Photoperiod sensitivity of the Arabidopsis circadian clock is tissue-specific.
Tissue-specific functions of the circadian clock in Arabidopsis have recently been revealed. The vasculature clock shows distinctive gene expression profiles compared to the clock in other tissues under light-dark cycles. However, it has not yet been established whether the vasculature clock also shows unique gene expression patterns that correlate with temperature cycles, another important environmental cue. Here, we detected diel phase of TIMING OF CAB EXPRESSION 1 (TOC1) expression in the vasculature and whole leaf under long-day light-dark cycles and temperature cycles. We found that the vasculature clock had advanced TOC1 phase under light-dark cycles but not under temperature cycles, suggesting that the vasculature clock has lower sensitivity against temperature signals. Furthermore, the phase advancement of TOC1 was seen only under long-day condition but not under short-day condition. These results support our previous conclusion that the circadian clock in vasculature preferentially senses photoperiodic signals.


Mandatory Medication Indications in Electronic Systems - The Prescriber Perspective.
As hospitals transition from paper to electronic medication charts, an opportunity exists to 'nudge' prescribers to document medication indications by making this data-entry field mandatory. The aim of this study was to explore hospital doctors' perceptions of mandatory documentation of indications in an electronic medication management (EMM) system. Ten junior doctors took part in brief semi-structured interviews. Participants identified improved communication among staff as a key benefit of indication documentation. Recording indications was also seen to act as a prompt for medication review. Despite these benefits, indication documentation for all medications would be challenging to implement in practice. Users of the EMM system (i.e. junior doctors) explained that they are time poor and are often tasked with transcribing medication orders into the electronic system with limited knowledge of why medications are being prescribed. Determining the indication for use would require additional time and effort, and prescribers reported a high risk of working around the system if indication documentation was made mandatory.


Differential response to infusions of highly purified and conventional bovine and porcine insulins.
The glucose clamp technique allows quantification of tissue sensitivity to insulin without modulation by the effects of hypoglycemia. The amount of glucose infused to maintain euglycemia during an insulin infusion gives a direct measure of tissue sensitivity. We have used this technique to compare the metabolic effects of highly purified porcine, highly purified bovine, and conventional insulins in diabetic subjects: (A) a group normally taking conventional bovine insulin, and (B) diabetics who had originally been treated with conventional bovine insulin and had changed to highly purified porcine insulin. Subjects were studied while attached to Biostator with blood glucose values clamped at the basal level during a 2-h insulin infusion (0.01 U/kg/h). both groups of diabetics received significantly more glucose in the first hour of the monocomponent porcine insulin infusion than during the same period either of conventional or highly purified bovine insulin infusion. Highly purified porcine insulin infusion was also associated with a more rapid fall in blood ketone body concentrations. Highly purified porcine insulin infusion was accompanied by a greater and earlier increase in free insulin concentrations. All diabetics had antibodies to insulin but no differential antibody binding of beef and pork insulin was found in the two groups of patients. Thus, use of highly purified porcine insulin is associated with a more rapid onset of action and a greater initial increment in free insulin values.


Management of the uncomplicated postdate pregnancy.
The management of a postdate (greater than 42 weeks) pregnancy with no apparent complication is often unclear. This study compares the intrapartum courses and perinatal outcomes between 101 postdate pregnancies and 322 term pregnancies. The mean maternal age, race, parity and type of care service were not significantly different between the two groups. Signs of fetal compromise were also not different, but a large fetus was more common (p less than 0.01) in the postdate pregnancies (16 cases). The need for cesarean section (13 cases) or midforceps manipulation (7 cases) was increased fourfold in the postdate over the term pregnancies (p less than 0.05). Meconium during labor was not more common in the postdate pregnancies, but the one postdate perinatal death occurred in the neonatal period from severe meconium aspiration complications. A postdate pregnancy with no apparent complications may safely be observed closely.


Sporamin suppresses growth of human esophageal squamous cell carcinoma cells by inhibition of NF‑κB via an AKT‑independent pathway.
The aim of the present study was to determine whether sporamin, a trypsin inhibitor, suppresses the growth of human esophageal squamous cell carcinoma (ESCC) cells in vitro. Sporamin treatment led to the suppression of viability and proliferation of human ESCC cell lines, EC9706 and EC109, as determined by MTT and [3H] thymidine incorporation assays, respectively. Flow cytometry and fluorescence microscopy demonstrated that sporamin significantly induced apoptosis in EC9706 and EC109 cells. Western blotting demonstrated that sporamin downregulated the expression of Bcl‑2 and Bcl‑2 like 1, and upregulated the expression of Bcl‑2‑associated X in EC9706 and EC109 cells. In addition, marked inhibition of nuclear factor (NF)‑κB activation was observed in sporamin‑treated EC9706 and EC109 cells by an electrophoretic mobility shift assay. Sporamin treatment also resulted in reduced expression levels of phosphorylated (p)‑NF‑κB inhibitor α and nuclear NF‑κB p65. However, the expression levels of p‑protein inase (AKT) and its downstream target, p‑p70 S6 kinase, were not markedly altered following sporamin treatment. In conclusion, sporamin may suppress the growth of human ESCC cells via NF‑κB‑dependent and AKT‑independent mechanisms and may act as a promising natural therapeutic agent for the treatment of human ESCC.


Differences in Trabecular Bone, Cortical Shell, and Endplate Microstructure Across the Lumbar Spine.
The quality of the vertebral body structures such as endplate, cortex, and trabecular bone is important for understanding the performance of implants, particularly at the bone-implant interface. Although vertebral body structures have been analyzed separately in the literature, there is no comprehensive study to assess these anatomical measurements along with their interrelationships in the lumbar spine. Therefore, the purpose of this study was to assess variations in trabecular microstructure, vertebral endplate thickness and concavity, and vertebral body cortex thickness within the lumbar spine. A total of 80 lumbar vertebrae (L1-L5) were dissected from 16 human cadaver specimens and imaged with microcomputed tomography to determine trabecular microstructure, vertebral cortex thickness, endplate thickness, and maximum endplate concavity depth. A paired t test and regression analysis were used to determine significant differences (P < .05) between different vertebral levels and correlations between the analyzed anatomical parameters. L1 vertebra had significantly better (P < .02) trabecular bone microstructure (eg, trabecular bone volume fraction) than all other lumbar vertebrae. However, L1 vertebra also had significantly thinner (P ≤ .02) anterior, left, and right cortices compared to all other vertebral levels. Within L3-L5 intervertebral disc spaces, cranial endplates had significantly greater (P ≤ .03) thickness and maximum concavity depth compared to their respective caudal endplates. No strong correlations were observed between trabecular bone microstructure, maximum endplate concavity depth, vertebral cortex, and endplate thickness parameters. Detailed reference data of these anatomical parameters for each lumbar vertebral body can aid in improved understanding of bone quality, particularly when assessing different implant designs and fixation approaches. Moreover, such anatomical knowledge may help clinicians with optimal implant design selection and surgical placement of these devices into their respective locations.


Antioxidant activity of olive pulp and olive oil phenolic compounds of the arbequina cultivar.
The aim of this study was to characterize antioxidant activities of phenolic compounds that appear in olive pulp and olive oils using both radical scavenging and antioxidant activity tests. Antiradical and antioxidant activities of olive pulp and olive oil phenolic compounds were due mainly to the presence of a 3,4-dihydroxy moiety linked to an aromatic ring, and the effect depended on the polarity of the phenolic compound. Glucosides and more complex phenolics exhibited higher antioxidant activities toward oxidation of liposomes, whereas in bulk lipids aglycons were more potent antioxidants with the exception of oleuropein. Lignans acted as antioxidants only in liposomes, which could partly be due to their chelating activity, because liposome oxidation was initiated by cupric acetate. The antioxidant activity of virgin olive oil is principally due to the dialdehydic form of elenolic acid linked to hydroxytyrosol (3,4-DHPEA-EDA), a secoiridoid derivative (peak RT 36, structure unidentified), and luteolin.


Short-term effect of glucocorticoid injection into the superficial masseter muscle of patients with chronic myalgia: a comparison between fibromyalgia and localized myalgia.
The aim of this study was to investigate whether the treatment effect of intramuscular glucocorticoid injection differs between patients with fibromyalgia and those with localized myalgia of the masseter muscle concerning pain, tenderness to digital palpation, pressure pain threshold, pressure pain tolerance level, maximum voluntary occlusal force, or intramuscular temperature. Twenty-five patients with fibromyalgia and 25 patients with localized myalgia of the masseter muscle were first asked to assess their pain on a visual analogue scale; afterward, a routine clinical examination, including tenderness to digital palpation, was performed. For each patient, the pressure pain threshold, pressure pain tolerance level, and maximum voluntary occlusal force, as well as the intramuscular temperature, were recorded. Finally each patient received an injection of glucocorticoid. The examination and glucocorticoid treatment were repeated after approximately 2 weeks, and a follow-up was performed after another 5 weeks. In the fibromyalgia group, there was a reduced tenderness to digital palpation in response to the treatment. The localized myalgia group responded with a general improvement of symptoms as well as a significant reduction of pain intensity and tenderness to digital palpation. The results of this study indicate that patients with fibromyalgia and localized myalgia in many respects show a similar response to local glucocorticoid treatment.


Basic studies for continuous production of L-aspartic acid by immobilized Escherichia coli cells.
By using a column packed with immobilized Escherichia coli cells entrapped in a polyacrylamide gel lattice, conditions for continuous production of L-aspartic acid from ammonium fumarate were investigated. When a solution of 1 M ammonium fumarate (pH 8.5) containing 1 mM Mg(2+) was passed through the immobilized cell column at a flow rate of space velocity (SV) = 0.8 at 37 C, the highest rate of reaction was attained. From the column effluents, L-aspartic acid was obtained in good yield. The immobilized cell column was very stable.


Oxidoreductase disulfide bond proteins DsbA and DsbB form an active redox pair in Chlamydia trachomatis, a bacterium with disulfide dependent infection and development.
Chlamydia trachomatis is an obligate intracellular bacterium with a distinctive biphasic developmental cycle that alternates between two distinct cell types; the extracellular infectious elementary body (EB) and the intracellular replicating reticulate body (RB). Members of the genus Chlamydia are dependent on the formation and degradation of protein disulfide bonds. Moreover, disulfide cross-linking of EB envelope proteins is critical for the infection phase of the developmental cycle. We have identified in C. trachomatis a homologue of the Disulfide Bond forming membrane protein Escherichia coli (E. coli) DsbB (hereafter named CtDsbB) and-using recombinant purified proteins-demonstrated that it is the redox partner of the previously characterised periplasmic oxidase C. trachomatis Disulfide Bond protein A (CtDsbA). CtDsbA protein was detected in C. trachomatis inclusion vacuoles at 20 h post infection, with more detected at 32 and similar levels at 44 h post infection as the developmental cycle proceeds. As a redox pair, CtDsbA and CtDsbB largely resemble their homologous counterparts in E. coli; CtDsbA is directly oxidised by CtDsbB, in a reaction in which both periplasmic cysteine pairs of CtDsbB are required for complete activity. In our hands, this reaction is slow relative to that observed for E. coli equivalents, although this may reflect a non-native expression system and use of a surrogate quinone cofactor. CtDsbA has a second non-catalytic disulfide bond, which has a small stabilising effect on the protein's thermal stability, but which does not appear to influence the interaction of CtDsbA with its partner protein CtDsbB. Expression of CtDsbA during the RB replicative phase and during RB to EB differentiation coincided with the oxidation of the chlamydial outer membrane complex (COMC). Together with our demonstration of an active redox pairing, our findings suggest a potential role for CtDsbA and CtDsbB in the critical disulfide bond formation step in the highly regulated development cycle.


Sexual behaviour and papillomavirus exposure in cervical intraepithelial neoplasia: a population-based case-control study.
Sexual history is an established risk determinant for cervical neoplasia. It is not clear if human papillomavirus (HPV) exposure entirely explains the sexual behaviour-related risk or if other sexually transmitted agents may act as cofactors for HPV in carcinogenesis. The aim of this study was to elucidate whether HPV exposure or HPV persistence explains the sexual history-related risk of high-grade cervical intraepithelial neoplasia (CIN) using a population-based case-control study of most of the 254 women referred to colposcopy in the Vasterbotten county in Sweden because of an abnormal cervical smear during October 1993 to December 1995 and 320 age-matched women from the general population. The women were interviewed for sexual history and tested for presence of serum antibodies to HPV-16, -18 and -33 as well as for presence of HPV DNA in cervical brush samples. HPV-16, -18 and -33 seropositivity was specific for the corresponding type of HPV DNA, dependent on the lifetime sexual history and associated with a two- to threefold increased risk of CIN 3. There was no sexual history-related risk of CIN among HPV-seropositive women and adjustment for HPV DNA presence explained the sexual history-related risk of CIN. In conclusion, HPV exposure appeared to explain the sexual history-related risk of high-grade CIN.


[Neuroborreliosis].
Neuroborreliosis is easily diagnosed by means of clinical symptoms and laboratory findings. Guiding symptoms are radicular pain and pareses of the extremities and the facial nerve. There is a great number of further less frequently occurring neurological symptoms, which can be attributed to a borrelial infection only by appropriate investigations of the CSF. Radiculitis is cured adequately by oral doxycycline while symptoms of the central nervous system are probably better treated intravenously by ceftriaxone, cefotaxime or penicillin G. Post-Lyme syndrome is a diffuse description of non-specific complaints, which are not the explicit result of a former infection with B. burgdorferi. As further antibiotics do not help and the CSF is unremarkable in most patients, a persistent infection with B. burgdorferi s.l. in all probability can be excluded.


Differential contributions to the transcriptome of duplicated genes in response to abiotic stresses in natural and synthetic polyploids.
Polyploidy has occurred throughout plant evolution and can result in considerable changes to gene expression when it takes place and over evolutionary time. Little is known about the effects of abiotic stress conditions on duplicate gene expression patterns in polyploid plants. We examined the expression patterns of 60 duplicated genes in leaves, roots and cotyledons of allotetraploid Gossypium hirsutum in response to five abiotic stress treatments (heat, cold, drought, high salt and water submersion) using single-strand conformation polymorphism assays, and 20 genes in a synthetic allotetraploid. Over 70% of the genes showed stress-induced changes in the relative expression levels of the duplicates under one or more stress treatments with frequent variability among treatments. Twelve pairs showed opposite changes in expression levels in response to different abiotic stress treatments. Stress-induced expression changes occurred in the synthetic allopolyploid, but there was little correspondence in patterns between the natural and synthetic polyploids. Our results indicate that abiotic stress conditions can have considerable effects on duplicate gene expression in a polyploid, with the effects varying by gene, stress and organ type. Differential expression in response to environmental stresses may be a factor in the preservation of some duplicated genes in polyploids.


Genomic backgrounds of Japanese patients with undiagnosed neurodevelopmental disorders.
Recently, many genes related to neurodevelopmental disorders have been identified by high-throughput genomic analysis; however, a comprehensive understanding of the mechanism underlying neurodevelopmental disorders remains to be established. To further understand these underlying mechanisms, we performed a comprehensive genomic analysis of patients with undiagnosed neurodevelopmental disorders. Genomic analysis using next-generation sequencing with a targeted panel was performed for a total of 133 Japanese patients (male/female, 81/52) with previously undiagnosed neurodevelopmental disorders, including developmental delay (DD), intellectual disability (ID), autism spectrum disorder (ASD), and epilepsy. Genomic copy numbers were also analyzed using the eXome Hidden Markov Model (XHMM). Thirty-nine patients (29.3%) exhibited pathogenic or likely pathogenic findings with single-gene variants or chromosomal aberrations. Among them, 20 patients were presented here. Pathogenic or likely pathogenic variants were identified in 18 genes, including ACTG1, CACNA1A, CHD2, CDKL5, DNMT3A, EHMT1, GABRB3, GABRG2, GRIN2B, KCNQ3, KDM5C, MED13L, SCN2A, SHANK3, SMARCA2, STXBP1, SYNGAP1, and TBL1XR1. A diagnostic yield of 29.3% in this study was nearly the same as that previously reported from other countries. Thus, we suggest that there is no difference in genomic backgrounds in Japanese patients with undiagnosed neurodevelopmental disabilities. Although most of the patients possessed de novo variants, one of the patients showed an X-linked inheritance pattern. As X-linked recessive disorders exhibit the possibility of recurrent occurrence in the family, comprehensive molecular diagnosis is important for genetic counseling.


Renin-angiotensin-aldosterone system inhibition in patients affected by heart failure: efficacy, mechanistic effects and practical use of sacubitril/valsartan. Position Paper of the Italian Society of Cardiology.
Renin-angiotensin-aldosterone system (RAAS) inhibition is a mainstay of the pharmacological treatment of heart failure with reduced ejection fraction (HFrEF). In the last years RAAS blockade has been improved by the introduction of the Angiotensin Receptor-Neprilysin Inhibitor (ARNI) sacubitril/valsartan, that combines RAAS inhibition with the block of neprilysin, boosting the positive effects of natriuretic peptides. The PARADIGM-HF trial demonstrated a significant advantage of sacubitril/valsartan over enalapril on the reduction of cardiovascular (CV) mortality and heart failure hospitalizations rates. Then, several randomized clinical trials and observational studies investigated its role in different clinical settings and its efficacy has been fully recognized in the most recent HFrEF European and USA guidelines. The effects of sacubitril/valsartan on major CV outcomes are associated with reduction of NT-proBNP levels and reverse cardiac remodeling and mitral regurgitation, recognized as one of the mechanistic effects of the drug explaining the favorable prognostic effects. A careful evaluation of patients' clinical profile is relevant to implement the use of ARNI in the clinical practice and to obtain the maximal treatment efficacy. The present Position Paper reports the opinion of the Italian Society of Cardiology on the optimal blockade of the RAAS system in HF patients with the aim of fostering widespread implementation of scientific evidence and practice guidelines in the medical community.


Bridging the gap from research to practice for enhanced health-related quality of life in people with chronic kidney disease.
Improving the health status of people with chronic kidney disease (CKD) through physical activity (PA) or exercise interventions is challenging. One of the gaps in the process of translating the general public PA activity guidelines as well as the CKD-specific guidelines into routine clinical practice is the lack of systematic recording and monitoring of PA and physical function attributes, which can also be used to develop individualized and measurable plans of action to promote PA for health. We aim to present an overview of key considerations for PA, physical function and health-related quality of life (HRQoL) evaluation in people with CKD, with the aim of encouraging health professionals to integrate assessment of these outcomes in routine practices. Physical inactivity and impaired physical function, sometimes to the extent of physical and social disability levels, and subsequently lower perceived HRQoL, are highly prevalent in this population. Enhanced PA is associated with better physical function that also translates into multiple health benefits. Breaking the vicious circle of inactivity and physical dysfunction as early as possible in the disease trajectory may confer huge benefits and enhanced life satisfaction in the longer term. With this in mind, the importance of PA/exercise interventions in CKD to improve HRQoL is also summarized.


Cloning and sequence analysis of the mouse genomic locus encoding the largest subunit of RNA polymerase II.
The genomic locus (RPII215) encoding the largest subunit of mouse RNA polymerase II has been cloned by low stringency hybridization to a Drosophila RPII215 probe. The mouse gene consists of 28 exons which span 30 kilobases. Analysis of the nucleotide and predicted protein sequences indicates that the protein is comprised of two domains. There is a 1500 residue amino-terminal domain which contains seven regions strikingly similar to those in the beta' subunit of Escherichia coli RNA polymerase, and a carboxyl-terminal domain comprised of 52 repeats of a 7-amino-acid consensus sequence Tyr-Ser-Pro-Thr-Ser-Pro-Ser. Among the seven highly conserved regions are a strongly basic domain consistent with a DNA-binding site and a consensus sequence characteristic of a potential zinc-binding domain. The 5' upstream region contains three tandem sequences similar to binding sites for the transcription factor SP1. Two of the introns in this gene splice at donor GC dinucleotides as opposed to previously described invariant GT sites. The identification of regions which are highly conserved as compared with bacterial and yeast RNA polymerase and other regions which are unique to the mouse protein suggests which domains of RNA polymerase large subunits are involved in aspects of transcription common to both procaryotes and eucaryotes and which are characteristic of transcription in higher organisms.


Relationship Between Increased Fucosylation and Metastatic Potential in Colorectal Cancer.
Fucose is utilized for the modification of different molecules involved in blood group determination, immunological reactions, and signal transduction pathways. We have recently reported that enhanced activity of the fucosyltransferase 3 and/or 6 promoted TGF-ß-mediated epithelial mesenchymal transition and was associated with increased metastatic potential of colorectal cancer (CRC), suggesting that fucose is required by CRC cells. With this in mind, we examined requirement of L-fucose in CRC cells and developed fucose-bound nanoparticles as vehicles for delivery of anticancer drugs specific to CRC. In this study, we first examined the expression of fucosylated proteins in 50 cases of CRC by immunochistochemical staining with biotinylated Aleuria aurantia lectin (AAL). Then we carried out an L-fucose uptake assay using three CRC cell lines. Finally, we developed fucose-bound nanoparticles as vehicles for the delivery of an anticancer drug, SN38, and examined tumor growth inhibition in mouse xenograft model (n = 6 mice per group). All statistical tests were two-sided. We found a statistically significant relationship between vascular invasion, clinical stage, and intensity score of AAL staining (P≤ .02). L-fucose uptake assay revealed that L-fucose incorporation, as well as fucosylated protein release, was high in cells rich in fucosylated proteins. L-fucose-bound liposomes effectively delivered Cy5.5 into CRC cells. The excess of L-fucose decreased the efficiency of Cy5.5 uptake through L-fucose-bound liposomes, suggesting an L-fucose receptor dependency. Intravenously injected, L-fucose-bound liposomes carrying SN38 were successfully delivered to CRC cells, mediating efficient tumor growth inhibition (relative tumor growth ratio: no treatment group [NT], 8.29 ± 3.09; SN38-treated group [SN38], 3.53 ± 1.47; liposome-carrying, SN38-treated group [F0], 3.1 ± 1.39; L-fucose-bound, liposome-carrying, SN38-treated group [F50], 0.94 ± 0.89; F50 vs NT,P= .003; F50 vs SN38,P= .02, F50 vs F0,P= .04), as well as prolonging survival of mouse xenograft models (log-rank test,P< .001). Thus, fucose-bound liposomes carrying anticancer drugs provide a new strategy for the treatment of CRC patients.


Growth of the tambaqui Colossoma macropomum (Cuvier) (Characiformes: Characidae): which is the best model?
In order to decide which is the best growth model for the tambaqui Colossoma macropomum Cuvier, 1818, we utilized 249 and 256 length-at-age ring readings in otholiths and scales respectively, for the same sample of individuals. The Schnute model was utilized and it is concluded that the Von Bertalanffy model is the most adequate for these data, because it proved highly stable for the data set, and only slightly sensitive to the initial values of the estimated parameters. The phi values estimated from five different data sources presented a CV = 4.78%. The numerical discrepancies between these values are of not much concern due to the high negative correlation between k and Linfinity viz, so that when one of them increases, the other decreases and the final result in phi remains nearly unchanged.


Factors affecting left ventricular synchronicity in hypertensive patients: are arterial stiffness and central blood pressures influential?
Left ventricular (LV) dyssynchrony is a common finding in patients with hypertension and is associated with LV hypertrophy. Arterial stiffness (AS) and central (aortic) blood pressures play a significant role in end-organ damage such as LV hypertrophy caused by hypertension. The objective of this study was to investigate the relationship between AS, central blood pressures (BP) and LV dyssynchrony. Thirty-five newly diagnosed hypertensive patients and 40 controls were enrolled in the study. The entire study population underwent a comprehensive echocardiographic study including tissue synchrony imaging. The 12 segmental model was used to measure the time to regional peak systolic tissue velocity (Ts) in the LV and two dyssynchrony indices were computed. Parameters of AS including pulse wave velocity (PWV), augmentation index (AIx@75), and central systolic and diastolic BP were evaluated by applanation tonometry. The baseline clinical and echocardiographic parameters of both groups were similar except for their BPs. Dyssynchrony indices were prolonged in patients with hypertension as compared to the controls. The standart deviation of Ts of 12 LV segments in patients with hypertension and the controls were 48.7±18.8 vs. 25.8±13.1, respectively (p<0.001), and the maximal difference in Ts between any 2 of 12 LV segments was 143.9±52.2 for hypertension patients vs. 83.8±39.4 for controls (p<0.001). PWV (11.9±2.5 vs. 9.5±1.4, p<0.001), AIx@75 (27.4±8.3 vs. 18.3±9, p=0.009), and central systolic (147.6±20.8 vs. 105.4±11, p<0.001) and diastolic (99.8±14.4 vs. 72.8±9.5, p<0.001) pressures were higher in patients with hypertension than in the controls, respectively. In multivariable analysis, central systolic BP (ß=0.496, p=0.03), LV mass index (ß=0.232, p=0.027), and body mass index (ß=0.308, p=0.002) were found to be independently related to dyssynchrony. Central systolic BP is an independent predictor of LV dyssynchrony, but Aıx@75 did not have an independent effect on LV synchronicity in patients with newly-diagnosed hypertension.


Impact of humoral immune response on distribution and efficacy of recombinant adeno-associated virus-derived acid alpha-glucosidase in a model of glycogen storage disease type II.
Glycogen storage disease type II (GSDII) is a lysosomal storage disease caused by a deficiency in acid alpha-glucosidase (GAA), and leads to cardiorespiratory failure by the age of 2 years. In this study, we investigate the impact of anti-GAA antibody formation on cross-correction of the heart, diaphragm, and hind-limb muscles from liver-directed delivery of recombinant adeno-associated virus (rAAV)5- and rAAV8-GAA vectors. GAA(-/-) mice receiving 1 x 10(12) vector genomes of rAAV5- or rAAV8-DHBV-hGAA were analyzed for anti-GAA antibody response, GAA levels, glycogen reduction, and contractile function. We demonstrate that restoration of GAA to the affected muscles is dependent on the presence or absence of the antibody response. Immune-tolerant mice had significantly increased enzyme levels in the heart and skeletal muscles, whereas immune-responsive mice had background levels of GAA in all tissues except the diaphragm. The increased levels of activity in immune-tolerant mice correlated with reduced glycogen in the heart and diaphragm and, overall, contractile function of the soleus muscle was significantly improved. These findings highlight the importance of the immune response to rAAV-encoded GAA in correcting GSDII and provide additional understanding of the approach to treatment of GSDII.


[Myxomatous mitral valve degeneration: biochemical aspects and physiopathological considerations].
Mitral valve is a complex structure which is submitted to repeated mechanical constraints. In clinical practice, an increasing incidence of mitral insufficiency resulting from myxomatous degeneration is observed. Since myxomatous degeneration is also observed in defined genetic diseases of connective tissues, we propose the hypothesis that idiopathic mitral insufficiency might result from a minor alteration of the interstitial valvular cells and/or their interactions with their support. After a brief review of the role of the extracellular matrix in the heart, some histopathological and biochemical aspects of myxomatous degeneration are presented. Our data and those of the literature will be summarized and a physiopathological hypothesis proposed for myxomatous mitral valve degeneration.


[Hematogenous osteomyelitis in the adult].
Between 1960 and 1985, 39 cases of haematogenous osteomyelitis in adults have been treated. The clinical signs were somewhat different from those seen in children or adolescents. The difference was due to different patterns in the blood supply of adult bones. Three types are described: acute, sub-acute and suppurating. Some patients purely suffered pain. After the age of 50 the prognosis was much worse. All the cases were treated surgically by evacuation of pus, excision of sequestra and necrosed bone, muscle flaps or bone grafting by the Papineau technique. The results were satisfactory except in the elderly.


Isolation, immortalization, and initial characterization of uterine cell lines: an in vitro model system for the porcine uterus.
The aim of this study was to develop immortalized cell lines from porcine uterus. Endometrial cells including luminal epithelium (LE), glandular epithelium (GE), stroma (ST), and myometrium (MYO) were enzymatically isolated from the uterus of a day 12 pregnant gilt. Primary cultures were immortalized by transduction with a retroviral vector containing the E6 and E7 open reading frames of human papillomavirus type 16 (LXSN-16E6E7) packaged by the amphotropic fibroblast line PA-317. Cells having integrated the vector were selected by resistance to the neomycin analog G418 (0.4-1.5 mg/ml). Surviving cells were maintained in complete culture medium containing G418 (0.1 mg/ml) and subcultured for 1 yr. Expression of the E7 protein was confirmed in all cell lines by Western blotting. Phase contrast microscopy revealed that LE and GE cells exhibited cobblestone morphology, whereas ST and MYO cells exhibited spindle-shaped morphology. The epithelial origin of LE and GE was confirmed by positive immunostaining for cytokeratin. Stromal and MYO cells were vimentin-positive, but cytokeratin-negative. The MYO cell lines were positive for smooth muscle alpha-actin staining, whereas LE, GE, and ST cell lines were negative for alpha-actin. Western blotting indicated that all cell lines expressed both estrogen and progesterone receptors, but only GE cells secreted uteroferrin (UF). Collectively, these porcine uterine cell lines provide an in vitro model for studying cell type-specific actions of hormones and cytokines, signal transduction pathways, cell-cell interactions, and gene expression.


The maize Single myb histone 1 gene, Smh1, belongs to a novel gene family and encodes a protein that binds telomere DNA repeats in vitro.
We screened maize (Zea mays) cDNAs for sequences similar to the single myb-like DNA-binding domain of known telomeric complex proteins. We identified, cloned, and sequenced five full-length cDNAs representing a novel gene family, and we describe the analysis of one of them, the gene Single myb histone 1 (Smh1). The Smh1 gene encodes a small, basic protein with a unique triple motif structure of (a) an N-terminal SANT/myb-like domain of the homeodomain-like superfamily of 3-helical-bundle-fold proteins, (b) a central region with homology to the conserved H1 globular domain found in the linker histones H1/H5, and (c) a coiled-coil domain near the C terminus. The Smh-type genes are plant specific and include a gene family in Arabidopsis and the PcMYB1 gene of parsley (Petroselinum crispum) but are distinct from those (AtTRP1, AtTBP1, and OsRTBP1) recently shown to encode in vitro telomere-repeat DNA-binding activity. The Smh1 gene is expressed in leaf tissue and maps to chromosome 8 (bin 8.05), with a duplicate locus on chromosome 3 (bin 3.09). A recombinant full-length SMH1, rSMH1, was found by band-shift assays to bind double-stranded oligonucleotide probes with at least two internal tandem copies of the maize telomere repeat, TTTAGGG. Point mutations in the telomere repeat residues reduced or abolished the binding, whereas rSMH1 bound nonspecifically to single-stranded DNA probes. The two DNA-binding motifs in SMH proteins may provide a link between sequence recognition and chromatin dynamics and may function at telomeres or other sites in the nucleus.


Human gender differences in an emotional visual oddball task: an event-related potentials study.
Pictures from the Ekman and Friesen series were used in an event-related potentials study to define the timing of occurrence of gender differences in the processing of positive (happy) and negative (fear) facial expressions. Ten male and 10 female volunteers were confronted with a visual oddball design, in which they had to detect, as quickly as possible, deviant happy or fearful faces amongst a train of standard stimuli (neutral faces). Behavioral results suggest that men and women detected fearful faces more quickly than happy ones. The main result is that the N2b component, functionally considered as an attentional orienting mechanism, was delayed in men for happy stimuli as compared with fearful ones. Gender differences observed in the processing of emotional stimuli could then originate at the attentional level of the information processing system.


Analysis of consumer food purchase data used for outbreak investigations, a review.
BackgroundInvestigations of food-borne outbreaks are frequently unsuccessful and new investigation methods should be welcomed. Aim: Describe the use of consumer purchase datasets in outbreak investigations and consider methodological and practical difficulties. Methods: We reviewed published papers describing the use of consumer purchase datasets, where electronic data on the foods that case-patients had purchased before onset of symptoms were obtained and analysed as part of outbreak investigations. Results: For the period 2006-17, scientific articles were found describing 20 outbreak investigations. Most outbreaks involved salmonella or Shiga toxin-producing Escherichia coli and were performed in eight different countries. The consumer purchase datasets were most frequently used to generate hypotheses about the outbreak vehicle where case-interviews had not been fruitful. Secondly, they were used to aid trace-back investigation, where a vehicle was already suspected. A number of methodological as well as (in some countries) legal and practical impediments exist. Conclusions: Several of the outbreaks were unlikely to have been solved without the use of consumer purchase datasets. The method is potentially powerful and with future improved access to big data purchase information, may become a widely applicable tool for outbreak investigations, enabling investigators to quickly find hypotheses and at the same time estimate odds ratios or relative risks hereof. We suggest using the term 'consumer purchase data' to refer to the approach in the future.


Structure of eukaryotic RNA polymerases.
The eukaryotic RNA polymerases Pol I, Pol II, and Pol III are the central multiprotein machines that synthesize ribosomal, messenger, and transfer RNA, respectively. Here we provide a catalog of available structural information for these three enzymes. Most structural data have been accumulated for Pol II and its functional complexes. These studies have provided insights into many aspects of the transcription mechanism, including initiation at promoter DNA, elongation of the mRNA chain, tunability of the polymerase active site, which supports RNA synthesis and cleavage, and the response of Pol II to DNA lesions. Detailed structural studies of Pol I and Pol III were reported recently and showed that the active center region and core enzymes are similar to Pol II and that strong structural differences on the surfaces account for gene class-specific functions.


Pretreatment ADC Values Predict Response to Radiosurgery in Vestibular Schwannomas.
The response rate of vestibular schwannomas to radiation therapy is variable, and there are surgical options available in the event of treatment failure. The aim of this study was to determine whether pre- and posttreatment ADC values can predict the tumor response to radiation therapy. From a data base of 162 patients with vestibular schwannomas who underwent radiation therapy with gamma knife, CyberKnife, or fractionated stereotactic radiation therapy as the first-line therapy between January 2003 and December 2013, we found 20 patients who had pretreatment ADC values. There were 108 patients (including these 20) had serial MR images that included DWI allowing calculated ADC values from 2-132 months after radiation therapy. Two reviewers measured the mean, minimum, and maximum ADC values from elliptical ROIs that included tumor tissue only. Treatment responders were defined as those with a tumor total volume shrinkage of 20% or more after radiation therapy. The pretreatment mean minimum ADC for nonresponders was 986.7 × 10-6 mm2/s (range, 844-1230 × 10-6 mm2/s) and it was 669.2 × 10-6 mm2/s (range, 345-883 × 10-6 mm2/s) for responders. This difference was statistically significant (P < .001). Using a minimum ADC value of 800 × 10-6 mm2/s led to the correct classification of 18/20 patients based on pretreatment ADC values. The intraclass correlation between reviewers was 0.61. No posttreatment ADC values predicted response. Pretreatment ADC values of vestibular schwannomas are lower in responders than nonresponders. Using a minimum ADC value of 800 × 10-6 mm2/s correctly classified 90% of cases.


Association between serum concentrations of 25-hydroxyvitamin D3 and periodontal disease in the US population.
Periodontal disease (PD) is a common chronic inflammatory disease and an important risk factor for tooth loss. Vitamin D might affect periodontal disease risk via an effect on bone mineral density (BMD) or via immunomodulatory effects. The objective was to evaluate whether serum 25-hydroxyvitamin D(3) [25(OH)D(3)] concentrations are associated with PD in the third National Health and Nutrition Examination Survey. We analyzed data on periodontal attachment loss (AL) and serum 25(OH)D(3) concentrations from 11 202 subjects aged > or =20 y. Mean AL was modeled in a multiple linear regression with quintile of serum 25(OH)D(3) concentration as an independent variable. The model was stratified by age and sex and was adjusted for age within age groups, race or ethnicity, smoking, diabetes, poverty income ratio, body mass index, estrogen use, and gingival bleeding. 25(OH)D(3) concentrations were significantly and inversely associated with AL in men and women aged > or =50 y. Compared with men in the highest 25(OH)D(3) quintile, those in the lowest quintile had a mean AL that was 0.39 mm (95% CI: 0.17, 0.60 mm) higher; in women, the difference in AL between the lowest and highest quintiles was 0.26 mm (0.09, 0.43 mm). In men and women younger than 50 y, there was no significant association between 25(OH)D(3) and AL. The BMD of the total femoral region was not associated with AL and did not mediate the association between 25(OH)D(3) and AL. Low serum 25(OH)D(3) concentrations may be associated with PD independently of BMD. Given the high prevalence of PD and vitamin D deficiency, these findings may have important public health implications.


DNA binding of dinuclear iron(II) metallosupramolecular cylinders. DNA unwinding and sequence preference.
[Fe(2)L(3)](4+) (L = C(25)H(20)N(4)) is a synthetic tetracationic supramolecular cylinder (with a triple helical architecture) that targets the major groove of DNA and can bind to DNA Y-shaped junctions. To explore the DNA-binding mode of [Fe(2)L(3)](4+), we examine herein the interactions of pure enantiomers of this cylinder with DNA by biochemical and molecular biology methods. The results have revealed that, in addition to the previously reported bending of DNA, the enantiomers extensively unwind DNA, with the M enantiomer being the more efficient at unwinding, and exhibit preferential binding to regular alternating purine-pyrimidine sequences, with the M enantiomer showing a greater preference. Also, interestingly, the DNA binding of bulky cylinders [Fe(2)(L-CF(3))(3)](4+) and [Fe(2)(L-Ph)(3)](4+) results in no DNA unwinding and also no sequence preference of their DNA binding was observed. The observation of sequence-preference in the binding of these supramolecular cylinders suggests that a concept based on the use of metallosupramolecular cylinders might result in molecular designs that recognize the genetic code in a sequence-dependent manner with a potential ability to affect the processing of the genetic code.


Infrared spectroscopic discrimination between alpha- and 3(10)-helices in globular proteins. Reexamination of Amide I infrared bands of alpha-lactalbumin and their assignment to secondary structures.
We have undertaken a new and more detailed Fourier-transform infrared (FTIR) spectroscopic study of alpha-lactalbumin (in D2O solution) aimed at correlating its secondary structures to observed Amide I' infrared bands. The spectra reported here were interpreted in light of the recently determined crystal structure of alpha-lactalbumin and by comparison with the spectra and structure of the homologous protein lysozyme. Of particular importance is the new evidence supporting the assignment of the band at 1639 cm-1 to 3(10)-helices. This assignment is in excellent agreement with one based on theoretical and experimental studies of 3(10)-helical polypeptides. The frequency observed for 3(10)-helices is distinctly different from that at which alpha-helices are typically found (viz., around 1655 cm-1). In the present study, two bands are clearly resolved in the latter region at 1651 and 1659 cm-1. Both are apparently associated with alpha-helices. These results suggest that for D2O solutions of globular proteins. FTIR spectroscopy can be a facile method for detecting the presence of these two different types of helical conformation and distinguishing between them. This provides a distinct advantage over ultraviolet circular dichroism spectroscopy (UV-CD). This work also provides a basis for future studies of alpha-lactalbumin which examine the effects of environment (e.g., pH, temperature) and ligands (e.g., Ca2+, Mn2+) on its conformation.


Re-evaluation of myocardial FDG uptake in hyperglycemia.
Myocardial [18F] fluorodeoxyglucose (FDG) uptake depends on several metabolic variables in vivo. The effect of different levels of experimentally induced hyperglycemia on myocardial FDG uptake was examined. FDG uptake was studied in young Donryu rats 1 hr after intravenous injection under various pretreatments that increased serum glucose levels. Serum samples were analyzed for glucose, insulin and free fatty acids. Myocardial distribution of FDG was examined with autoradiography. Administration of glucose (n = 42), triiodothyronine (n = 7), epinephrine (n = 7), dehydroascorbic acid (n = 5) and 4 mg streptozotocin (Szt, n = 10) increased glucose levels to 120-200 mg/dl. Dexamethasone (Dex, n = 34) and 6 mg Szt (n = 6) increased glucose levels to 200-450 mg/dl. Myocardial FDG uptake increased proportionately with increases in serum glucose level up to 200 mg/dl. In severe hyperglycemia (serum glucose: 200-450 mg/dl), however, the FDG uptake decreased and did not correlate with blood glucose level. A study of fractional FDG uptake calibrated by the arterial FDG curve confirmed the same results. Heterogeneous distribution of FDG was observed in the myocardium, both in fasting and in severe hyperglycemic conditions. The pattern of FDG uptake by skeletal muscles was similar to that of the myocardium, although the uptake was lower than that in the myocardium. Changes in insulin and free fatty acids levels could not explain the FDG uptake pattern in severe hyperglycemia. Blood FDG uptake level remained constant regardless of glucose level. Hyperglycemia induced a biphasic pattern of myocardial FDG uptake, common with skeletal muscles. The understanding of myocardial FDG uptake characteristics and their dependence on blood glucose is helpful in interpreting myocardial FDG-PET images.


Reversible plasma testosterone levels reduction after gentamicin administration and freund's adjuvant arthritis in rats.
The present study was undertaken in order to investigate the influence of gentamicin on plasma testosterone levels of healthy and with Freund's adjuvant arthritis rats. Gentamicin (40 mg/day for 4 days) induced significant decrease of testosterone levels in comparison with the control group (P<0.025). Intraperitoneal calcium administration (30 mg/ kg bw) prevented gentamicin effect and maintained testosterone levels to that of the control. Decreased testosterone levels were also observed in gentamicin received Freund's adjuvant arthritic rats, in the acute stage of the inflammatory disease (P<0.025), and in the acute stage of Freund's adjuvant arthritis (P<0.001). It is concluded that the administration of gentamicin decreases plasma testosterone levels without any effect on body and seminal vesicles weight. Calcium loading counteracts gentamicin reducing effect on plasma testosterone levels. Freund's adjuvant arthritis influences the function of body and seminal vesicles as it was shown by the reduction of testosterone levels, body and seminal vesicles weight during the acute phase of the inflammatory disease. In any case the effect was reversible.


Analysis of Senescence-Related Differentiation Potentials and Gene Expression Profiles in Human Dental Pulp Stem Cells.
Dental pulp stem cell (DPSC)-mediated dental pulp regeneration is considered a promising method for the treatment of deep caries with pulpitis. However, mesenchymal stem cell (MSC) senescence is an adverse factor from the perspective of cell-based therapies. In this study, we investigated the characteristics and expression profiles of DPSCs from young and old donors. DPSCs from young and old donors were cultured in differentiation medium, and their differentiation potentials were assessed. Long noncoding RNA (LncRNA) microarray assays and a bioinformatic analysis were performed to investigate differences in LncRNA and mRNA expression profiles between DPSCs from young and old donors. We found that DPSCs from young donors exhibited more powerful proliferation ability and greater osteogenic and adipogenic differentiation potentials than DPSCs from old donors. In DPSCs from young donors, numerous LncRNAs were significantly up- (n = 389) or down-regulated (n = 172) compared to DPSCs from old donors. Furthermore, 304 mRNAs were differentially expressed, including 247 up-regulated genes and 57 down-regulated genes in DPSCs from young donors. The bioinformatic analysis identified that several pathways may be associated with DPSC characteristics, such as those involved in the cell cycle and RNA transport, and revealed nuclear transcription factor Y subunit β, general transcription factor IIB, and nuclear receptor subfamily 3 group C member 1 as core regulatory factors and FR249114, FR299091, and ENST00000450004 as core LncRNAs. Our results indicated that senescence impaired the proliferation and differentiation potentials of DPSCs and that donor age is an important factor that affects their use for tooth regeneration. We also provide insight into the mechanisms responsible for senescence in DPSCs.


A cobalt(ii)heteroarylalkenolate precursor for homogeneous Co3O4 coatings by atomic layer deposition.
We present a new and efficient cobalt precursor, CoII(DMOCHCOCF3)2, to prepare Co3O4 thin films and conformal coatings. In the synthesis of this Co complex, heteroaryl moieties and CF3-groups were combined leading to the precursor with high thermal stability and volatility. The suitability of this precursor for ALD deposition was tested on flat silicon substrates and TiO2/C nanofibers upon process optimization. Deposition at 200 °C results in homogeneous and smooth Co3O4 thin films with a growth rate of 0.02 nm per cycle. Conformal coatings have been successfully obtained on TiO2/C nanofibers, making them an attractive platform for surface chemistry studies on high aspect ratio structures for future photocatalysts, sensors, supercapacitors and batteries.


Intravenous anesthetics differentially modulate ligand-gated ion channels.
Heteromeric neuronal nicotinic acetylcholine receptors (nAChRs) are potently inhibited by volatile anesthetics, but it is not known whether they are affected by intravenous anesthetics. Ketamine potentiates gamma-aminobutyric acid type A (GABAA) receptors at high concentrations, but it is unknown whether there is potentiation at clinically relevant concentrations. Information about the effects of intravenous anesthetics with different behavioral profiles on specific ligand-gated ion channels may lead to hypotheses as to which ion channel effect produces a specific anesthetic behavior. A heteromeric nAChR composed of alpha4 and beta4 subunits was expressed heterologously in Xenopus laevis oocytes. Using the two-electrode voltage clamp technique, peak ACh-gated current was measured before and during application of ketamine, etomidate, or thiopental. The response to GABA of alpha1beta2gamma2s GABAA receptors expressed in human embryonic kidney cells and Xenopus oocytes was compared with and without coapplication of ketamine from 1 microm to 10 mm. Ketamine caused potent, concentration-dependent inhibition of the alpha4beta4 nAChR current with an IC50 of 0.24 microm. The inhibition by ketamine was use-dependent; the antagonist was more effective when the channel had been opened by agonist. Ketamine did not modulate the alpha1beta2gamma2s GABAA receptor response in the clinically relevant concentration range. Thiopental caused 27% inhibition of ACh response at its clinical EC50. Etomidate did not modulate the alpha4beta4 nAChR response in the clinically relevant concentration range, although there was inhibition at very high concentrations. The alpha4beta4 nAChR, which is predominantly found in the central nervous system (CNS), is differentially affected by clinically relevant concentrations of intravenous anesthetics. Ketamine, commonly known to be an inhibitor at the N-methyl-D-aspartate receptor, is also a potent inhibitor at a central nAChR. It has little effect on a common CNS GABAA receptor in a clinically relevant concentration range. Interaction between ketamine and specific subtypes of nAChRs in the CNS may result in anesthetic behaviors such as inattention to surgical stimulus and in analgesia. Thiopental causes minor inhibition at the alpha4beta4 nAChR. Modulation of the alpha4beta4 nAChR by etomidate is unlikely to be important in anesthesia practice based on the insensitivity of this receptor to clinically used concentrations.


Modern taurine cattle descended from small number of near-eastern founders.
Archaeozoological and genetic data indicate that taurine cattle were first domesticated from local wild ox (aurochs) in the Near East some 10,500 years ago. However, while modern mitochondrial DNA (mtDNA) variation indicates early Holocene founding event(s), a lack of ancient DNA data from the region of origin, variation in mutation rate estimates, and limited application of appropriate inference methodologies have resulted in uncertainty on the number of animals first domesticated. A large number would be expected if cattle domestication was a technologically straightforward and unexacting region-wide phenomenon, while a smaller number would be consistent with a more complex and challenging process. We report mtDNA sequences from 15 Neolithic to Iron Age Iranian domestic cattle and, in conjunction with modern data, use serial coalescent simulation and approximate Bayesian computation to estimate that around 80 female aurochs were initially domesticated. Such a low number is consistent with archaeological data indicating that initial domestication took place in a restricted area and suggests the process was constrained by the difficulty of sustained managing and breeding of the wild progenitors of domestic cattle.


Highly efficient color-stable deep-blue multilayer PLEDs: preventing PEDOT:PSS-induced interface degradation.
Highly efficient and stable blue light emission is observed in novel copolymers that are produced from specially designed building blocks. A PEDOT:PSS-induced chemical degradation of the polymer light-emitting diodes (PLEDs) is identified at the interface, and it is found to be accompanied by a shift in the emission color. A method to prevent this highly undesirable interaction is presented.


Association of pro-inflammatory cytokines, cortisol and depression in patients with chronic obstructive pulmonary disease.
Evidence suggests that pro-inflammatory cytokines and cortisol play a crucial role in the etiology of chronic obstructive pulmonary disease (COPD) and depression. Depression occurs commonly among COPD patients and an earlier diagnosis would be beneficial. This study investigated the associations between depression, sputum cytokines and salivary cortisol in COPD patients. The diurnal rhythms of sputum IL-1, IL-6, TNF-α and salivary cortisol were measured in COPD patients with depression compared to those only with depression, or COPD and healthy controls. The area under the diurnal variation curves (AUC) over the 24h time course and relative diurnal variation (VAR) were calculated while correlation and regression analysis were performed. Patients with co-morbid depression and COPD showed an increasing sputum IL-1, sputum TNF-α AUC and a decreasing salivary cortisol VAR (P<0.001). The combination of sputum TNF-α AUC, sputum IL-1 AUC, sputum IL-6 AUC and salivary cortisol VAR performed best as a potential biomarker in the diagnosis of depression in COPD patients, with a sensitivity of 94.74% and a specificity of 96.67%. Positive correlations were found between sputum IL-1 AUC and sputum TNF-α AUC versus depressive symptoms, respectively a negative correlation was found between salivary cortisol VAR and depression. They were independently associated with depression in logistic regression models. Depression in COPD is associated with higher 24-h overall levels of sputum IL-1, TNF-α and flattened diurnal salivary cortisol. These non-invasive sputum and salivary biomarkers may serve as a simple clinical tool for the early diagnosis of depression in COPD patients.


Rapid hepatic clearance of full length CCN-2/CTGF: a putative role for LRP1-mediated endocytosis.
CCN-2 (connective tissue growth factor; CTGF) is a key factor in fibrosis. Plasma CCN-2 has biomarker potential in numerous fibrotic disorders, but it is unknown which pathophysiological factors determine plasma CCN-2 levels. The proteolytic amino-terminal fragment of CCN-2 is primarily eliminated by the kidney. Here, we investigated elimination and distribution profiles of full length CCN-2 by intravenous administration of recombinant CCN-2 to rodents. After bolus injection in mice, we observed a large initial distribution volume (454 mL/kg) and a fast initial clearance (120 mL/kg/min). Immunosorbent assay and immunostaining showed that CCN-2 distributed mainly to the liver and was taken up by hepatocytes. Steady state clearance in rats, determined by continuous infusion of CCN-2, was fast (45 mL/kg/min). Renal CCN-2 clearance, determined by arterial and renal vein sampling, accounted for only 12 % of total clearance. Co-infusion of CCN-2 with receptor-associated protein (RAP), an antagonist of LDL-receptor family proteins, showed that RAP prolonged CCN-2 half-life and completely prevented CCN-2 internalization by hepatocytes. This suggests that hepatic uptake of CCN-2 is mediated by a RAP-sensitive mechanism most likely involving LRP1, a member of the LDL-receptor family involved in hepatic clearance of various plasma proteins. Surface plasmon resonance binding studies confirmed that CCN-2 is an LRP1 ligand. Co-infusion of CCN-2 with an excess of the heparan sulphate-binding protamine lowered the large initial distribution volume of CCN-2 by 88 % and reduced interstitial staining of CCN-2, suggesting binding of CCN-2 to heparan sulphate proteoglycans (HSPGs). Protamine did not affect clearance rate, indicating that RAP-sensitive clearance of CCN-2 is HSPG independent. In conclusion, unlike its amino-terminal fragment which is cleared by the kidney, full length CCN-2 is primarily eliminated by the liver via a fast RAP-sensitive, probably LRP1-dependent pathway.


Red blood cell distribution width, multimorbidity, and the risk of death in hospitalized older patients.
We aimed to assess the association between red blood cell distribution width (RDW) and mortality in patients enrolled by a Geriatric Department. One hundred twenty-two patients were followed up during 5 years. The primary end point was all-cause mortality, and hazard ratios were estimated using a Cox proportional hazard model. Higher RDW values were strongly associated with an increased risk of death. Survival curves across RDW quartiles were statistically different according to the log-rank test (p = 0.017). The first quartile presented higher probability of survival compared to the last one. The gradient from lower to higher risk across quartiles was clear both in the 5-year mortality risk and in the mortality rate per 100 person-years, which ranged from 18.9 to 42.6. However, in the Cox regression model after adjusting for age, severity, and other factors, excess risk was only observed in the highest RDW quartile, with a hazard ratio of 2.24 (CI(95%) 1.13-4.42) vs the first quartile. RDW is a good predictor of mortality in hospitalized older adults beyond those with cardiovascular risk factors, and it could serve as an integrative measure of multiple clinical and subclinical processes simultaneously occurring in complex patients.


[The endocrine pancreas. From the isolated islet to the "artificial pancreas" (author's transl)].
The endocrine function of the pancreas consists of the promotion of storage of nutritive substances after meals through the liberation of insulin and to guarantee the mobilization of this food energy through the secretion of glucagon during fasting. Increased hormone production may result from tumors of the islet cells (insulin: insulinoma; glucagon: glucagonoma; gastrin: Zollinger-Ellison syndrome). An absolute or relative insulin deficiency is a characteristic of diabetes mellitus, in which a relative hyperglucagonemia is also of possible pathophysiological significance. This increased secretion of glucagon can be suppressed by somatostatin. While the clinical application of somatostatin in diabetes mellitus seems problematic at present, the use of a glucose-controlled system of insulin infusion ("artificial pancreas") makes possible a metabolic state approaching the healthy condition.


Antifreeze proteins differentially affect model membranes during freezing.
Over the past decade antifreeze proteins from polar fish have been shown either to stabilize or disrupt membrane structure during low temperature and freezing stress. However, there has been no systematic study on how membrane composition affects the interaction of antifreeze proteins with membranes under stress conditions. Therefore, it is not possible at present to predict which antifreeze proteins will protect, and which will damage a particular membrane during chilling or freezing. Here, we analyze the effects of freezing on spinach thylakoid membranes and on model membranes of varying lipid composition in the presence of antifreeze protein type I (AFP I) and specific fractions of antifreeze glycoproteins (AFGP). We find that the addition of galactolipids to phospholipid model membranes changes the effect each protein has on the membrane during freezing. However, the greatest differences observed in this study are between the different types of antifreeze proteins. We find that AFP type I and the largest molecular weight fractions of AFGP induce concentration dependent leakage from, and are fusogenic to the liposomes. This is the first report that an antifreeze protein induces membrane fusion. In contrast, the smallest fraction of AFGP offers a limited degree of protection during freezing and does not induce membrane fusion at concentrations up to 10 mg/ml.


Neuropsychological change in patients with schizophrenia after treatment with quetiapine or haloperidol.
To assess the efficacy of quetiapine, a recently introduced second generation antipsychotic medication, in reducing cognitive impairment in patients with schizophrenia. Prospective, randomized, double-blind clinical trial. 25 patients who met the Diagnostic and Statistical Manual of Mental Disorders, fourth edition, (DSM-IV) criteria for schizophrenia were recruited from 3 Canadian hospitals. After a 48-hour washout period, 25 patients with schizophrenia were randomly assigned to double-blind treatment with quetiapine or haloperidol for 6 months and evaluated with rating scales for psychotic symptoms, mood and extrapyramidal side effects, as well as standardized neuropsychological measures sensitive to 6 cognitive domains: fine motor skill, attention span, verbal reasoning and fluency, visuospatial construction and fluency, executive skills and visuomotor tracking, and immediate recall of verbal and nonverbal materials. The measures were repeated 8 weeks and 6 months after treatment was initiated. Quetiapine improved psychosis and mood without inducing extrapyramidal symptoms. Quetiapine also had beneficial effects on cognitive skills, particularly verbal reasoning and fluency skills and immediate recall, with additional improvements on executive skills and visuomotor tracking and on the average of the 6 cognitive domains with sustained treatment. Patients taking haloperidol showed improvements in general clinical status, but no specific improvements on the positive syndrome, the negative syndrome, depression ratings or cognitive skills. These preliminary results support the potential value of quetiapine for improving cognitive impairment in patients with schizophrenia and emphasize the importance of further research with this promising atypical antipsychotic.


Emulating Target Clinical Trials of Radical Nephrectomy With or Without Lymph Node Dissection for Renal Cell Carcinoma.
To emulate two target clinical trials of radical nephrectomy (RN) with lymph node dissection (LND) vs radical nephrectomy alone. Using the National Cancer Database, we separately emulated an index trial of patients with cT1-3cN0cM0 renal cell carcinoma (RCC), designed to resemble EORTC 30881 ("index trial emulation"), and a hypothetical trial of patients at increased risk for lymph node metastases with cT1-4cN0-1cM0 RCC ("high-risk trial emulation"). A propensity score for LND was estimated using preoperative features (Model 1) or preoperative and pathologic features (Model 2). The associations of LND with overall survival (OS) were estimated using Cox regression with stabilized inverse probability weights. A total of 67,388 patients were included in the index trial emulation. Median follow-up was 49.2 (interquartile range 27.2-74.3) months. LND was not associated with improved OS when adjusting using either Model 1 (hazard ratio [HR] 1.26; 95% confidence interval [CI] 1.20-1.33; P <.0001) or Model 2 (HR 1.13; 95% CI 1.07-1.20; P <.0001). A total of 69,477 patients were included in the high-risk trial emulation. Median follow-up was 48.6 (interquartile range 26.6-73.8) months. LND was not associated with improved OS when adjusting using either Model 1 (HR 1.24; 95% CI 1.18-1.30; P <.0001) or Model 2 (HR 1.09; 95% CI 1.04-1.16; P = .001). In sensitivity analyses, LND was not associated with improved OS across cN stage, pT stage, tumor grade, histologic subtype, or probability of pN1 disease. In observational analyses, that emulate target trials representing EORTC 30881 and a trial of LND in high-risk RCC, LND was not associated with improved OS.


Peer-relationship-problems account for quality of life impairments in pediatric psoriasis.
Most research on HRQoL-impairments in psoriasis has been conducted in adult patients, small pediatric patient samples or samples not representative of the pediatric population at large. We thus aimed to comprehensively describe HRQoL in pediatric psoriasis compared to psoriasis-free children and adolescents, identify domains most commonly affected and analyze its impact on HRQoL while controlling for important other predictors of HRQoL in a representative pediatric sample. The impact of lifetime-prevalence of psoriasis on total and subscale HRQoL was analyzed by complex sample general linear models alone and adjusted for sociodemographic and clinical variables in a population-based sample (n=6518) of children and adolescents aged 11-17. Total HRQoL and the physical domain were significantly affected by lifetime-psoriasis in univariate analysis. In multivariate analyses, lifetime-psoriasis significantly impacted on total HRQoL and the subscale 'quality of relationships with friends/peers'. Although substantial amounts of variance in HRQoL were explained by mental health, independent effects of lifetime-psoriasis remained after adjustment for this covariate. Total explained variance in total HRQoL was 20%. Our findings suggest psoriasis to be a significant burden as it affects HRQoL even when controlling for mental health. Most of this effect appears to be driven by perceived impairments in the quality of relationship with friends/peers. How this exactly occurs needs to be explored in future research. Meanwhile clinicians need to be more attentive to this effect of psoriasis.


Risk factor analysis for sea lice, Caligus rogercresseyi, levels in farmed salmonids in southern Chile.
Sea lice, Caligus rogercresseyi, are ectoparasitic copepods, which severely affect the salmon farming industry in southern Chile, reducing the health status of fish and producing both direct and indirect economic losses. Local farmers have reported increasing infestation levels since 2004, reaching a peak in 2007. In response to this situation, the Chilean Fisheries Service (Sernapesca) developed a surveillance programme; the first step of which consisted of a general survey of salmon farms. This survey included documenting counts of parasite burdens on fish and measurements of several husbandry and environmental factors providing an evaluation of risk factors for the observed infestation levels. The information collected was analysed using a linear mixed model technique, which takes into account the clustered structure of data, decomposing the unexplained variation and assigning it to different aggregation levels of the productive system. Geographical zones, fish species, treatment against sea lice performed 1 month before sampling, stocking density, fish weight and water salinity were the variables significantly associated with sea lice burdens. In contrast, treatments performed 2-3 months before sampling, use of photoperiod in sea cages and water temperature, were not significant. There was significant unexplained variation at all aggregation levels, i.e. sub-zone, fish farm and cage level, with the fish farm level showing the greatest variation.


Faith-based aid, globalisation and the humanitarian frontline: an analysis of Western-based Muslim aid organisations.
This paper focuses on the emergence and modus operandi of Muslim faith-based aid organisations from the West, particularly those from the United Kingdom. Through case studies of Islamic Relief Worldwide and Muslim Hands, it examines the actual and potential added value generated by these humanitarian players in Muslim-majority contexts at times when aid actors from or associated with the West are being perceived by some as instrumental to the political agendas of Western powers, or are being confronted with the consequences thereof. The study analyses Muslim faith-based aid organisations' transnational networks, their implementing partnerships with local faith-based non-governmental organisations (NGOs), and their security position within and their access to insecure contexts, drawing on field examples and opinion from Central Asia, Iraq and Pakistan. It thereby argues that there is ground for an expansion of the role of Muslim aid actors, because of the existence of social and political realities in the field that cannot be always effectively tackled by the dominant international development approaches.


Molecular magnetic resonance imaging of myocardial angiogenesis after acute myocardial infarction.
Angiogenesis is a natural mechanism to restore perfusion to the ischemic myocardium after acute myocardial infarction (MI). Therapeutic angiogenesis is being explored as a novel treatment for MI patients; however, sensitive, noninvasive in vivo measures of therapeutic efficacy are lacking and need to be developed. Here, a molecular magnetic resonance imaging method is presented to noninvasively image angiogenic activity in vivo in a murine model of MI with cyclic Asn-Gly-Arg (cNGR)-labeled paramagnetic quantum dots (pQDs). The tripeptide cNGR homes specifically to CD13, an aminopeptidase that is strongly upregulated during myocardial angiogenesis. Acute MI was induced in male Swiss mice via permanent ligation of the left anterior descending coronary artery. Molecular magnetic resonance imaging was performed 7 days after surgery and up to 2 hours after intravenous contrast agent administration. Injection of cNGR-pQDs resulted in a strong negative contrast that was located mainly in the infarcted myocardium. This negative contrast was significantly less in MI mice injected with unlabeled pQDs and in sham-operated mice injected with cNGR-pQDs. Validation with ex vivo 2-photon laser scanning microscopy revealed a strong colocalization of cNGR-pQDs with vascular endothelial cells, whereas unlabeled pQDs were mostly extravasated and diffused through the tissue. Additionally, 2-photon laser scanning microscopy demonstrated significant microvascular remodeling in the infarct/border zones compared with remote myocardium. cNGR-pQDs allow selective, noninvasive detection of angiogenic activity in the infarcted heart with the use of in vivo molecular magnetic resonance imaging and ex vivo 2-photon laser scanning microscopy.


Species richness, environmental correlates, and spatial scale: a test using South African birds.
Energy and habitat heterogeneity are important correlates of spatial variation in species richness, though few investigations have sought to determine simultaneously their relative influences. Here we use the South African avifauna to examine the extent to which species richness is related to these variables and how these relationships depend on spatial grain. Taking spatial autocorrelation and area effects into account, we find that primary productivity, precipitation, absolute minimum temperature, and, at coarser resolutions, habitat heterogeneity account for most of the variation in species richness. Species richness and productivity are positively related, whereas the relationship between potential evapotranspiration (PET) and richness is unimodal. This is largely because of the constraining effects of low rainfall on productivity in high-PET areas. The increase in the importance of vegetation heterogeneity as an explanatory variable is caused largely by an increase in the range of vegetation heterogeneity included at coarse resolutions and is probably also a result of the positive effects of environmental heterogeneity on species richness. Our findings indicate that species richness is correlated with, and hence likely a function of, several variables, that spatial resolution and extent must be taken into account during investigations of these relationships, and that surrogate measures for productivity should be interpreted cautiously.


CT perfusion parameter values in regions of diffusion abnormalities.
Dynamic CT perfusion imaging is a rapid and widely available method for assessing cerebral hemodynamics in the setting of ischemia. Nevertheless, little is known about perfusion parameters within regions of diffusion abnormality. Since MR diffusion-weighted (DW) imaging is widely considered the most sensitive and specific technique to examine the ischemic core, new knowledge about CT perfusion findings in areas of abnormal diffusion would likely provide valuable information. The purpose of our study was to measure the CT-derived perfusion values within acute ischemic lesions characterized by 1) increased signal intensity on DW images and 2) decreased apparent diffusion coefficient (ADC) and compare these values with those measured in contralateral, normal brain tissue. Analysis was performed in 10 patients with acute middle cerebral artery territory stroke of symptom onset less than 8 hours before imaging who had undergone both CT perfusion and DW imaging within 2 hours. After registration of CT perfusion and DW images, measurements were made on a pixel-by-pixel basis in regions of abnormal hyperintensity on DW images and in areas of decreased ADC. Significant decreases in cerebral blood flow and cerebral blood volume with elevated mean transit times were observed in regions of infarct as defined by increased signal intensity on DW images and decreased ADC. Comparison of perfusion parameters in regions of core infarct differed significantly from those measured in contralateral normal brain. CT perfusion findings of decreased cerebral blood flow, mean transit time, and cerebrovascular volume correlate with areas of abnormal hyperintensity on DW images and regions of decreased ADC. These findings provide important information about perfusion changes in acute ischemia in areas of diffusion abnormality.


All-Epiphyseal ACL Reconstruction in Children: Review of Safety and Early Complications.
All-epiphyseal anterior cruciate ligament (ACL) reconstruction is a well-described technique for skeletally immature patients. The purpose of this study was to elucidate the early complication rate and identify associated risk factors for rerupture after this procedure in children. We retrospectively reviewed patients who underwent all-epiphyseal ACL reconstructions performed at a large, tertiary care children's hospital between January 2007 and April 2013. Relevant postoperative data including the development of leg-length discrepancy, angular deformity, rerupture, infection, knee range of motion, arthrofibrosis, and other complications were recorded. Independent variables analyzed for association with rerupture included age, body mass index, graft type, graft size, and associated injuries addressed at surgery. A total of 103 patients (average 12.1 y old; range, 6.3 to 15.7) were analyzed. The mean follow-up was 21 months. The overall complication rate was 16.5% (17/103), including 11 reruptures (10.7%), 1 case (<1.0%) of clinical leg-length discrepancy of <1 cm, and 2 cases (1.9%) of arthrofibrosis requiring manipulation under anesthesia. Two patients (1.9%) sustained contralateral ACL ruptures and 3 (2.9%) sustained subsequent ipsilateral meniscus tears during the study period. There were no associations found between age, sex, graft type, graft thickness, body mass index, or associated injuries addressed during surgery and rerupture rate. Knee flexion continued to improve by 20 degrees on an average between the 6 weeks and 6 months postoperative visits (P<0.001; paired samples Student's t test). When taken in the context of known risk of future injury in an ACL-deficient knee, all-epiphyseal ACL reconstruction in children is safe. The rate of growth disturbance in this study is similar to previous reports in this patient demographic. The rerupture rate in this cohort is slightly higher compared with ACL reconstruction in older patients. Level IV-retrospective case series.


Intravitreal Injection of Normal Saline Induces Retinal Degeneration in the C57BL/6J Mouse.
To investigate the adverse effect of intravitreal injection of normal saline (NS) and phosphate buffered saline (PBS) in mouse eyes. NS or PBS was injected intravitreally into C57BL/6J mouse eyes. Retinal lesions were monitored by fundus imaging, spectral-domain optical coherence tomography (SD-OCT), and histological investigations. Retinal immune gene expression was determined by real-time polymerase chain reaction (PCR). The toxic effect of NS and PBS or retinal protein from NS- or PBS-injected eyes on retinal pigment epithelium (RPE) was tested in B6-RPE-07 mouse RPE cell cultures. Intravitreal injection of NS dose-dependently induced localized retinal lesion in mice. Histological investigations revealed multiple vacuoles in photoreceptor outer segments and RPE cells. The lesions recovered over time and by 3 weeks post injection the majority of lesions vanished in eyes receiving 1 μl NS. Inflammatory genes, including TNF-α, IL-1β, IL-6, iNOS, and VEGF were upregulated in NS injected eyes. Intravitreal injection of PBS did not cause any pathology. The treatment of B6-RPE07 cells with 30% PBS or 30% NS did not affect RPE viability. However, incubation of 1-μg/ml retinal protein from NS-injected eyes, but not PBS-injected eyes induced RPE cell death. NS is toxic to the C57BL/6J mouse retina and should not be used as a vehicle for intraocular injection. PBS is not toxic to the retina and is a preferred vehicle. NS is not a physiological solution for intraocular injection in the C57BL/6J mice and questions its suitability for intraocular injection in other species, including human.


Intrinsic disorder and protein multibinding in domain, terminal, and linker regions.
Intrinsic disorder is believed to contribute to the ability of some proteins to interact with multiple partners which is important for protein functional promiscuity and regulation of the cross-talk between pathways. To better understand the mechanisms of molecular recognition through disordered regions, here, we systematically investigate the coupling between disorder and binding within domain families in a structure interaction network and in terminal and inter-domain linker regions. We showed that the canonical domain-domain interaction model should take into account contributions of N- and C-termini and inter-domain linkers, which may form all or part of the binding interfaces. For the majority of proteins, binding interfaces on domain and terminal regions were predicted to be less disordered than non-interface regions. Analysis of all domain families revealed several exceptions, such as kinases, DNA/RNA binding proteins, certain enzymes, and regulatory proteins, which are candidates for disorder-to-order transitions that can occur upon binding. Domain interfaces that bind single or multiple partners do not exhibit significant difference in disorder content if normalized by the number of interactions. In general, protein families with more diverse interactions exhibit less average disorder over all members of the family. Our results shed light on recent controversies regarding the relationship between disorder and binding of multiple partners at common interfaces. In particular, they support the hypothesis that protein domains with many interacting partners should have a pleiotropic effect on functional pathways and consequently might be more constrained in evolution.


Convergent iterative closest-point algorithm to accomodate anisotropic and inhomogenous localization error.
Since its introduction in the early 1990s, the Iterative Closest Point (ICP) algorithm has become one of the most well-known methods for geometric alignment of 3D models. Given two roughly aligned shapes represented by two point sets, the algorithm iteratively establishes point correspondences given the current alignment of the data and computes a rigid transformation accordingly. From a statistical point of view, however, it implicitly assumes that the points are observed with isotropic Gaussian noise. In this paper, we show that this assumption may lead to errors and generalize the ICP such that it can account for anisotropic and inhomogenous localization errors. We 1) provide a formal description of the algorithm, 2) extend it to registration of partially overlapping surfaces, 3) prove its convergence, 4) derive the required covariance matrices for a set of selected applications, and 5) present means for optimizing the runtime. An evaluation on publicly available surface meshes as well as on a set of meshes extracted from medical imaging data shows a dramatic increase in accuracy compared to the original ICP, especially in the case of partial surface registration. As point-based surface registration is a central component in various applications, the potential impact of the proposed method is high.


Helplessness and loss as mediators between pain and depressive symptoms in fibromyalgia.
This study evaluated the contribution of condition-specific helplessness and loss to depression in fibromyalgia (FM). Two models were tested. The first model examined whether loss, measured by the West Haven-Yale Multidimensional Pain Inventory (WHYMPI) Interference Scale, would mediate the relationship between disability and depression. The second model determined whether condition-specific helplessness and loss would mediate the relationship between pain and depression with disability controlled. Eighty patients with confirmed diagnoses of FM were recruited throughout Southern California from general medical clinics, newspaper advertisements, and rheumatology practices. The study design was cross-sectional, using self-report, observational, and interview measures. A composite measure of depression was adopted, consisting of the Center for Epidemiological Studies-Depression Scale and the Hamilton Rating Scale for Depression. Hierarchical multiple regression analyses were conducted using a path analytic framework to examine each model. In Model 1, loss fully mediated the relationship between disability and depression. In Model 2, condition-specific helplessness mediated the relationship between pain and depression, but the contribution of loss was not significant. The findings confirm the importance of helplessness and demonstrate that the cognitive meaning of having FM plays a more central role in predicting depressive symptomatology than illness-related stressors, such as pain or disability.


[Diagnosis of sphincter of Oddi dysfunction in patients with postcholecystectomy syndrome from hepatobiliary scintigraphic findings].
to diagnose and estimate the clinical value of postcholecystectomy sphincter of Oddi dysfunction in patients. Examinations were made in 100 postcholecystectomy patients without signs of cholestasis; of them 14 postpapillotomy patients formed a comparison group. Hepatobiliary scintigraphy using the radiotracer 99mTC-bromeside was performed for 90 minutes with cholagogue breakfast at 45 minutes. Common bile duct and duodenal functions and duodenogastric reflux (DGR) were evaluated comparing them with clinical, laboratory, and instrumental findings. Two patient groups were identified according to bile outflow changes. In Group I consisting of 20 (23.2%) patients, the time of maximum accumulation (Tmax) of the radiopharmaceutical in the projection of the choledochus coincided with that in the cholagogue test (46.0 1.8 min) and in Group 2 including 66 (76.8%) patients that was shorter than in the cholagogue test (32.9 +/- 6.8 min) (p<0.05). In Group 2, Tmax was similar to that in the comparison group (30.9 +/- 7.5 min; p > 0.05) and there was no significant difference in intestinal imaging time (18.6 +/- 6.0 min versus 17.6 +/- 0.8) either, which could be indicative of sphincter of Oddi dysfunction. Diarrhea was observed in 73% of the patients with sphincter of Oddi dysfunction and in 86% of the patients in the comparison group versus 10% of the patients with normal bile passage (p<0.01). Statistical data processing showed a correlation of the indicators of sphincter of Oddi dysfunction with those of duodenal evacuator function (r = 0.57; p < 0.0005) and DGR (r = 0.74; p < 0.009). Postcholecystectomy sphincter of Oddi dysfunction assumes the greatest clinical value in patients with duodenal motor-evacuator dysfunction, which should be hepatobiliamy scintigraphic, kept in mind when choossphincter of Oddi dysfunction ing a treatment policy.


High-fat diet alters N-glycosylation of PTPRJ in murine liver.
Protein tyrosine phosphatases (PTPs) regulate multiple signaling pathways. Disruption of tyrosine phosphorylation through imbalanced action between protein tyrosine kinases (RTKs) and PTPs is a hallmark of metabolic disorders, including insulin resistance. A representative member of the receptor-type PTP family, PTPRJ (DEP-1), was previously identified as a negative regulator of insulin signaling and possesses post-translational glycosylation sites. In this regard, it seems of great importance to decipher the structure of PTPRJ's glycosylation, particularly in the context of metabolic disturbances, but this has not been done in detail. Thus, here we aimed at characterizing the glycosylation pattern of PTPRJ in liver. We show that N-glycosylation accounts for up to half of PTPRJ's molecular weight. Applying mass spectrometry, we detected increased levels of high-mannose structures in PTPRJ in liver tissue of obese mice compared to lean littermates. In addition, complex neutral structures without fucose were also elevated in PTPRJ of high-fat diet (HFD) mice. Conversely, complex fucosylated N-glycans as well as sialylated bi- and triantennary N-glycans, were significantly reduced in PTPRJ of HFD-derived liver tissue compared to LFD by ∼two fold (P≤.01, P≤.0001 and P≤.001, respectively). In congruence with these findings, the mannosidase MAN2A1, responsible for the conversion of high-mannose to complex N-glycans, was significantly downregulated under HFD conditions. Here we present for the first time that HFD-induced obesity impacts on the glycosylation pattern of the insulin signaling component PTPRJ in liver. These findings may inspire new research on the glycosylation of PTPs in metabolic diseases and may open up new therapeutic approaches.


Molecular pathogenesis of Friedreich ataxia.
Friedreich ataxia, the most common type of inherited ataxia, is itself caused in most cases by a large expansion of an intronic GAA repeat, resulting in decreased expression of the target frataxin gene. The autosomal recessive inheritance of the disease gives this triplet repeat mutation some unique features of natural history and evolution. Frataxin is a mitochondrial protein that has homologues in yeast and even in gram-negative bacteria. Yeast organisms deficient in the frataxin homologue accumulate iron in mitochondria and show increased sensitivity to oxidative stress. This suggests that Friedreich ataxia is caused by mitochondrial dysfunction and free radical toxicity.


Identification of types II, IX and X collagens at the insertion site of the bovine achilles tendon.
Achilles tendinous collagen fibrils insert into the calcaneus by first passing through a zone that is defined histologically as fibrocartilaginous. This zone consists of four regions: tendon proper, non-mineralized and mineralized fibrocartilage and bone. The function of this zone has not yet been clearly defined. To gain more insight into the role of this fibrocartilaginous zone, collagens present in the zone of the Achilles tendon-calcaneus interface were isolated and characterized. Types II, IX and X collagens were identified in the pepsin digests of the tissue harvested from the bovine Achilles tendon-calcaneus interface. Western blotting using specific antisera to types II, IX and X collagens confirmed the identity of these collagens. Immunofluorescence localization placed type X collagen predominantly in the mineralized zone of the tendon-calcaneus junction, while type IX collagen was distributed throughout the the insertion site. The presence of the cartilage-specific collagens at the Achilles tendon-calcaneus-interface suggests that this zone is cartilaginous in nature. The presence of type X collagen at this junction is not clear, but our present findings go along with the previous report which showed that type X collagen is present in the mineralized zone of the medial collateral ligament femoral insertion site. These data suggest that type X collagen may be a resident of mineralized fibrocartilaginous zones of tendon or ligament-bone junctions and may participate in anchoring ligament or tendon to bone.


Antibacterial activities of the methanol extracts of ten Cameroonian vegetables against Gram-negative multidrug-resistant bacteria.
Many edible plants are used in Cameroon since ancient time to control microbial infections. This study was designed at evaluating the antibacterial activities of the methanol extracts of ten Cameroonian vegetables against a panel of twenty nine Gram negative bacteria including multi-drug resistant (MDR) strains. The broth microdilution method was used to determine the Minimal Inhibitory Concentrations (MIC) and the Minimal Bactericidal Concentrations (MBC) of the studied extracts. When chloramphenicol was used as a reference antibiotic, the MICs were also determined in the presence of Phenylalanine-Arginine β-Naphtylamide (PAβN), an efflux pumps inhibitor (EPI). The phytochemical screening of the extracts was performed using standard methods. All tested extracts exhibited antibacterial activities, with the MIC values varying from 128 to 1024 mg/L. The studied extracts showed large spectra of action, those from L. sativa, S. edule, C. pepo and S. nigrum being active on all the 29 bacterial strains tested meanwhile those from Amaranthus hybridus, Vernonia hymenolepsis, Lactuca.carpensis and Manihot esculenta were active on 96.55% of the strains used. The plant extracts were assessed for the presence of large classes of secondary metabolites: alkaloids, anthocyanins, anthraquinones, flavonoids, phenols, saponins, steroids, tannins and triterpenes. Each studied plant extract was found to contain compounds belonging to at least two of the above mentioned classes. These results confirm the traditional claims and provide promising baseline information for the potential use of the tested vegetables in the fight against bacterial infections involving MDR phenotypes.


Cns tumors at AKU: an update plus a brief discussion on intraventricular tumors with special emphasis on central neurocytoma.
This paper is intended to be an update of our earlier published work on CNS tumors along with additional information on intraventricular tumors. Three cases of central neurocytoma are also discussed. An analysis conducted in the Section of Histopathology, AKU to determine the frequency of all CNS neoplasms diagnosed between Jan 1, 1994 and Dec 31, 2001. Frequency of intraventricular tumors was also determined. Histological characterization of the tumors was based on the WHO and AFIP (USA) classification systems for CNS tumors. A total of 1677 CNS tumors were diagnosed during the study period. 1510(90.04%) were primary, and 167(9.96%) were metastatic. Among the primary tumors, gliomas constituted the largest category with 825 cases or 54.63% of all primary tumors. Meningothelial tumors comprised the second largest group with 364 cases or 24.10%. among other major categories, embayonal tumors, and peripheral nerve sheath tumors comprised 6.75% and 6.82% of all primary tumors. Among less common tumors, Non-Hodgkin's lymphomas and hemangioblastomas comprised 3.11% and 1.52% of all CNS tumors. Intraventricular tumors comprised 7.41% of all primary tumors. Ependymomas comprised 64.28% of all Intraventricular tumors. Gliomas and meningothelial tumors are the commonest group of primary CNS tumors. Metastatic tumors are quite common.


Duration of Postoperative Fever as a Simple and Useful Prognostic Indicator in Gastric Cancer Patients.
Postoperative inflammation is associated with cancer progression in several cancers. However, the prognostic significance of postoperative fever remains unclear in gastric cancer patients. We enrolled 442 patients with a histopathological diagnosis of gastric adenocarcinoma who underwent curative surgery. The mean duration of postoperative fever ≥ 37°C was 8.7 days (range: 0-186 days) and significantly longer in patients with advanced gastric cancer, venous invasion, and open or total gastrectomy vs. patients with early gastric cancer (P = 0.0072), no venous invasion (P = 0.025), laparoscopic gastrectomy (P = 0.027), and either proximal or distal partial gastrectomy (P = 0.0015). Five-year overall survival rates were 69.5% vs. 83.6% in the prolonged postoperative fever group (≥ 6 days of ≥ 37°C) vs. the nonprolonged group (< 6 days of ≥ 37°C), respectively (P = 0.0008). In patients without Clavien-Dindo classification postoperative infectious complications grade ≥ 2, 5-year overall survival was 69.7% vs. 84.0% in patients with prolonged postoperative fever vs. those without, respectively (P = 0.0067). Five-year disease-specific survival was 85.9% vs. 93.1% in patients with prolonged fever vs. those without, respectively (P = 0.041). Multivariate analysis indicated that postoperative fever was an independent prognostic indicator. Postoperative fever ≥ 37°C duration may be useful in predicting prognosis in gastric cancer patients.


Spontaneous formation of a nonuniform chiral spin liquid in a moat-band lattice.
A number of lattices exhibit moatlike band structures, i.e., a band with infinitely degenerate energy minima attained along a closed line in the Brillouin zone. If such a lattice is populated with hard-core bosons, the degeneracy prevents their condensation. At half-filling, the system is equivalent to the s=1/2 XY model at a zero magnetic field, while the absence of condensation translates into the absence of magnetic order in the XY plane. Here, we show that the ground state breaks time reversal as well as inversion symmetries. This state, which may be identified with the chiral spin liquid, has a bulk gap and chiral gapless edge excitations. The applications of the developed analytical theory include an explanation of recent numerical findings and a suggestion for the chiral spin liquid realizations in experiments with cold atoms in optical lattices.