relieved by tenolysis 6 months after the injury.


Stereotactic administration of intratumoral chronic chemotherapy of recurrent malignant gliomas.
There is a continuous search for more efficient treatment of malignant cerebral gliomas. The work of Penn and Croin demonstrates clearly that survival of rats having experimental gliomas is significantly increased by intratumoral chemotherapy. It has been shown that chronic depth electrodes for the investigation of epileptic patients is a reliable and safe method. Using this proven technology originally developed by Talairach, Szikla and co-workers we have developed catheters for intratumoral chemotherapy. Three patients have been treated. It is premature to pretend that this is a treatment which will lengthen the patient's life. Nevertheless, the feasibility of the technique, its precision and safety are assured. Following this course of treatment, we have seen no deteriorations, no side effects, no bone marrow depressions, and no sign of toxicity. Chemical analysis has shown that cisplatin was delivered to the tumor.


The immunomodulation of nicotinic acetylcholine receptor subunits in Zhikong scallop Chlamys farreri.
Nicotinic acetylcholine receptor (nAChR), the best-studied ionotropic neuron receptor protein, is a key player in neuronal communication, and it has been reported to play an important role in immunomodulation of vertebrates. Although nAChRs have also been identified in most invertebrates, the knowledge about their immunomodulation is still limited. In the present study, two scallop nAChR genes were identified from Chlamys farreri (designed as CfnAChR1 and CfnAChR2), which encoded 384 and 443 amino acids, respectively. The conserved disulfide-linked cystines, ion selectivity residues and the hydrophobic gating residues (L251, V255 and V259) were identified in CfnAChR1 and CfnAChR2. The immunoreactivities of CfnAChR1 and CfnAChR2 were observed in all the tested scallop tissues, including adductor muscle, mantle, gill, hepatopancreas, kidney and gonad. After LPS (0.5 mg mL(-1)) stimulation, the expression of CfnAChR1 mRNA in haemocytes increased significantly by 9.83-fold (P < 0.05) and 12.93-fold (P < 0.05) at 3 h and 24 h, respectively. While the expression level of CfnAChR2 mRNA increased 43.94% at 12 h after LPS stimulation (P < 0.05). After TNF-α (50 ng mL(-1)) stimulation, the expression levels of CfnAChR1 and CfnAChR2 both increased significantly at 1 h, which were 21.33-fold (P < 0.05) and 2.44-fold (P < 0.05) of that in the PBS group, respectively. The results collectively indicated that the cholinergic nervous system in scallops could be activated by immune stimulations through CfnAChR1 and CfnAChR2, which function as the links between the cholinergic nervous system and immune system.


Effects of exercise training on excitation-contraction coupling, calcium dynamics and protein expression in the heart of the Neotropical fish Brycon amazonicus.
Matrinxã (Brycon amazonicus) is a great swimming performance teleost fish from the Amazon basin. However, the possible cardiac adaptations of this ability are still unknown. Therefore, the aim of the present work was to investigate the effects of prolonged exercise (EX group - 60days under 0.4BL·s-1) on ventricular contractility by (i) in-vitro analysis of contractility comparing the relative roles of sodium/calcium exchanger (NCX) and sarcoplasmic reticulum (SR) in the excitation-contraction (E-C) coupling and (ii) molecular analysis of NCX, sarcoplasmic reticulum Ca2+ ATPase (SERCA2) and phospholamban (PLB) expression and quantification. The exercise training significantly improved twitch tension, cardiac pumping capacity and the contraction rate when compared to controls (CT). Inhibition of the NCX function, replacing Na+ by Li+ in the physiological solutions, diminished cardiac contractility in the EX group, reduced all analyzed parameters under both high and low stimulation frequencies. The SR blockage, using 10μM ryanodine, caused ~50% tension reduction in CT at most analyzed frequencies while in EX, reductions (34-54%) were only found at higher frequencies. SR inhibition also decreased contraction and relaxation rates in both groups. Additionally, higher post-rest contraction values were recorded for EX, indicating an increase in SR Ca2+ loading. Higher NCX and PLB expression rates and lower SERCA2 rates were found in EX. Our data indicate that matrinxã presents a modulation in E-C coupling after exercise-training, enhancing the SR function under higher frequencies. This was the first study to functionally analyze the effects of swimming-induced exercise on fish cardiac E-C coupling.


Interaction of cholinergic ligands with the purified acetylcholine receptor protein. I. Equilibrium binding studies.
We have studied the binding equilibria of two fluorescent ligands and several nonfluorescent cholinergic ligands with the purified acetylcholine receptor from Electrophorus electricus. The assay was based on the specifically cholinergic and reversible quenching of fluorescence observed upon complex formation between the receptor protein and N-7-(4-nitrobenzo-2-oxa-1,3-diazole)-5-aminopentanoic acid beta-(N-trimethylammonium) ethyl ester. This way, a large body of accurate, true equilibrium data was obtained. We find 1) all ligands studied compete for the same number of binding sites at the receptor; 2) agonists compete for half of these sites with high affinity and for the other half of these sites with significantly lower affinity; 3) antagonists compete for all of these sites with one affinity; and 4) in the presence of disulfide reducing agents, the binding patterns of some agonists and antagonists are changed in accordance with the electrophysiological changes observed under the same conditions with Rana pipiens Sartorius muscle fibers. Our studies indicate that the mechanism of ligand recognition is still functional at equilibrium and is not subject to the presence of an intact membrane environment. Furthermore, the existence of two types of agonist sites at every receptor molecule excludes most of the presently discussed two-state models as the basis for a mechanism of receptor-ligand interaction. To explain sigmoidal dose-response curves, a two-site model is already sufficient.


Electron microscopic evidence for endocytosis of superoxide dismutase by hepatocytes using protein-gold adducts.
Cu, Zn superoxide dismutase, an intracellular, carbohydrate-free protein, was found to bind to hepatocytes and endothelial cells by electron microscopy of liver treated with enzyme-gold conjugates. In cultured hepatocytes, the free enzyme, but not bovine serum albumin, competed for the binding, and at 37 degrees C, the gold-protein complexes were internalized in a typical process of receptor-mediated endocytosis.


Ecophysiology of two solar tracking desert winter annuals : II. Leaf movements, water relations and microclimate.
The seasonal course of water relations in field populations of two leaf solar tracking desert winter annuals was examined. Measurements were made of leaf movements in relation to leaf conductance and water potential. Malvastrum rotundifolium maintained solar tracking movements up to the wilting point of the plant (-4 MPa). Lupinus arizonicus altered its morphology through paraheliotropic leaf movements as leaf water potentials declined to-1.8 MPa. Diurnal patterns of leaf conductance showed marked seasonal trends, with gas exchange activity being restricted to early morning hours as water availability declined. Studies of potted plants showed that L. arizonicus was not able to alter its osmotic potential in response to drought, while M. rotundifolium underwent a 1.86 MPa reduction in osmotic potential. The significance of the two contrasting patterns is discussed in terms of observed plant distribution and origin.


The influence of cholesterol on synaptic fluidity and dopamine uptake.
The present study examined how the synaptic cholesterol/phospholipid ratio influences membrane fluidity and a transmembrane process of the dopaminergic system, dopamine uptake. Fluorescence polarization of DPH was used as a measure of membrane fluidity. The cholesterol content of synaptosomal and synaptic plasma membranes was altered using a lipid transfer protein. The results of the present study demonstrate that the transmembrane process of dopamine uptake may be inhibited by an elevated synaptic cholesterol/phospholipid ratio, which correlates with increased fluorescence polarization. Specifically, a 10-30% increase in the cholesterol/phospholipid ratio resulted in an approximately 20% increase in fluorescence polarization, a 3- to 6-fold increase in Km, a 4-fold increase in Vmax, and a marked decrease in the Vmax/Km (the first order rate constant for dopamine uptake at low substrate concentrations).


Multiple testicular sampling in non-obstructive azoospermia--is it necessary?
Spermatogenesis may be focal in non-obstructive azoospermia. The present study was conducted to determine whether the performance of multiple, rather than a single testicular sample contributes to obtaining spermatozoa in amounts sufficient for fertilization and cryopreservation in non-obstructive, azoospermic patients. Furthermore, the aim was to clarify the significance of location for retrieval from the testis in such cases. Three biopsies were taken from identical locations in 55 testes of 29 men with non-obstructive azoospermia: (i) the rete testis region, ii) the midline, and (iii) the proximal region of the testis. When sperm cells were detected, they were used for intracytoplasmic sperm injection (ICSI), and the remainder were then cryopreserved in as many aliquots as possible (adjusted for ICSI procedure). Spermatozoa were found in 28 testes (50.9%) of 18 men (62.1%). In the testes from which spermatozoa were obtained, they were present in three, two or one locations in 15 (53.6%), five (17.9%) and eight (28.6%) cases respectively. The possibility of finding spermatozoa was not influenced by the location in the testis. Multiple testicular sperm extraction is recommended in cases of non-obstructive azoospermia, since it may enhance diagnostic accuracy of absolute testicular failure and increase the number of sperm cells retrieved.


Implementation of Critical Care Response Teams in Ontario: Impact on the Outcomes of Surgical Patients.
To evaluate whether introduction of CCRTs reduced mortality rates among patients who developed a postoperative complication, also referred to as FTR. CCRTs were introduced to improve patients' postoperative outcomes. Its effect on FTR continues to be actively investigated. We conducted a population-based retrospective cohort study using administrative data from Ontario, Canada. We identified 810,279 patients admitted to hospital for major surgical procedures between January 2004 and December 2014, with a washout period consisting of the 9 months before and after the implementation of CCRTs in January 2007. Difference-in-differences analysis among patients who developed a postoperative complication (n = 148,882) was used to estimate the association between CCRT implementation and FTR before and after CCRT implementation in hospitals that did - versus did not - implement CCRT during the study period. A total of 810,279 patients were included, of whom 148,882 (18.4%) developed a postoperative surgical complication. Among patients who developed a postoperative complication, the overall proportion of FTR was 9.2% (n = 13,659). Among patients in hospitals that introduced CCRT, the RR of FTR was 0.84, [95% confidence interval (CI) 0.78-0.90] after implementation of CCRT, while over the same time period, the RR was 0.85 (95% CI 0.80-0.91) in hospitals that did not implement CCRT. The RR ratio (difference-indifferences) was 0.99 (95% CI 0.89-1.09). Among patients undergoing orthopedic surgery, the RR ratio was 0.84 (95% CI 0.75-0.95). Although implementation of CCRTs in hospitals in Ontario, Canada, did not reduce FTR among all surgical patients having surgery, CCRTs may reduce the risk of FTR among patients having orthopedic surgery.


Power spectrum analysis of myometrial electromyogram and intrauterine pressure changes in the pregnant rhesus monkey in late gestation.
Various methods have been used to quantify myometrial activity in the pregnant rhesus monkey during the last third of gestation. A precise definition of activity must retain the characteristics of epoch duration, amplitude, and the repetitive frequencies of active epochs. We used Fast Fourier Transform and power spectrum analysis to determine whether the patterns of myometrial contractility consist predominantly of contractions or contractures. This analysis allows quantification of activity represented by each of these patterns of activity. We applied these methods to myometrial electromyogram and intrauterine pressure recordings in pregnant rhesus monkeys to characterize the changes in myometrial activity that occur after surgery, after intravenous indomethacin administration, and when food is withdrawn for 48 hours. We conclude that changes in the frequency pattern of myometrial activity provide important information in relation to labor and delivery contractions.


Playback of 22-kHz and 50-kHz ultrasonic vocalizations induces differential c-fos expression in rat brain.
Rodent ultrasonic vocalizations, which serve as sensitive measures in a number of relevant individual and social behaviours, have become increasingly interesting for biopsychological studies on emotion and motivation. Of these, high frequency (50-kHz) ultrasonic vocalizations can index a positive emotional state, and induce approach, whereas low frequency (22-kHz) ultrasonic vocalizations can induce avoidance and may index anxiety, since they are emitted during various unconditioned and conditioned aversive situations. While cholinergic and dopaminergic systems have been implicated, specific neural substrates that sub-serve these vocalization-dependent states remain to be elucidated. Using c-fos immunocytochemistry, we revealed neural activity in brain areas of naïve male Wistar rats in response to playback of 22-kHz and flat and frequency-modulated 50-kHz ultrasonic vocalizations. Presentation of background noise or no acoustic stimulus at all constituted the controls. Playback of 50-kHz ultrasonic vocalizations led to approach behaviour. Acoustically stimulated animals demonstrated differential activation in auditory areas, with a frequency-dependent activation in the auditory cortex. Specific forebrain, thalamic, hypothalamic and brainstem areas were also activated differentially. While 50-kHz playback induced sparse fos-like immunoreactivity in frontal association cortex, nucleus accumbens, thalamic parafascicular and paraventricular nuclei, 22-kHz playback elicited c-fos expression in the perirhinal cortex, amygdalar nuclei and the periaqueductal gray. This study unveils neural substrates that are activated during ultrasonic playback perception, which could sub-serve the affective states elicited by these vocalizations.


A single course of prenatal betamethasone in the rat alters postnatal brain cell proliferation but not apoptosis.
The aim of this study was to determine the effects of a clinically relevant single course of prenatal betamethasone in the rat on growth parameters with particular reference to brain cell proliferation and apoptosis. We report that administration of 170 microg kg-1 betamethasone twice within 4 h to E20 pregnant rats conveys moderate somatic growth retardation. Further, using a measure of brain cell proliferation independent of blood-brain barrier (BBB) permeability, we demonstrate for the first time that betamethasone is chronically anti-proliferative to brain cells without inducing caspase-3-mediated apoptosis. More importantly we show that there is a significant and sexually divergent rebound of neural proliferation which occurs earlier in males than in females and continues until at least 21 days of postnatal life. BBB permeability to [3H]thymidine was significantly increased by steroid treatment re-iterating the fact that tracer studies not correcting for BBB permeability, such as bromodeoxyuridine (BrdU), may be questionable in this type of study. Further, prenatal steroid treatment did not alter postnatal corticosterone levels. In summary we show that prenatal betamethasone conveys significant and long-lasting side effects and that its human clinical application in preterm labour needs more careful consideration as compared to the relative ease with which it is prescribed today.


Novel proline hydroxylase activities in the pneumocandin-producing fungus Glarea lozoyensis responsible for the formation of trans 3- and trans 4-hydroxyproline.
Novel proline 3-hydroxylase (P3H) and proline 4-hydroxylase (P4H) activities that convert free l-proline to both trans 3- and trans 4-hydroxy- l-proline were detected in protein extracts of the anamorphic fungus Glarea lozoyensis. The enzymatic conversion of l-proline to trans 3- and trans 4-hydroxy- l-proline was strictly dependent on alpha-ketoglutarate, ascorbate, and dithiothreitol. Ferrous iron was required for optimal P3H and P4H activity. These substrate and co-factor requirements indicate these enzyme activities belong to the class of 2-oxoglutarate-dependent dioxygenases. Both P3H and P4H were inhibited by zinc and other trace metals. The addition of proline to the fermentation medium resulted in an increase in the specific activity of P4H and a decrease in the synthesis of pneumocandin C(0). Additionally, the synthesis of trans 3- and trans 4-hydroxy- l-proline in vivo was affected differently by the proline concentration in the medium. This result suggested that two enzymes may be responsible for the regio- and stereospecific hydroxylation of l-proline.


Changes in gamma-secretase activity and specificity caused by the introduction of consensus aspartyl protease active motif in Presenilin 1.
Presenilin (PS1 or PS2) is an essential component of the active gamma-secretase complex that liberates the Abeta peptides from amyloid precursor protein (APP). PS1 is regarded as an atypical aspartyl protease harboring two essential aspartic acids in the context of the sequence D257LV and D385FI, respectively, rather than the typical DTG...DTG catalytic motif of classical aspartyl proteases. In the present studies, we introduced the sequence DTG in PS1 at and around the catalytic D257 and D385 residues to generate three PS1 mutants: D257TG, D385TG, and the double-mutant D257TG/D385TG. The effects of these changes on the gamma-secretase activity in the presence or absence of gamma-secretase inhibitors and modulators were investigated. The results showed that PS1 mutants having D385TG robustly enhanced Abeta42 production compared to the wild type (wt), and were more sensitive than wt to inhibition by a classical aspartyl protease transition state mimic, and fenchylamine, a sulfonamide derivative. Unlike wt PS1 and some of its clinical mutants, all three PS1 artificial mutants decreased cleavage of Notch S3-site, suggesting that these artificial mutations may trigger conformational changes at the substrate docking and catalytic site that cause alteration of substrate specificity and inhibition pattern. Consistent with this notion, we have found that NSAID enzymatic inhibitors of COX, known modulators of the gamma-secretase activity, cause PS1 mutants containing D385TG to produce higher levels of both Abeta38 and Abeta42, but to reduce levels of Abeta39, showing a pattern of Abeta formation different from that observed with wild type PS1 and its clinical mutants. This study provides an important structural clue for the rational design of drugs to inhibit processing of APP at the gamma-site without interfering with Notch processing.


Domina (Dom), a new Drosophila member of the FKH/WH gene family, affects morphogenesis and is a suppressor of position-effect variegation.
Domina (Dom) is a novel member of the FKH/WH transcription factor gene family of Drosophila. Two alternatively polyadenylated Dom transcripts of 2.9 and 3.9 kb encode a 719-amino-acid protein with a FKH/WH domain and a putative acidic transactivation domain. Dom is mainly expressed in the central and peripheral nervous system. Homozygous mutants show rough eyes, irregular arrangement of bristles, extended wings, defective posterior wing margins, and a severely diminished vitality and fertility. Heterozygous Dom flies are morphologically wild type but show suppression of position-effect variegation. Consistently with this chromatin effect DOM protein is accumulated in the chromocenter and, as expected from a transcription factor, is found at specific euchromatic loci. Sequence comparison suggests that DOM of Drosophila is homologous to the chordate WHN proteins. The chromatin modifying capability of DOM is probably based on the FKH/WH domain, which shows a remarkable structural similarity to the winged-helix structures of H1 and the central globular domain of H5.


The impact of detectives' manner of questioning on rape victims' disclosure.
Research has documented that few reported rapes are prosecuted by the legal system. The purpose of this study is to explain how the interactions between victims and detectives can strengthen or weaken the investigation itself. Twenty rape victims were interviewed to examine how law enforcement detectives' manner of questioning affects rape victims' level of disclosure. Using qualitative methodology, the results show that the detectives' manner of questioning can play a role in victims' disclosure. Detectives using a gentle manner of questioning with victims can help produce stronger victim statements and thus build stronger cases for prosecution.


Investigation of flexible hose insertion forces and selected factors.
Flexible hoses are widely used in automobiles. Understanding the force required to insert a hose, independent of worker variation, is important for engineers, designers and health professionals. Various parameters affect the insertion forces including the interference between the hose and flange. Also, it has been observed that workers often add motions during the insertion process, including rotation and oscillation. This paper describes the effects of these parameters on axial insertion forces. A programmable mill was used to isolate the effects of four variables on hose insertion force: fit, insertion speed, rotational velocity and oscillation magnitude. The results show that the magnitude of the interference of the 19 mm inside diameters (ID) heater hose and the flange, and the insertion technique affect the insertion forces; resulting in a range of insertion forces from 11.1+/-0.2 to 128.4+/-11.3N.


Antiviral death punch by ADP-ribosylating bacterial toxins.
Toxin-antitoxin systems can defend bacteria against phages by shutting down infected cells, but the links between their molecular mechanisms and biological functions have remained underexplored. LeRoux et al. now show how the DNA-targeting ADP-ribosylation activity of DarTG impairs phage replication but is overcome by dedicated viral inhibitors and evolved tolerance.


[Use of Spanish proposals for measuring social class in health sciences].
Recognizing the need for a consensus tool to measure social class in health sciences, in 1995 the Spanish Society of Epidemiology (Sociedad Española de Epidemiología [SEE]) made a proposal for such a measure. The aim of this article was to explore the bibliometric impact of the SEE's proposal. Articles citing the SEE's report and those citing articles published as a result of the report were identified using Google Scholar and Scopus. The information extracted from these articles consisted of year, nationality and impact factor of the journal, self-citation, information source, subject area, city of the first author, type of publication, study design, use of social class as a variable, number of social class categories, and the use of education or occupation to determine social class. The number of citations progressively increased. Citations also showed heterogeneity in the subject. Generally, citations were found in original articles and articles with a cross sectional design, were published in journals with impact factor and were by researchers working in Barcelona or Madrid. The SEE's proposal has proved to be an increasingly useful tool, applicable in different contexts. As suggested by the SEE, this proposal may need to be updated.


Knockdown of long non-coding RNA MALAT1 increases the blood-tumor barrier permeability by up-regulating miR-140.
The blood-tumor barrier (BTB) forms a major obstacle in brain tumor therapy by preventing the delivery of sufficient quantities of therapeutic drugs. Long non-coding RNAs (lncRNAs) play important roles in both normal development and diseases including cancer. Here, we elucidated the expression of lncRNA metastasis associated lung adenocarcinoma transcript 1 (MALAT1) and defined its functional role in the regulation of BTB function as well as its possible molecular mechanisms. Our results proved that MALAT1 expression was up-regulated in brain microvessels of human glioma and glioma endothelial cells (GECs) which were obtained by co-culturing endothelial cells with glioma cells. Functionally, knockdown of MALAT1 resulted in an impairment and increased the permeability of BTB as well as decreased the expression of ZO-1, occludin and claudin-5 in GECs. Further, there was reciprocal repression between MALAT1 and miR-140, and miR-140 mediated the effects that MALAT1 knockdown exerted. Mechanistic investigations defined that nuclear factor YA (NFYA), a CCAAT box-binding transcription factor, was a direct and functional downstream target of miR-140, which was involved in the MALAT1 knockdown induced regulation of BTB function. Furthermore, NFYA could up-regulate the promoter activities and bind to the promoters of ZO-1, occludin and claudin-5 in GECs. Taken together, we have demonstrated the fact that knockdown of MALAT1 resulted in the increased permeability of BTB, which might contribute to establishing potential therapeutic strategies for human gliomas.


Development and comparison of three cell-based potency assays for anti-respiratory syncytial virus monoclonal antibody.
There is an increasing demand for monoclonal antibody (mAb) therapies to confer passive immunity against viral diseases. Respiratory syncytial virus (RSV) is the most common cause of bronchiolitis, lower respiratory tract infections, and hospitalization in infants. Currently, there is no RSV vaccine but a humanized mAb available for high risk infants. MK-1654 is a fully human mAb with YTE mutation in the fragment crystallizable (Fc) region to extend the half-life in circulation. It binds to a highly conserved epitope of RSV Fusion protein with high affinity and neutralizes RSV infection. A functional cell-based assay is a regulatory requirement for clinical development, commercial release, and stability testing of MK-1654. In this study, we have evaluated three RSV neutralization assays to test the potency of MK-1654, including an imaging-based virus reduction neutralization test (VRNT) and two reporter virus-based assays (RSV-GFP and RSV-NLucP). All three methods showed good dose response curves of MK-1654 with similar EC50 values. RSV-NLucP method was chosen for further development because it is simple and can be easily adapted to quality control testing laboratories. After optimization, the RSV-NLucP assay was pre-qualified with good linearity, relative accuracy, intermediate precision, and specificity, therefore suitable for a cell-based potency assay.


PD-1 Blockade Boosts Radiofrequency Ablation-Elicited Adaptive Immune Responses against Tumor.
Radiofrequency ablation (RFA) has been shown to elicit tumor-specific T-cell immune responses, but is not sufficient to prevent cancer progression. Here, we investigated immune-suppressive mechanisms limiting the efficacy of RFA. We performed a retrospective case-controlled study on patients with synchronous colorectal cancer liver metastases who had received primary tumor resection with or without preoperative RFA for liver metastases. Tumor-infiltrating T cells and tumoral PD-L1 expression in human colorectal cancer tissues were analyzed by immunohistochemistry. T-cell immune responses and PD-1/PD-L1 expression were also characterized in an RFA mouse model. In addition, the combined effect of RAF and PD-1 blockade was evaluated in the mouse RFA model. We found that RFA treatment of liver metastases increased not only T-cell infiltration, but also PD-L1 expression in primary human colorectal tumors. Using mouse tumor models, we demonstrated that RFA treatment of one tumor initially enhanced a strong T-cell-mediated immune response in tumor. Nevertheless, tumor quickly overcame the immune responses by inhibiting the function of CD8(+) and CD4(+) T cells, driving a shift to higher regulatory T-cell to Teff ratio, and upregulating PD-L1/PD-1 expression. Furthermore, we established that the combined therapy of RFA and anti-PD-1 antibodies significantly enhanced T-cell immune responses, resulting in stronger antitumor immunity and prolonged survival. The PD-L1-PD-1 axis plays a critical role in dampening RFA-induced antitumor immune responses, and this study provides a strong rationale for combining RFA and the PD-L1/PD-1 blockade in the clinical setting.


Evolutionary implications of SARS-CoV-2 vaccination for the future design of vaccination strategies.
Once the first SARS-CoV-2 vaccine became available, mass vaccination was the main pillar of the public health response to the COVID-19 pandemic. It was very effective in reducing hospitalizations and deaths. Here, we discuss the possibility that mass vaccination might accelerate SARS-CoV-2 evolution in antibody-binding regions compared to natural infection at the population level. Using the evidence of strong genetic variation in antibody-binding regions and taking advantage of the similarity between the envelope proteins of SARS-CoV-2 and influenza, we assume that immune selection pressure acting on these regions of the two viruses is similar. We discuss the consequences of this assumption for SARS-CoV-2 evolution in light of mathematical models developed previously for influenza. We further outline the implications of this phenomenon, if our assumptions are confirmed, for the future design of SARS-CoV-2 vaccination strategies.


PEGylation as a strategy for improving nanoparticle-based drug and gene delivery.
Coating the surface of nanoparticles with polyethylene glycol (PEG), or "PEGylation", is a commonly used approach for improving the efficiency of drug and gene delivery to target cells and tissues. Building from the success of PEGylating proteins to improve systemic circulation time and decrease immunogenicity, the impact of PEG coatings on the fate of systemically administered nanoparticle formulations has, and continues to be, widely studied. PEG coatings on nanoparticles shield the surface from aggregation, opsonization, and phagocytosis, prolonging systemic circulation time. Here, we briefly describe the history of the development of PEGylated nanoparticle formulations for systemic administration, including how factors such as PEG molecular weight, PEG surface density, nanoparticle core properties, and repeated administration impact circulation time. A less frequently discussed topic, we then describe how PEG coatings on nanoparticles have also been utilized for overcoming various biological barriers to efficient drug and gene delivery associated with other modes of administration, ranging from gastrointestinal to ocular. Finally, we describe both methods for PEGylating nanoparticles and methods for characterizing PEG surface density, a key factor in the effectiveness of the PEG surface coating for improving drug and gene delivery.


Amino acid sequence of phospholipase A2-alpha from the venom of Crotalus adamanteus. A new classification of phospholipases A2 based upon structural determinants.
The complete amino acid sequence of Crotalus adamanteus venom phospholipase A2-alpha has been determined by analysis of the five tryptic peptides from the citraconylated, reduced, and S-[14C]carboxamidomethylated enzyme. Earlier studies (Tsao, F. H. C., Keim, P. S., and Heinrikson, R. L. (1975) Arch. Biochem. Biophys. 167, 706) provided the information necessary to align the tryptic fragments so that secondary cleavage procedures to establish overlaps were unnecessary. The subunit in the phospholipase A2-alpha dimer is a single polypeptide chain containing 122 amino acids and seven disulfide bonds. The histidine residue implicated in the active site of mammalian phospholipases is at position 47 in the C. adamanteus enzyme and is located in a domain of the molecule which is highly homologous in sequence with corresponding regions of phospholipases from a variety of venom and pancreatic sources. Comparative sequence analysis has revealed insights with regard to the function and evolution of phospholipases A2. Primary structural relationships observed among the snake venom enzymes parallel the phylogenetic classification of the venomous reptiles from which they were derived. It is proposed that phospholipases A2 of this general type be divided into two groups depending upon the presence or absence of distinctive structural features elucidated in this study.


Detrimental effects of beta-adrenergic stimulation on beta-adrenoceptors and microtubules in the heart.
Increased plasma catecholamines - in particular, excessive beta-adrenoceptor activation in chronic heart failure - may easily desensitize the beta-adrenoceptors as well as the postreceptor signal transductions. Since these detrimental changes in the failing heart could be reversible, administration of low-dose beta-blocker, which minimizes the negative inotropic effects, may be effective in attenuating the harmful effects of sympathetic nerve activation. Beta-adrenoceptor stimulation may also produce microtubule disruptions of the cell either through direct action or through an increase in heart rate. Treatment with beta-blockers could attenuate Ca overload by slowing the heart rate and may be useful as a protection from the structural disintegration of the cell. Thus, to clarify the underlying mechanisms of beta-blocker therapy for chronic heart failure, we have to consider not only to the functional aspects but also to the structural changes of the cells.


Continuous succinic acid production from xylose by Actinobacillus succinogenes.
Continuous, anaerobic fermentations of D-xylose were performed by Actinobacillus succinogenes 130Z in a custom, biofilm reactor at dilution rates of 0.05, 0.10 and 0.30 h(-1). Succinic acid yields on xylose (0.55-0.68 g g(-1)), titres (10.9-29.4 g L(-1)) and productivities (1.5-3.4 g L(-1) h(-1)) were lower than those of a previous study on glucose, but product ratios (succinic acid/acetic acid = 3.0-5.0 g g(-1)) and carbohydrate consumption rates were similar. Also, mass balance closures on xylose were up to 18.2 % lower than those on glucose. A modified HPLC method revealed pyruvic acid excretion at appreciable concentrations (1.2-1.9 g L(-1)) which improved the mass balance closure by up to 16.8 %. Furthermore, redox balances based on the accounted xylose consumed and the excreted metabolites, indicated an overproduction of reducing power. The oxidative pentose phosphate pathway was shown to be a plausible source of the additional reducing power.


Advances in oral peptide drug nanoparticles for diabetes mellitus treatment.
Peptide drugs play an important role in diabetes mellitus treatment. Oral administration of peptide drugs is a promising strategy for diabetes mellitus because of its convenience and high patient compliance compared to parenteral administration routes. However, there are a series of formidable unfavorable conditions present in the gastrointestinal (GI) tract after oral administration, which result in the low oral bioavailability of these peptide drugs. To overcome these challenges, various nanoparticles (NPs) have been developed to improve the oral absorption of peptide drugs due to their unique in vivo properties and high design flexibility. This review discusses the unfavorable conditions present in the GI tract and provides the corresponding strategies to overcome these challenges. The review provides a comprehensive overview on the NPs that have been constructed for oral peptide drug delivery in diabetes mellitus treatment. Finally, we will discuss the rational application and give some suggestions that can be utilized for the development of oral peptide drug NPs. Our aim is to provide a systemic and comprehensive review of oral peptide drug NPs that can overcome the challenges in GI tract for efficient treatment of diabetes mellitus.


Clomipramine in the treatment of agoraphobic inpatients resistant to behavioral therapy.
Both behavior-modification methods and antidepressants have proved to be effective in the treatment of agoraphobia. The authors examined the effects of clomipramine on agoraphobia in patients who failed to respond to exposure-based behavioral treatment. Eighteen patients with panic disorder with agoraphobia who had not responded to previous inpatient behavioral treatment were recruited to a 12-week, placebo-controlled, double-blind crossover study of clomipramine, at top doses of 150 mg/day for 3 weeks. The patients were assessed on measures of phobic avoidance, agoraphobic cognitions, panic, state and trait anxiety, subjective anxiety, and depression. One patient dropped out of the study after 6 weeks. On most outcome measures, the 17 study completers had significantly (p < .05) lower symptom scores at posttest in the active drug period than at posttest in the placebo period; however, the clinical gains were modest. The short-term efficacy of clomipramine for agoraphobic patients who failed to respond lastingly to behavioral treatment was demonstrated. It remains to be shown that clomipramine can lead to clinically significant and lasting benefits in these patients.


NADPH: nitroblue tetrazolium reductase found in plasma membrane of human neutrophil.
After phorbol 12-myristate 13-acetate (PMA) stimulation the increase of NADPH:nitroblue tetrazolium reductase activity in the plasma membrane almost corresponded with the stimulated activity of respiratory burst oxidase. Solubilization of plasma membranes from PMA-activated neutrophils with n-octyl glucoside resulted in high recoveries of the two enzymatic activities. When solubilized plasma membrane was subjected to non-denaturing polyacrylamide gel electrophoresis in the presence of 35 mM n-octyl glucoside, we could see three major bands stained with NADPH-dependent nitroblue reductase activity giving molecular masses of approx. 95, 45 and 40 kDa, respectively. Activity was specific for NADPH but not for NADH. These bands also stained weakly in the plasma membranes obtained from resting cells. The activities for NADPH oxidase and nitroblue tetrazolium reductase were found to elute as a very similar protein peak on an anion-exchange HPLC, at about 0.32 M KCl. This elution peak also contains 45 and 40 kDa proteins showing NADPH:nitroblue tetrazolium reductase activity.


Anesthetic considerations for airway stenting in adult patients.
The attributable morbidity from central airway obstruction is significant. Airway stenting provides a therapeutic option to manage these complex lesions. This article focuses on the relevant anesthetic considerations of airway stenting in adult patients.


GABA promotes the competitive selection of dendritic spines by controlling local Ca2+ signaling.
Activity-dependent competition of synapses plays a key role in neural organization and is often promoted by GABA; however, its cellular bases are poorly understood. Excitatory synapses of cortical pyramidal neurons are formed on small protrusions known as dendritic spines, which exhibit structural plasticity. We used two-color uncaging of glutamate and GABA in rat hippocampal CA1 pyramidal neurons and found that spine shrinkage and elimination were markedly promoted by the activation of GABAA receptors shortly before action potentials. GABAergic inhibition suppressed bulk increases in cytosolic Ca(2+) concentrations, whereas it preserved the Ca(2+) nanodomains generated by NMDA-type receptors, both of which were necessary for spine shrinkage. Unlike spine enlargement, spine shrinkage spread to neighboring spines (<15 μm) and competed with their enlargement, and this process involved the actin-depolymerizing factor ADF/cofilin. Thus, GABAergic inhibition directly suppresses local dendritic Ca(2+) transients and strongly promotes the competitive selection of dendritic spines.


In vivo effects of lipopolysaccharide on hepatic free-NAD(P)(+)-linked redox states and cytosolic phosphorylation potential in 48-hour-fasted rats.
This study was performed to determine the magnitude and time of onset of in vivo changes in hepatic bioenergetics in response to a sublethal dose of lipopolysaccharide (LPS), a bacterial endotoxin. Male rats (48-hour-fasted) were administered an intraperitoneal injection of LPS (5 mg/kg body weight) or vehicle alone, and the livers were freeze-clamped 5, 30, or 180 minutes or 24 hours later. Liver tissue was extracted with perchloric acid, and the metabolites necessary to calculate NAD(+)- and NADP(+)-linked redox states and the cytosolic phosphorylation potential were measured. There was no significant difference in hepatic cytosolic phosphorylation potential between LPS and control groups at any of the times investigated. This indicated that the ability of the liver to synthesize adenosine triphosphate (ATP) was not compromised under the conditions of the study. No changes in hepatic redox states were observed 5 or 30 minutes after LPS treatment. Three hours after LPS treatment, hepatic cytosolic and mitochondrial free-[NAD+]/[NADH] redox states and the cytosolic free-[NADP+]/[NADPH] redox state were more oxidized. By 24 hours, only NAD(+)-linked redox states were more oxidized than the time-matched controls. Hepatic urea content was elevated at both 3 and 24 hours, compatible with an increased rate of urea synthesis as a consequence of increased amino acid metabolism, whereas hepatic beta-hydroxybutyrate and total ketone bodies were decreased 24 hours after LPS treatment, indicating decreased hepatic ketogenesis.(ABSTRACT TRUNCATED AT 250 WORDS)


Stress, the adrenergic hypothalamovagal pathway, and the aetiology of chronic duodenal ulceration.
In the rat, stress activates the hypothalamus causing central adrenergic discharge with delivery of an alpha-adrenergic stimulus by the adrenergic hypothalamovagal pathway to the stomach. This stimulus controls intragastric blood flow and 5-HT release. In rats stressed pharmacologically by administration of intraperitoneal reserpine (0.1 mg/kg), the stimulated pathway releases gastric 5-HT which in turn significantly increases gastric acid secretion. Administration of intramuscular reserpine (0.1 mg/kg) every 24 hr for six weeks significantly (p less than 0.001 by Wilcoxon matched pairs signed-rank test) increased the basal acid output of the rat stomach (12 mumol/h +/- 0.5 vs 38.1 mumol/h +/- 2.4, mean +/- SEM) and produced duodenal ulceration in 83.3% of animals. Vagotomy completely protected the rat stomach against these effects. The data show that prolonged stress in the rat stimulates gastric acid secretion and produces duodenal ulceration.


Water loss and malate fluctuations during the day for plants in the southern Namib desert.
The daily course of transpiration for 12 different plants growing in the southern Namib desert was investigated. Sclerophyllous species with C3 photosynthesis were the most effective water savers followed by succulents exhibiting CAM, while C3 pathway-succulents lose as much water as malacophyllous species. The different species showed either one or two peaked patterns of both transpiration and leaf conductance. With C3 plants the most common pattern is a single morning peak in leaf conductance followed by decreases in conductance over the remainder on the day. With CAM succulents leaf conductance is high in the morning, shows a pronounced midday depression and increases in the early afternoon when the malate pool is depleted, but malate consumption did not start earlier than 4 h after dawn. Seven of nine investigated C3 plants had rather high contents of malate, up to 180 μmol·g-1 dry matter. In these plants the malate content increased with increasing leaf conductance and disappeared when leaf conductance declined in the evening, indicating that malate was synthesized during photosynthesis.


Polymorphism and desolvation of flupirtine maleate.
Crystallizates of the analgesic agent flupirtine maleate (Katadolon; ASTA Medica, Dresden, Germany) obtained from isopropanol are examined by X-ray diffractometry, polarization microscopy and thermoanalysis. Depending on the crystallizing conditions, the modifications A and B as well as an isopropanol solvate are observed. The inversion temperature A-->B of the enantiotropic modifications is 164 degrees C (differential scanning calorimetry (DSC) onset). During thermal desolvation, modification B is formed well below the inversion temperature. In concentrated isopropanol suspensions, the solvate and modification B are rapidly transformed into modification A. It is shown how phase-pure products consisting of modification A, which is better wettable with water and stable at room temperature, can be obtained.


Two-Color Total Internal Reflection Fluorescence Microscopy of Exocytosis in Endocrine Cells.
We describe a comprehensive method for imaging and analysis of local protein dynamics at single sites of exocytosis in living cultured endocrine cells. This method is well suited to quantitatively map the complex dynamics of individual molecules at single sites of vesicle fusion in live cells.


Attention-Based Deep Learning System for Classification of Breast Lesions-Multimodal, Weakly Supervised Approach.
Breast cancer is the most frequent female cancer, with a considerable disease burden and high mortality. Early diagnosis with screening mammography might be facilitated by automated systems supported by deep learning artificial intelligence. We propose a model based on a weakly supervised Clustering-constrained Attention Multiple Instance Learning (CLAM) classifier able to train under data scarcity effectively. We used a private dataset with 1174 non-cancer and 794 cancer images labelled at the image level with pathological ground truth confirmation. We used feature extractors (ResNet-18, ResNet-34, ResNet-50 and EfficientNet-B0) pre-trained on ImageNet. The best results were achieved with multimodal-view classification using both CC and MLO images simultaneously, resized by half, with a patch size of 224 px and an overlap of 0.25. It resulted in AUC-ROC = 0.896 ± 0.017, F1-score 81.8 ± 3.2, accuracy 81.6 ± 3.2, precision 82.4 ± 3.3, and recall 81.6 ± 3.2. Evaluation with the Chinese Mammography Database, with 5-fold cross-validation, patient-wise breakdowns, and transfer learning, resulted in AUC-ROC 0.848 ± 0.015, F1-score 78.6 ± 2.0, accuracy 78.4 ± 1.9, precision 78.8 ± 2.0, and recall 78.4 ± 1.9. The CLAM algorithm's attentional maps indicate the features most relevant to the algorithm in the images. Our approach was more effective than in many other studies, allowing for some explainability and identifying erroneous predictions based on the wrong premises.


Workplace inequality is associated with status-signaling expenditure.
Regional inequality is known to magnify sensitivity to social rank. This, in turn, is shown to increase people's propensity to acquire luxury goods as a means to elevate their perceived social status. Yet existing research has focused on broad, aggregated datasets, and little is known about how individual-level measures of income interact with inequality within peer groups to affect status signaling. Using detailed financial transaction data, we construct 32,008 workplace peer groups and explore the longitudinal spending and salary data associated with 683,677 individuals. These data reveal links between people's status spending, their absolute salary, salary rank within their workplace peer group, and the inequality of their workplace salary distribution. Status-signaling luxury spending is found to be greatest among those who have higher salaries, whose workplaces exhibit higher inequality, and who occupy a lower rank position within the workplace. We propose that low-rank individuals in unequal workplaces suffer status anxiety and, if they can afford it, spend to signal higher status.


Family, healthcare professional, and societal preferences for the treatment of infantile spinal muscular atrophy: A discrete choice experiment.
To understand the factors that most influence decision-making in the treatment of infantile spinal muscular atrophy (SMA). A discrete choice experiment was conducted among parents of people with SMA (parents), healthcare professionals (HCPs), and members of the Australian general population (GenPop). Respondents were asked to accept/reject treatment for an infant newly diagnosed with SMA in eight hypothetical scenarios, characterized by different combinations of the attributes of the treatment offered. The results were analyzed using probability analysis. Completed responses were provided from 1113 individuals (1024 GenPop, 21 parents, 68 HCPs). Respondents were more likely to accept treatments that improved functioning and mobility. Treatments with higher costs, invasive delivery, and risks of adverse events were accepted less often. Cost most affected treatment choices by HCPs and GenPop, while change in mobility and mode of administration were most influential for parents. These results highlight the importance of understanding value for money and clinical impact in affecting treatment choice, which are crucial for effective planning of healthcare and the successful implementation of treatment programmes for SMA. What this paper adds Spinal muscular atrophy (SMA) treatments with a higher chance of improving functioning and mobility are preferred by the general population, parents, and healthcare professionals. Treatments with higher costs, invasive delivery, and risk of adverse events are less preferred. Willingness to pay for SMA treatments increases with impact on functioning.