Pseudo 2-fold symmetry in the copper-binding domain of arthropodan haemocyanins. Possible implications for the evolution of oxygen transport proteins.
Investigation of the copper-binding centre of Panulirus interruptus haemocyanin led to the discovery of a pseudo 2-fold axis relating two helical pairs surrounding and co-ordinating the two copper ions. The pseudo 2-fold symmetry relating one helical pair, co-ordinating Cu-A, to the second helical pair co-ordinating Cu-B is quite precise with 31 equivalent C alpha atoms having a root-mean-square deviation of only 1.47 A. The 2-fold consists of a rotation of 174.6 degrees and a translation parallel to the rotation axis of 0.7 A. After superposition of the helical pairs, the two copper ions are within 1.1 A and the three C alpha atoms of the histidine ligands of Cu-A are within a root-mean-square deviation of 1.0 A from the C alpha atoms of the histidine residues co-ordinating Cu-B. Of the superimposed residues, 26% are identical in sequence. These data suggest that the current oxygen-binding centre of arthropodan haemocyanins is the result of dimerization, gene duplication and gene fusion of an ancestral mono-copper-binding helical pair. This suggestion is supported by the recent discovery that in the sequence of functional domains of molluscan haemocyanins only amino acid sequence homology with the arthropodan Cu-B helical pair has been found and no evidence for similarity with a Cu-A binding helical pair was observed. This provides strong evidence that a mono-copper-binding helical pair has been the ancestor of both the arthropodan and molluscan haemocyanins. Turning to the Fe-binding helical pairs in haemerythrins, it appears that they are less similar to each other than the two Cu-binding helical pairs in arthropodan haemocyanins. Nevertheless, the Fe-B haemerythrin helical pair superimposes well onto the Cu-A helical pair of Panulirus haemocyanin. A root-mean-square deviation of 1.9 A for 24 equivalent C alpha carbon atoms is obtained, while Fe-B deviates 1.4 A from Cu-A after superposition of the helices. Moreover, the three histidine ligands of the Cu-A helical pair are equivalent with three histidine ligands of the Fe-B pair. The structural similarity and correspondence in metal-binding ligands suggests that both haemocyanins and haemerythrins have originated from an ancestral mono-metal-binding helical pair having two ligands provided by the first helix and one ligand by the second helix.(ABSTRACT TRUNCATED AT 400 WORDS)


Geriatric assessment prior to cancer treatment: A health economic evaluation.
To address uncertainty regarding the cost-effectiveness of implementing geriatric assessment (GA) in oncology practice, we undertook a synthetic, model-based economic evaluation. A decision-analytic model with embedded Markov chains was developed to simulate a cost-effectiveness analysis of implementing GA within standard oncological care compared to current practice. This was for patients aged 77 years (the mean age in included trials) receiving chemotherapy or surgery as first-line treatment. Assumptions were made about model parameters, based on available literature, to calculate the incremental net health benefit (INHB) of GA, using a data synthesis. GA has additional costs over standard care alone of between £390 and £576, depending upon implementation configuration. When major assumptions about the effectiveness of GA were modelled, INHB was marginally positive (0.09-0.12) at all cost-effectiveness thresholds (CETs). If no reduction in postoperative complications was assumed, the intervention was shown not to be cost-effective (INHB negative at all CETs). When used before chemotherapy, with minimal healthcare staffing inputs and technological assistance, GA is cost-effective (INHB positive between 0.06 and 0.07 at all CETs). Considering emerging evidence that GA improves outcomes in oncology, GA may not be a cost-effective intervention when used for all older adults with cancer. However, with judicious selection of implementation models, GA has the potential to be cost-effective. Due to significant heterogeneity and centre dependent success in implementation and effectiveness, GA is difficult to study in oncology settings. Stakeholders could take a pragmatic approach towards GA introduction with local evaluation favoured over generalisable research. Because GA tends towards utilitarianism and has no safety issues, it is a suitable intervention for more widespread implementation.


An evaluation of self-reported mobile phone use compared to billing records among a group of engineers and scientists.
Most epidemiologic studies of potential health impacts of mobile phones rely on self-reported information, which can lead to exposure misclassification. We compared self-reported questionnaire data among 60 participants, and phone billing records over a 3-year period (2002-2004). Phone usage information was compared by the calculation of the mean and median number of calls and duration of use, as well as correlation coefficients and associated P-values. Average call duration from self-reports was slightly lower than billing records (2.1 min vs. 2.8 min, P = 0.01). Participants reported a higher number of average daily calls than billing records (7.9 vs. 4.1, P = 0.002). Correlation coefficients for average minutes per day of mobile phone use and average number of calls per day were relatively high (R = 0.71 and 0.69, respectively, P < 0.001). Information reported at the monthly level tended to be more accurate than estimates of weekly or daily use. Our findings of modest correlations between self-reported mobile phone usage and billing records and substantial variability in recall are consistent with previous studies. However, the direction of over- and under-reporting was not consistent with previous research. We did not observe increased variability over longer periods of recall or a pattern of lower accuracy among older age groups compared with younger groups. Study limitations included a relatively small sample size, low participation rates, and potential limited generalizability. The variability within studies and non-uniformity across studies indicates that estimation of the frequency and duration of phone use by questionnaires should be supplemented with subscriber records whenever practical.


Role of stem design and material on stress distributions in cemented total hip replacement.
The risk of fatigue fractures of the femoral stem in a cemented total hip arthroplasty can be minimized by either increasing the stem cross-section and/or using a very high strength alloy. The object of this study was to compare important mechanical characteristics of five selected stem designs, differing in configuration and material (stainless steel, cast chrome cobalt alloy, nickel based alloy and titanium alloy). The stain pattern on the stem was analysed in a 3-point-bending jig and also after cementing it into cadaver femurs. Regardless of stem type or test method, the typical tensile stress distribution on the lateral stem was a bell shaped curve. For the cobalt-chrome and stainless steel stems, the larger the stem the lower were the stem stresses and the stress gradient, and the higher was the factor of safety. However, the factor of safety was increased even further by the use of super alloys such as MP35N and Ti6Al4V. In addition, Ti6Al4V alloy allowed the use of larger and stronger stems without the extra penalty of rigidity, which was enforced by either the steel or cobalt based alloy.


'This is uncharted water for all of us': challenges anticipated by hospital clinicians when voluntary assisted dying becomes legal in Victoria.
Objective The aim of this study was to identify the challenges anticipated by clinical staff in two Melbourne health services in relation to the legalisation of voluntary assisted dying in Victoria, Australia. Methods A qualitative approach was used to investigate perceived challenges for clinicians. Data were collected after the law had passed but before the start date for voluntary assisted dying in Victoria. This work is part of a larger mixed-methods anonymous online survey about Victorian clinicians' views on voluntary assisted dying. Five open-ended questions were included in order to gather text data from a large number of clinicians in diverse roles. Participants included medical, nursing and allied health staff from two services, one a metropolitan tertiary referral health service (Service 1) and the other a major metropolitan health service (Service 2). The data were analysed thematically using qualitative description. Results In all, 1086 staff provided responses to one or more qualitative questions: 774 from Service 1 and 312 from Service 2. Clinicians anticipated a range of challenges, which included burdens for staff, such as emotional toll, workload and increased conflict with colleagues, patients and families. Challenges regarding organisational culture, the logistics of delivering voluntary assisted dying under the specific Victorian law and how voluntary assisted dying would fit within the hospital's overall work were also raised. Conclusions The legalisation of voluntary assisted dying is anticipated to create a range of challenges for all types of clinicians in the hospital setting. Clinicians identified challenges both at the individual and system levels. What is known about the topic? Voluntary assisted dying became legal in Victoria on 19 June 2019 under the Voluntary Assisted Dying Act 2017. However there has been little Victorian data to inform implementation. What does this paper add? Victorian hospital clinicians anticipate challenges at the individual and system levels, and across all clinical disciplines. These challenges include increased conflict, emotional burden and workload. Clinicians report concerns about organisational culture, the logistics of delivering voluntary assisted dying under the specific Victorian law and effects on hospitals' overall work. What are the implications for practitioners? Careful attention to the breadth of staff affected, alongside appropriate resourcing, will be needed to support clinicians in the context of this legislative change.


XAF1 prevents hyperproduction of type I interferon upon viral infection by targeting IRF7.
Interferon regulatory factor (IRF) 3 and IRF7 are master regulators of type I interferon (IFN-I)-dependent antiviral innate immunity. Upon viral infection, a positive feedback loop is formed, wherein IRF7 promotes further induction of IFN-I in the later stage. Thus, it is critical to maintain a suitably low level of IRF7 to avoid the hyperproduction of IFN-I. In this study, we find that early expression of IFN-I-dependent STAT1 promotes the expression of XAF1 and that XAF1 is associated specifically with IRF7 and inhibits the activity of XIAP. XAF1-knockout and XIAP-transgenic mice display resistance to viral infection, and this resistance is accompanied by increases in IFN-I production and IRF7 stability. Mechanistically, we find that the XAF1-XIAP axis controls the activity of KLHL22, an adaptor of the BTB-CUL3-RBX1 E3 ligase complex through a ubiquitin-dependent pathway. CUL3-KLHL22 directly targets IRF7 and catalyzes its K48-linked ubiquitination and proteasomal degradation. These findings reveal unexpected functions of the XAF1-XIAP axis and KLHL22 in the regulation of IRF7 stability and highlight an important target for antiviral innate immunity.


Heparin removal after cardiopulmonary bypass in a patient with adverse reaction to protamine.
A 74-year-old man underwent elective coronary surgery under cardiopulmonary bypass. A few minutes after the protamine administration was started, he suddenly developed a severe hypotension necessitating cardiac massage and recannulation for pump assistance. A further test dose of protamine provoked an identical reaction. We installed a Heparin Removal Device, which allows for ex-vivo deheparinization. In 35 minutes ACT decreased from 480 sec to 180 sec and clots appeared in the operating field. This system provides an excellent alternative to protamine in patients with an adverse reaction to protamine.


The in vivo and in vitro effects of rhG-CSF on allergic, haematological and biochemical variables.
The objective was to evaluate the influence of treatment with rhG-CSF on allergic indexes. This was done by open trial of 5 days' treatment with rhG-CSF (5 microg/kg/day s. c.). 10 patients (6 men), aged 28 to 54 years, with rhinoconjunctivitis due to grass pollen allergy, participated in the investigation. Main measures were blood count, basophil histamine release, skin test and conjunctival provocation test. The treatment resulted in significant increases in numbers of neutrophils (590%), basophils (280%), eosinophils (250%) and lymphocytes (71%). Total blood histamine was increased, but basophil histamine releasability was decreased. Serum alkaline phosphatase increased 92% and serum lactate dehydrogenase increased 35% (both significant). There were no significant changes in the skin tests and the conjunctival provocation tests. Two months after the treatment all tests had returned to baseline levels. Five of the patients (50%) reported side effects, one withdrew. In conclusion treatment with rhG-CSF increases the number of circulating blood cells other than neutrophils without causing changes in indexes of allergic reactivity.


DNA methylation-based biomarkers for early detection of non-small cell lung cancer: an update.
Lung cancer is the number one cancer killer in the United States. This disease is clinically divided into two sub-types, small cell lung cancer, (10-15% of lung cancer cases), and non-small cell lung cancer (NSCLC; 85-90% of cases). Early detection of NSCLC, which is the more common and less aggressive of the two sub-types, has the highest potential for saving lives. As yet, no routine screening method that enables early detection exists, and this is a key factor in the high mortality rate of this disease. Imaging and cytology-based screening strategies have been employed for early detection, and while some are sensitive, none have been demonstrated to reduce lung cancer mortality. However, mortality might be reduced by developing specific molecular markers that can complement imaging techniques. DNA methylation has emerged as a highly promising biomarker and is being actively studied in multiple cancers. The analysis of DNA methylation-based biomarkers is rapidly advancing, and a large number of potential biomarkers have been identified. Here we present a detailed review of the literature, focusing on DNA methylation-based markers developed using primary NSCLC tissue. Viable markers for clinical diagnosis must be detectable in 'remote media' such as blood, sputum, bronchoalveolar lavage, or even exhaled breath condensate. We discuss progress on their detection in such media and the sensitivity and specificity of the molecular marker panels identified to date. Lastly, we look to future advancements that will be made possible with the interrogation of the epigenome.


Three dimensionality of gleno-humeral deformities in obstetrical brachial plexus palsy.
The primary objective of this study was to test the hypothesis that gleno-humeral deformity in children and adolescent with obstetrical brachial plexus palsy is three-dimensional (3D). The study also compared the metrological properties of typical two-dimensional gleno-humeral measures to the newly developed 3D measures. Thirteen individuals (age = 11.8 ± 3.3 years) with obstetrical brachial plexus palsy participated in this IRB-approved study. 3D axial magnetic resonance images were acquired for both shoulders. Glenoid and humeral models were created in order to quantify 3D glenoid version, humeral head migration, and glenoid concavity. Two-dimensional (2D) measures were acquired as recommended in the literature. All measures were completed by two observers in this observer-blind study. Compared to the non-involved side, the glenoid was more retroverted (7.91°, p = 0.003) and inferiorly oriented (7.28°, p = 0.009). The humeral head was migrated more posteriorly (5.54 mm, p = 0.007), inferiorly (-3.96 mm, p = 0.013), and medially (-3.63 mm,p = 0.002). Eleven of the 13 glenoids were concave, based on the 3D glenoid models. The concurrent validity between three- and 2D measures were highly dependent of the parameter measured, the slice level used for the 2D analysis, and the presence/absence of pathology (0.63 < r < 0.91). The standard error of measurement for the 2D anterior-posterior version (>3°) was larger than that for the 3D measure of version (<1°) on the involved side. This study clearly demonstrated that the gleno-humeral deformation in obstetrical brachial plexus palsy is 3D, emphasizing the need for 3D subject specific gleno-humeral shape analysis for follow-up and treatment plans in children with obstetrical brachial plexus palsy.


Catecholamines in discrete areas of the hypothalamus of obese and castrated male rats.
Levels of norepinephrine (NE) and dopamine (DA) were measured in eight discrete regions of the hypothalamus in three groups of male rats; genetically obese (fafa), non-obese (FaFa) and castrated non-obese (FaFa). DA levels showed no significant differences among the groups in any of the regions. NE levels in the paraventricular nucleus (PVN) were significantly lower in the obese and castrated animals than in the normal animals. In the median eminence (ME), NE levels were significantly decreased for the castrated group. None of the other regions sampled showed significant differences in NE levels.


Editor's Choice - Comparison of the Reproducibility of Ultrasound Calliper Placement Methods in Abdominal Aortic Diameter Measurements: A Systematic Review and Meta-Analysis of Diagnostic Test Accuracy Studies.
To assess which ultrasound (US) method of maximum anteroposterior (AP) abdominal aortic diameter measurement can be considered most reproducible. MEDLINE, Scopus, and Web of Science were searched (PROSPERO ID: 276694). Eligible studies reported intra- and or interobserver agreement according to Bland-Altman analysis (mean ± standard deviation [SD]) for abdominal aortic diameter AP US evaluations with an outer to outer (OTO), inner to inner (ITI), and or leading edge to leading edge (LELE) calliper placement. The Preferred Reporting Items for a Systematic Review and Meta-Analysis of Diagnostic Test Accuracy Studies statement was followed. The QUADAS-2 tool and QUADAS-C extension were used for risk of bias assessment and the GRADE framework to rate the certainty of evidence. Pooled estimates (fixed effects meta-analysis, after a test of homogeneity of means) for each US method were compared with pairwise one sided t tests. Sensitivity analyses (for studies published in 2010 or later) and meta-regression were also performed. 21 studies were included in the qualitative analysis. Twelve were eligible for quantitative analysis. Studies showed heterogeneity in the US model and transducer used, sex of participants, and observer professions, expertise, and training. Included studies shared a common mean for each US method (OTO: p = 1.0, ITI: p = 1.0, and LELE: p = 1.0). A pooled estimate of interobserver reproducibility for each US method was obtained, combining the mean ± SD (Bland-Altman analysis) from each study: OTO: 0.182 ± 0.440; ITI: 0.170 ± 0.554; and LELE: 0.437 ± 0.419. There were no statistically significant differences between the methods (OTO vs. ITI: p = .52, OTO vs. LELE: p = .069, ITI vs. LELE: p = .17). Considering studies published in 2010 and later, the pooled estimate for LELE was the smallest, without statistically significant differences between the methods. Despite the low risk of bias, the certainty of the evidence for both meta-analysed outcomes remained low. The interobserver reproducibility for OTO and ITI was 2.5 times smaller (indicating better reproducibility) than LELE; however, without statistically significant differences between the methods and low GRADE evidence certainty. Additional data are needed to validate these findings, while inherent differences between the methods need to be emphasised.


Alterations in morning cortisol associated with PTSD in women with breast cancer.
Diagnosis and treatment of breast cancer can be a stressful experience, putting women at risk of posttraumatic stress disorder (PTSD). The current study investigated morning cortisol levels in newly diagnosed (i.e., within 6 months) breast cancer patients. Structured DSM-IV interviews determined current and past incidence of PTSD and major depressive disorder (MDD) in 71 women with Stage 0-3 breast cancer. Significantly decreased plasma cortisol was found in women meeting current or lifetime criteria for PTSD or past diagnosis of MDD. These results reinforce the importance for both psychological and physiological outcomes of a clinical evaluation of both current and past psychiatric status in newly diagnosed cancer patients.


Synthesis and structure-activity relationships of novel indirubin derivatives as potent anti-proliferative agents with CDK2 inhibitory activities.
Indirubin, an active ingredient of a traditional Chinese recipe Danggui Longhui Wan, has been known as a CDK inhibitor competing with ATP for binding to the catalytic site of cyclin-dependent kinases (CDKs). Since CDKs, a group of serine/threonine kinases forming active heterodimeric complexes with cyclins, are key regulators of the cell cycle regulation, therapeutic interventions targeting CDKs have been stimulated for the treatment of proliferative diseases, such as cancer, psoriasis, and for the prevention of chemotherapy-associated side effects, such as alopecia. A series of novel indirubin analogs was synthesized and evaluated for anti-proliferative and CDK2 inhibitory activities. Among the indirubin derivatives tested in the growth inhibitions against several human cancer cell lines, 5-nitro, halide, and bulky group containing acylamino substituted analogs showed high anti-proliferative effects. Selected analogs showing potent anti-proliferative activities were evaluated further in the CDK2 enzyme assay, which resulted in the discovery of potent CDK2 inhibitors.


Inhibitory effects of berberine on lipopolysaccharide-induced inducible nitric oxide synthase and the high-mobility group box 1 release in macrophages.
We investigated the molecular mechanism by which berberine reduces nitric oxide (NO) expression and high-mobility group box 1 (HMGB1) release in lipopolysaccharide (LPS)-induced macrophages. Berberine significantly inhibited the LPS-stimulated NO production and HMGB1 release in macrophages. In addition, berberine also induced heme oxygenase (HO)-1 in a dose-dependent manner, which was mediated through activation of p38 MAPK and NF-E2-related factor 2 (Nrf2) signaling cascade in macrophages. The inhibitory effect of berberine on LPS-stimulated NO and HMGB1 release was reversed by siRNA-Nrf2, SB203580 (p38 MAPK inhibitor) and zinc protoporphyrin (ZnPP; HO-1 inhibitor) within macrophages. Therefore, we conclude that berberine inhibits the proinflammatory response to LPS in macrophages by up-regulation of the HO-1 level, in which p38 MAPK and Nrf2 have an important role. These results suggest that berberine may be useful as a therapeutic agent for the treatment of inflammatory diseases.


Candidate editases for GluR channels in single neurons of rat hippocampus and cerebellum.
RNA editing by site selective adenosine deamination changes codons in several nuclear transcripts in the mammalian brain and affects critical properties of the encoded proteins, as exemplified by the calcium permeability of AMPA receptor channels. The recently cloned RNA dependent adenosine deaminases ADAR1, ADAR2 and ADAR3 form a small family of sequence-related candidate editases which are expressed in brain and other tissues at distinct levels and patterns. We have employed single-cell polymerase chain reaction of hippocampal CA1 and CA3 pyramidal neurons and cerebellar Purkinje and Bergmann glial cells in an attempt to evaluate the expression of these enzymes at a cellular level. We found ADAR2 expressed in all cells analyzed; approximately 50% of the cells co-expressed ADAR1 or ADAR3. The differential ADAR expression revealed by our study might underlie the distinct editing efficiencies and selectivities in different GluR subunit transcripts.


Biosynthesis of glycoproteins in the pathogenic fungus Candida albicans: activation of dolichol phosphate mannose synthase by cAMP-mediated protein phosphorylation.
Following incubation with ATP and a cAMP-dependent protein kinase under optimal conditions of lipid acceptor, phospholipid and metal ion requirements, the transfer activity of partially purified dolichol phosphate mannose synthase (DPMS) increased about 60% and this activation correlated with a 50% increase in V(max) with no alteration in the apparent K(m) for GDP-Manose. Phosphorylation with [gamma-(32)P]ATP resulted in the labeling of several polypeptides, one of which exhibited the molecular weight of the enzyme (30 kDa) and was also recognized using a specific anti-DPMS monoclonal antibody. This and the fact that the phosphate label could be removed by an alkaline phosphatase indicate that Candida DPMS may be regulated by phosphorylation-dephosphorylation, a mechanism that has been proposed for the enzyme in other organisms.


FAM96A knock-out promotes alternative macrophage polarization and protects mice against sepsis.
Sepsis is an intractable clinical syndrome characterized by organ dysfunction when the body over-responds to an infection. Sepsis has a high fatality rate and lacks effective treatment. Family with sequence similarity 96 member A (FAM96A) is an evolutionarily conserved protein with high expression in the immune system and is related to cytosolic iron assembly and tumour suppression; however, research has been rarely conducted on its immune functions. Our study found that Fam96a-/- mice significantly resisted lesions during sepsis simulated by caecal ligation and puncture (CLP) or endotoxicosis models. After a challenge with lipopolysaccharide (LPS) or infection, Fam96a-/- mice exhibited less organ damage, longer survival and better bacterial clearance with decreased levels of proinflammatory cytokines. While screening several subsets of immune cells, FAM96A-expressing macrophages as the key cell type inhibited sepsis development. In-vivo macrophage depletion or adoptive transfer experiments abrogated significant differences in the survival of sepsis between Fam96a-/- and wild-type mice. Results of the bone marrow-derived macrophage (BMDM) polarization experiment indicated that FAM96A deficiency promotes the transformation of uncommitted monocytes/macrophages (M0) into M2 macrophages, secreting fewer proinflammatory cytokines. FAM96A may mediate an immunometabolism shift - from oxidative phosphorylation (OXPHOS) to glycolysis - in macrophages during sepsis, mirrored by reactive oxygen species (ROS) and glucose uptake. These data demonstrate that FAM96A regulates inflammatory response and provide a novel genomic insight for sepsis treatment.


Electrophysiological evidence of perceived sexual attractiveness for human female bodies varying in waist-to-hip ratio.
The dynamics of brain activation reflecting attractiveness in humans are unclear. Among the different features affecting attractiveness of the female body, the waist-to-hip ratio (WHR) is considered to be crucial. To date, however, no event-related potential (ERP) study has addressed the question of its associated pattern of brain activation. We carried out two different experiments: (a) a behavioural study, to judge the level of attractiveness of female realistic models depicting 4 different WHRs (0.6, 0.7, 0.8, 0.9) with and without clothes; (b) an EEG paradigm, to record brain activity while participants (heterosexual men and women) viewed these same models. Behavioural results showed that WHRs of 0.7 were considered more attractive than the others. ERP analyses revealed a different pattern of activation for male and female viewers. The 0.7 ratio elicited greater positivity at the P1 level in male viewers but not females. Naked bodies increased the N190 in both groups and peaked earlier for the 0.7 ratio in the male viewers. Finally, the late positive component (LPC) was found to be greater in male than in female viewers and was globally more marked for naked bodies as well as WHRs of 0.7 in both groups of viewers. These results provide the first electrophysiological evidence of specific time periods linked to the processing of a body feature denoting attractiveness and therefore playing a role in mate choice.


Simulation-based training of internal medicine residents in advanced cardiac life support protocols: a randomized trial.
Internal medicine residents must be competent in Advanced Cardiac Life Support (ACLS) for board certification. The purpose was to use a medical simulator to assess baseline proficiency in ACLS and determine the impact of an intervention on skill development. This was a randomized trial with wait-list controls. After baseline evaluation in all residents, the intervention group received 4 education sessions using a medical simulator. All residents were then retested. After crossover, the wait-list group received the intervention, and residents were tested again. Performance was assessed by comparison to American Heart Association guidelines for treatment of ACLS conditions with interrater and internal consistency reliability estimates. Performance improved significantly after simulator training. No improvement was detected as a function of clinical experience alone. The educational program was rated highly.


Microbial biodiversity in a Malaysian oil field and a systematic comparison with oil reservoirs worldwide.
Microbial diversity within formation water and oil from two compartments in Bokor oil reservoir from a Malaysian petroleum oil field was examined. A total of 1,056 16S rRNA gene clones were screened from each location by amplified ribosomal DNA restriction analysis. All samples were dominated by clones affiliated with Marinobacter, some novel Deferribacteraceae genera and various clones allied to the Methanococci. In addition, either Marinobacterium- or Pseudomonas-like operational taxonomic units were detected from either compartment. A systematic comparison with the existing pertinent studies was undertaken by analysing the microbial amplicons detected and the PCR primers used. The analyses demonstrated that bacterial communities were site specific, while Archaea co-occurred more frequently. Amplicons related to Marinobacter, Marinobacterium and Pseudomonas were detected in a number of the studies examined, suggesting they may be ubiquitous members in oil reservoirs. Further analysis of primers used in those studies suggested that most primer pairs had fairly broad but low matches across the bacterial and archaeal domains, while a minority had selective matches to certain taxa or low matches to all the microbial taxa tested. Thus, it indicated that primers may play an important role in determining which taxa would be detected.


The perineurial and blood-nerve barriers in experimental diabetes.
The permeabilities of the blood-nerve barrier and the perineurial barrier were investigated in alloxan and streptozotocin diabetic rats and in the mutant diabetic mouse [C57BL/Ks(db/db)]. In the mouse model both fluorescence and electron microscopic techniques were employed using Evans blue albumin (EBA) and horseradish peroxidase (HRP), respectively, as exogeneous tracers. In the rat models only HRP was used. Tracers were applied locally around the sciatic nerve in order to investigate the perineurial barrier, and systemically to detect changes in the blood nerve barrier. None of the models was found to show increased permeability across either of the barriers.


Skeletal muscle fiber composition and capillarization in patients with chronic heart failure: relation to exercise capacity and central hemodynamics.
Patients with chronic heart failure have structural and metabolic changes in skeletal muscle, which may be of importance for symptomatology. The origin of these changes are still unknown. The relationship between fiber composition and capillarization in skeletal muscle with exercise capacity and central hemodynamic variables was examined. Biopsies from the lateral vastus muscle were taken in 12 patients with chronic heart failure. Samples from eight normal subjects served as control samples. All patients underwent maximal exercise tests. Central hemodynamic variables were measured during exercise in five patients. The patients had a higher percentage of type II B fibers (P = .03) and fewer capillaries per fiber (P = .02) than the controls subjects. VO2 max correlated with the percentage of type I fibers, whereas the correlation with the type II A fibers was inverse. Cardiac index and pulmonary capillary wedge pressure at submaximal and maximal exercise were related to fiber type composition and relative fiber areas. Skeletal muscle fiber type composition and capillarization was changed in patients with chronic heart failure. These changes might influence exercise capacity. There were relationships between central hemodynamic variables and skeletal muscle changes. What the cause and effects were need further investigation.


Geographic disparities in state and district policies targeting youth obesity.
States and school districts nationwide have enacted policies targeting youth obesity, but many policies lack specific requirements or enforcement language. Geographic disparities in youth obesity could worsen if policies are weaker in areas with higher obesity prevalence. To determine if state or district policy strength varies with youth obesity prevalence across Census divisions. Policies in five domains related to nutrition and physical activity in schools were obtained from all states and nationally representative samples of 578 and 592 public school districts in the 2006-2007 and 2008-2009 school years, respectively. Policy language strength was rated on a 0-100 scale on both the state and district level. Regression models were used to determine if mean 2006-2007 strength scores, and changes in mean scores from 2006-2007 to 2008-2009, were associated with youth obesity prevalence across Census divisions. Analyses were conducted in 2010. State and district policies governing foods sold outside of school meal programs ("competitive foods") were stronger in 2006-2007 in the two divisions with the highest youth obesity prevalence (East South Central, West South Central). Furthermore, mean competitive food policy strength increased the most from 2006-2007 to 2008-2009 in these divisions. The West South Central had the weakest district physical education policies in 2006-2007, however, and was the only division in which average strength of district school meal policies decreased. State and districts in Census divisions with the highest youth obesity prevalence are taking steps to restrict competitive foods in schools, but many have not targeted other policy domains as aggressively.


Circulating Dickkopf-1 in hypoxic ischemic neonates.
The current study aimed to determine the serum level of Dickkopf-1 (Dkk-1) in peripheral blood of neonates with hypoxic ischemic encephalopathy (HIE). We measured serum levels of Dkk-1 by ELISA in neonates with HIE (n = 20) within 24 h from symptom onset and in healthy controls (n = 20). Dkk-1 serum levels increased significantly in HIE neonates than in healthy control. DKK-1 serum levels increased significantly in HIE neonates with convulsion, using multiple anti-convulsant drugs and those complicated with cranial ultrasound changes. Serum DKK-1 levels increased significantly in severe HIE patients. Our study provides for the first time the evidence of releasing Dkk-1 into the circulation of neonates with HIE with higher level in severe degree.


Mineralogy and fluid chemistry controls on lithium isotope fractionation during clay adsorption.
Our current understanding of controls on δ7Li variability and fractionation mechanisms is limited, complicating the interpretation of chemical weathering. The role of clay adsorption in Li isotope fractionation during chemical weathering has been confirmed. However, clay assemblage and fluid chemistry are not simple and often variable in weathering settings, potentially modulating Li isotope fractionation on Earth's surface. Here, this research investigated the patterns and processes of Li isotope fractionation during adsorption on kaolinite and smectite with fluid chemistry of 0.001 M NaCl, 0.5 M NaCl, and 0.001 M Na2HPO4. Specifically, the time-dependent experiments with the reaction period up to 15 days revealed that the steady state can be achieved within one day under neutral conditions. The concentration-dependent (initial Li concentration of 2 to 1000 μM) experiments confirmed the accumulation of Li+ in smectite interlayers and adsorption of Li+ only at the external surfaces of kaolinite. Using 0.5 M NaCl solution and the desorption experiments, we hypothesize that outer-sphere Li may exist in the interlayer sites, which can be replaced by excess Na+. In comparison, inner-sphere Li+ (unexchangeable) potentially dominates at the edge surface of clays. The presence of Na2HPO4 increases the binding capacity for Li+ adsorption, in particular for kaolinite. In all cases, 6Li is enriched on clay surfaces and interlayer spaces, consistent with field observations. Fluid chemistry may affect the degree of clay Li adsorption but exerted negligible impacts on isotope fractionation. For kaolinite, a wide variation (up to 30 ‰) in isotopic fractionation between adsorbed and aqueous Li (Δ7Liaq-ad) exists, conforming to a kinetic fractionation mechanism with a constant fractionation factor αad-aq of ~0.992. By contrast, the isotopic fractionation between Li adsorbed on smectite and Li+ left in solutions keeps constant (Δ7Liaq-ad of ~5 ‰), likely following an equilibrium isotope fractionation law with an αad-aq of ~0.995.


Modelling and optimization of NaOH-etched 3-D printed PCL for enhanced cellular attachment and growth with minimal loss of mechanical strength.
Despite having gained success in achieving intricate geometries for bone-graft fabrication, 3D printing technology still lacks good implant-tissue bonding. This can be addressed with alkaline surface post-treatment of 3D printed grafts, which improves the surface morphology and cellular response (attachment and proliferation), as shown in this study of polycaprolactone (PCL). The parameters for process optimization were NaOH-concentration, reaction temperature, and treatment time. Along with the hydrolysis reaction, its morphological implications at micro-level was also studied here for the first time. The modified surface was characterized by measuring surface porosity, surface roughness, and cellular response. A kinetic model was developed to correlate surface porosity with concentration, temperature and time. The concept of treatment intensity is introduced, which is a lumped parameter consisting of the product of the three governing parameters, which shows a concentration-temperature-time equivalency. With the increase in treatment intensity, surface porosity increased to ~60%, the surface roughness (RMS value) increased to ~700 nm, and cellular response improved till surface porosity reaches ~35%. This study establishes the importance of NaOH-PCL interaction and proposes that the surface reaction mechanism studied here can be exploited to enhance the in-vivo performance of bone grafts.


Molecular data on urinary glycoproteins with human leucocyte antigen (HLA) activity.
Two glycoproteins characterized by their serological activities (HLA-A9 and HLA-B12), their isoelectric points and their molecular weights were purified from urine from a patient suffering from tubular proteinuria (cystinosis). Their physicochemical properties as well as an important increase of their specific activities during the different purification steps suggested that they behave as human leucocyte antigens (HLA) which had been excreted into urine. Their amino acid compositions and N-terminal sequences were different to those described for HLA solubilized from cultured human lymphoblast cell lines. The N-terminal sequences of the two serologically active glycoproteins were identical to the N-terminal sequence of another recently purified human urinary glycoprotein called human complex-forming glycoprotein. The relationship between HLA, human complex-forming glycoprotein and the serologically active urinary glycoproteins is discussed.


[Exposition markers: mineralogical analysis in the sputum and the bronchoalveolar lavage--asbestos bodies--uncoated fibres].
The aim of mineralogical analysis of lung tissue, bronchoalveolar lavage (BAL) and sputum is to characterize individuals' exposure to asbestos fibres by identifying markers of this; asbestos bodies (AB) and uncoated fibres. The techniques of mineralogical analysis, habitually used to identify AB and uncoated fibres, are respectively optical microscopy (OM) and analytical electronic microscopy (EM). Correlations between levels of retention of AB in lung tissue, BAL and sputum have been established and validated threshold values indicating a high probability of significant exposure exist. These results must be interpreted in the context of clinical and occupational information. Mineralogical analysis is not suitable for use in routine medical screening but it can be considered when a source of exposure is not evident from the questionnaire since a positive analysis of BAL or of sputum is highly specific and thus useful to confirm an important retention of asbestos in the lung, which justifies medical follow-up. A negative result does not exclude previous significant asbestos exposure (frequent false negatives occur especially in sputum and biopersistence of chrysotile is lower than for amphiboles). Thus it can be a complementary tool for the assessment of asbestos exposure but its use imposes conditions for the collection and handling of samples.


Methods to study changing free amino acid pools during embryonic chick development.
During early embryogenesis of the chick, the yolk represents the only source of free amino acids. Leucine and glutamic acid, together with valine, predominate in this nonrenewable nutrient supply; taurine and phosphoethanolamine are present in the lowest concentration. The tyrosine level is already twice as high as phenylalanine. At embryo day 2 (E2), the not-yet fully functioning vitelline blood plexus demonstrates an amino acid profile that rather closely resembles that of the yolk, except for three amino acids. During the transfer process from yolk to plexus, a certain amount of glutamic acid appears to be metabolized to glutamine and aspartic acid. Taurine and phosphoethanolamine are highly concentrated in the circulation, with a ten-fold increase of taurine and a 50-fold rise in phosphoethanolamine. The tyrosine-to-phenylalanine ratio remains 2:1, but their levels relative to valine and other essential amino acids fall by roughly 50%. In the already contracting heart tube and the developing neural tissue on E2, taurine and phosphoethanolamine also accumulate preferentially; the levels in neural tissue are ten- and fivefold higher than in the circulation for the respective amino acids, whereas the heart has three time the plexus content of taurine and two times the amount of phosphoethanolamine. Tyrosine and phenylalanine, still in the same proportion, have risen to approximately twice the circulation values. The heart tube, unlike the primitive brain, concentrates leucine, whereas isoleucine accumulates in both organs. Injections of valine (50 mumol) and taurine (200 mumol) into the yolk on E1 demonstrate that a higher yolk content of an amino acid can result in a two- to threefold tissue increase.(ABSTRACT TRUNCATED AT 250 WORDS)


Single-dose methotrexate injection for treatment of ectopic pregnancy in women with relatively low levels of human chorionic gonadotropin.
To evaluate potential risk factors at admission for failed single-dose methotrexate injection in women with ectopic pregnancy. File review of a tertiary medical center in Israel yielded 102 consecutive patients with ectopic pregnancy initially treated with single-dose methotrexate. Overall successful treatment was defined as normalization of human chorionic gonadotropin (hCG) levels after the first or second injection. The mean pretreatment hCG level for the whole cohort was 2350 ± 2955 mIU/mL (median, 1187 mIU/mL; 90th percentile, 5000 mIU/mL). Resolution of ectopic pregnancy was achieved after a single injection in 76/102 (74.5%) women and after a second dose in 16/21 (76.2%) women. The remaining 10 (9.8%) patients were treated surgically after the first (5/76) or the second (5/26) dose. The overall success rate for methotrexate treatment was 90.2%. On multivariate stepwise logistic analysis, fetal cardiac activity was an independent predictor of a failed first methotrexate injection, and presence of a yolk sac and mass size were independent predictors of overall methotrexate failure and need for secondary surgery. In patients with ectopic pregnancy and relatively low levels of serum hCG, the most important predictors of overall failure of methotrexate injection were the presence of a yolk sac and mass size.


An immunoregulatory and tissue-residency program modulated by c-MAF in human TH17 cells.
Different types of effector and memory T lymphocytes are induced and maintained in protective or pathological immune responses. Here we characterized two human CD4+ TH17 helper cell subsets that, in the recently activated state, could be distinguished on the basis of their expression of the anti-inflammatory cytokine IL-10. IL-10+ TH17 cells upregulated a variety of genes encoding immunoregulatory molecules, as well as genes whose expression is characteristic of tissue-resident T cells. In contrast, IL-10- TH17 cells maintained a pro-inflammatory gene-expression profile and upregulated the expression of homing receptors that guide recirculation from tissues to blood. Expression of the transcription factor c-MAF was selectively upregulated in IL-10+ TH17 cells, and it was bound to a large set of enhancer-like regions and modulated the immunoregulatory and tissue-residency program. Our results identify c-MAF as a relevant factor that drives two highly divergent post-activation fates of human TH17 cells and provide a framework with which to investigate the role of these cells in physiology and immunopathology.


Inflammatory changes in affected muscles of facioscapulohumeral dystrophy.
A 34-year-old man developed slowly progressive muscular wasting of facioscapulohumeral (FSH) distribution with marked asymmetry. Clinical features, family history and laboratory findings were consistent with the diagnosis of FSH dystrophy. However, muscle biopsy revealed an inflammatory reaction with perivascular and endomysial infiltrate, myophagocytosis and invasion of the fibres by mononuclear cells. Monoclonal antibody analysis of the phenotypes of cells in the biopsy specimen suggested that cell-mediated immune response may occur during muscle fibre injury in FSH dystrophy.


Hematologic emergencies in the pediatric emergency room.
The complete blood count (CBC) describes the three hematopoietic lineages (i.e., the erythrocytes, leukocytes, and platelets), and it is an essential diagnostic component in numerous clinical situations. The pediatric CBC and hematologic problems in children may significantly differ from that of adults. In this article, special features of pediatric hematologic emergencies are highlighted.


The Montreal Cognitive Assessment is a valid cognitive screening tool for cardiac arrest survivors.
The survival rate of out-of-hospital cardiac arrest (OHCA) patients has increased over the past decades. This gives rise to a growing number of patients with hypoxic-ischemic brain damage and cognitive impairment. Currently, cognitive impairment is underdiagnosed in OHCA patients. There is a need for a validated cognitive screening instrument to identify patients with cognitive impairment. This study aimed to examine the diagnostic value of the Montreal Cognitive Assessment (MoCA) in patients after OHCA. Survivors (age ≥ 18 years) of OHCA completed the MoCA and a gold standard neuropsychological test battery, including tests for memory, attention, perception, language, reasoning, and executive functioning, at around one year after OHCA. Results of the MoCA are related to the results of the neuropsychological test battery. Analyses of diagnostic accuracy included receiver operating characteristics and calculation of predictive values. We included 54 OHCA survivors (mean age = 57.3, 74% male). The area under the curve (AUC) was 0.8, 95% CI [0.67, 0.93]. The MoCA showed excellent sensitivity of 86%, 95% CI [57, 98] and adequate specificity of 70.0%, 95% CI [53, 83] to detect cognitive impairment at the regular cut-off score of 26. The positive predictive value of the MoCA was 50%, 95% CI [30, 70] and the negative predictive value was 93%, 95% CI [76, 99]. This study shows that the MoCA may be a valid cognitive screening instrument for use in the OHCA patient population.


[Effect of cultivation for snail control on schistosomiasis in Changsha urban section of Xiangjiang River].
To study a reliable, friendly environmental and sustainable Oncomelania snail control method in marshlands of the Xiangjiang River. According to the special characteristics of the marshlands of the Xiangjiang River, smoothing and cultivating in beaches were applied for snail control. In sections with scarce distribution of snails, lowering parts of 4.4 beaches and raising other parts were added. Since 2003, the project has been practiced for 8 years. The average snail density declined by 99.83% with the simple smoothing beaches and then cultivation there. The average snail density declined by 100% with lowering parts of beaches and raising other parts and then cultivation. The schistosome infection rate was 0.31% with a declined rate of 93.84% in residents in 2006. The schistosome infection rate was 1.03% in 2010. The test result of water contamination was negative. The method of simple smoothing beaches and then cultivation can control the density of snails in marshlands and the method of lowering parts of beaches and raising other parts and then cultivation is more effective.


Arthroscopic Acetabular Labral Repair Using the Q-FIX Suture Anchor.
The goal of acetabular labral repair is to preserve/restore labral function. Maintaining labral function necessitates recreating the labrum's anatomy, especially avoiding a nonanatomic repair of the labrum to the acetabular rim. The purpose of this report is to detail the technique of acetabular labral repair using this Q-FIX all-suture anchor.


Inhibition by capsaicin of NADH-quinone oxidoreductases is correlated with the presence of energy-coupling site 1 in various organisms.
The NADH-ubiquinone reductase activity of the respiratory chains of several organisms was inhibited by capsaicin and dihydrocapsaicin, which are the pungent principles of red pepper. This inhibition was correlated with the presence of an energy transducing site in this segment of the respiratory chain. Where the NADH-quinone oxidoreductase segment involved an energy coupling site (e.g., in Paracoccus denitrificans, Escherichia coli, and Thermus thermophilus HB-8 membranes and bovine heart mitochondria), capsaicin acted as an inhibitor of ubiquinone reduction by NADH. In contrast, where this energy coupling site was absent (e.g., in Saccharomyces cerevisiae mitochondria and Bacillus subtilis membranes), there was no inhibition of NADH-ubiquinone reductase activity by capsaicin. The capsaicin inhibition of Paracoccus membranes was reversed by washing the membranes with medium containing bovine serum albumin. In the E. coli and Paracoccus membranes and bovine submitochondrial particles, capsaicin acted as a noncompetitive inhibitor for ubiquinone-1 at lower concentrations of ubiquinone-1 (less than 20 microM) and as a competitive inhibitor at higher concentrations of ubiquinone-1 (greater than 50 microM). In addition, the concentrations of capsaicin required for 50% inhibition of NADH oxidase activity of bovine submitochondrial particles were increased when ubiquinone-10 was added to the particles. The mechanism by which capsaicin inhibits the energy-transducing NADH-quinone oxidoreductase is discussed.


Preserving the Nipple Projection in Breast Reduction with the Free Nipple-Areolar Graft Technique: Purse-String Suture.
Breast reduction with the free nipple-areolar graft (FNG) technique has disadvantages such as loss of nipple projection, loss of nipple sensation, and depigmentation of the nipple-areolar complex. In this study, patients in whom a purse-string (PS) suture was used in the center of the de-epithelialized area to prevent loss of nipple projection were compared with patients who underwent the conventional method. A retrospective analysis of the patients who underwent breast reduction with the FNG was conducted in our department. Patients were divided into two groups according to the FNG placement. In the PS suture method group, a 1-cm-diameter circumferential suture was placed with a 5-0 Monocryl® (poliglecaprone 25) suture to gain a 6-mm-nipple projection. In the conventional method group, the FNG was placed directly over the de-epithelized area. Graft viability was evaluated after 3 weeks postoperatively. The final nipple projection and depigmentation were evaluated after 6 months postoperatively. The results were evaluated with statistical tests. The number of patients with the conventional method was 10, and the PS suture method was 12. There was no statistically significant difference between two groups regarding graft loss and depigmentation (p > 0.05). Nipple projection was significantly higher in the PS method group (p < 0.05). We observed that PS circumferential suture made an acceptable nipple projection compared to the conventional method in breast reduction with the FNG technique. Since the method is easy to apply and has relatively low risk, it would contribute to clinical practice. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .


Age-related change in task-evoked amygdala-prefrontal circuitry: A multiverse approach with an accelerated longitudinal cohort aged 4-22 years.
The amygdala and its connections with medial prefrontal cortex (mPFC) play central roles in the development of emotional processes. While several studies have suggested that this circuitry exhibits functional changes across the first two decades of life, findings have been mixed - perhaps resulting from differences in analytic choices across studies. Here we used multiverse analyses to examine the robustness of task-based amygdala-mPFC function findings to analytic choices within the context of an accelerated longitudinal design (4-22 years-old; N = 98; 183 scans; 1-3 scans/participant). Participants recruited from the greater Los Angeles area completed an event-related emotional face (fear, neutral) task. Parallel analyses varying in preprocessing and modeling choices found that age-related change estimates for amygdala reactivity were more robust than task-evoked amygdala-mPFC functional connectivity to varied analytical choices. Specification curves indicated evidence for age-related decreases in amygdala reactivity to faces, though within-participant changes in amygdala reactivity could not be differentiated from between-participant differences. In contrast, amygdala-mPFC functional connectivity results varied across methods much more, and evidence for age-related change in amygdala-mPFC connectivity was not consistent. Generalized psychophysiological interaction (gPPI) measurements of connectivity were especially sensitive to whether a deconvolution step was applied. Our findings demonstrate the importance of assessing the robustness of findings to analysis choices, although the age-related changes in our current work cannot be overinterpreted given low test-retest reliability. Together, these findings highlight both the challenges in estimating developmental change in longitudinal cohorts and the value of multiverse approaches in developmental neuroimaging for assessing robustness of results.


Can the Oral Bioavailability of the Discontinued Prostate Cancer Drug Galeterone Be Improved by Processing Method? KinetiSol® Outperforms Spray Drying in a Head-to-head Comparison.
Galeterone, a novel prostate cancer candidate treatment, was discontinued after a Phase III clinical trial due to lack of efficacy. Galeterone is weakly basic and exhibits low solubility in biorelevant media (i.e., ~ 2 µg/mL in fasted simulated intestinal fluid). It was formulated as a 50-50 (w/w) galeterone-hypromellose acetate succinate spray-dried dispersion to increase its bioavailability. Despite this increase, the bioavailability of this formulation may have been insufficient and contributed to its clinical failure. We hypothesized that reformulating galeterone as an amorphous solid dispersion by KinetiSol® compounding could increase its bioavailability. In this study, we examined the effects of composition and manufacturing technology (Kinetisol and spray drying) on the performance of galeterone amorphous solid dispersions. KinetiSol compounding was utilized to create galeterone amorphous solid dispersions containing the complexing agent hydroxypropyl-β-cyclodextrin or hypromellose acetate succinate with lower drug loads that both achieved a ~ 6 × increase in dissolution performance versus the 50-50 spray-dried dispersion. When compared to a spray-dried dispersion with an equivalent drug load, the KinetiSol amorphous solid dispersions formulations exhibited ~ 2 × exposure in an in vivo rat study. Acid-base surface energy analysis showed that the equivalent composition of the KinetiSol amorphous solid dispersion formulation better protected the weakly basic galeterone from premature dissolution in acidic media and thereby reduced precipitation, inhibited recrystallization, and extended the extent of supersaturation during transit into neutral intestinal media.


Reactivation performance of aerobic granules under different storage strategies.
Aerobic granules storage process is complicated and the affective mechanism is not very clear, which is influenced by storage temperature, storage substrate and storage time. The effects of storage temperature (-25 °C, 4 °C and room temperature) and storage substrate (distilled water and 400 mg/L glucose solution) on long-term storage and subsequently reactivation performance of aerobic granules were investigated in this study. The results showed that storage temperature had huge impact on the morphology and physical properties and storage substrate had relatively small influence on granules. Granules reactivation was a re-stability process of granules structure, physical properties and microbial communities. Storage at 4 °C was more suitable for maintenance of structural integrity and granules long-term stability storage. Granules stored at -25 °C achieved excellent settling ability after reactivation and PN/PS ratio was basically unchanged, which demonstrated that storage at -25°Cwas more suitable for the maintenance of the internal microstructure. Aerobic granules under different storage conditions could be reactivated after 10 days operation and the microbial activity (SOUR) could be fully restored. Furthermore granules stored at 4 °C obtained the best recovery performance, and granules at room temperature presented the worst restoration performance. In conclusion, no matter what the storage temperature or the storage substrate is, aerobic granules after long-term storage (8 months) could be restored within 10 days and stored granules could be successfully used as bioseed for reactor fast startup.


Development of an efficient gene targeting system in Colletotrichum higginsianum using a non-homologous end-joining mutant and Agrobacterium tumefaciens-mediated gene transfer.
The hemibiotrophic ascomycete Colletotrichum higginsianum is the casual agent of anthracnose disease of cruciferous plants. High efficiency transformation by Agrobacterium tumefaciens-mediated gene transfer has been established for this fungus. However, targeted gene mutagenesis through homologous recombination rarely occurs in C. higginsianum. We have identified and disrupted the C. higginsianum homologue of the human Ku70 gene, ChKU70, which encodes a protein that plays a role in non-homologous end-joining for repair of DNA breaks. chku70 mutants showed a dramatic increase in the frequency of integration of introduced exogenous DNA fragments by homologous recombination without any detectable phenotypic defects. This result demonstrates that the chku70 mutant is an efficient recipient for targeted gene mutagenesis in C. higginsianum. We have also developed a novel approach [named direct repeat recombination-mediated gene targeting (DRGT)] for targeted gene disruption through Agrobacterium tumefaciens-mediated gene transfer. DRGT utilizes homologous recombination between repeated sequences on the T-DNA flanking a partial fragment of the target gene. Our results suggest that DRGT in the chku70 mutant background could be a useful tool for rapid isolation of targeted gene disruptants in C. higginsianum.


A Digital Technique to Fabricate Segmental Individual Tooth Trays for Making an Impression of Multiple Crown Preparations.
A digital technique is described for designing and fabricating segmental individual tooth trays that can be used to make an impression of multiple preparations with high accuracy and efficiency. A digital model of prepared dentition was acquired. The segmental individual tooth trays with retention attachments and tissue stops were designed and 3D printed with light-polymerizing resin. The segmental impression of each section of the prepared dentition was made with the corresponding individual tooth tray loaded with the impression material. A final overall impression was made to pick up the segmental individual tooth trays with a full arch tray.


Older cancer patients and COVID-19 outbreak: Practical considerations and recommendations.
Since the COVID-19 outbreak started, it has been affecting mainly older individuals. Among the most vulnerable older individuals are those with cancer. Many published guidelines and consensus papers deal with prioritizing cancer care. Given the lack of high-quality evidence for management of cancer in older patients also in normal times, it is even more stringent to provide some resources on how to avoid both undertreatment and overtreatment in this population, who as of now is twice challenged to death, due to both a greater risk of getting infected with COVID-19 as well as from cancer not adequately addressed and treated. We hereby discuss some general recommendations (implement triage procedures; perform geriatric assessment; carefully assess comorbidity; promote early integration of palliative care in oncology; acknowledge the role of caregivers; maintain active take in charge to avoid feeling of abandonment; mandate seasonal flu vaccination) and discuss practical suggestions for specific disease settings (early-stage and advanced-stage disease for solid tumors, and hematological malignancies). The manuscript provides resources on how to avoid both undertreatment and overtreatment in older patients with cancer, who as of now is twice challenged to death, due to both a greater risk of getting infected with COVID-19 as well as from cancer not adequately addressed and treated.


Designing and evaluating assessment-based interventions to reduce stereotypy among adults with autism in a community job.
We designed and evaluated assessment-based interventions to reduce stereotypy among three adults with severe autism in a community job setting. An initial descriptive assessment, conducted during the regular work routine in the employing company, indicated that stereotypy occurred while the supported workers were waiting for work assignments or when they stopped working on an assigned task. An on-the-job functional analysis was then conducted; the results showed that the stereotypy of each worker was not maintained by socially-mediated consequences. Individualized interventions were then designed for increasing work behavior to compete with stereotypy by restructuring the job routine to reduce wait time and/or prompting and praising work behavior more frequently. The interventions increased work behavior and were accompanied by decreased stereotypy for each worker. Guidelines for practitioners and future research directions are offered, focusing on the use of assessment-based interventions for reducing stereotypy within community jobs.


Targeting the turnover of oncoproteins as a new avenue for therapeutics development in castration-resistant prostate cancer.
The current therapeutic armamentarium for castration-resistant prostate cancer (CRPC) includes second-generation agents such as the Androgen Receptor (AR) inhibitor enzalutamide and the androgen synthesis inhibitor abiraterone acetate, immunotherapies like sipuleucel-T, chemotherapies including docetaxel and cabazitaxel and the radiopharmaceutical radium 223 dichloride. However, relapse of CRPC resistant to these therapeutic modalities occur rapidly. The mechanisms of resistance to these treatments are complex, including specific mutations or alternative splicing of oncogenic proteins. An alternative approach to treating CRPC may be to target the turnover of these molecular drivers of CRPC. In this review, the mechanisms by which protein stability of several oncoproteins such as AR, ERG, GR, CYP17A1 and MYC, will be discussed, as well as how these findings could be translated into novel therapeutic agents.


A potential anticancer dihydropyrimidine derivative and its protein binding mechanism by multispectroscopic, molecular docking and molecular dynamic simulation along with its in-silico toxicity and metabolic profile.
Human serum albumin (HSA) is the core protein in the systemic circulation and has a fundamental role in transportation and distribution of ligands in-vivo. In this study, a newly synthesized and patented anticancer dihydropyrimidine derivative; 4-(4-ethoxyphenyl)-5-(3,4,5- trimethoxybenzoyl)-3,4-dihydropyrimidin-2(1H)-one (DHP) was evaluated for its binding to HSA. Ligand-HSA interaction is significant factor to attribute the toxicity or therapeutic potential to a ligand. Multi-spectroscopic studies combined with molecular modelling and molecular dynamic simulation (MDS) were conducted to understand the HSA-DHP binding mechanism. In-silico evaluation of DHP for its toxicity and metabolism was also conducted. Reduction in the binding constants was observed from 6.71 × 104 - 4.5 × 103 at increased temperatures which indicates moderate binding and the interaction was found to follow a static quenching mechanism. Further, Site I on HSA for DHP was established by competition with site specific markers and the results were supported by molecular docking. The stability of the HSA-DHP complex was established with MDS studies. Thermodynamics parameters revealed involvement of hydrogen bonding and van der Waals forces for HSA-DHP binding. An in-silico evaluation of DHP for its toxicity and metabolism provided that the synthesized compound was potentially safe and could be a promising candidate for further studies.


A boronate-modified molecularly imprinted polymer labeled with a SERS-tag for use in an antibody-free immunoassay for the carcinoembryonic antigen.
An antibody-free immunoassay that makes use of a boronate affinity molecularly imprinted polymer (MIP) and surface enhanced Raman scattering (SERS) tags is described. It was applied to the specific determination of the carcinoembryonic antigen (CEA) in human serum. For the preparation of the boronate affinity array, a polymer capable of adsorbing glycoproteins was first synthesized on the surface of a glass slide with four spots using 4-vinylbenzeneboronic acid (VPBA) as the functional monomer, ethylene glycol dimethacrylate (EGDMA) as the crosslinking agent, and ethylene glycol and cyclohexanol as porogens. The surface of the VPBA-Co-EGDMA can bind target glycoproteins. After specific capture of the glycoprotein, a "MIP-target glycoprotein-SERS tag" sandwich structure was formed by covalent interaction between the SERS nanotag (consisting of gold nanoparticles and 4-mercaptophenylboronic acid [MPBA]). CEA can be quantified in spiked serum with a detection limit of 0.1 ng·mL-1 via the specific Raman band at 1098 cm-1. Graphical abstractSchematic representation of the boronate affinity molecularly imprinted polymer (MIPs) array-based SERS sensor for rapid and sensitive detection of the carcinoembryonic antigen (CEA) from human serum. The boronate affinity MIPs array are used as capture probes, and MPBA@AuNPs are used as SERS tags.


Diverse Functions of Plasma PAF-AH in Tumorigenesis.
This chapter is focused on the role of the plasma form of platelet-activating factor-acetylhydrolase (PAF-AH), heretofore referred to as PAF-AH, in tumorigenic responses. Biochemical and other properties of this enzyme were discussed in detail in chapter "Plasma PAF-AH (PLA2G7): Biochemical Properties, Association with LDLs and HDLs, and Regulation of Expression" by Stafforini and in other chapters. Although phospholipases tend not to be drivers of tumorigenesis themselves, these enzymes and the lipid mediators whose levels they regulate interact with a variety of oncogenes and tumor suppressors [1]. Like other phospholipases, the functions of PAF-AH in cancer likely are related to its ability to regulate the levels of lipid mediators that participate in cellular processes related to initial tumorigenic events (e.g., proliferation, growth, inflammation) and/or spreading of the disease (e.g., matrix metalloproteinase secretion, actin cytoskeleton reorganization, migration, and angiogenesis) [1]. The importance of substrates and products of PAF-AH on key cellular functions has been evaluated in cell-based analyses which revealed that these metabolites can have pro- and antitumorigenic functions. Studies in genetically engineered mice lacking PAF-AH expression and genetic manipulation of PAF-AH levels in cancer cells demonstrated diverse functions of the protein in models of melanoma, prostate cancer, colon cancer, and others. The following sections highlight lessons learned from studies in cell lines and in mouse models regarding the diversity of functions of PAF-AH in cancer, and the potential of PAFAH transcripts, protein, and/or activity levels to become cancer biomarkers and therapeutic targets.


Balance and its Clinical Assessment in Older Adults - A Review.
Human beings rely on multiple systems to maintain their balance as they perform their activities of daily living. These systems may be undermined functionally by both disease and the normal aging process. Balance impairment is associated with increased fall risk. This paper examines the dynamic formulation of balance as activity and reviews the biological mechanisms for its control. A "minimal-technology" scheme for its clinical evaluation in the ambulatory care setting is proposed. The PubMed, Scopus and CINAHL databases were searched for relevant articles using the following terms in combination with balance: aging, impairment, control mechanisms, clinical assessment. Only articles which describe test procedures, their psychometrics and rely exclusively on equipment found in a regular physician office were reviewed. Human bipedal stance and gait are inherently low in stability. Accordingly, an elaborate sensory apparatus comprising visual, vestibular and proprioceptive elements, constantly monitors the position and movement of the body in its environment and sends signals to the central nervous system. The sensory inputs are processed and motor commands are generated. In response to efferent signals, the musculoskeletal system moves the body as is necessary to maintain or regain balance. The combination of senescent decline in organ function and the higher prevalence of diseases of the balance control systems in older adults predisposes this population subset to balance impairment. Older adults with balance impairment are likely to present with "dizziness". The history should concentrate on the first experience, with an attempt made to categorize it as a Drachman type. Since the symptomatology is often vague, several of the recommended physical tests are provocative maneuvers aimed at reproducing the patient's complaint. Well-validated questionnaires are available for evaluating the impact of "dizziness" on various domains of patient's lives, including their fear of falling. Aspects of a good history and physical examination not otherwise addressed to balance function, such as medications review and cognitive assessment, also yield information that contributes to a better understanding of the patient's complaint. Ordinal scales, which are aggregates of functional performance tests, enable detailed quantitative assessments of balance activity. The integrity of balance function is essential for activities of daily living efficacy. Its deterioration with aging and disease places older adults at increased risk of falls and dependency. Balance can be effectively evaluated in the ambulatory care setting, using a combination of scalar questionnaires, dedicated history-taking and physical tests that do not require sophisticated instrumentation.


Draft genome sequence of Clostridium sporogenes PA 3679, the common nontoxigenic surrogate for proteolytic Clostridium botulinum.
Clostridium sporogenes PA 3679 is widely used as a nontoxigenic surrogate for proteolytic strains of Clostridium botulinum in the derivation and validation of thermal processes in food. Here we report the draft assembly and annotation of the C. sporogenes PA 3679 genome. Preliminary analysis demonstrates a high degree of relatedness between C. sporogenes PA 3679 and sequenced strains of proteolytic C. botulinum.


[Renal side effects of long-term lithium therapy].
Lithium is widely used in the treatment of bipolar disorders. Long-term administration of lithium often leads to side effects concerning the subjects: nephrology, endocrinology and surgery. This review emphasizes nephrotoxicity.Lithium treatment may disturb responsiveness to antidiuretic hormone (ADH), causing a nephrogenic diabetes insipidus. Furthermore long-term lithium therapy may trigger hyperparathyreoidism with hypercalcemia and chronic interstitial nephritis with development of microcysts. Long-term patients have an increased risk to develop impaired renal function. Lithium-induced endstage renal disease is rare. Termination of lithium treatment may decrease the risk of progression.To ensure security of lithium treatment regular controls of urine osmolarity, lithium-, creatinine- , thyroid stimulating hormone- and calcium-levels are essential. Patients with decreased renal function should be referred to a specialist early.


Cloning and characterization of the genes for ribosomal proteins L10 and L12 from Synechocystis Sp. PCC 6803: comparison of gene clustering pattern and protein sequence homology between cyanobacteria and chloroplasts.
The endosymbiont theory proposes that chloroplasts have originated from ancestral cyanobacteria through a process of engulfment and subsequent symbiotic adaptation. The molecular data for testing this theory have mainly been the nucleotide sequence of rRNAs and of photosystem component genes. In order to provide additional data in this area, we have isolated genomic clones of Synechocystis DNA containing the ribosomal protein gene cluster rplJL. The nucleotide sequence of this cluster and flanking regions was determined and the derived amino acid sequences were compared to the available homologous sequences from other eubacteria and chloroplasts. In Escherichia coli these two genes are part of a larger cluster, i.e., rplKAJL-rpoBC. In Synechocystis, the genes for the RNA polymerase subunit (rpoBC) are shown to be widely separated from the r-protein genes. The Synechocystis gene arrangement is similar to that in the chloroplast system, where the rpoBC1C2 and rplKAJL clusters are separated and located in two cell compartments, the chloroplast and the nucleus, respectively.


Electrophysiological characterization of sodium-activated potassium channels in NG108-15 and NSC-34 motor neuron-like cells.
The electrical properties of Na(+) -activated K(+) current (I(K(Na)) ) and its contribution to spike firing has not been characterized in motor neurons. We evaluated how activation of voltage-gated K(+) current (I(K) ) at the cellular level could be coupled to Na(+) influx through voltage-gated Na(+) current (I(N) (a) ) in two motor neuron-like cells (NG108-15 and NSC-34 cells). Increasing stimulation frequency altered the amplitudes of both I(Na) and I(K) simultaneously. With changes in stimulation frequency, the kinetics of both I(Na) inactivation and I(K) activation were well correlated at the same cell. Addition of tetrodotoxin or ranolazine reduced the amplitudes of both I(Na) and I(K) simultaneously. Tefluthrin (Tef) increased the amplitudes of both I(Na) and I(K) throughout the voltages ranging from -30 to + 10 mV. In cell-attached recordings, single-channel conductance from a linear current-voltage relation was 94 ± 3 pS (n = 7). Tef (10 μm) enhanced channel activity with no change in single-channel conductance. Tef increased spike firing accompanied by enhanced facilitation of spike-frequency adaptation. Riluzole (10 μm) reversed Tef-stimulated activity of K(Na) channels. In motor neuron-like NSC-34 cells, increasing stimulation frequency altered the kinetics of both I(Na) and I(K) . Modelling studies of motor neurons were simulated to demonstrate that the magnitude of I(K(Na)) modulates AP firing. There is a direct association of Na(+) and K(Na) channels which can provide the rapid activation of K(Na) channels required to regulate AP firing occurring in motor neurons.


Color-coding and human factors engineering to improve patient safety characteristics of paper-based emergency department clinical documentation.
Investigators studied an emergency department (ED) physical chart system and identified inconsistent, small font labeling; a single-color scheme; and an absence of human factors engineering (HFE) cues. A case study and description of the methodology with which surrogate measures of chart-related patient safety were studied and subsequently used to reduce latent hazards are presented. Medical records present a challenge to patient safety in EDs. Application of HFE can improve specific aspects of existing medical chart organization systems as they pertain to patient safety in acute care environments. During 10 random audits over 5 consecutive days (573 data points), 56 (9.8%) chart binders (range 0.0-23%) were found to be either misplaced or improperly positioned relative to other chart binders; 12 (21%) were in the critical care area. HFE principles were applied to develop an experimental chart binder system with alternating color-based chart groupings, simple and prominent identifiers, and embedded visual cues. Post-intervention audits revealed significant reductions in chart binder location problems overall (p < 0.01), for Urgent Care A and B pods (6.4% to 1.2%; p < 0.05), Fast Track C pod (19.3% to 0.0%; p < 0.05) and Behavioral/Substance Abuse D pod (15.7% to 0.0%; p < 0.05) areas of the ED. The critical care room area did not display an improvement (11.4% to 13.2%; p = 0.40). Application of HFE methods may aid the development, assessment, and modification of acute care clinical environments through evidence-based design methodologies and contribute to safe patient care delivery.


[The action of pyruvate kinase and phosphoenolpyruvate on the development of experimental thrombosis].
Tests set up on rats and rabbits showed the ability of phospho-enolpyruvate and pyruvate-kinase to lower the aggregation of thrombocytes and prevent formation of experimental microthrombosis. The joint introduction of these compounds in most experiments resulted in potentiation of the antithrombotic effect.


Genome-wide identification and characterization of the C2 domain family in Sorghum bicolor (L.) and expression profiles in response to saline-alkali stress.
The C2 domain family proteins in plants has been recently shown to be involved in the response to abiotic stress such as salt and drought stress. However, less information on C2 domain family members has been reported in Sorghum bicolor (L.), which is a tolerant cereal crop. To elaborate the mechanism of C2 domain family members in response to abiotic stress, bioinformatic methods were used to analyze this family. The results indicated that 69 C2 domain genes belonging to 5 different groups were first identified within the sorghum genome, and each group possessed various gene structures and conserved functional domains. Second, those C2 family genes were localized on 10 chromosomes 3 tandem repeat genes and 1 pair of repeat gene fragments were detected. The family members further presented a variety of stress responsive cis-elements. Third, in addition to being the major integral component of the membrane, sorghum C2 domain family proteins mainly played roles in response to abiotic and biotic stress with their organic transport and catalytic activity by specific location in the cell on the basis of gene ontology analysis. C2 family genes were differentially expressed in root, shoot or leaf, and shown different expression profiling after saline-alkali stress, which indicated that C2 family members played an important role in response to saline-alkali stress based on the transcription profiles of RNA-seq data and expression analysis by quantitative real-time polymerase chain reaction. Besides, most C2 family members were mainly located in cytoplasmi and nucleus. Weighted gene co-expression network analysis revealed three modules (turquoise, dark magenta and pink) that were associated with stress resistance, respectively. Therefore, the present research provides comprehensive information for further analysis of the molecular function of C2 domain family genes in sorghum. The online version contains supplementary material available at 10.1007/s12298-022-01222-3.


Low density lipoprotein mimics insulin action on autophagy and glucose uptake in endothelial cells.
Elevated plasma low density lipoprotein (LDL) is an established risk factor for cardiovascular disease. In addition to being able to cross the endothelial barrier to become accumulated in subendothelial space and thereby initiate atherosclerosis, LDL may exert a direct effect on vascular endothelial cells through activation of LDL receptor and its downstream signaling. Whether LDL can modulate the signaling for autophagy in endothelial cells is not clear. The present study firstly demonstrated that LDL can suppress endothelial autophagy through activation of the PI3K/Akt/mTOR signaling pathway and can promote glucose uptake by translocating glucose transporter 1 (GLUT1) from cytoplasm to cell membrane, actions similar to those of insulin. A co-immunoprecipitation assay found that LDL receptor (LDLR) and insulin receptor (IR) formed a complex in HUVECs. Knock down of the insulin receptor by small interfering RNA blocked the suppression of autophagy by LDL, as well as the signaling pathway involved. We conclude that LDL may mimic the action of insulin in endothelial cells, which might partly explain the increased incidence of diabetes in patients receiving some LDL-lowering therapy.


Oxidation of a methionine residue in subtilisin-type proteinases by the hydrogen peroxide/borate system--an active site-directed reaction.
Subtilisin-type proteinases (thermitase, subtilisin Carlsberg, alkaline proteinase ZIMET 10911, proteinase K) are partially inactivated by hydrogen peroxide in the alkaline pH range only in the presence of boric acid or phenylboronic acid. A model is presented to describe the inactivation mechanism. Both boric acid and perboric acid existing in equilibrium in the presence of hydrogen peroxide bind competitively at the active site of the enzyme. The inactivation, which is known to be caused by sulfoxide formation from the methionine residue in the active site (Stauffer, C.E. and Etson, D. (1969) J. Biol. Chem. 244, 5333-5338), is due to the enzyme-bound perboric acid species. The dissociation constants for the boric acid-thermitase and perboric acid-thermitase complexes are 36 +/- 7 and 4 +/- 1 mM, respectively. The first-order rate constant of inactivation is k = 0.63 +/- 0.14 min-1. The same mechanism of inactivation holds true for phenylboronic acid in alkaline hydrogen peroxide solutions.


Social isolation and subsequent health and well-being in older adults: A longitudinal outcome-wide analysis.
Social isolation has become a serious public health issue. However, most previous studies examine the relationship between social isolation and a single outcome. We aimed to conduct holistic assessments to understand the multidimensional impacts of social isolation on health and well-being. We used the three-wave data (2013, 2016, and 2019) obtained from the Japan Gerontological Evaluation Study. Our exposure, obtained from the 2016 survey, was the Social Isolation Index (SII) comprising five components: no partner, poor interaction with children, poor interaction with relatives, poor interaction with friends, and no social participation). We assessed 36 health and well-being outcomes across six dimensions obtained from the 2019 survey: physical/cognitive health, health behaviours, mental health, subjective well-being, social isolation, and cognitive social capital. Pre-exposure characteristics and prior outcome levels in 2013 were adjusted. We included 47,318 respondents for 4 outcomes (death, dementia, and functional disability) and 34,187 respondents for 32 other outcomes. The Bonferroni correction was used to correct for multiple tests. The total SII scores were associated with a wide range of health and well-being outcomes across the six dimensions. Specifically, we found a robust association between an SII score of four or greater with mortality (Odds ratio: 1.89; 95% CI: 1.46-2.43). Among the five components of the SII, poor interaction with friends and no social participation showed robust associations with a wide range of health and well-being. We also found some robust evidence regarding effect modification by gender and age in the associations between the components of the SII and health and well-being. Social isolation, specifically social interaction with friends and social participation, may affect a wide range of health and well-being among older adults.


Secondhand tobacco smoke exposure is associated with prolactin but not thyroid stimulating hormone among nonsmoking women seeking in vitro fertilization.
Prolactin (PRL) and thyroid stimulating hormone (TSH) serve important roles in the reproductive and other systems. Active smoking is associated with changes in PRL and TSH secretion, but the relationship between secondhand tobacco smoke (STS) exposure and these hormones is unclear. We measured PRL and TSH in serum as well as cotinine in follicular fluid (to estimate STS exposure) among 314 nonsmoking women undergoing in vitro fertilization treatment. We observed a significant increase in PRL concentrations (p=0.03) among STS-exposed nonsmokers compared to unexposed nonsmokers. There was no significant difference in TSH concentration (p>0.4) among those exposed to STS compared to those who were unexposed. STS exposure is associated with an increase in circulating PRL but not TSH levels. Future studies are needed to confirm our results, identify biological mechanisms involved, and better understand the potential clinical and public health implications.


Inactivation of human tumor cell pro-urokinase by granulocyte elastase.
Supernatant obtained from granulocytes stimulated in the presence of cytochalasin B by the chemotactic peptide N-formyl-norleucyl-leucyl-phenylalanyl-norleucyl-tyrosyl- lysine displayed an inhibitory effect on the plasmin-dependent conversion of tumor urokinase-type plasminogen activator proenzyme (pro-uPA) to the active form of uPA. Moreover, the supernatant was also found to inhibit the fibrinolytic activity of human vulva (A431) and breast (MCF7) carcinoma cell lines, which contain large amounts of pro-uPA, by 87% and 96%, respectively. By using eglin C (elastase inhibitor) and a monoclonal antibody to elastase (proteolytic activity blocker of the enzyme), elastase was identified as the key enzyme of the supernatant in these phenomena. Purified elastase converted pro-uPA to an enzymatically inactive molecule composed of two polypeptide chains of Mr = 33,000 and 22,000 linked to each other by a disulfide bond. Elastase-containing granulocytes were identified by immunohistochemistry techniques in the tissues of squamous cell carcinoma and adenocarcinoma of uterus. The cells were found close to the tumor cells and in the stroma surrounding the tumor nests. By immunohistochemical staining, uPA was also found in the tumor cells. Evidently, elastase released by chemotactically activated granulocytes, which are attracted into tumor tissues, may inhibit the conversion of pro-uPA to enzymatically active uPA in the tumor cells.


A prospective, randomized, double blind, placebo-controlled evaluation of the effects of eicosapentaenoic acid and docosahexaenoic acid on the clinical signs and erythrocyte membrane polyunsaturated fatty acid concentrations in dogs with osteoarthritis.
Osteoarthritis (OA) in dogs is a prevalent and serious condition. The most common treatment for the clinical signs of OA in dogs is the administration of nonsteroidal antiiflammatory pharmaceuticals. Omega-3 (n-3) fatty acids have been shown to reduce the clinical signs of osteoarthritis in dogs. The primary goals of this study were 1) to determine the effects of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on the clinical signs of OA in dogs, 2) to evaluate the effects of supplementation on the arachadonic acid (ARA)/ (EPA+DHA) algorithm and 3) to correlate alterations in the ARA/(EPA+DHA) with changes in the clinical signs of canine OA. Seventy-eight client owned dogs were enrolled in a prospective, randomized, double-blind, placebo controlled clinical trial. Dogs were randomized to placebo oil or triglyceride n-3 oil (providing an average dose of 69mg EPA+DHA/kg/day). Orthopedic examinations and blood analyses were performed at baseline, day 42, and day 84. A single investigator confirmed a diagnosis of OA of the coxofemoral joints and/or stifle joints in all dogs. Seventy-four dogs completed the trial. All clinical outcomes for measuring discomfort, lameness, and joint severity at day 84 and all blood metrics at day 42 and day 84 significantly (p<0.05) improved compared with placebo. No major side effects were observed. This study demonstrated that the daily supplementation of a dogs diet with EPA and DHA shifts the blood fatty acid concentrations correlating to relief of clinical signs associated with OA in dogs.


Rapid and steady-state amino acid transport in perfused human fibroblasts and colon adenocarcinoma cells: effects of methotrexate.
Initial and steady-state uptakes of serine and phenylalanine by human fibroblasts and human colon tumour cells were studied applying a double isotope dilution technique to perfused populations of cultivated cells retained on microcarrier beads. This new method permits the differentiation of the unidirectional transport parameters and can also distinguish between membrane-associated processes and independently intracellular events in isolated cells. High initial L-serine uptake values in colon adenocarcinoma cells became negative under steady-state conditions. To determine if the observed negative L-serine uptake was produced by the rapid efflux of intracellular L-[3H]serine, the cells were treated with methotrexate (MTX) (an inhibitor of cytosolic dihydrofolate reductase). The modified curve of L-[3H]serine uptake after MTX treatment suggests that, under these experimental conditions, net serine transport is non concentrative in colon tumour cells and could be modulated by the rate of intracellular serine metabolism; it also suggests that MTX does not directly affect serine transport in perfused human colon adenocarcinoma cells. Initial and steady-state uptakes of phenylalanine were high in both fibroblasts and tumour cells and were unaffected by MTX treatment.


Prognostic Factors of MS Conversion in Optic Neuritis.
Investigators from Children's Hospital Aschaffenburg, Germany; University of Manchester, Manchester, United Kingdom; University Children's Hospital Tübingen, Tübingen, Germany; and other international centers studied prognostic factors in optic neuritis.


Efficacy of itraconazole in treating AIDS-associated infections due to Candida krusei.
Candida krusei is an emerging pathogen, both in HIV negative immunocompromised patients and in HIV seropositive patients. Its onset appears to be due, at least in part, to the use of fluconazole. In HIV positive patients, a long term prophylactic use of fluconazole may select some less pathogenic Candida species, as C. krusei, that may determine persistent oral candidiasis and emerge as systemic pathogen. Itraconazole appears efficacious in treating AIDS-associated C. krusei infections.


Superficial reticular degeneration of Koby.
Koby in 1927 described a progressive, superficial corneal degeneration in a middle-aged woman that led to a painless loss of vision. This condition was characterized by the appearance of a central, fine, white reticulum at the level of Bowman's layer. The epithelium was thickened and bedewed, giving a faint brownish opalescence and providing a striking contrast to the white reticulum. We evaluated a patient with this rare corneal degeneration, and present clinical findings as well as the results of a superficial corneal biopsy. The biopsy was evaluated by light and electron microscopy and provides the initial documentation of this entity as an atypical form of band keratopathy.


Euthymic and depressed bipolar patients are characterized by different RNA editing patterns in blood.
Bipolar disorder (BD) is a worldwide leading cause of disability. Inflammation roles in this disease is well established. ADAR1-mediated RNA editing is one of the key mechanisms regulating the inflammatory response. We have identified a panel of RNA editing-based blood biomarkers which allowed to discriminate unipolar from BD depression with high accuracy. We confirmed here the diagnostic value of this panel in a new cohort of BD patients recruited in Brazil. We also identified new combinations which allow a clear discrimination of BD from healthy controls and among BD subgroups, confirming that RNA editing is a key mechanism in BD.


Cranial floor fracture: A growing orbital roof fracture with encephalocele - Case Report.
Orbital roof fractures are among the rarest of craniofacial fractures. The mechanism of injury is typically a high-impact blunt force vector directly to the orbit or forehead. Most patients are males between 20 and 40 years old, involved in motor vehicle accidents. Although most orbital roof fractures are managed conservatively, there is a significant risk of ophthalmologic and neurologic complications. Detailed craniofacial examination and high-resolution CT imaging is necessary for diagnosis. A multidisciplinary team approach is required for these challenging fractures.


Systematic literature review informing the 2018 update of the EULAR recommendation for the management of large vessel vasculitis: focus on giant cell arteritis.
To analyse the current evidence for the management of large vessel vasculitis (LVV) to inform the 2018 update of the EULAR recommendations. Two systematic literature reviews (SLRs) dealing with diagnosis/monitoring and treatment strategies for LVV, respectively, were performed. Medline, Embase and Cochrane databases were searched from inception to 31 December 2017. Evidence on imaging was excluded as recently published in dedicated EULAR recommendations. This paper focuses on the data relevant to giant cell arteritis (GCA). We identified 287 eligible articles (122 studies focused on diagnosis/monitoring, 165 on treatment). The implementation of a fast-track approach to diagnosis significantly lowers the risk of permanent visual loss compared with historical cohorts (level of evidence, LoE 2b). Reliable diagnostic or prognostic biomarkers for GCA are still not available (LoE 3b).The SLR confirms the efficacy of prompt initiation of glucocorticoids (GC). There is no high-quality evidence on the most appropriate starting dose, route of administration, tapering and duration of GC (LoE 4). Patients with GCA are at increased risk of dose-dependent GC-related adverse events (LoE 3b). The addition of methotrexate or tocilizumab reduces relapse rates and GC requirements (LoE 1b). There is no consistent evidence that initiating antiplatelet agents at diagnosis would prevent future ischaemic events (LoE 2a). There is little evidence to guide monitoring of patients with GCA. Results from two SLRs identified novel evidence on the management of GCA to guide the 2018 update of the EULAR recommendations on the management of LVV.