Very early activation of m-calpain in peripheral nerve during Wallerian degeneration.
Peripheral nerve injury results in a series of events culminating in degradation of the axonal cytoskeleton (Wallerian degeneration). In the time period between axotomy and cytoskeletal degradation (24-48 h in rodents), there is calcium entry and activation of calpains within the axon. The precise timing of these events during this period is unknown. In the present study, antibodies were generated to three distinct peptide epitopes of m-calpain, and a fusion protein antibody was generated to the intrinsic calpain inhibitor calpastatin. These antibodies were used to measure changes in these proteins in mouse sciatic nerves during Wallerian degeneration. In sciatic nerve homogenates and cultured dorsal root ganglion (DRG) neurites, m-calpain protein was significantly reduced in transected nerves very early after nerve injury, long before axonal degeneration occurred. Levels of m-calpain protein remained low as compared to control nerves for the remainder of the 72-h time course. No changes in calpastatin protein were evident. Systemic treatment of animals with the protease inhibitor leupeptin partially prevented the rapid loss of calpain protein. Removal of calcium in DRG cultures had the same effect. These data indicate that m-calpain protein is lost very early after axonal injury, and likely reflect activation and degradation of this protein long before the cytoskeleton is degraded. Calpain activation may be an early event in a proteolytic cascade that is initiated by axonal injury and culminates with axonal degeneration.


Free / total prostate specific antigen ratio for prostate cancer detection: a prospective blind study.
We conducted this prospective study to evaluate the ability of the percentage of free PSA (% fit PSA) to improve the specificity of prostate cancer (PC) detection. Seven hundred and eighty-five patients with serum PSA levels between 4 and 25 ng/ml underwent sextant biopsies of the prostate. The % f/t PSA was analyzed for the sensitivity, specificity and positive predictive values. The mean % f/t PSA of the 169 patients with PC was 18.0 +/- 7.7 and was significantly (p < 0.0001) lower than that of the 616 patients who had histologically benign biopsy specimens, with a mean % f/t PSA of 26.6 +/- 9.7. Using 30% f/t PSA as the cut-off eliminated 34.4 % of the negative biopsies while still detecting 93.4% of the carcinomas. The optimal cut-off value of 30% f/t PSA may be applied as reference using the Cis bio international assay for PC detection.


Emblica officinalis and hepatocarcinogenesis: a chemopreventive study in Wistar rats.
Emblica officinalis L. commonly known as gooseberry, Phyllanthus emblica, Emblica, Indian gooseberry, amla is used in Indian system of medicine for the treatment of liver ailments. In the present study, we report that it ameliorates the carcinogenic response as it reverses the histopathological changes and reduces the number of gamma-GT-positive foci induced by Solt Farber protocol in the liver of Wistar rats. Tumors were induced by initiation with diethylnitrosoamine (DEN) (200 mg/kg body wt., i.p.) followed by promotion with 2-acetylaminoflourine (2-AAF) (0.02%, w/w in diet) for continuous 6 weeks and the animals were subjected to partial hepatectomy on day 21 of initiation. Pretreatment with defatted methanolic fruit extract (100 and 200mg/kg b.w.) of Emblica officinalis showed significant partial recovery of pathological manifestations as compared to DEN and 2-AAF-treated group animals (p<0.001) and suppressed the tumor forming potential of 2-AAF (p<0.05) at both the doses. Emblica officinalis has the potential to suppress carcinogen-induced response in rat liver.


Recommendations to improve end-of-life care through regulatory change in U.S. health care financing.
Palliative care has emerged as the interdisciplinary approach to relieving suffering and improving quality of life. Unfortunately, it is not yet available to all who need it. Although legislation may ultimately be needed, regulatory changes can quickly improve the current situation. While we see many regulatory changes that are possible, just three would quickly improve care: adjust hospital payment to include essential palliative care services for hospitalized patients in pertinent Diagnostic Related Groups (DRGs); clarify physician payment for providing palliative care by clarifying guidelines for carriers and paying for case management services; and clarify the Medicare Hospice Benefit by defining the 6 month prognosis in statistical terms.


The relation of NCCR variations and host transcription factors gene regulation in BK polyomavirus infected kidney transplant patients.
BK polyomavirus (BKPyV) infection in immunocompromised patients can led to polyomavirus-associated nephropathy (BKPyVAN) especially after kidney transplantation. The polyomavirus genome contains enhancer elements that are important transcription activators. In this study, the association between viral and host gene expression and NCCR variations was evaluated in kidney transplant recipients (KTRs) with BKPyV active, and BKPyV in-active infection. Blood samples were collected from selected KTRs who divided to patients with active and in-active BKPyV infection. Transcriptional control region (TCR) anatomy was compared to the genomic sequence of archetype BKPyV strain WW using nested PCR method and sequencing. The expression level of some transcription factor genes was evaluated using in-house Real-time PCR (SYBR Green) technique. Most changes were observed after TCR anatomy detection in the Q and P blocks. The expression level of VP1 and LT-Ag viral genes were significantly higher in patients with active infection compared with non-infected ones. Transcription factor genes SP1, NF1, SMAD, NFκB, P53, PEA3, ETS1, AP2, NFAT and AP1 were significantly higher in BKPyV active group in comparison in-active and control groups. The analyses revealed that viral load level and mutations frequency has significant correlation. Based on the results, increasing of NCCR variations were associated with higher viral load of BKPyV especially in Q block. Host transcriptional factors and viral genes all had higher express level in active BKPyV patients versus no in-active ones. Detection of the relation between NCCR variation and BKPyV severity in KTRs need to be confirmed in further complicated studies.


Plasmon-gating photoluminescence in graphene/GeSi quantum dots hybrid structures.
The ability to control light-matter interaction is central to several potential applications in lasing, sensing, and communication. Graphene plasmons provide a way of strongly enhancing the interaction and realizing ultrathin optoelectronic devices. Here, we find that photoluminescence (PL) intensities of the graphene/GeSi quantum dots hybrid structures are saturated and quenched under positive and negative voltages at the excitation of 325 nm, respectively. A mechanism called plasmon-gating effect is proposed to reveal the PL dependence of the hybrid structures on the external electric field. On the contrary, the PL intensities at the excitation of 405 and 795 nm of the hybrid structures are quenched due to the charge transfer by tuning the Fermi level of graphene or the blocking of the excitons recombination by excitons separation effect. The results also provide an evidence for the charge transfer mechanism. The plasmon gating effect on the PL provides a new way to control the optical properties of graphene/QD hybrid structures.


Phthalimidomethyl as a drug pro-moiety. Probing its reactivity.
Phthalimidomethyl derivatives 1, encompassing a wide range of leaving group abilities, are rapidly hydrolysed to the corresponding phthalamic acid via rate-determining attack at the phthalimide carbonyl group.


Failure to induce inhibition of cyclin A and up-regulation of p21 expression in phorbol ester-resistant U937 cells by phorbol ester.
Differentiation resistant U937 cells were derived from parental U937 cells by selecting for continuously growing U937 cells in cell cultures continuously exposed to phorbol 12 myristate 13-acetate (PMA). Unlike in other known PMA resistant U937, the basal expression of protein kinase C (PKC) isozymes in these PMA resistant cells (R-U937) was significantly decreased. Subsequent analyses revealed differences between the wild type U937 and the R-U937 cells with respect to G1 phase arrest, which seemed to occur in U937 because of low levels of cdk2 kinase activity. This abolished cdk2 kinase activity is mainly due to inhibition of cdk2 phosphorylation, cyclin A down-regulation and cyclin dependent kinase inhibitor p21 up-regulation. Our data suggest that events down-stream of PKC activation may mediate cell cycle control. Thus, the R-U937 cells could be useful for further PKC mediated cell cycle control studies.


Optical rotation signal of nerve and the Faraday effect.
The polarity of the optical rotation signal, recorded from the lobster nerve, was reversed when the direction of incident light was changed by 180 degrees. This is a property of the Faraday rotation, but the magnetic field by the action current was too small to explain the size of the observed signal.


Curcumin induces osteosarcoma MG63 cells apoptosis via ROS/Cyto-C/Caspase-3 pathway.
The antitumor effects of curcumin have attracted widespread attention worldwide. One of its major functions is to induce the apoptosis of tumor cells, but the antitumor mechanism is currently unclear. In the present study, we found that cell mortality and curcumin concentration were dose dependent. Curcumin of low concentrations (10 μΜ) could reduce the level of reactive oxygen species (ROS) in tumor cells, while curcumin of high concentrations (80 μΜ) was able to significantly increase the content of ROS. In addition, Western blotting detection suggested that curcumin of high concentrations can induce the release of Cyto-C and the activation of Caspase-3, and that ROS scavenger NAC apparently inhibits apoptosis protein release and activation, consequently slowing the curcumin-induced apoptosis. Taken together, curcumin further activates the mitochondrial apoptotic pathway by inducing cells to generate ROS and ultimately promotes the apoptosis of tumor cells.


Neutrophil priming: the cellular signals that say 'amber' but not 'green'.
One of the most intriguing gaps in our understanding of how neutrophils work concerns the mechanism by which the oxidase response in these cells is 'primed'. In the primed state, there is no increase in oxidase activity, yet subsequent stimulation provokes a response that is larger than in nonprimed, activated cells. Thus, neutrophils exist in one of three states: quiescent, primed or active. Individual primed cells may be thought of as being 'ready to go' but awaiting further stimulus before the oxidase response is elicited. The primed neutrophils are thus held at 'amber', awaiting 'green' before activity is triggered. Here, Maurice Hallett and Darren Lloyds suggest a molecular basis for the signals that say 'amber' but not 'green'.


Inhibition of baroreceptor and chemoreceptor reflexes on heart rate by afferents from the lungs.
1. Brief stimuli were delivered to the carotid baroreceptors or chemoreceptors in dogs anaesthetized with chloralose and paralysed with D-turbocurarine. Baroreceptor stimulation was achieved by forceful retrograde injection of 2-5 ml or air-equilibrated saline into the external carotid artery after first clamping the common carotid artery. Chemoreceptor stimulation was achieved by rapid retrograde injections of 0.2-0.5 ml of warmed CO2-equilibrated saline into the external carotid artery. Observations were made during periods of temporary cessation of artificial ventilation. 2. When the volume of the lungs was not changing, prompt decreases in heart rate were evoked by chemoreceptor or baroreceptor stimuli except when these were delivered during the inspiratory phase of breathing (as judged from the records of phrenic nerve activity). 3. No changes in heart rate were evoked when these stimuli were timed to occur during expansion of the lungs in response to a rapid increase in intratracheal pressure (6-10 mmHg in 1-2 sec). Decreases in heart rate were evoked when the stimuli occurred during slower inflations of the lungs. 4. Both stimuli regained their effectiveness on heart rate with time after inflations of the lung when the lungs were held inflated. Both stimuli evoked large decreases in heart rate when delivered during deflations of the lung. 5. The effects of lung inflation on the effectiveness of both cardiodepressor reflexes were abolished by surgical denervation of the lungs.


Aortic valve insufficiency and coronary artery narrowing in a child with polyarticular juvenile rheumatoid arthritis.
Involvement of the cardiac valves in juvenile rheumatoid arthritis (JRA) is a rare but life threatening complication. We report a 9 1/2-year-old girl with rheumatoid factor positive polyarticular JRA who developed aortic insufficiency that required valve replacement. Six months later she developed angina due to narrowing of the left coronary artery, probably secondary to aortitis.


Adsorption of l-buthionine sulfoximine on Bi(III) and Eu(III) co-substituted hydroxyapatite nanocrystals for enhancing radiosensitization effects.
Cancer theranostics combines therapeutic and diagnostic capabilities into a single system to treat cancer efficiently. Biocompatible nanomaterials can be engineered to exhibit cancer theranostic functions, for instance radiosensitization and photoluminescence. In this study, trivalent Bi and Eu ions were co-substituted into the lattice of hydroxyapatite (Bi(III):Eu(III) HAp) to develop a cancer theranostic nanocrystal. Bi provides radiosensitization capabilities while Eu imparts photoluminescence properties. To complement the radiotherapeutic function, l-buthionine sulfoximine (l-BSO) was adsorbed onto the nanocrystal surface. l-BSO inhibits the biosynthesis of cellular antioxidants, which can enhance radiosensitization effects. The Bi(III):Eu(III) HAp nanocrystals were prepared via a hydrothermal method. Structural and compositional analyses showed that both Bi and Eu ions were substituted into the HAp lattice. l-BSO was adsorbed onto the surface via electrostatic interactions between the charged carboxyl and amino groups of l-BSO and the surface ions of the nanocrystals. The adsorption followed the Langmuir isotherm model, implying a homogeneous monolayer adsorption. The l-BSO adsorbed Bi(III):Eu(III) HAp nanocrystals were found to have negligible cytotoxicity except the setting with l-BSO adsorbed amounts of 0.44 μmol/m2. This l-BSO amount was found to be high enough to elicit cytotoxicity due to l-BSO being released and causing excessive antioxidant depletion. Gamma ray irradiation clearly activated the cytotoxicity of the samples and increased the cell death rate, confirming radiosensitization abilities. At a constant amount of nanocrystals, the cell death rate increases with l-BSO concentration. This indicates that l-BSO can enhance the radiosensitization effect of the Bi(III):Eu(III) HAp nanocrystals.


Interpreting the pathogenicity of Joubert syndrome missense variants in Caenorhabditis elegans.
Ciliopathies are inherited disorders caused by defects in motile and non-motile (primary) cilia. Ciliopathy syndromes and associated gene variants are often highly pleiotropic and represent exemplars for interrogating genotype-phenotype correlations. Towards understanding disease mechanisms in the context of ciliopathy mutations, we have used a leading model organism for cilia and ciliopathy research, Caenorhabditis elegans, together with gene editing, to characterise two missense variants (P74S and G155S) in mksr-2/B9D2 associated with Joubert syndrome (JBTS). B9D2 functions within the Meckel syndrome (MKS) module at the ciliary base transition zone (TZ) compartment and regulates the molecular composition and sensory/signalling functions of the cilium. Quantitative assays of cilium/TZ structure and function, together with knock-in reporters, confirm that both variant alleles are pathogenic in worms. G155S causes a more severe overall phenotype and disrupts endogenous MKSR-2 organisation at the TZ. Recapitulation of the patient biallelic genotype shows that compound heterozygous worms phenocopy worms homozygous for P74S. The P74S and G155S alleles also reveal evidence of a very close functional association between the B9D2-associated B9 complex and MKS-2/TMEM216. Together, these data establish C. elegans as a model for interpreting JBTS mutations and provide further insight into MKS module organisation. This article has an associated First Person interview with the first author of the paper.


[Certain mechanisms of development of types of body tolerance to acute hypobaric hypoxia].
The regulatory mechanisms of individual rat resistance to acute hypobaric hypoxia were studied using the functional indices of the central nervous system (neurochemical and behavioral) and the hematopoietic system. The resistance to hypoxia was evaluated by the time of attitudinal reflex maintenance and recovery after decompression to a simulated altitude of 11200 m. Animals with different types of tolerance to hypoxia demonstrated different metabolic backgrounds of neurochemical processes (which were most balanced in moderately resistant rats). This agrees with the differences in active behavior and adaptive efficiency of these animals exposed to mild open-field stress. High functional activity of erythropoiesis and early leukocytic response were observed in hypoxia-tolerant rats.


Cladosporium sphaerospermum extract inhibits quorum sensing associated virulence factors of Serratia marcescens.
Serratia marcescens is now becoming a propensity for its highly antimicrobial-resistant clinical infections. Currently, it provides a novel strategy to prevent and control microbial infection by regulating S. marcescens quorum sensing (QS). Deep-sea-derived fungi are rich in QS bioactive constituents. In this work, the extracts from Cladosporium sphaerospermum SCSGAF0054 showed potent QS-related virulence factors and biofilm-inhibiting activities against S. marcescens NJ01. The swimming motility and multiple virulence factors such as prodigiosin, exopolysaccharide (EPS), lipase, protease and hemolysin were moderately inhibited by the extracts at varied concentrations. The confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM) images revealed that C. sphaerospermum extracts moderately arrested biofilm formation and cell viability. Further, real-time quantitative PCR (RT-qPCR) analysis revealed that expressions of genes associated with virulence factors, flhD, fimA, fimC, bsmA, bsmB, pigA, pigC, and shlA, were significantly down-regulated compared with control. In addition, the extracts combined with imipenem inhibited the QS system of S. marcescens NJ01, disrupted its preformed biofilm, released the intra-biofilm bacteria and killed the bacteria gradually. Therefore, the extracts combined with imipenem can partially restore bacterial drug sensitivity. These results suggest that the extracts from SCSGAF0054 effectively interfere with the QS system to treat S. marcescens infection alone or combining with classical antimicrobial drugs.


Efficacy and Safety of Primary Stereotactic Radiosurgery in Patients With Intraventricular Meningiomas.
Primary stereotactic radiosurgery for intraventricular meningiomas remains controversial owing to the potential for life-threatening peritumoral edema and lack of long-term follow-up data. We review the literature and present the largest series to assess efficacy and safety of primary stereotactic radiosurgery. A systematic review of the literature for primary stereotactic radiosurgery for intraventricular meningiomas was conducted. The retrospective series presented here comprised 33 patients who received primary stereotactic radiosurgery between 1999 and 2015 for a radiologically detected intraventricular meningioma. Demographic, diagnostic, and therapeutic data were extracted from medical records, imaging, and treatment-planning systems. Both standalone and pooled analysis were performed. The mean patient age was 53 years, and 24 patients (73%) were female. The median Karnofsky performance status pretreatment was 80 (range, 60-100). The majority of lesions were located in the lateral ventricles (n = 32; 97%). The mean tumor volume was 8.7 cm3 (range, 0.6-44.55 cm3). The mean delivered dose was 1390.9 cGy. Complete imaging follow-up data were available for 21 patients (64%). Of those, 14 (67%) showed partial or marginal response, 7 (33%) had stable disease, and no patient progressed per Response Assessment in Neuro-Oncology criteria. On last follow-up, 32 patients (97%) had significant improvement in performance status and a decrease in pretreatment symptoms. No high-grade Common Terminology Criteria for Adverse Events (version 5.0) toxicity was observed with the dose range employed. Primary stereotactic radiosurgery for intraventricular meningiomas shows excellent treatment efficacy and low toxicity in patients with a long follow-up period. The best therapeutic algorithm remains to be established leveraging further clinical investigation.


Incidence and distribution of coeliac disease in Campania (Italy): 2011-2013.
There exists a wide variation in the reported incidence of coeliac disease in recent decades. We aimed to evaluate the incidence rate of coeliac diagnoses performed in an Italian region, Campania, between 2011 and 2013 and its variation therein. All coeliac diagnoses made from 2011 to 2013 and registered within the Campania coeliac disease register (CeliacDB) were identified. Incidence rates were analysed by sex, age and province of residence, with a Poisson model fitted to determine incidence rate ratios. We found 2049 coeliac disease diagnoses registered in the CeliacDB between 2011 and 2013; 1441 of these patients were female (70.4%) and 1059 were aged less than 19 years (51.7%). The overall incidence of coeliac disease in Campania was 11.8 per 100,000 person-years (95% CI 11.3-12.3) during the study period, with marked variation by age [27.4 per 100,000 person-years (95% CI 25.8-29.1) in children under 19 years of age and 7.3 per 100,000 (95% CI 6.8-7.8) in adults] and sex [16.1 per 100,000 person-years in females (95% CI 15.3-16.9) and 7.2 per 100,000 person-years in males (95% CI 6.6-7.8)]. Coeliac disease incidence was roughly similar in Naples, Salerno, Caserta and Avellino, but about half in Benevento. More than 80% of our study population was diagnosed by the combination of positive antitransglutaminase IgA and Marsh 3. More than half of the patients were symptomatic at the time of coeliac disease diagnosis (39.7% had a classical presentation and 21.1% a non-classical one according to the Oslo definition). Coeliac disease incidence was roughly similar among Campania provinces, except in Benevento where it was about half, probably due to less awareness of coeliac disease in this area. The incidence of coeliac disease in Campania appears to be lower than that reported by most of the previous literature, suggesting the necessity of new coeliac awareness programmes.


An efficient two-step subcellular fractionation method for the enrichment of insulin granules from INS-1 cells.
Insulin is one of the key regulators for blood glucose homeostasis. More than 99% of insulin is secreted from the pancreatic β-cells. Within each β-cell, insulin is packaged and processed in insulin secretary granules (ISGs) before its exocytosis. Insulin secretion is a complicated but well-organized dynamic process that includes the budding of immature ISGs (iISGs) from the trans-Golgi network, iISG maturation, and mature ISG (mISG) fusion with plasma membrane. However, the molecular mechanisms involved in this process are largely unknown. It is therefore crucial to separate and enrich iISGs and mISGs before determining their distinct characteristics and protein contents. Here, we developed an efficient two-step subcellular fractionation method for the enrichment of iISGs and mISGs from INS-1 cells: OptiPrep gradient purification followed by Percoll solution purification. We demonstrated that by using this method, iISGs and mISGs can be successfully distinguished and enriched. This method can be easily adapted to investigate SGs in other cells or tissues, thereby providing a useful tool for elucidating the mechanisms of granule maturation and secretion.


The effect of competition on the relationship between the introduction of the DRG system and quality of care in Korea.
The diagnosis-related group-based prospective payment programme was introduced in Korea in 1997 as a pilot programme to control health spending. In July 2013, the programme was implemented throughout the nation. The aim of our study is to evaluate the relationship between quality of care and market competition following the introduction of the new payment system in Korea. We conduct an observational analysis using National Health Insurance claim data from 2011 to 2014. We analyse data on readmission within 30 days, length of stay, and number of outpatient visits for 1742 hospitals and 821 912 cases. We use a generalized estimating equation model to evaluate readmission within 30 days and number of outpatient visits and a multi-level regression model to assess length of stay. Total readmission within 30 days is 10 727 (1.3%). High competition areas present a lower risk of readmission [odds ratio (OR): 0.95, P: 0.0277], a longer length of stay (1%, P < 0.0001), and an increased number of outpatient visits (Relative Risk: 1.11, P: 0.0011) as compared with moderate competition areas. Risk of readmission is higher in low competition areas as compared with moderate competition areas (OR: 1.21, P < 0.0001). The effects of the introduction of the new payment system differed by degree of market competition. Thus, evaluation about the effect of new payment system on hospital performance should be measured in combination with the degree of hospital market structure.


Studies on the pre-alpha-lipoprotein of human serum. I. characterization of an albumin-containing lipoprotein.
In this study an albumin-containing lipoprotein is isolated by column affinity chromatography and electrophoresis. Its electrophoretic mobility is faster than that of albumin in agarose gel. Immunologically it reacts with anti-albumin and anti-apolipoprotein-A-I. No reaction is obtained against antibodies to any of the other previously known lipoportein families: Apo-AII, Apo-B, Lp(a), Apo-C, LpX, Apo-D or Apo-E ('the arginine rich peptide'). It differs from purified albumin in having higher amounts of glycine, serine and glutamic acid. Moreover, the molecular weight is estimated to 80,000 as compared to 67,000 for albumin.


Carbon source transport, nucleotide levels, and stable RNA synthesis in a mutant strain of Escherichia coli.
We have previously described a temperature-sensitive mutant of Escherichia coli, 2S142 (rel-, met-, rns-, ilv-, ts-) which shows specific inhibition of stable RNA synthesis at 42 degrees C. This mutation mimics a carbon source downshift in that the decay of guanosine 5'-diphosphate, 3'-diphosphate (ppGpp) is inhibited at the restrictive temperature. In this paper we show that the temperature-sensitive lesion in 2S142 does affect the uptake of glucose or alpha-D-methylglucopyranoside (alpha DMG) at 42 degrees C. However, restoration of glucose or alpha DMG uptake by the insertion of a constitutive galactose permease gene or further restriction of glucose uptake by insertion of a ptsG mutation into 2S142 have no effect on rRNA synthesis at 42 degrees C (although ppGpp levels are lowered in both cases). Furthermore, while restriction of uptake at 42 degrees C varies widely from carbon source to carbon source, severe restriction of rRNA synthesis is observed on all carbon sources tested at 42 degrees C. Levels of glycolytic intermediates, adenylate energy charge, ATP levels, and cAMP levels are all unaffected at the restrictive temperature. GTP levels decrease at 42 degrees C in glucose grown cells but that also does not appear to be related to the decrease in rRNA synthesis. These data were interpreted to suggest that the restriction of stable RNA synthesis in 2S142 at 42 degrees C can not be explained on the basis of decreased uptake and/or metabolism of carbon source. "Phantom spot" levels do decrease in 2S142 at 42 degrees C. In fact, "phantom spot" is the only putative regulatory molecule which correlates with restriction of rRNA synthesis on all carbon sources tested.


Comparing BamHI-W and CE-marked assays to detect circulating Epstein-Barr Virus (EBV) DNA of nasopharyngeal cancer patients in a non-endemic area.
Plasma Epstein-Barr Virus (EBV)-DNA is a well-established prognostic biomarker in nasopharyngeal carcinoma (NPC). Different methods for assessment include single-copy gene targeted, European Conformity (CE)-marked assays, which are mostly employed in non-endemic settings, vs multiple-copy gene targeted, in-house BamHI-W based assays, which currently represent the most widely used method for EBV-DNA quantification. To date, evidence concerning the commutability of these different assays is still limited. From August 2016 to March 2018, 124 plasma and 124 whole blood (WB) samples from 93 NPC patients were collected at different time-points for each patient. EBV-DNA viral load was quantified in pre- (n = 12) and post-treatment (n = 9), follow-up (n = 53), and recurrent/metastatic (R/M) (n = 50) phase. For each sample, one in-house BamHI-W vs three different CE-marked plasma assays were compared; the performance of plasma vs WB matrix was also assessed. Quantitative agreement of EBV-DNA values was evaluated by linear correlation and Bland-Altman analysis. A statistically significant (p = 0.0001) agreement between all CE-marked and the BamHI-W assays was found using plasma matrix, regardless of clinical phase. The results obtained in copies/ml were comparable to those expressed in IU/ml. When using WB matrix, the number of positive detections increased in the post-treatment phase. Our retrospective comparison supported an agreement between Plasma BamHI-W and CE-marked assays in measuring EBV-DNA for non-endemic NPC patients. There were no significant interferences from different measurement units (IU/ml vs copies/ml). Further evaluations are needed to better clarify the role of WB.


RNA 4 sequences from cucumber mosaic virus subgroups I and II.
The complete sequences of RNA 4 from cucumber mosaic virus (CMV) strains Ny (subgroup I) and Sn (subgroup II) have been determined and compared to all other known complete CMV RNA 4 sequences. The identification of a unique EcoRI site, present only in subgroup-II RNA 4 sequences, provides the basis for a simple method of classifying CMV isolates.


Plasticity of adult stem cells.
Recent years have seen much excitement over the possibility that adult mammalian stem cells may be capable of differentiating across tissue lineage boundaries, and as such may represent novel, accessible, and very versatile effectors of therapeutic tissue regeneration. Yet studies proposing such "plasticity" of adult somatic stem cells remain controversial, and in general, existing evidence suggests that in vivo such unexpected transformations are exceedingly rare and in some cases can be accounted for by equally unexpected alternative explanations.


Vascular masses of the head and neck.
The classification, clinical presentation, natural history and imaging appearances of vascular masses of the head and neck are reviewed. The radiological appearances of congenital vascular masses (infantile haemangiomas and other congenital vascular tumours, high-flow and low-flow vascular malformations), acquired vascular masses (benign and malignant tumours, non-neoplastic lesions) and other hypervascular masses are described and illustrated, together with consideration of image-guided interventions.


Human and bovine brain cathepsin L and cathepsin H: purification, physico-chemical properties, and specificity.
Cathespin L (EC 22.214.171.124) and cathepsin H (EC 188.8.131.52) have been purified from brain cortex to apparent homogeneity by a simultaneous procedure involving acid extraction of homogenate at pH 4.2, ammonium sulfate fractionation (30-80%), chromatography on pepstatin-Sepharose, CM-Sephadex C-50, DEAE-Sephadex A-50, phenyl- and concanavalin A-Sepharose and isoelectric focusing. Cathepsin L and cathepsin H were assayed in the presence of dithiothreitol and Na2EDTA (2 mM each) with Z-Phe-Arg-NHMec (pH 5.5) and Lys-NNa (pH 6.5) respectively. Cathepsin L consists of 2 polypeptide chains with Mr 25,000 and 5,000, Mr of cathepsin H is 28,000. Cathepsin L exists in brain tissue in two multiple forms with pI values 5.7 and 5.9, pI of cathepsin H is 6.8. Substrate specificity of these thiol proteinases was tested with proteins (pyridoxyl-hemoglobin, azocasein) and low Mr naphthylamide and methylcoumarylamide substrates: Lys-NNa, Arg-NNa, Dz-Arg-NNa, Z-Arg-Arg-NNaOMe, Z-Phe-Arg-NHMec, Z-Phe, Val-Arg-NHMec, Z-Gly-Gly-Arg-NHMec. Z-Phe-Arg-NHMec is the best substrate for cathepsin L (KM = 5 microM, Kcat = 21 s-1), Arg-NNa--for cathepsin H (KM = 0.1 mM, Kcat = 1.93 s-1), being endoaminopeptidase cathepsin H also hydrolyses Bz-Arg-NNa (KM = 0.7 mM, Kcat = 1.3 s-1). Both proteinases are inhibited by traditional inhibitors of cysteine proteinases and E-64, but leupeptin turned to be more effective inhibitor of cathepsin L (Ki = 2.4 nM) than of cathepsin H (Ki = 9.2 microM), the latter enzyme being sensitive to puromycin and benzethonium chloride as well.(ABSTRACT TRUNCATED AT 250 WORDS)


Voluntary physical activity protects from susceptibility to skeletal muscle contraction-induced injury but worsens heart function in mdx mice.
It is well known that inactivity/activity influences skeletal muscle physiological characteristics. However, the effects of inactivity/activity on muscle weakness and increased susceptibility to muscle contraction-induced injury have not been extensively studied in mdx mice, a murine model of Duchenne muscular dystrophy with dystrophin deficiency. In the present study, we demonstrate that inactivity (ie, leg immobilization) worsened the muscle weakness and the susceptibility to contraction-induced injury in mdx mice. Inactivity also mimicked these two dystrophic features in wild-type mice. In contrast, we demonstrate that these parameters can be improved by activity (ie, voluntary wheel running) in mdx mice. Biochemical analyses indicate that the changes induced by inactivity/activity were not related to fiber-type transition but were associated with altered expression of different genes involved in fiber growth (GDF8), structure (Actg1), and calcium homeostasis (Stim1 and Jph1). However, activity reduced left ventricular function (ie, ejection and shortening fractions) in mdx, but not C57, mice. Altogether, our study suggests that muscle weakness and susceptibility to contraction-induced injury in dystrophic muscle could be attributable, at least in part, to inactivity. It also suggests that activity exerts a beneficial effect on dystrophic skeletal muscle but not on the heart.


Medicaid insurance as primary payer predicts increased mortality after total hip replacement in the state inpatient databases of California, Florida and New York.
To confirm the relationship between primary payer status as a predictor of increased perioperative risks and post-operative outcomes after total hip replacements. Retrospective cohort study. Administrative database study using 2007-2011 data from California, Florida, and New York from the State Inpatient Databases (SID), Healthcare Cost and Utilization Project, Agency for Healthcare Research and Quality. 295,572 patients age≥18years old who underwent total hip replacement with non-missing insurance data were collected, using International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) diagnoses and procedures code (ICD-9-CM code 81.51). Patients underwent total hip replacement. Patients were cohorted by insurance type as either Medicare, Medicaid, Uninsured, Other, and Private Insurance. Demographic characteristics and comorbidities were compared. Unadjusted rates of in-hospital mortality, postoperative complications, LOS, 30-day, and 90-day readmission status were compared. Adjusted odds ratios were calculated for our outcomes using multivariate linear and logistic regression models fitted to our data. Medicaid patients incurred a 125% increase in the odds of in-hospital mortality compared to those with Private Insurance (OR 2.25, 99% CI 1.01-5.01). Medicaid payer status was associated with the highest statistically significant adjusted odds of mortality, any complication (OR, 1.26), cardiovascular complications (OR, 1.37), and infectious complications (OR, 1.66) when compared with Private Insurance. Medicaid patients had the highest statistically significant adjusted odds of 30-day (OR, 1.63) and 90-day readmission (OR, 1.58) and the longest adjusted LOS. We found higher unadjusted rates and risk adjusted odds ratios of postoperative mortality, morbidity, LOS, and readmissions for patients with Medicaid insurance as compared to patients with Private Insurance. Our study shows that primary payer status serves as a predictor of perioperative risks and that primary payer status should be viewed as a peri-operative risk factor.


Topical Review: Crowdsourcing as a Novel Approach to Qualitative Research.
To describe a novel, five-phase approach to collecting qualitative data from hard-to-reach populations using crowdsourcing methods. Drawing from experiences across recent studies with type 1 diabetes and congenital heart disease stakeholders, we describe five phases of crowdsourcing methodology, an innovative approach to conducting qualitative research within an online environment, and discuss relevant practical and ethical issues. Phases of crowdsourcing methodology are: (I) Preparing; (II) Forming Crowds; (III) Collecting Crowdsourced Data; (IV) Coding and Analyzing Crowdsourced Data; and (V) Generating and Disseminating Findings. Iterative feedback from stakeholders is obtained in all five phases. Practical and ethical issues include accessing diverse stakeholders, emotional engagement of crowd participants, responsiveness and transparency of crowdsourcing methodology, and limited personal contact with crowd participants. Crowdsourcing is an innovative, efficient, feasible, and timely approach to engaging hard-to-reach populations in qualitative research.


Complete protection by a single-dose skin patch-delivered SARS-CoV-2 spike vaccine.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected more than 160 million people and resulted in more than 3.3 million deaths, and despite the availability of multiple vaccines, the world still faces many challenges with their rollout. Here, we use the high-density microarray patch (HD-MAP) to deliver a SARS-CoV-2 spike subunit vaccine directly to the skin. We show that the vaccine is thermostable on the patches, with patch delivery enhancing both cellular and antibody immune responses. Elicited antibodies potently neutralize clinically relevant isolates including the Alpha and Beta variants. Last, a single dose of HD-MAP–delivered spike provided complete protection from a lethal virus challenge in an ACE2-transgenic mouse model. Collectively, these data show that HD-MAP delivery of a SARS-CoV-2 vaccine was superior to traditional needle-and-syringe vaccination and may be a significant addition to the ongoing COVID-19 (coronavirus disease 2019) pandemic.


Radiofrequency ablation with a new perfused-cooled electrode using a single pump: an experimental study in ex vivo bovine liver.
The purpose of this study was to assess the efficacy of a new perfused-cooled electrode that uses a single pump for creating a large ablation zone in explanted bovine liver. This was done by comparing with the radiofrequency (RF) ablation zones that were created with a monopolar cooled electrode to the RF ablation zones that were created by the new perfused-cooled electrode. We developed a new perfused-cooled electrode that uses a single pump by modifying a 17-gauge cooled electrode (Radionics) with a 2.5-cm outer metallic sheath (15-gauge) in order to allow use of the internal cooling water (5.85 % hypertonic saline) for the infused saline. Thirty ablation zones were created in explanted bovine livers (12-min ablation cycle; pulsed technique; 2000 mA, maximum) with three different regimens: group A, RF ablation with the 17-gauge cooled electrode; group B, RF ablation with the 15-gauge cooled electrode; group C, RF ablation with the perfused-cooled electrode. T2-weighted magnetic resonance (MR) imaging was obtained immediately after RF ablation for calculating volumes of the ablation zone. Following MR imaging, the ablation zones were excised and measured for transverse diameters and vertical diameters. The transverse diameter, vertical diameter, and the calculated volumes of the ablation zones on MR imaging were compared among the groups. Ablation zones created with the perfused-cooled electrode (group C) were significantly larger than those created with the 17-gauge cooled electrode (group A) and the 15-gauge cooled electrode (group B) according to the transverse diameter and vertical diameter on the gross specimens (p < 0.05): 3.6 +/- 0.38 cm and 4.4 +/- 0.20 cm in group A, 3.7 +/- 0.08 cm and 4.6 +/- 0.16 cm in group B, and 5.4 +/- 0.65 cm and 6.0 +/- 0.56 cm in group C, respectively. On the MR imaging, the calculated volumes of the ablation zones in group C were significantly larger than those in groups A and B (p < 0.05): 23.1 +/- 8.7 cm3 in group A, 28.9 +/- 5.7 cm3 in group B, and 80.0 +/- 34 cm3 in group C, respectively. A new perfused-cooled electrode using a single pump could efficiently increase the size of the ablation zone in liver compared with a monopolar cooled electrode, and this was due to its simultaneous use of internal cooling and saline infusion.


Staying in touch: the molecular era of organelle contact sites.
Membrane contact sites (MCS) are close appositions between two organelles that facilitate both signaling and the passage of ions and lipids from one cellular compartment to another. Despite the fact that MCS have been observed for over 50 years now, we still know very little about the molecular machinery required to create them or their structure, function and regulation. In this review, we focus on the three best-characterized contact sites to date: the nucleus-vacuole junction and mitochondria-ER and plasma membrane-ER contact sites. In addition, we discuss principles arising from recent research and highlight several unanswered questions in the field.


Efficacy and safety of extended release oxybutynin for the treatment of urge incontinence: an analysis of data from 3 flexible dosing studies.
We assessed the efficacy and safety of extended release (ER) oxybutynin at individualized doses for urge urinary incontinence. Data were combined from 3 flexible dosing studies of a total of 420 patients with urge urinary incontinence or mixed incontinence in whom ER oxybutynin dose adjustments were made to achieve the optimal balance between efficacy and tolerability, as judged by each participant. Individual doses were adjusted in 5 mg increments across a dose range of 5 to 30 mg daily. Efficacy was assessed during maintenance therapy using a 7-day diary. Data on 368 patients who completed dose adjustment were included in the analysis. In 47% of patients the preferred dose was greater than 10 mg daily. Individualized doses of ER oxybutynin produced a 79.3% decrease from baseline in the total of all incontinence episodes and an 83.2% decrease in urge episodes. Of patients 81% achieved a decrease of at least 70% in all incontinence episodes and 43% achieved total dryness. Moderate or severe dry mouth was reported by 23% of patients and it was cited as the cause for early withdrawal by 1.4% who completed dose adjustment. Therapy was well tolerated. In the pooled sample overall 6.7% of patients cited adverse events as the reason for discontinuing. Individualized dosing with ER oxybutynin was associated with highly effective and well tolerated control of urinary incontinence with more than 40% of patients achieving total dryness.


Repeat operations in patients with anal fistula, a retrospective study across England and Wales.
This article reports the frequency of repeat operations including waiting times within the National Health Service (NHS) of England and Wales. Retrospective study on repeat operations for anal fistula (AF) performed between 1st January 2010 and 31st December 2016. Data were extracted from the national registry of data entered into Hospital Episode Statistics (HES). Patient factors (age, sex, self-declared ethnicity) and geographical location were tested for association with repeat operations and time to the second operation. We analysed 36,223 patients that had an operation for AF within 148 NHS trusts. The median follow-up time was 28 months. The majority of patients (67.4%) had only one operation. Eighty-five per cent of them remained under the care of a single consultant. Six per cent of the repeat surgeries occurred in at least three different treatment sites. Young age and female sex were associated with higher rates of repeat operations. Non-declared and Black or Black British ethnicity were associated with fewer operations. The median waiting time between the first and second operations was 27.4 weeks (IQR: 14.7-55.3); between the second and third 28.0 weeks (IQR: 14.7-57.0); between the third and fourth 29.0 weeks. This large real world population-based study shows that the majority of patients with AF undergo only one operation. Patients requiring multiple procedures tend to stay under the care of a small number of consultants but waiting times between operations are long. There is a geographical variation in the number of operations and the time between them.


Synthesis of the indene, THF, and pyrrolidine skeletons by Lewis acid mediated cycloaddition of methylenecyclopropanes with aldehydes, N-tosyl aldimines, and acetals.
Methylenecyclopropanes (MCPs 1) react with aldehydes, N-tosyl aldimines, and acetals to give the corresponding indene, THF, and pyrrolidine cycloaddition products in the presence of BF3 x OEt2 under mild reaction conditions. Some special transformations of MCPs 1 with aldehydes have been reported in this paper. A plausible reaction mechanism has been discussed, which is based on a deuterium-labeling experiment and the Prins-type reaction mechanism.


Quantitative autoradiographic study on tyrosine hydroxylase mRNA with in situ hybridization and alpha 2 adrenergic receptor binding in the locus coeruleus of the spontaneously hypertensive rat.
alpha-2 Adrenergic (A2) receptors and tyrosine hydroxylase (TH) mRNA in the locus coeruleus (LC) were studied using [125I]iodoclonidine and [35S]TH oligonucleotide probe. Spontaneously hypertensive (SHR) rats contained less TH mRNA at their prehypertensive, but not at the well-established hypertensive stage, than age-matched Wistar-Kyoto rats. Furthermore, there is an up-regulation of A2 receptors in SHR rats which is parallel to their blood pressure elevation. The present data suggest that increased A2 receptors in conjunction with TH mRNA reduction in the LC are associated with initiation, but not maintenance of genetic hypertension.


New non-hydroxamic ADAMTS-5 inhibitors based on the 1,2,4-triazole-3-thiol scaffold.
In this Letter we describe the design, synthesis, screening, and optimization of a new family of ADAMTS-5 inhibitors. These inhibitors display an original 1,2,4-triazole-3-thiol scaffold as a putative zinc binding-group. In vitro results are rationalized by in silico docking of the compounds in ADAMTS-5's crystal structure.


Mapping the Patient-Reported Outcomes Measurement Information System (PROMIS-29) to EQ-5D-5L.
The Patient-Reported Outcomes Measurement Information System (PROMIS-29) is gaining popularity as healthcare system funders increasingly seek value-based care. However, it is limited in its ability to estimate utilities and thus inform economic evaluations. This study develops the first mapping algorithm for estimating EuroQol 5-Dimension 5-Level (EQ-5D-5L) utilities from PROMIS-29 responses using a large dataset and through extensive comparisons between econometric models. An online survey was conducted to collect responses to PROMIS-29 and EQ-5D-5L from the general Australian population (N = 3013). Direct and indirect mapping methods were explored, including linear regression, Tobit, generalised linear model, censored regression model, beta regression (Betamix), the adjusted limited dependent variable mixture model (ALDVMM) and generalised ordered logit. The most robust model was selected by assessing the performance based on average ten-fold cross-validation geometric mean absolute error and geometric mean squared error, the predicted mean, maximum and minimum utilities, as well as the fitting across the entire distribution. The direct approach using ALDVMM was considered the preferred model based on lowest geometric mean absolute error and geometric mean squared error in cross-validation (0.0882, 0.0299) and its superiority in predicting the actual observed mean, full health states and lower utility extremes. The robustness and precision in prediction across the entire distribution of utilities with ALDVMM suggest it is an accurate and valid mapping algorithm. Moreover, the suggested mapping algorithm outperformed previously published algorithms using Australian data, indicating the validity of this model for economic evaluations. This study developed a robust algorithm to estimate EQ-5D-5L utilities from PROMIS-29. Consistent with the recent literature, the ALDVMM outperformed all other econometric models considered in this study, suggesting that the mixture models have relatively better performance and are an ideal candidate model for mapping.


Hypertrophic Cardiomyopathy in Pregnancy.
Hypertrophic cardiomyopathy (HCM) is the most common genetic cardiac condition and highly heterogeneous. Echocardiography and genetic and clinical screening have led to detection in women of childbearing age. Maternal and fetal outcomes among women with HCM are favorable. Genetic counseling is recommended. Prepregnancy clinical evaluation and risk assessment are paramount in ensuring optimal outcomes. Most women carry moderate risk of morbidity, have clinical evaluations and echocardiography each trimester, and deliver vaginally. Those who are symptomatic or have significant left ventricular outflow obstruction or recurrent arrhythmias prior to pregnancy are at higher risk and should be monitored at least monthly.


Two new species of the pseudoscorpion subfamily Lamprochernetinae Beier, 1932 from Guizhou, China (Pseudoscorpiones: Chernetidae).
Two new species of the pseudoscorpion subfamily Lamprochernetinae Beier, 1932 are described from Guizhou Province, China, Nudochernes pseudotroglobius sp. nov. and Megachernes biyunensis sp. nov., with detailed diagnoses, descriptions and illustrations.


Baseline characteristics, management practices, and in-hospital outcomes of patients with acute coronary syndromes: Results of the Saudi project for assessment of coronary events (SPACE) registry.
The Saudi Project for Assessment of Coronary Events (SPACE) registry is the first in Saudi Arabia to study the clinical features, management, and in-hospital outcomes of acute coronary syndrome (ACS) patients. We conducted a prospective registry study in 17 hospitals in Saudi Arabia between December 2005 and December 2007. ACS patients included those with ST-elevation myocardial infarction (STEMI), non-ST elevation myocardial infarction and unstable angina; both were reported collectively as NSTEACS (non-ST elevation acute coronary syndrome). 5055 patients were enrolled with mean age ± SD of 58 ± 12.9 years; 77.4% men, 82.4% Saudi nationals; 41.5% had STEMI, and 5.1% arrived at the hospital by ambulance. History of diabetes mellitus was present in 58.1%, hypertension in 55.3%, hyperlipidemia in 41.1%, and 32.8% were current smokers; all these were more common in NSTEACS patients, except for smoking (all P < 0.0001). In-hospital medications were: aspirin (97.7%), clopidogrel (83.7%), beta-blockers (81.6%), angiotensin converting enzyme inhibitors/angiotensin receptor blockers (75.1%), and statins (93.3%). Median time from symptom onset to hospital arrival for STEMI patients was 150 min (IQR: 223), 17.5% had primary percutaneous coronary intervention (PCI), 69.1% had thrombolytic therapy, and 14.8% received it at less than 30 min of hospital arrival. In-hospital outcomes included recurrent myocardial infarction (1.5%), recurrent ischemia (12.6%), cardiogenic shock (4.3%), stroke (0.9%), major bleeding (1.3%). In-hospital mortality was 3.0%. ACS patients in Saudi Arabia present at a younger age, have much higher prevalence of diabetes mellitus, less access to ambulance use, delayed treatment by thrombolytic therapy, and less primary PCI compared with patients in the developed countries. This is the first national ACS registry in our country and it demonstrated knowledge-care gaps that require further improvements.


Uterine balloon therapy to treat menorrhagia.
To study the clinical efficacy, safety and acceptability of the thermal balloon endometrial ablation (TBEA) in patients with dysfunctional uterine bleeding. Thirteen patients with DUB who did not respond to medical treatment and dilatation and curettage consented to a trial of TBEA with EASY model of balloon catheter by Gynecare. All procedures were done under intravenous (i.v.) sedation and paracervical block. Patients were mostly discharged within 48 h. Follow-up of 2-19 months is reported. Transvaginal ultrasound and hysteroscopy were performed in six patients after 6 months of TBEA. Twelve patients (92.3%) reported a significant reduction in bleeding. Two patients (15.4%) experienced amenorrhea. Only two patients (15.4%) underwent subsequent hysterectomy, one for persistent menorrhagia and the other for severe pelvic pain. In successful patients of TBEA, transvaginal ultrasonography revealed marked reduction in endometrial thickness and no endometrium was visible in one patient who had developed amenorrhea. In successful cases follow-up hysteroscopy revealed scarring in more than two-thirds of the endometrium. Thermal balloon endometrial ablation is a safe, simple, effective, easy and minimally invasive procedure which can be done under i.v. sedation and paracervical block in an office setting. It has several advantages over hysterectomy, including preservation of the uterus, avoidance of surgical incision and potential to perform on an outpatient basis. It is a good alternative to hysteroscopic endometrial ablation with a comparable success rate with minimal risk and no limiting factors except the cost of the balloons. However, large scale randomized controlled studies are needed with TBEA and other endometrial ablation procedures.


The Logic of Transgenerational Inheritance: Timescales of Adaptation.
Myriad mechanisms have evolved to adapt to changing environments. Environmental stimuli alter organisms' physiology to create memories of previous environments. Whether these environmental memories can cross the generational barrier has interested scientists for centuries. The logic of passing on information from generation to generation is not well understood. When is it useful to remember ancestral conditions, and when might it be deleterious to continue to respond to a context that may no longer exist? The key might be found in understanding the environmental conditions that trigger long-lasting adaptive responses. We discuss the logic that biological systems may use to remember environmental conditions. Responses spanning different generational timescales employ different molecular machineries and may result from differences in the duration or intensity of the exposure. Understanding the molecular components of multigenerational inheritance and the logic underlying beneficial and maladaptive adaptations is fundamental to understanding how organisms acquire and transmit environmental memories across generations.


The molecular origins of multicellular transitions.
Multicellularity has evolved multiple times independently from a variety of ancestral unicellular lineages. Past research on multicellularity was focused more on explaining why it was repeatedly invented and less so on the molecular foundations associated with each transition. Several recent comparative functional analyses of microbial unicellular and multicellular genomes have begun to throw considerable light on the molecular commonalities exhibited by independent multicellular transitions. These have enabled the delineation of the likely functional components of the genetic toolkit required for multicellular existence and to surprising discoveries, such as the presence of several toolkit components in unicellular lineages. The study of these toolkit proteins in a unicellular context has begun yielding insights into their ancestral functions and how they were coopted for multicellular development.


On estimating energetic values of prey: implications in optimal diet models.
The accurate estimation of the amount of energy contained within a food item which is available to a predator is essential in tests of optimal foraging theories. Many studies of optimal foraging measure gross energy content of prey directly by bomb calorimetry. I suggest that a more realistic and accurate estimate of true "prey value" is available by calculating energy associated with the organic constituents of prey, and then subtracting away energy associated with insoluble and indigestible components. This methodology allows for a much more precise estimate of prey value (useable energy) and therefore a more realistic test of optimal foraging models.


Complement component C9-dependent cytosolic free Ca2+ rise and recovery in neutrophils.
The cytosolic free Ca2+ concentration in rat neutrophils was determined by ratiometric fluorometry and imaging of Fura-2. Transient elevations of cytosolic free Ca2+ concentration were provoked by addition of C9 to neutrophils pre-coated with C5b-8. The rate of rise of the cytosolic free Ca2+ concentration was dependent upon the concentration of C9. These changes in cytosolic free Ca2+ concentration occurred in the absence of cell lysis, since there was no release of Fura-2, and were the result of increased permeability to extracellular Ca2+. More than 96% of the rise in cytosolic free Ca2+ generation by C9 was dependent upon the presence of extracellular Ca2+, but did not occur via channels which were inhibited by the Ca2+ channel blocker SKF96365. The decrease in the permeability of the membrane to Ca2+ after C9 was not triggered by the rise in cytosolic free Ca2+. After attack by C9, individual neutrophils remained responsive to f-met-leu-phe, or further attack by C9, both producing Ca2+ transients. The recovery of the Ca2+ signal was consistent with the complement membrane attack complex generating a series of permeability thresholds in the plasma membrane. These data have implications for the understanding of the mechanisms underlying the inappropriate responsiveness of neutrophils at inflammatory sites.


Gain control of synaptic response function in cerebellar nuclear neurons by a calcium-activated potassium conductance.
Small conductance Ca(2+)-activated potassium (SK) current provides an important modulator of excitatory synaptic transmission, which undergoes plastic regulation via multiple mechanisms. We examined whether inhibitory input processing is also dependent on SK current in the cerebellar nuclei (CN) where inhibition provides the only route of information transfer from the cerebellar cortical Purkinje cells. We employed dynamic clamping in conjunction with computer simulations to address this question. We found that SK current plays a critical role in the inhibitory synaptic control of spiking output. Specifically, regulation of SK current density resulted in a gain control of spiking output, such that low SK current promoted large output signaling for large inhibitory cell input fluctuations due to Purkinje cell synchronization. In contrast, smaller nonsynchronized Purkinje cell input fluctuations were not amplified. Regulation of SK density in the CN therefore would likely lead to important consequences for the transmission of synchronized Purkinje cell activity to the motor system.


Novel C-xylosylflavones from the leaves and flowers of Iris gracilipes.
Two new C-glycosylflavones, apigenin 7,4'-dimethyl ether 6-C-β-[(4"'-acetyl-L-rhamnopyranosyl)-(1-->2)-xylopyranoside] (1) and apigenin 7,4'-dimethyl ether 6-C-β-L-rhamnopyranosyl-(1-->2)-xylopyranoside (2) were isolated from the leaves of Iris gracilipes (Iridaceae), along with two known flavonoids, swertiajaponin (3) and swertisin (4). C-Xylosylflavones 1 and 2 were elucidated by UV and NMR spectroscopy, mass spectrometry, and acid and alkaline hydrolyses. These novel compounds were also presented in the flowers.


Functional and structural alterations in different durations of untreated illness in the frontal and parietal lobe in major depressive disorder.
Major depressive disorder (MDD) is one of the most disabling illnesses that profoundly restricts psychosocial functions and impairs quality of life. However, the treatment rate of MDD is surprisingly low because the availability and acceptability of appropriate treatments are limited. Therefore, identifying whether and how treatment delay affects the brain and the initial time point of the alterations is imperative, but these changes have not been thoroughly explored. We investigated the functional and structural alterations of MDD for different durations of untreated illness (DUI) using regional homogeneity (ReHo) and voxel-based morphometry (VBM) with a sample of 125 treatment-naïve MDD patients and 100 healthy controls (HCs). The MDD patients were subgrouped based on the DUI, namely, DUI ≤ 1 M, 1 < DUI ≤ 6 M, 6 < DUI ≤ 12 M, and 12 < DUI ≤ 48 M. Subgroup comparison (MDD with different DUIs) was applied to compare ReHo and grey matter volume (GMV) extracted from clusters of regions with significant differences (the pooled MDD patients relative to HCs). Correlations and mediation effects were analysed to estimate the relationships between the functional and structural neuroimaging changes and clinical characteristics. MDD patients exhibited decreased ReHo in the left postcentral gyrus and precentral gyrus and reduced GMV in the left middle frontal gyrus and superior frontal gyrus relative to HCs. The initial functional abnormalities were detected after being untreated for 1 month, whereas this duration was 3 months for GMV reduction. Nevertheless, a transient increase in ReHo was observed after being untreated for 3 months. No significant differences were discovered between HCs and MDD patients with a DUI less than 1 month or among MDD patients with different DUIs in either ReHo or GMV. Longer DUI was related to reduced ReHo with GMV as mediator in MDD patients. We identified disassociated functional and anatomical alterations in treatment-naïve MDD patients at different time points in distinct brain regions at the early stage of the disease. Additionally, we also discovered that GMV mediated the relationship between a longer DUI and diminished ReHo in MDD patients, disclosing the latent deleterious and neuro-progressive implications of DUI on both the structure and function of the brain and indicating the necessity of early treatment of MDD.


Correction to: RNA sequencing identifies novel non-coding RNA and exon-specific effects associated with cigarette smoking.
Following publication of the original article [1], the authors provided an updated accession number in the "Availability of data and materials" section of the declarations.


Serum 1,25-dihydroxy vitamin D is inversely associated with body mass index.
Based on in vitro studies, it has been hypothesized that 1,25-dihydroxy vitamin D (1,25-vit D) may promote weight gain in humans, but previous studies have demonstrated conflicting results regarding the association between serum 1,25-vit D and body mass index (BMI). To evaluate the relation between serum 1,25-vit D and BMI. Two thousand one hundred and eighty-seven subjects, recruited from a metabolic and medical lifestyle management clinic, were included in a cross-sectional study. BMI, 25-hydroxy vitamin D (25-OH-vit D) and 1,25-vit D were measured. The cohort was divided according to BMI in five groups (<25, 25-29.9, 30-34.9, 35-39.9 and >39.9 kg/m(2)). Statistical analyses were performed with multiple linear regression models. Age and gender were used as explanatory covariates. With increasing BMI group, there was a significant decrease in both serum 25-OH-vit D and 1,25-vit D (P<0.001). Those with BMI >39.9 kg/m(2) had 24% lower serum 25-OH-vit D levels and 18% lower 1,25-vit D levels than those with BMI <25 kg/m(2). There is an inverse association between BMI and the serum levels of 25-OH-vit D and 1,25-vit D. This makes it highly unlikely that high levels of circulating 1,25-vit D contribute to the development of obesity.


First record of Gongylonema nepalensis in domestic and wild ruminants in Europe.
The gullet worm, Gongylonema pulchrum, is cosmopolitan in distribution, infecting a variety of mammals including domestic and wild ruminants. Gongylonema nepalensis recently collected from the esophageal epithelium of water buffaloes in Nepal was separated from G. pulchrum based on its distinctly shorter left spicule relative to body length and unique nucleotide sequences of the ribosomal RNA gene (rDNA) and the mitochondrial cytochrome c oxidase subunit 1 gene (cox-1). During meat inspections at four abattoirs on Sardinia Island, Italy, 25 Gongylonema worms were collected from one each individual of cattle (n=8), sheep (n=7), goats (n=4), and mouflon (Ovis aries musimon; n=6), and characterized morphologically and genetically. Intriguingly, all of the collected worms from these ruminants were G. nepalensis, exhibiting comparable body lengths to G. pulchrum in cattle from other regions but with significantly shorter left spicules like G. nepalensis (less than 20.9% of the entire body length in contrast to 21.8-65.6%, the reported proportion of G. pulchrum). Furthermore, the rDNA nucleotide sequences of these worms from different ruminant species on Sardinia Island were almost identical to each other and to Nepalese G. nepalensis isolates. With the exception of one worm from a sheep (displaying a single nucleotide substitution), the 369-bp cox-1 nucleotide sequences of all the Sardinian G. nepalensis isolates from the different host sources were absolutely identical, forming a clade with Nepalese G. nepalensis isolates and not G. pulchrum isolates. The present study reveals that G. nepalensis is not a local parasite in the Indian subcontinent (Nepal), but instead has a certain geographical distribution in Europe and takes several ruminant species as a definitive host.


Different right ventricular dysfunction and pulmonary coupling in acute heart failure according to the left ventricular ejection fraction.
Right ventricular (RV) dysfunction and pulmonary uncoupling are two acknowledged features associated with poor outcome, however few data defined RV adaptation across the different left ventricular ejection fraction (EF) cut-off. Additionally, less data are reported in patients with acute heart failure (AHF). The aim of present study was to analyse RV function in AHF patients presenting with either reduced or preserved EF. This is a multi-center observational study including 380 patients affected by AHF: 235 had AHF with reduced EF (AHFrEF) and 145 had AHF with preserved EF (AHFpEF). Pulmonary artery systolic pressure (PASP), tricuspid annular plane systolic excursion (TAPSE), S' wave velocity, and the RV end-diastolic diameter (RVEDD) were measured by echocardiography. TAPSE/PASP and S'/PASP ratios were calculated as non-invasive surrogates of RV-pulmonary arterial coupling. Factors associated with poor outcome were higher values of PASP (45 [40-55] mmHg vs 40 [35-46] mmHg; p < 0.001), RVEDD (44 [38-47] mm vs 37 [35-42] mm; p < 0.001), lower TAPSE values (17 [15-20] mm vs 20 [18-22] mm; p < 0.001) and S' wave (10 [8-12] cm/s vs 11 [10-13] cm/s; p < 0.001), reduced TAPSE/PASP (0.37 [0.29-0.47] vs 0.50 [0.40-0.60]; p < 0.001) and S'/PASP ratios (0.22 [0.18-0.28] vs 0.28 [0.22-0.34]; p < 0.001). However, the prognostic parameters differed according to the LVEF value: in AHFpEF S'/PASP between 0.22 and 0.29 and > 0.29 demonstrated a protective prognostic value (Respectively HR 0.29 (0.16-0.53), p < 0.001 and HR 0.22 [0.12-0.42], p < 0.001). Conversely, in AHFrEF, TAPSE <16 mm (HR 2.59 [1.67-4.03], p < 0.001), ICV > 21 mm (HR 1.17 [1.17-1.28], p = 0.001) and TAPSE/PASP <0.49 HR 1.92 [1.10-3.37], p = 0.023) were related to adverse outcome. RV adaptation and RV pulmonary coupling differ in AHF according to the level of LVEF. S' wave, and S'/PASP are associated with adverse outcome in patients with preserved EF; reduced TAPSE and TAPSE/PASP are better prognostic predictors in patients with reduced EF.


Body size and microclimate use in Neotropical granivorous ants.
The stability of tropical microclimates has left microclimate use by tropical species little unexplored. At La Selva Costa Rica, I related foraging activity at seed baits to humidity in two forests types. I recorded 38 and 35 ant species at seed baits in closed and open canopy forest. The microclimate 5 cm above the forest floor in the younger, Open Forest was warmer, drier, more variable, and more sensitive to current weather than in the older Closed Forest. Ant species within both forests foraged at different Vapor Pressure Deficits (kPa), a measure of the drying power of the air. VPD use was not confounded with diel activity patterns. Body size explained 46% of the variance in mean VPD use among ant species. Small ant species tended to forage in moist microclimates; large species tended to be microclimate generalists. Larger species were also more active in the drier Open Forest. Foraging activity by these assemblages varies 4-fold, and peaks close to the mean VPD for each habitat. The behavior of these assemblages suggest that 1) small ant species at La Selva potentially compete with the entire range of ant body sizes, whereas large ants forage when and where small ants are inactive; and 2) seeds dispersed to the forest floor at dawn will be consumed or further dispersed by a larger suite of ants species than those falling in the heat of the tropical afternoon.


Neurodevelopmental, emotional, and behavioural problems in Duchenne muscular dystrophy in relation to underlying dystrophin gene mutations.
Duchenne muscular dystrophy (DMD) is associated with neuropsychiatric disorders. The aim of the study was to characterize the DMD neuropsychiatric profile fully and to explore underlying genotype/phenotype associations. One hundred and thirty males with DMD (mean age 9y 10mo, range 5-17y) in four European centres were included and completed IQ assessment and a neurodevelopmental-screening questionnaire. Of these, 87 underwent comprehensive neuropsychiatric assessment using structured diagnostic interview and parent-reported questionnaires. The overall mean score on the neurodevelopmental questionnaire was significantly abnormal compared with the general population of children (p<0.001). On average, intelligence was below the population mean, with intellectual disability observed in 34 males (26%). Autistic spectrum disorder was identified in 18 (21%), hyperactivity in 21 (24%), and inattention in 38 (44%). Clinical levels of internalizing and externalizing problems were observed in 21 (24%) and 13 (15%) respectively. Over a third of males scored more than two measures of emotional, behavioural, or neurodevelopmental problems. Males with mutations at the 3' end of the DMD gene affecting all protein isoforms had higher rates of intellectual disability and clusters of symptoms. Males with DMD are at very high risk of neuropsychiatric disturbance, and this risk appears to increase with mutations at the 3' end of the gene. Patterns of symptom clusters suggest a DMD neuropsychiatric syndrome, which may require prompt evaluation and early intervention.


Cargo receptor Surf4 regulates endoplasmic reticulum export of proinsulin in pancreatic β-cells.
Insulin is an essential peptide hormone that maintains blood glucose levels. Although the mechanisms underlying insulin exocytosis have been investigated, the mechanism of proinsulin export from the endoplasmic reticulum (ER) remains unclear. Here, we demonstrated that Surf4, a cargo receptor homolog, regulates the ER export of proinsulin via its recruitment to ER exit sites (ERES). Under high-glucose conditions, Surf4 expression was upregulated, and Surf4 proteins mainly localized to the ER at a steady state and accumulated in the ERES, along with proinsulin in rat insulinoma INS-1 cells. Surf4-knockdown resulted in proinsulin retention in the ER and decreased the levels of mature insulin in secretory granules, thereby significantly reducing insulin secretion. Surf4 forms an oligomer and can physically interact with proinsulin and Sec12, essential for COPII vesicle formation. Our findings suggest that Surf4 interacts with proinsulin and delivers it into COPII vesicles for ER export in co-operation with Sec12 and COPII.


[Statistical and epidemiological studies on cases of infectious mononucleosis treated in the Infectious Diseases Division of the Livorno Hospitals between 1964 and 1977].
Reference is made to a series of 133 patients with infectious mononucleosis admitted to a hospital department over a period of 14 yr. The incidence of the disease is assessed in terms of sex, age, season, and occupation. Its duration and the average length of hospital stay are also computed. Particular attention is directed to 25 cases with a negative serum finding.


The development of integrated diabetes care in the Netherlands: a multiplayer self-assessment analysis.
Since recent years Dutch diabetes care has increasingly focused on improving the quality of care by introducing the concept of care groups (in Dutch: 'zorggroepen'), care pathways and improving cooperation with involved care professionals and patients. This study examined how participating actors in care groups assess the development of their diabetes services and the differences and similarities between different stakeholder groups. A self-evaluation study was performed within 36 diabetes care groups in the Netherlands. A web-based self-assessment instrument, based on the Development Model for Integrated Care (DMIC), was used to collect data among stakeholders of each care group. The DMIC defines nine clusters of integrated care and four phases of development. Statistical analysis was used to analyze the data. Respondents indicated that the diabetes care groups work together in well-organized multidisciplinary teams and there is clarity about one another's expertise, roles and tasks. The care groups can still develop on elements related to the management and monitoring of performance, quality of care and patient-centeredness. The results show differences (p < 0.01) between three stakeholders groups in how they assess their integrated care services; (1) core players, (2) managers/directors/coordinators and (3) players at a distance. Managers, directors and coordinators assessed more implemented integrated care activities than the other two stakeholder groups. This stakeholder group also placed their care groups in a further phase of development. Players at a distance assessed significantly less present elements and assessed their care group as less developed. The results show a significant difference between stakeholder groups in the assessment of diabetes care practices. This reflects that the professional disciplines and the roles of stakeholders influence the way they asses the development of their integrated care setting, or that certain stakeholder groups could be less involved or informed.


Osteochondromatosis with high concentration of procollagen II C peptide in joint fluid.
Procollagen II C peptide is used as a joint fluid marker of collagen synthesis by chondrocytes in patients with osteoarthritis. Since osteochondromatosis activates ectopic chondrogenesis in the synovial membrane, procollagen II C peptide is expected to be a good marker of the disease. Procollagen II C peptide can be used to evaluate the condition of the disease metabolically by simply aspirating the joint fluid. We have experienced a case of osteochondromatosis (Miligram's stage 2) with a high concentration of procollagen II C peptide. This case suggests that the concentration of procollagen II C peptide is a good marker of chondrogenesis by osteochondromatosis.


Integrated food chain surveillance system for Salmonella spp. in Mexico.
Few developing countries have foodborne pathogen surveillance systems, and none of these integrates data from humans, food, and animals. We describe the implementation of a 4-state, integrated food chain surveillance system (IFCS) for Salmonella spp. in Mexico. Significant findings were 1) high rates of meat contamination (21.3%-36.4%), 2) high rates of ceftriaxone-resistant S. Typhimurium in chicken, ill humans, and swine (77.3%, 66.3%, and 40.4% of S. Typhimurium T isolates, respectively), and 3) the emergence of ciprofloxacin resistance in S. Heidelberg (10.4%) and S. Typhimurium (1.7%) from swine. A strong association between Salmonella spp. contamination in beef and asymptomatic Salmonella spp. infection was only observed in the state with the lowest poverty level (Pearson r = 0.91, p<0.001). Pulsed-field gel electrophoresis analysis of 311 S. Typhimurium isolates showed 14 clusters with 102 human, retail meat, and food-animal isolates with indistinguishable patterns. An IFCS is technically and economically feasible in developing countries and can effectively identify major public health priorities.


Prefrontal activation during simulated driving in people with schizophrenia: A functional near-infrared spectroscopy study.
People with schizophrenia (PWS) could be at risk when driving, yet this remains to be confirmed. In this study, we used functional near-infrared spectroscopy (fNIRS) and a driving simulator to assess potential driving skill difficulties as reflected by brain activity in PWS and compared them with those of healthy controls (HCs). Twenty PWS and 20 HCs were evaluated. Four tasks were performed: 50-kph and 100-kph sudden braking and 50-kph left and right curve tasks. The hemodynamic activity and driving performance of the two groups were compared. No significant differences were found in the performance of the four tasks. However, significant differences in hemodynamic activity were observed in the left and right dorsolateral prefrontal cortex (DLPFC) during the 100-kph sudden braking task. In addition, a significant negative correlation was found between brake reaction time and brain activity in the left DLPFC during the 100-kph sudden braking task in both groups. The brain mechanisms involved in processing the mental load associated with driving a car are possibly similar in PWS and HCs. Our results suggest that PWS may be able to drive their vehicles safely in the community.


Photohydration of testosterone and 4-androstene-3,17-dione in aqueous solution.
Irradiation of testosterone, 4-androstene-3,17-dione, or their "half-molecule" 4,4a,5,6,7,8-hexahydro-4a-methyl-2(3H)-naphthalenone in dilute aqueous solutions with ultraviolet light of 254 nm wavelength caused rapid addition of water across the olefinic bond with formation of 5,17 beta-dihydroxy-5 alpha-androstan-3-one, 5-hydroxy-5 alpha-androstane-3-17-dione, and 9-hydroxy-10-methyl-2-decalone, respectively. Time-lapse spectrometry in the ultraviolet region showed that the photohydration of the androgenic steroids was extremely efficient and virtually free of the side reactions. Preparative photolytic reactions carried out in water-methanol solutions allowed isolation and characterization of photoproducts.


Associations between Intensity, Frequency, Duration, and Volume of Physical Activity and the Risk of Stroke in Middle- and Older-Aged Chinese People: A Cross-Sectional Study.
Persuasive evidence has shown the inverse associations between physical activity (PA) and the risk of stroke. However, few studies have investigated the associations between different dimensions (intensity, frequency, duration, volume) of PA and the risk of stroke. To investigate the associations between different dimensions of PA and the risk of stroke in total participants and subgroups. This study included 6250 individuals aged 45 years old and above from the China Health and Retirement Longitudinal Study (CHARLS). PA was divided into vigorous PA (VPA), moderate PA (MPA), and light PA (LPA), and described in different dimensions (intensity, frequency, duration, volume). Stroke was defined on the basis of self-reported diagnosis and related treatments. Binary logistic regression models were established to assess the associations between different dimensions of PA and the risk of stroke in total participants and subgroups stratified by sex. Individuals taking VPA with a frequency of 3-5 d/w, duration of ≥240 min/d, volume of ≥300 min/w had lower risks of strokes in total participants (Odds ratio (OR) = 0.32, 95% confidence interval (CI): 0.13, 0.75; OR = 0.60, 95% CI: 0.38, 0.94; OR = 0.68, 95% CI: 0.46, 0.99, respectively). However, significant associations of VPA with the risk of stroke in men were only observed in the duration of ≥240 min/d and volume of ≥300 min/w (OR = 0.53, 95% CI: 0.30, 0.93; OR = 0.61, 95% CI: 0.38, 0.99, respectively) whereas no significance in women. Compared with individuals taking no MPA, inverse significant associations between the risk of stroke and any level of frequency, duration and volume in MPA were observed in total sample (OR ranging from 0.16-0.40, all p < 0.05), whereas significant associations between the risk of stroke and MPA were found in men except the duration of 10-29 min/d and volume of 150-299 min/w (OR ranging from 0.26-0.35, all p < 0.05), and in women except the frequency of 1-2 d/w and duration of ≥240 min/d (OR ranging from 0.14-0.49, all p < 0.05). No significant associations could be observed in total participants and subgroups between LPA and the risk of stroke. This study revealed some significant associations between different dimensions of PA, especially MPA, and the risk of stroke. Furthermore, the difference of association was observed in the groups with different sex. Further prospective study is needed to determine deeper associations between PA and the risk of stroke.


Relapsing polychondritis.
Relapsing polychondritis (RP) is a rare systemic autoimmune disease characterized by episodic, progressive inflammatory destruction of cartilage. It can occur as an overlap syndrome in patients with other rheumatologic conditions. The disease usually follows an indolent relapsing-remitting course, but occasionally it can progress rapidly and even cause death. Although auricular or nasal chondritis or peripheral arthritis without other significant organ involvement are usually treated with low-dose corticosteroids, other more severe disease manifestations may require treatment with high-dose corticosteroids or other immunosuppressive agents. Biological targeted therapies might prove to be effective treatments of this condition.


Splenic lymphocyte production of an endorphin during endotoxic shock.
Endogenous opioids have been reported to elicit some of the pathophysiologic responses to endotoxic shock by binding to the delta-opiate receptor. We have previously reported the production of immunoreactive (ir)-endorphin by B lymphocytes treated with bacterial lipopolysaccharide (LPS). We postulated that this lymphocyte-derived ir-endorphin may be an extrapituitary source of the endogenous opioid component associated with the pathophysiology of endotoxic shock. To test this hypothesis, we chose to study the LPS-sensitive (C3HeB/FeJ) and -resistant (C3H/HeJ) inbred mouse model. We treated these mice with intraperitoneal injections of LPS or B-lymphocyte-derived ir-endorphin. The LPS-sensitive mice presented with a severe hypothermic and pathophysiologic response pattern when treated with LPS or with ir-endorphin. The LPS-resistant mice, which were unresponsive to the LPS, however, presented with the typical hypothermic and pathophysiologic responses to the ir-endorphin. Immunofluorescence on the splenic leukocytes in the LPS-treated mice showed significant ir-endorphin present only in the LPS-sensitive mice at a time point preceding onset of the pathophysiologic response pattern. Taken together, this evidence strongly suggests a role for B-lymphocyte-derived ir-endorphin in the pathophysiology of endotoxic shock. The implications of immune system regulation of neuroendocrine function are discussed.


Identification and characterization of an aβ oligomer precipitating peptide that may be useful to explore gene therapeutic approaches to Alzheimer disease.
A key feature of Alzheimer disease (AD) is the pathologic self-association of the amyloid-β (Aβ) peptide, leading to the formation of diffusible toxic Aβ oligomers and extracellular amyloid plaques. Next to extracellular Aβ, intraneuronal Aβ has important pathological functions in AD. Agents that specifically interfere with the oligomerization processes either outside or inside of neurons are highly desired for the elucidation of the pathologic mechanisms of AD and might even pave the way for new AD gene therapeutic approaches. Here, we characterize the Aβ binding peptide L3 and its influence on Aβ oligomerization in vitro. Preliminary studies in cell culture demonstrate that stably expressed L3 reduces cell toxicity of externally added Aβ in neuroblastoma cells.


Therapeutic cloning: the ethical limits.
A brief outline of stem cells, stem cell therapy and therapeutic cloning is given. The position of therapeutic cloning with regard to other embryonic manipulations - IVF-based reproduction, embryonic stem formation from IVF embryos and reproductive cloning - is indicated. The main ethically challenging stages in therapeutic cloning are considered to be the nuclear transfer process including the source of eggs for this and the destruction of an embryo to provide stem cells for therapeutic use. The extremely polarised nature of the debate regarding the status of an early human embryo is noted, and some potential alternative strategies for preparing immunocompatible pluripotent stem cells are indicated.


Effects of omecamtiv mecarbil and mavacamten in isolated human atrium.
Heart failure is a syndrome that can result from impaired heart muscle contractions like in dilative cardiomyopathy but also from hypertrophic obstructive cardiomyopathy (HOCOM). A pharmacological therapy might lie in Ca2+-sensitizing or Ca2+-desensitizing drugs, respectively. Such drugs are thought to be omecamtiv mecarbil (OME) and mavacamten (MYK-461), respectively. Their function in contracting human muscle is not fully understood and was the focus of the present study. OME from 1 nM to 10 µM cumulatively applied failed to raise force of contraction in human right atrial preparations strips (HAP) or mouse left atrial preparations (LA). However, OME prolonged time to peak tension and time of relaxation in HAP and LA but did not alter the beating rate in right atrial preparations from mice (RA). In contrast, MYK-461 (10 nM to 10 µM) reduced concentration- and time-dependently force of contraction in HAP and LA. MYK-461 (10 µM) did not affect the beating rate in RA. In summary, the present data failed to detect an increase in force of contraction for OME, in human and mouse atrium. In contrast, a Ca2+ desensitizer studied for comparison was able to reduce force of contraction in HAP and LA. We conclude that putative beneficial effects of OME in dilated cardiomyopathy cannot be explained by positive inotropic effects in the HAP, whereas beneficial functional effects of MYK-461 in HOCOM can be explained by negative inotropic effects in HAP.


Increased digitoxin cleavage by liver microsomes of spironolactone-pretreated rats.
Pretreatment of rats with spironolactone caused an fourfold increased cleavage rate of the sugar chain of digitoxin (dt-3) in vitro yielding digitoxigenin-bis-digitoxoside. This was due to an enhanced, cyt. P450 dependent, formation of 15'-dehydro-dt-3, the intermediate which has to be formed before the terminal sugar can be split off. The second reaction catalysed by microsomal monoxygenases, the 12-beta-hydroxylation, was only increased by a factor 2. In contrast to the effects of spironolactone no increase of metabolism could be observed after phenobarbital pretreatment. From our results it may be concluded that the enhanced dt-3 metabolism in vivo is mainly caused by spironolactone inducible monoxygenases which catalyse the oxidation of the terminal sugar.


Strong and flexible lignocellulosic film fabricated via a feasible molecular remodeling strategy.
Biomass-derived sustainable film is a promising alternative to synthetic plastic, but hampered by strength, toughness and flexibility trade-off predicament. Here, a feasible and scalable strategy was proposed to fabricate strong and flexible lignocellulosic film through molecular reconstruction of cellulose and lignin. In this strategy, polyphenol lignin was absorbed and wrapped on the surface of cellulose fiber, forming strong interfacial adhesion and cohesion via intramolecular and intermolecular hydrogen bonding. Further, covalent ether bond was generated between the hydroxyl groups of lignocellulose to form chemical cross-linking network induced by epichlorohydrin (ECH). The synergistic effect of hydrogen bonding and stable chemical cross-linking enabled the resultant lignocellulosic film (ELCF) with outstanding mechanical strength of 132.48 MPa, the elongation at break of 9.77 %, and toughness of 9.77 MJ·m-3. Notably, the integration of polyphenol lignin synergistically improved the thermal stability, water resistance, UV-blocking performances of ELCF. Importantly, after immersion for 30 d, ELCF still possessed high wet strength of 70.38 MPa, and elongation at break of 7.70 %, suggesting excellent and durable mechanical performances. Moreover, ELCF could be biodegraded in the natural soil. Therefore, this study provides a new and versatile approach to reconstruct highly-performance lignocellulosic films coupling strength, toughness with flexibility for promising plastic replacement.


The use of debonding pliers in the removal of ceramic brackets: force levels and enamel cracks.
The purpose of this study was to determine the magnitude of the applied forces when using pliers as a debonding method for removing three types of ceramic brackets. The following variables were evaluated: debonding strengths, the Adhesive Remnant Index, and enamel damage with transillumination. All the brackets in this investigation was debonded with a direct bond remover plier. An Instron Model Universal testing machine was used to measure debonding strengths. From the results of this study, the following conclusions can be drawn: (1) The diamteral compression mean bond strengths for the three bracket types tested ranged from a low of 67.8 Kg/cm2 to a high of 102.9 Kg/cm2 and are in the clinically acceptable range for orthodontic force application. (2) There was no significant difference in the mean bond strengths of the three ceramic brackets tested. (3) The ARI scores were found to range between 2 and 4 indicating a cohesive type of bond failure. (4) Transillumination was used to evaluate minute enamel damage, and the results indicated that most of the teeth (82.02%) experienced no increase in enamel cracks after debonding. The teeth that showed an increase in the number of cracks after debonding had a significantly higher mean bond strength (113.0 Kg/cm2) than those with no increase in the number of cracks (72.9 Kg/cm2).


Genetic structure and polymorphisms of the N16 gene in Pinctada fucata.
The molluscan shell is a composite of inorganic crystals comprising calcium carbonate and a minute amount of organic matrix. The organic matrix (OM) is intimately involved in every step of shell formation and has consequently received much attention in recent years. However, most of the deposited information has resulted from cDNA analysis, with little analysis of the genome, including the presence and effects of polymorphic genes encoding OM proteins. The current study aimed to clarify the genome structure of the N16 gene from the Japanese pearl oyster, Pinctada fucata, with particular reference to polymorphisms. The N16 gene was analyzed using PCR and DNA sequencing. 23 polymorphic variants were identified from 28 individuals. The variants were analyzed for their relationship to shell formation. All the variations detected by genomic PCR appeared in the cDNAs, implying that all the polymorphisms were transcribed and translated into N16 proteins. Additional genome analysis revealed at least two N16 genes, which were sequentially positioned, each of them comprising four exons and three introns. Further analyses of the transcriptional regulation and function of the N16 genes may provide new insights into their role in molluscan biomineralization.


HIV viral load: a concept analysis and critique.
In this article, we examine the concept of HIV viral load and how it has evolved over time (1995-2013) in the field of HIV/AIDS. Although the term viral load is used extensively in this field, few efforts have been directed toward the conceptualization of HIV viral load, which is often left unquestioned, undertheorized, and portrayed as a neutral and objective laboratory value that has remained relatively stable over time--with the exception of progressive advancements in technology, techniques, and sensitivity. The purpose of this article is to apply the evolutionary concept analysis method developed by Rodgers (1989, 2000a) to the concept of HIV viral load. To set the stage, we establish the need for a concept analysis of HIV viral load and provide an overview of the evolutionary view. Then, drawing on the steps proposed by Rodgers (2000a), we outline the process of data collection, management, and analysis. We then offer an in-depth discussion of the findings (attributes, antecedents, and consequences) informed by Wuest's (2000) critical approach to concept analysis. We conclude by highlighting the implications of this analysis for clinical practice, research, and theory.


Actinomycosis of the middle ear.
Actinomycosis is an uncommon infection of the middle ear. Only 21 cases of actinomycosis of the middle ear have been reported in the English literature prior to this paper. The offending organism is Actinomyces israelii, which is an anaerobic, filamentous organism that is difficult to grow in culture. The infection is chronic and is seldom diagnosed prior to tympanomastoidectomy. The identification of small, yellow, glue-like masses, which are called sulfur granules, is often the key to making the diagnosis of actinomycosis of the middle ear. Following tympanomastoidectomy, penicillin is given orally for 3-6 months.


Epidermodysplasia verruciformis in Africans.
Epidermodysplasia verruciformis (EV) is a rare cutaneous disorder characterised by persistent, refractory infection with human papillomaviruses (HPV). Although EV does not seem to have racial or geographic preference, there is a scarcity of reports on its occurrence in Africans. Twenty Africans with EV were studied, and the literature on this condition in Africans was reviewed. Virologic studies were performed on 10 patients. Three types of lesions were observed: flat warts, pityriasis versicolor-like macules, and seborrheic keratosis-like changes. Malignant transformation occurred in only one patient. HPV-3 was isolated only from flat warts, HPV-5 and HPV-17 were isolated only from PV-like lesions, whereas an HPV-5-related type was found in all three types of changes. HPV-5-related type revealed DNA that was related but not identical to any of the viruses in the HPV-5 group. This particular type was isolated from all five South African patients with EV in whom virologic studies were performed. The benign nature of EV in dark-skinned Africans has been confirmed. Four HPV types have been isolated, of which HPV-related type was found in all South African patients with EV and in all types of skin changes, regardless of their morphology. African patients with EV frequently present seborrheic keratosis-like changes.


Knee arthroscopy has limited effects on relieving local symptoms of knee osteoarthritis: an analysis of data from the Osteoarthritis Initiative.
Knee arthroscopy's efficacy in symptom improvement for knee osteoarthritis remains debated. In this study, we analyzed a multicenter database to investigate local symptom improvement. We extracted and analyzed the data of 163 patients from the Osteoarthritis Initiative cohort who underwent unilateral knee arthroscopy (UKA) and were followed up for at least 24 months. UKA patients were matched to non-UKA patients (n = 163) according to sex, age, abdominal circumference, and Kellgren-Lawrence grade. The verified KOOS questionnaires (knee catching, locking, grinding, or clicking) and common local symptoms (frequent knee pain, aching, or stiffness) were set as outcomes. Furthermore, we built a binary logistic regression model to examine the relationship between UKA and local symptom improvement and new-onset symptoms, adjusting for conservative therapeutic covariables (injection of steroids or transparent acid into the knee joint, oral chondroitin sulfate, amino glucose, or analgesics). Analysis showed that the UKA and non-UKA groups showed no obvious difference in the three knee symptoms, but the probability of new-onset grinding or clicking, and frequent knee pain, aching, or stiffness symptoms in the UKA group were respectively 5.82 and 5.65-fold higher than that in the non-UKA group. After analyzing conservative treatment data using a multiple imputation method, the results were consistent with previous regression analyses. Compared to the non-UKA group, the UKA group showed no noticeable differences in the improvement of the three knee symptoms and showed an increased the probability of new-onset grinding or clicking and frequent knee pain, aching, or stiffness symptoms. Key Points • Knee arthroscopy may increase the probability of new-onset grinding or clicking and frequent knee pain, aching, or stiffness symptoms. • We found no difference in the improvement of local knee symptoms (knee catching, locking, grinding, clicking or frequent pain, aching, or stiffness) improvement between the two groups with or without knee arthroscopy.