Training adults with acquired brain injury how to help-seek when wayfinding: an understudied critical life skill.
The objective of this study was to investigate the efficacy of a group treatment protocol called NICE (Noticing you have a problem, Identifying the information you need for help, Compensatory strategies, Evaluating progress) to train help-seeking when wayfinding for individuals with acquired brain injury (ABI). Seven participants completed the NICE group treatment in an outpatient rehabilitation department at a university medical centre. A single subject multiple baseline design was employed to evaluate the efficacy of the NICE group treatment. The Social Behaviour Rating Scale and the Executive Function Route-Finding Task- Revised were repeated measures used to evaluate potential changes in help-seeking and wayfinding. Secondary outcome measures included pre- and post-treatment evaluation of social problem solving and social cognition. Results revealed that all participants improved on measures of help-seeking and wayfinding. Patterns of improvement and implications for rehabilitation are discussed. This is the first experimental study to evaluate the treatment of help-seeking behaviours and discuss its application to wayfinding in adults with ABI. Preliminary evidence supports further investigation of the NICE group treatment protocol.


Designing a better theranostic nanocarrier for cancer applications.
Nanocarriers show incredible potential in theranostic applications as they offer diagnostic capabilities along with the ability to encapsulate and protect drugs from degradation, be functionalized with targeting moieties and be designed with controlled release mechanisms. Most clinically approved nanocarrier drugs are liposomal formulations. As such, considerable research has been directed towards designing liposomal carriers that can release their payloads via exogenous or endogenous triggers. For triggered release to effectively increase drug bioavailability, nanocarriers must first accumulate at the tumor site via the enhanced retention and permeability effect. It has been demonstrated in the chicken embryo chorioallantoic membrane and murine xenografted models that nanoparticle surface charge and geometry, with respect to vascular endothelium fenestration size, drive this accumulation in angiogenic tissue.


Relationship between serum uric acid levels and osteoporosis.
Osteoporosis (OP) is a systemic bone disease in which bone density and quality decrease and bone fragility increases due to a variety of causes, making it prone to fractures. The development of OP is closely related to oxidative stress. Uric acid (UA) is the end product of purine metabolism in the human body. Extracellular UA has antioxidant properties and is thought to have a protective effect on bone metabolism. However, the process of UA degradation can lead to intracellular oxidative stress, which together with UA-induced inflammatory factors, leads to increased bone destruction. In addition, UA can inhibit vitamin D production, resulting in secondary hyperparathyroidism and further exacerbating UA-associated bone loss. This review summarizes the relationship between serum UA levels and bone mineral density, bone turnover markers, and so on, in the hope of providing new insights into the pathogenesis and treatment of OP.


Lymphoreticular aggregates in the newborn lung and their relation to intrauterine INFECTION and neurospinal dysraphism.
A survey has been carried out of lymphoreticular aggregates in the lungs of 200 children, stillborns and dying within 15 days of birth and in a further 21 children with anencephalus, and controls. Lymphoreticular aggregates occur in response to amniotic infection and are found in the 'intrauterine infection syndrome'. They are not found in children showing early hyaline membranes. The time interval required for the production of lymphoreticular aggregates in the human child appears to be at lease 48 h and is probably 72 h before the production of well-formed aggregates and this probably applies to both the intrauterine and the postnatal state. There is a significant increase in the number of lungs showing lymphoreticular aggregates at birth in children in whom there is neurospinal dysraphism. In such children there is necrosis of nervous tissue into the amniotic cavity and it would seem likely that this material has been antigenic to the lung in utero.


CTLA-4 gene polymorphism contributes to the genetic risk for latent autoimmune diabetes in adults.
Latent autoimmune diabetes in adults (LADA) is identified by the presence of GAD65 autoantibodies in diabetic patients who do not require insulin treatment for at least six months after the diagnosis. Previous studies have shown that the risk for LADA, similarly to type 1 diabetes mellitus (T1DM), is increased in subjects carrying the HLA-DRB1*03-DQA1*0501-DQB1*0201 and/or HLA-DRB1*04-DQA1*0301-DQB1*0302 haplotypes. In the present study, we investigated the association between LADA and the CTLA-4 A/G polymorphism, another gene polymorphism associated with T1DM and other autoimmune diseases. The heterozygous A/G genotype was significantly more frequent among 80 LADA (69%) than among 85 healthy subjects of similar age and geographical provenience (47%) (OR = 2.47, corrected P = 0.023). Conversely, the homozygous A/A genotype was significantly less frequent in LADA subjects than in healthy controls (26% vs. 47%, OR = 0.4, corrected P = 0.028). The results of our study show that LADA is positively associated with the CTLA-4 A/G genotype, similarly to T1DM, thus providing further supporting evidence of the autoimmune origin of this form of diabetes mellitus of the adult.


Antiarrhythmic use of cibenzoline, a new class 1 antiarrhythmic agent with class 3 and 4 properties, in patients with recurrent ventricular tachycardia.
Cibenzoline is a new antiarrhythmic agent with class 1 properties, and additional class 3 and 4 effects. We treated 28 patients with drug-refractory and recurrent ventricular tachycardia with up to 700 mg/day cibenzoline for periods up to 5.5 months. Cibenzoline prevented the recurrence of ventricular tachycardia in five patients (18%). In three patients (11%) the arrhythmia may have been worsened, in 23 patients (82%) cibenzoline was ineffective. Cibenzoline increased the PR interval by 18% and the QRS duration by 33%; the effect on the QT was variable and the corrected QT interval did not change significantly. Side-effects were observed in 21% of patients. We conclude that cibenzoline does not appear to be superior to conventional class 1 antiarrhythmic agents and that it cannot be recommended for general use in patients with ventricular tachycardia. Additional pharmacokinetic and electrophysiologic studies are required before cibenzoline is used in outpatients with severe ventricular arrhythmias.


An Overview of Two-Component Signal Transduction Systems Implicated in Extra-Intestinal Pathogenic E. coli Infections.
Extra-intestinal pathogenic E. coli (ExPEC) infections are common in mammals and birds. The predominant ExPEC types are avian pathogenic E. coli (APEC), neonatal meningitis causing E. coli/meningitis associated E. coli (NMEC/MAEC), and uropathogenic E. coli (UPEC). Many reviews have described current knowledge on ExPEC infection strategies and virulence factors, especially for UPEC. However, surprisingly little has been reported on the regulatory modules that have been identified as critical in ExPEC pathogenesis. Two-component systems (TCSs) comprise the predominant method by which bacteria respond to changing environments and play significant roles in modulating bacterial fitness in diverse niches. Recent studies have highlighted the potential of manipulating signal transduction systems as a means to chemically re-wire bacterial pathogens, thereby reducing selective pressure and avoiding the emergence of antibiotic resistance. This review begins by providing a brief introduction to characterized infection strategies and common virulence factors among APEC, NMEC, and UPEC and continues with a comprehensive overview of two-component signal transduction networks that have been shown to influence ExPEC pathogenesis.


The role of genetic analysis for predicting outcome of rheumatoid arthritis.
Rheumatoid Arthritis (RA) varies from a mild to a severe, unremitting illness characterized by uncontrolled inflammation with consequent damage to cartilage and bone of joints. Individualized therapeutic approaches based on likely outcome would facilitate a personalized therapeutic approach. Areas covered: Genetics is known to contribute a significant component of the variability in RA outcome, estimated at 45-60%. A number of candidate gene studies have been associated with variability in radiologically assessed joint damage; however a more comprehensive genome wide analysis is required to more fully characterize the genetic basis of RA severity. Expert commentary: Genetic profiling of patient presenting with RA has the potential to aid stratification based on predicted prognosis, this would inform the clinical development of a personalized therapeutic approach. It will also result in the identification of novel mediators of tissue damage in RA.


Cloning of a ribonucleotide reductase gene of the herpes simplex virus type 2 strain G.
The ribonucleotide reductase (RR) 2 gene of the HSV-2 strain G was cloned, sequenced, and expressed in an E. coli cell. The RR2 gene was located on the PstI 2.4 kb fragment, which was cloned and sequenced. The ORF of the gene was 1,011 bp and its termination codon was TAG; also, the CATATAA sequence was present in the promoter of the RR2 gene. A Poly A signal sequence (AATAAA) was found in the 3'-noncoding region. The RR2 proteins that were produced in the E. coli and Vero cells were confirmed using a Western blot analysis. SDS-PAGE revealed that the molecular weights of the fusion-RR2 that was produced in the E. coli cells were approximately 24 kDa and 38 kDa in the Vero cells. The RR2 proteins were soluble. The differences in the molecular weights might be due to modifications in the Vero cells.


Correction of infraorbital dark circles using collagenase-digested fat cell grafts.
A number of factors, including visibility of the orbicularis oculi muscle through thin skin, can cause dark circles around the eyes. Fat grafts have been used to augment the lower eyelid skin to correct dark circles, but irregularities caused by leaving visible lumps of the fat can occur. We used collagenase-digested fat cell grafts to correct these deformities. Fat was aspirated from the medial thigh using the Coleman method. After centrifugation, the middle fat layer was segregated, digested using Clostridium histolyticum-derived type II collagenase, and mixed with Hartman's solution. The grafts were injected into the infradermal layer using blunt 22-G needles. Eighty-two patients were evaluated for improvement using photographic evidence; 55 (67.1%) showed improvement, 23 (28%) showed no difference, and four (4.9%) had worsened from preoperative findings. There were no visible lumps of fat, contour irregularities, or fat necroses. Postoperative edema and ecchymosis were minimal. Collagenase-digested fat cell grafts provide another option for correcting dark circles by augmenting thin skin. Further histologic evaluation of the grafted collagenase-digested fat cells is recommended.


[Passive microhemagglutination reaction to Treponema pallidum in the cerebrospinal fluid in the diagnosis of neurosyphilis].
A quantitative micro-hemagglutination test for antibodies to Treponema pallidum (MHA-TP) was evaluated in 25 cerebrospinal fluid (CSF) samples obtained from patients with neurosyphilis (NS group) and in 7 CSF samples of patients with reactive serologic tests for syphilis (STS+). These data were compared to treponemal and nontreponemal tests. The MHA-TP was reactive in all of the 25 NS group samples, the FTA-Abs and the complement fixation of Wassermann (CFW) were in 24 and the VDRL in only 9. In the 7 STS+ samples (STS+ group), the MHA-TP was reactive in 6, the FTA-Abs in all of them and the nontreponemal tests were nonreactive. Results analyses support conclusion the clinical diagnosis of NS must be complemented by cytoproteic dual and immunological treponemal and nontreponemal assays in CSF. The MHA-TP test was as sensitive as FTA-Abs and required less technical and interpretative skills, contributing in association to CFW to NS diagnosis.


A microfluidics assisted porous silicon array for optical label-free biochemical sensing.
A porous silicon (PSi) based microarray has been integrated with a microfluidic system, as a proof of concept device for the optical monitoring of selective label-free DNA-DNA interaction. A 4 × 4 square matrix of PSi one dimensional photonic crystals, each one of 200 μm diameter and spaced by 600 μm, has been sealed by a polydimethylsiloxane (PDMS) channels circuit. The PSi optical microarray elements have been functionalized by DNA single strands after sealing: the microfluidic circuit allows to reduce significantly the biologicals and chemicals consumption, and also the incubation time with respect to a not integrated device. Theoretical calculations, based on finite element method, taking into account molecular interactions, are in good agreement with the experimental results, and the developed numerical model can be used for device optimization. The functionalization process and the interaction between DNA probe and target has been monitored by spectroscopic reflectometry for each PSi element in the microchannels.


Species Specificity on Interaction between IgE and FcεRI.
Allergic diseases are one of the most prevalent diseases at present, it is imperative to understanding the pathophysiology and treatment strategies for allergic diseases. In this process, the binding of IgE and FcεRI on effector cells plays a critical role in triggering allergic reactions. However, the species specificity of the interaction between IgE and FcεRI has not been clearly explained. This review described the characteristics and the interaction mechanism in the allergic reaction of IgE and FcεRI and summarized the species specificity between IgE and FcεRI.


Rh(III)-Catalyzed C-H Activation of Benzoylacetonitriles and Tandem Cyclization with Diazo Compounds to Substituted Benzo[ de]chromenes.
Rh (III)-catalyzed C-H activation of benzoylacetonitriles in coupling with diazo compounds was developed to synthesize diversified substituted benzo[ de]chromenes via a formal (4 + 2) cycloaddition with a diazo compound and subsequent tandem (4 + 2) cycloaddition with another diazo compound. Intriguingly, synthesis of substituted benzo[ de]chromenes and their decarboxylation products could be realized by controlling the reaction conditions. These reactions have a broad range of substrates, moderate to good yields, and high regioselectivity.


The Medication Optimization Value Proposition: Aligning Teams and Education to Improve Care.
United States health care lags behind other countries in quality and cost. The present health care system is unsustainable, and there is now a quick movement toward value-based care. This article lays out essential care delivery elements, and makes the case for medication optimization to enable new value-based models. Success factors include enhancing team-based care and interdisciplinary education to achieve patient-centered care.


Crystal structure of a designed, thermostable, heterotrimeric coiled coil.
Electrostatic interactions are often critical for determining the specificity of protein-protein complexes. To study the role of electrostatic interactions for assembly of helical bundles, we previously designed a thermostable, heterotrimeric coiled coil, ABC, in which charged residues were employed to drive preferential association of three distinct, 34-residue helices. To investigate the basis for heterotrimer specificity, we have used multiwavelength anomalous diffraction (MAD) analysis to determine the 1.8 A resolution crystal structure of ABC. The structure shows that ABC forms a heterotrimeric coiled coil with the intended arrangement of parallel chains. Over half of the ion pairs engineered to restrict helix associations were apparent in the experimental electron density map. As seen in other trimeric coiled coils, ABC displays acute knobs-into-holes packing and a buried anion coordinated by core polar amino acids. These interactions validate the design strategy and illustrate how packing and polar contacts determine structural uniqueness.


Antigenic conversion from HBcAg to HBeAg by degradation of hepatitis B core particles.
Highly purified and homogenous hepatitis B core particles were obtained from an autopsy liver. The core particles consisted mainly of an electrophoretically single major polypeptide with a molecular weight of 20,000 daltons. When antigenic conversion from hepatitis B core antigen (HBcAg) to hepatitis B e antigen was achieved by treating these core particles by sonication or by passing them through an anti-HBc IgG-conjugated Sepharose 4B column, no appreciable changes were found in the above protein composition. The same antigenic conversion was also achieved by centrifugation of core particles in CsCl, which revealed the process of morphological disintegration accompanied by antigenic conversion. These observations may support the hypothesis that HBcAg resides on a protein conformation which consists of a polypeptide sharing HBe antigenicity.


A clinical study of the effectiveness of two different 10% carbamide peroxide bleaching products: a 6-month followup.
The purpose of this study was to evaluate the efficacy of two different 10% carbamide peroxide bleaching products just after treatment and after a 6-month follow-up period. Methods. Two 10% carbamide peroxide products (Opalescence PF and Nite White ACP) were applied nightly for 14 days, according to the manufacturers' instructions. The color of teeth 11 and 21 of thirty-four subjects having A2 or darker teeth were measured with a spectrophotometer (L(∗); a(∗); b(∗)) before treatment, just after treatment (14 days) and after 6 months. Results and Conclusions. Both products produced significant whitening of teeth with total color change (ΔE(ab) (∗)) of approximately 5.20 units. There was a significant improvement in all 3 color coordinates (L*, a*, and b(∗)) for up to 6 months postbleaching (P < .05). Nite White showed a higher degree of relapse (27%) than Opalescence (18%) over the 6-month period. It is suggested that rebleaching after 6 months is not necessary.


Relapse: a reappraisal of assessment of outcome in schizophrenia.
Although measurement of the course of schizophrenic disorder has relied heavily on the concept of relapse, a review of the literature reveals a paucity of clear definitions of relapse. In most research studies, the assessment of relapse has been made on the basis of clinical judgment, with little attempt to apply more rigorous psychometric standards. The limitations of this approach are discussed and several possible improvements are suggested. These include: operational definitions that specify the qualitative, quantitative, and temporal characteristics of symptomatic exacerbation; avoidance of potentially confounding social variables; and the use of standardized rating scales and target symptoms.


Evaluation of endothelial function and arterial stiffness in HIV-infected patients: a pilot study.
In the era of combination antiretroviral therapy (cART), life expectancy of HIV-infected patients is the same as that of the general population, resulting in increasing prevalence of cardiovascular disease in this patient group. To assess the prevalence of endothelial dysfunction in HIV-infected patients and to identify factors which affect endothelial function and arterial stiffness. Thirty-seven adult HIV-infected patients, regardless of the fact and the type of cART, were enrolled into the study. In patient, reactive hyperaemia peripheral arterial tonometry assessment was performed using the Endo-PAT2000 device (ITAMAR®). This method allows evaluation of endothelial function ant arterial stiffness. Final analysis included 37 patients (median age 38 years, range 32-45 years), including 89.2% men. Endothelial dysfunction was found in 13 (35.1%) HIV-infected patients. We found no differences in demographic and clinical characteristics, laboratory data, and cardiovascular drug therapy between patients with or without endothelial dysfunction, except for platelet count which was higher in patients with endothelial dysfunction (174 [119-193] × 10³/mm3 vs. 222 [168-266] × 10³/mm³, p = 0.03). No demographic or clinical variables were identified as predictors of endothelial dysfunction in HIV-infected patients. In addition, no association was found between factors related to HIV infection, chronic drug therapy and the risk of endothelial dysfunction. Statistically significant correlations were found between arterial stiffness and age (rs = 0.53, p < 0.001), red blood cell count (rs = -0.39, p = 0.018), and platelet count (rs = 0.42, p = 0.009). CD4+ and CD8+ lymphocyte count and viral load were similar in patients with or without endothelial dysfunction. Arterial stiffness was significantly higher in patients with higher viral load (rs = -0.39, p = 0.0018) and in those with established AIDS (9.5 [1.0-16.0] vs. -5 [-10-5], p = 0.009). cART had no effect on endothelial dysfunction, while arterial stiffness was higher in patients treated with cART (10 [0-15] vs. -5 [-10-3], p = 0.014). Endothelial dysfunction is common in HIV-infected patients. In general, none of the analysed factors had an effect on endothelial function but cART had a negative effect on arterial stiffness.


Quantitative analysis of regional endocardial geometry dynamics from 4D cardiac CT images: endocardial tracking based on the iterative closest point with an integrated scale estimation.
Regional cardiac function analysis is important for the diagnosis and treatment planning of ischemic heart disease, but has not been sufficiently developed in the field of computed tomography (CT). Therefore, we propose a 3D endocardial tracking framework for cardiac CT using local point cloud registration based on the iterative closest point with an integrated scale estimation algorithm. We also introduce regional function descriptors that express the curvature and stretching of the endocardium: Surface distortion (E) and Scaling rate (S). For a region-to-region comparison, we propose endocardial segmentation according to coronary perfusion territories defined by the Voronoi partition based on coronary distribution. Our study of 65 endocardial segments in ten subjects showed that global endocardial deformation has a positive relationship with the stroke volume index (r = 0.896 and 0.829 in [Formula: see text] and [Formula: see text], respectively) and ejection fraction (r = 0.804 and 0.835), and a positive relationship with the brain natriuretic peptide level (r = 0.690 and 0.776). A positive relationship between segmental E and S (r = 0.845), a higher value of E in ischemic segments (p = 0.021) that are determined by fractional flow reserve estimated from coronary CT data, and a higher value of S in the left circumflex artery territory (p < 0.05) were also observed. The required radiation dose was 5.0 ± 0.7 mSv and the computation time was 7.2 ± 1.1 min. The result suggests that proposed endocardial deformation analysis using CT can be conducted on site and in time for the acute setting, and may be useful for the diagnosis of cardiac dysfunction or myocardial ischemia.


Light-addressable electrodeposition of cell-encapsulated alginate hydrogels for a cellular microarray using a digital micromirror device.
This paper describes a light-addressable electrolytic system used to perform an electrodeposition of calcium alginate hydrogels using a digital micromirror device (DMD). In this system, a patterned light illumination is projected onto a photoconductive substrate serving as a photo-anode to electrolytically produce protons, which can lead to a decreased pH gradient. The low pH generated at the anode can locally release calcium ions from insoluble calcium carbonate (CaCO(3)) to cause gelation of calcium alginate through sol-gel transition. By controlling the illumination pattern on the DMD, a light-addressable electrodeposition of calcium alginate hydrogels with different shapes and sizes, as well as multiplexed micropatterning was performed. The effects of the concentration of the alginate and CaCO(3) solutions on the dimensional resolution of alginate hydrogel formation were experimentally examined. A 3 × 3 array of cell-encapsulated alginate hydrogels was also successfully demonstrated through light-addressable electrodeposition. Our proposed method provides a programmable method for the spatiotemporally controllable assembly of cell populations into cellular microarrays and could have a wide range of biological applications in cell-based biosensing, toxicology, and drug discovery.


Fate of patients with acute myocardial infarction with patency of the infarct-related vessel achieved with successful thrombolysis versus rescue angioplasty.
Patients with failure of infarct-related artery recanalization after thrombolytic therapy have a poor clinical outcome. These patients have been considered for rescue angioplasty 90 min after thrombolytic therapy at the time of emergency catheterization in the course of five Thrombolysis and Angioplasty in Myocardial Infarction (TAMI) trials. The outcome of 776 patients with patent infarct-related vessels after emergency catheterization was analyzed--607 with thrombolysis-mediated patency of the infarct-related vessel and 169 with patency achieved by angioplasty. Baseline characteristics of the thrombolysis and angioplasty patency groups were similar except for a higher acute left ventricular ejection fraction (51.3% versus 48.2%) in the thrombolysis group (p = 0.003). Seven to 10 day left ventricular ejection fraction was higher (52.3% versus 48.1%), infarct zone functional recovery was greater (0.44 versus 0.21 standard deviation/chord, or 18% versus 7%, p = 0.001) and reocclusion was less (11% versus 21%) in the thrombolysis compared with the angioplasty group. Despite these differences, angioplasty patency was associated with the same low in-hospital mortality rate (5.9% versus 4.6%) and long-term mortality rate (3% versus 2%) as thrombolysis patency. Reocclusion adversely affected the mortality rate and ventricular functional recovery. Technical failure of rescue angioplasty was associated with a much higher mortality rate than was technical success (39.1% versus 5.9%). Thrombolysis patency was preferable to angioplasty patency after thrombolytic therapy in acute myocardial infarction, but both were associated with the same low in-hospital and long-term mortality rates, suggesting that rescue angioplasty is beneficial in some patients with failure of infarct-related artery recanalization after thrombolytic therapy.


T4-locally advanced nasopharyngeal carcinoma: prognostic influence of cranial nerve involvement in different radiotherapy techniques.
Cranial nerve involvement at disease presentation of nasopharyngeal carcinoma was not uncommon. We investigated the prognosis of patients with T4-locally advanced NPC, with or without cranial nerve involvement, and compared the outcome of patients treated using different radiotherapy techniques. In this retrospective study, 83 T4-locally advanced NPC patients were diagnosed according to the seventh edition of the American Joint Committee on Cancer staging system. All patients were treated using three-dimensional conformal radiotherapy (3D-CRT) or intensity-modulated radiation therapy (IMRT). The survival rate was analyzed using the Kaplan-Meier method. The 5-year overall, locoregional-free, and disease-free survival rates of patients treated using IMRT were 88.9%, 75.2%, and 69.2%, respectively. The outcome in these patients was significantly better than that in patients treated using 3D-CRT, with survival rates of 58.2%, 54.4%, and 47.2%, respectively. There was no significant difference in the 5-year overall, locoregional-free, and disease-free survival rates of the patients with (64.2%, 60.5%, and 53.5%, resp.) and without (76.9%, 63.6%, and 57.6%, resp.) cranial nerve involvement. Locally advanced NPC patients treated using IMRT had significantly better outcomes than patients treated using 3D-CRT. Our results showed that the outcome of T4 NPC patients with or without cranial nerve involvement was not different.


Channel formation properties of synthetic pardaxin and analogues.
Six analogues of teh 33-residue shark repellent neurotoxin pardaxin were synthesized by the solid phase method: [Ala13]pardaxin, [Gly14,Gly15]pardaxin, des[1----9]pardaxin, [N1-succinamido]pardaxin, C33-dihydroxyethylamido]pardaxin, and C33-[diaminoethylamido]pardaxin. The spectroscopic and functional characterizations of the analogues are described. The peptides were characterized spectroscopically by circular dichroism (CD) before and after binding to soybean vesicles. They were characterized functionally by measuring their potential to evoke the dissipation of diffusion potential and calcein release from sonicated unilamellar soybean liposomes, by determining their ability to create single channels in planar bilayers, and by measuring their cytolytic activity on human erythrocytes. The behavior of the analogues modified at the C terminus is similar to that of pardaxin. [N'-succinamido]Pardaxin, however, reveals an increase in alpha-helicity both alone and in the presence of liposomes. It has the same potency as pardaxin to dissipate diffusion potential, to evoke calcein release and to produce single channels in lipid bilayers, but at a slower rate than that of pardaxin. It has more than 70-fold less cytolytic activity than pardaxin. [Ala13] Pardaxin has twice the alpha-helical content than pardaxin, both alone and in the presence of vesicles, yet it has less effect on the diffusion potential and calcein release, and it does not have cytolytic activity on human erythrocytes. Both [Gly14,Gly15]pardaxin and des[1----9]pardaxin are much less potent than pardaxin in all effects. However des[1----9]pardaxin exhibits a slight change in alpha-helicity upon binding to vesicles, whereas [Gly14,Gly15]pardaxin does not. The results support a model in which pardaxin is composed of two putative alpha-helices separated by proline. The N-terminal alpha-helix is important for the insertion of the peptide to the lipid bilayer, and the C-terminal amphiphilic alpha-helix is the ion channel lining segment of pardaxin.


Natural antibodies bridge innate and adaptive immunity.
Natural Abs, belonging to isotypes IgM, IgG3, and IgA, were discovered nearly half a century ago. Despite knowledge about the role of the polyreactive natural IgM in pathogen elimination, B cell survival and homeostasis, inflammatory diseases, and autoimmunity, there is a lack of clarity about the physiological role of natural IgG and natural IgA because they appear incapable of recognizing Ags on their own and are perceived as nonreactive. However, recent research revealed exciting functions of natural IgG in innate immunity. Natural IgG:lectin collaboration swiftly and effectively kills invading pathogens. These advances prompt further examination of natural Abs in immune defense and homeostasis, with the potential for developing novel therapeutics. This review provides new insights into the interaction between natural Abs and lectins, with implications on how interactions between molecules of the innate and adaptive immune systems bridge these two arms of immunity.


Reduced fitness and physical functioning are long-term sequelae after curative treatment for esophageal cancer: a matched control study.
Reduced physical functioning is common following resections for esophageal cancer; however, objective data on physical performance outcomes in this cohort are rare. The aim of this study was to assess the physical performance and health related quality of life (HRQOL) of disease free survivors and compare findings in a case matched noncancer control group. Twenty-five males (mean (±SD) aged 63 (±6) years) who were over 6 months postesophagectomy and disease-free were compared with 25 controls (60 ± 6 years). Physical functioning was assessed through hand grip strength (dynamometry), exercise capacity (incremental shuttle walk test), physical activity levels (RT3 accelerometer), and body composition (bio-electrical impedance analysis). Health-related quality of life was measured using the EORTC QLQ-C30 questionnaire. Esophageal cancer survivors demonstrated significantly lower fitness (P < 0.001) and time spent in moderate (P < 0.001) and vigorous (P < 0.001) intensity physical activity compared with controls. Global health status and quality of life were similar in both groups (P = 0.245); however, physical and role functioning domains were lower in the cancer survivors (P < 0.001, and P = 0.001, respectively). These data show that disease-free survivors of curative esophageal cancer treatment demonstrate a significant compromise in physical functioning compared with controls, thus highlighting the multiple, complex rehabilitative needs of this cohort.


Opioid Use Disorder Documented at Delivery Hospitalization - United States, 1999-2014.
Opioid use by pregnant women represents a significant public health concern given the association of opioid exposure and adverse maternal and neonatal outcomes, including preterm labor, stillbirth, neonatal abstinence syndrome, and maternal mortality (1,2). State-level actions are critical to curbing the opioid epidemic through programs and policies to reduce use of prescription opioids and illegal opioids including heroin and illicitly manufactured fentanyl, both of which contribute to the epidemic (3). Hospital discharge data from the 1999-2014 Healthcare Cost and Utilization Project (HCUP) were analyzed to describe U.S. national and state-specific trends in opioid use disorder documented at delivery hospitalization. Nationally, the prevalence of opioid use disorder more than quadrupled during 1999-2014 (from 1.5 per 1,000 delivery hospitalizations to 6.5; p<0.05). Increasing trends over time were observed in all 28 states with available data (p<0.05). In 2014, prevalence ranged from 0.7 in the District of Columbia (DC) to 48.6 in Vermont. Continued national, state, and provider efforts to prevent, monitor, and treat opioid use disorder among reproductive-aged and pregnant women are needed. Efforts might include improved access to data in Prescription Drug Monitoring Programs, increased substance abuse screening, use of medication-assisted therapy, and substance abuse treatment referrals.


Methylated eyes absent 4 (EYA4) gene promotor in non-neoplastic mucosa of ulcerative colitis patients with colorectal cancer: evidence for a field effect.
Aberrant methylation of the EYA4 gene (mEYA4) highly discriminates ulcerative colitis (UC) cases with colorectal neoplasia from UC controls in both tissue and stool. It is not known if mEYA4 is also present in nonadjacent non-neoplastic mucosa (NNM) of UC patients with colorectal neoplasia. Formalin-fixed tissues from 25 UC cases with colorectal cancer (CRC) and 25 UC controls with neither CRC nor dysplasia were matched on gender, age, disease duration, disease extent, and coexistence of primary sclerosing cholangitis. DNA was extracted from sections of CRC and NNM from cases and UC control mucosae. Bisulfite-treated DNA was amplified using real-time methylation-specific PCR. The Wilcoxon rank-sum test assessed differences in mEYA4 levels from CRC, NNM, and control samples. Logistic regression was used to estimate sensitivity and specificity. Sufficient DNA was available for 20 cases and 20 controls. The median mEYA4 level (with interquartile range) was 2 (0-5.7) in control mucosae but 93 (38.5-306) in CRC (P < 0.0001) and 27.4 (3-140) in NNM (P = 0.0009). At 95% specificity, mEYA4 was present in 80% of CRC and 50% of NNM tissue samples. The odds ratio of mEYA4 in NNM as an indicator of synchronous CRC was 19 (95% confidence interval, 2-170). The authors demonstrate significantly higher mEYA4 levels in NNM and synchronous CRC from UC cases than in colorectal mucosae of UC controls without neoplasia. The finding of this CRC-associated field change has important implications to the use of mEYA4 as a potential UC surveillance marker in tissue or stool.


Effect of uncontrolled diabetes on plasma ghrelin concentrations and ghrelin-induced feeding.
Plasma levels of the orexigenic hormone, ghrelin, decrease rapidly on nutrient ingestion and yet are paradoxically elevated in rats with hyperphagia induced by streptozotocin-induced diabetes (STZ-DM). In the current work, we investigated the mechanisms underlying the relationships among uncontrolled diabetes, food intake, and plasma ghrelin concentrations in an effort to clarify whether increased ghrelin signaling contributes to diabetic hyperphagia. Whereas food intake did not increase until d 3 after STZ administration, plasma ghrelin levels were increased by more than 2-fold (P < 0.05) on d 1. As hyperphagia developed, however, plasma ghrelin levels declined steadily. Because this reduction of plasma ghrelin levels was reversed by matching food intake of STZ-DM rats to that of nondiabetic controls, our results demonstrated that the effect of uncontrolled diabetes to increase plasma ghrelin levels is partially offset by hyperphagic feeding. In addition, we found that although intragastric nutrient infusion rapidly and comparably decreased plasma ghrelin levels in both groups (by 46-49%; P < 0.05), this effect was short lived in STZ-DM rats relative to nondiabetic controls (60 min vs. 120 min; P < 0.05). We further demonstrated that in rats with STZ-DM, food intake increased by 357% (P < 0.05) in response to intracerebroventricular administration of ghrelin at a dose that was subthreshold for feeding effects in nondiabetic controls. Collectively, these findings demonstrate that uncontrolled diabetes increases both circulating ghrelin levels and behavioral sensitivity to ghrelin. Although plasma ghrelin levels fall in response to hyperphagic feeding, these findings support the hypothesis that increased ghrelin signaling contributes to the pathogenesis of diabetic hyperphagia.


Are calcium-dependent protein kinases involved in the regulation of glycolytic/gluconeogenetic enzymes? Studies with Ca2+/calmodulin-dependent protein kinase and protein kinase C.
Changes in glycolytic flux have been observed in liver under conditions where effects of cAMP seem unlikely. We have, therefore, studied the phosphorylation of four enzymes involved in the regulation of glycolysis and gluconeogenesis (6-phosphofructo-1-kinase from rat liver and rabbit muscle; pyruvate kinase, 6-phosphofructo-2-kinase and fructose-1,6-bisphosphatase from rat liver) by defined concentrations of two cAMP-independent protein kinases: Ca2+/calmodulin-dependent protein kinase and Ca2+/phospholipid-dependent protein kinase (protein kinase C). The results were compared with those obtained with the catalytic subunit of cAMP-dependent protein kinase. The following results were obtained. 1. Ca2+/calmodulin-dependent protein kinase phosphorylates 6-phosphofructo-1-kinase and L-type pyruvate kinase at a slightly lower rate as compared to cAMP-dependent protein kinase. 2. 6-Phosphofructo-1-kinase is phosphorylated by the two kinases at a single identical position. There is no additive phosphorylation. The final stoichiometry is 2 mol phosphate/mol tetramer. The same holds for L-type pyruvate kinase except that the stoichiometry with either kinase or both kinases together is 4 mol phosphate/mol tetramer. 3. Rabbit muscle 6-phosphofructo-1-kinase is phosphorylated by cAMP-dependent protein kinase but not by Ca2+/calmodulin-dependent protein kinase. 4. Fructose-1,6-bisphosphatase from rat but not from rabbit liver is phosphorylated at the same position but at a markedly lower rate by Ca2+/calmodulin-dependent protein kinase when compared to the phosphorylation by cAMP-dependent protein kinase. 5. 6-Phosphofructo-2-kinase is phosphorylated by Ca2+/calmodulin-dependent protein kinase only at a negligible rate. 6. Protein kinase C does not seem to be involved in the regulation of the enzymes examined: only 6-phosphofructo-2-kinase became phosphorylated to a significant degree. In contrast to the phosphorylation by cAMP-dependent protein kinase, this phosphorylation is not associated with a change of enzyme activity. This agrees with our observation that the sites of phosphorylation by the two kinases are different. The results indicate that Ca2+/calmodulin-dependent protein kinase but not protein kinase C could be involved in the regulation of hepatic glycolytic flux under conditions where changes in the activity of cAMP-dependent protein kinase seem unlikely.


Identification of human papillomavirus DNA gene sequences in human breast cancer.
Human papilloma viruses (HPVs) are accepted as being carcinogenic in human cervical and anogenital cancers. The suspicion that HPVs may also have a role in human breast cancer is based on the identification of HPVs in human breast tumours and the immortalisation of normal human breast cells by HPV types 16 and 18. For this investigation, DNA that had been previously extracted and fresh frozen at -70 degrees C from 50 unselected invasive ductal breast cancer specimens were screened by polymerase chain reaction (PCR) for HPV type 16, 18 and 33 gene sequences. We show that HPV 18 gene sequences are present in DNA extracted from breast tumours in Australian women. Overall, 24 (48%) of the 50 samples were HPV positive. Overall no correlations with tumour grade, patient survival, steroid receptor status, ERB-2, p53 expression and mutation were observed. Human papilloma viruses may have a role in human breast cancer. We speculate that HPVs may be transmitted by hand from the female perineum to the breast.


LRRK2-Related Parkinson's Disease Due to Altered Endolysosomal Biology With Variable Lewy Body Pathology: A Hypothesis.
Mutations in the gene encoding for leucine-rich repeat kinase 2 (LRRK2) are associated with both familial and sporadic Parkinson's disease (PD). LRRK2 encodes a large protein comprised of a GTPase and a kinase domain. All pathogenic variants converge on enhancing LRRK2 kinase substrate phosphorylation, and distinct LRRK2 kinase inhibitors are currently in various stages of clinical trials. Although the precise pathophysiological functions of LRRK2 remain largely unknown, PD-associated mutants have been shown to alter various intracellular vesicular trafficking pathways, especially those related to endolysosomal protein degradation events. In addition, biochemical studies have identified a subset of Rab proteins, small GTPases required for all vesicular trafficking steps, as substrate proteins for the LRRK2 kinase activity in vitro and in vivo. Therefore, it is crucial to evaluate the impact of such phosphorylation on neurodegenerative mechanisms underlying LRRK2-related PD, especially with respect to deregulated Rab-mediated endolysosomal membrane trafficking and protein degradation events. Surprisingly, a significant proportion of PD patients due to LRRK2 mutations display neuronal cell loss in the substantia nigra pars compacta in the absence of any apparent α-synuclein-containing Lewy body neuropathology. These findings suggest that endolysosomal alterations mediated by pathogenic LRRK2 per se are not sufficient to cause α-synuclein aggregation. Here, we will review current knowledge about the link between pathogenic LRRK2, Rab protein phosphorylation and endolysosomal trafficking alterations, and we will propose a testable working model whereby LRRK2-related PD may present with variable LB pathology.


Non-linear dependence of the dielectric properties of chick embryo myoblast membranes exposed to a sinusoidal 50 Hz magnetic field.
Primary chick embryo myoblasts can be a useful tool for studying the developmental events which accompany myoblast differentiation, particularly myoblast membrane fusion. To determine whether the electrical properties and/or fusion in these systems are affected by 50 Hz magnetic fields, chick embryo myoblast cultures were exposed to B-field intensities ranging from 1 to 10 mT. The electrical parameters of the myoblasts, i.e. membrane conductivity, membrane permittivity and the conductivity of the cell interior (cytosol) were determined by the analysis of conductivity dispersion data in the radio frequency range (10 kHz-100 MHz). Preliminary results indicate that the time of fusion (60 h) is not affected by these fields, but that the absolute values of the two membrane electrical parameters are affected. In particular, a B-field intensity-dependent decrease was observed. The maximum effect resulted after a 1 h exposure to a magnetic flux density of about 5 mT. The conductivity of the cytosol remained unchanged. These data seem to indicate that exposure to 50 Hz magnetic fields affects both static and dynamic membrane properties in primary chick embryo myoblasts.


In silico design of novel probes for the atypical opioid receptor MRGPRX2.
The primate-exclusive MRGPRX2 G protein-coupled receptor (GPCR) has been suggested to modulate pain and itch. Despite putative peptide and small-molecule MRGPRX2 agonists, selective nanomolar-potency probes have not yet been reported. To identify a MRGPRX2 probe, we first screened 5,695 small molecules and found that many opioid compounds activated MRGPRX2, including (-)- and (+)-morphine, hydrocodone, sinomenine, dextromethorphan, and the prodynorphin-derived peptides dynorphin A, dynorphin B, and α- and β-neoendorphin. We used these to select for mutagenesis-validated homology models and docked almost 4 million small molecules. From this docking, we predicted ZINC-3573-a potent MRGPRX2-selective agonist, showing little activity against 315 other GPCRs and 97 representative kinases-along with an essentially inactive enantiomer. ZINC-3573 activates endogenous MRGPRX2 in a human mast cell line, inducing degranulation and calcium release. MRGPRX2 is a unique atypical opioid-like receptor important for modulating mast cell degranulation, which can now be specifically modulated with ZINC-3573.


A Toxin-Antitoxin System VapBC15 from Synechocystis sp. PCC 6803 Shows Distinct Regulatory Features.
Type II toxin-antitoxin (TA) systems play important roles in bacterial stress survival by regulating cell growth or death. They are highly abundant in cyanobacteria yet remain poorly characterized. Here, we report the identification and regulation of a putative type II TA system from Synechocystis PCC6803, VapBC15. The VapBC15 system is encoded by the chromosomal operon vapBC15. Exogenous expression of VapC15 dramatically arrested cell growth of Escherichia coli and reduced the numbers of colony-forming units (CFU). The VapC15 toxicity could be which was counteracted neutralized by simultaneous or delayed production of VapB15. Biochemical analysis demonstrated the formation of VapB15-VapC15 complexes by the physical interaction between VapB15 and VapC15. Notably, the VapB15 antitoxin up-regulated the transcription of the vapBC15 operon by directly binding to the promoter region, and the VapC15 toxin abolished the up-regulatory effect by destabilizing the binding. Moreover, VapB15 can be degraded by the proteases Lons and ClpXP2s from Synechocystis PCC6803, thus activating the latent toxicity of VapBC15. These findings suggest that VapBC15 represents a genuine TA system that utilizes a distinct mechanism to regulate toxin activity.


Mutation spectrum of the OPA1 gene in a large cohort of patients with suspected dominant optic atrophy: Identification and classification of 48 novel variants.
Autosomal dominant optic atrophy is one of the most common inherited optic neuropathies. This disease is genetically heterogeneous, but most cases are due to pathogenic variants in the OPA1 gene: depending on the population studied, 32-90% of cases harbor pathogenic variants in this gene. The aim of this study was to provide a comprehensive overview of the entire spectrum of likely pathogenic variants in the OPA1 gene in a large cohort of patients. Over a period of 20 years, 755 unrelated probands with a diagnosis of bilateral optic atrophy were referred to our laboratory for molecular genetic investigation. Genetic testing of the OPA1 gene was initially performed by a combined analysis using either single-strand conformation polymorphism or denaturing high performance liquid chromatography followed by Sanger sequencing to validate aberrant bands or melting profiles. The presence of copy number variations was assessed using multiplex ligation-dependent probe amplification. Since 2012, genetic testing was based on next-generation sequencing platforms. Genetic screening of the OPA1 gene revealed putatively pathogenic variants in 278 unrelated probands which represent 36.8% of the entire cohort. A total of 156 unique variants were identified, 78% of which can be considered null alleles. Variant c.2708_2711del/p.(V903Gfs*3) was found to constitute 14% of all disease-causing alleles. Special emphasis was placed on the validation of splice variants either by analyzing cDNA derived from patients´ blood samples or by heterologous splice assays using minigenes. Splicing analysis revealed different aberrant splicing events, including exon skipping, activation of exonic or intronic cryptic splice sites, and the inclusion of pseudoexons. Forty-eight variants that we identified were novel. Nine of them were classified as pathogenic, 34 as likely pathogenic and five as variant of uncertain significance. Our study adds a significant number of novel variants to the mutation spectrum of the OPA1 gene and will thereby facilitate genetic diagnostics of patients with suspected dominant optic atrophy.


The wicked problem of B(A)ME degree award gaps and systemic racism in our universities.
The independent regulator for higher education in England, the Office for Students (OfS), set new national targets in late 2018 to achieve equality of opportunity in higher education by tackling degree award gaps. The sector response to this was to measure degree award gaps between B(A)ME students and white students in their higher education institutions. Analysis of degree award gaps using quantitative methods has revealed an "unexplained gap". We argue that the existence of this "unexplained gap" is evidence of "systemic racism". However, the factors influencing a degree award and their associated gaps across different racialized groups of students are so complex, that its problematisation, never-mind its solution is inherently complex. It is our view, therefore, racialized degree award gaps are a wicked problem. Despite this, it is also our view that it is an important social justice endeavor that we must still seek to address as a sector. To do so, we propose a mixed methods approach that uses dynamic centring and an intersectional lens to better understand the experiences of racialized students within the higher education "system". Current quantitative analysis of degree award gaps simply tells us how different groups of racialized students experience the system. In using a mixed methods approach in the way we outline, we may better understand the racialized lived experience of our students and the factors influencing the experience of different racialized groups within the "system". This solution-focused approach can help create opportunities that enable students to better navigate social structures and systems and improve their experience in the system. However, this will not address the wicked problem of degree award gaps itself, which is complex, pervasive, and messy.


High throughput encapsulation of murine fibroblasts in alginate using the JetCutter technology.
The JetCutter technology originally developed for high-throughput encapsulation of particles and substances into small beads was applied in this study for the entrapment of mammalian cells in alginate beads. In contrast to other established techniques such as the air jet droplet generation or laminar jet break-up, the JetCutter is capable of working with highly viscous fluids necessary for the production of stable beads based on hydrogels. A 1.5% (w/v) sodium alginate solution containing 2.0 x 106 murine fibroblasts/ml was processed under good manufacturing practice (GMP) conditions to beads with a mean diameter of 320 microm. The production capacity of the JetCutter technology was 5200 beads/s or to approximately 330 ml bead suspension per h. Beads were coated with poly-L-lysine and with an additional alginate layer to produce hollow microcapsules containing living cells. The influence of this method of encapsulation on the cell viability and morphology was investigated by light microscopic techniques. Encapsulated cells showed unchanged rates of proliferation and preserved morphology. They were able to survive in culture for extended periods of time. In conclusion, the JetCutter technology seems to be well suitable for alginate bead encapsulation of living mammalian cells.


[Combined and extended radical operations in colorectal cancer patients].
This is a report on radical operative interventions performed in 513/755 patients presenting complicated colorectal carcinoma, covering the period 2000 through 2007. One hundred and four patients are subjected to combined and extended operations, distributed as follows: 61 combined and 43 extended. In thirty three patients with abdominoperineal extirpation the combined interventions include: hysterectomy (7), ovariectomy (9), resection of vagina (8), prostate gland resection (5) and bladder resection (4). In case of resection of rectum after Hartmann, combined intervention is done in seventeen patients - small intestine resection - 7, ovariectomy - 2, appendectomy - 3 and cyst extirpation from lethality amounts to 12.9 per cent of patients with complicated colorectal carcinoma undergoing combined and extended surgical interventions.


Pharmacotherapy of depression in old age.
This review addresses the pharmacotherapy of antidepressants in the elderly. We performed a search based on Medline and the Cochrane Library. In addition to a meta-analysis of 17 randomised controlled studies, 36 randomised controlled trials of patients over the age of 60 published between 1980 and 2005 in English met the selection criteria and were included. Existing evidence suggests that no one class of antidepressant drugs has been found to be more effective than another in the treatment of depression in the elderly. Although newer antidepressants are not more effective than older ones, they are better tolerated and are safe especially in overdose. The adverse effect data suggest modest superiority of selective serotonin reuptake inhibitors over tricyclic antidepressants. The evidence available indicates that antidepressant treatment of four weeks has a beneficial effect compared to placebo. As to prevention of relapse and recurrence, antidepressants should be continued for at least six months after good initial response. In patients with high risk of relapse, treatment should be continued for at least two years. Long-term efficacy has been shown for dosulepin, nortriptyline and citalopram. In patients with dementia with persistent and significant symptoms antidepressant treatment may be indicated. At present, clomipramine, citalopram and sertraline have been reported as being superior to placebo. There is a paucity of data on the use of antidepressants in very elderly individuals, patients with significant comorbidity and patients with dementia. More data on the effect of antidepressants in the elderly, especially in the over 80-age group are needed.


Renner-Teller interactions in the vibrational autoionization of polyatomic molecules.
Vibrational autoionization induced by the Renner-Teller interaction in linear polyatomic molecules is considered in the context of the three-state electrostatic model developed by Gauyacq and Jungen [Mol. Phys. 41, 383 (1980)]. For small interactions, simple formulas are derived for the quantum defect matrix elements and the autoionization rates in terms of the more common Renner-Teller parameters derived from spectroscopic analyses of low-lying Rydberg states. These formulas should provide guidance for empirical fitting of quantum defect parameters to spectra of high Rydberg states. Consideration of typical values of the Renner-Teller parameters also allows the estimation of vibrational autoionization rates induced by these interactions. These estimates support the validity of the Deltav=-1 propensity rule for vibrational autoionization. Constraints on the vibrational autoionization rates for the symmetric stretching vibration are also discussed. In the following paper, electron capture by polyatomic molecular ions into vibrationally autoionizing Rydberg states is considered from the same perspective, and a simple formula is derived to allow the estimation of the effect of this process on dissociative recombination cross sections.


Reliability of laterality effects in a dichotic listening task with nonverbal material.
The present study examined the reliability of a dichotic listening task using nonverbal stimuli. Twenty undergraduate students (all right-handed native English speakers) had to report whether they had heard a target emotion. The task used English words (bower, dower, power, tower) pronounced with an angry, happy, neutral, or sad emotional tone. Results showed a relatively high level of test-retest reliability for the laterality effect. In addition, a significant gender by ear of presentation interaction was obtained. The interaction reflected the fact that a strong left ear advantage was found in females but not in males. The findings indicate that the task used here should be considered a reliable means to assess the lateralization of emotions. Issues concerning the relation between gender and laterality are addressed in the discussion.


Deciphering impedance cytometry signals with neural networks.
Microfluidic impedance cytometry is a label-free technique for high-throughput single-cell analysis. Multi-frequency impedance measurements provide data that allows full characterisation of cells, linking electrical phenotype to individual biophysical properties. To efficiently extract the information embedded in the electrical signals, potentially in real-time, tailored signal processing is needed. Artificial intelligence approaches provide a promising new direction. Here we demonstrate the ability of neural networks to decipher impedance cytometry signals in two challenging scenarios: (i) to determine the intrinsic dielectric properties of single cells directly from raw impedance data streams, (ii) to capture single-cell signals that are hidden in the measured signals of coincident cells. The accuracy of the results and the high processing speed (fractions of ms per cell) demonstrate that neural networks can have an important role in impedance-based single-cell analysis.


Unlocking the potential of microalgae cultivated on wastewater combined with salinity stress to improve biodiesel production.
Microalgae have the potential as a source of biofuels due to their high biomass productivity and ability to grow in a wide range of conditions, including wastewater. This study investigated cultivating two microalgae species, Oocystis pusilla and Chlorococcus infusionum, in wastewater for biodiesel production. Compared to Kühl medium, KC medium resulted in a significant fold increase in cellular dry weight production for both O. pusilla and C. infusionum, with an increase of 1.66 and 1.39, respectively. A concentration of 100% wastewater resulted in the highest growth for O. pusilla, with an increase in biomass and lipid content compared to the KC medium. C. infusionum could not survive in these conditions. For further increase in biomass and lipid yield of O. pusilla, different total dissolved solids (TDS) levels were used. Maximum biomass and lipid productivities were achieved at 3000 ppm TDS, resulting in a 28% increase in biomass (2.50 g/L) and a 158% increase in lipid yield (536.88 mg/g) compared to KC medium. The fatty acid profile of O. pusilla cultivated on aerated wastewater at 3000 ppm TDS showed a high proportion of desirable saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA) for biodiesel production. Cultivating microalgae in wastewater for biodiesel production can be cost-effective, especially for microalgae adapted to harsh conditions. It could be concluded that O. pusilla is a promising candidate for biodiesel production using wastewater as a growth medium, as it has high biomass productivity and lipid yield, and its fatty acid profile meets the standard values of American and European biodiesel standards. This approach offers a sustainable and environmentally friendly solution for producing biofuels while reducing the environmental impact of wastewater disposal.


Characterization of the Su antigen, a macromolecular complex of 100/102 and 200-kDa proteins recognized by autoantibodies in systemic rheumatic diseases.
The Su autoantigen was characterized biochemically using human and murine autoimmune sera and the clinical significance of anti-Su antibodies was studied in 236 Japanese and 160 American patients with systemic rheumatic diseases. Anti-Su in immunodiffusion (ID) was strongly associated with immunoprecipitation of one or more 100- to 102-kDa proteins by MRL/lpr mouse sera (27/32 of ID positive vs 4/20 of ID negative, P = 0.000016), and all four human anti-Su reference sera immunoprecipitated the 100/102-kDa protein(s). In addition, all sera immunoprecipitated a less efficiently labeled approximately 200-kDa protein that comigrated on sucrose density gradients with the 100/102-kDa proteins. Based on these data, a complex of the 100/102-kDa and 200-kDa proteins is likely to be the main target of anti-Su antibodies. Three of four anti-Su monospecific sera were negative for immunofluorescent antinuclear antibodies (ANA), suggesting anti-Su antibodies may be associated with a negative ANA in some cases. Autoantibodies to Su were detected frequently by immunoprecipitation in systemic lupus erythematosus (17-21%), scleroderma (13-20%), and overlap syndrome (22-40%) and were associated with autoantibodies to Ku.


Ageing and COVID-19: What is the Role for Elderly People?
Italy is one of the oldest countries in Europe and in the world and now it is also one of the first countries that are fighting against COVID-19. In our country, the increasing life expectancy (80.5 for males and 84.9 for females, with a total life expectancy of 82.9) has led to very positive consequences for health and the well-being of elderly people: a very high number of older adults lives and acts independently in their daily life, even if they have one or more than one chronic disease. In the time of COVID-19's outbreak in Italy, the focus of the media was on elderly people for two main reasons. First, many older people demonstrated a very high civic sense and they were helping society to fight against the pandemic. Second, also in Italy, like in China, the older adults are at higher risk in being infected with COVID-19 and if they get ill, they have a higher risk of death. The balance previously achieved between age-related disorders and a good quality of life and good health is now under high pressure. It is very important to protect elderly people from infection, but also it is important to respect them and to support them in this complex situation. There is a great risk of "ageism". In agreement with Lloyd-Sherlock and colleagues (2020), in this editorial we propose some hints of analysis, starting from the ongoing experience in Italy.


Evaluation of Root Canal Anatomy of Maxillary Premolars Using Swept-Source Optical Coherence Tomography in Comparison with Dental Operating Microscope and Cone Beam Computed Tomography.
This study aimed to evaluate the ability of swept-source optical coherence tomography (OCT) to detect internal anatomy of maxillary premolars in comparison with dental operating microscope (DOM) and cone beam computed tomography (CBCT). The ability of OCT to observe the pulp horn during cavity preparation and assess the remaining dentin thickness (RDT) has been demonstrated, whereas validation of OCT in comparison with other imaging techniques seems required. Ten extracted human maxillary premolars were sectioned perpendicular to the tooth axis from the occlusal surface at approximately 2 mm increments. OCT and DOM were performed after each cut, and microfocus X-ray computed tomography (micro-CT; reference standard) and CBCT were conducted before sectioning and after the first and second cuts. Three examiners evaluated all images for presence of the pulp horn/pulp chamber, isthmus, lateral canals, and the number of root canals. RDT was determined from OCT, micro-CT, and CBCT images. Correlations were analyzed with Pearson's correlation coefficient. OCT had a sensitivity and specificity of 0.90 and 0.80 in detecting the pulp horn/pulp chamber and 0.84 and 0.71 in detecting the isthmus, respectively. The three techniques showed strong correlations in detecting the number of root canals compared with micro-CT. OCT and DOM did not detect lateral canals. For RDT values, strong correlations were observed between micro-CT and CBCT, micro-CT and OCT, and CBCT and OCT (p < 0.01 for all). Under the present experimental condition, OCT accurately measured RDT and detected internal tooth anatomy such as the pulp horn, isthmus, and root canals.


Early Prediction of Mortality after Birth Asphyxia with the nSOFA.
(1) Birth asphyxia is a major cause of delivery room resuscitation. Subsequent organ failure and hypoxic-ischemic encephalopathy (HIE) account for 25% of all early postnatal deaths. The neonatal sequential organ failure assessment (nSOFA) considers platelet count and respiratory and cardiovascular dysfunction in neonates with sepsis. To evaluate whether nSOFA is also a useful predictor for in-hospital mortality in neonates (≥36 + 0 weeks of gestation (GA)) following asphyxia with HIE and therapeutic hypothermia (TH), (2) nSOFA was documented at ≤6 h of life. (3) A total of 65 infants fulfilled inclusion criteria for TH. All but one infant received cardiopulmonary resuscitation and/or respiratory support at birth. nSOFA was lower in survivors (median 0 [IQR 0-2]; n = 56, median GA 39 + 3, female n = 28 (50%)) than in non-survivors (median 10 [4-12], p < 0.001; n = 9, median GA 38 + 6, n = 4 (44.4%)). This was also observed for the respiratory (p < 0.001), cardiovascular (p < 0.001), and hematologic sub-scores (p = 0.003). The odds ratio for mortality was 1.6 [95% CI = 1.2-2.1] per one-point increase in nSOFA. The optimal cut-off value of nSOFA to predict mortality was 3.5 (sensitivity 100.0%, specificity 83.9%). (4) Since early accurate prognosis following asphyxia with HIE and TH is essential to guide decision making, nSOFA (≤6 h of life) offers the possibility of identifying infants at risk of mortality.


Battlefield advanced trauma life support (BATLS). Chapter 7. Abdominal injuries.
Recognise the possibility of abdominal injury. Two major types of abdominal injury may be present in war, blunt and penetrating. In either case, early surgery is essential. With blunt injuries, a high index of suspicion is required. Diagnosis in the early stages may be very difficult. Serial examination is required; even then, evacuate early. Aids to diagnosis include: DPL, FAST, CT Casualties with penetrating injuries must be given the highest priority for evacuation and surgical evaluation. Penetrating injury to the loin, back, buttocks, lower chest and thighs, should raise suspicion of intra-abdominal injury. Management of abdominal injury follows normal BATLS protocol and includes: Primary survey: Remember the A B C D E routine. Resuscitation: This involves: Continued oxygenation Appropriate therapy for hypovolaemic shock (refer to Chapter 5) Continued monitoring of response by assessment of vital signs, aided by additional parameters such as pulse oxymetry and blood gas analysis, when available. Placement of nasogastric tube and bladder catheter. Secondary survey: Following stabilization, but not before, you may proceed to full assessment of the mechanism and location of injury. This demands a full secondary survey and continued monitoring of the casualty.


[Cell phones: health risks and prevention].
The paper describes first of all the electromagnetic radiation of cellular phones and presents the physical parameters used to measure and evaluate the absorption of emissions of radio stations and cellular phones. It then presents selected research results of the experimental studies in vivo and in vitro which examine the biological effects of the emissions of cellular phones. The review of the epidemiologic evidence focuses in particular the epidemiologic studies on the use of cell phones and brain tumours, identifying some of the reasons of the conflicting results obtained. Studies dealing with the health risks involved in the increasing use of cellular phones by adolescents and children, more sensitive to this exposure, are also presented showing the need for special caution. The problem of hypersensitivity observed in some individuals is also briefly discussed. Finally the paper presents a summary of the main prevention measures necessary in order to reduce the risks in the framework of the "precautionary principle" including prevention policies and exposure limits in various countries.


Collapse in the Heat - From Overhydration to the Emergency Room - Three Cases of Exercise-Associated Hyponatremia Associated with Exertional Heat Illness.
Exertional heat illness and exercise-associated hyponatremia continue to be a problem in military and recreational events. Symptoms of hyponatremia can be mistaken for heat exhaustion or heat stroke. We describe three cases of symptomatic hyponatremia initially contributed to heat illnesses. The first soldier was a 31-yr-old female who "took a knee" at mile 6 of a 12-mile foot march. She had a core temperature of 100.9°F, a serum sodium level of 129 mmol/L, and drank approximately 4.5 quarts of water in 2 h. The second case was a 27-yr-old female soldier who collapsed at mile 11 of a 12-mile march. Her core temperature was 102.9°F and sodium level was 131 mmol/L. She drank 5 quarts in 2.5 h. The third soldier was a 27-yr-old male who developed nausea and vomiting while conducting an outdoor training event. His core temperature was 98.7°F and sodium level was 125 mmol/L. He drank 6 quarts in 2 h to combat symptoms of heat. All the three cases developed symptomatic hyponatremia by overconsumption of fluids during events lasting less than 3 h. Obtaining point-of-care serum sodium may improve recognition of hyponatremia and guide management for the patient with suspected heat illness and hyponatremia. Depending on severity of symptoms, exercise-associated hyponatremia can be managed by fluid restriction, oral hypertonic broth, or with intravenous 3% saline. Utilizing an ad libitum approach or limiting fluid availability during field or recreational events of up to 3 h may prevent symptomatic hyponatremia while limiting significant dehydration.


Phosphatase and Tensin Homolog (PTEN) Expression as a Surrogate Biomarker Correlated With the Depth of Invasion in Cutaneous Malignant Melanoma.
Objective The aim of this study is to evaluate the expression of the phosphatase and tensin homolog (PTEN), which is a tumor suppressor gene that is implicated in the pathogenesis of cutaneous malignant melanoma, in normal skin and melanoma tissue samples. The study also aimed to correlate PTEN expression levels with various clinicopathological parameters of melanoma lesions, thus highlighting the utility of PTEN expression as a prognostic biomarker for melanoma. Study design Immunohistochemistry (IHC) staining was performed on tissue microarray samples representing normal skin and melanoma biopsies of different clinicopathological parameters. Tissue photomicrographs were evaluated with Aperio ImageScope, which has a positive-pixel-counting algorithm built in. Subsequently, a histochemical score (H-score) was derived from the percentage of positive cells (%-staining) and their staining intensity. The H-scores were averaged in groups of tissue samples representing the different melanomas' tumor (T), node (N), and distant metastasis (M), also known as TNM parameters, as set forth by the American Joint Committee on Cancer (AJCC) classification. The mean H-scores were statistically compared using a two-tailed unpaired t-test. Results The PTEN protein expression was measured by IHC and found to be correlated with tumor thickness (T), which is a reliable indicator for survival rates. Specifically, PTEN was significantly downregulated in tumors with a thickness over 2 mm (T3+T4) compared to tumors with a thickness at or below 2 mm (T1+T2). Conclusions The PTEN protein expression, as measured by immunohistochemistry, helped differentiate between tumors with a thickness over 2 mm and tumors with a thickness at or below 2 mm, suggesting PTEN as a potential surrogate marker for the melanoma's invasion depth along with possible prognostic implications. Longitudinal studies evaluating risk stratification based on the expression of PTEN are needed to establish the utility of this promising biomarker in the clinic as an adjunct for pathological examination.


Theoretical Prediction of Robust Second-Row Oxyanion Clusters in the Metastable Domain of Antielectrostatic Hydrogen Bonding.
We provide ab initio and density functional theory evidence for a family of surprisingly robust like-charged clusters of common HSO4- and H2PO4- oxyanions, ranging up to tetramers of net charge 4-. Our results support other recent theoretical and experimental evidence for "antielectrostatic" hydrogen-bonded (AEHB) species that challenge conventional electrostatic conceptions and force-field modeling of closed-shell ion interactions. We provide structural and energetic descriptors of the predicted kinetic well-depths (in the range 3-10 kcal/mol) and barrier widths (in the range 2-4 Å) for simple AEHB dimers, including evidence of extremely strong hydrogen bonding in the fluoride-bisulfate dianion. For more complex polyanionic species, we employ natural-bond-orbital-based descriptors to characterize the electronic features of the cooperative hydrogen-bonding network that are able to successfully defy Coulomb explosion. The computational results suggest a variety of kinetically stable AEHB species that may be suitable for experimental detection as long-lived gas-phase species or structural units of condensed phases, despite the imposing electrostatic barriers that oppose their formation under ambient conditions.


Development and evaluation of immunochromatographic rapid tests for screening of cannabinoids, cocaine, and opiates in urine.
The test principle and the optimization of the reactive ingredients are described for the one-step dip and-read immunochromatographic FRONTLINE rapid tests for drugs-of-abuse testing in urine samples. In a multicenter evaluation the rapid tests were compared with FPIA and EMIT immunoassays. Discrepant results were further analyzed by gas chromatography-mass spectrometry methods. In the comparison of the cannabinoids rapid tests versus both immunoassays using clinical and forensic urine samples (399 versus FPIA and 755 versus EMIT), sensitivities and specificities were 97% or better for both comparisons. For cocaine, a sensitivity of 100% versus both routine technologies was obtained, whereas the specificity was reduced somewhat to 91% because of some cross-reactivity with metabolites of methadone and of clozapine. Specificity was very high for the cocaine rapid tests (98-100%) when applied to urine samples of persons not in a methadone maintenance program. Sensitivities and specificities for the opiates rapid tests were 99% or better at all sites when compared with the routine methods. In the screening of about 1200 clinical urine samples for cannabinoids, cocaine or opiates misuse only six samples would have stayed undetected by rapid test analyzes. These results show the FRONTLINE assays allow a reliable and immediate screening for drugs of abuse.


Anti-N-Methyl-D-Aspartic Acid Receptor 2 (Anti-NR2) Antibody in Neuropsychiatric Lupus Serum Damages the Blood-Brain Barrier and Enters the Brain.
BACKGROUND Brain microvessel endothelial cells constitute an important component in the blood-brain barrier. Cell-culture-based models of the blood-brain barrier (BBB) have been extensively applied in pharmacology, pathology and physiology. This study investigated effects of anti-N-methyl-D-aspartic acid receptor 2 (anti-NR2), N-methyl-D-aspartic acid (NMDA) receptor antibodies, NMDA receptor antagonists, and NMDA receptor agonists on brain microvessel endothelial cell models, and verified the effect of anti-NR2 antibody on the BBB as a receptor agonist. MATERIAL AND METHODS The primary brain microvessel endothelial cells were isolated and cultured, and an in vitro BBB model was established based on microvessel endothelial cells. Anti-NR2 antibody, glutamic acid, ifenprodil, and memantine were added in the BBB model to analyze changes in transepithelial electrical resistance (TEER) and to examine the permeability of the brain microvessel endothelial cell model. RESULTS The results showed that TEER values were significantly decreased by the addition of anti-NR2 antibody and glutamate, but were significantly increased by the addition of ifenprodil and memantine. TEER values showed no changes when treated by anti-NR2 antibody and ifenprodil, as well as anti-NR2 antibody and memantine. When dexamethasone was added, the TEER values increased by 23.8%, 39.4%, and 29.6% by treating with anti-NR2 antibody, positive cerebrospinal fluid, and positive serum, respectively. CONCLUSIONS Our findings show that anti-NR2 antibody in neuropsychiatric lupus serum can damage the BBB and enter the brain.


Nanoscale electronic order in iron pnictides.
The charge distribution in RFeAsO1-xFx (R=La,Sm) iron pnictides is probed using As nuclear quadrupole resonance. Whereas undoped and optimally doped or overdoped compounds feature a single charge environment, two charge environments are detected in the underdoped region. Spin-lattice relaxation measurements show their coexistence at the nanoscale. Together with the quantitative variations of the spectra with doping, they point to a local electronic order in the iron layers, where low- and high-doping-like regions would coexist. Implications for the interplay of static magnetism and superconductivity are discussed.


[Cadmium distribution in tissues and Na,K-ATPase activity of the skin of the frog Rana temporaria in different routes of cadmium uptake by the body].
Accumulation, distribution of Cd in tissues and its toxic effect depend of the mode of uptake of Cd by the organism. Subcutaneous injections of Cd (0.12-0.24 mg/100 g per day) for 10 days resulted in a significant accumulation of Cd in the liver and kidney. No death cases were observed. Addition of CdCl2 to the aquatic environment (more than 0.002%) caused acute toxic effect on frogs. Within 10 days, significant amounts of Cd were found in the skin, especially in its outside layers, small amounts were found in the kidney and liver. High external concentrations of CdCl2 (0.005%) inhibited the activity of Na,K-ATPase in the skin epithelium. It is suggested that adaptive detoxication of cadmium in the liver and kidney operating in mammals, is ineffective in amphibians provided cadmium uptake occurs via the skin.


Broiler performance and intestinal alterations when fed drug-free diets.
A study was carried out to investigate the effects of a drug-free feeding program on broiler performance and intestinal morphology. Chicks vaccinated against coccidia were randomly assigned to 4 dietary treatments: 1) negative control (NC), basal diet; 2) positive control (PC), diet 1 + Lincomycin; 3) program 1 (PG1); diet 1 + Bio-Mos, Vegpro, MTB-100, Acid Pak 4-Way, and All-Lac XCL; 4) and program 2 (PG2), diet 1 + Bio-Mos and All-Lac XCL, each of which were assigned to 13 pens (48 birds in each of 52 pens). Growth traits (BW, feed intake, yield, mortality, BW gain, and feed conversion rate) were obtained through 49 d. At d 14, 3 chicks per pen were challenged with coccidia. Segments of duodenum, ileum, and ceca were removed to measure intestinal morphology at d 14, 28, 35, and 49. Final BW gain of broilers on PC (2.736 kg) was numerically higher than those for NC (2.650 kg). Cumulative feed conversion rate at d 49 was improved (P < 0.05) in birds consuming PC and PG2 compared with NC. Overall, mortality was higher for birds consuming the NC (P < 0.05) than the PC, PG1, and PG2 diets. Interaction of dietary treatments with age and age alone were evident (P < 0.0001) for morphology of duodenum, ileum, and ceca. Lamina propria in ceca was thicker (P < 0.008) in broilers consuming the NC than PG1 and PG2 diets. The results of this study indicated that feeding birds without growth promoters resulted in higher mortality and decreased growth performance than did feeding a diet with an antibiotic, and the combination of Bio-Mos and All-Lac XCL helped to reduce negative effects.


Mycophenolate mofetil added to immunosuppression after liver transplantation--first results.
Mycophenolate mofetil (MMF) has been used successfully as an immunosuppressive agent after kidney and heart transplantation, but experience with MMF after liver transplantation is still limited. Between August 1995 and January 1996, we treated 20 patients with MMF after orthotopic liver transplantation in an open, prospective study. Five out of eight patients with acute rejection and one patient with early chronic rejection showed a complete response after MMF was added to the immunosuppression. Three patients with chronic rejection did not improve, one died, and two have stable graft function at present. In eight patients who suffered from toxicity, a reduction in the dosage of tacrolimus was attempted with simultaneous MMF therapy. One patient died due to multiple organ failure. Liver function improved completely in one other patient, and partially in three patients after adding MMF. In the remaining three patients, a reduced dosage of tacrolimus or cyclosporin, together with MMF, reduced toxicity, not significantly. In conclusion, MMF appears to be a safe and potentially useful adjuvant immunosuppressive agent for rescue and maintenance therapy.


An analysis of the origins of a cooperative binding energy of dimerization.
The cooperativity between binding of cell wall precursor analogs (ligands) to and antibiotic dimerization of the clinically important vancomycin group antibiotics was investigated by nuclear magnetic resonance. When dimerization was weak in the absence of a ligand, the increase in the dimerization constant in the presence of a ligand derived largely from changes associated with tightening of the dimer interface. When dimerization was strong in the absence of a ligand, the increase in the dimerization constant in the presence of a ligand derived largely from changes associated with tightening of the ligand-antibiotic interface. These results illustrate how, when a protein has a loose structure, the binding energy of another molecule to the protein can derive in part from changes occurring within the protein.


Effect of TRPC6 knockdown on puromycin aminonucleoside-induced podocyte injury.
This study was aimed to construct eukaryotic expression vectors carrying the small hairpin RNA (shRNA) targeting TRPC6 gene and investigate the effect of TRPC6 knockdown on puromucin aminonucleoside (PAN)-induced podocyte injury. Two DNA sequences containing the small hairpin structure targeting TRPC6 were designed, synthesized and then inserted into the green fluorescence protein (GFP)-contained plasmids (pGC) to establish the plasmids pGCsi-TRPC6A and pGCsi-TRPC6B. Plasmids expressing scrambled shRNA were used as negative control and named pGCsi-NC. These plasmids were transfected into a conditionally immortalized murine podocyte cell line by using liposome. Flow cytometry was used to examine the transfection efficiency. TRPC6 mRNA and protein expression levels were detected by RT-PCR and Western blotting. Cultured podocytes were divided into four groups: control group, PAN treatment group, PAN+TRPC6 shRNA transfected group and PAN+scrambled shRNA transfected group. The paracelluar permeability to BSA was evaluated by Millicell-PCF Inserts and cell viability was measured by the trypan blue assay. Immunofluorescent assay was used to observe the distribution of α-actinin-4 and α-tubulin. The results showed that the transfection efficiency of the shRNA expression vector was about 45%. Expression levels of TRPC6 mRNA and protein were downregulated after transfection with pGCsi-TRPC6A and pGCsi-TRPC6B. Knocking down TRPC6 gene could effectively reverse the PAN-induced increase in the paracelluar permeability to BSA. The distribution of α-actinin-4 and α-tubulin was disrupted after treatment with PAN, which was reversed by knocking down TRPC6 gene. It was concluded that knocking down TRPC6 gene could effectively prevent podocytes from the permeability increase induced by PAN, which may be related to the regulation of podocyte cytoskeleton.


Glycosuria in primary glomerulopathies: prevalence and prognostic significance.
Tubular damage is common in glomerular diseases (GD). Glycosuria is a marker of tubular dysfunction and may be used to detect tubular lesion and CKD progression. The aim of this study was to evaluate the prevalence and prognostic value of glycosuria at the time of diagnosis in primary glomerulopathies (PG). We conducted a 24-month retrospective study in patients diagnosed with PG in our center between 2009 and 2020. We excluded diabetic patients, use of SGLT2 inhibitors, transplant patients, and secondary GD. Patients were divided in two groups according to their glycosuria status at diagnosis. We studied 115 patients. Global prevalence of glycosuria was 10% (n=11) and membranous nephropathy (MN) had the highest prevalence (n=5, 17.9%). We found that patients with glycosuria had higher serum creatinine (2.4 vs. 1.2 mg/dL, p=0.030), higher albuminuria (4.8 vs. 1.9 g/g, p=0.004), and lower serum albumin (2.3 vs. 3.2 g/dL, p=0.021). We did not find association with histological prognostic factors. At the end of follow-up, patients with glycosuria had higher prevalence of the composite outcome of stage 5D CKD or 50% increase in basal SCr (45.5% vs. 17.3%, p=0.037). In patients with MN, results were similar but we were able to find an association of glycosuria with more severe interstitial fibrosis and tubular atrophy (25.0 vs. 0.0 %, p=0.032). Ten percent of our patients with PG have glycosuria. Glycosuria at the time of diagnosis was associated with more severe clinical presentation and worst renal outcome. The association with higher albuminuria suggests that tubular function has an impact on the severity and outcomes of PG.


The Nexus Between Nutrition and Early Childhood Development.
This article looks at both nutrition and early childhood stimulation interventions as part of an integrated life cycle approach to development. We build on recent systematic reviews of child development, which are comprehensive in regard to what is currently known about outcomes reported in key studies. We then focus particularly on implementation, scaling, and economic returns, drawing mainly on experience in low- and middle-income countries where undernutrition and poor child development remain significant public health challenges with implications across the life course.


SERI surgical scaffold, prospective clinical trial of a silk-derived biological scaffold in two-stage breast reconstruction: 1-year data.
SERI Surgical Scaffold is a long-term bioresorbable silk-derived biological scaffold developed to provide soft-tissue support and repair. SURE-001 (ClinicalTrials.gov identification no. NCT01256502) is a prospective, single-arm study in the United States of patients undergoing two-stage, implant-based breast reconstruction using SERI. A total of 139 patients were enrolled and will be followed for 2 years; in this article, the authors report interim data on 71 patients followed for 1 year. Investigator satisfaction scores (mean ± SD) at 6 and 12 months were 9.2 ± 0.98 and 9.4 ± 0.91, respectively (10 = very satisfied). SERI was rated easy/very easy to use in 98 percent or more of cases across five categories in stage I surgery. Patient satisfaction with the treated breast(s) (mean ± SD) was higher at 6 (4.3 ± 0.87; 5 = very satisfied) and 12 months (4.5 ± 0.82) compared with screening (3.6 ± 1.09; p < 0.0001). Key complication rates (per breast) were tissue necrosis (6.7 percent), seroma (5.7 percent), hematoma (4.8 percent), implant loss (3.8 percent), capsular contracture (1.9 percent), and breast infection (1.0 percent). None were attributed to SERI by the investigators. In 13 patients (14 breasts) who underwent unplanned radiation therapy, one complication was reported. In this interim report, high levels of investigator and patient satisfaction, and ease of use of SERI were reported. Prospectively collected complication rates were similar to those reported in primarily retrospective studies of two-stage, implant-based breast reconstructions using other implantable soft-tissue support materials such as acellular dermal matrices. Therapeutic, IV.


Gene flow in Argentinian sunflowers as revealed by genotyping-by-sequencing data.
Gene flow can have several different applied consequences, ranging from extinction to the escape of transgenes to the evolution of weedy or invasive lineages. Here, we describe patterns of hybridization and gene flow involving domesticated and wild sunflowers in Argentina. To address the risks of introgression of variants from the cultivated sunflower into invasive wild Helianthus, we used genotyping-by-sequencing (GBS) to genotype 182 samples from 11 sites in Argentina, along with previously published data from samples from the native range (North America), to determine the native source populations of the Argentinian samples and to detect admixture. We unexpectedly discovered two distinctive forms of H. petiolaris in Argentina, one from H. petiolaris subsp. petiolaris as expected, but the other from an unknown source. Extensive admixture was observed among Argentinian sunflowers, largely confirming phenotypic predictions. While many hybrids are F1s, there were signals consistent with introgression from the domesticated sunflower into H. petiolaris. Whether this introgression is incidental or a causal driver of invasiveness is not yet clear, but it seems likely that genes found in the domesticated sunflower genome (whether engineered or not) will quickly find their way into wild Argentinian sunflower populations.


Blood coagulation during and after normovolemic hemodilution in elective surgery.
Hemodilution might extend the process of coagulation depending on three possible factors, namely 1) decreased concentration of clotting factors, 2) increased capillary flow and 3) specific actions of the dilutional agent on the clotting mechanism. In two randomized groups of patients undergoing hip arthroplasty preoperative normovolemic hemodilution with dextran 70 was compared to conventional bank blood transfusion. Coagulation parameters, blood loss and incidence of postoperative thromboembolism were studied. Minor reductions of the individual coagulation factors in the immediate postoperative course as well as a protracted APT-time were demonstrated in the hemodiluted group, but total surgical blood loss was not influenced by the hemodilution technique. The frequency of postoperative deep venous thrombosis did not differ between the groups, but there was a significant reduction in the number of silent pulmonary emboli in the hemodilution group.


Protein kinase A-mediated phosphorylation of the Broad-Complex transcription factor in silkworm suppresses its transcriptional activity.
The insect-specific transcription factor Broad-Complex (BR-C) is transcriptionally activated by the steroid 20-hydroxyecdysone (20E) and regulates the expression of many target genes involved in insect growth and development. However, although the transcriptional regulation of BR-C proteins has been well studied, how BR-C is regulated at post-transcription and -translation levels is poorly understood. To this end, using liquid chromatography-tandem mass spectrometry analysis, we identified residue Ser-186 as a phosphorylation site of BR-C in silkworm. Site-directed mutagenesis and treatment with specific kinase activators and inhibitors indicated that the Ser-186 residue in silkworm BR-C is phosphorylated by protein kinase A (PKA). Immunostaining assays disclosed that PKA-mediated phosphorylation of silkworm BR-C has no effect on its nuclear import. However, luciferase reporter analysis, electrophoretic mobility shift assays, and chromatin immunoprecipitation revealed that the PKA phosphorylation event suppresses the transcriptional activation of silkworm BR-C target genes and that this inhibition was caused by repression of BR-C binding to its DNA targets. Of note, both in vitro and ex vivo experiments disclosed that a continuous 20E signal inhibits the PKA-mediated BR-C phosphorylation and also the cAMP/PKA pathway, indicating that 20E's inhibitory effect on PKA-mediated phosphorylation of silkworm BR-C contributes to maintaining BR-C transcriptional activity. In conclusion, our findings indicate that PKA-mediated phosphorylation inhibits silkworm BR-C activity by interfering with its binding to DNA and that 20E signaling relieves PKA-mediated phosphorylation of BR-C, thereby maintaining its transcriptional activity.


The effects of temperature, pH and growth rate on secondary metabolism in Streptomyces thermoviolaceus grown in a chemostat.
Streptomyces thermoviolaceus was grown in a chemostat under conditions of glutamate limitation. The effects of growth rate on production of the antibiotic granaticin, extracellular protein and protease activity as components of secondary metabolism were studied at 37, 45 and 50 degrees C. The amount of each secondary metabolite synthesized was highly dependent on growth rate and temperature. Granaticin yields were highest at growth rates of 0.1 to 0.15 h-1 at 37 degrees C, 0.175 h-1 at 45 degrees C and 0.045 h-1 at 50 degrees C. Protease activity of culture supernatants responded to low nutrient concentration and/or low growth rate. Measurements of extracellular protein revealed complex changes in amount which were dependent on growth rate and temperature. At 45 degrees C and a growth rate of 0.15 h-1, biomass yield was highest between pH 5.5 to 6.5 whereas granaticin synthesis was low at pH 5.5 and rose to highest values at between pH 6.5 and 7.5.


Increased Steroid Excretion in Children with Extremely Low Birth Weight at a Median Age of 9.8 years.
Events during foetal or early extrauterine life may affect bodily structure and/or functions and even pave the way for adult diseases. To find whether extremely low birth weight (ELBW) infants differ from healthy controls regarding the excretion of steroid metabolites. The study compared 17 female and 10 male ELBW infants, all prepubertal, aged 8-11 years, birth weight <1,000 g, with 27 age- and sex-matched controls. All were healthy at the time of the study. Height, weight and BMI did not differ between the groups. Results were adjusted according to body surface area. 36 urinary steroid metabolites were quantified by gas chromatography-mass spectrometry. In the ELBW girls 33/36 steroid metabolites were higher (19 significantly) than in the controls. All 36 steroid metabolites were higher in the ELBW boys (9 significantly) than in the controls. Sums of mineralocorticoid precursors, metabolites descriptive for cortisol and parameters of adrenal androgen production were significantly higher in ELBW infants (both sexes). Only the sum of the metabolites known to be illustrative for adrenal 11β-hydroxysteroid dehydrogenase activity was not different. Prepubertal ELBW children have an augmented urinary excretion of adrenal androgens, cortisol and mineralocorticoid precursors. These findings corroborate and help to explain the link between early-life adversity and subsequent adrenocortical function.


Susceptibility of Clinical Isolates of Burkholderia pseudomallei to a Lipid A Biosynthesis Inhibitor.
Burkholderia pseudomallei is the causative agent of melioidosis, a serious infection associated with high mortality and relapse. Current antimicrobial therapy using ceftazidime (CAZ) is often ineffective. Inhibitors of LpxC, the enzyme responsible for lipid A biosynthesis, have potential antimicrobial activity against several Gram-negative bacteria in vivo, but their activity against B. pseudomallei is unclear. Herein, we investigated the susceptibility of B. pseudomallei clinical isolates to LpxC-4, an LpxC inhibitor, and LpxC-4 in combination with CAZ. Time-kill assays for bactericidal activity were conducted for B. pseudomallei K96243, revealing growth inhibition and bactericidal effect at LpxC-4 concentrations of 2 μg/mL and 4 μg/mL, respectively. No significant synergistic effect was observed with the combination of LpxC-4 and CAZ. LpxC-4 susceptibility was tested on three groups of clinical isolates:1) CAZ- and trimethoprim-sulfamethoxazole (SXT)-susceptible (N = 71), 2) CAZ-resistant (N = 14), and 3) SXT-resistant (N = 23) isolates, by broth microdilution. The minimum concentration of LpxC-4 required to inhibit the growth of 90% of organisms was 2 μg/mL for all isolates. The median minimum inhibitory concentration of both the CAZ/SXT-susceptible and CAZ-resistant groups was 1 μg/mL (interquartile range [IQR] = 1-2 μg/mL), compared with 2 μg/mL (IQR = 2-4 μg/mL) for the SXT-resistant group. Cell morphology was observed after drug exposure by immunofluorescent staining, and a change from rod-shaped to cell wall-defective spherical cells was observed in surviving bacteria. LpxC-4 is a potent bactericidal agent against B. pseudomallei and warrants further testing as a new antibiotic to treat melioidosis.