Single-step co-integration of multiple expressible heterologous genes into the ribosomal DNA of the methylotrophic yeast Hansenula polymorpha.
We have investigated the methylotrophic yeast Hansenula polymorpha as a host for the co-integration and expression of multiple heterologous genes using an rDNA integration approach. The ribosomal DNA (rDNA) of H. polymorpha was found to consist of a single rDNA cluster of about 50-60 repeats of an 8-kb unit located on chromosome II. A 2.4-kb segment of H. polymorpha rDNA encompassing parts of the 25S, the complete 5S and the non-transcribed spacer region between 25S and 18S rDNA was isolated and inserted into conventional integrative H. polymorpha plasmids harboring the Saccharomyces- cerevisiae-derived URA3 gene for selection. These rDNA plasmids integrated homologously into the rDNA repeats of a H. polymorpha (odc1) host as several independent clusters. Anticipating that this mode of multiple-cluster integration could be used for the simultaneous integration of several distinct rDNA plasmids, the host strain was co-transformed with a mixture of up to three different plasmids, all bearing the same URA3 selection marker. Transformations indeed resulted in mitotically stable strains harboring one, two, or all three plasmids integrated into the rDNA. The overall copy number of the plasmids integrated did not exceed the number of rDNA repeats present in the untransformed host strain, irrespective of the number of different plasmids involved. Strains harboring different plasmids co-expressed the introduced genes, resulting in functional proteins. Thus, this approach provides a new and attractive tool for the rapid generation of recombinant strains that simultaneously co-produce several proteins in desired stoichiometric ratios.


Oseltamivir phosphate monotherapy ablates tumor neovascularization, growth, and metastasis in mouse model of human triple-negative breast adenocarcinoma.
Triple-negative breast cancers (TNBCs) lack the estrogen, progesterone, and epidermal growth factor (EGF) receptor-2 (HER2/neu) receptors. Patients with TNBC have typical high grading, more frequent relapses, and exhibit poorer outcomes or prognosis compared with the other subtypes of breast cancers. Currently, there are no targeted therapies that are effective for TNBC. Preclinical antitumor activity of oseltamivir phosphate (OP) therapy was investigated to identify its role in tumor neovascularization, growth, invasiveness, and long-term survival in a mouse model of human TNBC. Live cell sialidase, water soluble tetrazolium, WST-1 cell viability, and immunohistochemistry assays were used to evaluate sialidase activity, cell survival, and the expression levels of tumor E-cadherin, N-cadherin, and host endothelial CD31+/PECAM-1 cells in archived paraffin-embedded TNBC MDA-MB-231 tumors grown in RAGxCγ double mutant mice. OP, anti-Neu1 antibodies, and matrix metalloproteinase-9-specific inhibitor blocked Neu1 activity associated with EGF-stimulated TNBC MDA-MB-231 cells. OP treatment of MDA-MB-231 and MCF-7 cells and their long-term tamoxifen-resistant clones reproducibly and dose-dependently reduced the sialidase activity associated with EGF-stimulated live cells and the cell viability after 72 hours of incubation. Combination of 1 μM cisplatin, 5-FU, paclitaxel, gemcitabine, or tamoxifen with OP dosages ≥300 μg/mL significantly reduced cell viability at 24, 48, and 72 hours when compared to the chemodrug alone. Heterotopic xenografts of MDA-MB-231 tumors developed robust and bloody tumor vascularization in RAG2xCγ double mutant mice. OP treatment at 30 mg/kg daily intraperitoneally reduced tumor vascularization and growth rate as well as significantly reduced tumor weight and spread to the lungs compared with the untreated cohorts. OP treatment at 50 mg/kg completely ablated tumor vascularization, tumor growth and spread to the lungs, with significant long-term survival at day 180 postimplantation, tumor shrinking, and no relapses after 56 days off-drug. OP 30 mg/kg cohort tumors expressed significantly reduced levels of human N-cadherins and host CD31+ endothelial cells with concomitant significant expression of E-cadherins compared to the untreated cohorts. OP monotherapy may be the effective treatment therapy for TNBC.


Effect of short-term maximal voluntary co-contraction training on neuromuscular function.
The present study aimed to examine the effect of short-term training utilizing voluntary co-contraction with maximal efforts. 23 healthy young men (training group: TG, n = 13; control group: CG, n = 10) participated in this study. TG conducted a 4-week training program (3 days/week), which consisted of 4 s simultaneous maximal voluntary contractions of elbow flexors and extensors at 90° of the elbow joint, followed by 4 s muscle relaxation (10 repetitions/set, 5 sets/day). Before and after the intervention, maximal voluntary isometric and isokinetic torques and the muscle thicknesses of the elbow flexors and extensors were determined. The electromyograms (EMGs) of the 2 muscle groups during isometric maximal voluntary contraction (MVC) were also recorded. After the intervention, CG did not show any significant changes in all measured variables. In TG, significant increases were found in the agonist EMG activities during MVC, and maximal isometric and isokinetic torques of the elbow flexors and extensors, without significant changes in the muscle thicknesses and involuntary coactivation levels during MVC. The current results indicate that the training mode with maximal voluntary co-contraction is effective for improving the force-generating capabilities of the exercising muscles, without any increases in the level of involuntary coactivation during MVC.


The behaviour of human oral squamous cell carcinoma in cell culture.
This study examined the initial behaviour of 48 human oral squamous cell carcinomas (SCC) in cell culture. The early outcome of these cultures (contamination, absence of cell growth, epithelial cell senescence/fibroblast overgrowth, extended keratinocyte growth) did not reflect the clinical characteristics of the tumours of origin. Four new human oral SCC cell lines were characterized more extensively. Each cell line was immortal, 3T3-independent, and expressed low degrees of anchorage independence (CFE less than 4 per cent). Two of the four cell lines were tumorigenic in athymic mice. All of the cell lines expressed keratin intermediate filaments and two showed weak co-expression of vimentin. A wide range of keratins were expressed by the tumour xenografts; cornified keratins (K1, K10) were only expressed in the absence of K19 and vimentin, and vice versa. The nuclear:cytoplasmic ratio and the degree of serum independence correlated with each other and with the STNMP clinical grading of the tumours of origin.


[Testing basophil activation by flow cytometry in the diagnosis of allergy to hymenopteran venom].
Diagnosis of allergy to hymenoptera venom must be precise and depends on indisputable bio-clinical criteria, because of specific immunotherapy for indications such as systemic and/or anaphylactic reactions. Until nowadays, diagnosis was by specific IgE, histamine release and skin tests, most often done for the venoms of wasp, honey bee and hornet at the same time since, in 7-8 cases in 10 the patients had not identified the responsible insect. Basophil activation test (TAB) by Flux cytometry and measurement of leukotriene C4 (LTC4) are new techniques of great reliability. The work shows the correlations between the different immunobiological parameters by reference to TAB by CAF and measurement of LTC4. When the overall results for mixed venoms or those for single venoms are considered, the correlations between TAB, LTC4 and the other parameters are highly significant. It can now be considered objectively that TAB by CTF and measurement of LCT4 are the two highest-performing techniques for diagnosis of hymenoptera venom allergy and so validates them.


Theory and modeling of self-organization and propagation of filamentary plasma arrays in microwave breakdown at atmospheric pressure.
High power microwave breakdown at atmospheric pressure leads to the formation of filamentary plasma arrays that propagate toward the source. A two-dimensional model coupling Maxwell equations with plasma fluid equations is used to describe the formation of patterns under conditions similar to recent experiments and for a wave electric field perpendicular to the simulation domain or in the simulation domain. The calculated patterns are in excellent qualitative agreement with the experiments, with good quantitative agreement of the propagation speed of the filaments. The propagation of the plasma filaments is due to the combination of diffusion and ionization. Emphasis is put on the fact that free electron diffusion (and not ambipolar diffusion) associated with ionization is responsible for the propagation of the front.


Vibrionaceae as a possible source of Qnr-like quinolone resistance determinants.
To gain insight into the functionality of qnr-like genes of several bacterial species of Vibrionaceae that may encode quinolone resistance determinants. A PCR-based strategy was used to obtain qnr-like genes of reference strains of Vibrio vulnificus CIP103196, Vibrio parahaemolyticus CIP71.2 and Photobacterium profundum CIP106289 that were sequenced, cloned and expressed in Escherichia coli. MICs of quinolones were determined for these reference strains and an E. coli reference strain harbouring recombinant plasmids. The Qnr-like proteins of these Vibrionaceae bacterial species shared 40-67% amino acid identity with the plasmid-mediated quinolone resistance determinants QnrA, QnrB and QnrS with a series of highly conserved residues. Once cloned in E. coli they conferred reduced susceptibility to quinolones. This work provides further evidence that water-borne Vibrionaceae may constitute a reservoir for Qnr-like quinolone resistance determinants.


Assessment of genotoxicity of inorganic mercury in rats in vivo using both chromosomal aberration and comet assays.
The major objective of the present investigation was to assess the genotoxic effects of mercuric chloride (HgCl2), an inorganic mercury (Hg), in rats (Rattus norvegicus) using two different genetic endpoints, namely, chromosomal aberration (CA) and comet assays following both short-term (acute) and long-term (chronic) exposures. The study showed that the acute exposures to HgCl2 at 2 and 5 mg/kg body weight (b.w.) induced nonsignificant effects. HgCl2 at 10 and 12 mg/kg b.w. was significantly toxic and is exhibited by the induction of different types of CAs like chromatid breaks, chromosomal breaks, clumps and damaged cells and types of comets. HgCl2 at 15 mg/kg b.w. was found to be highly toxic, as mitostatic condition of cells were observed in CA assay. Chronic exposure to the lowest dose (2 mg/kg b.w.) of HgCl2 for 15 consecutive days produced a significant genotoxicity. Although Hg was found to induce both DNA strand breakage and chromosomal breaks in a dose-dependent manner, the results of the present investigation showed that the combination of comet and CA assays provided a better choice for assessing the genotoxicity of inorganicHg.


Radial oxygen loss by the cushion plant Eriocaulon schimperi prevents methane emissions from an East-African mountain mire.
Groundwater-fed fens are known sources of methane (CH4 ) emissions to the atmosphere, and these are known to be mediated by the vegetation. In a fen located in the Bale Mountains, Ethiopia, we assessed the effects of a cushion plant (Eriocaulon schimperi) and a sedge (Carex monostachya) on rhizosphere biogeochemistry. Methane and CO2 concentrations and pH were measured in pore-water at different depths in the profile. Redox potentials and NaCl-extractable element concentrations were analysed in soil samples from sites dominated by either E. schimperii or C. monostachya. Nutrient and element concentration were analysed in plant tissues. At Carex-dominated sites, CH4 concentrations increased from 70 μmol·l-1 at a depth of 10 cm to 130 μmol·l-1 at a depth of 100 cm. CH4 concentrations at Eriocaulon-dominated sites were almost zero (<1 μmol·l-1 ) to a depth of 100 cm. Simultaneously, soil redox potentials and CO2 concentrations were higher at Eriocaulon-dominated sites, indicating a low potential for CH4 production and a high potential for CH4 oxidation. Eriocaulon schimperi displayed a root investment strategy to cope with the harsh environment, similar to the cushion plant Astelia pumila in Patagonian bogs. This strategy is characterised by high root/shoot ratios, high root porosity and density under high redox conditions. Both cushion plant species create an aerobic rhizosphere through radial oxygen loss from deep roots, which strongly reduce CH4 fluxes to the atmosphere.


Trace heavy metals associated with crude oil: a case study of Ebocha-8 oil-spill-polluted site in Niger Delta, Nigeria.
The Ebocha-8 oil-spill-polluted site was identified as study site according to field reconnaissance surveys of oil-polluted sites in Obiobi/Obrikom prospect area of Niger Delta. A sampling area was delimited at each of the oil-spill-affected sites by the grid technique, and soils were collected at surface (0-15 cm) and subsurface (15-30 cm) depths from three replicate quadrants. A geographically similar, unaffected area, located in 50-m distance from each contaminated site, was chosen as control site. Trace heavy metals, Ni, V, Cu, Cd, and Pb, which are normal constituents of crude oil, were determined in the soils by atomic-absorption spectrophotometry after pre-extraction of cations with dithionite-citrate carbonate. Ni varied from 0.53 to 18.00 mg/kg, Cu from 0.15 to 0.30 mg/kg, and Pb from 0.32 to 0.80 mg/kg in surface and subsurface soils, respectively; Cd and V were less than 0.20 mg/kg in all sampled plots. Ni, Cu, and Pb were more enhanced (P<0.05) in the oil-spill-polluted soils, especially at surface depth, and this may be attributed to the fact that metal profiles in polluted soils penetrate a little below the 10-cm region, even after many years, thereby making the metal concentration in surface soils usually higher. Whilst the Ebocha-8 oil spillage may be responsible, at least in part, for the enhanced concentrations and paucity of variations in the metal concentrations of sampled plots, other factors such as the physicochemical characteristics of soils (e.g., soil pH and organic-matter content) and relative mobility of these metals, as well as the intense rainfall and flooding that preceded the period of sampling, may also have contributed in part. Enhanced levels of these micronutrients in the soils may result in enhanced absorption by plants, which may bring about possible bioaccumulation by such plants and the animals that depend on them for survival, and all of these may lead to toxic reactions along the food chain.


Intracorneal Hematoma Showing Clinical and Dermoscopic Features of Acral Lentiginous Melanoma.
Intra- and subcorneal hematoma, a skin alteration seen palmar and plantar after trauma or physical exercise, can be challenging to distinguish from in situ or invasive acral lentiginous melanoma. Thus, careful examination including dermoscopic and histologic assessment may be necessary to make the correct diagnosis. We here present a case of a 67-year-old healthy female patient who presented with a pigmented plantar skin alteration. Differential diagnoses included benign skin lesions, for example, hematoma or melanocytic nevus, and also acral lentiginous melanoma or melanoma in situ. Since clinical and dermoscopic examinations did not rule out a malignant skin lesion, surgical excision was performed and confirmed an intracorneal hematoma. In summary, without adequate physical trigger, it may be clinically and dermoscopically challenging to make the correct diagnosis in pigmented palmar and plantar skin alterations. Thus, biopsy or surgical excision of the skin alteration may be necessary to rule out melanoma.


Depletion-controlled starvation of a diffusing forager.
We study the starvation of a lattice random walker in which each site initially contains one food unit and the walker can travel S steps without food before starving. When the walker encounters food, it is completely eaten, and the walker can again travel S steps without food before starving. When the walker hits an empty site, the time until the walker starves decreases by 1. In spatial dimension d=1, the average lifetime of the walker ⟨τ⟩∝S, while for d>2, ⟨τ⟩≃exp(S^{ω}), with ω→1 as d→∞; the latter behavior suggests that the upper critical dimension is infinite. In the marginal case of d=2, ⟨τ⟩∝S^{z}, with z≈2. Long-lived walks explore a highly ramified region so they always remain close to sources of food and the distribution of distinct sites visited does not obey single-parameter scaling.


The therapeutic potential of host-defense antimicrobial peptides.
Species across the evolutionary scale from insects to mammals deploy peptides to counter microbial invaders. Although, there are considerable differences in the primary and secondary structures of these peptides, they exert their antimicrobial activity by common mechanisms ie. by membrane permeabilization. Extensive structure-function studies have been carried out on this class of antimicrobial peptides and 'designer' peptides have been generated which possess specific antimicrobial activity. In this article the physico-chemical properties and biological activities of host-defense peptides and 'designer' peptides would be reviewed with a view to explore whether they could be used as effective therapeutic agents.


Surgical evolution in spinal dural arteriovenous fistula treatment-a 7 years monocentric experience.
Accounting for 70% of all spinal vascular malformations, spinal dural arteriovenous fistulas (SDAVF) are the most common type of malformation. Interruption of the fistulous arterialized vein point is the goal of surgical treatment. The aim of the study was to compare open surgery (laminectomy) versus minimal invasive surgery (MIS) in SDAVF treatment. Between March 2013 and March 2020, we retrospectively collected 21 consecutive adult patients with SDAVF. Since March 2017, MIS has been routinely used for surgical treatment. Pre- and post-operative clinical evaluations used Aminoff-Logue score (ALS). Complication rate was noted. Post-operative occlusion of the malformation was confirmed by digital subtraction angiography (DSA) in all patients. MIS was compared to open surgery in terms of efficacy and complications with statistical evaluation. Standard laminectomy was performed in 12 patients and MIS technique in 9 patients. No difference was noted on pre-operative parameters. ALS and MRI signs of myelopathy were improved in all cases except for 1 patient in each group. All SDAVFs were excluded based on post-operative DSA. Significant differences were noted between the 2 groups in terms of perioperative blood loss (p<0.001), post-operative pain visual analog scale values (p<0.001), and first time out of bed (p<0.001). Wrong level surgery occurred in one patient in each group; patients were re-operated using the same technique. No infection or cerebrospinal fluid (CSF) leak was noted. In our experience, MIS is a safe alternative to open laminectomy for SDAVF treatment. MIS contributes to patient comfort and minimizes blood loss without increasing complication rate.


Mono-, di- and trinuclear 2,3,5,6-tetrakis(2-pyridyl)pyrazine (tppz)-containing copper(II) complexes: syntheses, crystal structures and magnetic properties.
Three new copper(ii) complexes of formula [Cu(tppz)(NCO)(2)].0.4H(2)O (1), [Cu(2)(tppz)Br(4)](2) and [Cu(3)(tppz)(C(5)O(5))(3)(H(2)O)(3)].7H(2)O (3)[tppz = 2,3,5,6-tetrakis(2-pyridyl)pyrazine; C(5)O(5)(2-) = croconate, dianion of 4,5-dihydroxycyclopent-4-ene-1,2,3-trione] have been synthesised and structurally characterized by X-ray diffraction methods. The structure of complex is made up of neutral [Cu(tppz)(NCO)(2)] mononuclear units and uncoordinated water molecules. The mononuclear units are grouped by pairs to give a rather short copper-copper distance of 3.9244(4) angstroms. The structure of complex 1 consists of neutral tppz-bridged [Cu(2)(tppz)Br(4)] dinuclear units, the copper-copper separation across tppz being 6.6198(1) angstroms. The dinuclear units are further connected through weak, double out-of-plane Cu-Br...Cu bridges [Br(1)...Cu(1a) 4.0028(17) angstroms] creating tetranuclear entities, the copper-copper separation through this interaction being 4.3299(21) angstroms. The structure of complex 3 is built of neutral [Cu(3)(tppz)(C(5)O(5))(3)(H(2)O)(3)] trinuclear units and uncoordinated water molecules. Tppz and one of the croconate groups act as bridging ligands, the former exhibiting the bis-terdentate coordination mode and the latter adopting an unusual asymmetrical bis-bidentate bridging mode through three adjacent oxygen atoms. The other two croconate groups exhibit the bidentate coordination mode. The intramolecular copper-copper separations are 6.5417(9)(across tppz) and 4.3234(9) angstroms (through bis-bidentate croconato). The magnetic properties of 2 and 3 have been investigated in the temperature range 1.9-300 K. The magnetic behaviour of complex 2 is that of an antiferromagnetically coupled copper(II) dimer (J = -40.9 cm(-1), the Hamiltonian being H = -JS(A).S(B)). In the case of compound , the chi(M) T vs. T plot is typical of an overall antiferromagnetic coupling with a low-lying spin doublet being fully populated at T < 10 K. The values of the intramolecular antiferromagnetic interactions in 3 are -19.9 (across tppz) and -32.9 cm(-1)(through bridging croconato). Density functional type calculations were performed on model dinuclear fragments of 3 in order to analyze the efficiency of the exchange pathways involved and also to substantiate the coupling parameters.


Identification of genes involved in Ca2+ ionophore A23187-mediated apoptosis and demonstration of a high susceptibility for transcriptional repression of cell cycle genes in B lymphoblasts from a patient with Scott syndrome.
In contrast to other agents able to induce apoptosis of cultured cells, Ca2+ ionophore A23187 was shown to elicit direct activation of intracellular signal(s). The phenotype of the cells derived from patients having the hemorrhagic disease Scott syndrome, is associated with an abnormally high proportion of apoptotic cells, both in basal culture medium and upon addition of low ionophore concentrations in long-term cultures. These features are presumably related to the mutation also responsible for the defective procoagulant plasma membrane remodeling. We analyzed the specific transcriptional re-programming induced by A23187 to get insights into the effect of this agent on gene expression and a defective gene regulation in Scott cells. The changes in gene expression upon 48 hours treatment with 200 nM A23187 were measured in Scott B lymphoblasts compared to B lymphoblasts derived from the patient's daughter or unrelated individuals using Affymetrix microarrays. In a similar manner in all of the B cell lines, results showed up-regulation of 55 genes, out of 12,000 represented sequences, involved in various pathways of the cell metabolism. In contrast, a group of 54 down-regulated genes, coding for histones and proteins involved in the cell cycle progression, was more significantly repressed in Scott B lymphoblasts than in the other cell lines. These data correlated with the alterations of the cell cycle phases in treated cells and suggested that the potent effect of A23187 in Scott B lymphoblasts may be the consequence of the underlying molecular defect. The data illustrate that the ionophore A23187 exerts its pro-apoptotic effect by promoting a complex pattern of genetic changes. These results also suggest that a subset of genes participating in various steps of the cell cycle progress can be transcriptionally regulated in a coordinated fashion. Furthermore, this research brings a new insight into the defect in cultured Scott B lymphoblasts, leading to hypothesize that a mutated gene plays a role not only in membrane remodeling but also in signal transduction pathway(s) leading to altered transcriptional regulation of cell cycle genes.


Insulin in w/o/w multiple emulsions: preparation characterization and determination of stability towards proteases in vitro.
In this work two w/o/w multiple emulsions composed of soybean oil or medium-chain triglycerides and containing insulin were studied. These emulsions were prepared by means of two-step emulsification procedure. The w/o/w emulsions obtained were stable for at least 6 months of storage at 4-6 degrees C. The yield of encapsulation of insulin was > 95%. The main release mechanism is a swelling-breakdown phenomenon. In vitro, the two w/o/w multiple emulsions were able to protect insulin against enzymatic degradation. These results indicate that multiple emulsions have potential as a carrier of insulin for oral administration.


Esophageal Neuroendocrine Carcinoma With Focal Squamous Cell Carcinoma Component.
This study was about a 70-year-old man with progressive dysphagia. Esophagogastroduodenoscopy showed a 1.2-cm circumferential ulcerative mass at the level of 23 cm from the upper incisors in the upper esophagus. The mass was first diagnosed as a poorly differentiated squamous cell carcinoma. McKeown esophagogastrectomy was performed with intention to treat the lesion. The pathological examination showed an esophageal neuroendocrine carcinoma that was mixed with squamous cell carcinoma component. In this report, we present a unique case of the extremely rare esophageal neoplasm, an esophageal neuroendocrine carcinoma with squamous cell carcinoma component.


Targeting Gut Microbiota With Natural Polysaccharides: Effective Interventions Against High-Fat Diet-Induced Metabolic Diseases.
Unhealthy diet, in particular high-fat diet (HFD) intake, can cause the development of several metabolic disorders, including obesity, hyperlipidemia, type 2 diabetes mellitus (T2DM), non-alcoholic fatty liver disease (NAFLD), and metabolic syndrome (MetS). These popular metabolic diseases reduce the quality of life, and induce premature death worldwide. Evidence is accumulating that the gut microbiota is inextricably associated with HFD-induced metabolic disorders, and dietary intervention of gut microbiota is an effective therapeutic strategy for these metabolic dysfunctions. Polysaccharides are polymeric carbohydrate macromolecules and sources of fermentable dietary fiber that exhibit biological activities in the prevention and treatment of HFD-induced metabolic diseases. Of note, natural polysaccharides are among the most potent modulators of the gut microbiota composition. However, the prebiotics-like effects of polysaccharides in treating HFD-induced metabolic diseases remain elusive. In this review, we introduce the critical role of gut microbiota human health and HFD-induced metabolic disorders. Importantly, we review current knowledge about the role of natural polysaccharides in improving HFD-induced metabolic diseases by regulating gut microbiota.


Development and evaluation of brochures for fall prevention education created to empower nursing home residents and family members.
In this study, we describe the development of evidence- and theory-based fall prevention educational material and its evaluation from the users' perspectives. To reduce risk factors for falling in nursing homes, nursing staff must enact multifactorial fall prevention intervention programmes. A core component of these programmes is to educate residents and their family members, both verbally and in a written form. However, users can only benefit from educational material if it is based on current scientific evidence, easy to understand and process and customised. We followed a structured procedure during the development process, while considering various aspect of quality. To assess the understandability and usefulness of the resulting educational materials, we conducted a qualitative content analysis study. The educational materials development process incorporated several iterative steps including a systematic literature search and the application of frameworks for designing and writing the materials. To evaluate the material, we performed six focus group discussions separately with residents, family members and nursing staff from two nursing homes (total of 32 participants). Residents' brochures included clear information on avoiding external risks as well as coping strategies after a fall event. Family members' brochures were more comprehensive, including both concrete tips and outlining the advantages and disadvantages of interventions. Residents and family members had no difficulties understanding the material and tried to apply the content to their individual situations. Nursing staff commented on some ambiguities and incongruities relating to current nursing care practice. By involving users in the development of evidence-based educational materials, nursing staff can achieve a high acceptance rate for the materials and motivate users to address the topic. The involvement of users is essential for developing educational material that meets users' needs. Educational material should be used as part of an overall strategy to educate residents and family members in nursing homes.


Identifying a risk profile for thyroid cancer.
The large use of simple and effective diagnostic tools has significantly contributed to the increase in diagnosis of thyroid cancer over the past years. However, there is compelling evidence that most micropapillary carcinomas have an indolent behavior and may never evolve into clinical cancers. Therefore, there is an urgent need for new tools able to predict which thyroid cancers will remain silent, and which thyroid cancers will present an aggressive behavior. There are a number of well-established clinical predictors of malignancy and recent studies have suggested that some of the patients laboratory data and image methods may be useful. Molecular markers have also been increasingly tested and some of them appear to be very promising, such as BRAF, a few GST genes and p53 polymorphisms. In addition, modern tools, such as immunocytochemical markers, and the measure of the fractal nature of chromatin organization may increase the specificity of the pathological diagnosis of malignancy and help ascertain the prognosis. Guidelines designed to select nodules for further evaluation, as well as new methods aimed at distinguishing carcinomas of higher aggressiveness among the usually indolent thyroid tumors are an utmost necessity.


Identification of Angiotensin I-Converting Enzyme Inhibitory Peptides Derived from Enzymatic Hydrolysates of Razor Clam Sinonovacula constricta.
Angiotensin I-converting enzyme (ACE) inhibitory activity of razor clam hydrolysates produced using five proteases, namely, pepsin, trypsin, alcalase, flavourzyme and proteases from Actinomucor elegans T3 was investigated. Flavourzyme hydrolysate showed the highest level of degree of hydrolysis (DH) (45.87%) followed by A. elegans T3 proteases hydrolysate (37.84%) and alcalase (30.55%). The A. elegans T3 proteases was observed to be more effective in generating small peptides with ACE-inhibitory activity. The 3 kDa membrane permeate of A. elegans T3 proteases hydrolysate showed the highest ACE-inhibitory activity with an IC50 of 0.79 mg/mL. After chromatographic separation by Sephadex G-15 gel filtration and reverse phase-high performance liquid chromatography, the potent fraction was subjected to MALDI/TOF-TOF MS/MS for identification. A novel ACE-inhibitory peptide (VQY) was identified exhibiting an IC50 of 9.8 μM. The inhibitory kinetics investigation by Lineweaver-Burk plots demonstrated that the peptide acts as a competitive ACE inhibitor. The razor clam hydrolysate obtained by A. elegans T3 proteases could serve as a source of functional peptides with ACE-inhibitory activity for physiological benefits.


Catalytic stereoselective glycosidation with glycosyl diphenyl phosphates: rapid construction of 1,2-cis-alpha-glycosidic linkages.
A commercially available 0.1 M solution of HClO(4) in dioxane has been shown to catalyze the glycosidation of glycosyl diphenyl phosphates. The per-O-benzyl-protected glucosyl and galactosyl donors and the 3,4,6-tri-O-acetyl-2-azido-2-deoxygalactosyl donor each react with a range of acceptor alcohols in the presence of 0.05-0.2 equiv of HClO(4) in dioxane/Et(2)O (1:1) to afford glycosides in good yields with good to excellent alpha selectivities. The synthetic utility of this glycosidation method was demonstrated by a stereoselective synthesis of the alpha-galactosylceramide KRN7000, an activator of natural killer (NK) T cells through CD1d molecules.


Dissimilar effects of phorbol ester and diacylglycerol derivative on protein kinase activity in the monoblastoid U937 cell.
Mechanism, in addition to protein kinase C activation may mediate 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulated differentiation of leukemic cells. We compared the effect of pretreating intact monoblastoid U937 cells with TPA or the diacylglycerol derivative, 1-oleoyl-2-acetylglycerol (OAG), by studying the protein kinase C dependent and independent histone phosphotransferase activity, the phosphorylation of endogenous substrates, and the ability to stimulate differentiation. In cellular fractions derived from cells treated with TPA or OAG, cytosolic protein kinase C activity decreased. In the detergent extracted particulate fraction, TPA produced a time and dose dependent decrease in protein kinase C activity. In contrast, OAG increased particulate protein kinase C activity. In addition, the particulate fraction derived from cells treated with TPA exhibited increased phosphatidyl serine and diolein independent histone phosphotransferase activity as well as an increase in the phosphorylation of two endogenous substrates with molecular weights of 120,000 and 80,000. OAG did not mimic these effects. When exposed to 32P-labeled intact cells, OAG and TPA stimulated phosphorylation of three substrates. Thus, the inability of OAG to mimic the effects of TPA was not due to lack of protein kinase C activation. TPA, but not OAG, stimulated differentiation of the U937 cell to a monocyte-like cell. These data demonstrate that TPA and OAG have dissimilar effects on protein kinase activity and differentiation in the U937 monoblastoid cell.


Calcium Channels in Postnatal Development of Rat Pancreatic Beta Cells and Their Role in Insulin Secretion.
Pancreatic beta cells during the first month of development acquire functional maturity, allowing them to respond to variations in extracellular glucose concentration by secreting insulin. Changes in ionic channel activity are important for this maturation. Within the voltage-gated calcium channels (VGCC), the most studied channels are high-voltage-activated (HVA), principally L-type; while low-voltage-activated (LVA) channels have been poorly studied in native beta cells. We analyzed the changes in the expression and activity of VGCC during the postnatal development in rat beta cells. We observed that the percentage of detection of T-type current increased with the stage of development. T-type calcium current density in adult cells was higher than in neonatal and P20 beta cells. Mean HVA current density also increased with age. Calcium current behavior in P20 beta cells was heterogeneous; almost half of the cells had HVA current densities higher than the adult cells, and this was independent of the presence of T-type current. We detected the presence of α1G, α1H, and α1I subunits of LVA channels at all ages. The Cav 3.1 subunit (α1G) was the most expressed. T-type channel blockers mibefradil and TTA-A2 significantly inhibited insulin secretion at 5.6 mM glucose, which suggests a physiological role for T-type channels at basal glucose conditions. Both, nifedipine and TTA-A2, drastically decreased the beta-cell subpopulation that secretes more insulin, in both basal and stimulating glucose conditions. We conclude that changes in expression and activity of VGCC during the development play an important role in physiological maturation of beta cells.


A Drosophila functional evaluation of candidates from human genome-wide association studies of type 2 diabetes and related metabolic traits identifies tissue-specific roles for dHHEX.
Genome-wide association studies (GWAS) identify regions of the genome that are associated with particular traits, but do not typically identify specific causative genetic elements. For example, while a large number of single nucleotide polymorphisms associated with type 2 diabetes (T2D) and related traits have been identified by human GWAS, only a few genes have functional evidence to support or to rule out a role in cellular metabolism or dietary interactions. Here, we use a recently developed Drosophila model in which high-sucrose feeding induces phenotypes similar to T2D to assess orthologs of human GWAS-identified candidate genes for risk of T2D and related traits. Disrupting orthologs of certain T2D candidate genes (HHEX, THADA, PPARG, KCNJ11) led to sucrose-dependent toxicity. Tissue-specific knockdown of the HHEX ortholog dHHEX (CG7056) directed metabolic defects and enhanced lethality; for example, fat-body-specific loss of dHHEX led to increased hemolymph glucose and reduced insulin sensitivity. Candidate genes identified in human genetic studies of metabolic traits can be prioritized and functionally characterized using a simple Drosophila approach. To our knowledge, this is the first large-scale effort to study the functional interaction between GWAS-identified candidate genes and an environmental risk factor such as diet in a model organism system.


Origin of the genetic code: a testable hypothesis based on tRNA structure, sequence, and kinetic proofreading.
We hypothesize that the origin of the genetic code is associated with the structure of the tRNA that existed in primal cells. The sequences of modern tRNA contain correlations which can be understood as "fossil" evidence of the secondary structure of primal tRNA. Kinetic proofreading through diffusion can amplify a low level of intrinsic selectivity of tRNA for its amino acid. Experimental tests of the theory are suggested.


OMICs approaches and technologies for understanding low-high feed efficiency traits in chicken: implication to breeding.
In poultry production, there has been a trend of continuous increase in cost of feed ingredients which represents the major proportion of the production costs. Feed costs can be reduced by improving feed efficiency traits which increase the possibility of using various indigestible feed sources and decrease the environmental impact of the enhanced poultry production. Therefore, feed efficiency has been used as one of the most important economic traits of selection in the breeding program of chickens. Recently, many OMICs experimental studies have been designed to characterize biological differences between the high and low feed efficiency chicken phenotypes. Biological complexity cannot be fully captured by main individual OMICs such as genomics, transcriptomics, proteomics and metabolomics. Therefore, researchers have combined multiple assays from the same set of samples to create multi-OMICs datasets. OMICs findings are crucial in improving existing approaches to poultry breeding. The current review aimed to highlight the components of feed efficiency and general OMICs approaches and technologies. Besides, individual and multi-OMICs based understanding of chicken feed efficiency traits and the application of the acquired knowledge in the chicken breeding program were addressed.


Artificial Intelligence Algorithm-Based Magnetic Resonance Imaging to Evaluate the Effect of Radiation Synovectomy for Hemophilic Arthropathy.
This study aimed to discuss magnetic resonance imaging (MRI) based on artificial intelligence (AI) algorithm to evaluate the effect of radiation synovectomy for hemophilic arthropathy (HA). MRI based on the Canny algorithm was applied and compared with conventional MRI to evaluate its application effects according to the PSNR and SSIM. Sixty patients diagnosed with HA were selected as the research subjects. According to the detection method, the patients were divided into group A (pathological detection after radiation synovectomy), group B (conventional MRI detection), and group C (MRI detection based on the Canny algorithm). The application value of MRI based on the Canny algorithm was judged by comparing the differences between the two detection methods and pathological results. The results displayed that the reconstruction effect of the Canny algorithm was remarkably better than that of the traditional algorithm regarding the peak signal-to-noise ratio (PSNR) and structural similarity (SSIM), which showed a clearer synovial contour. The results of the IPSG score of joint effusion and hemorrhage showed that there was a difference in the detection rate of joints between conventional MRI and pathological results on the score of 1 and 2 (P < 0.05); and there was no significant difference between the MRI and pathological results based on the Canny algorithm (P > 0.05). The results of the IPSG score of synovial hyperplasia showed that the detection rate of conventional MRI was different from pathological results on the score of 1 and 2 (P < 0.05); and there was no significant difference between the MRI and pathological results based on the Canny algorithm (P > 0.05). The results of the IPSG score of hemosiderin deposition showed that the detection rate of conventional MRI was different from the pathological results on the score of 1 and 2 (P < 0.05); and there was no significant difference between the MRI and pathological results based on the Canny algorithm (P > 0.05). The synovial volume of patients after surgery was reduced compared with that before surgery. One-factor variance was used to analyze the clinical hemorrhage frequency before and after surgery, and the results showed that the differences were statistically significant (P < 0.05). Therefore, MRI on account of AI algorithm made it easier to detect synovial contour, which was helpful to evaluate the efficacy of polygenic risk scores (PRS) surgery in HA patients. MRI based on the Canny algorithm had less differences between the score of hemophilic arthropathy and pathological results, which could replace conventional MRI examination and have clinical application value.


[Biochemical indices of connective tissue metabolism in patients with chronic hepatitis B and C].
37 patients with chronic hepatitis B and C were examined. Patients were divided into 3 groups depending on the degree of connective tissue dysplasia. We investigated: free and protein-bounded hydroxyproline, collagenase activity, total alkaline phosphatase and its bone fraction, creatinine, calcium and phosphorus content in the blood serum and urine. It has been found the dependence of collagen synthesis from the state of connective tissue. The higher is the degree of dysplasia, the more intensive is the process of collagen synthesis (P < 0.05). The index of corellation between protein-bounded and free fraction can be used as a biochemical marker for determination the stage of pathological process in the liver and for monitoring the effectiveness of therapy.


A comprehensive evaluation of application of light-cured colored compomer for deciduous teeth.
Twinky Star colored compomer produced by VOCO GmbH is a new type of filling material designed with seven different colors. Previous studies have shown that using the colored filling material can induce children's curiosity and increase the acceptance for dental treatment. However, few studies have been conducted to evaluate it. Hence, the present study was designed to assess the mechanical and adhesive properties of this new compomer, examine its biological safety, and evaluate the effects of its clinical application on children's adherence to dental treatment and the subsequent therapeutic outcomes. We found that Twinky Star compomer has a high compressive strength, good biaxial flexural strength, low wear rate, and good adhesive properties, and had little in vitro cytotoxic effects and did not cause apparent hemolysis in vivo. Finally, application of the compomer helped to improve children's acceptance of treatment without affecting the subsequent therapeutic outcomes in the follow-up examinations. To summarize, the findings suggest that Twinky Star compomer meets the performance requirements for dental fillings, and can be recommended for future clinical application.


Cluster Analysis of Characteristics of Persons Who Died by Suicide in the Montreal Metro Transit.
Suicides occur in metro systems worldwide and patterns vary in different urban transit networks. This study presents an in-depth analysis of 117 suicides in the Montreal Metro from 2000 to 2008 based on data obtained from coroners' investigations. Cluster analyses were performed to identify characteristics of groups of people who kill themselves in the Montreal Metro. We also compared changes in characteristics with data from 1986 to 1995. We identified five clusters of suicidal persons that describe patterns of characteristics of individuals who died by suicide in the metro that may be useful for prevention. Comparisons of suicides during 2000-2008 with data from a previous study of coroners' investigations of Montreal Metro suicides during 1986-1995 indicate changes in age, isolation, types of problems, and geographic patterns. Characteristics of metro suicides may be specific to localities and can change over time. Their understanding may facilitate the development of prevention strategies tailored to these different profiles.


Salvage of total esophageal reconstruction with microvascular free left colon flap.
Esophageal reconstruction has been achieved with a variety of innovative surgical techniques, including colonic interposition, gastric pull-up, pedicled, and free jejunal grafts. The authors describe an unusual case of complete thoracic esophageal reconstruction, necessitating an entirely free microvascular colon interposition as a salvage procedure to reconstitute the alimentary tract from the neck to the abdomen. To their knowledge, this specific method of reconstruction of the esophagus has not been previously reported.


Structure-based identification and characterisation of structurally novel human P2X7 receptor antagonists.
The P2X7 receptor (P2X7R) plays an important role in diverse conditions associated with tissue damage and inflammation, meaning that the human P2X7R (hP2X7R) is an attractive therapeutic target. The crystal structures of the zebrafish P2X4R in the closed and ATP-bound open states provide an unprecedented opportunity for structure-guided identification of new ligands. The present study performed virtual screening of ∼100,000 structurally diverse compounds against the ATP-binding pocket in the hP2X7R. This identified three compounds (C23, C40 and C60) out of 73 top-ranked compounds by testing against hP2X7R-mediated Ca(2+) responses. These compounds were further characterised using Ca(2+) imaging, patch-clamp current recording, YO-PRO-1 uptake and propidium iodide cell death assays. All three compounds inhibited BzATP-induced Ca(2+) responses concentration-dependently with IC50s of 5.1±0.3μM, 4.8±0.8μM and 3.2±0.2μM, respectively. C23 and C40 inhibited BzATP-induced currents in a reversible and concentration-dependent manner, with IC50s of 0.35±0.3μM and 1.2±0.1μM, respectively, but surprisingly C60 did not affect BzATP-induced currents up to 100μM. They suppressed BzATP-induced YO-PRO-1 uptake with IC50s of 1.8±0.9μM, 1.0±0.1μM and 0.8±0.2μM, respectively. Furthermore, these three compounds strongly protected against ATP-induced cell death. Among them, C40 and C60 exhibited strong specificity towards the hP2X7R over the hP2X4R and rP2X3R. In conclusion, our study reports the identification of three novel hP2X7R antagonists with micromolar potency for the first time using a structure-based approach, including the first P2X7R antagonist with preferential inhibition of large pore formation.


Tooth formation and the 28,000-dalton vitamin D-dependent calcium-binding protein: an immunocytochemical study.
Antiserum to the 28-kilodalton vitamin D-dependent calcium-binding protein (CaBP) was used to localize CaBP in histologic sections of the continuously erupting incisor in mandibles obtained from normal rats. With the peroxidase--anti-peroxidase technique, no CaBP was detected in undifferentiated ameloblasts or in those which had become columnar and were facing pulp. Calcium-binding protein was first noted in the cytoplasm of random ameloblasts facing dentin in the presecretion zone. As the ameloblasts became more mature in the zone of enamel secretion, CaBP was uniformly present in their cytoplasm. Ameloblasts with Tome's processes clearly contained CaBP in these processes as well as in the cell-body cytoplasm. Near the later developmental stages of the zone of enamel secretion, some of the adjacent underlying cells of the stratum intermedium also contained CaBP in their cytoplasm. In some stratum intermedium cells and papillary cells, CaBP extended into the zone of enamel maturation, but not to the end of that zone. Cytoplasmic CaBP continued to be present in ameloblasts as they progressed through the zone of enamel maturation to the final, shortened cells at the gingival margin of the erupting incisor. No CaBP was detected in odontoblasts, pulpal cells, the stellate reticulum, or the outer dental epithelium.


Brief Report: Biological Sound Processing in Children with Autistic Spectrum Disorder.
There is debate whether social impairments in autism spectrum disorder (ASD) are truly domain-specific, or if they reflect generalized deficits in lower-level cognitive processes. To solve this issue, we used auditory-evoked EEG responses to assess novelty detection (MMN component) and involuntary attentional orientation (P3 component) induced by socially-relevant, human-produced, biological sounds and acoustically-matched control stimuli in children with ASD and controls. Results show that early sensory and novelty processing of biological stimuli are preserved in ASD, but that automatic attentional orientation for biological sounds is markedly altered. These results support the notion that at least some cognitive processes of ASD are specifically altered when it comes to processing social stimuli.


A Framework for Utilizing High-Resolution Mass Spectrometry and Nontargeted Analysis in Rapid Response and Emergency Situations.
Unknown chemical releases constitute a large portion of the rapid response situations to which the US Environmental Protection Agency is called on to respond. Workflows used to address unknown chemical releases currently involve screening for a large array of known compounds using many different targeted methods. When matches are not found, expert analytical chemistry knowledge is used to propose possible candidates from the available data, which generally includes low-resolution mass spectra and situational clues such as the location of the release, nearby industrial operations, and other field-reported facts. The past decade has witnessed dramatic improvements in capabilities for identifying unknown compounds using high-resolution mass spectrometry (HRMS) and nontargeted analysis (NTA) approaches. Complementary developments in cheminformatics tools have further enabled an increase in NTA throughput and identification confidence. Together with the expanding availability of HRMS instrumentation in monitoring laboratories, these advancements make NTA highly relevant to rapid response scenarios. In this article, we introduce the concept of NTA as it relates to rapid response needs and describe how it can be applied to address unknown chemical releases. We advocate for the consideration of HRMS-based NTA approaches to support future rapid response scenarios. Environ Toxicol Chem 2022;41:1117-1130. Published 2021. This article is a U.S. Government work and is in the public domain in the USA.


Hospital Prices Increase in California, Especially Among Hospitals in the Largest Multi-hospital Systems.
A surge in hospital consolidation is fueling formation of ever larger multi-hospital systems throughout the United States. This article examines hospital prices in California over time with a focus on hospitals in the largest multi-hospital systems. Our data show that hospital prices in California grew substantially (+76% per hospital admission) across all hospitals and all services between 2004 and 2013 and that prices at hospitals that are members of the largest, multi-hospital systems grew substantially more (113%) than prices paid to all other California hospitals (70%). Prices were similar in both groups at the start of the period (approximately $9200 per admission). By the end of the period, prices at hospitals in the largest systems exceeded prices at other California hospitals by almost $4000 per patient admission. Our study findings are potentially useful to policy makers across the country for several reasons. Our data measure actual prices for a large sample of hospitals over a long period of time in California. California experienced its wave of consolidation much earlier than the rest of the country and as such our findings may provide some insights into what may happen across the United States from hospital consolidation including growth of large, multi-hospital systems now forming in the rest of the rest of the country.


Three-dimensional polarization effects in optical tunneling.
We consider the three-dimensional (3D) polarimetric properties of an evanescent optical field excited in the gap of a double-prism system by a random plane wave. The analysis covers the case of frustrated total internal reflection (FTIR), i.e., optical tunneling, and relies on the characteristic decomposition of the 3×3 polarization matrix. We find in particular that, for any incident partially polarized plane wave, the evanescent field inside the gap is necessarily in a nonregular, genuine 3D polarization state. We also show that the 3D polarimetric properties of the field at the second boundary are sensitive to the changes of the gap width and that the relevant effects occur for the smaller widths when the angle of incidence of the plane wave becomes larger. The results of this work uncover new aspects of the polarimetric structure of genuine 3D evanescent fields and may find applications in near-field optics and surface nanophotonics.


Lattice kinetic simulations in three-dimensional magnetohydrodynamics.
A lattice kinetic algorithm to simulate three-dimensional (3D) incompressible magnetohydrodynamics is presented. The fluid is monitored by a distribution function, which obeys a scalar kinetic equation, subject to an external force due to the imposed magnetic field. Following the work of J. Comput. Phys. 179, 95 (2002)], the magnetic field is represented by a different three-component vector distribution function, which obeys a corresponding vector kinetic equation. Discretization of the 3D phase space is based on a 19-bit scheme for the hydrodynamic part and on a 7-bit scheme for the magnetic part. Numerical results for magnetohydrodynamic (MHD) flow in a rectangular duct with insulating and conducting walls provide excellent agreement with corresponding analytical solutions. The scheme maintains in all cases tested the MHD constraint inverted Delta.B=0 within machine round-off error.


Assessing cross-cultural equivalence through confirmatory factor analysis.
This article presents a study in which Picot's caregiver rewards scale (PCRS), originally developed in English, was cross-culturally validated with 137 Chinese adult children family caregivers in the United States using confirmatory factor analysis. A one-factor structure of the 21-item revised Chinese PCRS was supported as indicated by goodness-of-fit index = .94, adjusted goodness-of-fit index = .93, standardized root mean square residual = .09, and chi-square to df ratio = 2.7. Chi-square for this model was (chi(2) [189, n = 137] = 514, p < .05). The standardized alpha was .90. All factor loadings were significant (p < .00) and above .30. Construct validity was supported by significant associations with caregivers' filial beliefs (r = .32, p < .01) and caregivers' filial affection toward parents (r = .23, p < .05). Results suggest that the Chinese PCRS is a summative measure of perceived caregiver rewards with potential for evaluating interventions to reduce perceived burden among adult children family caregivers.


Weight loss maintenance for 2 years after a 6-month randomised controlled trial comparing education-only and group-based support in Japanese adults.
Our previous study, a 6-month randomised controlled trial, demonstrated that a group-based support promoted weight loss as compared to an education-only intervention. The purpose of this study was to examine weight loss maintenance for 2 years. Originally, 188 overweight Japanese adults, aged 40-65 years, were randomly assigned to 3 groups: control, education-only or group-based support. After the 6-month intervention, 125 participants in the education-only and the group-based support groups were followed up for 2 years. The primary outcome was the amount of weight lost. The participants were retrospectively grouped into quartiles of percent weight loss for secondary analyses. At the end of follow-up, the amount of weight lost in the education-only and the group-based support groups was the same (3.3 kg). Secondary analyses using data of those who completed the study (n = 100) revealed that the participants in the highest quartile of percent weight loss significantly increased their step counts and moderate-to-vigorous physical activity compared with the lowest quartile. No significant differences were observed in the energy intake among the four groups. The effects of group-based support disappear within 2 years. Increasing physical activity may be a crucial factor for successful maintenance of weight loss.


Chemotherapy of advanced ovarian cancer.
Majority of ovarian cancer patients have advanced disease (stage III or IV) at diagnosis and the prognosis of these patients is poor in spite of aggressive surgery. Therefore chemotherapy has gained a fundamental role in the therapeutic approach of ovarian cancer. Platinum compounds in combination with alkylating agents and taxoids have the higher antitumor activity in ovarian cancer, while the role of anthracyclines remains controversial. Our 10-year experience with cisplatinum-based polychemotherapy in 196 advanced ovarian cancer patients previously untreated with chemotherapy is reported. 74 patients were treated with the combination cis-platinum and anthracyclines; 53 patients received the combination cis-platinum plus epirubicin alternated to cyclophosphamide plus 5-fluorouracil; 48 patients were treated with cis-platinum plus cyclophosphamide plus epirubicin and 21 patients were treated with the same combination with intraperitoneal administration of cisplatin. Our data confirm literature results of 55% remission rate, with 29% showing complete remissions. The median survival was 79 weeks and the overall 10-year survival was 13%. Complete responders had a median survival of 263 weeks and a 30% survival at 10 years. The main prognostic factors in our retrospective analysis were the objective remission, the size of residual tumor, the performance status and the stage. With the combination carboplatin (300-400 mg/sm) and cyclophosphamide (600 mg/sm) we observed 80% objective responses (23% complete responses) in 53 advanced ovarian cancer patients. The median overall survival in this group was 140 weeks. We carried out a phase II, non-randomized study of taxol in 54 ovarian cancer patients pretreated with platinum-compounds. The overall tolerability was good and an objective remission was observed in 47% of cases (8% complete remissions). The median survival was 68 weeks. As a consequence of our previous experience, a phase I dose-finding study with the combination carboplatin and taxol was started in our Division in 1994. Up to now, 22 chemotherapy untreated patients entered the study and the 5th dose level (taxol 175 mg/sm and carboplatin 350 mg/sm) has been completed without reaching the maximum tolerated dose. Our preliminary data suggest that the combination taxol-carboplatin is very active as the first-line chemotherapy in advanced ovarian cancer (73% objective remissions in 15 evaluated patients).


Influence of age on saline hypertension in subtotal nephrectomized rats.
In uninephrectomised immature and adult male rats 34% renal tissue was removed from the remaining kidney and after 60-days exposure to saline treatment (0.17 mol/l NaCl solution as only drinking fluid) the mean arterial blood pressure, plasma urea concentration, plasma and extracellular fluid volumes were estimated. In comparison with water drinking uninephrectomised age-matched controls it has been found that: in both age groups, the loss of tissue from the remaining kidney was fully replaced by compensatory growth of the renal stump, plasma urea concentration remained unchanged in animals operated on when adult, but increased in animals operated on when immature, the interstitial fluid volume increased in both age groups--the plasma volume as well as blood pressure remained unchanged in animals treated when adult, but increased in animals treated when immature. It is concluded that under conditions of elevated salt intake the loss of renal mass in immature rats was compensated by growth of tissue with a lower excretory ability than in adult ones, this being responsible for the development of hypertension in the younger group.


Size-dependent spectroscopic properties of conjugated polymer nanoparticles.
This paper is focused on how the spectroscopic properties of conjugated polymers evolve in the size range between single polymer chains and the bulk material. The measurements used single-particle spectroscopy techniques and include both static and dynamic measurements. The main observation of this work is that the spectroscopic properties of MEH-PPV evolve rapidly as a function of nanoparticle size and achieve bulk-like properties for nanoparticles greater than 10 nm in size. Nanoparticles were assembled by a reprecipitation technique and characterized by fluorescence emission spectroscopy. The physical origin of the size-dependent spectroscopic properties is assigned to the distance dependence of four main processes: electronic energy transfer between blue and red sites, triplet-triplet annihilation, singlet exciton quenching by triplets, and singlet exciton quenching by hole polarons.


Apolipoprotein B and triacylglycerol secretion in human triacylglycerol hydrolase transgenic mice.
Apolipoprotein B (apoB)-containing lipoproteins play a critical role in whole body lipid homeostasis and the pathogenesis of atherosclerosis. The assembly of hepatic apoB-containing lipoproteins, VLDL, is governed by the availability of lipids, including triacylglycerol (TG). The majority of TG associated with VLDL is derived from the hepatic cytoplasmic lipid stores by a process involving lipolysis followed by reesterification. Microsomal triacylglycerol hydrolase (TGH) has been demonstrated to play a role in the lipolysis/reesterification process. To evaluate the potential regulatory role of TGH in hepatic VLDL assembly, we developed inducible transgenic mice expressing a human TGH minigene under the control of the mouse metallothionein promoter. Induction of human TGH by zinc resulted in liver-specific expression of the enzyme associated with 3- to 4-fold increases in lipolytic activity that could be attenuated with a TGH-specific inhibitor. Augmented TGH activity led to increased secretion of newly synthesized apoB and plasma TG levels. These results suggest that increased hepatic expression of TGH leads to a more proatherogenic plasma lipid and apoB profile.


Maritime health: a review with suggestions for research.
International maritime health has largely developed within the sphere of occupational health services and international health problems. We reviewed publications in the journal International Maritime Health from 2000 to 2010 to establish the coverage of the journal and the scope of research in maritime health. We identified six thematic categories: healthcare access, delivery and integration; telehealth; non-communicable diseases and physical health problems; communicable diseases; psychological functioning and health; and safety-related issues. We describe the research within these themes and report on their publication prominence. We also analyse the research in terms of its geographical focus, the population groups addressed and the research methodologies used. We suggest a broadening of maritime research to include randomised controlled trials, longitudinal studies and more qualitative research; more research addressing the context for non-European seafarers; and research on seafarers spouses and family supports and obligations. We also recommend more research on psychosocial and cultural issues and on telehealth, as well as the development of a stronger systems perspective for promoting maritime health.


Metabolism of chloroethanes by rat liver nuclear cytochrome P-450.
1,2-Dichloroethane, 1,1,1-trichloroethane and 1,1,2,2-tetrachloroethane appear to be metabolized by hepatic nuclear cytochrome P-450. All of these compounds are converted to chlorinated metabolites after incubation with hepatic nuclei and an NADPH-generating system plus EDTA, with the omission of any component eliminating metabolite production. In addition, CO, an inhibitor of cytochrome P-450, diminished the production of the chlorinated metabolites by hepatic nuclear preparations. The major metabolites of the chlorinated ethanes from hepatic microsomal cytochrome P-450, viz. chloroacetaldehyde from 1,2-dichloroethane, 2,2,2-trichloroethanol from 1,1,1-trichloroethane, and dichloroacetic acid from 1,1,2,2-tetrachloroethane, were also produced from the three chloroalkanes by hepatic nuclear cytochrome P-450. The levels of the metabolites produced were 65, 0.09 and 4.4 nmol/nmol cytochrome P-450/60 min. It is proposed that the pathways for the formation of these metabolites by hepatic nuclear cytochrome P-450 are as for their production by hepatic microsomal cytochrome P-450. Chloral hydrate was produced from 1,1,1-trichloroethane by hepatic nuclei plus NADPH, but not by hepatic microsomes. The presence of reactive species or transient enzyme bound intermediates in the pathways for the cytochrome P-450 dependent metabolism of the chloroethanes in hepatic nuclei is suggested by the observation that nuclear cytochrome P-450 is degraded in the presence of the chloroethanes in a NADPH dependent process which is inhibited by CO. It is proposed that, although the cytochrome P-450 dependent metabolism of the chloroethanes in microsomes can greatly exceed that in nuclei, the metabolism of 1,2-dichloroethane and 1,1,2,2-tetrachloroethane by nuclear cytochrome P-450 may in part mediate the mutagenicity and carcinogenicity of parent compounds.


[Isolation and identification of the infectious pancreatic necrosis virus in trout].
The first isolation and identification of the infectious pancreatic necrosis virus in trout in Czechoslovakia is described. The RTG-2, PG and CHSE cell lines were used for isolation and the identification was made by the methods of electron microscopy, cross virus-neutralizing test, and immunofluorescence. As demonstrated, all the isolates in the Czechoslovak territory are serologically identical and are closely related (or perhaps congruent) with the Sp reference strain which has the highest frequency in Europe. The described methodical procedure is also applicable to the diagnostics of other virus diseases of fish.


Prevalence, antimicrobial resistance and genetic diversity of Yersinia enterocolitica isolated from retail frozen foods in China.
In this study, our aim was to estimate the extent of Yersinia enterocolitica contamination in frozen foods in China and determine the bioserotype, virulotype, antimicrobial resistance, and enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) genotyping profiles of recovered Y. enterocolitica isolates. Out of 455 samples collected between July 2011 and May 2014, 56 (12.3%) tested positive for Y. enterocolitica. The 70 isolated strains were grouped into five clusters and one singleton based on their ERIC-PCR fingerprint, at a similarity coefficient of 70%. All strains were of biotype 1A, and 35.7% were of bioserotype 1A/O:8. Most strains lacked the virulence genes ail, virF, ystA, and ystC, but harbored ystB, fepD, ymoA, fes and sat. All strains were sensitive to ticarcillin but resistant to two or more antibiotics, and 48.6% of the strains were resistant to four to nine antibiotics. High resistance rates were observed for ampicillin, cephalothin, trimethoprim/sulfamethoxazole, amoxicillin/clavulanic acid, nalidixic acid and chloramphenicol (98.6%, 95.7%, 74.3%, 28.6%, 18.6% and 12.9%, respectively). This study provides a systematic surveillance of Y. enterocolitica prevalence in frozen foods in China and indicates its high antibiotic resistance, which could serve as useful information for the government to control Y. enterocolitica contamination in frozen foods and the use of antibiotics.


Evaluation of the performance of nucleic acid amplification tests (NAATs) in detection of chlamydia and gonorrhoea infection in vaginal specimens relative to patient infection status: a systematic review.
We evaluated the performance of nucleic acid amplification tests (NAATs) using vaginal specimens in comparison to specimens from the cervix or urine in their ability to detect chlamydia and gonorrhoea infection in women based on patient infection status (PIS). Systematic review. EMBASE and Ovid MEDLINE databases were searched through 3 October 2017. We included studies that tested samples from the vagina and ≥1 other site (cervix and/or urine) with ≥2 NAATs for chlamydia and ≥2 NAATs or 1 NAAT and culture for gonorrhoea for each site. Performance is defined as the sensitivity of a NAAT using a specimen site and PIS of the patient. We assessed risk of bias using modified QUADAS-2. Nine publications met the inclusion criteria (eight for chlamydia; six for gonorrhoea) and were narratively reviewed. Pooled summary estimates were not calculated due to the variable methodology and PIS definitions. Tests performed on vaginal specimens accomplished similar performance to cervical and urine specimens for chlamydia (range of performance estimates: vaginal 65%-100%, cervical 59%-97%, urine 57%-100%) and gonorrhoea (vaginal 64%-100%, cervical 85%-100%, urine 67%-94%). Vaginal specimens were estimated to have a performance >80% for chlamydia and gonorrhoea infections in all but one study. Performance of the NAATs for chlamydia and gonorrhoea detection using vaginal specimens was similar to that of cervical and urine specimens relative to PIS. As vaginal samples have a higher acceptability and lower cost, the study can support clinical testing guidelines by providing evidence that vaginal samples are a suitable alternative to traditionally used specimens.


Association of dietary glycemic index and glycemic load with endometrial cancer risk among Chinese women.
We evaluated the association of dietary glycemic index (GI) and glycemic load (GL) with the risk of endometrial cancer in a population-based, case-control study of 1199 endometrial cancer patients and 1212 age-frequency-matched controls in urban Shanghai, China, where diets are typically high in carbohydrates and have a high GL. Information on dietary habits, physical activity, and other relevant information was collected using a validated questionnaire, and anthropometric measurements were taken. Logistic regression was applied in the analysis. Dietary GI and GL were independently associated with risk for endometrial cancer but carbohydrate intake was unrelated to risk. Multivariable-adjusted odds ratios (ORs) for increasing quartiles of intake were 1.0, 1.3, 1.4, and 2.2 [95% confidence interval (CI): 1.2-4.0] for dietary GL (P(trend) = 0.02) and 1.0, 1.2, 1.4, and 1.4 (95% CI: 1.0-2.0) for dietary GI (P(trend) = 0.02). High intake of staples, especially rice, was positively associated with endometrial cancer. The association with GI was more evident among lean and normal weight women, although the test for interaction was not significant. This study suggests that intake of high GL or GI foods, but not carbohydrates per se, may increase risk for endometrial cancer.


[Post-tonsillectomy hemostasis: a prospective, randomized, double-blind study comparing argon beam coagulation and monopolar electrocoagulation].
Argon Beam Coagulation (ABC) enables effective hemostasis and limited penetration depth of the coagulation zone. This prospective, randomized, double-blind study compares ABC and monopolar electrocoagulation for post-tonsillectomy hemostasis. Fifty patients undergoing elective tonsillectomy were enrolled into the study. Final hemostasis was achieved using ABC in one fossa and electrocoagulation in the other fossa, with the sides chosen at random. Postoperative bleeding, postoperative pain and the healing of the tonsillar fossa were compared for both methods. No differences in any of the outcome measures were found between the two methods. ABC was felt to be faster and more comfortable than monopolar electrocoagulation. ABC is an effective and comfortable method for post-tonsillectomy hemostasis.


Evaluation of multi-echo ICA denoising for task based fMRI studies: Block designs, rapid event-related designs, and cardiac-gated fMRI.
Multi-echo fMRI, particularly the multi-echo independent component analysis (ME-ICA) algorithm, has previously proven useful for increasing the sensitivity and reducing false positives for functional MRI (fMRI) based resting state connectivity studies. Less is known about its efficacy for task-based fMRI, especially at the single subject level. This work, which focuses exclusively on individual subject results, compares ME-ICA to single-echo fMRI and a voxel-wise T2(⁎) weighted combination of multi-echo data for task-based fMRI under the following scenarios: cardiac-gated block designs, constant repetition time (TR) block designs, and constant TR rapid event-related designs. Performance is evaluated primarily in terms of sensitivity (i.e., activation extent, activation magnitude, percent detected trials and effect size estimates) using five different tasks expected to evoke neuronal activity in a distributed set of regions. The ME-ICA algorithm significantly outperformed all other evaluated processing alternatives in all scenarios. Largest improvements were observed for the cardiac-gated dataset, where ME-ICA was able to reliably detect and remove non-neural T1 signal fluctuations caused by non-constant repetition times. Although ME-ICA also outperformed the other options in terms of percent detection of individual trials for rapid event-related experiments, only 46% of all events were detected after ME-ICA; suggesting additional improvements in sensitivity are required to reliably detect individual short event occurrences. We conclude the manuscript with a detailed evaluation of ME-ICA outcomes and a discussion of how the ME-ICA algorithm could be further improved. Overall, our results suggest that ME-ICA constitutes a versatile, powerful approach for advanced denoising of task-based fMRI, not just resting-state data.


Performance Analysis for SWIPT Cooperative DF Communication Systems with Hybrid Receiver and Non-Linear Energy Harvesting Model.
In this paper, we study the outage and throughput performance for the simultaneous wireless information and power transfer (SWIPT) cooperative decode-and-forward (DF) communication systems. The hybrid receiver that uses both time switching (TS) and power splitting (PS) methods for energy harvesting (EH) and information decoding (ID), and the piece-wise linear EH model that captures the non-linear input-output characteristic of the EH circuit, are considered. We present exact analytical expressions of the outage probability (OP) and throughput, which are expressed as single definite integral on finite interval and can be easily evaluated, for the systems in Rayleigh fading channel. For further simplicity of calculation, we derive novel and closed-form approximate expressions of the OP and throughput. The impact of different system parameters on the system performance is investigated. Numerical results show the high accuracy of the proposed closed-form approximate expressions especially in the region of higher signal-to-noise ratio (SNR). It is also shown that the system performance is greatly overestimated when the ideal linear EH model is used instead of the practical non-linear EH model. A different result to the non-hybrid receiver with both linear EH model and non-linear EH model that there exists an optimal location to minimize the OP for the hybrid receiving relay node with non-linear EH model is also demonstrated.


Endovascular brachytherapy: overcoming "practical" obstacles.
Endovascular radiation therapy has proved to be safe and effective in preventing restenosis after coronary intervention both in animal studies and in pilot feasibility studies in humans. The rationale for such therapy is that radiation prevents neointimal proliferation and vessel constriction after vascular injury. Two forms of endovascular radiation therapy are available: catheter-based systems and radioactive stenting. However, several practical issues related to this technique still need to be addressed, such as questions about dosimetry, shielding, expense, handling and disposal of radioisotopes, certification for potential users, and-above all-safety and efficacy. In their pivotal studies, investigators and industry need to focus on prioritizing the clinical applications of this form of treatment. The clinical trials must attempt to determine the cost-effectiveness of this therapy as well as its risk versus benefit. For example, brachytherapy may prove to be of great benefit to patients with in-stent restenosis, and favorable results of studies examining this application should expedite the approval of endovascular radiation therapy for clinical use.


Child abuse: recognition and reporting by health professionals.
Nurses who are involved in child abuse and neglect have three major areas of concern: the child, the parent or caretaker, and their personal feelings. In terms of the child, the nurse needs to know the physical and behavioral indicators of abuse and neglect in order to make objective assessments. Indicators may be mild or severe, and usually occur in a pattern or in combination. The abused child may evoke sympathy in the nurse to an extent that may be detrimental to the resolution of a family problem. In terms of the parent, the nurse needs to determine the presence or absence of mental pathology and to recognize the good intent of the majority of abusive and neglectful parents. In dealing with parents, the nurse must always be honest, nonjudgmental, and supportive. Discussion of the report offers the best opportunity for providing support. Personal feelings are the greatest obstacle to the successful management of child abuse and neglect. Nurses must recognize their feelings regarding "proper" discipline and their fear of "getting involved." The feeling that "it's none of my business" must be overcome, and positive attitudes toward the parents and reporting must be promoted. Recognition of legal and moral responsibilities must take precedence over personal feelings. Through professional involvement, nurses can contribute to the primary prevention of child abuse and neglect.


Charge disproportionation-induced multiferroics and electric field control of magnetism in a 2D MXene - Mo2NCl2.
Two-dimensional (2D) magnetoelectric multiferroic materials with the coexistence of magnetization and ferroelectric polarization hold potential for application for the development of next-generation nano-memory devices. However, intrinsic 2D multiferroics with a high critical temperature and strong magnetoelectric coupling are still rare to date. Here, we propose a novel mechanism of 2D monolayer multiferroicity. Based on density functional theory (DFT), we predicted that in a Mo2NCl2 monolayer, the non-equilibrium charge disproportionation of Mo ions will induce an out-of-plane electric polarization, making this material a 2D monolayer multiferroic material. More importantly, the magnetic critical temperature is calculated to be ∼168 K, which is larger than those of the recently reported 2D multiferroic and ferromagnetic systems. Our findings also provide a promising platform to control the magnetic properties and electric behavior in 2D multiferroics using an external electric field.


Upstream stimulating factors: highly versatile stress-responsive transcription factors.
Upstream stimulating factors (USF), USF-1 and USF-2, are members of the eucaryotic evolutionary conserved basic-Helix-Loop-Helix-Leucine Zipper transcription factor family. They interact with high affinity to cognate E-box regulatory elements (CANNTG), which are largely represented across the whole genome in eucaryotes. The ubiquitously expressed USF-transcription factors participate in distinct transcriptional processes, mediating recruitment of chromatin remodelling enzymes and interacting with co-activators and members of the transcription pre-initiation complex. Results obtained from both cell lines and knock-out mice indicates that USF factors are key regulators of a wide number of gene regulation networks, including the stress and immune responses, cell cycle and proliferation, lipid and glucid metabolism, and in melanocytes USF-1 has been implicated as a key UV-activated regulator of genes associated with pigmentation. This review will focus on general characteristics of the USF-transcription factors and their place in some regulatory networks.


Evaluation of salivary nitric oxide levels in oral mucosal diseases: A controlled clinical trial.
Lichen planus is a common dermatologic disease to manifest in the oral cavity. Recurrent aphthous ulcers are the most common ulcers of the oral cavity causing discomfort to the patients. These two diseases have different clinical manifestations which require appropriate treatment after correct diagnosis. Though numerous etiological factors have been proposed for these diseases, their true etio-pathogenesis is not yet established and therefore all therapies are palliative and none is effective universally. In light of this, the role of nitric oxide as a mediator in the etio-pathogenesis of these diseases was considered. The present study was undertaken to note the salivary nitric oxide levels as measured through its product nitrite in oral mucosal diseases like lichen planus and recurrent aphthous ulcers and also to ascertain whether salivary nitric oxide level has a role to play as a pathophysiological mediator in these diseases


Systematic evaluation of characteristics of the membrane-based fed-batch shake flask.
The initial part of process development involves extensive screening programs to identify optimal biological systems and cultivation conditions. For a successful scale-up, the operation mode on screening and production scale must be as close as possible. To enable screening under fed-batch conditions, the membrane-based fed-batch shake flask was developed. It is a shake flask mounted with a central feed reservoir with an integrated rotating membrane tip for a controlled substrate release. Building on the previously provided proof of principle for this tool, this work extends its application by constructive modifications and improved methodology to ensure reproducible performance. The previously limited operation window was expanded by a systematic analysis of reservoir set-up variations for cultivations with the fast-growing organism Escherichia coli. Modifying the initial glucose concentration in the reservoir as well as interchanging the built-in membrane, resulted in glucose release rates and oxygen transfer rate levels during the fed-batch phase varying up to a factor of five. The range of utilizable membranes was extended from dialysis membranes to porous microfiltration membranes with the design of an appropriate membrane tip. The alteration of the membrane area, molecular weight cut-off and liquid volume in the reservoir offered additional parameters to fine-tune the duration of the initial batch phase, the oxygen transfer rate level of the fed-batch phase and the duration of feeding. It was shown that a homogeneous composition of the reservoir without a concentration gradient is ensured up to an initial glucose concentration of 750 g/L. Finally, the experimental validity of fed-batch shake flask cultivations was verified with comparable results obtained in a parallel fed-batch cultivation in a laboratory-scale stirred tank reactor. The membrane-based fed-batch shake flask is a reliable tool for small-scale screening under fed-batch conditions filling the gap between microtiter plates and scaled-down stirred tank reactors. The implemented reservoir system offers various set-up possibilities, which provide a wide range of process settings for diverse biological systems. As a screening tool, it accurately reflects the cultivation conditions in a fed-batch stirred tank reactor and enables a more efficient bioprocess development.


Toward understanding amino acid adsorption at metallic interfaces: a density functional theory study.
In examining adsorption of a few selected single amino acids on Au and Pd cluster models by density functional theory calculations, we have shown that specific side-chain binding affinity to the surface may occur because of a combination of effects, including charge transfer. Larger binding was calculated at the Pd interface. In addition, the interplay between amino acid solvation and adsorption at the interface was considered from first principles. This analysis serves as the first step toward gaining a more accurate understanding of specific interactions at the interface of biological-metal nanostructures than has been attempted in the past.


Progress towards an HIV vaccine based on recombinant bacillus Calmette-Guérin: failures and challenges.
The need for an affordable, safe and effective HIV vaccine has never been greater. As the immunogenicity of all the vaccine vectors being evaluated currently in human populations is limited, novel vaccine strategies are needed to stimulate the innate immune system, to generate high levels of neutralizing antibodies and to induce strong cell-mediated and mucosal immunity. There is strong evidence for a role for cytotoxic T lymphocytes in the containment of HIV replication. Several vaccine approaches have been tested to elicit anti-HIV cytotoxic T-lymphocyte responses. One promising approach is Bacillus Calmette-Guérin (BCG) as a bacterial live recombinant vaccine vehicle. BCG has a long record of safety in humans and is able to induce long-lasting immunity. In this review, we describe the limitations and challenges of developing a recombinant BCG-based HIV vaccine. We also emphasize possible approaches for overcoming the plasmid instability in vivo and the low levels of gene expression and immunogenicity induction. Today, projects all over the world are focused on the development of an AIDS vaccine. Overcoming the remaining scientific, logistical and financial hurdles to the development of an effective HIV vaccine will require real imagination and firm commitment from all stakeholders.


Adverse outcomes of (post-)term births and differences in professional organisational contexts: an integral descriptive system approach.
The development and application of an integral descriptive model for monitoring and evaluation of patient flows and adverse outcomes of childbirth related to distinct categories of professional organisational contexts. After categorisation of the individual professional contexts in which deliveries take place, the resulting framework has been superimposed on the 2002-2010 database of the Netherlands Perinatal Registry. All Dutch hospitals and almost all first-line midwife practices recorded 1,469,955 (post-)term births from which only the patients with a spontaneous onset of labour (n=1,120,508) were included in a study on the quality of obstetric care outside office hours. For the performance of professional organisational contexts the difference in relative incidence of perinatal death or Apgar score <7 between the related patient groups and the reference patient groups has been used. These differences have been expressed as risk ratio (RR) with a 95% CI. Only the group of patients who started labour spontaneously under the supervision of the first-line midwife shows a proportional distribution over the parts of the day. In all other groups the distribution of patients is disproportional. The perinatal mortality rate declines by about 30% in the successive periods. This decline concerns mainly those patients who gave birth outside office hours. The complexity and the dynamics of the obstetric care system make it virtually impossible to demonstrate fixed patterns in the relationships between the separate contextual factors and the (adverse) outcomes of births. To generate useful knowledge, it is necessary to evaluate changes in the obstetric care system periodically and systematically. Thus, the longitudinal application of the model demonstrated that the differences in perinatal mortality rate between the parts of the day have disappeared in recent years.


Purine nucleoside antimetabolites in development for the treatment of cancer.
Purine nucleoside analogs are an important class of drugs that are used in the treatment of cancer. Five purine analogs have been approved by the FDA (mercaptopurine, thioguanine, fludarabine monophosphate, deoxycoformycin and cladribine) and four compounds are currently being evaluated clinically (clofarabine, immucillin-H, nelarabine and 8-chloroadenosine). In addition, two gene therapy approaches are being evaluated that are based on the selective activation of purine nucleoside analogs (ganciclovir, fludarabine monophosphate and others) in tumor cells. Even though nucleoside analogs have been extensively evaluated over the last 50 years, the development of these new compounds demonstrates that there is still much promise in identifying new anticancer drugs from this class of compounds.


Effect of the Buffer on the Buildup and Stability of Tannic Acid/Collagen Multilayer Films Applied as Antibacterial Coatings.
The deposition of polyelectrolyte multilayers, obtained by the layer-by-layer (LbL) method, is a well-established technology to design biocompatible and antibacterial coatings aimed at preventing implant-associated infections. Several types of LbL films have been reported to exhibit antiadhesive and/or antibacterial (contact-killing or release-killing) properties governed not only by the incorporated compounds but also by their buildup conditions or their postbuildup treatments. Tannic acid (TA), a natural polyphenol, is known to inhibit the growth of several bacterial strains. In this work, we developed TA/collagen (TA/COL) LbL films built in acetate or citrate buffers at pH 4. Surprisingly, the used buffer impacts not only the physicochemical but also the antibacterial properties of the films. When incubated in physiological conditions, both types of TA/COL films released almost the same amount of TA depending on the last layer and showed an antibacterial effect against Staphylococcus aureus only for citrate-built films. Because of their granular topography, TA/COL citrate films exhibited an efficient release-killing effect with no cytotoxicity toward human gingival fibroblasts. Emphasis is put on a comprehensive evaluation of the physicochemical parameters driving the buildup and the antibacterial property of citrate films. Specifically, complexation strengths between TA and COL are different in the presence of the two buffers affecting the LbL deposition. This work constitutes an important step toward the use of polyphenols as an antibacterial agent when incorporated in LbL films.


A case of warm autoimmune haemolytic anaemia with intravascular haemolysis: a rare presentation.
Autoimmune haemolytic anaemia (AIHA) is a rare (prevalence 0.2 -1.0 per 100,000 population) but a potentially fatal condition. Diagnosis of AIHA is based mainly on Direct Agglutination Test (DAT/Coomb's test). AIHA is classified into warm (optimal autoantibody reactivity at 37°C) and cold (optimal autoantibody reactivity at 4°C) types. Based on the presence or absence of an underlying cause, it is divided into primary (idiopathic) and secondary (secondary to lympho proliferative disorders, autoimmune disease, drugs and non-haematological malignancies). Warm type accounts for about 70% of cases with AIHA. It can occur at any age, but mostly after 40 years. Haemolysis in warm AIHA is mediated by IgG alone or IgG with complement. Once immunoglobulins are bound, the red cells are taken up by the macrophages of the reticulo-endothelial system which has receptors for the Fc fragments of the immunoglobulins. As the red cells are haemolysed extravascularly this occurs mainly in the spleen. Intravascular haemolysis is unusual in warm AIHA, although it can occur rarely, as warm autoantibodies can fix complements. We report severe warm AIHA in an adult with evidence of intravascular haemolysis, which is an unusual presentation.


Sensitivity to food additives, vaso-active amines and salicylates: a review of the evidence.
Although there is considerable literature pertaining to IgE and non IgE-mediated food allergy, there is a paucity of information on non-immune mediated reactions to foods, other than metabolic disorders such as lactose intolerance. Food additives and naturally occurring 'food chemicals' have long been reported as having the potential to provoke symptoms in those who are more sensitive to their effects. Diets low in 'food chemicals' gained prominence in the 1970s and 1980s, and their popularity remains, although the evidence of their efficacy is very limited. This review focuses on the available evidence for the role and likely adverse effects of both added and natural 'food chemicals' including benzoate, sulphite, monosodium glutamate, vaso-active or biogenic amines and salicylate. Studies assessing the efficacy of the restriction of these substances in the diet have mainly been undertaken in adults, but the paper will also touch on the use of such diets in children. The difficulty of reviewing the available evidence is that few of the studies have been controlled and, for many, considerable time has elapsed since their publication. Meanwhile dietary patterns and habits have changed hugely in the interim, so the conclusions may not be relevant for our current dietary norms. The conclusion of the review is that there may be some benefit in the removal of an additive or a group of foods high in natural food chemicals from the diet for a limited period for certain individuals, providing the diagnostic pathway is followed and the foods are reintroduced back into the diet to assess for the efficacy of removal. However diets involving the removal of multiple additives and food chemicals have the very great potential to lead to nutritional deficiency especially in the paediatric population. Any dietary intervention, whether for the purposes of diagnosis or management of food allergy or food intolerance, should be adapted to the individual's dietary habits and a suitably trained dietitian should ensure nutritional needs are met. Ultimately a healthy diet should be the aim for all patients presenting in the allergy clinic.


Involvement of Porphyromonas gingivalis in the progression of non-alcoholic fatty liver disease.
The risk factors in the progression of nonalcoholic fatty liver disease (NAFLD) have not been fully clarified. Porphyromonas gingivalis (P.g) has been considered to be a confounding risk factor for systemic diseases. We aimed to evaluate the effect of P.g infection on risk of progression to NASH. (1) Serum IgG antibody titers against P.g fimbriae (fimA) in 200 biopsy-proven NAFLD patients were measured by ELISA and compared with histological findings. (2) C57BL/6J mice were fed a control diet (CD) or high-fat diet (HFD) with or without P.g-odontogenic infection and analyzed histologically. Mouse livers were analyzed using CE-TOFMS and LC-TOFMS. (1) A significant correlation between fibrosis progression and antibody titers against P.g possessing fimA type 4 was identified (P = 0.0081). Multivariate analysis identified older age and type 4 P.g-positivity as risk factors for advanced fibrosis. (2) Fibrosis and steatosis were more severe in HFD P.g(+) mice compared with HFD P.g(-) mice. In metabolome analysis, fatty acid metabolism was significantly disrupted with HFD in P.g-infected mouse livers. Monounsaturated/saturated fatty acid ratios were significantly higher in the HFD P.g(+) group than in the HFD P.g(-) group (P < 0.05). Moreover, expression levels of SCD1 and ELOVL6 were significantly reduced. These results suggest that P.g infection is an important risk factor for pathological progression in NAFLD. Increase in the monounsaturated/saturated fatty acid ratio may be an important change that facilitates progression of NAFLD.