Redo-laparoscopy in the management of complications after laparoscopic colorectal surgery: a systematic review and meta-analysis of surgical outcomes.
The laparoscopic approach for colorectal surgery has gradually become widely accepted for the treatment of both benign and malignant diseases thanks to its several advantages over the open approach. However, it is associated with the same potential postoperative complications. Some recent studies have analyzed the potential role of laparoscopy in early diagnosis and management of complications following laparoscopic colorectal surgery. The aim of this systematic review was to investigate the outcomes of redo-laparoscopy (RL) for the management of early postoperative complications following laparoscopic colorectal surgery, focusing on length of stay, morbidity and mortality. A systematic review of the literature was performed according to the preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines through MEDLINE (PubMed), Embase and Google Scholar from January 1990 to December 2019. The main outcomes examined were conversion rate, length of hospital stay, postoperative morbidity and mortality rates. A meta-analysis of all eligible studies was then conducted and forest plots were generated. A total of 19 studies involving 1394 patients who required reoperation after laparoscopic colorectal resection were included. In 539 (38.2%) of these patients, a laparoscopic approach was adopted. The most common indication for returning to the operating theater was anastomotic leakage (64.4% of all redo-surgeries, 67.7% of RL) and the most common type of intervention performed in RL was diverting stoma with or without anastomotic repair/redo (47.1%). Nine studies were included in the pooled analysis. The mean length of stay was significantly shorter in the RL group than in the redo-open one (WMD = - 0.90; 95% CI - 1.04 to - 0.76; Z = - 12,6; p < 0.001). A significantly lower risk of mortality was observed in the RL cohort (OR = - 0.91; 95% CI - 1.58 to - 0.23; Z = - 2.62; p = 0.009). Laparoscopy is a valid and effective approach for the treatment of complications following laparoscopic primary colorectal surgery thanks to it is well-established advantages over the open approach, which remain noticeable even in redo-surgeries.


Extended experience with a non-cytotoxic DNMT1-targeting regimen of decitabine to treat myeloid malignancies.
The nucleoside analogue decitabine can deplete the epigenetic regulator DNA methyltransferase 1 (DNMT1), an effect that occurs, and is saturated at, low concentrations/doses. A reason to pursue this molecular-targeted effect instead of the DNA damage/cytotoxicity produced with high concentrations/doses, is that non-cytotoxic DNMT1-depletion can cytoreduce even p53-null myeloid malignancies while sparing normal haematopoiesis. We thus identified minimum doses of decitabine (0·1-0·2 mg/kg) that deplete DNMT1 without off-target anti-metabolite effects/cytotoxicity, and then administered these well-tolerated doses frequently 1-2X/week to increase S-phase dependent DNMT1-depletion, and used a Myeloid Malignancy Registry to evaluate long-term outcomes in 69 patients treated this way. Consistent with the scientific rationale, treatment was well-tolerated and durable responses were produced (~40%) in genetically heterogeneous disease and the very elderly.


USA300 methicillin-resistant Staphylococcus aureus in Norway.
Moen AEF*, Tannaes TM, Leegaard TM. USA300 methicillin-resistant Staphylococcus aureus (MRSA) in Norway. Community-associated methicillin-resistant S. aureus USA300 is known for its ability to disseminate and colonize and the clone has been detected globally. We studied the most populated area in Norway in search for the strain and for signs of endemic establishment. Strain typing techniques such as spa-, SCCmec- and dru-typing, pulsed-field gel electrophoresis and detection of USA300 molecular markers, lukS/F-PV and arcA, were performed. USA300 has been present in the study area since 2003 and has increased in incidence in parallel with the total MRSA incidence. Most USA300 isolates were found in the community, but isolates were also detected in health care institutions.


Digitized fluorescent RFLP analysis (fRFLP) as a universal method for comparing genomes of culturable dsDNA viruses: application to bacteriophages.
Identification and classification of bacteriophages remains a cumbersome process even with the use of genotypic approaches, due to the lack of genes present in all phages. Restriction fragment length polymorphism analysis (RFLP) of the viral genome is a universal approach, but RFLP fingerprints obtained on agarose gels remain difficult to compare between laboratories. Here we describe the digitization of RFLP of viral genomes by amplification of all restriction fragments - after ligation of adapters - using primers complementary to the adapters only. Since one of the primers is fluorescently labelled, the restriction fragments become visible to a fluorescent capillary electrophoresis system (ABI310) and their lengths can be digitized immediately. The digitized fluorescent RFLP (fRFLP) fingerprint can be stored as an entry in a library. Dendrogram construction of the fRFLP fingerprints obtained for a total of 69 Caudovirales (tailed bacteriophages) showed that genomically and/or serologically closely related phages clustered, whereas host range was not completely in correspondence with genotype. fRFLP might be a tool for quickly establishing the relationship of newly isolated phages to previously isolated ones and for constructing an fRFLP library electronically accessible on the internet, to which fRFLP patterns of new phages can be compared.


Alteration of the N6-methyladenosine methylation landscape in a mouse model of polycystic ovary syndrome.
To explore the N6-methyladenosine (m6A) methylation abnormality of mRNAs and its potential roles in the mouse model of polycystic ovary syndrome (PCOS). The mouse model of PCOS were induced by injecting dehydroepiandrosterone (DHEA), and confirmed by observing the morphological structures of ovarian follicles. Subsequently, m6A-tagged mRNAs were identified via m6A epitranscriptomic microarray and its potential functional pathways were predicted in KEGG database. The expression and modification levels of key mRNAs in the most enriched pathway were evaluated and compared using western blot and methylated RNA immunoprecipitation-quantitative PCR (MeRIP-qPCR). Compared with the control group, 415 hypermethylated and downregulated mRNAs, 8 hypomethylated and upregulated mRNAs, and 14 hypermethylated and upregulated mRNAs were identified in the PCOS group (Fold change ≥ 1.5). Those mRNAs were mainly involved in insulin signaling pathway, type II diabetes mellitus, Fc epsilon RI signaling pathway, inositol phosphate metabolism, and GnRH secretion. In insulin signaling pathway, the expression levels of phosphorylated protein kinase B (p-AKT) were decreased, whereas that of upstream phosphorylated phosphatidylinositol 3-kinase (p-PI3K) were increased in PCOS group. Moreover, skeletal muscle and kidney-enriched inositol polyphosphate 5-phosphatease (SKIP), one of PIP3 phosphatases, was verified to be overexpressed, and Skip mRNAs were hypermethylated in PCOS group. The altered m6A modification of mRNAs might play a critical role in PCOS process. The PI3K/AKT pathway is inhibited in the mouse model of PCOS. Whether it is caused by the m6A modification of Skip mRNAs is worthy of further exploration.


Benzocaine--an unsatisfactory indicator of topical local anaesthetic sensitization for the U.K.
Three thousand consecutive patients undergoing patch tests were tested with a mix of benzocaine, amethocaine hydrochloride and cinchocaine hydrochloride. Eighty-four subjects (2.8%) were found to be allergic to this mix. Forty of these were patch tested with the constituents of the mix: 21 (52.5%) patients were sensitive to amethocaine hydrochloride or cinchocaine hydrochloride or both, but not to benzocaine. We recommend that a non-irritant 'caine mix' containing benzocaine, cinchocaine and amethocaine should be included in the standard patch test series for the U.K.


Molecular characterization of a membrane-bound cGMP dependent protein kinase from the silk moth Bombyx mori.
The cGMP signalling pathway has been suggested to be involved in the signal transduction of various physiological functions in insects; olfaction, antidiuresis and eclosion. However, the cGMP signalling mechanism has remained elusive. We isolated two cDNAs of the cGMP dependent protein kinase, designated BmPKG-Ialpha and BmPKG-Ibeta. The deduced amino acid sequences indicate that both BmPKG-Ialpha and BmPKG-Ibeta appear to consist of an amino terminal region, a cGMP binding domain and a protein kinase domain. Transcripts of BmPKG-Ialpha and BmPKG-Ibeta were detected in various tissues: flight muscles, antennae, midgut, legs, head, thoracic ganglia and Malphighian tubules. Recombinant BmPKG-Ialpha bound to lipid membranes, while BmPKG-Ialpha with a deleted amino terminal region failed to bind to lipid membranes.


Structure and dynamics of plasmalogen model membranes containing cholesterol: a deuterium NMR study.
Deuterium nuclear magnetic resonance (2H-NMR) was used to investigate the structure and dynamics of the sn-2 hydrocarbon chain of semi-synthetical choline and ethanolamine plasmalogen in bilayers containing 0, 30, and 50 mol% cholesterol. The deuterium NMR spectra of the choline plasmalogen yielded well-resolved quadrupolar splittings which could be assigned to the corresponding hydrocarbon chain deuterons. The sn-2 acyl chain was found to adopt a similar conformation as observed in the corresponding diacyl phospholipid, however, the flexibility at the level of the C-2 methylene segment of the plasmalogen was increased. Deuterium NMR spectra of bilayers composed of the ethanolamine plasmalogen yielded quadrupolar splittings of the C-2 segment much larger than those of the corresponding diacyl lipids, suggesting that the sn-2 chain is oriented perpendicular to the membrane surface at all segments. Cholesterol increased the ordering of the choline plasmalogen acyl chain to the same extent as in diacyl lipid bilayers. T1 relaxation time measurements demonstrated only minor dynamical differences between choline plasmalogen and diacyl lipids in model membranes.


Slower rescue of ER homeostasis by the unfolded protein response pathway associated with common variable immunodeficiency.
Common variable immunodeficiency (CVID) is a primary immunodeficiency characterized by hypogammaglobulinemia and recurrent infections. Herein we addressed the role of unfolded protein response (UPR) in the pathogenesis of the disease. Augmented unspliced X-box binding protein 1 (XBP-1) mRNA concurrent with co-localization of IgM and BiP/GRP78 were found in one CVID patient. At confocal microscopy analysis this patient's cells were enlarged and failed to present the typical surface distribution of IgM, which accumulated within an abnormally expanded endoplasmic reticulum. Sequencing did not reveal any mutation on XBP-1, neither on IRE-1alpha that could potentially prevent the splicing to occur. Analysis of spliced XBP-1, IRE-1alpha and BiP messages after LPS or Brefeldin A treatment showed that, unlike healthy controls that respond to these endoplasmic reticulum (ER) stressors by presenting waves of transcription of these three genes, this patient's cells presented lower rates of transcription, not reaching the same level of response of healthy subjects even after 48 h of ER stress. Treatment with DMSO rescued IgM and IgG secretion as well as the expression of spliced XBP-1. Our findings associate diminished splicing of XBP-1 mRNA with accumulation of IgM within the ER and lower rates of chaperone transcription, therefore providing a mechanism to explain the observed hypogammaglobulinemia.


Property Control of Layer-by-Layer Assembled Sulfonated Poly(phenylene oxide) Composite Membrane Based on Nafion for Fuel Cell Applications.
Multilayered composite proton exchange membranes were prepared by LbL method by alternating deposition of poly(diallyl dimethyl ammonium chloride) (PDDA) and highly sulfonated poly(phenylene oxide) (sPPO) onto the surface of Nafion 212. The sulfonated sPPO solution contained polystyrenesulfonic acid copolymer with azide moiety. Thickness of the LbL composite membrane was controlled by using the anionic solution of sPPO containing 0.5 M NaCl. The membranes were crosslinked by using UV to give the mechanical and chemical durability. The crosslinked composite membrane showed decreased methanol permeability with the increasing number of bilayers, and showed increased overall selectivity compared to Nafion film.


Impact of therapeutic hypothermia on bleeding events in adult patients treated with extracorporeal life support peri-cardiac arrest.
Whether therapeutic hypothermia (TH) adds to the risk of bleeding in patients on extracorporeal life support (ECLS) peri-cardiac arrest remains unknown. Single center retrospective study on patients receiving veno-arterial ECLS peri-cardiac arrest ± TH at 32-34 °C (January 2009-December 2015). major bleeding (including intracerebral hemorrhage, ICH) < 72 h of cardiac arrest. Logistic regression and marginal structural models were used to analyze associations with major bleeding. Of 66 patients receiving ECLS, 36 were treated with TH. Major bleeding occurred in 14 patients (39%) treated with ECLS+TH and in 17 patients (57%) with ECLS alone. ICH was reported in 3 (8%) and one patient (3%), respectively. There was no difference in mortality, but lung injury occurred more often in ECLS+TH. A platelet count <60 × 109/L but not TH was associated with major bleeding (including ICH). The estimated causal risk ratio of TH on the occurrence of major bleeding (including ICH) at 72 h post cardiac arrest was 0.95 (95%CI 0.62-1.45). Bleeding complications were common in our study. However, TH (32-34 °C) was not associated with an increased risk of major bleeding in patients on ECLS peri-cardiac arrest.


Molecular characterization of the largest subunit of Plasmodium falciparum RNA polymerase I.
Plasmodium species possess developmentally regulated ribosomal RNA (rRNA) genes. This report describes the expression and gene structure of the largest subunit of P. falciparum RNA polymerase I (RNAPI), which is responsible for the synthesis of rRNA. The RNAPI largest subunit gene was present as a single copy gene on chromosome 9. Three exons encode the 2910-amino acid RNAPI polypeptide (340 140 Da). A comparison of Plasmodium, Trypanosoma, and Saccharomyces cerevisiae nuclear RNAP largest subunits identified conserved amino acid positions and class-specific amino acid positions. Novel amino acid insertions were found between RNAPI conserved regions A and B (region A'), D and DE1 (region D'), DE2 and E (region DE2'), and F and G (region F'). Leucine zipper domains were found within regions D', DE2, and DE2'. A novel serine-rich repeat domain, a domain with homology to the C-terminal domain of eukaryotic upstream binding factor (UBF), and 4 highly conserved casein kinase II (CKII) Ser/Thr phosphorylation motifs were found within a 127-amino acid sub-region of enlarged region F'. The novel RNAPI serine-rich repeat contained a conserved motif, Ser-X3-Ser, which was also identified in the serine-rich repeat domains of the P. falciparum RNAPII and RNAPIII largest subunits, as well as within a highly homologous serine-rich repeat from trophozoite antigen R45. The results of this molecular analysis indicate that phosphorylation and dephosphorylation mechanisms regulate the activity of P. falciparum RNAPI.


Pharmacokinetics and bioequivalence evaluation of risperidone in healthy male subjects with different CYP2D6 genotypes.
The aim of this study was to evaluate the bioequivalence of risperidone in healthy male subjects representing different CYP2D6 genotypes with respect to risperidone, 9-hydroxyrisperidone (9-OH-risperidone), and active moiety. A total of 506 Korean subjects were genotyped for CYP2D6*10 by means of allele-specific polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Based on the genotype analysis, 24 subjects, 7 homozygous for CYP2D6*1, 10 for *10, and 7 heterozygous for *10, were recruited and received a single oral dose of 2 mg risperidone tablet in this study. Serum concentrations of risperidone and 9-OHrisperidone up to 48 h were simultaneously determined. There were no significant differences of the active moiety, risperidone, and 9-OH-risperidone between the two preparations in AUC0-proportinal to, and Cmax. The 90% confidence intervals (CIs) for the ratio of means of the log-transformed AUC0-proportional to. and Cmax for the active moiety, risperidone, and 9-OH-risperidone were all within the bioequivalence acceptance criteria of 0.80-1.25. The CYP2D6*10 allele particularly was associated with higher serum concentrations of risperidone and the risperidone/9-OH-risperidone ratio compared with the CYP2D6*1 allele. The results demonstrate that the two preparations of risperidone are bioequivalent and it can be assumed that they are therapeutically equivalent and exchangeable in clinical practice. Furthermore, the pharmacokinetic parameters of risperidone and the risperidone/9-OH-risperidone ratio are highly dependent on the CYP2D6 genotypes.


Two types of synaptic glomeruli and their distribution in laminae I-III of the rat spinal cord.
Systematic examination of photomontages revealed two types of synaptic glomeruli in laminae II-III. Type I glomeruli have a dark small central (C) terminal of indented contour with closely packed spherical vesicles of variable diameter and few mitochondria. Among the peripheral terminals there are dendritic spines and a few presynaptic dendritic spines (V1 terminals) and axon endings rich in discoid vesicles (V2). These glomeruli occur in groups which are particularly evident in parasagittal sections in which successive C terminals are connected by narrower portions or dark unmyelinated profiles. Type II glomeruli have an electron-lucent and large C terminal of regular contour with less packed synaptic vesicles of more uniform diameter, more mitochondria, and sometimes neurofilaments. Presynaptic dendrites are fewer and axon endings more numerous. C terminals in type II glomeruli are fusiform in longitudinal section, rarely occurring in groups. Lamina I is virtually devoid of glomeruli. Within lamina II, glomeruli are rare in the dorsalmost 20-micrometers band and abundant in the immediately ventral 20-micrometers band in which type I glomeruli are prevalent (approximately 79%). In ventral lamina II, type II glomeruli predominate (66%), being practically exclusive in lamina III where most contain neurofilaments. Considering the distribution of terminations of primary afferents, it is suggested that type I C terminals originated from unmyelinated primary afferents, type II C terminals without neurofilaments from direct myelinated fibers, the those with neurofilaments from recurrent large fibers. The distinct numbers of presynaptic dendritic and axonal endings suggest different modulatory mechanisms functioning in the two types and in ventral vs. dorsal areas of this region.


Quaternized agricultural by-products as anion exchange resins.
The objectives of this study were the chemical modification of readily available, low-cost agricultural by-products to anion exchange resins and the selection of the best modified by-product for further use in anion removal. Resins were prepared through the quaternization of a series of 12 agricultural by-products with N-(3-chloro-2-hydroxypropyl) trimethylammonium chloride (CHMAC). Phosphate ion adsorption assays were conducted at pH 7 in order to compare adsorption properties among the by-products. Quaternized corn stover showed the highest phosphorus adsorption at 0.66 mmole/g. Since corn stover exhibited the best uptake of phosphate ion, it was compared to a commercially available, cellulose-based anion exchange resin. Additionally, adsorption capacities of quaternized corn stover for arsenate, chromate, and selenate were evaluated and adsorption efficiencies were determined in simulated wastewater samples. Our results indicate that modified corn stover demonstrates good adsorption uptake for arsenate and selenate and especially for chromate.


Multiplexed quantitative proteomics provides mechanistic cues for malaria severity and complexity.
Management of severe malaria remains a critical global challenge. In this study, using a multiplexed quantitative proteomics pipeline we systematically investigated the plasma proteome alterations in non-severe and severe malaria patients. We identified a few parasite proteins in severe malaria patients, which could be promising from a diagnostic perspective. Further, from host proteome analysis we observed substantial modulations in many crucial physiological pathways, including lipid metabolism, cytokine signaling, complement, and coagulation cascades in severe malaria. We propose that severe manifestations of malaria are possibly underpinned by modulations of the host physiology and defense machinery, which is evidently reflected in the plasma proteome alterations. Importantly, we identified multiple blood markers that can effectively define different complications of severe falciparum malaria, including cerebral syndromes and severe anemia. The ability of our identified blood markers to distinguish different severe complications of malaria may aid in developing new clinical tests for monitoring malaria severity.


Reliability and concurrent validity of bathroom weighing scale and sphygmomanometer in quantifying magnitude of digital postero-anterior spinal pressure.
The main objective of this study was to determine the reliability and concurrent validity of using Modified Sphygmomanometer (sphyg) and body weighing scale to determine the magnitude of pressure and body weight change during digital spinal examination. One hundred and twenty apparently healthy subjects were recruited. Weighing scale was used to measure the body weight of the investigator while Mercury sphygmomanometer was used to measure the pressure exerted. There was test re-test interval of 2 weeks. Data were analysed by descriptive statistic and Pearson product moment correlation. The result showed that there was significant correlation between the weight change (WC) values obtained on the weighing scale on day 1 and 2 (r=0.91). Also, there was significant correlation between Pressure obtained in the sphygmomanometer on day 1 and 2 (r=0.94). Similarly, there was significant correlation between the WC and Pressure Change (PC), (r=-0.49). The participant's age, height, weight and BMI contributed significantly as predictors for both BWC and PC change respectively. This study concluded that weighing scale and Mercury sphygmomanometer are reliable and valid to determine the magnitude of pressure applied during DPAP.


Intracortical bone remodeling variation shows strong genetic effects.
Intracortical microstructure influences crack propagation and arrest within bone cortex. Genetic variation in intracortical remodeling may contribute to mechanical integrity and, therefore, fracture risk. Our aim was to determine the degree to which normal population-level variation in intracortical microstructure is due to genetic variation. We examined right femurs from 101 baboons (74 females, 27 males; aged 7-33 years) from a single, extended pedigree to determine osteon number, osteon area (On.Ar), haversian canal area, osteon population density, percent osteonal bone (%On.B), wall thickness (W.Th), and cortical porosity (Ct.Po). Through evaluation of the covariance in intracortical properties between pairs of relatives, we quantified the contribution of additive genetic effects (heritability [h (2)]) to variation in these traits using a variance decomposition approach. Significant age and sex effects account for 9 % (Ct.Po) to 21 % (W.Th) of intracortical microstructural variation. After accounting for age and sex, significant genetic effects are evident for On.Ar (h (2) = 0.79, p = 0.002), %On.B (h (2) = 0.82, p = 0.003), and W.Th (h (2) = 0.61, p = 0.013), indicating that 61-82 % of the residual variation (after accounting for age and sex effects) is due to additive genetic effects. This corresponds to 48-75 % of the total phenotypic variance. Our results demonstrate that normal, population-level variation in cortical microstructure is significantly influenced by genes. As a critical mediator of crack behavior in bone cortex, intracortical microstructural variation provides another mechanism through which genetic variation may affect fracture risk.


Electrochemical Reductive Functionalization of Alkenes with Deuterochloroform as a One-Carbon Deuteration Block.
Deuterochloroform is the most common organic deuterated solvent. However, the synthetic chemistry of deuterochloroform has been marginally explored because of its intrinsic limitations, including elimination and scrambling. In this work, an electrochemical protocol was developed to use deuterochloroform as a one-carbon deuteration block in the cyclopropanation and hydrochloromethylation of alkenes. By employing different reaction conditions, the chemoselectivity could be regulated to give divergent products. Typically, deuterium incorporation above 93% to 99% could be achieved.


Poorly differentiated sarcoma of the maxillary sinus: a histopathology dilemma of a rare tumor.
Sarcomas are a rare heterogeneous group of neoplasms of mesenchymal origin. In the redistribution of all head and neck malignancies, sarcomas are represented by only 1%. Herein, we report a case of a 66-year-old patient with right maxillary sinus sarcoma that spread through the ostiomeatal complex, infiltrated the septum, all ethmoid cells, frontal sinus, involved the entire right nasal cavity and penetrated to the nasopharynx. Patient was treated with neoadjuvant chemotherapy, surgery and adjuvant radiotherapy. The histopathology indicated poorly differentiated sarcoma with elements of Ewing's sarcoma, but also with elements consistent with osteosarcoma. Molecular pathological analysis excluded Ewing's sarcoma. Samples were also sent for review to the other Pathology Clinics. They suggested poorly differentiated high-grade pleomorphic sarcoma with elements of osteosarcoma. The accurate diagnosis of the head and neck sarcoma type can be a histopathology dilemma posing a great challenge in the choice of therapeutic approach, and thus the treatment outcome.


Interspecific competition between the principal larval parasitoids of the pine sawfly, Neodiprion sertifer (Geoff.) (Hym.: Diprionidae).
Interspecific competition between the 3 principal larval parasitoids of the pine sawfly, Neodiprion sertifer, is of common occurrence when total larval parasitism, and hence multiple attack rates, are high. At the intrinsic level, the ectoparasitoid Exenterus abruptorius is superior to the 2 endoparasitoids, Lophyroplectus luteator and Lamachus eques, respectively, whereas L. luteator is superior to L. eques only. During mass outbreaks of the host competition between Exenterus and Lophyroplectus is most intense, whilst Lamachus fails to build up in the presence of its 2 competitors (Fig. 1). However, due to its superior host finding ability, Lamachus dominates in areas of low host densities where the 2 other species are less efficient (Table 1). L. luteator frequently sustains heavy losses in direct competition with E. abruptorius, but these can be largely compensated in the next generation because its fertility is about 5-times higher than that of Exenterus.


Properties of a novel glucose-enhanced beta-glucosidase purified from Streptomyces sp. (ATCC 11238).
An inducible intracellular beta-glucosidase (EC 126.96.36.199) from Streptomyces sp. QM-B814 (ATCC 11238) has been purified and characterized. The purified polypeptide is monomeric with a relative molecular mass of 62 kDa by SDS-PAGE and 42 kDa by size-exclusion chromatography; its isoelectric point is 4.2. The difference in the molecular mass values can be attributed to the glycosylated nature of the protein. The purified enzyme has a pH optimum of 6.0-6.5. The temperature optimum for activity is 50 degrees C; at this temperature the enzyme is stable for 1 h. The enzyme hydrolyzes mainly aryl-beta-glucosides but also presents significant activity against beta-linked disaccharides and maltose. The enzyme displays an unusual kinetic behavior and biphasic Lineweaver-Burk and Eadie-Hofstee plots for p-nitrophenyl-beta-D-glucoside and cellobiose were obtained. The enzyme presents beta-glycosyltransferase activity and an exoglycosidase-type action on cellodextrins. It is inhibited by delta-gluconolactone (Ki 0.44 mM) but, remarkably, glucose in the range 25-200 mM enhances the rate of p-nitrophenyl-beta-D-glucoside hydrolysis.


Secondary polycythaemia associated with unilateral renal cystic disease.
Unilateral renal cystic disease (URCD) is a rare condition, with pathological features indistinguishable from autosomal dominant polycystic kidney disease (ADPKD). In contrast to this condition, however, URCD is not inherited, is not associated with progressive deterioration in renal function, and is unilateral. We present a case of URCD associated with polycythaemia, which showed resolution following nephrectomy. Secondary polycythaemia has not previously been reported in cases of URCD, but may be hypothesised to be a result of excess erythropoietin production.


Feed frame: The last processing step before the tablet compaction in pharmaceutical manufacturing.
The feed frame is a force-feeding device used in the die filling process. The die filling process is crucial within pharmaceutical manufacturing to guarantee the critical quality attributes of the tablets. In recent years, interest in this unit has increased because it can affect the properties of the powder blend and tablets, and because of the success in real time monitoring of powder blend uniformity potential for Process Analytical Technology as described in this review. The review focuses on the recent advances in understanding the powder flow behavior inside the feed frame and how the residence time distribution of the powder within the feed frame is affected by the operating conditions and design parameters. Furthermore, this review also highlights the effect of the paddle wheel design and feed frame process parameters on the tablet weight, the principal variable for measuring die filling performance.


Correlation between classification of human urothelial cell lines and HLA-A,B,C expression.
Quantitative changes in major histocompatibility class I antigen expression in tumour cells are believed to affect the host immune response against the tumour. In tumourigenic (TGrIII) human urothelial cell lines the apparent loss of polymorphic HLA-A,B epitopes has previously been demonstrated. In the present study, 3 non-tumourigenic (TGrII) and 6 tumourigenic (TGrIII) human urothelial cell lines have been investigated for their quantitative expression of monomorphic HLA-A,B,C and B2-microglobulin. Evidence is provided that an inverse correlation exists between tumourigenicity and HLA-A,B,C and B2-microglobulin expression. Furthermore, treatment of the cells with neuraminidase partly restored the expression of monomorphic HLA-A,B,C suggesting that at least some of the observed quantitative differences could be due to masking of the membrane bound HLA antigens by sialic acid-containing glycoconjugates.


The evolution of Negative Pressure Wound Therapy [NPWT1: a review of science, available devices, and evolution in the usage of NPWT.
Negative Pressure Wound Therapy (NPWT) is an important clinical tool with multiple published studies reporting more rapid formation of granulation tissue and a decreased time for wound healing.Since the induction of NPWT systems in 1995, there has been wide usage of the device for treatment of wounds. However, the past 18 years has also witnessed a significant shift in the way NPWT is utilized. Meeting the demands and wound care needs of an ever-growing population has resulted in several other companies releasing their versions of the NWPT device. Although there is a significant body of literature supporting the science and clinical advantages of the KCI system, the newer systems differ in terms of suction devices, connecting tubing, and wound interface materials. It cannot be assumed that the newly developed devices have the same science and clinical advantages as the KCI System. The purpose of this article is to review the science behind NPWT, assess and compare the currently available devices, and examine the scientific literature supporting the efficiency and application of NPWT in wound reconstruction.


Regional cerebral blood flow correlates of visuospatial tasks in Alzheimer's disease.
This study investigated the role of visuospatial tasks in identifying cognitive decline in patients with Alzheimer's disease (AD), by correlating neuropsychological performance with cerebral perfusion measures. There were 157 participants: 29 neurologically healthy controls (age: 70.3 +/- 6.6, MMSE > or = 27), 86 patients with mild AD (age: 69.18 +/- 8.28, MMSE > or = 21) and 42 patients moderate/severe AD (age: 68.86 +/- 10.69, MMSE 8-20). Single Photon Emission Computerized Tomography (SPECT) was used to derive regional perfusion ratios, and correlated using partial least squares (PLS) with neuropsychological test scores from the Benton Line Orientation (BLO) and the Rey-Osterrieth Complex Figure (RO). Cross-sectional analysis demonstrated that mean scores differed in accordance with disease status: control group (BLO 25.5, RO 33.3); mild AD (BLO 20.1, RO 25.5); moderate/severe AD (BLO 10.7, RO 16). Correlations were observed between BLO/RO and right parietal SPECT regions in the AD groups. Visuospatial performance, often undersampled in cognitive batteries for AD, is clearly impaired even in mild AD and correlates with functional deficits as indexed by cerebral perfusion ratios on SPECT implicating right hemisphere circuits. Furthermore, PLS reveals that usual spatial tasks probe a distributed brain network in both hemispheres including many areas targeted by early AD pathology.


CD86 molecule is a specific marker for canine monocyte-derived dendritic cells.
In this study, canine monocyte-derived dendritic cells (cMo-DC) were produced in presence of canine GM-CSF (cGM-CSF) and canine IL-4 (cIL-4), and they were characterized by their dendritic morphology, MLR functionality and phenotype. We noticed that cMo-DC were labelled with three anti-human CD86 (FUN-1, BU63 and IT2.2 clones), whereas resting and activated lymphocytes or monocytes were not stained. CD86 expression was induced by cIL-4 and was up-regulated during the differentiation of the cMo-DC, with a maximum at day 7. Furthermore, cMo-DC were very potent even in low numbers as stimulator cells in allogeneic MLR, and BU63 mAb was able to completely block the cMo-DC-induced proliferation in MLR. We also observed that cMo-DC highly expressed MHC Class II and CD32, but we failed to determine their maturation state since the lack of commercially available canine markers. Moreover, cMo-DC contained cytoplasmic periodic microstructures, potentially new ultrastructural markers of canine DC recently described. In conclusion, this work demonstrates that the CD86 costimulatory marker is now usable for a better characterization of in vitro canine DC.


Sensorimotor transformation via sparse coding.
Sensorimotor transformation is indispensable to the accurate motion of the human body in daily life. For instance, when we grasp an object, the distance from our hands to an object needs to be calculated by integrating multisensory inputs, and our motor system needs to appropriately activate the arm and hand muscles to minimize the distance. The sensorimotor transformation is implemented in our neural systems, and recent advances in measurement techniques have revealed an important property of neural systems: a small percentage of neurons exhibits extensive activity while a large percentage shows little activity, i.e., sparse coding. However, we do not yet know the functional role of sparse coding in sensorimotor transformation. In this paper, I show that sparse coding enables complete and robust learning in sensorimotor transformation. In general, if a neural network is trained to maximize the performance on training data, the network shows poor performance on test data. Nevertheless, sparse coding renders compatible the performance of the network on both training and test data. Furthermore, sparse coding can reproduce reported neural activities. Thus, I conclude that sparse coding is necessary and a biologically plausible factor in sensorimotor transformation.


Profiles of purine biosynthesis, salvage and degradation in disks of potato (Solanum tuberosum L.) tubers.
To find general metabolic profiles of purine ribo- and deoxyribonucleotides in potato (Solanum tuberosum L.) plants, we looked at the in situ metabolic fate of various (14)C-labelled precursors in disks from growing potato tubers. The activities of key enzymes in potato tuber extracts were also studied. Of the precursors for the intermediates in de novo purine biosynthesis, [(14)C]formate, [2-(14)C]glycine and [2-(14)C]5-aminoimidazole-4-carboxyamide ribonucleoside were metabolised to purine nucleotides and were incorporated into nucleic acids. The rates of uptake of purine ribo- and deoxyribonucleosides by the disks were in the following order: deoxyadenosine > adenosine > adenine > guanine > guanosine > deoxyguanosine > inosine > hypoxanthine > xanthine > xanthosine. The purine ribonucleosides, adenosine and guanosine, were salvaged exclusively to nucleotides, by adenosine kinase (EC 126.96.36.199) and inosine/guanosine kinase (EC 22.214.171.124) and non-specific nucleoside phosphotransferase (EC 220.127.116.11). Inosine was also salvaged by inosine/guanosine kinase, but to a lesser extent. In contrast, no xanthosine was salvaged. Deoxyadenosine and deoxyguanosine, was efficiently salvaged by deoxyadenosine kinase (EC 188.8.131.52) and deoxyguanosine kinase (EC 126.96.36.199) and/or non-specific nucleoside phosphotransferase (EC 184.108.40.206). Of the purine bases, adenine, guanine and hypoxanthine but not xanthine were salvaged for nucleotide synthesis. Since purine nucleoside phosphorylase (EC 126.96.36.199) activity was not detected, adenine phosphoribosyltransferase (EC 126.96.36.199) and hypoxanthine/guanine phosphoribosyltransferase (EC 18.104.22.168) seem to play the major role in salvage of adenine, guanine and hypoxanthine. Xanthine was catabolised by the oxidative purine degradation pathway via allantoin. Activity of the purine-metabolising enzymes observed in other organisms, such as purine nucleoside phosphorylase (EC 126.96.36.199), xanthine phosphoribosyltransferase (EC 220.127.116.11), adenine deaminase (EC 188.8.131.52), adenosine deaminase (EC 184.108.40.206) and guanine deaminase (EC 22.214.171.124), were not detected in potato tuber extracts. These results suggest that the major catabolic pathways of adenine and guanine nucleotides are AMP --> IMP --> inosine --> hypoxanthine --> xanthine and GMP --> guanosine --> xanthosine --> xanthine pathways, respectively. Catabolites before xanthosine and xanthine can be utilised in salvage pathways for nucleotide biosynthesis.


Determining the selectivity of a tetra-phosphorylated biomimetic peptide towards uranium in the presence of competing cations through the simultaneous coupling of HILIC to ESI-MS and ICP-MS.
A cyclic tetra-phosphorylated biomimetic peptide (pS1368) has been proposed as a promising starting structure to design a decorporating agent of uranyl (UO22+) due to its affinity being similar to that of osteopontin (OPN), a target UO22+ protein in vivo. The determination of this peptide's selectivity towards UO22+ in the presence of competing endogenous elements is also crucial to validate this hypothesis. In this context, the selectivity of pS1368 towards UO22+ in the presence of Ca2+, Cu2+ and Zn2+ was determined by applying the simultaneous coupling of hydrophilic interaction chromatography (HILIC) to electrospray ionization (ESI-MS) and inductively coupled plasma (ICP-MS) mass spectrometry. Sr2+ was used as Ca2+ simulant, providing less challenging ICP-MS measurements. The separation of the complexes by HILIC was first set up. The selectivity of pS1368 towards UO22+ was determined in the presence of Sr2+, by adding several proportions of the latter to UO2(pS1368). UO22+ was not displaced from UO2(pS1368) even in the presence of a ten-fold excess of Sr2+. The same approach has been undertaken to demonstrate the selectivity of pS1368 towards UO22+ in the presence of Cu2+, Zn2+ and Sr2+ as competing endogenous cations. Hence, we showed that pS1368 was selective towards UO22+ in the presence of Sr2+, but also in the presence of Cu2+ and Zn2+. This study highlights the performance of HILIC-ESI-MS/ICP-MS simultaneous coupling to assess the potential of molecules as decorporating agents of UO22+.


The gene encoding for the novel transacting factor proopiomelanocortin corticotropin-releasing hormone responsive element binding protein 1 (PCRH-REB-1) is constitutively expressed in rat pituitary and in discrete brain regions containing CRH or CRH receptors: pathophysiological implications.
The nucleotide sequence CTGTGCGCGCAG is a core corticotropin-releasing hormone (CRH) responsive element in the proopiomelanocortin (POMC) promoter that gives 5-7 fold stimulation of POMC transcription by CRH. This region binds a novel transacting factor, proopiomelanocortin corticotropin-releasing hormone responsive element binding protein 1 (PCRH-REB-1), recently cloned from CRH-stimulated pituitary cells. We conducted in situ hybridization histochemistry experiments, using a species-specific, antisense, 35S-labeled PCRH-REB-1 riboprobe, to determine the pattern of PCRH-REB-1 gene expression in the brain, and showed that PCRH-REB-1 mRNA is localized not only in pituitary, but it is also found in discrete brain regions such as septal nuclei, hypothalamus, cortex, hippocampus, amygdala, and cerebellum. Those regions contain CRH or CRH receptors, and respond to CRH. We hypothesize that PCRH-REB-1 may be a marker of cellular responsiveness to CRH in the brain and pituitary, and might therefore be useful in the evaluation of CRH function and in the identification of clinically effective CRH agonists and antagonists.


Heavy metal resistance patterns of Frankia strains.
The sensitivity of 12 Frankia strains to heavy metals was determined by a growth inhibition assay. In general, all of the strains were sensitive to low concentrations (<0.5 mM) of Ag(1+), AsO(2)(1-), Cd(2+), SbO(2)(1-), and Ni(2+), but most of the strains were less sensitive to Pb(2+) (6 to 8 mM), CrO(4)(2-) (1.0 to 1.75 mM), AsO(4)(3-) (>50 mM), and SeO(2)(2-) (1.5 to 3.5 mM). While most strains were sensitive to 0.1 mM Cu(2+), four strains were resistant to elevated levels of Cu(2+) (2 to 5 mM and concentrations as high as 20 mM). The mechanism of SeO(2)(2-) resistance seems to involve reduction of the selenite oxyanion to insoluble elemental selenium, whereas Pb(2+) resistance and Cu(2+) resistance may involve sequestration or binding mechanisms. Indications of the resistance mechanisms for the other heavy metals were not as clear.


A high-fat diet changes astrocytic metabolism to promote synaptic plasticity and behavior.
A high-fat diet (HFD) is generally considered to negatively influence the body, the brain, and cognition. Nonetheless, fat and fatty acids are essential for nourishing and constructing brain tissue. Astrocytes are central for lipolysis and fatty acids metabolism. We tested how HFD affects astrocyte metabolism, morphology, and physiology. We used Raman microspectroscopy to assess the redox state of mitochondria and lipid content in astrocytes and neurons in hippocampal slices of mice subjected to HFD. Astrocytes were loaded with fluorescent dye through patch pipette for morphological analysis. Whole-cell voltage-clamp recordings were performed to measure transporter and potassium currents. Western blot analysis quantified the expression of astrocyte-specific proteins. Field potential recordings measured the magnitude of long-term potentiation (LTP). Open filed test was performed to evaluate the effect of HFD on animal behavior. We found that exposure of young mice to 1 month of HFD increases lipid content and relative amount of reduced cytochromes in astrocytes but not in neurons. Metabolic changes were paralleled with an enlargement of astrocytic territorial domains due to an increased outgrowth of branches and leaflets. Astrocyte remodeling was associated with an increase in expression of ezrin and with no changes in glial fibrillary acidic protein (GFAP), glutamate transporter-1 (GLT-1), and glutamine synthetase (GS). Such physiological (non-reactive) enlargement of astrocytes in the brain active milieu promoted glutamate clearance and LTP and translated into behavioral changes. Dietary fat intake is not invariably harmful and might exert beneficial effects depending on the biological context.


A structural model of emotions of cognitive dissonances.
Cognitive dissonance is the stress that comes from holding two conflicting thoughts simultaneously in the mind, usually arising when people are asked to choose between two detrimental or two beneficial options. In view of the well-established role of emotions in decision making, here we investigate whether the conventional structural models used to represent the relationships among basic emotions, such as the Circumplex model of affect, can describe the emotions of cognitive dissonance as well. We presented a questionnaire to 34 anonymous participants, where each question described a decision to be made among two conflicting motivations and asked the participants to rate analogically the pleasantness and the intensity of the experienced emotion. We found that the results were compatible with the predictions of the Circumplex model for basic emotions.


Morphology of brachial segments in mudpuppy (Necturus maculosus) spinal cord studied with confocal and electron microscopy.
The isolated brachial spinal cord of Necturus maculosus is useful for studies of neural networks underlying forelimb locomotion, but information about its cellular morphology is scarce. We addressed this issue by using confocal and electron microscopy. Remarkably, the central region of gray matter was aneural and consisted exclusively of a tenuous meshwork of glial fibers and large extracellular spaces. Somata of motoneurons (MNs) and interneurons (INs), labeled by retrograde transport of fluorescent tracers from ventral roots and axons in the ventrolateral funiculus, respectively, were confined within a gray neuropil layer abutting the white matter borders, whereas their dendrites projected widely throughout the white matter. About one-third of labeled INs were found contralaterally, with axons crossing ventral to a thick layer of ependyma surrounding the central canal. Lateral MN dendrites proliferated under the pial surface to form a dense, thin (1-2 microm) plexus immediately beneath a thin layer of glial fibrillary acidic protein-positive glia limitans. The latter contained arrays of unusual tubular structures (diameter 200-400 nm, length 3 microm) that resembled mitochondria but lacked double membranes or cristae. Dorsal roots (DRs) produced dense presynaptic arbors within a wedge-shaped afferent termination zone medial to the dorsal root entry, within which dendrites of MNs and INs mingled with dense collections of synaptic boutons. Our data suggest that a major fraction of synaptic interactions takes place within the white matter. This study provides a detailed foundation for designing electrophysiological experiments to study the neural circuits involved in locomotor pattern generation.


Toca 511 gene transfer and 5-fluorocytosine in combination with temozolomide demonstrates synergistic therapeutic efficacy in a temozolomide-sensitive glioblastoma model.
Toca 511 (vocimagene amiretrorepvec), an amphotropic retroviral replicating vector (RRV), can successfully and safely deliver a functional, optimized cytosine deaminase (CD) gene to tumors in orthotopic glioma models. This agent, in conjunction with subsequent oral extended-release 5-fluorocytosine (5-FC) (Toca FC), is currently under investigation in patients with recurrent high-grade glioma . Temozolomide (TMZ) with radiation is the most frequently used first-line treatment for patients with glioblastoma, the most common and aggressive form of primary brain cancer in adults. However, subsets of patients with certain genetic alterations do not respond well to TMZ treatment and the overall median survival for patients who respond remains modest, suggesting that combinatorial approaches may be necessary to significantly improve outcomes. We show that in vitro TMZ delays but does not prevent RRV spread, nor interfere with Toca 511+5-FC-mediated cell killing in glioma tumor cells, and in vivo there is no significant hematologic effect from the combination of 5-FC and the clinically relevant dose of TMZ. A synergistic long-term survival advantage is observed in mice bearing an orthotopic TMZ-sensitive glioma after Toca 511 administration followed by coadministration of TMZ and 5-FC. These results provide support for the investigation of this novel combination treatment strategy in patients with newly diagnosed malignant glioma.


Oxidised regenerated cellulose: an effective emergency haemostatic in burns surgery.
A 26-year-old man with 40% full-thickness burns was treated by excision and split-skin grafting on the 7th post-burn day. He developed bleeding 1 hour post-operatively which persisted despite the use of pressure dressings, elevation of the limbs, the administration of fresh frozen plasma, platelet-rich plasma, calcium, whole blood and cryoprecipitate respectively over a period of 11 hours. The bleeding was finally controlled under general anaesthesia by clot removal, bipolar electro-coagulation of miniscule oozing points, replacement of the meshed skin graft and the application of a layer of oxidised regenerated cellulose over the grafted area. The properties and uses of oxidised regenerated cellulose are outlined.


Underestimation of glyphosate intake by the methods currently used by regulatory agencies.
The acceptable daily intake (ADI) is an estimate of the amount of a substance in food or beverages that can be consumed daily over a lifetime without presenting an appreciable risk to health. To assess the risk of ingesting glyphosate, regulatory agencies compare glyphosate daily intake to ADI. Based on published data on urine glyphosate levels measured according to known quantities of ingested glyphosate, our objectives were to test the robustness of the mathematical model currently used to calculate glyphosate daily intake, and to propose alternative models based on urinary excretion kinetics. Our results support that the quantity of ingested glyphosate is systematically underestimated by the model currently used by regulatory agencies, whereas the other models evaluated showed better estimations, with differences according to gender. Our results also show a great variability between individuals, leading to some uncertainties notably with regards to the ADI, and further support that glyphosate excretion varies significantly among individuals who follow a similar dosing regimen. In conclusion, our study highlights the lack of reliability of assessment processes carried out by regulatory agencies for glyphosate in particular, and pesticides in general, and questions the relevance of such processes supposed to safeguard human health and the environment.


Intracorneal rings for the correction of pellucid marginal degeneration.
We report a case of Intacs(R) (KeraVision) implantation for the correction of pellucid marginal degeneration (PMD). Preoperatively, the patient's uncorrected visual acuity (UCVA) was 0.05, the best spectacle-corrected visual acuity (BSCVA) was 0.1, and the refraction was -2.00 -7.00 x 90 in the right eye. The flattest meridian (K1) measured 43.8@104 and the steepest meridian (K2), 51.3@14. Ultrasound pachymetry revealed a thin cornea of 420 micrometers. One month postoperatively, the UCVA was 0.2, the BSCVA was 0.5, and the refraction was -8.00 -7.00 x 50. Contact lens best corrected visual acuity was 0.9; 6 months postoperatively, it improved to 1.0. The use of Intacs to treat PMD seems to be viable and improves visual acuity. The residual error can be corrected with contact lenses.


Arthroscopic revisions in failed meniscal surgery.
The aim of this study was to classify meniscal tear forms as found in 195 revision arthroscopies. Interval between primary arthroscopy and revision was 7.8+/-5.6 month. All patients were available for control after 1 year. In 174 knees the lesion was located in the medial meniscus and in 21 knees in the lateral meniscus. In the medial meniscus an unstable posterior meniscal horn was seen in 93 knees followed by incomplete horizontal tear and meniscal destruction in 37. Flap tear, circumferential tear, and failed meniscal repair were also seen. In the lateral meniscus destruction of a discoid meniscus, instability near the popliteal hiatus, and various tear forms were seen with nearly equal frequency. Postoperatively Lysholm score increased significantly in both groups. Most meniscal tears, found in revision arthroscopy, are caused by an insufficient primary operation. A diligent analysis of the tear form is absolutely necessary. An adequate radical resection technique to establish a smooth meniscal crest is indispensable.


Regenerating supernumerary axons are cholinergic and emerge from both autonomic and motor neurons in the rat spinal cord.
Multipolar neurons in the mammalian nervous system normally exhibit one axon and several dendrites. However, in response to an axonal injury, adult motoneurons may regenerate supernumerary axons. Supernumerary axons emerge from the cell body or dendritic trees in addition to the stem motor axon. It is not known whether these regenerating axons contain neurotransmitters for synaptic transmission at their terminals. Here, using immunohistochemistry for choline acetyltransferase, an enzyme that synthesizes acetylcholine, we demonstrate the emergence of cholinergic supernumerary axons at 6 weeks after a unilateral L5-S2 ventral root avulsion and acute implantation of the avulsed L6 ventral root into the adult rat spinal cord. Light microscopic serial reconstruction of choline acetyltransferase immunoreactive arbors shows that these supernumerary axons originate from both autonomic and motor neurons. The supernumerary axons emerge from the cell body or dendrites, exhibit an abnormal projection pattern within the intramedullary gray and white matters, make frequent abrupt turns in direction, and form bouton-like swellings as well as growth cone-like terminals. Double labeling immunohistochemistry studies show that the choline acetyltransferase immunoreactive supernumerary axons co-localized with two proteins associated with axonal growth and elongation, growth-associated protein 43 and p75, the low affinity neurotrophic factor receptor. Our findings suggest that regenerating supernumerary axons selectively transport and store choline acetyltransferase, supporting the notion that supernumerary axons may develop functional and active synaptic transmission. Therefore, regenerating supernumerary axons may contribute to the plasticity in neural circuits following injury in the adult nervous system.


Anin vitro model for the biological evaluation of dental materials using human periodontal ligament cells.
Anin vitro system is described which uses human diploid cells derived from the periodontal ligament. It simulates the clinical situation and is suitable for rapid cytotoxicity screening of dental materials. It can also be adapted to study various factors influencing biocompatibility.


Caldesmon and the structure of smooth muscle thin filaments: immunolocalization of caldesmon on thin filaments.
Antibodies reacting with chicken gizzard caldesmon were used to determine the distribution of caldesmon on smooth muscle thin filaments. Antibodies developed against both the intact caldesmon molecule and a 40 kilodalton proteolytic fragment cause thin filaments to aggregate laterally. Aggregates produced with the latter antibody display regular periodic labelling with a repeat of approximately 38 nm, a distribution characteristic of proteins associated with tropomyosin on thin filaments. The stoichiometry of caldesmon on thin filaments has been critically reevaluated and alternative models of caldesmon distribution on thin filaments are proposed.


Divergent actions of serotonin receptor activation during fictive swimming in frog embryos.
We have investigated the pharmacology underlying locomotor system responses to serotonin (5-HT) in embryos of the frog, Rana temporaria, to provide a comparison to studies in embryos of its close relative, Xenopus laevis. Our findings suggest that two divergent mechanisms underlie the modulation of locomotion by 5-HT in Rana. Bath-applied 5-HT or 5-carboxamidotyptamine, a 5-HT(1,5A,7) receptor agonist, can modulate fictive swimming in a dose-dependent manner, increasing burst durations and cycle periods. However, activation of 5-HT(1,7) receptors with R8-OHDPAT or 8-OHDPAT fails to mimic 5-HT, and in some cases exerts exactly the opposite response; decreasing burst durations and cycle periods. Elevating endogenous 5-HT levels by blocking re-uptake with clomipramine transiently increases burst durations. The receptors involved in this endogenous response include 5-HT(1A) receptors, as in Xenopus, but also 5-HT(7) receptors. However, like the 8-OHDPAT enantiomers, prolonged re-uptake inhibition can result in a motor response in the opposite direction to exogenous 5-HT. This effect is not reversed by 5-HT(1A) and/or 5-HT(7) receptor antagonism, implicating 5-HT(1B/1D) receptors. Remarkably, antagonism of these receptors using methiothepin unmasks a dose-dependent response to clomipramine, reminiscent of exogenous 5-HT. Our data suggest that 5-HT(1A,7) and 5-HT(1B/1D) receptors act as gain-setters of burst durations, whilst 5-HT(5A) receptors are involved in the effects of bath-applied 5-HT on locomotion.


Development of a physiologically based pharmacokinetic model for mefloquine and its application alongside a clinical effectiveness model to select an optimal dose for prevention of malaria in young Caucasian children.
To predict the optimal chemoprophylactic dose of mefloquine in infants of 5-10 kg using physiologically based pharmacokinetic (PBPK) and clinical effectiveness models. The PBPK model was developed in Simcyp version 14.1 and verified against clinical pharmacokinetic data in adults; the final model, accounting for developmental physiology and enzyme ontogeny was then applied in the paediatric population. The clinical effectiveness model utilized real-world chemoprophylaxis data with stratification of output by age and including infant data from the UK population. PBPK simulations in infant populations depend on the assumed fraction of mefloquine metabolized by CYP3A4 (0.47, 0.95) and on the associated CYP3A4 ontogeny (Salem, Upreti). However, all scenarios suggest that a dose of 62.5 mg weekly achieves or exceeds the exposure in adults following a 250 mg weekly dose and results in a minimum plasma concentration of 620 ng ml-1 , which is considered necessary to achieve 95% prophylactic efficacy. The clinical effectiveness model predicts a 96% protective efficacy from mefloquine chemoprophylaxis at 62.5 mg weekly. The PBPK and clinical effectiveness models are mutually supportive and suggest a prophylactic dose of 62.5 mg weekly in the Caucasian 5-10 kg infant population travelling to endemic countries. This dual approach offers a novel route to dose selection in a vulnerable population, where clinical trials would be difficult to conduct.


[Sergentomyia (Sergentomyia) bergerardi n. sp. (Diptera, Phlebotomidae). New species in the Popular Republic of Congo (author's transl)].
Sergentomyia bergerardi, new species collected in the Mayombe forest in the People's Republic of the Congo, is described from two male specimens. This sand fly belongs to the subgenus Sergentomyia and is differenciated from other species of this subgenus from the length of antennal segment III and cibarial armature.


Head and Neck Squamous Cell Carcinoma in Elderly Patients: Role of Radiotherapy and Chemotherapy.
Head and neck squamous cell carcinomas (HNSCC) constitute the sixth most common malignancy worldwide, with approximately 25-40% of the diagnosed patients older than 70 years. HNSCC patients are often frail and frequently have multiple comorbidities due to their unhealthy lifestyle, and evidence suggests that older patients may receive less aggressive and suboptimal treatment than younger patients with the same disease status. The aim of this review is to depict and summarize the evidence regarding the different strategies that can be used in the clinical management of elderly HNSCC patients. Key references were derived from a PubMed query. Hand searching and clinicaltrials.gov were also used. This paper contains a narrative report and a critical discussion of clinical approaches in the context of elderly HNSCC.


Bifurcated endovascular grafting for abdominal aortic aneurysm.
Endovascular grafting represents a minimally invasive approach to prosthetic aortic grafting for abdominal aortic aneurysms (AAA). We reviewed our initial experience with a bifurcated endovascular graft that shares with conventional grafts the characteristics of discrete transaortic fixation and unitary unsupported woven polyester construction. Twenty-eight patients (26 male, 2 female; age: 58-93) with infrarenal aortic aneurysms between 41 and 82 mm in greatest diameter (x = 55.4 mm) underwent bifurcated endovascular grafting (Guidant/EVT, Menlo Park, CA) over an 18-month period. We concluded that bifurcated endovascular grafting with the EVT(R) device provides reliable and reproducible aneurysm exclusion with short hospital recovery and low morbidity.


Ozz; a new name on the long list of beta-catenin's nemeses.
In the February issue of Developmental Cell, Nastasi et al. describe Ozz, a muscle-specific ubiquitin ligase adaptor that regulates myofibril organization. Ozz appears to function in ubiquitination and degradation of membrane-bound, but not cytosolic, beta-catenin, whose turnover may be required for alignment and growth of the sarcomere, the basic contractile unit of myofibers.


Dynamic Target Definition: a novel approach for PTV definition in ion beam therapy.
To present a beam arrangement specific approach for PTV definition in ion beam therapy. By means of a Monte Carlo error propagation analysis a criteria is formulated to assess whether a voxel is safely treated. Based on this a non-isotropical expansion rule is proposed aiming to minimize the impact of uncertainties on the dose delivered. The method is exemplified in two cases: a Head and Neck case and a Prostate case. In both cases the modality used is proton beam irradiation and the sources of uncertainties taken into account are positioning (set up) errors and range uncertainties. It is shown how different beam arrangements have an impact on plan robustness which leads to different target expansions necessary to assure a predefined level of plan robustness. The relevance of appropriate beam angle arrangements as a way to minimize uncertainties is demonstrated. A novel method for PTV definition in on beam therapy is presented. The method show promising results by improving the probability of correct dose CTV coverage while reducing the size of the PTV volume. In a clinical scenario this translates into an enhanced tumor control probability while reducing the volume of healthy tissue being irradiated.


Large-scale lysine crotonylation analysis reveals the role of TRAF6-Ecsit complex in endoplasmic reticulum stress in mud crab (Scylla paramamosain).
Lysine crotonylation (Kcr) is a newly discovered type of post-translational modification. Although Kcr has been reported in several species, its role in crustaceans remains largely unknown. In this study, Kcr in hemocytes of mud crab (Scylla paramamosain) was characterized using pan anti-crotonyllysine antibody enrichment and high-resolution liquid chromatogram-mass spectrometry analysis after SpTRAF6 or SpEcsit silencing. Altogether, 1,800 Kcr sites with six conserved motifs were identified from 512 proteins. Subcellular localization analysis showed that the identified Kcr proteins were mainly localized to the cytoplasm, nucleus, and mitochondria. The cellular components analysis showed that the 'chromosomal region' was enriched in the hemocytes of SpTRAF6-or SpEcsit-silenced mud crabs. The KEGG and PPI analyses showed that the identified Kcr proteins in the hemocytes SpTRAF6-or SpEcsit-silenced mud crabs were related to the 'protein processing in endoplasmic reticulum'; of which the marker of endoplasmic reticulum stress (Bip) was identified to be crotonylated. These datasets present the first comprehensive analysis of the crotonylome in mud crab hemocytes, providing invaluable insights into the regulatory functions of SpTRAF6 and SpEcsit in Kcr. Additionally, our findings shed light on the potential role of these proteins in activating marker proteins during endoplasmic reticulum stress in invertebrates.


Variable presentation of the clinical phenotype of McArdle's disease in a kindred harbouring a novel compound genotype in the muscle glycogen phosphorylase gene.
We report a Spanish family with muscle glycogen phosphorylase (PYGM) deficiency (McArdle's disease) harbouring a novel compound genotype (A659D/L586P). Four individuals who had the same genotype for PYGM, showed a wide variability in the presentation of the clinical phenotype, including one patient with a restrictive respiratory pattern, which is unusual in McArdle's disease. Moreover, these patients were studied for the insertion/deletion (I/D) trait in the angiotensin converting enzyme (ACE) which has been suggested to be a strong modulator of severity in McArdle's disease. Our results indicate no association of the I/D ACE trait in this family, suggesting that other factors would be more relevant in determining the severity of the clinical presentation.


Prevalence and factors associated with severe undernutrition among under-5 children in Bangladesh, Pakistan, and Nepal: a comparative study using multilevel analysis.
Despite economic growth and poverty reduction, under-5 child undernutrition is still rampant in South Asian countries. This study explored the prevalence and risk factors of severe undernutrition among under-5 children in Bangladesh, Pakistan, and Nepal for comparison using the Composite Index of Severe Anthropometric Failure. We utilised information on under-5 children from recent Demographic Health Surveys. We used multilevel logistic regression models for data analysis. The prevalence of severe undernutrition among under-5 children was around 11.5%, 19.8%, and 12.6% in Bangladesh, Pakistan, and Nepal, respectively. Children from the lowest socioeconomic quintile, and children born with low birth weight were key factors associated with severe undernutrition in these countries. The factors, parental education, maternal nutritional status, antenatal and postnatal care, and birth order were not homogeneous in explaining the determinants of child severe undernutrition across the countries. Our results suggest that the poorest households, and low birth weight of children have significant effects on severe undernutrition among under-5 children in these countries, which should be considered to formulate an evidence-based strategy to reduce severe undernutrition in South Asia.


Rapid prenatal diagnosis of trisomies 13, 18, 21 and sex chromosome anomalies by fluorescence in situ hybridization: a year's experience.
The present paper summarizes our first 12 months' experiences of rapid prenatal diagnosis using commercially available diagnostic fluorescence in situ hybridization (FISH) probes for chromosomes X, Y, 13, 18 and 21. The data clearly demonstrate that the advantage of using FISH as an adjunct technique is the fast and reliable determination of the common fetal chromosomal aneuploidies; the results are available in less than 24 hours instead of the 7-14 days with standard techniques.


Changing within-trial array location and target object position enhances rats' (Rattus norvegicus) missing object recognition accuracy.
Six rats were trained to find a previously missing target or 'jackpot' object in a square array of four identical or different objects (the test segment of a trial) after first visiting and collecting sunflower seeds from under the other three objects (the study segment of a trial). During training, objects' local positions within the array and their global positions within the larger foraging array were varied over trials but were not changed between segments within a trial. Following this training, rats were tested on their accuracy for finding the target object when a trial's test array was sometimes moved to a different location in the foraging arena or when the position of the target object within the test array had been changed. Either of these manipulations initially slightly reduced rats' accuracy for finding the missing object but then enhanced it. Relocating test arrays of identical objects enhanced rats' performance only after 10-min inter-segment intervals (ISIs). Relocating test arrays of different objects enhanced rats' performance only after 2-min ISIs. Rats also improved their performance when they encountered the target object in a new position in test arrays of different objects. This enhancement effect occurred after either 2- or 30-min ISIs. These findings suggest that rats separately retrieved a missing (target) object's spatial and non-spatial information when they were relevant but not when they were irrelevant in a trial. The enhancement effects provide evidence for rats' limited retrieval capacity in their visuo-spatial working memory.


Increasing HIV early diagnosis by implementing an automated screening strategy in emergency departments.
Late HIV diagnosis is associated with increased morbidity, mortality and risk of onward transmission. Increasing HIV early diagnosis is still a priority. In this observational study with historical control, we determined the impact of an opportunistic HIV screening strategy in the reduction of late diagnosis and missed opportunities for earlier diagnosis. The screening programme was implemented in the emergency department (ED) of the Hospital de Cascais between September 2018 and September 2021. Eligible patients were aged 18-64 years, with no known HIV diagnosis or antibody testing performed in the previous year, and who required blood work for any reason. Out of the 252 153 emergency visits to the ED, we identified 43 153 (17.1%) patients eligible for HIV testing. Among the total population eligible for the screening, 38 357 (88.9%) patients were ultimately tested for HIV. Impact of the ED screening was determined by analysing late diagnosis in the ED and missed opportunities at different healthcare settings 3 years before and 3 years after the start of the ED screening. After 3 years of automated HIV ED testing, we found 69 newly diagnosed HIV cases (54% male, 39% Portuguese nationals, mean age 40.5 years). When comparing the characteristics of HIV diagnoses made in the ED, we observed a significant reduction in the number of people with late HIV diagnosis before and after implementation of the screening programme (78.4% vs. 39.1%, respectively; p = 0.0291). The mean number of missed opportunities for diagnosis also fell (2.6 vs. 1.5 annual encounters with the healthcare system per patient, p = 0.0997). People living with HIV in Cascais and their providers miss several opportunities for earlier diagnosis. Opportunistic screening strategies in settings previously deemed to be unconventional, such as EDs, are feasible and effective in mitigating missed opportunities for timely HIV diagnosis.


Chronic and recurrent choroidal detachment after glaucoma filtering surgery.
Chronic and recurrent choroidal (ciliochoroidal) detachments developed following glaucoma filtration surgery in 14 eyes of 13 patients during a 9-year period. Three specific subgroups were identified: recurrent, inflammatory, and chronic (present for more than 6 months). The factors that may be related to the development of chronic and recurrent choroidal detachments included patient age (mean, 68.8 years), systemic hypertension or atherosclerotic heart disease, hyperopia, aqueous suppressant therapy, ocular inflammation, and full-thickness filtration surgery. A total of 46 choroidal detachments in 14 eyes were recorded and required drainage of suprachoroidal fluid on 34 occasions. All eyes developed visually significant cataracts, and complete resolution of the recurrent or chronic choroidal detachment occurred following cataract extraction in six eyes. Treatment of chronic and recurrent choroidal detachments should include intense therapy of ocular inflammation, discontinuation of medications that can incite ocular inflammation, discontinuation of topical and systemic aqueous suppressant therapy, and when a visually significant cataract is present, cataract extraction combined with a choroidal tap should be performed.


Comparison of adjuvant emulsions for their safety and ability to enhance the antibody response in horses immunized with African snake venoms.
Adjuvant emulsions are widely used to enhance the antibody response in animals used as immunoglobulin source to produce snake antivenoms. We tested the performance of four commercial emulsion adjuvants (Montanide, Freund, Carbigen, and Emulsigen-D) and an experimental adjuvant (QH-769) in the antibody response of horses towards venoms of the African snakes Bitis arietans, Echis ocellatus, Dendroaspis polylepis and Naja nigricollis. Montanide, Freund and Carbigen adjuvants generated the highest immune response but induced moderate/severe local lesions at the site of injection. In contrast, Emulsigen-D and QH-769 adjuvants generated the lowest immune response and low incidence of local lesions. No evidence of systemic alterations was observed in the horses immunized with any of the adjuvants. It is suggested that the use of Montanide or Freund-based emulsions in the first immunization steps, followed by the use of Emulsigen-D, QH-769 or similar adjuvants in the following injections, could result in a satisfactory immune response against snake venoms, while not inducing serious local deleterious effects.


Classification images of two right hemisphere patients: a window into the attentional mechanisms of spatial neglect.
While spatial neglect most commonly occurs after right hemisphere lesions, damage to diverse areas within the right hemisphere may lead to neglect, possibly through different mechanisms. To identify potentially different causes of neglect, the visual information used (the 'perceptual template') in a cueing task was estimated with a novel technique known as 'classification images' for five normal observers and two male patients with right-hemisphere lesions and previous histories of spatial neglect (CM, age 85; HL, age 69). Observers made a yes/no decision on the presence of a 'White X' checkerboard signal (1.5 degrees ) at one of two locations, with trial-to-trial stimulus noise added to the 9 checkerboard squares. Prior to the stimulus, a peripheral precue (140 ms) indicated the signal location with 80% validity. The cueing effects and estimated perceptual templates for the normal observers showed no visual field differences. Consistent with previous studies of spatial neglect, both patients had difficulty with left (contralesional) signals when preceded by a right (ipsilesional) cue. Despite similar behavioral results, the patients' estimated perceptual templates in the left field suggested two different types of attentional deficits. For CM, the left template matched the signal with left-sided cues but was opposite in sign to the signal with right-sided cues, suggesting a severely disrupted selective attentional strategy. For HL, the left templates indicated a general uncertainty in localizing the signal regardless of the cue's field. In conclusion, the classification images suggested different underlying mechanisms of neglect for these two patients with similar behavioral results and hold promise in further elucidating the underlying attentional mechanisms of spatial neglect.


Prevalence and outcomes of thrombotic and hemorrhagic complications in pediatric acute promyelocytic leukemia in a tertiary Brazilian center.
Little attention is given to thrombosis associated with pediatric acute promyelocytic leukemia (APL). This study describes the thrombotic and hemorrhagic manifestations of APL in pediatric patients and evaluates their hemostasis, based on coagulation tests. Inclusion criteria were age 0-18 years and APL diagnosis between April 2005 and November 2017. Patients who had received blood transfusion prior to coagulation tests were excluded. Baseline coagulation tests, hematologic counts, and hemorrhagic/thrombotic manifestations were evaluated. Median age was 10.7 years (1-15 years). The initial coagulation tests revealed a median Hgb of 8.3 g/dL (4.7-12.9 g/dL), median leucocyte count of 10.9 × 10⁹/L (1.1-95.8 × 10⁹/L), median platelet count of 31.8 × 10⁹/L (2.0-109.0 × 10⁹/L), median activated partial thromboplastin time (aPTT) of 31.7 s (23.0-50.4 s), median aPTT ratio of 1.0 (0.78-1.6), median thromboplastin time (PT) of 17.5 s (13.8-27.7 s), median PT activity of 62% (25-95 %), and median fibrinogen of 157.7 mg/dL (60.0-281.0 mg/dL). Three patients (13%) had thrombosis. At diagnosis, 21 patients (91.3%) had bruising, one patient (4.3%) had splenic vein and artery thrombosis and one patient (4.3%) presented without thrombohemorrhagic manifestations. During treatment, two patients (8.6%) had thrombosis. Knowledge of thrombosis in pediatric APL is important to determine its risk factors and the best way to treat and prevent this complication.


Stress in conic piles determined by a centrifuge experiment: breakdown of scaling hypothesis.
It is found experimentally that vertical-stress field in a conic pile depends on gravity level, building process, and loading story. For instance, a conic pile with inclined strata does exhibit a minimum of stress in the center, whereas conic pile with horizontal strata does not; both piles exhibit an arching effect, which increases with gravity. This questions the assumptions of radius stress field scaling. Amplitude of the stress dip is found to be 10%, which is much smaller than what was found in previous experiments.


A case study in participatory science with mutual capacity building between university and tribal researchers to investigate drinking water quality in rural Maine.
Participatory science or citizen science is increasingly being recognized for providing benefits to scientists and community members alike. However, most participatory science projects include community researchers only in the sample collection phase of the research project. Here we describe how a rural tribal community and urban university utilized participatory science methods to engage community researchers across an entire research study, creating numerous opportunities for mutual capacity building. Researchers from MIT and the Sipayik Environmental Department, a tribal government department, partnered to co-launch a participatory science project to analyze municipal and private well drinking water quality in households in three Maine communities. The objective was to provide households with information about metals, primarily lead and arsenic, in their drinking water, and to improve public education, community partnerships, and local scientific capacity. MIT and Sipayik researchers engaged local communities through public community meetings, mailed flyers sent to residents, and meetings with local stakeholders. MIT and community researchers worked together to design and implement the study to quantify metals in community drinking water samples, as well as hold capacity-building trainings. Individual drinking water results were communicated to households, and generalized results were discussed at community meetings in the report-back phase. The study attained a 29% household participation rate in the region. The researchers completed the analysis and report-back on 652 water samples. Isolated incidences of lead and geologically-attributable arsenic exceeding EPA standards were found. Individual report-backs of the results enabled local participatory scientists to make their own informed public health decisions. The study produced methodologies for navigating potential ethical issues, working with diverse communities, and collaborating over challenging geographical distances. This project developed methodologies to build long-term relationships with local scientists and to engage community members and enhance the environmental literacy of rural communities. Both MIT and Sipayik researchers learned from each other throughout the project; Sipayik researchers built technical capacity while MIT researchers gained local and cultural understanding. Community outreach methods were most effective when sent directly to residents as mailed flyers or through Sipayik researchers' outreach.


Characterization of bacterial communities in Coregonus peled fillets during chilled storage and interactions between selected bacterial strains.
Coregonus peled fillets were used as a model to evaluate the dominant bacterial growth of chilled fish during storage after shipping and interactions of selected bacterial strains. Coregonus peled fillets were transported by air and land in ice boxes about 48 h from aquatic products company in Xinjiang, China, to the laboratory located in Dalian, China. Both culture-dependent (plate counts on nonselective media) based on 16S rRNA gene sequencing and culture-independent (Illumina-MiSeq high-throughput sequencing) methods were used. To detect interactions among bacterial populations from chilled fish, the influence of 18 test strains on the growth of 12 indicator isolates was measured by a drop assay and in liquid culture medium broth. The results showed that bacterial counts exceeded 7.0 log CFU/g following storage for 4 days at 4 °C. When the bacterial counts exceeded 8.5 log CFU/g after 12 days, the predominant micro-organisms were Aeromonas, Pseudomonas, Carnobacterium, Psychrobacter and Shewanella, as measured by the culture-independent method. All test strains showed inhibiting effects on the growth of other strains in liquid culture. Pseudomonas isolates showed antibacterial activity for approximately 60% of the indicator strains on nutritional agar plates. The majority of test isolates enhancing indicator strain growth were the strains isolated on day 0. High-throughput sequencing approach gives whole picture of bacterial communities in chilled C. peled fillets during storage, while growth interferences between selected bacterial strains illustrate the complexity of microbial interactions. We determined the bacterial communities and growth interferences in chilled Coregonus peled after shipping and these are the first data concerning microbiota in C. peled using a culture-independent analysis. The present study will be useful for manufacture and preservation of C. peled products by providing with valuable information regarding microbiological spoilage of C. peled.


Corticofugal Modulation of DPOAEs in Gerbils.
Efferent auditory feedback on cochlear hair cells is well studied regarding olivocochlear brainstem mechanisms. Less is known about how the descending corticofugal system may shape efferent feedback and modulate cochlear mechanics. Distortion-product otoacoustic emissions (DPOAEs) are a suitable tool to assess outer hair cell function, as they are by-products of the nonlinear cochlear amplification process. The present project investigates the effects of cortical activity on cubic and quadratic DPOAEs in mongolian gerbils, Meriones unguiculatus, through cortical deactivation using the sodium-channel blocker lidocaine. Contralateral cortical microinjections of lidocaine can lead to either an increase or decrease of median DPOAE levels of up to 10.95 dB. The effects are reversible and comparable at all tested frequencies (0.5-40 kHz). They are not restricted to the preferred frequency of the cortical site of injection. Recovery times are between 20 and 120 min depending on stimulation levels and emission type. When the injection is performed in the ipsilateral hemisphere, DPOAE level shifts are lower in amplitude compared to those after injection in the contralateral hemisphere. No significant changes in DPOAE levels are obtained after saline microinjections. Results indicate that deactivation of auditory cortex activity through lidocaine has a considerable impact on peripheral auditory responses in form of DPOAEs, probably through cortico-olivocochlear pathways.


ANP32A regulates ATM expression and prevents oxidative stress in cartilage, brain, and bone.
Osteoarthritis is the most common joint disorder with increasing global prevalence due to aging of the population. Current therapy is limited to symptom relief, yet there is no cure. Its multifactorial etiology includes oxidative stress and overproduction of reactive oxygen species, but the regulation of these processes in the joint is insufficiently understood. We report that ANP32A protects the cartilage against oxidative stress, preventing osteoarthritis development and disease progression. ANP32A is down-regulated in human and mouse osteoarthritic cartilage. Microarray profiling revealed that ANP32A protects the joint by promoting the expression of ATM, a key regulator of the cellular oxidative defense. Antioxidant treatment reduced the severity of osteoarthritis, osteopenia, and cerebellar ataxia features in Anp32a-deficient mice, revealing that the ANP32A/ATM axis discovered in cartilage is also present in brain and bone. Our findings indicate that modulating ANP32A signaling could help manage oxidative stress in cartilage, brain, and bone with therapeutic implications for osteoarthritis, neurological disease, and osteoporosis.


Purification and characterization of the 47,000-dalton protein phosphorylated during degranulation of human platelets.
Secretion of platelet granule constituents is closely associated with the phosphorylation of a cytosol polypeptide of Mr = 47,000 that we have called P47 (Haslam, R. J., Lynham, J. A., and Fox, J. E. B. (1979) Biochem. J. 178, 397-406). This polypeptide is a substrate of Ca2+-activated phospholipid-dependent protein kinase (Kawahara, Y., Takai, Y., Minakuchi, R., Sano, K., and Nishizuka, Y. (1980) Biochem. Biophys. Res. Commun. 97, 309-317). Two-dimensional gel electrophoresis of protein from human platelets that had been preincubated with 32Pi demonstrated the presence under control conditions of 2-3 major forms of P47 that contained very little 32P (pI values, 6.6-6.8) and, after induction of secretion with thrombin, their replacement by 7-9 highly labeled phosphorylated forms of P47 (pI values, 6.1-6.5). Native phosphorylated P47 was purified from thrombin-stimulated 32P-labeled platelets by ammonium sulfate fractionation and column chromatography on DEAE-cellulose, phenyl-Sepharose, and hydroxylapatite. The final 32P-labeled product was obtained in a yield of 20-25% and was purified about 400-fold relative to platelet lysate. This material was homogeneous on sodium dodecyl sulfate-polyacrylamide gel electrophoresis but, like the starting material, contained 7-9 separable phosphorylated components with different pI values. Purified phosphorylated P47 had a sedimentation coefficient (s20,w) of 3.57 S and a Stokes radius of 3.33 nm from which an Mr = 49,000 and a frictional ratio (f/f0) of 1.4 were calculated. These findings and failure to detect multimers after treatment of the protein with dimethyl suberimidate indicate that P47 normally exists as a monomer. The 32P-labeled phosphate present in purified P47 had the chemical stability of serine or threonine phosphoesters and analysis indicated the presence of 83% phosphoserine and 17% phosphothreonine. Limited proteolysis of purified 32P-labeled P47 by Staphylococcus aureus V8 protease generated a major unlabeled fragment (Mr = 23,500) and up to six labeled fragments (Mr = 24,700-14,800), the relative amounts of the latter depending on the extent of proteolysis. The same labeled fragments were obtained after proteolysis of each of the major phosphorylated components of P47, suggesting that these represent different phosphorylation states of variants of the same protein and that most or all of the phosphorylation sites are on a single 14,800-Da segment of the protein. The availability of pure native phosphorylated P47 should facilitate investigation of the physiological role of this protein in platelets.


Suggestion does not de-automatize word reading: evidence from the semantically based Stroop task.
Recent studies have shown that the suggestion for participants to construe words as meaningless symbols reduces, or even eliminates, standard Stroop interference in highly suggestible individuals (Raz, Fan, & Posner, 2005; Raz, Kirsch, Pollard, & Nitkin-Kaner, 2006). In these studies, the researchers consequently concluded that this suggestion de-automatizes word reading. The aim of the present study was to closely examine this claim. To this end, highly suggestible individuals completed both standard and semantically based Stroop tasks, either with or without a suggestion to construe the words as meaningless symbols (manipulated in both a between-participants [Exp. 1] and a within-participants [Exp. 2] design). By showing that suggestion substantially reduced standard Stroop interference, these two experiments replicated Raz et al.'s (2006) results. However, in both experiments we also found significant semantically based Stroop effects of similar magnitudes in all suggestion conditions. Taken together, these results indicate that the suggestion to construe words as meaningless symbols does not eliminate, or even reduce, semantic activation (assessed by the semantically based Stroop effect) in highly suggestible individuals, and that such an intervention most likely reduces nonsemantic task-relevant response competition related to the standard Stroop task. In sum, contrary to Raz et al.'s claim, suggestion does not de-automatize or prevent reading (as shown by a significant amount of semantic processing), but rather seems to influence response competition. These results also add to the growing body of evidence showing that semantic activation in the Stroop task is indeed automatic.


Trace elements in the silky shark Carcharhinus falciformis in the Central Pacific Mexican Shelf.
Mercury (Hg), cadmium (Cd) and copper (Cu) concentrations were determined in muscle, liver and fin of the silky shark (Carcharhinus falciformis) caught in the Central Pacific Mexican Shelf. Liver tissue presented the highest concentrations of Cd (2.83 μg g-1 wet weight (ww)) and Cu (2.34 μg g-1 ww). For muscle and fin, Cu presented the highest concentrations (0.97 and 1.80 μg g-1 ww, respectively). Liver concentrations were influenced by the maturation stages for the three elements. Immature organisms exhibited lower trace element concentrations than adults. No significant differences were found between sexes, except for Cu concentrations in muscle, where adult females present higher levels than adult males. One muscle sample (2.3 %) exceeded the Mexican Legislation for Hg (1.0 μg g-1 ww) and five muscles samples (11.6 %) exceeded the Mexican limit for Cd (0.5 μg g-1 ww). Meat consumption of this species can be a risk to human health if it is ingested regularly.


An improved formula for the judgement of intrathecally produced IgG in the presence of blood brain barrier damage.
In order to develop a better formula for discrimination between transsudated and intrathecally produced IgG, we studied the relation between cerebrospinal fluid (CSF) IgG, serum IgG, CSF albumin, and serum albumin for patients with blood brain barrier (BBB) damage. From this study we derived a new formula, IgG extended index = (CSF IgG/serum IgG)/(CSF albumin/serum albumin). Applying this index on 152 patients with BBB damage, but not expected intrathecal IgG synthesis, yielded 14 patients (9.2%) with increased (greater than 1.24) IgG extended index. In comparison, 'conventional' IgG index was increased in 34 of the patients (22%). The diagnostic sensitivity for 68 patients with multiple sclerosis (MS), did not differ significantly between the two formulas (81 and 87%, respectively). A formula for the estimation of 24 hours IgG production was also developed, but this formula was not better than IgG index. Children (0.5-16 yr) had similar IgG patterns as adults.


Induction of growth cessation by acacetin via β-catenin pathway and apoptosis by apoptosis inducing factor activation in colorectal carcinoma cells.
Acacetin, a bioflavanoid, contains anti-inflammatory and anti-cancer activities as shown in different experimental models. However, its anticancer potential and mechanism of action against colorectal cancer cells is largely unknown. Here, we have investigated the efficacy of acacetin using two colorectal adenocarcinoma SW480 and HCT-116 cell lines. Cell survival was examined by Trypan-blue exclusion and MTT assays, cell cycle analysis by FACS, apoptosis was assessed using Annexin V FITC assay and nuclear condensation by Hoechst staining, ROS level by DCFDA and Mitosox, and protein expression level by Western blotting. Acacetin reduced the cell survival and proliferation of both types of cells, and induced S- and G2-M phase arrest and also reduced the levels of β-catenin and its downstream target c-myc. Further, acacetin induced apoptosis as examined by Annexin-V FITC and nuclear condensation. It increased intracellular ROS production, especially mitochondrial ROS. Acacetin increased mitochondrial membrane potential depolarization and Bax:Bcl-2 ratio. Although significant changes in caspases -8 and -9 and PARP level was not observed, acacetin could induce the truncation and subsequent translocation of activated AIF from mitochondria to cytosol, which could further induce chromosomal breakage leading to apoptosis. In conclusion, Acacetin induces mitochondrial ROS-mediated cell death in a caspase-independent manner in SW480 and HCT-116 colon carcinoma cells by inducing apoptosis inducing factor (AIF), which may potentiate its anticancer and chemotherapeutic prospects against colorectal carcinoma.


TERMINAL FLOWER1 is a breeding target for a novel everbearing trait and tailored flowering responses in cultivated strawberry (Fragaria × ananassa Duch.).
The effects of daylength and temperature on flowering of the cultivated octoploid strawberry (Fragaria × ananassa Duch.) have been studied extensively at the physiological level, but information on the molecular pathways controlling flowering in the species is scarce. The flowering pathway has been studied at the molecular level in the diploid short-day woodland strawberry (F. vesca L.), in which the FLOWERING LOCUS T1 (FvFT1)-SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (FvSOC1)-TERMINAL FLOWER1 (FvTFL1) pathway is essential for the correct timing of flowering. In this work, we show by transgenic approach that the silencing of the floral repressor FaTFL1 in the octoploid short-day cultivar 'Elsanta' is sufficient to induce perpetual flowering under long days without direct changes in vegetative reproduction. We also demonstrate that although the genes FaFT1 and FaSOC1 show similar expression patterns in different cultivars, the regulation of FaTFL1 varies widely from cultivar to cultivar and is correlated with floral induction, indicating that the transcription of FaTFL1 occurs at least partially independently of the FaFT1-FaSOC1 module. Our results indicate that changing the expression patterns of FaTFL1 through biotechnological or conventional breeding approaches could result in strawberries with specific flowering and runnering characteristics including new types of everbearing cultivars.


[A simple method of extracorporeal membrane oxygenation (ECMO)--3. The effects of pumpless ECMO (arterio-venous shunt) on arterial blood gas measured for 24 hours].
In eight anesthetized mongrel dogs, the pumpless ECMO applied between the femoral artery and vein was performed under the condition of hypoventilation for 24 hours. The methods were same as the first and second reports, except the pumpless ECMO was used. The abnormal parameters in the cardiovascular and respiratory system (blood gas analysis and end expiratory gas analysis), induced by hypoventilation, recovered to almost normal ranges, by operation of the pumpless ECMO for 24 hours. The pumpless ECMO might have such advantages over ECMO with pump as the less destruction of blood cell, easy performance and simple apparatus. In conclusion, pumpless ECMO using arterio-venous shunt may be applied clinically on respiratory distress conditions.


[Analysis of dengue fever deaths in Mexico: 2009].
to describe the dengue fever mortality. a descriptive and retrospective study including 104 files reported deaths caused by dengue fever during 2009 to march 2010, was done. sixty (58 %) were women and 44 (42 %) men. An increased mortality between the ages of 11 and 40 years old (47 %) was observed. Colima was a state with high incidence of cases and Jalisco had the highest mortality. Thrombocytopenia was the rule (90.4 %) and in one third of the cases platelets were below 50,000/mm(3). A quarter of cases were associated with comorbility. The initial clinical manifestations included: bleeding, hypovolemia by depletion or hemorrhage, tachycardia, paleness, depressed level of consciousness and circulatory failure. The main cause of death was hypovolemic shock or sepsis. In 42 cases, severe dengue was considered. an association between the severity of dengue fever and mortality was observed. The main cause of mortality was a shock state.