Photodynamic inactivation of fibroblasts and inhibition of Staphylococcus epidermidis adhesion and biofilm formation by toluidine blue O.
Treating skin and soft tissue infections of severe limb traumas can be challenging. Crucial concerns focus on inhibiting biofilm formation by antibiotic‑resistant bacteria, and preventing scar formation by fibroblastic hyperproliferation. The local use of toluidine blue O (TBO)‑mediated photodynamic therapy (PDT) may be a promising strategy for treating such lesions. The present study used Staphylococcus epidermidis (strain ATCC 35984) to assess the effects of TBO‑PDT on bacterial adherence and biofilm formation, using confocal laser scanning microscopy (CLSM), tissue culture plating (TCP) and scanning electron microscopy (SEM). Primary human fibroblast cells were used to evaluate the cytotoxicity of TBO‑PDT using the 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide (MTT) assay and CLSM. Six different treatment groups were investigated: Medium only [tryptone soy broth (TSB) or Dulbecco's modified Eagle's medium (DMEM)]; red light control (light dose, 30 J/cm2); TBO group (50 mM TBO); TBO‑PDT1 (TBO irradiated with 10 J/cm2); TBO‑PDT2 (TBO irradiated with 20 J/cm2); and TBO‑PDT3 (TBO irradiated with 30 J/cm2). The results of the S. epidermidis adhesion assay indicated that the TSB, light and TBO groups exhibited significant bacterial adherence, compared with the TBO‑PDT groups. Analysis of biofilm formation revealed significant light dose‑dependent differences between the TBO‑PDT groups and the TSB, light, and TBO groups. Furthermore, SEM indicated fewer colony masses in the TBO‑PDT groups compared with the control groups. The MTT assay for fibroblastic cell toxicity demonstrated ~1.1, 4.6, 14.5, 29.7 and 43.4% reduction in optical density for the light, TBO, TBO‑PDT1, TBO‑PDT2 and TBO‑PDT3 groups, respectively, compared with the DMEM control group. There was no difference in toxicity between the light and control groups, however, there were significant differences among the TBO‑PDT groups. Finally, alterations in fibroblast morphology and cell spreading were revealed by CLSM, following TBO‑PDT treatment. TBO‑PDT inhibited bacterial adhesion and biofilm formation, and exhibited significant cytotoxic effects on human fibroblasts. These results indicate that the local use of TBO‑PDT in limb lesions may be a useful treatment method for inhibiting bacterial biofilm formation and fibroblastic hyperproliferation, which may prevent infectious hypertrophic scar formation.


Mental practice: a promising restorative technique in stroke rehabilitation.
Upper limb hemiparesis is a common, yet debilitating, result of stroke. It has long been known that mental practice, when combined with physical practice, improves motor learning and performance. Recent studies also indicate that massed use of the affected arm results in cortical reorganizations and correlative functional improvements. During mental practice, there are widespread activations of neural and muscular mechanisms as if the arm were actually being used. This article introduces mental practice as a form of massed practice, reviews the bases for mental practice as a potent restorative technique, and presents data suggesting mental practice as a restorative technique for upper limb hemiparesis.


Molecular architecture of thin films fabricated via physical vapor deposition and containing a poly(azo)urethane.
Organic thin films are widely applied as transducers in devices whose performance is determined by the optical and electrical properties of the films. In this context, the molecular architecture of the thin films plays an important role. In this work we report the fabrication and characterization of a poly(azo)urethane synthesized fixing CO2 in bis-epoxide followed by a copolymerization reaction with an azodiamine without using isocyanate. The poly(azo)urethane thin films were fabricated by physical vapor deposition (PVD) technique using vacuum thermal evaporation. The molecular architecture of the PVD films was investigated under control growth at nanometer level of thickness, as well as the surface morphology at micro and nanometer scales and the molecular organization. The thermal stability of the poly(azo)urethane molecules, which is a challenge in itself considering the thermal evaporation process, was followed by thermogravimetric analysis (TG) and also by both Fourier transform infrared absorption (FTIR) and ultraviolet-visible (UV-vis) absorption spectroscopies. The UV-vis absorption spectra showed a linear growth of the absorbance of the PVD films with the mass thickness measured by a quartz crystal balance. A random distribution of the poly(azo)urethane molecules in the PVD films was revealed by FTIR spectra. The film morphology was investigated at microscopic level combining chemical and topographical information through micro-Raman technique. At nanoscopic scale, the morphology was investigated by atomic force microscopy (AFM) for films fabricated using distinct evaporation rates. As a proof of principle (for potential applications), the film luminescence was measured over a wide range of temperature. Interestingly, an unusual increase of fluorescence intensity was observed at +150 degrees C after a monotonic decrease from -150 degrees C.


Neurovascular coupling and cerebrovascular hemodynamics are modified by exercise training status at different stages of maturation during youth.
Neurovascular coupling (NVC) is mediated via nitric oxide signaling, which is independently influenced by sex hormones and exercise training. Whether exercise training differentially modifies NVC pre- versus postpuberty, where levels of circulating sex hormones will differ greatly within and between sexes, remains to be determined. Therefore, we investigated the influence of exercise training status on resting intracranial hemodynamics and NVC at different stages of maturation. Posterior and middle cerebral artery velocities (PCAv and MCAv) and pulsatility index (PCAPI and MCAPI) were assessed via transcranial Doppler ultrasound at rest and during visual NVC stimuli. N = 121 exercise-trained (males, n = 32; females, n = 32) and untrained (males, n = 28; females, n = 29) participants were characterized as pre (males, n = 33; females, n = 29)- or post (males, n = 27; females, n = 32)-peak height velocity (PHV). Exercise-trained youth demonstrated higher resting MCAv (P = 0.010). Maturity and training status did not affect the ΔPCAv and ΔMCAv during NVC. However, pre-PHV untrained males (19.4 ± 13.5 vs. 6.8 ± 6.0%; P ≤ 0.001) and females (19.3 ± 10.8 vs. 6.4 ± 7.1%; P ≤ 0.001) had a higher ΔPCAPI during NVC than post-PHV untrained counterparts, whereas the ΔPCAPI was similar in pre- and post-PHV trained youth. Pre-PHV untrained males (19.4 ± 13.5 vs. 7.9 ± 6.0%; P ≤ 0.001) and females (19.3 ± 10.8 vs. 11.1 ± 7.3%; P = 0.016) also had a larger ΔPCAPI than their pre-PHV trained counterparts during NVC, but the ΔPCAPI was similar in trained and untrained post-PHV youth. Collectively, our data indicate that exercise training elevates regional cerebral blood velocities during youth, but training-mediated adaptations in NVC are only attainable during early stages of adolescence. Therefore, childhood provides a unique opportunity for exercise-mediated adaptations in NVC.NEW & NOTEWORTHY We report that the change in cerebral blood velocity during a neurovascular coupling task (NVC) is similar in pre- and postpubertal youth, regardless of exercise-training status. However, prepubertal untrained youth demonstrated a greater increase in cerebral blood pulsatility during the NVC task when compared with their trained counterparts. Our findings highlight that childhood represents a unique opportunity for exercise-mediated adaptations in cerebrovascular hemodynamics during NVC, which may confer long-term benefits in cerebrovascular function.


Excess of cancers in Europe: a study of eleven major cancers amenable to lifestyle change.
Worldwide an estimated 11 million cancer cases were diagnosed in 2002, one quarter being in Europe. We estimated the potential in avoidable numbers and proportions of 11 cancers amenable to prevention (cancers of the oral cavity, oesophagus, stomach, colorectal, pancreas, laryngeal, lung, female breast, endometrium, kidney and bladder) in 28 European countries. We assumed that the aggregated rate of 3 countries with lowest incidence to be attainable throughout Europe. The difference between the age- and gender-specific national cancer incidence rates and the lowest rate observed in 2002 was determined and defined as "avoidable." Of the 1.4 million adult cases of selected cancers and countries within our study, 363,000 (59%) cancers in males and 326,000 (45%) cancers in females were hypothetically avoidable. Among men, the proportion was largest in Hungary (77%) and among women, in Belgium (54%). Assuming that differences in cancer incidence are not attributable to genetic susceptibility or diagnostic activity, about 50% of all cases of these 11 cancers could be potentially avoided, especially by decreased smoking among men. Interventions directed at reducing smoking, obesity and alcohol use as well as increasing physical activity and fruit and vegetable intake are necessary to attain lower incidence rates. It is important to recognize that the actual preventable cancer by eliminating currently known risk factors is somewhat less than we have estimated.


Establishing the Maximum Collectivity in Highly Deformed N=Z Nuclei.
The lifetimes of the first excited 2^{+} states in the N=Z nuclei ^{80}Zr, ^{78}Y, and ^{76}Sr have been measured using the γ-ray line shape method following population via nucleon-knockout reactions from intermediate-energy rare-isotope beams. The extracted reduced electromagnetic transition strengths yield new information on where the collectivity is maximized and provide evidence for a significant, and as yet unexplained, odd-odd vs even-even staggering in the observed values. The experimental results are analyzed in the context of state-of-the-art nuclear density-functional model calculations.


Determination of phagocytosis of 32P-labeled Staphylococcus aureus by bovine polymorphonuclear leukocytes.
A procedure for the measurement of phagocytosis by bovine polymorphonuclear leukocytes (PMN) of 32P-labeled Staphylococcus aureus was modified so that a larger number of samples could be compared in a single run, and smaller volumes of sample, PMN, and 32P-labeled S aureus could be used. Results were highly reproducible, with a coefficient of variation between duplicate determinations of less than or equal to 2%. Lysostaphin was prepared from the supernatant of S staphylolyticus and was compared with a commercially available preparation. Effects of lysostaphin on PMN and influence of incubation media on release of 32P from 32P-labeled S aureus by lysostaphin were examined.


Fully Recyclable Cured Polymers for Sustainable 3D Printing.
The most prevalent materials used in the Additive Manufacturing era are polymers and plastics. Unfortunately, these materials are recognized for their negative environmental impact as they are primarily nonrecyclable, resulting in environmental pollution. In recent years, a new sustainable alternative to these materials has been emerging: Reversible Covalent Bond-Containing Polymers (RCBPs). These materials can be recycled, reprocessed, and reused multiple times without losing their properties. Nonetheless, they have two significant drawbacks when used in 3D printing. First, some require adding new materials every reprinting cycle, and second, others require high temperatures for (re)printing, limiting recyclability, and increasing energy consumption. This study, thus, introduces fully recyclable RCBPs as a sustainable approach for radiation-based printing technologies. This approach enables multiple (re)printing cycles at low temperatures (50 °C lower than the lowest reported) without adding new materials. It involves purposefully synthesized polymers that undergo reversible photopolymerization, composed of a tin-based catalyst. An everyday microwave oven quickly depolymerized these polymers, obtaining complete reversibility.


[A case of renal cell carcinoma in horseshoe kidney].
Horseshoe kidney has a frequency of 1 in 1000 renal paiss. Diseases in this type of kidneys have anatomical particularities that should be taken into account during surgery. We present the case of a renal cell carcinoma in this type of renal fusion.


Genome Copy Number Quantification Revealed That the Ethanologenic Alpha-Proteobacterium Zymomonas mobilis Is Polyploid.
The alpha-proteobacterium Zymomonas mobilis is a promising biofuel producer, based on its native metabolism that efficiently converts sugars to ethanol. Therefore, it has a high potential for industrial-scale biofuel production. Two previous studies suggested that Z. mobilis strain Zm4 might not be monoploid. However, a systematic analysis of the genome copy number is still missing, in spite of the high potential importance of Z. mobilis. To get a deep insight into the ploidy level of Z. mobilis and its regulation, the genome copy numbers of three strains were quantified. The analyses revealed that, during anaerobic growth, the lab strain Zm6, the Zm6 type strain obtained from DSMZ (German Collection of Microorganisms), and the lab strain Zm4, have copy numbers of 18.9, 22.3 and 16.2, respectively, of an origin-adjacent region. The copy numbers of a terminus-adjacent region were somewhat lower with 9.3, 15.8, and 12.9, respectively. The values were similar throughout the growth curves, and they were only slightly downregulated in late stationary phase. During aerobic growth, the copy numbers of the lab strain Zm6 were much higher with around 40 origin-adjacent copies and 17 terminus-adjacent copies. However, the cells were larger during aerobic growth, and the copy numbers per μm3 cell volume were rather similar. Taken together, this first systematic analysis revealed that Z. mobilis is polyploid under regular laboratory growth conditions. The copy number is constant during growth, in contrast to many other polyploid bacteria. This knowledge should be considered in further engineering of the strain for industrial applications.


The musculoskeletal manifestations of the Coffin-Lowry syndrome.
Coffin-Lowry syndrome (CLS) is a rare genetic disorder characterized by craniofacial abnormalities, mental retardation, short stature, and hypotonia. Patients with CLS may present with multiple musculoskeletal abnormalities. The purpose of this study was to identify and characterize the musculoskeletal findings in 10 patients with CLS. Eight patients presented with thoracolumbar kyphosis or kyphoscoliosis, with a mean Cobb angle of 45 degrees in the coronal plane and 31 degrees of thoracolumbar kyphosis. These may be progressive and difficult to treat, needing early surgical treatment. Close follow-up of the spinal deformities is strongly recommended to document progression. Sixty percent of the patients presented with bilateral flexible and painless planovalgus deformities. Hypoplasia of the ilium and hand deformities are common but do not seem to cause any functional problems. Observation is recommended for these asymptomatic hand, foot, and pelvic findings.


Protective effect of Salvia miltiorrhiza Bunge on 5-fluorouracil-induced oral mucositis.
Oral mucositis is a common side-effect caused by chemotherapy or radiotherapy occurring in the majority of cancer patients and is characterized by inflammation and ulcers in the oral mucosa. In the present study, we examined the protective effects of Salvia miltiorrhiza Bunge (SM) on oral mucositis induced by 5-fluorouracil (5-FU) in human pharyngeal cells and golden Syrian hamsters. We investigated the proliferation and antioxidant abilities of SM using MTT, 2-diphenyl-1-picrylhydrazyl (DPPH) and reactive oxygen species (ROS) assays in vitro. Additionally, TUNEL assay was performed, and the expression levels of nuclear factor-κB (NF-κB), caspase-3 and proinflammatory cytokines were assessed by immunoblotting. The results showed that SM increased the cell proliferation rate in human pharyngeal cells up to 128.97±9.7% compared with this rate in the untreated cells and exerted protective effects on mucosal injury caused by 5-FU treatment. In addition, all concentrations of SM increased DPPH scavenging ability and blocked ROS generation in the treated cells. Taken together, following SM treatment, expression of NF-κB and cleaved caspase-3 were significantly decreased followed by inhibition of cell death. These data suggest that SM could be used for the prevention and treatment of oral mucositis caused by cancer therapies.


Influence of pH on the self-assembly of diphenylalanine peptides: molecular insights from coarse-grained simulations.
Nanostructures fabricated from peptide self-assemblies are attracting increasing attention owing to their possible applications in biology and nanotechnology. A known example is an aromatic dipeptide (diphenylalanine, FF) which is extracted from Alzheimer's β-amyloid polypeptide as the core recognition motif for molecular self-assembly. Many studies have been carried out to organize FF peptides into various functional ordered nanostructures. For potential applications of self-assembled FF-based nanomaterials, it becomes important to consider some influencing factors (e.g., solvents, peptide concentrations, pH, temperature, etc.) on the self-assembly process. Among these factors, the effect of pH on the self-assembly process of FF peptides into assembled nanostructures through simulation studies is the main focus of the present work. In the current study, we have investigated the assembly pathway of 1000 FF peptides and qualitatively evaluated the morphological changes of FF-based nanostructures at different pH values by performing extensive coarse-grained molecular dynamics (CG-MD) simulations. Structural analyses suggest that FF peptides can spontaneously assemble into nanotubes with different shapes under acidic, neutral and basic conditions. Based on the analysis of FF nanostructure formation pathways in different pH solutions, the self-assembly of the nanotube involves the aggregation of molecules to form a bilayer, the curling of a bilayer to form a vesicle and the transformation of a vesicle into a tubular structure. It is noted that a flat hollow columnar structure is observed as a special intermediate state during the transformation process of a vesicle-like to a tube-like structure. Energetic analysis suggests that the aggregation of FF peptides is driven by the vdW interactions but the aggregation shape is mainly affected by the electrostatic interactions. Overall, this study provides further understanding of the self-assembly behavior of aromatic short peptide derivatives in different pH solutions.


The Rat Genome Database 2013--data, tools and users.
The Rat Genome Database (RGD) was started >10 years ago to provide a core genomic resource for rat researchers. Currently, RGD combines genetic, genomic, pathway, phenotype and strain information with a focus on disease. RGD users are provided with access to structured and curated data from the molecular level through the organismal level. Those users access RGD from all over the world. End users are not only rat researchers but also researchers working with mouse and human data. Translational research is supported by RGD's comparative genetics/genomics data in disease portals, in GBrowse, in VCMap and on gene report pages. The impact of RGD also goes beyond the traditional biomedical researcher, as the influence of RGD reaches bioinformaticians, tool developers and curators. Import of RGD data into other publicly available databases expands the influence of RGD to a larger set of end users than those who avail themselves of the RGD website. The value of RGD continues to grow as more types of data and more tools are added, while reaching more types of end users.


Electron diffraction study of the space group of Bi5Ti3FeO15 multiferroic ceramic.
Bi5Ti3FeO15 (pentabismuth trititanium iron pentadecaoxide), which is a multiferroic four-layer Aurivillius phase compound, has received much attention in recent years. However, three mutually inconsistent orthorhombic space groups, i.e. A21am, Fmm2 and Pnn2, have been reported for the room-temperature phase of Bi5Ti3FeO15 by X-ray and neutron diffraction investigations. Here, electron diffraction results are presented and discussed for the first time to unambiguously clarify the room-temperature space group of ceramic Bi5Ti3FeO15. It has been found that all the observed reflections from the ceramic agree with those expected in A21am, while the observed reflections 011, 013 and 015 should be forbidden in the case of Fmm2, and no 107 and 109 reflections were observed although allowed for Pnn2. The present study has demonstrated that the space group of Bi5Ti3FeO15 ceramic is A21am rather than Fmm2 or Pnn2, an identification that proved to be a challenge for X-ray diffraction. On the basis of the space group A21am, the lattice parameters of the Bi5Ti3FeO15 ceramic were calculated from its X-ray diffraction data.


Quantitative analysis of nanometer anatase-rutile mixtures via experimentation and theoretical calculation.
In this paper, we developed a method based on X-ray diffractometry for determining the weight ratio of nanometer anatase to rutile or their relative amounts in TiO2 polymorphs, and corresponding formulas for such determination were put forward. The very key constant K in these formulas has been specially evaluated by experimentalizing nanometer polymorphs of titanium dioxide. Furthermore, a theoretical value of K was obtained through detailed theoretical calculation based on X-ray powder diffraction theory. The K value concluded from experimentation coincides well with that yielded from theoretical calculation, thus the validity and reliability of the experimental K value has been further confirmed. This coincidence may also suggest the applicability of X-ray powder diffraction theory to nanometer crystals. With this method, the relative amounts of nanometer anatase and rutile in their mixtures or the weight ratio of nanometer anatase to rutile in any a mixture can be easily determined only upon a XRD test.


Simplified assessment of asphyxia at birth.
A simplified clinical method of assessment of the baby soon after birth was evaluated and compared with the cord blood pH and 1 min Apgar score. The assessment recorded cry, respiration, colour, activity, reflex response, and cord pulsation. Sixty-two neonates born at term by normal vaginal delivery with 1 min Apgar score of 0-6 (n = 31) and 7-10 (n = 31) were studied. Linear correlation coefficient (r) and Goodman's correlation coefficient (gamma) were used for statistical comparison. A combination of cry, colour and activity showed the best correlation with cord blood pH (r = 0.3842, P < 0.01: gamma = 0.7138) and 1 min Apgar score (r = 0.7768, P < 0.01, gamma = 0.9393) and was as useful as the combination of all the six clinical parameters. Illiterate traditional birth attendants can be safely taught to use cry, colour, and activity for assessing baby's condition at birth.


Photon- and Singlet-Oxygen-Induced Cis-Trans Isomerization of the Water-Soluble Carotenoid Crocin.
Studying the cis-trans isomerization process in crocin (CR), one of the few water-soluble carotenoids extracted from saffron, is important to better understand the physiological role of cis-carotenoids in vivo and their potential as antioxidants in therapeutic applications. For that, cis-trans isomerization of both methanol- and water-dissolved CR was induced by light or thermally generated singlet oxygen (1O2). The kinetics of molecular concentrations were monitored by both high-performance liquid chromatography (HPLC) and non-destructive spectrophotometric methods. These last made it possible to simultaneously follow the cis-trans isomerization, the possible bleaching of compounds and the amount of thermally generated 1O2. Our results were in accordance with a comprehensive model where the cis-trans isomerization occurs as relaxation from the triplet state of all-trans- or 13-cis-CR, whatever is the way to populate the CR triplet state, either by photon or 1O2 energy transfer. The process is much more (1.9 to 10-fold) efficient from cis to trans than vice versa. In H2O, a 1O2-induced bleaching effect on the starting CR was not negligible. However, the CR "flip-flop" isomerization reaction could still occur, suggesting that this process can represent an efficient mechanism for quenching of reactive oxygen species (ROS) in vivo, with a limited need of carotenoid regeneration.


Risk factors of pelvic lymph node metastasis in cervical adenocarcinoma following radical hysterectomy and pelvic lymphadenectomy.
Lymph node metastasis is the most important prognostic factor in cervical cancerpatients. However, most of the available knowledge about risk factors of pelvic nodal metastasis in cervical cancer has come from studies in which the majority of patients had the squamous cell carcinoma (SCC) subtype. To determine the risk factors of pelvic lymph node metastasis in early-stage cervical adenocarcinoma (AC) patients following radical hysterectomy and bilateral pelvic lymphadenectomy. Retrospective reviews were carried out of the medical charts and pathologic slides of 251 patients with cervical AC stage IB1-IIA who underwent radical hysterectomy and bilateral pelvic lymphadenectomy at Rajavithi Hospital from January 1, 2000 to December 31, 2011. The risk factors of pelvic lymph node metastasis were analyzed by multiple logistic regression. Of the 251 patients, pelvic node metastasis in stage IB1-IIA cervical AC was detected in 29 patients (11.6%). Multivariable analysis revealed that clinical stage IB2-IIA (adjusted OR 3.4, 95%CI 1.2-9.7), tumor size more than 2 cm (adjusted OR 3.5, 95%CI 1.1-11.8), and positive lymphovascular invasion (LVSI) (adjusted OR 55.5, 95%CI 7.2-427.6) were significantly associated with pelvic nodal metastasis. Early-stage cervical AC patients with no risk factor, one risk factor other than LVSI, LVSI factor alone, two risk factors, and three risk factors were identified as having pelvic nodal metastasis in 0%, 2.3%, 9.1%, 29.1% and 58.8% of cases respectively. Clinical stage IB2-IIA, tumor size of more than 2 cm, and positive LVSI were significant risk factors for pelvic nodal metastasis in early-stage cervical AC patients. Those with no risk factors were not found to have pelvic nodal metastasis and might be candidates for less radical surgery, whereas patients with the presence of LVSI and/or 2 other risk factors were found to be at high risk of pelvic node metastasis and might benefit from extensive lymphadenectomy and adjuvant therapy.


Insufficient documentation of the hemodialysis prescription: delaying the definition of adequate dialysis.
We analyzed 953 articles published in three established nephrology journals to select those pertaining to treatment with hemodialysis. A total of 152 papers were selected on the criteria that the reported results might have been affected by variation in the dialysis prescription. Only a small proportion of the papers studied (11.8%) detailed the complete dialysis prescription including type of dialyzer, blood flow rate, and duration and frequency of dialysis. A surprising 29% of papers provided no details of the dialyzer employed. Other potentially important variables such as the type of blood tubing or method of water treatment used were rarely mentioned. Comparative studies of the course and outcome of hemodialysis regimens require specification of exactly how dialysis was performed. It is suggested that manuscripts provide this information as a condition of suitability for publication.


Relative Sinus Bradycardia: An Unexpected Finding in Preeclampsia With Acute Pulmonary Edema.
Preeclampsia is a multifactorial pregnancy-specific syndrome, which can result in significant alterations in cardiovascular hemodynamics. We report an observation of unexpected relative bradycardia in a previously healthy woman who presented with postpartum preeclampsia and acute pulmonary edema. We observed an increase in heart rate following normalization of blood pressure, which suggested that the initial slower than expected heart rate may reflect a baroreceptor response to hypertension. Whether this finding should be regarded as a severe aspect of the disease spectrum needs further study.


Oral, intestinal, and skin bacteria in ventral hernia mesh implants.
In ventral hernia surgery, mesh implants are used to reduce recurrence. Infection after mesh implantation can be a problem and rates around 6-10% have been reported. Bacterial colonization of mesh implants in patients without clinical signs of infection has not been thoroughly investigated. Molecular techniques have proven effective in demonstrating bacterial diversity in various environments and are able to identify bacteria on a gene-specific level. The purpose of this study was to detect bacterial biofilm in mesh implants, analyze its bacterial diversity, and look for possible resemblance with bacterial biofilm from the periodontal pocket. Thirty patients referred to our hospital for recurrence after former ventral hernia mesh repair, were examined for periodontitis in advance of new surgical hernia repair. Oral examination included periapical radiographs, periodontal probing, and subgingival plaque collection. A piece of mesh (1×1 cm) from the abdominal wall was harvested during the new surgical hernia repair and analyzed for bacteria by PCR and 16S rRNA gene sequencing. From patients with positive PCR mesh samples, subgingival plaque samples were analyzed with the same techniques. A great variety of taxa were detected in 20 (66.7%) mesh samples, including typical oral commensals and periodontopathogens, enterics, and skin bacteria. Mesh and periodontal bacteria were further analyzed for similarity in 16S rRNA gene sequences. In 17 sequences, the level of resemblance between mesh and subgingival bacterial colonization was 98-100% suggesting, but not proving, a transfer of oral bacteria to the mesh. The results show great bacterial diversity on mesh implants from the anterior abdominal wall including oral commensals and periodontopathogens. Mesh can be reached by bacteria in several ways including hematogenous spread from an oral site. However, other sites such as gut and skin may also serve as sources for the mesh biofilm.


Relationships between basal hypothalamic-pituitary-thyroid-axis activity and plasma haptoglobin levels in depression.
The present study was carried out in investigate the relationship between basal hypothalamic-pituitary-thyroid (HPT)-axis function and the acute phase (AP) response in depression. Toward this end, the authors measured serum concentrations of basal thyroid-stimulating-hormone (TSH), free thyroxine (FT4) and free triiodothyronine (FT3), and plasma levels of an AP protein, i.e. haptoglobin (Hp) in 38 depressed in-patients and in 11 normal controls. In depression, basal TSH was significantly and negatively related to Hp values, whereas in normal controls a trend toward a positive correlation between both factors was found. Depressed patients with increased Hp levels (> or = 250 mg/dl) showed significantly lower basal TSH levels than patients with normal Hp levels. No significant correlations were found between Hp plasma levels and either FT4 or FT3 serum concentrations. The results support the hypothesis that lower TSH secretion in major depression may be related to the AP response in that illness, and may constitute an expression of a coordinated neuroendocrine-immune response to nonthyroidal illness.


Clinical Research Progress of BTK Inhibitors in the Treatment of Autoimmune Diseases.
Bruton tyrosine kinase (BTK) is an important protein of the tyrosine kinase family and plays a key role in signal transduction, proliferation, migration, and survival in B lymphocytes. The inhibition of BTK is a promising therapy for various autoimmune diseases (AD) involving abnormal B cell function, such as rheumatoid arthritis (RA), multiple sclerosis (MS), and systemic lupus erythematosus (SLE). This article briefly summarizes the role of BTK in the BCR signaling pathway, the development process of BTK inhibitors, and especially the latest progress of their clinical trials for the treatment of AD.


Forebrain contribution to sympathetic nerve discharge in anesthetized cats.
We tested the hypothesis that the forebrain is responsible for a component of basal sympathetic nerve discharge (SND) in the anesthetized cat. For this purpose integrated postganglionic inferior cardiac SND and blood pressure were measured before and after serial transections of the midbrain at stereotaxic planes A3 and AP0. In the first series of experiments on baroreceptor-denervated cats anesthetized with alpha-chloralose, A3 transection significantly reduced SND to 62 +/- 7% of control. Blood pressure was reduced 33 +/- 4 mmHg. SND and frontal-parietal cortical activity were temporally related prior to A3 transection in many of these animals. Recovery of SND and blood pressure to control levels occurred within 30 min after A3 transection. A second midbrain transection at AP0 failed to affect SND and blood pressure at this time. This observation suggests that the effects of A3 transection were due to the loss of a component of SND of forebrain origin rather than to generalized trauma. The effects of A3 transection were not attributable to cataleptic anesthesia with alpha-chloralose, since SND and blood pressure were significantly reduced by midbrain transection in baroreceptor-denervated cats anesthetized with diallylbarbiturate-urethan. A3 transection also reduced SND and blood pressure in some baroreceptor-innervated cats. Finally, medial diencephalic ablation prevented the effects of A3 transection. We conclude that the forebrain is responsible for a significant component of sympathetic tone in anesthetized cats. Recovery from the loss of this component, however, occurs rapidly even in animals deprived of their compensatory baroreceptor reflex mechanisms.


Oxidative metabolism of glutathione by gamma-glutamyl transpeptidase and peroxisome proliferation: the relevance to hepatocarcinogenesis. A hypothesis.
Much has been learned about the function of glutathione (GSH) and other thiols as antioxidants, radioprotectors and radical scavengers. Recent reports point out that GSH and other thiols are double-edged swords: they induce the formation of free radicals and oxidative damage. Such damage is responsible for most, if not all, genotoxicity of GSH to bacteria, and probably to mammalian cells as well. The activation of GSH to an oxidative mutagen and induction of oxidative damage by GSH are catalysed by gamma-glutamyl transpeptidase (GGT), an enzyme appearing very frequently in enzyme-altered foci in livers of rodents, shortly after their exposure to carcinogens. It is proposed here that such GGT-dependent oxidative damage may help in the promotion stage in tumourigenesis, and in that its function may be similar to that of peroxisome proliferators as promotors of hepatocarcinogenesis.


Anomalous cytokeratin expression in malignant melanoma: one- and two-dimensional western blot analysis and immunohistochemical survey of 100 melanomas.
Immunohistochemical characterization is an accepted method of human cell typing and tumor diagnosis. The differentiation of undifferentiated carcinoma from amelanotic melanoma is usually achieved by demonstration of cytokeratin (CK) intermediate filaments in carcinoma but not in melanoma. In examination of 100 melanomas fixed in formalin or methacarn and frozen tissue sections, we have found CK-immunoreactivity in 2, 8, and 21% of cases, respectively, with multiple anticytokeratin antibodies displaying overlapping antigenic specificities. In addition, we have confirmed the anomalous expression of low molecular weight CK proteins by one- and two-dimensional gel electrophoresis and immunoblotting of extracts of an immunohistochemically positive case. This latter finding indicates that CK staining in melanomas reflects the presence of authentic CK peptides and is not an artefact induced by fixation or cross-reacting antibodies. These observations have direct implications for the application of immunohistochemistry to the present practice of diagnostic surgical pathology. The anomalous CK expression by melanoma limits the diagnostic reliability of immunohistochemically demonstrated CK alone to indicate a diagnosis of carcinoma, without the concomitant detection of additional tumor-associated antigens. The finding of anomalous CK expression only in metastatic or recurrent melanomas raises an interesting question of possible association with tumor progression.


Source localization of auditory evoked potentials after cochlear implantation.
Little is known about how the auditory cortex adapts to artificial input as provided by a cochlear implant (CI). We report the case of a 71-year-old profoundly deaf man, who has successfully used a unilateral CI for 4 years. Independent component analysis (ICA) of 61-channel EEG recordings could separate CI-related artifacts from auditory-evoked potentials (AEPs), even though it was the perfectly time-locked CI stimulation that caused the AEPs. AEP dipole source localization revealed contralaterally larger amplitudes in the P1-N1 range, similar to normal hearing individuals. In contrast to normal hearing individuals, the man with the CI showed a 20-ms shorter N1 latency ipsilaterally. We conclude that ICA allows the detailed study of AEPs in CI users.


Multiparametric Biosensors for Characterizing Extracellular Vesicle Subpopulations.
Extracellular vesicles (EVs) are an important intercellular communication conduit for cells that have applications in precision therapy and targeted drug delivery. Small EVs, or exosomes, are a 30-150 nm phospholipid-encased subpopulation of EVs that are particularly difficult to characterize due to their small size and because they are difficult to isolate using conventional methods. In this review, we discuss some recent advances in exosome isolation, purification, and sensing platforms using microfluidics, acoustics, and size exclusion chromatography. We discuss some of the challenges and unanswered questions with respect to understanding exosome size heterogeneity and how modern biosensor technology can be applied to exosome isolation. In addition, we discuss how some advancements in sensing platforms such as colorimetric, fluorescent, electronic, surface plasmon resonance (SPR), and Raman spectroscopy may be applied to exosome detection in multiparametric systems. The application of cryogenic electron tomography and microscopy to understanding exosome ultrastructure will become vital as this field progresses. In conclusion, we speculate on some future needs in the exosome research field and how these technologies could be applied.


Frequent Pattern Mining in Continuous-Time Temporal Networks.
Networks are used as highly expressive tools in different disciplines. In recent years, the analysis and mining of temporal networks have attracted substantial attention. Frequent pattern mining is considered an essential task in the network science literature. In addition to the numerous applications, the investigation of frequent pattern mining in networks directly impacts other analytical approaches, such as clustering, quasi-clique and clique mining, and link prediction. In nearly all the algorithms proposed for frequent pattern mining in temporal networks, the networks are represented as sequences of static networks. Then, the inter- or intra-network patterns are mined. This type of representation imposes a computation-expressiveness trade-off to the mining problem. In this paper, we propose a novel representation that can preserve the temporal aspects of the network losslessly. Then, we introduce the concept of constrained interval graphs ( CIGs). Next, we develop a series of algorithms for mining the complete set of frequent temporal patterns in a temporal network data set. We also consider four different definitions of isomorphism for accommodating minor variations in temporal data of networks. Implementing the algorithm for three real-world data sets proves the practicality of the proposed approach and its capability to discover unknown patterns in various settings.


Morphological plasticity of motor axons in Drosophila mutants with altered excitability.
An anatomical and electrophysiological study of Drosophila mutants has been made to determine the effect of altered electrical activity on the development and maintenance of larval neuromuscular junctions. We examined motor axon terminals of (1) hyperexcitable mutants Shaker (Sh), ether a go-go (eag), Hyperkinetic (Hk), and Duplication of para+ (Dp para+); and (2) mutants with reduced excitability, no action potential (napts) and paralytic (parats 1). Nerve terminals innervating larval body-wall muscles were visualized by using anti-HRP immunocytochemistry, which specifically stains neurons in insect species. In wild-type larvae, motor axon terminals were distributed in a stereotypic fashion. However, in combinations of eag and Sh alleles, the basic pattern of innervation was altered. There was an increase in both the number of higher-order axonal branches over the muscles and the number of varicosities on the neurites. A similar phenomenon was found in the double mutant Hk eag and, to a lesser extent, in Dp para+ and Dp para+ Sh mutants. It is known that at permissive temperature the napts, but not parats 1, mutation decreases excitability of larval motor axons and suppresses the behavioral phenotypes of Sh, eag, and Hk. In the mutant napts (reared at permissive temperature), a slight decrease in the extent of branching was observed. Yet, when combined with eag Sh, napts completely reversed the morphological abnormality in eag Sh mutants. No such reversion was observed in parats 1 eag Sh mutants. The endogenous patterns of electrical activity at the neuromuscular junction were analyzed by extracellular recordings in a semi-intact larval preparation. Recordings from wild-type body-wall muscles revealed rhythmic bursts of spikes. In eag Sh mutants, this rhythmic activity was accompanied by or superimposed on periods of strong tonic activity. This abnormal pattern of activity could be partially suppressed by napts in combination with eag Sh.


Interaction of palmatine with DNA: an environmentally controlled phototherapy drug.
Palmatine is one of the four main protoberberine alkaloids and is largely employed in pharmacy and medicine as a versatile drug with considerable biological activities. More recently, palmatine has been proposed as a promising DNA phototherapy drug, notably due to its ability to produce in situ singlet oxygen only when interacting with DNA. The fine mechanisms of palmatine-DNA interactions as well as its complicated photophysics are not yet fully understood. In this paper, we identify via molecular dynamic techniques two stable interaction modes between palmatine and B-DNA, namely insertion and minor groove binding, whose structural and electronic bases are analyzed and rationalized. These two competitive modes share the same UV-vis signature and estimated binding free energies, and thus they may indeed coexist. By using hybrid quantum mechanics/molecular mechanics protocols coupled to molecular dynamics, we analyze palmatine excited state properties in water solution and in interaction with DNA. The environmentally controlled production of singlet oxygen is thus rationalized in terms of the competition between local and charge-transfer excited states.


Microsomal and cytosolic fractions of guinea pig hepatocytes contain 100-kilodalton GTP-binding proteins reactive with antisera against alpha subunits of stimulatory and inhibitory heterotrimeric GTP-binding proteins.
Guinea pig hepatocytes fractionated by differential centrifugation into plasma membrane-enriched, microsomal, and cytosolic fractions were examined for their content of alpha and beta subunits of heterotrimeric GTP-binding proteins (G proteins) involved in signal transduction. alpha subunits of stimulatory (Gs) and inhibitory (Gi) proteins were detected by immunoblots with antisera reactive with the carboxyl-terminal decapeptide regions of these proteins. Unexpectedly, antisera (including immunopurified) to the alpha subunit but not the beta subunit reacted with a band of 100-kDa proteins in both the microsomal and cytosolic fractions. The immunoreactive 100-kDa proteins are not substrates for ADP-ribosylation catalyzed by pertussis toxin, cholera toxin, or diptheria toxin. Protease digests of the 100-kDa proteins yielded immunoreactive peptides that are distinctly different from those obtained from protease digests of alpha subunits of heterotrimeric G proteins. The 100-kDa protein(s) reactive with antisera to Gi alpha subunit bind to GTP-agarose but not to ATP-agarose. It is concluded that the immunoreactive 100-kDa proteins in microsomal and cytosolic fractions are structurally distinct G proteins from those linked to receptors in the plasma membrane and other G proteins such as elongation factor 2. Conceivably, the 100-kDa proteins represent a new class of G proteins.


Visualizing key hinges and a potential major source of compliance in the lever arm of myosin.
We have determined the 2.3-Å-resolution crystal structure of a myosin light chain domain, corresponding to one type found in sea scallop catch ("smooth") muscle. This structure reveals hinges that may function in the "on" and "off" states of myosin. The molecule adopts two different conformations about the heavy chain "hook" and regulatory light chain (RLC) helix D. This conformational change results in extended and compressed forms of the lever arm whose lengths differ by 10 Å. The heavy chain hook and RLC helix D hinges could thus serve as a potential major and localized source of cross-bridge compliance during the contractile cycle. In addition, in one of the molecules of the crystal, part of the RLC N-terminal extension is seen in atomic detail and forms a one-turn alpha-helix that interacts with RLC helix D. This extension, whose sequence is highly variable in different myosins, may thus modulate the flexibility of the lever arm. Moreover, the relative proximity of the phosphorylation site to the helix D hinge suggests a potential role for conformational changes about this hinge in the transition between the on and off states of regulated myosins.


Auditory imagery shapes movement timing and kinematics: evidence from a musical task.
The role of anticipatory auditory imagery in music-like sequential action was investigated by examining timing accuracy and kinematics using a motion capture system. Musicians responded to metronomic pacing signals by producing three unpaced taps on three vertically aligned keys at the given tempo. Taps triggered tones in two out of three blocked feedback conditions, where key-to-tone mappings were compatible or incompatible in terms of spatial and pitch height. Results indicate that, while timing was most accurate without tones, movements were smaller in amplitude and less forceful (i.e., acceleration prior to impact was lowest) when tones were present. Moreover, timing was more accurate and movements were less forceful with compatible than with incompatible auditory feedback. Observing these effects at the first tap (before tone onset) suggests that anticipatory auditory imagery modulates the temporal kinematics of regularly timed auditory action sequences, like those found in music. Such cross-modal ideomotor processes may function to facilitate planning efficiency and biomechanical economy in voluntary action.


Effect of pressure on proton-coupled electron transfer reactions of seven-coordinate iron complexes in aqueous solutions.
For the first time, the effect of pressure on proton-coupled electron-transfer reactions of two selected seven-coordinate FeIII/II(H2L)(H2O)2 systems [where H2L = 2,6-diacetylpyridine-bis(semicarbazone) and 2,6-diacetylpyridine-bis(semioxamazide), respectively] was examined. The acid-base equilibria of the different Fe(III/II) systems were investigated by spectrophotometric, potentiometric, and electrochemical titrations. On the basis of the obtained species distributions, the pH intervals in which the different protonated forms of the two studied systems exist were defined. In different pH ranges, a different number of protons (from 0 to 3 protons per electron) can be transferred during the redox process, which affects the change in the overall charge on the complexes. For all the different protonation forms of the studied complexes, the change in the redox potentials with pressure was measured and the redox reaction volume was obtained by high-pressure cyclic voltammetry. The results show that in the case of proton-coupled electron transfer, the reaction volume for the neutralization of protons contributes to the overall reaction volume. A linear correlation between Deltaz2 (change in the square of the charge) and the overall reaction volume of the complexes upon reduction, DeltaVcomplex0, was found. The average value of the intrinsic volume change for the selected seven-coordinate iron complexes was estimated from the intercept of the plot of DeltaVcomplex0 versus Deltaz2 to be 9.2 +/- 0.7 cm3 mol(-1). For the combined redox and protonation processes, the data are discussed in terms of linear correlations between Deltaz2 and the redox and neutralization reaction volumes reported in the literature.


Micropatterned substrates for the growth of functional neuronal networks of defined geometry.
The in vitro assembly of neuronal networks with control over cell position and connectivity is a fascinating approach not only for topics in basic neuroscience research but also in diverse applications such as biosensors and tissue engineering. We grew rat embryonic cortical neurons on patterned substrates created by microcontact printing. Polystyrene was used as a cell repellent background, onto which a grid pattern of physiological proteins was applied. We printed laminin and a mixture of extracellular matrix proteins and additionally both systems mixed with polylysine. Attachment of cells to the pattern with high fidelity as well as the formation of chemical synapses between neighboring cells on the pattern could be observed in all four cases, but cell attachment was strongly increased on samples containing polylysine. Neurons grown on patterned substrates had a membrane capacity smaller than that of neurons on homogeneously coated controls, which we attributed to the geometrical restrictions, but did not differ either in resting membrane potential or in the quality of synapses they formed. We therefore believe that the cells attach and differentiate normally on the pattern and form functional, mature synapses following the predefined geometry.


Meeting the needs of new ostomists: a patient evaluation survey.
Nottingham University Hospitals (NUH) NHS Trust provides a care pathway for patients undergoing stoma formation surgery, with the stoma care nurse specialists providing ongoing support for patients, alongside the rest of the multidisciplinary team, from the preoperative consultation through to their community follow-up. In the past, the community service offered has not always been well-received and the whole pathway has not been evaluated in depth to date. This article reports on a patient evaluation survey of the stoma care pathway at NUH. The results showed that overall patients are very complimentary of the service offered to them by the stoma care team. They highly commend the stoma care nurse specialist. Two areas for amendment in the pathway have been identified: information provision and the length of time available for the stoma care nurse specialist to spend one-to-one with each patient. The results of this survey will aid the team in implementing positive changes to the stoma care pathway.


[Study on the surgery of malignant biliary tumors (author's transl)].
The principle of the surgical therapy of malignant biliary tumor lies, as in other cancers, in the extensive removal of the tumors and lymph nodes in the regional area. Nevertheless, many problems remain unsolved with biliary carcinoma because of the anatomical structure and ensuing various symptoms depending on the degree of expansion. Seventy-two cases of biliary cancer were recorded during the past 13 years of observation in our clinic. The overall removal rate was 28%, which was extremely low as compared with other cancers. The regional removal rate was 62% in the ampulla, 25% in the gallbladder and 19% in the bile duct. In the bile duct carcinoma jaundice was most frequently seen and the degree and duration of jaundice were well correlated with the operative prognosis.


An unusual case of marble intraocular foreign body.
This report presents a case of marble intraocular foreign body that developed toxic complications during surgery. The patient is a 25 years old male who presented to the University of Benin Teaching Hospital with a history of trauma to the right eye while cutting marble. He was examined, had an ocular ultrasound scan and subsequently had an extracapsular cataract extraction. His visual acuity in the right eye was light perception. There was an entry point on the cornea, the lens was opaque, there was vitreous haemorrhage and the intraocular foreign body was localized in the posterior part of the posterior segment by ultrasound scan. He had extracapsular cataract extraction. During anterior capsulotomy, the cornea suddenly and rapidly became cloudy with a brownish tinge and the corneal epithelium started desquamating.Marble on its own may not be toxic but the other chemicals including cement, used in the processing of the marble were responsible for this delayed toxicity. Ultrasound scan is valuable in localisation of intraocular foreign bodies.


Elimination of interference from water in KBr disk FT-IR spectra of solid biomaterials by chemometrics solved with kinetic modeling.
Infrared analysis of proteins and polysaccharides by the well known KBr disk technique is notoriously frustrated and defeated by absorbed water interference in the important amide and hydroxyl regions of spectra. This interference has too often been overlooked or ignored even when the resulting distortion is critical or even fatal, as in quantitative analyses of protein secondary structure, because the water has been impossible to measure or eliminate. Therefore, a new chemometric method was devised that corrects spectra of materials in KBr disks by mathematically eliminating the water interference. A new concept termed the Beer-Lambert law absorbance ratio (R-matrix) model was augmented with water concentration ratios computed via an exponential decay kinetic model of the water absorption process in KBr, which rendered the otherwise indeterminate system of linear equations determinate and thus possible to solve in a formal analytic manner. Consequently, the heretofore baffling KBr water elimination problem is now solved once and for all. Using the new formal solution, efforts to eliminate water interference from KBr disks in research will be defeated no longer. Resulting spectra of protein were much more accurate than attenuated total reflection (ATR) spectra corrected using the well-accepted Advanced ATR Correction Algorithm.


Risk Assessment in Child Sexual Abusers Working With Children.
Child sexual abuse occurring in a child- or youth-serving institution or organization has attracted great public and scientific attention. In light of the particular personal and offense-related characteristics of men who have abused children within such an institution or organization, it is of special importance to evaluate the predictive performance of currently applied risk assessment instruments in this offender population. Therefore, the present study assessed the risk ratings and predictive performance of four risk assessment instruments and one instrument assessing protective factors concerning any, violent and sexual recidivism in child sexual abusers working with children (CSA-W) in comparison with extra-familial child sexual abusers (CSA-E) and intra-familial child sexual abusers (CSA-I). The results indicate that CSA-W mostly recidivate with a sexual offense. Although all included risk measures seem to function with CSA-W, the Static-99 seems to be the instrument that performs best in predicting sexual recidivism in CSA-W. CSA-W had the most protective factors measured with the Structured Assessment of PROtective Factors (SAPROF). While the SAPROF could not predict desistance from recidivism in CSA-W, it predicted desistance from any recidivism in all CSA. As CSA-W frequently hold many indicators for pedophilic sexual interests but only a few for antisocial tendencies, it can be suggested that CSA-W are at an increased risk for sexual recidivism and thus risk measures especially designed for sexual recidivism work best in CSA-W. Nevertheless, CSA-W also hold many protective factors; however, their impact on CSA-W is not clear yet and needs further study.


The neuronal protein Neurexin directly interacts with the Scribble-Pix complex to stimulate F-actin assembly for synaptic vesicle clustering.
Synaptic vesicles (SVs) form distinct pools at synaptic terminals, and this well-regulated separation is necessary for normal neurotransmission. However, how the SV cluster, in particular synaptic compartments, maintains normal neurotransmitter release remains a mystery. The presynaptic protein Neurexin (NRX) plays a significant role in synaptic architecture and function, and some evidence suggests that NRX is associated with neurological disorders, including autism spectrum disorders. However, the role of NRX in SV clustering is unclear. Here, using the neuromuscular junction at the 2-3 instar stages of Drosophila larvae as a model and biochemical imaging and electrophysiology techniques, we demonstrate that Drosophila NRX (DNRX) plays critical roles in regulating synaptic terminal clustering and release of SVs. We found that DNRX controls the terminal clustering and release of SVs by stimulating presynaptic F-actin. Furthermore, our results indicate that DNRX functions through the scaffold protein Scribble and the GEF protein DPix to activate the small GTPase Ras-related C3 Botulinum toxin substrate 1 (Rac1). We observed a direct interaction between the C-terminal PDZ-binding motif of DNRX and the PDZ domains of Scribble and that Scribble bridges DNRX to DPix, forming a DNRX-Scribble-DPix complex that activates Rac1 and subsequently stimulates presynaptic F-actin assembly and SV clustering. Taken together, our work provides important insights into the function of DNRX in regulating SV clustering, which could help inform further research into pathological neurexin-mediated mechanisms in neurological disorders such as autism.


Building an effective malaria vaccine pipeline to address global needs.
Despite impressive gains over the last 15 years in reducing the mortality associated with malaria, it remains a public health emergency. New interventions, such as vaccines, are needed to ensure that previous gains serve as a foundation for future progress. Vaccines have the potential to prevent severe disease and death in those most vulnerable, and to accelerate elimination and eradication by breaking the cycle of parasite transmission. The pipeline is as healthy as it has ever been, with approaches targeting different stages of the parasite lifecycle using an array of technologies. This article reviews recent progress and reviews key considerations in the quest to develop products that are aligned with the unmet medical need.


Frequency response analysis of the diaphragm in vivo.
Frequency response analysis was used to determine the dynamic response characteristics of cat diaphragm under isovolumetric conditions at functional residual capacity (FRC) and at lung volumes above and below FRC. In apneic cats, sinusoidally modulated pulse rate patterns were applied to both phrenic nerves. Modulation frequencies over the range of 0.05-4 Hz were used. Amplitude ratio vs. frequency plots obtained at FRC for intratracheal, intraesophageal, and intragastric pressures were essentially flat at low frequencies but decreased at higher frequencies. Intratracheal and intraesophageal pressure responses were altered by changes in resting lung volume while intragastric pressure was not. The amplitude ratio was decreased at lung volumes above FRC but increased at volumes below FRC. Thus, lung volume significantly affected the input-output relations between phrenic nerve input and diaphragm muscle output. In all preparations studied, significant phase lags were present throughout the entire modulation frequency range. However, in contrast to the effect of lung volume on amplitude ratio, phase lag was not dependent on changes in lung volume.


Identifying Loci Associated With Bovine Corona Virus Infection and Bovine Respiratory Disease in Dairy and Feedlot Cattle.
Bovine coronavirus (BCoV) is associated with respiratory and enteric infections in both dairy and beef cattle worldwide. It is also one of a complex of pathogens associated with bovine respiratory disease (BRD), which affects millions of cattle annually. The objectives of this study were to identify loci and heritability estimates associated with BCoV infection and BRD in dairy calves and feedlot cattle. Dairy calves from California (n = 1,938) and New Mexico (n = 647) and feedlot cattle from Colorado (n = 915) and Washington (n = 934) were tested for the presence of BCoV when classified as BRD cases or controls following the McGuirk scoring system. Two comparisons associated with BCoV were investigated: (1) cattle positive for BCoV (BCoV+) were compared to cattle negative for BCoV (BCoV-) and (2) cattle positive for BCoV and affected with BRD (BCoV+BRD+) were compared to cattle negative for BCoV and BRD (BCoV-BRD-). The Illumina BovineHD BeadChip was used for genotyping, and genome-wide association analyses (GWAA) were performed using EMMAX (efficient mixed-model association eXpedited). The GWAA for BCoV+ identified 51 loci (p < 1 × 10-5; 24 feedlot, 16 dairy, 11 combined) associated with infection with BCoV. Three loci were associated with BCoV+ across populations. Heritability estimates for BCoV+ were 0.01 for dairy, 0.11 for feedlot cattle, and 0.03 for the combined population. For BCoV+BRD+, 80 loci (p < 1 × 10-5; 26 feedlot, 25 dairy, 29 combined) were associated including 14 loci across populations. Heritability estimates for BCoV+BRD+ were 0.003 for dairy, 0.44 for feedlot cattle, and 0.07 for the combined population. Several positional candidate genes associated with BCoV and BRD in this study have been associated with other coronaviruses and respiratory infections in humans and mice. These results suggest that selection may reduce susceptibility to BCoV infection and BRD in cattle.


A Molecular Survey of Babesia Species and Detection of a New Babesia Species by DNA Related to B. venatorum from White Yaks in Tianzhu, China.
Bovine babesiosis is a tick-transmitted disease caused by different species of Babesia. The white yak is a unique yak breed that lives only in Tianzhu in the Tibetan Autonomous County, Gansu Province, in northwestern China. Previous research using the ELISA method has confirmed that the white yak could become infected with B. bigemina. The objective of this study was the molecular detection and identification of Babesia species in white yaks. A total of 409 white yak blood samples were collected from 11 areas of the Tianzhu Tibetan Autonomous County in Northwest China from April to August, 2015. The V4 hypervariable region of Babesia 18S rRNA was amplified from extracted genomic DNA using nested PCR and sequenced. The nearly full-length sequence of 18S rRNA including the V4 region from the newly discovered Babesia was amplified and sequenced with Sanger method. PCR detection and sequencing indicated that 4/409 samples were positive for B. bigemina, 3/409 samples were positive for B. bovis, and 5/409 samples were positive for B. ovata. Additionally, a new Babesia species was found in 4/409 white yaks. A unique sequence of 1,627 bp was obtained from two of the four samples. The sequence was similar to Babesia species Akita (98.5%) found in Ixodes ovatus and B. venatorum (98%) and shared a 98% identity with B. divergens and a 98.1% identity with B. odocoilei. This study provides new data about Babesia infections in white yaks in northwestern China, and a new Babesia species similar to B. venatorum was identified in white yaks for the first time.


Analysis of ACE inhibitor drugs by high performance liquid chromatography.
Seven angiotensin-converting-enzyme (ACE) inhibitors have been analyzed by HPLC. The influence of different organic modifiers and counter-ions in the eluent at different pH values have been investigated allowing the identification of the best experimental conditions for the analysis of these compounds. The sinergic effect of TEA and propanol in the eluents resulted as a critical factor to obtain satisfactory peak shapes and resolution.


Mouse Gambling Task reveals differential effects of acute sleep debt on decision-making and associated neurochemical changes.
Sleep loss is associated with sleepiness, sustained attention, and memory deficits. However, vulnerability of higher cognitive processes (i.e. decision making) to sleep debt is less understood. Therefore, a major challenge is to understand why and how higher cognitive processes are affected by sleep debt. We had established in mice correlations between individual decision-making strategies, prefrontal activity, and regional monoaminergic levels. Now, we show that acute sleep debt (ASD) disturbs decision-making processes and provokes brain regional modifications of serotonin and dopamine that could explain why ASD promotes inflexible and more risk-prone behaviors. Finally, we highlight, for the first time, that in a large group of healthy inbred mice some of them are more sensitive to ASD by showing inflexible behavior and decision-making deficits. We were also able to predict mice that would be the most vulnerable to ASD depending of their behavior before ASD exposure.


Hydrocarbon-soluble calcium hydride: a "worker-bee" in calcium chemistry.
The reactivity of the hydrocarbon-soluble calcium hydride complex [{CaH(dipp-nacnac)(thf)}(2)] (1; dipp-nacnac=CH{(CMe)(2,6-iPr(2)C(6)H(3)N)}(2)) with a large variety of substrates has been investigated. Addition of 1 to C=O and C=N functionalities gave easy access to calcium alkoxide and amide complexes. Similarly, reduction of the C[triple chemical bond]N bond in a cyanide or an isocyanide resulted in the first calcium aldimide complexes [Ca{N=C(H)R}(dipp-nacnac)] and [Ca{C(H)=NR}(dipp-nacnac)], respectively. Complexation of 1 with borane or alane Lewis acids gave the borates and alanates as contact ion pairs. In reaction with epoxides, nucleophilic ring-opening is observed as the major reaction. The high reactivity of hydrocarbon-soluble 1 with most functional groups contrasts strongly with that of insoluble CaH(2), which is essentially inert and is used as a common drying agent. Crystal structures of the following products are presented: [{Ca{OC(H)Ph(2)}(dipp-nacnac)}(2)], [{Ca{N=C(H)Ph}(dipp-nacnac)}(2)], [{Ca{C(H)=NC(Me)(2)CH(2)C(Me)(3)}(dipp-nacnac)}(2)], [{Ca{C(H)=NCy}(dipp-nacnac)}(2)], [Ca(dipp-nacnac)(thf)](+)[H(2)BC(8)H(14)](-) and [{Ca(OCy)(dipp-nacnac)}(2)]. The generally smooth and clean conversions of 1 with a variety of substrates and the stability of most intermediates against ligand exchange make 1 a valuable key precursor in the syntheses of a wide variety of beta-diketiminate calcium complexes.


Crystallization and preliminary X-ray crystallographic analysis of hydroquinone dioxygenase from Sphingomonas sp. TTNP3.
Hydroquinone dioxygenase (HQDO), a novel Fe(II) ring-fission dioxygenase from Sphingomonas sp. strain TTNP3 which oxidizes a wide range of hydroquinones to the corresponding 4-hydroxymuconic semialdehydes, has been crystallized. The enzyme is an α(2)β(2) heterotetramer constituted of two subunits of 19 and 38 kDa. Diffraction-quality crystals of HQDO were obtained using the sitting-drop vapour-diffusion method at 277 K from a solution consisting of 16% PEG 4000, 0.3 M MgCl(2), 0.1 M Tris pH 8.5. The crystals belonged to the monoclinic space group P2(1), with unit-cell parameters a = 88.4, b = 125.4, c = 90.8 Å, β = 105.3°. The asymmetric unit contained two heterotetramers, i.e. four copies of each of the two different subunits related by noncrystallographic 222 symmetry. A complete data set extending to a maximum resolution of 2.5 Å was collected at 100 K using a wavelength of 0.980 Å.


Telecom-band-integrated multimode photonic quantum memory.
Telecom-band-integrated quantum memory is an elementary building block for developing quantum networks compatible with fiber communication infrastructures. Toward such a network with large capacity, an integrated multimode photonic quantum memory at telecom band has yet been demonstrated. Here, we report a fiber-integrated multimode quantum storage of single photon at telecom band on a laser-written chip. The storage device is a fiber-pigtailed Er3+:LiNbO3 waveguide and allows a storage of up to 330 temporal modes of heralded single photon with 4-GHz-wide bandwidth at 1532 nm and a 167-fold increasing of coincidence detection rate with respect to single mode. Our memory system with all-fiber addressing is performed using telecom-band fiber-integrated and on-chip components. The results represent an important step for the future quantum networks using integrated photonics devices.


Effects of dietary calcium fructoborate supplementation on joint comfort and flexibility and serum inflammatory markers in dogs with osteoarthritis.
Our objective was to evaluate the short-term effects of calcium fructoborate (CFB) on gait, joint range of motion, serum inflammatory markers, and owner perception of pain in client-owned dogs. We used 59 osteoarthritic dogs with impairment, with dogs being randomly assigned to 4 treatments: placebo (60 mg fructose; = 15), low dose (69 mg CFB; = 14), high dose (127 mg CFB; = 14), or combination (69 mg CFB, 500 mg glucosamine hydrochloride and 200 mg chondroitin sulfate; = 16). Dogs up to 22.9 kg received 1 capsule/d, while dogs weighing 23 to 50 kg received 2 capsules/d. A physical examination, radiographs, goniometry measurements, gait analysis, blood sample collection, and a canine brief pain inventory questionnaire were performed on d 0 and 28. Change from baseline values were statistically analyzed among groups. After 28 d, dogs fed the low and high doses had an improved ( < 0.05) ability to rise from a lying position compared to placebo. Dogs fed the high dose also had a greater ( = 0.05) increase in soluble receptor for advanced glycation end products concentration than dogs fed the placebo. Sub-analysis of only large dogs (> 23 kg) showed that dogs fed the low dose had decreased ( < 0.05) pain severity score and pain at its worst compared to dogs fed the placebo. Large dogs fed the low dose also were shown to improve ( < 0.05) in their ability to rise from a lying position compared to dogs fed the placebo. Overall, CFB supplementation was well-tolerated and may aid in mitigating joint discomfort in dogs.


Lack of Glycogenin Causes Glycogen Accumulation and Muscle Function Impairment.
Glycogenin is considered essential for glycogen synthesis, as it acts as a primer for the initiation of the polysaccharide chain. Against expectations, glycogenin-deficient mice (Gyg KO) accumulate high amounts of glycogen in striated muscle. Furthermore, this glycogen contains no covalently bound protein, thereby demonstrating that a protein primer is not strictly necessary for the synthesis of the polysaccharide in vivo. Strikingly, in spite of the higher glycogen content, Gyg KO mice showed lower resting energy expenditure and less resistance than control animals when subjected to endurance exercise. These observations can be attributed to a switch of oxidative myofibers toward glycolytic metabolism. Mice overexpressing glycogen synthase in the muscle showed similar alterations, thus indicating that this switch is caused by the excess of glycogen. These results may explain the muscular defects of GSD XV patients, who lack glycogenin-1 and show high glycogen accumulation in muscle.


Medication-related osteonecrosis of the jaw after long-term bisphosphonate treatment in a cat.
A 12-year-old, neutered female, domestic medium hair cat was evaluated for a nonhealing, oral mucosal ulceration. The cat had a history of idiopathic hypercalcemia that had been treated with a bisphosphonate for 41 months. Oral examination identified exposed maxillary bone adjacent to a previous extraction site. Histopathology of the exposed bone and associated mucosa was most consistent with medication-related osteonecrosis of the jaw. Treatment involved both medical and surgical interventions. Oral mucosal healing occurred after 6 months of treatment.


The impact of climate change on infectious disease transmission: perceptions of CDC health professionals in Shanxi Province, China.
There have been increasing concerns about the challenge of emerging and re-emerging infectious diseases due to climate change, especially in developing countries including China. Health professionals play a significant role in the battle to control and prevent infectious diseases. This study therefore aims to investigate the perceptions and attitudes of health professionals at the Centers for Disease Control and Prevention (CDC) in different levels in China, and to consider adaptation measures to deal with the challenge of climate change. In 2013, a cross-sectional questionnaire survey was undertaken among 314 staff in CDCs in Shanxi Province, China, whose routine work involves disease control and prevention. Data were analyzed using descriptive methods and logistic regression. A majority of the CDC staff were aware of the health risks from climate change, especially its impacts on infectious disease transmission in their jurisdictions, and believed climate change might bring about both temporal and spatial change in transmission patterns. It was thought that adaptation measures should be established including: strengthening/improving currently existing disease surveillance systems and vector monitoring; building CDC capacity in terms of infrastructure and in-house health professional training; development and refinement of relevant legislation, policies and guidelines; better coordination among various government departments; the involvement of the community in infectious disease interventions; and collaborative research with other institutions. This study provides a snapshot of the understanding of CDC staff regarding climate change risks relevant to infectious diseases and adaptation in China. Results may help inform future efforts to develop adaptation measures to minimize infectious disease risks due to climate change.


Pretreatment with and ongoing use of omega-3 fatty acid ethyl esters reduce the slow-flow phenomenon and prevent in-stent restenosis in patients undergoing carotid artery stenting.
Carotid artery stenting (CAS) is a less invasive alternative to carotid endarterectomy, but it is essential to prevent thromboembolic complications during CAS and to suppress in-stent restenosis (ISR) after CAS because of the relatively high risk of periprocedural and follow-up stroke events. Clinical trials have demonstrated the strong relationship of carotid plaque vulnerability with the subsequent risk of ipsilateral ischemic stroke and thromboembolic complications during CAS. Recent studies demonstrated that both low eicosapentaenoic acid (EPA) and low docosahexaenoic acid (DHA) levels were significantly associated with lipid-rich coronary and carotid plaques, but little is known about the effect of administration of omega-3 fatty acids (O-3FAs) containing EPA and DHA before and after CAS for stabilizing carotid plaque, preventing thromboembolic complications, and suppressing ISR. In this study, the efficacy of pretreatment with and ongoing daily use of O-3FA in addition to statin treatment was evaluated in patients undergoing CAS. This study was a nonrandomized prospective trial with retrospective analysis of historical control data. From 2012 to 2015, there were 100 consecutive patients with hyperlipidemia undergoing CAS for carotid artery stenosis who were divided into two groups. Between 2012 and 2013 (control period), 47 patients were treated with standard statin therapy. Between 2014 and 2015 (O-3FA period), patients were treated with statin therapy and add-on oral O-3FA ethyl esters containing 750 mg/d DHA and 1860 mg/d EPA from 4 weeks before CAS, followed by ongoing daily use for at least 12 months. In all patients, the plaque morphology by virtual histology intravascular ultrasound, the incidence of new ipsilateral ischemic lesions on the day after CAS, the slow-flow phenomenon during CAS, and ISR within 12 months after CAS were compared between the periods. The slow-flow phenomenon during CAS with filter-type embolic protection devices decreased in the O-3FA period (1 of 53 patients [2%]) compared with the control period (7 of 47 patients [15%]; P = .02). Furthermore, ISR for 12 months after CAS was significantly decreased in the O-3FA period (1 of 53 patients [2%]) compared with the control period (10 of 47 patients [21%]; P = .01). On virtual histology intravascular ultrasound analysis, the fibrofatty area was significantly smaller and the fibrous area was significantly greater in the O-3FA period. On multivariate logistic regression analysis, a low EPA/arachidonic acid ratio and a symptomatic lesion were the factors related to vulnerable plaque (P = .01 [odds ratio, 5.24; 95% confidence interval, 1.65-16.63] and P = .01 [odds ratio, 11.72; 95% confidence interval, 2.93-46.86], respectively). Pretreatment with O-3FA reduces the slow-flow phenomenon generated by plaque vulnerability during CAS, and on-going daily use of O-3FA suppresses ISR after CAS.


Ability of the Listeria monocytogenes strain Scott A to cause systemic infection in mice infected by the intragastric route.
Listeriosis is an important food-borne disease that causes high rates of morbidity and mortality. For reasons that are not clear, most large outbreaks of human listeriosis involve Listeria monocytogenes serotype 4b. Relatively little is known about the pathogenesis of listeriosis following gastrointestinal exposure to food-borne disease isolates of L. monocytogenes. In the present study, we investigated the pathogenesis of systemic infection by the food-borne isolate Scott A in an intragastric (i.g.) mouse challenge model. We found that the severity of infection with L. monocytogenes Scott A was increased in mice made neutropenic by administration of monoclonal antibody RB6-8C5. This observation was similar to a previous report on a study with the laboratory strain L. monocytogenes EGD. Prior administration of sodium bicarbonate did not enhance the virulence of L. monocytogenes strain Scott A for i.g. inoculated mice. Following i.g. inoculation of mice, two serotype 4b strains of L. monocytogenes (Scott A and 101M) achieved a greater bacterial burden in the spleen and liver and elicited more severe histopathological damage to those organs than did a serotype 1/2a strain (EGD) and a serotype 1/2b stain (CM). Of the four strains tested, only strain CM exhibited poor survival in synthetic gastric fluid in vitro. The other three strains exhibited similar patterns of survival at pHs of greater than 5 and relatively rapid (<30 min) loss of viability at pHs of less than 5.0. Growth of L. monocytogenes Scott A at temperatures of 12.5 to 37 degrees C did not affect its ability to cause systemic infection in i.g. inoculated mice. These observations suggest that the serotype 4b L. monocytogenes strains Scott A and 101M possess one or more virulence determinants that make them better able to cause systemic infection following inoculation via the g.i. tract than do the serotype 1/2 strains EGD and CM.


Effect of Different Surface Treatments on Porcelain-Resin Bond Strength.
The aim of this study was to evaluate the effects of various surface treatments on the surface structure and shear bond strength (SBS) of different ceramics. A total of 192 disk-shaped cores were prepared using two all-ceramic systems, of which 168 were submitted to SBS tests, and 24 were investigated by scanning electron microscopy (SEM) and atomic force microscopy (AFM). The ceramics used were IPS Empress e.max (EX) lithium glass-ceramic and Vita In-Ceram Zirconia glass-infiltrated zirconia (ICZ). The specimens were randomly divided into seven groups (n = 12) on the basis of the surface treatment used: control; SB-sandblasting with 50 μm Al2 O3 particles; CJ-chairside silica coating with 30 μm SiO2 particles and silanization (Clearfil Porcelain Bond Activator); HF-etching in 5% hydrofluoric acid and silanization; ER-etching with an Er:YAG laser (10 W); ND-Nd:YAG laser etching (0.8 W); and FS-etching with a femtosecond laser (860 mW). A luting cement (Clearfil Esthetic) was photopolymerized on each treated ceramic disk. After subjecting the specimens to thermocycling (1000 cycles, 5°C to 55°C), SBS tests were performed using a universal testing machine. The data were analyzed with two-way ANOVA and Tukey's tests using a significance limit of 5%. Among the EX ceramics, the CJ (29.10 MPa) and HF (26.07 MPa) specimens had statistically higher SBS values. For the ICZ ceramics, the highest value (28.08 MPa) was obtained for the CJ specimens. Silanization after coating with silica improves the bond strengths of both EX and ICZ specimens, while HF etching is favorable only for the EX specimens.


Indirect role of T cells in development of polioencephalitis and encephalomyelitis induced by encephalomyocarditis virus.
Infection of female BALB/c mice with encephalomyocarditis virus results in the development of a paralytic syndrome in 7 to 10 days postinoculation. Previous studies had suggested the involvement of an immune component in the development of central nervous system pathology. We have examined the effects of T-cell depletion on the development of polioencephalitis (neuronal necrosis and inflammation of the brain and brain stem) and the relative contribution of the CD4+ and CD8+ subsets following the establishment of viremia. We show that monoclonal antibody depletion of T cells is effective in the reduction of polioencephalitis when given prior to viral inoculation. However, administration of the antibodies 12 h or more after viral inoculation failed to alter the development of polioencephalitis or encephalomyelitis. We conclude that T cells are involved in the development of central nervous system disease during the initial stages of infection but are not responsible for the later progression of disease.


What is new in pain modification in osteoarthritis?
There is a big need for the development of novel therapies for the safe management of chronic pain associated with OA. Here we reviewed PubMed (2015 onward) and ClinicalTrials.gov for ongoing and recently completed trials where pain in OA is the primary outcome measure. Three broad categories were identified: biological therapies, small molecules and cryoneurolysis. The most promising new strategy is blockade of nerve growth factor with antibodies. Two anti-nerve growth factor antibodies, tanuzemab and fasinumab, are in active development after the 2010 hold on trials was lifted in 2015. In addition, several active clinical trials are testing distinct mechanism-based interventions, including cytokine inhibition, selective μ, δ or κ opioid receptor agonists, zoledronate and intra-articular capsaicin. In addition to pharmacological approaches, cryoneurolytic strategies that directly target peripheral nerves may play a role in OA pain management, but efficacy profiles and long-term effects of such treatments need more study. Clearly, the therapeutic landscape for OA pain is rapidly expanding. Since symptomatic OA is a heterogeneous disease, the challenge will be to identify patients that will benefit the most from specific approaches.


Two-Dimensional Gel Electrophoresis.
Improved resolution of serum protein mixtures is effected by electrophoresis, first in a 5 percent acrylamide gel following which a strip of the resolved pattern is embedded in 8 percent gel and subjected to a second electrophoresis separation at right angles to the first. Lactic dehydrogenase enzymes appear as small rectangular spots lying on an oblique straight line passing through the point of initial application.


Choosing to eat your way toward healthy aging.
As individuals age, they face many choices to promote healthy aging. One important consideration relates to nutritional choices. It is easy to become overwhelmed with the information in the media that suggests the superiority of some new diet or breakthrough dietary supplement. This article reminds us to focus on holistic approaches to nutrition on the premise that balance is paramount in our lives. As nurses, we need to guide others to make thoughtful choices for healthy aging.


Association between cancer stem cell gene expression signatures and prognosis in head and neck squamous cell carcinoma.
Various cancer stem cell (CSC) biomarkers and the genes encoding them in head and neck squamous cell carcinoma (HNSCC) have been identified and evaluated. However, the validity of these factors in the prognosis of HNSCC has been questioned and remains unclear. In this study, we examined the clinical significance of CSC biomarker genes in HNSCC, using five publicly available HNSCC cohorts. To predict the prognosis of patients with HNSCC, we developed and validated the expression signatures of CSC biomarker genes whose mRNA expression levels correlated with at least one of the four CSC genes (CD44, MET, ALDH1A1, and BMI1). Patients in The Cancer Genome Atlas (TCGA) HNSCC cohort were classified into CSC gene expression-associated high-risk (CSC-HR; n = 285) and CSC gene expression-associated low-risk (CSC-LR; n = 281) subgroups. The 5-year overall survival and recurrence-free survival rates were significantly lower in the CSC-HR subgroup than in the CSC-LR subgroup (p = 0.04 and 0.02, respectively). The clinical significance of the CSC gene expression signature was validated using four independent cohorts. Analysis using Cox proportional hazards models showed that the CSC gene expression signature was an independent prognostic factor of non-oropharyngeal HNSCC which mostly indicates HPV (-) status. Furthermore, the CSC gene expression signature was associated with the prognosis of HNSCC patients who received radiotherapy. The CSC gene expression signature is associated with the prognosis of HNSCC and may help in personalized treatments for patients with HNSCC, especially in cases with HPV (-) status who were classified in more detail.


Health professionals' attitude toward euthanasia: a cross-sectional study from Turkey.
There is an ongoing debate on the definition of euthanasia and attempts to change laws about euthanasia and its practice in many countries. It is the medical doctors and the other health professionals who will elucidate the issue. Therefore, we performed this study to reveal attitude of Turkish health professionals toward euthanasia. This is an observational and cross-sectional study and data were collected with a questionnaire. The questionnaire was distributed to 545 health professionals in Samsun, a city in the Black Sea Region in Turkey. Data were analyzed with SPSS package programs. Of all health professionals included in the study, 43.5% were medical doctors and 45.5% auxiliary health professionals. Of all participants, 33.6% did not object to euthanasia and 7.9% were asked to perform euthanasia. Eighty point seven percent of the participants noted that euthanasia could be abused even if a euthanasia law were passed. It can be concluded that the health professionals should have a chance to discuss euthanasia and that their attitude toward and their expectations and worries about euthanasia should be taken into account when a euthanasia law is drafted.


Two bifunctional enzymes from the marine protist Thraustochytrium roseum: biochemical characterization of wax ester synthase/acyl-CoA:diacylglycerol acyltransferase activity catalyzing wax ester and triacylglycerol synthesis.
Triacylglycerols (TAGs) and wax esters (WEs) are important neutral lipids which serve as energy reservoir in some plants and microorganisms. In recent years, these biologically produced neutral lipids have been regarded as potential alternative energy sources for biofuel production because of the increased interest on developing renewable and environmentally benign alternatives for fossil fuels. In bacteria, the final step in TAG and WE biosynthetic pathway is catalyzed by wax ester synthase/acyl coenzyme A (acyl-CoA):diacylglycerol acyltransferase (WS/DGAT). This bifunctional WS/DGAT enzyme is also a key enzyme in biotechnological production of liquid WE via engineering of plants and microorganisms. To date, knowledge about this class of biologically and biotechnologically important enzymes is mainly from biochemical characterization of WS/DGATs from Arabidopsis, jojoba and some bacteria that can synthesize both TAGs and WEs intracellularly, whereas little is known about WS/DGATs from eukaryotic microorganisms. Here, we report the identification and characterization of two bifunctional WS/DGAT enzymes (designated TrWSD4 and TrWSD5) from the marine protist Thraustochytrium roseum. Both TrWSD4 and TrWSD5 comprise a WS-like acyl-CoA acyltransferase domain and the recombinant proteins purified from Escherichia coli Rosetta (DE3) have substantial WS and lower DGAT activity. They exhibit WS activity towards various-chain-length saturated and polyunsaturated acyl-CoAs and fatty alcohols ranging from C10 to C18. TrWSD4 displays WS activity with the lowest Km value of 0.14 μM and the highest kcat/Km value of 1.46 × 105 M-1 s-1 for lauroyl-CoA (C12:0) in the presence of 100 μM hexadecanol, while TrWSD5 exhibits WS activity with the lowest Km value of 0.96 μM and the highest kcat/Km value of 9.83 × 104 M-1 s-1 for decanoyl-CoA (C10:0) under the same reaction condition. Both WS/DGAT enzymes have the highest WS activity at 37 and 47 °C, and WS activity was greatly decreased when temperature exceeds 47 °C. TrWSD4 and TrWSD5 are insensitive to ionic strength and reduced WS activity was observed when salt concentration exceeded 800 mM. The potential of T. roseum WS/DGATs to establish novel process for biotechnological production of WEs was demonstrated by heterologous expression in recombinant yeast. Expression of either TrWSD4 or TrWSD5 in Saccharomyces cerevisiae quadruple mutant H1246, which is devoid of storage lipids, resulted in the accumulation of WEs, but not any detectable TAGs, indicating a predominant WS activity in yeast. This study demonstrates both in vitro WS and DGAT activity of two T. roseum WS/DGATs, which were characterized as unspecific acyltransferases accepting a broad range of acyl-CoAs and fatty alcohols as substrates for WS activity but displaying substrate preference for medium-chain acyl-CoAs. In vivo characterization shows that these two WS/DGATs predominantly function as wax synthase and presents the feasibility for production of WEs by heterologous hosts.


Chaos Structure and Contagion Behavior between COVID-19, and the Returns of Prices of Precious Metals and Oil: MS-GARCH-MLP Copula.
This study was developed for two purposes. The first one is detection of the existence of chaotic structure and uncertainty behaviour of the total number of people infected with the COVID-19 outbreak, and the returns of precious metals and oil by using the Lyapunov exponent, Kolmogrov and Shannon entropy tests. The additional aim was to analyze the co-movement and contagion behavior among COVID-19 infected persons, returns of precious metals and oil for the period of between December 30, 2019 and October 26, 2020 by MSGARCH-copula and MSGARCH-multi-layer perceptron methods. Confirmation of the results was provided by using the Diebold-Mariano (DM) tests and Wilcoxon signed rank (WS) tests. Accordingly, the empirical findings of this paper are as follows: The presence of chaotic structure and uncertainty behavior in the variables were determined by Lyapunov exponent, Kolmogorov and Shannon entropy tests. Additionally, co-movement and contagion behavior among the analyzed variables was established by the MS-GARCH-MLP copula method. Following, in the context of the results of confirmation, it has been determined that MSGARCH-MLPst has the best prediction performance under all criteria compared to MSGARCHst, or GARCHst. As a further result, COVID-19 had considerable effects on the returns of precious metals and oil prices, and there was co-movement and contagion behavior between COVID-19 and the returns of precious metals and oil.


Community health workers in Ghana: the need for greater policy attention.
From the 1970s to the 1990s, the WHO, United Nations and other agencies mooted the idea of formally training and recognising community health workers (CHWs) to complement efforts to improve primary healthcare delivery in low and middle income countries. Recently, CHWs have been recognised as important players in the achievement of the health-related Millennium Development Goals (MDGs). Despite this recognition, little understanding exists in Ghana about the activities of CHWs: who they are; how they are recruited; what they do; level of health policy support; contribution to healthcare delivery and the challenges they face. Based on a rapid scoping review of the existing literature, and our experience working in Ghana, this paper reflects on the role of CHWs in healthcare delivery in Ghana. We argue that CHWs have played critical roles in improving health service delivery and outcomes, including guinea worm eradication, expanded immunisation coverage, maternal and child health, and HIV/AIDS treatment and management. However, these achievements notwithstanding, CHWs face challenges which prevent them from being optimally productive, including capacity problems, neglect by the healthcare system, high attrition rates and inadequate supervision. Policymakers in Ghana therefore need to give increased attention to CHWs, provide remuneration for their activities, create career opportunities and other means of motivations to boost their productivity and sustain gains associated with their activities.


Urinary bile casts in bile cast nephropathy secondary to severe falciparum malaria.
Severe cholestatic jaundice may complicate with bile cast nephropathy (BCN) causing severe acute kidney injury (AKI). In this study, we investigate BCN in severe falciparum malaria complicated with jaundice and AKI. This prospective study was conducted in a tertiary health care institution with high prevalence of malaria. A cohort of 110 patients with falciparum malaria complicated with cerebral malaria, jaundice and AKI were enrolled. Species diagnosis was made from peripheral blood smear or rapid diagnostic test. Severe malaria was diagnosed from WHO criteria. BCN was diagnosed with the detection of bile casts in urine or in biopsy. The recovery pattern and outcome with and without BCN was assessed. Out of 110 patients, 20 (18.2%) patients had BCN and 15 (13.6%) patients had hepato-renal syndrome. Patients with BCN had high conjugated bilirubin (26.5 ± 4.1 mg/dL), urea (75.9 ± 10.3 mg/dL) and creatinine (7.2 ± 0.8 mg/dL), longer duration of illness (6.4 ± 1.1 days), higher mortality (25.0%) and prolonged recovery time of hepatic (9.6 ± 2.4 days) and renal dysfunction (15.1 ± 6.5 days) compared with patients without BCN. Prolonged duration of illness and increased bilirubin cause BCN among patients with severe falciparum malaria with jaundice and AKI, which is associated with high mortality and morbidity.


Role of inflammasomes in intestinal inflammation and Crohn's disease.
: Inflammasomes are multiprotein complexes that process procytokines into mature forms of interleukin 1β and interleukin 18 and induce pyroptotic cell death. Evidence linking NLRP3, NLRC4, and NLRP6 inflammasomes to intestinal inflammation is reviewed to provide a basis to understand how the innate immune system discriminates pathogenic bacteria from commensal bacteria and shapes microbial ecology. Inflammasomes have a direct and important role limiting colitis by directing effective immune responses against pathogenic bacterial infections in the intestine. Chronic granulomatous disease is presented to reveal a contrasting proinflammatory effect of inflammasomes. This pathogenic effect is unmasked in a state of immunodeficiency where bacterial growth is poorly controlled increasing inflammasome activity. The role of inflammasomes in inflammation associated with Crohn's disease and ulcerative colitis is discussed. Finally, mechanistic studies linking genetic polymorphisms in ATG16L and NOD2 to inflammasome activation provide a basis for new hypotheses to explain how genetic polymorphism associated with Crohn's disease modulate intestinal inflammation. A deeper understanding of the role of inflammasomes in intestinal inflammation is expected to identify new ways of treating inflammatory bowel disease.