The interaction of cytochrome c with monolayers of phosphatidylethanolamine.
1. The interaction between [(14)C]carboxymethylated cytochrome c and monolayers of egg phosphatidylethanolamine at the air/water interface has been investigated by measurements of surface radioactivity, pressure and potential. 2. On adding (14)C-labelled cytochrome c to the subphase under monolayers with a surface pressure below 24dynes/cm. there was an initial surface pressure increment as the protein penetrated, followed by an adsorption that could be detected only by a continued increase in the surface radioactivity. 3. Above film pressures of 24dynes/cm. only adsorption was observed, i.e. an increment in surface radioactivity with none in surface pressure. 4. The changes in surface parameters with penetration of cytochrome c added to the subphase were indirectly proportional to the initial pressure of the monolayer. With hydrogenated phosphatidylethanolamine the constant of proportionality was increased but penetration again ceased at 24dynes/cm. 5. On compressing a phosphatidylethanolamine film containing penetrated cytochrome c to 40dynes/cm. only a proportion of the protein was ejected on a subphase of 10mm-sodium chloride, whereas on a subphase of m-sodium chloride nearly all the protein was lost. 6. With both penetration and adsorption only a small proportion of the added cytochrome c interacted with the phospholipid films, and initially the amount bound was proportional to the added protein concentration. There was no evidence of a stoicheiometric relationship between the protein and phospholipid or the build-up of multilayers. The bonded protein was not released by removing cytochrome c from the subphase. 7. The addition of m-sodium chloride to the subphase delays the rate of protein penetration into low-pressure films, but the final surface-pressure increment is not appreciably decreased. In contrast, m-sodium chloride almost completely stops adsorption on to films at all pressures. 8. When sodium chloride is added to the subphase below cytochrome c adsorbed to monolayers at high pressures, so that the final concentration is 1m, only a proportion of the protein is desorbed and this decreases as the time of the interaction increases. This indicates that adsorption is initially electrostatic, followed by the formation of non-ionic bonds. 9. Alteration of the subphase pH under a high-pressure film leads to a steady increase in adsorption from pH3 to 8.5 followed by a rapid fall to zero adsorption at pH11. 10. The penetration into phospholipid monolayers at 10dynes/cm. shows a rate that is consistent with the relative electrostatic status of the two components of the interaction as the subphase pH is varied between 3 and 10.5. The final equilibrium penetration shows a pronounced peak in the increments of surface pressure at pH9.0 although a similar peak is not observed in the surface radioactivity. This indicates that more residues of the protein are penetrating into the film at about this pH. 11. Determinations were made of the electrophoretic mobilities of phosphatidylethanolamine particles both alone and after interaction with cytochrome c. 12. The electrophoretic mobilities of cytochrome c adsorbed on lipid particles showed an isoelectric point below that of cytochrome c. This and the observations on the monolayers suggest that, with cytochrome c, protein-protein interactions are weak compared with other proteins.


In the eyes of the beholder: A non-self-report measure of workplace deviance.
Because employees may be reluctant to admit to performing deviant acts, the authors of this study reexamined the commonly used self-report measure of workplace deviance developed by R. J. Bennett and S. L. Robinson (2000). Specifically, the self-report measure was modified into a non-self-report measure based on multiple other-reported assessments to address methodological concerns with self-reported information regarding deviant workplace behaviors. The authors assessed the psychometric properties of this new measure by first conducting an exploratory factor analysis, which indicated a 3-factor structure (production deviance, property deviance, and personal aggression). Subsequent confirmatory factor analysis on a different sample verified these findings. Taken together, the results suggest that the content and psychometric qualities of this non-self-report measure of workplace deviance closely represent S. L. Robinson and R. J. Bennett's (1995) original typology of workplace deviance. The potential usefulness of this measure in organizational studies is discussed. (PsycINFO Database Record (c) 2009 APA, all rights reserved).


Linaclotide: a novel agent for chronic constipation and irritable bowel syndrome.
The pharmacology, pharmaco-kinetics, and clinical efficacy and safety of linaclotide in the management of chronic constipation (CC) and constipation-predominant irritable bowel syndrome (IBS-C) are reviewed. Linaclotide (Linzess, Forest Pharmaceuticals) is a 14-amino acid peptide indicated for the treatment of adults with CC and IBS-C. Linaclotide acts on guanylate cyclase-C receptors on the luminal membrane to increase chloride and bicarbonate secretions into the intestine and inhibit the absorption of sodium ions, thus increasing the secretion of water into the lumen and improving defecation; the drug is minimally absorbed into the systemic circulation. Linaclotide is approved by the Food and Drug Administration (FDA) for oral once-daily administration at doses of 145 μg for CC and 290 μg for IBS-C. In placebo-controlled Phase III clinical trials, linaclotide significantly increased weekly spontaneous bowel movements and complete spontaneous bowel movements (CSBMs) while reducing abdominal pain in patients with CC. In patients with IBS-C, linaclotide was demonstrated to be effective in meeting FDA-recommended endpoints such as reductions of at least 30% from baseline in abdominal pain scores and CSBM frequency. The most common adverse effect of linaclotide is diarrhea, which was reported in 16-20% of clinical trial participants. Linaclotide is an important advance in the treatment of CC and IBS-C, with a novel mechanism of action resulting in accelerated intestinal transit. In clinical trials, linaclotide demonstrated efficacy relative to placebo for treatment of both CC and IBS-C. Linaclotide's adverse effects are generally mild and confined to the gastrointestinal tract.


Influence of myoglobin on the antibacterial activity of carvacrol and the binding mechanism between the two compounds.
Myoglobin (MB), a pigmentation protein, can adversely affect the antibacterial activity of carvacrol (CAR) and weaken its bacteriostasis effect. This study aimed to clarify the influence of MB on the antibacterial activity of CAR and ascertain the mechanism involved in the observed influence, especially the interaction between the two compounds. Microbiological analysis indicated that the presence of MB significantly suppressed the antibacterial activity of CAR against Listeria monocytogenes. Ultraviolet-visible spectrometry and fluorescence spectroscopic analysis confirmed the interaction between CAR and MB. The stoichiometric number was determined as ~0.7 via double logarithmic Stern-Volmer equation analysis, while thermodynamic analysis showed that the conjugation of the two compounds occurred as an exothermal reaction (ΔH° = -32.3 ± 11.4 kJ mol-1 and ΔS° = -75 J mol-1 K-1 ). Circular dichroism, Fourier transform infrared spectroscopy and nuclear magnetic resonance spectroscopy showed hydrogen bonding in the carvacrol-myoglobin complex (CAR-MB). Molecular docking analysis confirmed that amino acid residues, including GLY80 and HIS82, were most likely to form hydrogen bonds with CAR, while hydrogen bonds represented the main driving force for CAR-MB formation. CAR antibacterial activity was significantly inhibited by the presence of MB in the environment due to the notable reduction in the effective concentration of CAR caused by CAR-MB formation. © 2023 Society of Chemical Industry.


Effects of 17beta-estradiol and testosterone on hepatic mRNA/protein levels and catalytic activities of CYP2M1, CYP2K1, and CYP3A27 in rainbow trout (Oncorhynchus mykiss).
There is growing concern that exposure to chemicals in the environment can disrupt the endocrine systems of wildlife and humans, causing reproductive problems or other adverse effects. The expression of many cytochrome P450s (CYPs) is under hormonal control, hence, levels of these enzymes can be affected by exposure to endocrine-disrupting chemicals. Previous research has reported that treatment of fish and other animals with the estrogenic and androgenic hormones 17beta-estradiol (E2) and testosterone (T) alters the P450 content or enzyme activities in the treated animals. However, the results of many of these studies are either incomplete or in disagreement and in most cases the effect on specific P450 forms has not been determined. Therefore, to better understand the effects of gonadal hormones on the expression of P450s and their associated enzyme activities, it was of interest to undertake a comprehensive investigation of the transcriptional and translational expression of three constitutive hepatic P450s in the rainbow trout (Oncorhynchus mykiss) following hormone exposure. Accordingly, juvenile trout were injected intraperitoneally with propylene glycol vehicle and the most active estrogenic and androgenic hormones E2 (3 mg/kg) or T (3 mg/kg) on days 1, 4, 7, 13, and 15 and euthanized on day 19. After treatment with E2, hepatic microsomes showed significantly lower levels (percentage of control) in total P450 contents (52%), lauric acid hydroxylase (32%), and 6beta-progesterone hydroxylase activities (27%), [(3)H]aflatoxin-DNA binding (31%), and the protein levels of individual cytochrome P450s (CYPs) LMC1 (CYP2M1), LMC2, (CYP2K1), and LMC5 (CYP3A27) (average for three isoforms a reduction to 29% of control values) with only minor differences between sexes. Treatment with T had either no effect or resulted in small increases in total P450 in males (42%), in lauric acid hydroxylase in females (24%), and in 6beta-progesterone hydroxylase activity in males (21%). Biological variabilities among fish were high and a polymorphic or new LMC2-like form was detected at about 52 kDa in some liver microsomal samples after exposure of fish to either hormone. Female liver RNAs were analyzed through Northern blots and an average decrease of 94% in CYP2 M1, CYP2K1, and CYP3A27 mRNA levels occurred in the E2-treated trout. In livers from T-treated trout, the changes of mRNA levels of CYP2M1 and CYP3A27 were negligible, but CYP2K1 mRNA level decreased by about 60%. Additional CYP2K1 cDNA hybridizable mRNAs were seen in some fish as faint bands at about 2.8 kb for both hormone treatments. Results of this study, therefore, indicated that E2 down-regulated while T produced small but variable effects on the hepatic mRNA/protein levels of CYP2K1, CYP2M1, and CYP3A27 in juvenile rainbow trout. This study, therefore, suggests that exposure of fish and other wildlife to environmental endocrine disruptors, especially estrogen mimics, can adversely affect a number of physiological processes through mechanisms involving altered levels of expression of specific P450 isozymes.


Gastric digestion modifies absorption of butterfat into lymph chylomicrons in rats.
Our objective was to characterize the time course of mesenteric lymph output, lipid composition and size of lymph chylomicrons in rats given gastric infusion of lipid emulsions containing defined fractions of butterfat, palm oil or corn oil. The concentrations of cholesterol, triacylglycerol (TAG) and phospholipid in lymph obtained before lipid infusion were 1.4-2.5-fold greater in rats chronically fed palm oil or solid butterfat compared with corn oil or liquid butterfat (P = 0.02). Total lymph chylomicron TAG output (mg/24 h) stimulated by gastric lipid infusion was 21% greater with corn oil compared with all saturated fats (P = 0.02). Total lymph chylomicron cholesterol output was 1.3-8.6-fold greater than the amount infused in all groups (P = 0.03) and was independent of the amount of cholesterol infused. The size distribution as well as the mean, median and modal diameters of lymph chylomicrons isolated during peak lymphatic TAG output were not significantly different among treatments. The fatty acid and TAG profiles of lymph chylomicrons obtained from rats infused with corn or palm oil did not differ significantly from that of the emulsion infused. In contrast, gastric lipolysis of butterfat significantly modified the lipid composition of lymph chylomicrons. We observed progressive disappearance of short- and medium-chain fatty acids in gastric contents and an absence of detectable short-chain fatty acids with concurrent proportionate increases in long-chain fatty acids and large TAG molecules in lymph chylomicrons compared with butterfat emulsions. These studies demonstrate that gastric digestion is an important modifier of lipid absorption.


Chromosomal location of a Triticum timopheevii--derived powdery mildew resistance gene transferred to common wheat.
A dominant powdery mildew resistance gene introduced from Triticum timopheevii in line 146-155-T of common wheat, Triticum aestivum, was located on chromosome 6B by monosomic analysis. Restriction fragment length polymorphism (RFLP) and microsatellite analyses detected the presence of a T. timopheevii segment, translocated to chromosome 6B, with breakpoints between the loci Xpsr8/Xpsr964 on 6BS and Xpsr154/Xpsr546 on 6BL. The novel powdery mildew resistance gene, which has been designated Pm27, was shown to cosegregate with the microsatellite locus Xpsp3131, which is located on the introgressed T. timopheevii segment. The molecular data confirm the location of Pm27 on the translocated 6B chromosome.


MRL proteins cooperate with activated Ras in glia to drive distinct oncogenic outcomes.
The Mig10/RIAM/Lpd (MRL) adapter protein Lpd regulates actin dynamics through interactions with Scar/WAVE and Ena/VASP proteins to promote the formation of cellular protrusions and to stimulate invasive migration. However, the ability of MRL proteins to interact with multiple actin regulators and to promote serum response factor (SRF) signalling has raised the question of whether MRL proteins employ alternative downstream mechanisms to drive oncogenic processes in a context-dependent manner. Here, using a Drosophila model, we show that overexpression of either human Lpd or its Drosophila orthologue Pico can promote growth and invasion of RasV12-induced cell tumours in the brain. Notably, effects were restricted to two populations of Repo-positive glial cells: an invasive population, characterized by JNK-dependent elevation of Mmp1 expression, and a hyperproliferative population lacking elevated JNK signalling. JNK activation was not triggered by reactive immune cell signalling, implicating the involvement of an intrinsic stress response. The ability to promote dissemination of RasV12-induced tumours was shared by a subset of actin regulators, including, most prominently, Chicadee/Profilin, which directly interacts with Pico, and, Mal, a cofactor for serum response factor that responds to changes in G:F actin dynamics. Suppression of Mal activity partially abrogated the ability of pico to promote invasion of RasV12 tumours. Furthermore, we found that larval glia are enriched for serum response factor expression, explaining the apparent sensitivity of glial cells to Pico/RasV12 overexpression. Taken together, our findings indicate that MRL proteins cooperate with oncogenic Ras to promote formation of glial tumours, and that, in this context, Mal/serum response factor activation is rate-limiting for tumour dissemination.


Human experience in space.
Dr. Carr recounts his own experience in space on the Skylab III mission. He notes that medical research in space continues today, and compares current Shuttle missions with those of Skylab.


Personalized Feedback on Symptom Dynamics of Psychopathology: A Proof-of-Principle Study.
In the proposed symptom network approach to psychopathology, psychiatric disorders are assumed to result from the (causal) interplay between symptoms. By implementing this approach we explored whether individual feedback on symptom dynamics complements current categorical classification and treatment. The aim of this proof-of-principle case-study was to explore the feasibility, acceptability and usability of this transdiagnostic approach. A female patient, aged 67, suffering from treatment resistant anxious and depressive symptoms was treated in our tertiary outpatient clinic for old age psychiatry. She participated in ecological momentary assessments (EMA), which involved intensive repeated measurements of mood and context-related items during two weeks. Visualizations of the interplay between the items were provided by network graphs and were discussed with the patient. Network graphs were discussed with the patient. For example, it was hypothesized and discussed with the patient that feeling relaxed increased physical activity, causing physical discomfort in the following hours. Physical discomfort caused stress as its symptoms resembled her feared somatic anxiety symptoms. This increased the patient's insight that stress, expressed as somatic symptoms, played a central role in her panic disorder. This started a dialogue on how to cope with stress caused by somatic (anxiety) symptoms and provided a rationale for the patient to start an interoceptive exposure intervention she had repeatedly refused before. The observed symptom dynamics may not be generalizable to any other random two weeks. Personalized diagnosis of psychopathology incorporating complex symptom dynamics is feasible and a promising addition to current categorical diagnostic systems and could guide intervention selection. This merits further exploration.


Inhibition of glutamine synthetase in monocytes from patients with acute-on-chronic liver failure resuscitates their antibacterial and inflammatory capacity.
Acute-on-chronic liver failure (ACLF) is associated with dysfunctional circulating monocytes whereby patients become highly susceptible to bacterial infections. Here, we identify the pathways underlying monocyte dysfunction in ACLF and we investigate whether metabolic rewiring reinstates their phagocytic and inflammatory capacity. Following phenotypic characterisation, we performed RNA sequencing on CD14+CD16- monocytes from patients with ACLF and decompensated alcoholic cirrhosis. Additionally, an in vitro model mimicking ACLF patient-derived features was implemented to investigate the efficacy of metabolic regulators on monocyte function. Monocytes from patients with ACLF featured elevated frequencies of interleukin (IL)-10-producing cells, reduced human leucocyte antigen DR isotype (HLA-DR) expression and impaired phagocytic and oxidative burst capacity. Transcriptional profiling of isolated CD14+CD16- monocytes in ACLF revealed upregulation of an array of immunosuppressive parameters and compromised antibacterial and antigen presentation machinery. In contrast, monocytes in decompensated cirrhosis showed intact capacity to respond to inflammatory triggers. Culturing healthy monocytes in ACLF plasma mimicked the immunosuppressive characteristics observed in patients, inducing a blunted phagocytic response and metabolic program associated with a tolerant state. Metabolic rewiring of the cells using a pharmacological inhibitor of glutamine synthetase, partially restored the phagocytic and inflammatory capacity of in vitro generated- as well as ACLF patient-derived monocytes. Highlighting its biological relevance, the glutamine synthetase/glutaminase ratio of ACLF patient-derived monocytes positively correlated with disease severity scores. In ACLF, monocytes feature a distinct transcriptional profile, polarised towards an immunotolerant state and altered metabolism. We demonstrated that metabolic rewiring of ACLF monocytes partially revives their function, opening up new options for therapeutic targeting in these patients.


Exchange of materials between terrestrial ecosystems and the atmosphere.
Many biogenic trace gases are increasing in concentration or flux or both in the atmosphere as a consequence of human activities. Most of these gases have demonstrated or potential effects on atmospheric chemistry, climate, and the functioning of terrestrial ecosystems. Focused studies of the interactions between the atmosphere and the biosphere that regulate trace gases can improve both our understanding of terrestrial ecosystems and our ability to predict regional-and global-scale canges in atmospheric chemistry.


A true branchial fistula in the context of branchiootic syndrome: challenges of diagnosis and management.
The presence of a branchial fistula with communication both internally and externally: a 'true' branchial fistula is rare, and may arise in the context of autosomal dominant conditions such as branchiootic syndrome and branchiootorenal syndrome. We discuss the case of a true branchial fistula, which recurred after initial surgical excision, in a patient with branchiootic syndrome. The residual tract was dissected in a second operation through stepladder neck incisions and removed in toto via an intraoral approach. No renal abnormalities were detected on investigation with ultrasound. Incomplete excision of a branchial sinus is likely to cause recurrence however intraoperative visualisation of the tract can can sometimes prove challenging. An combined intraoral and external approach aids delineation and tract definition when there is a true branchial fistula and can therefore facilitate a complete excision. Suspicion of an hereditary aetiology should be raised in patients with bilateral or preauricular features, or a positive family history, which may then prompt additional renal and genetic investigation.


A colorimetric homogeneous immunoassay system for the C-reactive protein.
The C-reactive protein (CRP), which has a five repeat pentameric structure, is known to be a marker for acute inflammation and a potential risk predictor for cardiovascular disease. A simple and rapid homogeneous assay method for the detection of CRP, based on a gold nanoparticle (AuNP) aggregation induced colorimetric response, has been developed. In the technique, aggregation of CRP antibody-conjugated AuNPs is induced by addition of CRP as a consequence of its unique pentameric structure. CRP-promoted aggregation of the antibody-conjugated AuNPs results in a change of the wavelength maximum in the UV/Vis-spectrum. This homogeneous assay displays a typical hook effect, in which the signal level is directly proportional to CRP concentration until a critical concentration of CRP (the hook point) is reached. Above this concentration, the signal level decreases as the CRP concentration increases. The maximum shift in the absorption maximum was found to occur when the CRP antigen concentration is 100 ng mL(-1). In order to improve the linearity of the method, we employed a procedure, which takes advantage of a saturation phenomenon that leads to the hook effect, to increase the dynamic range of the CRP assay. Specifically, the use of CRP pre-spiked serum promotes maximum aggregation at the low CRP concentrations and, as a result, leads to an increase in the dynamic range for CRP detection. The applicability of the new homogenous assay system was demonstrated by its utilization for qualitative analysis of CRP in serum samples. The combined observations made in this effort show that the method using CRP antibody-conjugated AuNPs is both rapid and simple and, consequently, it can potentially be applied to onsite diagnosis.


Sin3B: an essential regulator of chromatin modifications at E2F target promoters during cell cycle withdrawal.
Efficient and accurate cell cycle exit is intimately linked to cellular differentiation, and by inference, to the prevention of tumorigenesis. Perhaps the most important axis of control for this process involves the interactions of the E2F family of DNA binding proteins with the retinoblastoma (Rb) and Rb-related "pocket protein" (p107 and p130) family of tumor suppressors. Not surprisingly, alterations in this pathway are present in a large number of human malignancies. The molecular basis for the controls exercised by the Rb family of proteins has been widely investigated, but is still not completely understood. Elegant in vitro studies had previously suggested the participation of histone deacetylase (HDAC)-associated Sin3B in E2F-mediated repression. Using genetically modified mice, we have recently uncovered a role for the Sin3B protein as a specific and essential actor in promoting cell cycle exit via the E2F-Rb pathway. We demonstrated its absolute requirement not only for cell cycle exit in vitro and in vivo, but also for biological processes linked to cellular differentiation. These observations strongly suggest that Sin3B plays an essential role in coordinating the chromatin modifying activities required for the transient repression of pro-proliferation genes in quiescence, as well as stable silencing of these genes upon terminal differentiation.


Body mass index and type 2 diabetes and breast cancer survival: a Mendelian randomization study.
The causal relationship between body mass index (BMI) and type 2 diabetes (T2D) and breast cancer prognosis is still ambiguous. The aim of this study was to investigate the prognostic effect of BMI and T2D on breast cancer disease-free survival (DFS) among Asian individuals. In this two-sample Mendelian randomization (MR) study, the instrumental variables (IVs) were identified using a genome-wide association study (GWAS) among 24,000 participants in the Taiwan Biobank. Importantly, the validity of these IVs was confirmed with a previous large-scale GWAS (Biobank Japan Project, BBJ). In this study, we found that a genetic predisposition toward higher BMI (as indicated by BMI IVs, F = 86.88) was associated with poor breast cancer DFS (hazard ratio [HR] = 6.11; P < 0.001). Furthermore, higher level of genetically predicted T2D (as indicated by T2D IVs) was associated with an increased risk of recurrence of and mortality from breast cancer (HR = 1.43; P < 0.001). Sensitivity analyses, including the weighted-median approach, MR-Egger regression, Radial regression and Mendelian randomization pleiotropy residual sum and outlier (MR-PRESSO) supported the consistency of our findings. Finally, the causal relationship between BMI and poor breast cancer prognosis was confirmed in a prospective cohort study. Our MR analyses demonstrated the causal relationship between the genetic prediction of elevated BMI and a greater risk of T2D with poor breast cancer prognosis. BMI and T2D have important clinical implications and may be used as prognostic indicators of breast cancer.


Survey of Red Fluorescence Proteins as Markers for Secretory Granule Exocytosis.
Fluorescent proteins (FPs) have proven to be valuable tools for high-resolution imaging studies of vesicle transport processes, including exo- and endocytosis. Since the pH of the vesicle lumen changes between acidic and neutral during these events, pH-sensitive FPs with near neutral pKa, such as pHluorin, are particularly useful. FPs with pKa>6 are readily available in the green spectrum, while red-emitting pH-sensitive FPs are rare and often not well characterized as reporters of exo- or endocytosis. Here we tested a panel of ten orange/red and two green FPs in fusions with neuropeptide Y (NPY) for use as secreted vesicle marker and reporter of dense core granule exocytosis and release. We report relative brightness, bleaching rate, targeting accuracy, sensitivity to vesicle pH, and their performance in detecting exocytosis in live cells. Tandem dimer (td)-mOrange2 was identified as well-targeted, bright, slowly bleaching and pH-sensitive FP that performed similar to EGFP. Single exocytosis events were readily observed, which allowed measurements of fusion pore lifetime and the dynamics of the exocytosis protein syntaxin at the release site during membrane fusion and cargo release.


The Structural Binding Mode of the Four Autotaxin Inhibitor Types that Differentially Affect Catalytic and Non-Catalytic Functions.
Autotaxin (ATX) is a secreted lysophospholipase D, catalysing the conversion of lysophosphatidylcholine (LPC) to bioactive lysophosphatidic acid (LPA). LPA acts through two families of G protein-coupled receptors (GPCRs) controlling key cellular responses, and it is implicated in many physiological processes and pathologies. ATX, therefore, has been established as an important drug target in the pharmaceutical industry. Structural and biochemical studies of ATX have shown that it has a bimetallic nucleophilic catalytic site, a substrate-binding (orthosteric) hydrophobic pocket that accommodates the lipid alkyl chain, and an allosteric tunnel that can accommodate various steroids and LPA. In this review, first, we revisit what is known about ATX-mediated catalysis, crucially in light of allosteric regulation. Then, we present the known ATX catalysis-independent functions, including binding to cell surface integrins and proteoglycans. Next, we analyse all crystal structures of ATX bound to inhibitors and present them based on the four inhibitor types that are established based on the binding to the orthosteric and/or the allosteric site. Finally, in light of these data we discuss how mechanistic differences might differentially modulate the activity of the ATX-LPA signalling axis, and clinical applications including cancer.


Hormonal modulation of glomerular function.
Glomeruli contain receptors for many hormones. Binding of angiotensin II (ANG II) or antidiuretic hormone (ADH) to glomerular mesangial cells elicits a contractile response. Other hormones induce synthesis of cyclic nucleotides (cAMP, cGMP). Glomeruli also synthesize several prostaglandins, renin, and ANG II. Micropuncture studies in Munich-Wistar rats have examined the effects of vasoactive drugs and hormones on the filtration process. Several vasodilators increase renal plasma flow in the dog and rat, but GFR remains relatively unchanged due to an offsetting fall in the ultrafiltration coefficient (Kf). Vasoconstrictor substances such as ANG II and norepinephrine cause declines in renal plasma flow and Kf, but GFR remains constant due to an increase in the transcapillary hydraulic pressure gradient. Antidiuretic peptides and parathyroid hormone also reduce Kf. Glomerular mesangial cells may regulate Kf by contracting and reducing glomerular capillary surface area. ANG II and ADH directly stimulate mesangial cell contraction in vitro. Other hormones appear to cause contraction by inducing local ANG II synthesis. These hormonal pathways are implicated in the pathogenesis of altered glomerular function in diverse forms of renal injury.


KCNQ2 encephalopathy: A case due to a de novo deletion.
KCNQ2 encephalopathy is characterized by severely abnormal EEG, neonatal-onset epilepsy and developmental delay. It is caused by mutations (typically missense) in the KCNQ2 gene, encoding the voltage gated potassium channel Kv7.2 and leading to a negative-dominant effect. We present one case experiencing recurrent neonatal seizures with changing hemispheres of origin, reminiscent of epilepsy of infancy with migrating focal seizures. At 9months of age the patient is still seizure-free on carbamazepine, although he is developing a spastic-dystonic tetraplegia with severe dysphagia. He harbors a de novo deletion (c.913_915del [p.Phe305del)]), only described once in a couple of severely affected twins, and leading to the deletion of a phenylalanine residue in the pore domain of the channel. In conclusion, our case is the second described with encephalopathy due to this specific deletion (the one and only deletion so far reported in KCNQ2 encephalopathy). Thus, deletion is a newly described mechanism highlighting how not only missense mutations but also deletions in the channel hot spots can lead to a severe phenotype. Furthermore he presented ictal EEG features similar to epilepsy of infancy with migrating focal seizures not previously described.


Classification of genomic sequences via wavelet variance and a self-organizing map with an application to mitochondrial DNA.
We present a new methodology for discriminating genomic symbolic sequences, which combines wavelet analysis and a self-organizing map algorithm. Wavelets are used to extract variation across various scales in the oligonucleotide patterns of a sequence. The variation is quantified by the estimated wavelet variance, which yields a feature vector. Feature vectors obtained from many genomic sequences, possibly of different lengths, are then classified with a nonparametric self-organizing map scheme. When applied to nearly 200 entire mitochondrial DNA sequences, or their fragments, the method predicts species taxonomic group membership very well, and allows the results to be visualized. When only thousands of nucleotides are available, wavelet-based feature vectors of short oligonucleotide patterns are more efficient in discrimination than frequency-based feature vectors of long patterns. This new data analysis strategy could be extended to numeric genomic data. The routines needed to perform the computations are readily available in two packages of software R.


[Successful management of nonocclusive mesenteric ischemia after aortic valve replacement;report of a case].
Non-occlusive mesenteric ischemia (NOMI) is a fatal complication after cardiovascular surgery, but early diagnosis is difficult because the clinical symptoms are not specific. We report a case of NOMI with successful management due to early diagnosis and treatment. A 78-year-old male complained of sudden abdominal pain after aortic valve replacement. NOMI was suspected because his laboratory work-up showed elevated serum transaminase, and computed tomography showed no mesenteric artery obstruction. We started a continuous intravenous infusion of prostaglandin E1, and performed emergency arterial angiography. Since angiography showed vasospasm of the mesenteric artery, we also started a continuous intra-arterial infusion of papaverine. Each vasodilator drug was started within a few hours after the onset of NOMI. His subsequent hospital course was uneventful, and he was discharged without enterectomy or fatal intestinal necrosis.


The role of esophageal pH-impedance testing in clinical practice.
Ambulatory reflux monitoring is typically performed when esophageal symptoms do not respond to usual antireflux medications, or prior to invasive antireflux therapy. Although pH-based metrics have been the standard in defining esophageal reflux burden, novel impedance parameters have been introduced in recent years that can either be extracted from pH-impedance monitoring or obtained directly from esophageal mucosa. This review evaluates the clinical role of esophageal pH-impedance monitoring in clinical practice in the present day. Elevated acid exposure time in the distal esophagus remains the primary metric that predicts symptom improvement from antireflux therapy. Although conventional impedance-based metrics (numbers of reflux episodes, reflux-symptom association) provide complementary evidence, novel impedance-based metrics show promise in documenting reflux-induced damage to esophageal mucosal integrity, and in potentially predicting treatment outcome. The postreflux swallow-induced peristaltic wave measures integrity of primary peristalsis triggered by a reflux episode. Baseline impedance values reflect histopathological damage to mucosal integrity, and improve following successful antireflux therapy. Mucosal impedance assessed at endoscopy is a new diagnostic tool that is currently being evaluated in multinational research studies. Esophageal pH-impedance monitoring has potential to augment confidence in a reflux diagnosis beyond that provided by pH-monitoring alone.


Campylobacteriosis, salmonellosis, and shigellosis in free-ranging human-habituated mountain gorillas of Uganda.
For conservation purposes and due to growing ecotourism, free-ranging mountain gorillas (Gorilla gorilla beringei) have been habituated to humans. Fecal specimens (n = 62) collected in January 1999 from mountain gorillas of the Bwindi and Mgahinga National Parks, Uganda, were tested for Campylobacter spp., Salmonella spp., and Shigella spp., and the overall prevalence of infection was 19%, 13%, and 6%, respectively. The prevalence of positive specimens was not related to the year of habituation of a gorilla group to humans. Campylobacter spp., Salmonella, and Shigella spp. infections were not distributed equally among the age classes of gorillas; most of the enteropathogens (80%), and all Shigella spp. organisms, S. sonnei, S. boydii, and S. flexneri, were isolated from subadults and adult gorillas with ages ranging from 6.0 to 11.9 yr. The prevalence of Campylobacter spp. and Salmonella spp. infections among human-habituated gorillas has doubled during the last 4 yr, and isolation of Shigella spp. for the first time from mountain gorillas, may indicate enhanced anthropozoonotic transmission of these enteropathogens.


Is coeliac disease more prevalent in young adults with coexisting Type 1 diabetes mellitus and autoimmune thyroid disease compared with those with Type 1 diabetes mellitus alone?
It is known that patients with Type 1 diabetes mellitus are more prone to develop coeliac disease and that autoimmune thyroid disease occurs more frequently in patients with coeliac disease. We therefore assessed whether coeliac disease, either known or occult, occurs more frequently in young/middle aged adults with Type 1 diabetes and coexisting autoimmune thyroid dysfunction than in adults with Type 1 diabetes alone. The prevalence of known coeliac disease was assessed in 509 (301 males, aged 16-55 years) patients with Type 1 diabetes, 28 (5.5%) of whom had treated autoimmune thyroid disease. In a second study 38 patients with Type 1 diabetes and coexisting autoimmune thyroid disease along with 112 patients with Type 1 diabetes alone were then screened for coeliac disease using serum IgA endomysial antibodies and IgA gliadin antibodies. Seven of the 509 patients (1.4%) had been diagnosed with coeliac disease and two of these had later developed autoimmune thyroid disease (both hypothyroid). The subsequent screening exercise found that one of the 38 patients with both Type 1 diabetes and thyroid disease had positive endomysial antibodies on screening. However, duodenal biopsy was negative for coeliac disease. There were two patients with positive endomysial antibodies in the group of 112 patients with diabetes only. Both had duodenal biopsy but only one was consistent with coeliac disease. The prevalence of known coeliac disease in this young adult Type 1 diabetes clinic in North-west England was 7/509 (1.4%). Two of these seven patients with coeliac disease were from the group of 28 who had autoimmune thyroid disease as well. Therefore we suggest that patients with known coeliac disease and Type 1 diabetes should be screened for autoimmune thyroid disease. The second screening study then found 3/150 (2%) to have a serological marker for coeliac disease. However, patients with both Type 1 diabetes and autoimmune thyroid disease were not more likely to have occult coeliac disease compared with those with Type 1 diabetes only.


Prognostic significance of minimal residual disease in infants with acute lymphoblastic leukemia treated within the Interfant-99 protocol.
Acute lymphoblastic leukemia (ALL) in infants younger than 1 year is a rare but relatively homogeneous disease ( approximately 80% MLL gene rearranged, approximately 70% CD10-negative) when compared with childhood and adult ALL. Several studies in children and adults with ALL have shown that minimal residual disease (MRD) status is a strong and independent prognostic factor. We therefore evaluated the prognostic significance of MRD in infant ALL. Ninety-nine infant patients treated according to the Interfant-99 protocol were included in this study. MRD was analyzed by real-time quantitative PCR analysis of rearranged immunoglobulin genes, T-cell receptor genes and MLL genes at various time points (TP) during therapy. Higher MRD levels at the end of induction (TP2) and consolidation (TP3) were significantly associated with lower disease-free survival. Combined MRD information at TP2 and TP3 allowed recognition of three patients groups that significantly differed in outcome. All MRD-high-risk patients (MRD levels > or =10(-4) at TP3; 26% of patients) relapsed. MRD-low-risk patients (MRD level <10(-4) at both TP2 and TP3) constituted 44% of patients and showed a relapse-rate of only 13%, whereas remaining patients (MRD-medium-risk patients; 30% of patients) had a relapse rate of 31%. Comparison between the current Interfant-06 stratification at diagnosis and the here presented MRD-based stratification showed that both stratifications recognized different subgroups of patients. These data indicate that MRD diagnostics has added value for recognition of risk groups in infant ALL and that MRD diagnostics can be used for treatment intervention in infant ALL as well.


Sudden-onset tinnitus associated with arterial dissection of the vertebrobasilar system.
Magnetic resonance (MR) images of 62 patients with tinnitus were reviewed. These included T1-weighted and T2-weighted axial images and three-dimensional spoiled gradient-recalled acquisition in steady-state (3D-SPGR) imaging with gadopentetate dimeglumine. In 49 cases with tinnitus occurring gradually, the MR images of 2 cases (4.1%) suggested arterial dissection of the vertebrobasilar system. One of these patients underwent vertebral angiography (VAG) and no dissection was confirmed. In 13 cases of sudden-onset tinnitus, MR images of 3 cases (23.1%) suggested arterial dissection. Of these three cases, VAG was performed in two, resulting in arterial dissections being confirmed in two cases. It was concluded that the arterial dissection of the vertebrobasilar system was one of the causes of sudden-onset tinnitus. Abnormalities in neurotological examinations in cases with tinnitus caused by the arterial dissection of vertebrobasilar system implied that the tinnitus was caused by inner ear dysfunction.


Estimation of Annual Secondary Lung Cancer Deaths Using Various Adjuvant Breast Radiotherapy Techniques for Early-Stage Cancers.
Secondary lung cancer (SLC) can offset the benefit of adjuvant breast radiotherapy (RT), and risks compound sharply after 25 to 30 years. We hypothesized that SLC risk is mainly an issue for early-stage breast cancer, and that lives could be saved using different RT techniques. The SEER database was used to extract breast patient age, stage survival, and radiotherapy utilization over time and per stage and to assess the factors associated with increased SLC risk with a multivariable competing risk Cox model. The number of SLC was calculated using the BEIR model modified with patient survival, age, and use of RT from the SEER database. Stage distribution and number of new breast cancer cases were obtained from the NAACCR. Mean lung dose for various irradiation techniques was obtained from measurement or literature. Out of the 765,697 non-metastatic breast cancers in the SEER database from 1988 to 2012, 49.8% received RT. RT significantly increased the SLC risk for longer follow-up (HR=1.58), early stage including DCIS, stage I and IIA (HR = 1.11), and younger age (HR=1.061) (all p<0.001). More advanced stages did not have significantly increased risk. In 2019, 104,743 early-stage breast patients received radiotherapy, and an estimated 3,413 will develop SLC (3.25%) leading to an excess of 2,900 deaths (2.77%). VMAT would reduce this mortality by 9.9%, hypofractionation 26 Gy in five fractions by 38.8%, a prone technique by 70.3%, 3D-CRT APBI by 43.3%, HDR brachytherapy by 71.1%, LDR by 80.7%, and robotic 4π APBI by 85.2%. SLC after breast RT remains a clinically significant issue for early-stage breast cancers. This mortality could be significantly reduced using a prone technique or APBI.


Hyperexcitable interneurons trigger cortical spreading depression in an Scn1a migraine model.
Cortical spreading depression (CSD), a wave of depolarization followed by depression of cortical activity, is a pathophysiological process implicated in migraine with aura and various other brain pathologies, such as ischemic stroke and traumatic brain injury. To gain insight into the pathophysiology of CSD, we generated a mouse model for a severe monogenic subtype of migraine with aura, familial hemiplegic migraine type 3 (FHM3). FHM3 is caused by mutations in SCN1A, encoding the voltage-gated Na+ channel NaV1.1 predominantly expressed in inhibitory interneurons. Homozygous Scn1aL1649Q knock-in mice died prematurely, whereas heterozygous mice had a normal lifespan. Heterozygous Scn1aL1649Q knock-in mice compared with WT mice displayed a significantly enhanced susceptibility to CSD. We found L1649Q to cause a gain-of-function effect with an impaired Na+-channel inactivation and increased ramp Na+ currents leading to hyperactivity of fast-spiking inhibitory interneurons. Brain slice recordings using K+-sensitive electrodes revealed an increase in extracellular K+ in the early phase of CSD in heterozygous mice, likely representing the mechanistic link between interneuron hyperactivity and CSD initiation. The neuronal phenotype and premature death of homozygous Scn1aL1649Q knock-in mice was partially rescued by GS967, a blocker of persistent Na+ currents. Collectively, our findings identify interneuron hyperactivity as a mechanism to trigger CSD.


Polymyxins for CNS infections: Pharmacology and neurotoxicity.
Central nervous system (CNS) infections caused by multi-drug resistant (MDR) Gram-negative bacteria present a major health and economic burden worldwide. Due to the nearly empty antibiotic discovery pipeline, polymyxins (i.e. polymyxin B and colistin) are used as the last-line therapy against Gram-negative 'superbugs' when all other treatment modalities have failed. The treatment of CNS infections due to multi-drug resistant Gram-negative bacteria is problematic and associated with high mortality rates. Colistin shows significant efficacy for the treatment of CNS infections caused by MDR Gram-negative bacteria that are resistant to all other antibiotics. In particular, MDR Acinetobacter baumannii, Pseudomonas aeruginosa and Klebsiella pneumoniae which are resistant to expanded-spectrum and fourth-generation cephalosporins, carbapenems and aminoglycosides, represent a major therapeutic challenge, although they can be treated with colistin or polymyxin B. However, current dosing recommendations of intrathecal/intraventricular polymyxins are largely empirical, as we have little understanding of the pharmacokinetics/pharmacodynamics and, importantly, we are only starting to understand the mechanisms of potential neurotoxicity. This review covers the current knowledge-base on the mechanisms of disposition and potential neurotoxicity of polymyxins as well as the combined use of neuroprotective agents to alleviate polymyxins-related neurotoxicity. Progress in this field will provide the urgently needed pharmacological information for safer and more efficacious intrathecal/intraventricular polymyxin therapy against life-threatening CNS infections caused by Gram-negative 'superbugs'.


IMAGe syndrome: clinical and genetic implications based on investigations in three Japanese patients.
Arboleda et al. have recently shown that IMAGe (intra-uterine growth restriction, metaphyseal dysplasia, adrenal hypoplasia congenita and genital abnormalities) syndrome is caused by gain-of-function mutations of maternally expressed gene CDKN1C on chromosome 11p15.5. However, there is no other report describing clinical findings in patients with molecularly studied IMAGe syndrome. Here, we report clinical and molecular findings in Japanese patients. We studied a 46,XX patient aged 8·5 years (case 1) and two 46,XY patients aged 16·5 and 15·0 years (cases 2 and 3). Clinical studies revealed not only IMAGe syndrome-compatible phenotypes in cases 1-3, but also hitherto undescribed findings including relative macrocephaly and apparently normal pituitary-gonadal endocrine function in cases 1-3, familial glucocorticoid deficiency (FGD)-like adrenal phenotype and the history of oligohydramnios in case 2, and arachnodactyly in case 3. Sequence analysis of CDKN1C, pyrosequencing-based methylation analysis of KvDMR1 and high-density oligonucleotide array comparative genome hybridization analysis for chromosome 11p15.5 were performed, showing an identical de novo and maternally inherited CDKN1C gain-of-function mutation (p.Asp274Asn) in cases 1 and 2, respectively, and no demonstrable abnormality in case 3. The results of cases 1 and 2 with CDKN1C mutation would argue the following: [1] relative macrocephaly is consistent with maternal expression of CDKN1C in most tissues and biparental expression of CDKN1C in the foetal brain; [2] FGD-like phenotype can result from CDKN1C mutation; and [3] genital abnormalities may primarily be ascribed to placental dysfunction. Furthermore, lack of CDKN1C mutation in case 3 implies genetic heterogeneity in IMAGe syndrome.


Functional Brain Changes During Mindfulness-Based Cognitive Therapy Associated With Tinnitus Severity.
Mindfulness-based therapies have been introduced as a treatment option to reduce the psychological severity of tinnitus, a currently incurable chronic condition. This pilot study of twelve subjects with chronic tinnitus investigates the relationship between measures of both task-based and resting state functional magnetic resonance imaging (fMRI) and measures of tinnitus severity, assessed with the Tinnitus Functional Index (TFI). MRI was measured at three time points: before, after, and at follow-up of an 8-week long mindfulness-based cognitive therapy intervention. During the task-based fMRI with affective sounds, no significant changes were observed between sessions, nor was the activation to emotionally salient compared to neutral stimuli significantly predictive of TFI. Significant results were found using resting state fMRI. There were significant decreases in functional connectivity among the default mode network, cingulo-opercular network, and amygdala across the intervention, but no differences were seen in connectivity with seeds in the dorsal attention network (DAN) or fronto-parietal network and the rest of the brain. Further, only resting state connectivity between the brain and the amygdala, DAN, and fronto-parietal network significantly predicted TFI. These results point to a mostly differentiated landscape of functional brain measures related to tinnitus severity on one hand and mindfulness-based therapy on the other. However, overlapping results of decreased amygdala connectivity with parietal areas and the negative correlation between amygdala-parietal connectivity and TFI is suggestive of a brain imaging marker of successful treatment.


Non-surgical Management of Congenital Auricular Deformities.
Unlike congenital auricular malformations which are identified by underdevelopment of dermal and cartilaginous tissues, deformed ears are less sever congenital anomalies characterized only by a misshaped pinna structure and can be improved with acceptable cosmetic results and minimal cost through ear molding if treated in early neonatal period. In this study, authors present the first report of using splinting techniques for treatment of deformational auricular anomalies in Iranian children. Our case load consisted of a series of 29 patients (Male=16, Female=13) who were referred to Plastic Surgery Unit of Shiraz University of Medical Sciences from September 2011 to December 2014. Children aged more than 6 moths were excluded. Twenty-nine children affected by various deformities including prominent ears (n=11), lop ears (n=8) and constricted ears (n=10) were treated by splintage as a nonsurgical technique. The mean time of treatment was 13.33±2 weeks. Eight (27.6%) patients did not complete the treatment. Splinting resulted in excellent or satisfactory results in 12 (57.14%) of treated cases. No improvement was observed at the end of the molding treatment in 9 patients. No complication was observed during the treatment in any of the patients. The nonsurgical molding can be used as an effective approach for achieving natural outcomes and correcting cosmetic abnormalities. Rate of satisfaction is dependent on type of deformity, the neonatal age in which treatment started and also parents' adherence to treatment methods and principals. Concerning the low rate of complications and high satisfactory results the method can be used instead of surgery in appropriate cases.


The deleterious effects of arteriovenous fistula-creation on the cardiovascular system: a longitudinal magnetic resonance imaging study.
Arteriovenous fistula-formation remains critical for the provision of hemodialysis in end-stage renal failure patients. Its creation results in a significant increase in cardiac output, with resultant alterations in cardiac stroke volume, systemic blood flow, and vascular resistance. The impact of fistula-formation on cardiac and vascular structure and function has not yet been evaluated via "gold standard" imaging techniques in the modern era of end-stage renal failure care. A total of 24 patients with stage 5 chronic kidney disease undergoing fistula-creation were studied in a single-arm pilot study. Cardiovascular magnetic resonance imaging was undertaken at baseline, and prior to and 6 months following fistula-creation. This gold standard imaging modality was used to evaluate, via standard brachial flow-mediated techniques, cardiac structure and function, aortic distensibility, and endothelial function. At follow up, left ventricular ejection fraction remained unchanged, while mean cardiac output increased by 25.0% (P<0.0001). Significant increases in left and right ventricular end-systolic volumes (21% [P=0.014] and 18% [P<0.01]), left and right atrial area (11% [P<0.01] and 9% [P<0.01]), and left ventricular mass were observed (12.7% increase) (P<0.01). Endothelial-dependent vasodilation was significantly decreased at follow up (9.0%±9% vs 3.0%±6%) (P=0.01). No significant change in aortic distensibility was identified. In patients with end-stage renal failure, fistula-formation is associated with an increase in cardiac output, dilation of all cardiac chambers and deterioration in endothelial function.


Mutations in the Neuroblastoma Amplified Sequence gene in a family affected by Acrofrontofacionasal Dysostosis type 1.
Acrofrontofacionasal Dysostosis type 1 (AFFND1) is an extremely rare, autosomal recessive syndrome, comprising facial and skeletal abnormalities, short stature and intellectual disability. We analyzed an Indian family with two affected siblings by exome sequencing and identified a novel homozygous truncating mutation in the Neuroblastoma-Amplified Sequence (NBAS) gene in the patients' genome. Mutations in the NBAS gene have recently been associated with different phenotypes mainly involving skeletal formation, liver and cognitive development. The NBAS protein has been implicated in two key cellular processes, namely the non-sense mediated decay and the Golgi-to-Endoplasmic Reticulum retrograde traffic. Both functions were impaired in HEK293T cells overexpressing the truncated NBAS protein, as assessed by Real-Time PCR, Western blot analysis, co-immunoprecipitation, and immunofluorescence analysis. We examined the expression of NBAS protein in mouse embryos at various developmental stages by immunohistochemistry, and detected expression in developing chondrogenic and osteogenic structures of the skeleton as well as in the cortex, hippocampus and cerebellum, which is compatible with a role in bone and brain development. Functional genetics in the zebrafish model showed that depletion of endogenous z-nbas in fish embryos results in defective morphogenesis of chondrogenic cranial skeletal elements. Overall, our data point to a conserved function of NBAS in skeletal morphogenesis during development, support the hypothesis of a causative role of the mutated NBAS gene in the pathogenesis of AFFND1 and extend the spectrum of phenotypes associated with defects in this gene.


N-Palmitoylethanolamide Exerts Antidepressant-Like Effects in Rats: Involvement of PPARα Pathway in the Hippocampus.
N-Palmitoylethanolamide (PEA), an endocannabinoid-like molecule, participates in controlling behaviors associated with mental disorders as an endogenous neuroprotective factor. On the basis of accumulating evidence and our previous data, we tested the hypothesis that the antidepressant-like effects of PEA observed during chronic unpredictable mild stress (CUMS) are mediated by possible targets in the peroxisome proliferator-activated receptor alpha (PPARα) pathway. In this study, rats were subjected to 35 days of CUMS and treated with drugs such as PEA (2.5, 5.0, or 10 mg/kg, by mouth), fluoxetine (10 mg/kg, by mouth), or the combination of PEA and MK886 (1-[(4-chlorophenyl) methyl]-3-[(1,1-dimethylethyl) thio]-α,α-dimethyl-5-(1-methylethyl)-1H-indole-2-propanoic acid). After behavioral tests, the animals were sacrificed and their hippocampi were dissected for subsequent studies. PEA normalized weight gain, sucrose preferences, locomotor activity in an open-field test, and levels of the PPARα mRNA and protein in the hippocampus, and it reduced serum adrenocorticotropic hormone (ACTH) and corticosterone (CORT) levels in rats subjected to CUMS. PEA reversed the abnormal levels of several oxidative stress biomarkers and increased the concentrations of two neurotrophic factors in the hippocampus of CUMS-induced rats. In addition, PEA alleviated the decrease in hippocampal weight. However, the aforementioned effects of PEA were completely or partially abolished by MK886, a selective PPARα antagonist. On the basis of these findings, the PPARα pathway in the hippocampus is a possible target of the antidepressant effects of PEA, and the maintenance of a stable hypothalamic-pituitary-adrenal axis, the antioxidant defenses, and normalization of neurotrophic factor levels in the hippocampus are involved in this process.


Loss of succinyl-CoA synthase ADP-forming β subunit disrupts mtDNA stability and mitochondrial dynamics in neurons.
Succinyl Coenzyme A synthetase (SCS) is a key mitochondrial enzyme. Defected SCS ADP-forming β subunit (SCS A-β) is linked to lethal infantile Leigh or leigh-like syndrome. However, the impacts of SCS A-β deficiency on mitochondria specifically in neurons have not yet been comprehensively investigated. Here, by down-regulating the expression levels of SCS A-β in cultured mouse neurons, we have found that SCS A-β deficiency induces severe mitochondrial dysfunction including lowered oxidative phosphorylation (OXPHOS) efficiency, increased mitochondrial superoxide production, and mtDNA depletion as well as aberrations of mitochondrial fusion and fission proteins, which eventually leads to neuronal stress. Our data also suggest that the deregulation of mitochondrial nucleoside diphosphate kinase (NDPK) together with defects in mitochondrial transcription factors including mitochondrial DNA pol γ and Twinkle contribute to SCS A-β deficiency-mediated mtDNA instability. Furthermore, we have found that SCS A-β deficiency has detrimental influence on neuronal mitochondrial dynamics. Put together, the results have furnished our knowledge on the pathogenesis of SCS A-β deficiency-related mitochondrial diseases and revealed the vital role of SCS A-β in maintaining neuronal mitochondrial quality control and neuronal physiology.


PPAR alpha, more than PPAR delta, mediates the hepatic and skeletal muscle alterations induced by the PPAR agonist GW0742.
Therapeutic use of certain peroxisome proliferator-activated receptor (PPAR) alpha agonists (fibrates) for the treatment of dyslipidemia has infrequently been associated with the untoward side effect of myopathy. With interest in PPAR-delta as a therapeutic target, this study assessed whether a PPAR-delta agonist induced similar hepatic and skeletal muscle alterations as noted with some fibrates. PPAR-alpha null (KO) and corresponding wild-type (WT) mice were administered toxicological dosages of a potent PPAR-delta agonist tool ligand (GW0742; which also has weak PPAR-alpha agonist activity) or a potent PPAR-alpha agonist (WY-14,643) for 10 days. Increases in liver weights and clinical chemistry indicators of skeletal muscle damage and/or liver injury were more pronounced in WT mice compared with KO mice administered the PPAR-delta agonist. Likewise, the incidence and severity of skeletal myopathy were greater in WT mice given GW0742 compared with KO mice. Ultrastructural and immunohistochemical analyses revealed significant peroxisome proliferation in muscle and liver of WT mice treated with each agonist; however, KO animals showed little or no evidence of hepatic and muscle peroxisome proliferation. PMP-70 protein expression in liver was consistent with these results. The hepatomegaly, hepatic and skeletal muscle peroxisome proliferation, and skeletal myopathy induced by this PPAR-delta ligand was predominantly mediated by its cross-activation of PPAR-alpha, though PPAR-delta agonism contributed slightly to these effects.


Rapid elicitation of broadly neutralizing antibodies to HIV by immunization in cows.
No immunogen to date has reliably elicited broadly neutralizing antibodies to HIV in humans or animal models. Advances in the design of immunogens that antigenically mimic the HIV envelope glycoprotein (Env), such as the soluble cleaved trimer BG505 SOSIP, have improved the elicitation of potent isolate-specific antibody responses in rabbits and macaques, but so far failed to induce broadly neutralizing antibodies. One possible reason for this failure is that the relevant antibody repertoires are poorly suited to target the conserved epitope regions on Env, which are somewhat occluded relative to the exposed variable epitopes. Here, to test this hypothesis, we immunized four cows with BG505 SOSIP. The antibody repertoire of cows contains long third heavy chain complementary determining regions (HCDR3) with an ultralong subset that can reach more than 70 amino acids in length. Remarkably, BG505 SOSIP immunization resulted in rapid elicitation of broad and potent serum antibody responses in all four cows. Longitudinal serum analysis for one cow showed the development of neutralization breadth (20%, n = 117 cross-clade isolates) in 42 days and 96% breadth (n = 117) at 381 days. A monoclonal antibody isolated from this cow harboured an ultralong HCDR3 of 60 amino acids and neutralized 72% of cross-clade isolates (n = 117) with a potent median IC50 of 0.028 μg ml-1. Breadth was elicited with a single trimer immunogen and did not require additional envelope diversity. Immunization of cows may provide an avenue to rapidly generate antibody prophylactics and therapeutics to address disease agents that have evolved to avoid human antibody responses.


Influence of long-range interactions on quantum interference in molecular conduction. A tight-binding (Hückel) approach.
When certain pairs of atoms in a π-conjugated molecule are connected with nanometer-scale source and drain electrodes, the remarkable quantum interference (QI) effect may arise. In this case, the electron transmission probability is significantly suppressed due to the QI effect. Tight-binding approaches, such as the Hückel molecular orbital (HMO) model, have revealed important features of this quantum phenomenon. However, important deviations from experiments and from more sophisticated calculations are known for a variety of cases. Here, we propose an extension of the HMO method to include non-nearest-neighbor interactions. Such long-range interactions (LRIs) are implemented in the HMO model in the form of a damping function that decays as the topological distance-the number of bonds separating two atoms-gets larger. The proposed model is further developed so that a geometric modification, i.e., the rotation around a single bond, can be taken into account. Our results show that LRI affects both the location of the antiresonance peak due to QI and the intensity of QI, even suppressing it in some cases. These results agree well with what was observed in a Density Functional based Tight-Binding (DFTB) study reported in the literature. These properties can be interpreted on the basis of a graph-theoretic path-counting model as well as the molecular orbital theory. In addition, the geometric LRI model is shown to reproduce the change of transmission as a function of rotation around the single bond separating two benzene rings in biphenyl, in agreement with what was observed in both experiment and DFTB calculation.


Immobilizing nitrogen to control plant invasion.
Increased soil N availability may often facilitate plant invasions. Therefore, lowering N availability might reduce these invasions and favor desired species. Here, we review the potential efficacy of several commonly proposed management approaches for lowering N availability to control invasion, including soil C addition, burning, grazing, topsoil removal, and biomass removal, as well as a less frequently proposed management approach for lowering N availability, establishment of plant species adapted to low N availability. We conclude that many of these approaches may be promising for lowering N availability by stimulating N immobilization, even though most are generally ineffective for removing N from ecosystems (excepting topsoil removal). C addition and topsoil removal are the most reliable approaches for lowering N availability, and often favor desired species over invasive species, but are too expensive or destructive, respectively, for most management applications. Less intensive approaches, such as establishing low-N plant species, burning, grazing and biomass removal, are less expensive than C addition and may lower N availability if they favor plant species that are adapted to low N availability, produce high C:N tissue, and thus stimulate N immobilization. Regardless of the method used, lowering N availability sufficiently to reduce invasion will be difficult, particularly in sites with high atmospheric N deposition or agricultural runoff. Therefore, where feasible, the disturbances that result in high N availability should be limited in order to reduce invasions by nitrophilic weeds.


The role of taste in alcohol preference, consumption and risk behavior.
Alcohol consumption is widespread, and high levels of use are associated with increased risk of developing an alcohol use disorder. Thus, understanding the factors that influence alcohol intake is important for disease prevention and management. Additionally, elucidating the factors that associate with alcohol preference and intake in non-clinical populations allows for product development and optimisation opportunities for the alcoholic beverage industry. The literature on how taste (orosensation) influences alcohol behavior is critically appraised in this review. Ethanol, the compound common to all alcoholic beverages, is generally aversive as it primarily elicits bitterness and irritation when ingested. Individuals who experience orosensations (both taste and chemesthetic) more intensely tend to report lower liking and consumption of alcoholic beverages. Additionally, a preference for sweetness is likely associated with a paternal history of alcohol use disorders. However, conflicting findings in the literature are common and may be partially attributable to differences in the methods used to access orosensory responsiveness and taste phenotypes. We conclude that while taste is a key driver in alcohol preference, intake and use disorder, no single taste-related factor can adequately predict alcohol behaviour. Areas for further research and suggestions for improved methodological and analytical approaches are highlighted.


Bilateral styloid fracture. A case report.
Styloid fractures are common, and their treatment is relatively straightforward. This case demonstrated delayed healing because of continuous pathologic motion aggravating the styloid fractures.


Effect of indole-3-butyric acid on phytoplasmas in infected Catharanthus roseus shoots grown in vitro.
Phytoplasmas are noncultivable bacteria usually maintained in Catharanthus roseus shoots grown in vitro on MS medium with benzylaminopurine. The aim of our research was to examine the influence of indole-3-butyric acid (IBA) on C. roseus shoots infected with three different phytoplasma strains. Supplement of IBA in the medium supported plant growth, photosynthesis and remission of symptoms in all phytoplasma-infected shoots, but had no effect on the presence of EY-C and SA-I phytoplasma strains in tested tissue. However, HYDB phytoplasma was undetectable in approximately half of the tested shoots grown on the medium with IBA. After 1 year of IBA treatment, HYDB-infected periwinkle shoots were retransferred to the medium supplemented with benzylaminopurine. Some of the shoots showing remission of symptoms during the IBA treatment permanently escaped the infection and remained negative when tested for phytoplasma presence. This is the first report on the differential influence of plant growth regulators on phytoplasma-infected C. roseus shoots.


Seeing the sound of castanets: Acoustic resonances between shells captured by high-speed optical visualization with 1-mm resolution.
Sound fields radiated from the castanet, a Spanish percussive instrument comprising two shells, were optically visualized. A measurement system, which used parallel phase-shifting interferometry and a high-speed polarization camera, enabled the capture of instantaneous sound fields around the castanets, while the castanets were played, with the spatial resolution of 1.1 mm and frame rate of 100 000 fps. By carefully aligning the tilt of the castanets, the sound fields within the 1-mm gaps between both the shells were captured. From the visualization results, two acoustic resonances between the shells were identified. The first mode appeared between 1000 and 2000 Hz and exhibited a frequency chirp of several hundred hertz for several milliseconds after the impact. This can be explained by the Helmholtz resonance with a time-varying resonator shape, which is caused by the movement of the shells after impact. The second mode showed a resonance pattern with a single nodal diameter at the center of the shells, i.e., the standing wave mode caused by the interior volume. These physical phenomena involved in the sound radiation were identified owing to the unique features of the optical imaging method, such as contactless nature and millimeter-resolution imaging of instantaneous pressure fields.


Influence of lung parenchymal destruction on the different indexes of the methacholine dose-response curve in COPD patients.
The interpretation of nonspecific bronchial provocation dose-response curves in COPD is still a matter of debate. Bronchial hyperresponsiveness (BHR) in patients with COPD could be influenced by the destruction of the parenchyma and the augmented mechanical behavior of the lung. Therefore, we studied the interrelationships between indexes of BHR, on the one hand, and markers of lung parenchymal destruction, on the other. COPD patients were selected by clinical symptoms, evidence of chronic, nonreversible airways obstruction, and BHR, which was defined as a provocative dose of a substance (histamine) causing a 20% fall in FEV(1) (PC(20)) of </= 8 mg/mL. BHR was subsequently studied by methacholine dose-response curves to which a sigmoid model was fitted for the estimation of plateau values and reactivity. Model fits of quasi-static lung pressure-volume (PV) curves yielded static lung compliance (Cstat), the exponential factor (KE) and elastic recoil at 90% of total lung capacity (P90TLC). Carbon monoxide (CO) transfer was measured with the standard single-breath method. Twenty-four patients were included in the study, and reliable PV data could be obtained from 19. The following mean values ( +/- SD) were taken: FEV(1), 65 +/- 12% of predicted; reversibility, 5.6 +/- 3.1% of predicted; the PC(20) for methacholine, 4.3 +/- 5.2 mg/mL; reactivity, 11.0 +/- 5.6% FEV(1)/doubling dose; plateau, 48.8 +/- 17.4% FEV(1); transfer factor, 76.7 +/- 17.9% of predicted; transfer coefficient for carbon monoxide (KCO), 85.9 +/- 22.6% of predicted; Cstat, 4.28 +/- 2.8 kPa; shape factor (KE), 1.9 +/- 1.5 kPa; and P90TLC, 1.1 +/- 0.8 kPa. We confirmed earlier reported relationships between Cstat, on the one hand, and KE (p < 0.0001), P90TLC (p = 0.0012), and KCO percent predicted (p = 0.006), on the other hand. The indexes of the methacholine provocation test were not related to any parameter of lung elasticity and CO transfer. BHR in COPD patients who smoke most probably is determined by airways pathology rather than by the augmented mechanical behavior caused by lung parenchymal destruction.


Effect of temperature, elevated carbon dioxide, and drought during seed development on the isoflavone content of dwarf soybean [Glycine max (L.) Merrill] grown in controlled environments.
The effects of elevated temperature, carbon dioxide, and water stress on the isoflavone content of seed from a dwarf soybean line [Glycine max (L.) Merrill] were determined, using controlled environment chambers. Increasing the temperature from 18 degrees C during seed development to 23 degrees C decreased total isoflavone content by about 65%. A further 5 degrees C increase to 28 degrees C decreased the total isoflavone content by about 90%. Combining treatments at elevated temperature with elevated CO(2) (700 ppm) and water stress to determine the possible consequences of global climate change on soybean seed isoflavone content indicated that elevated CO(2) at elevated temperatures could partially reverse the effects of temperature on soybean seed isoflavone content. The addition of drought stress to plants grown at 23 degrees C and elevated CO(2) returned the total isoflavone levels to the control values obtained at 18 degrees C and 400 ppm CO(2). The promotive effects of drought and elevated CO(2) at 23 degrees C on the 6' '-O-malonygenistin and genistin levels were additive. The individual isoflavones often had different responses to the various growth conditions during seed maturation, modifying the proportions of the principal isoflavones. Therefore, subtle changes in certain environmental factors may change the isoflavone content of commercially grown soybean, altering the nutritional values of soy products.


Monostotic fibrous dysplasia.
Fibrous dysplasia is most often monostotic and an incidental finding in an asymptomatic patient. Familiarity with the disease can promote early diagnosis and avoid lengthy evaluation. The polyostotic form of the disease and its variants, however, can have significant impact on the patient and is associated with several well-defined syndromes. Imaging is central for diagnosis and treatment planning. A wide spectrum of imaging findings includes the characteristic "ground-glass" pattern of mineralization and medullary expansion. Surgical intervention may be necessary to manage secondary complications and deformities.


Impact of liver tumour burden, alkaline phosphatase elevation, and target lesion size on treatment outcomes with 177Lu-Dotatate: an analysis of the NETTER-1 study.
To assess the impact of baseline liver tumour burden, alkaline phosphatase (ALP) elevation, and target lesion size on treatment outcomes with 177Lu-Dotatate. In the phase 3 NETTER-1 trial, patients with advanced, progressive midgut neuroendocrine tumours (NET) were randomised to 177Lu-Dotatate (every 8 weeks, four cycles) plus octreotide long-acting release (LAR) or to octreotide LAR 60 mg. Primary endpoint was progression-free survival (PFS). Analyses of PFS by baseline factors, including liver tumour burden, ALP elevation, and target lesion size, were performed using Kaplan-Meier estimates; hazard ratios (HRs) with corresponding 95% CIs were estimated using Cox regression. Significantly prolonged median PFS occurred with 177Lu-Dotatate versus octreotide LAR 60 mg in patients with low (< 25%), moderate (25-50%), and high (> 50%) liver tumour burden (HR 0.187, 0.216, 0.145), and normal or elevated ALP (HR 0.153, 0.177), and in the presence or absence of a large target lesion (diameter > 30 mm; HR, 0.213, 0.063). Within the 177Lu-Dotatate arm, no significant difference in PFS was observed amongst patients with low/moderate/high liver tumour burden (P = 0.7225) or with normal/elevated baseline ALP (P = 0.3532), but absence of a large target lesion was associated with improved PFS (P = 0.0222). Grade 3 and 4 liver function abnormalities were rare and did not appear to be associated with high baseline liver tumour burden. 177Lu-Dotatate demonstrated significant prolongation in PFS versus high-dose octreotide LAR in patients with advanced, progressive midgut NET, regardless of baseline liver tumour burden, elevated ALP, or the presence of a large target lesion. Clinicaltrials.gov : NCT01578239, EudraCT: 2011-005049-11.


Uric acid and angiotensin II additively promote inflammation and oxidative stress in human proximal tubule cells by activation of toll-like receptor 4.
Renal proximal tubular cells (PTECs) participate in several mechanisms of innate immunity, express toll-like receptors (TLRs), and proinflammatory cytokines. Hyperuricemia may be a promoter of inflammation and renal damage. Angiotensin II (Ang II) modulate immune and inflammatory responses in renal tubular cells. With the aim to evaluate the effect of uric acid (UA) and Ang II on oxidative stress and inflammation mediated by toll-like receptor 4 (TLR4) activation in human PTECs, human kidney 2 (HK2) were incubated for 24 hr with UA (12 mg/dl) and Ang II (10 -7 M). HK2 were pretreated with an antagonist of TLR4 (TAK 242), valsartan or losartan. The genic expression of TLR4, monocyte chemoattractant protein-1 (MCP1), and Nox4 was quantified with reverse transcription polymerase chain reaction, proteins were evaluated with Western blot. The incubation of HK2 either with UA or with Ang II determines an increased expression of TLR4, production of proinflammatory cytokines as MCP1 and pro-oxidants as Nox4 ( p < 0.05). TAK 242 attenuates the expression of MCP1 induced both by UA and Ang II. Valsartan attenuated all the effects we described after exposure to Ang II but not those observed after UA exposure. At variance, pretreatment with losartan, which inhibits UA internalization, attenuates the expression of TLR4, MCP1, and Nox4 in cells previously treated with UA, Ang II, and UA plus Ang II. Proinflammatory pathways are induced in an additive manner by UA and Ang II ( p < 0.05) and might be mediated by TLR4 in PTECs. Renin-angiotensin-aldosterone system (RAAS) activation, hyperuricemia, and innate immunity interplay in the development of chronic tubular damage and the interaction of several nephrotoxic mechanisms blunt the protective effect of RAAS inhibition.


Novel acoustic technology for studying free-ranging shark social behaviour by recording individuals' interactions.
Group behaviours are widespread among fish but comparatively little is known about the interactions between free-ranging individuals and how these might change across different spatio-temporal scales. This is largely due to the difficulty of observing wild fish groups directly underwater over long enough time periods to quantify group structure and individual associations. Here we describe the use of a novel technology, an animal-borne acoustic proximity receiver that records close-spatial associations between free-ranging fish by detection of acoustic signals emitted from transmitters on other individuals. Validation trials, held within enclosures in the natural environment, on juvenile lemon sharks Negaprion brevirostris fitted with external receivers and transmitters, showed receivers logged interactions between individuals regularly when sharks were within 4 m ( approximately 4 body lengths) of each other, but rarely when at 10 m distance. A field trial lasting 17 days with 5 juvenile lemon sharks implanted with proximity receivers showed one receiver successfully recorded association data, demonstrating this shark associated with 9 other juvenile lemon sharks on 128 occasions. This study describes the use of acoustic underwater proximity receivers to quantify interactions among wild sharks, setting the scene for new advances in understanding the social behaviours of marine animals.


New approaches to trials in glomerulonephritis.
Randomized controlled trials are required to reliably identify interventions to improve the outcomes for people with glomerulonephritis (GN). Unfortunately, although easier, observational studies are inherently unreliable even though the findings of both study designs agree most of the time. Currently there are ∼790 trials in GN, but suboptimal design and reporting, together with small sample sizes, mean that they may not be reliable for decision making. If the history is somewhat bleak, the future looks bright, with recent initiatives to improve the quality, size and relevance of clinical trials in nephrology, including greater patient engagement, trial networks, core outcome sets, registry-based trials and adaptive designs. Given the current state of the evidence informing the care of people with GN, disruptive technologies and pervasive culture change is required to ensure that the potential of trials to improve the health of people with this complex condition is to be realized.


Importance of truncal-type occlusion in stentriever-based thrombectomy for acute stroke.
To investigate whether angiographically defined occlusion type could predict of the etiology of acute intracranial large artery occlusion and the stentriever response. We reviewed consecutive patients with acute intracranial large artery occlusion who underwent endovascular treatment and examined their workups for embolic sources. Patient demographics, laboratory findings, hyperdense artery sign, and angiographic occlusion type (truncal-type or branching-site occlusion) were compared between embolic sources (+) and (-) groups. These variables were also compared between stentriever failure and success groups. Details of endovascular procedures were also compared according to occlusion type. A total of 259 patients (mean age 70.3 years; M:F = 132:127) were finally included. Of these patients, 216 (83.4%) were assigned to the embolic sources (+) group after thorough evaluation. Young age, no coronary artery disease, and truncal-type occlusion (odds ratio [OR] 9.07; 95% confidence interval [CI] 3.74-22.0) were independently associated with the embolic source (-) group. Of the overall group, 224 patients (86.5%) underwent stentriever-based endovascular treatment. Hypertension, diabetes, high C-reactive protein level, and truncal-type occlusion (OR 32.2; 95% CI 7.78-133.0) were independent predictors of stentriever failure. Truncal-type occlusion was associated with more reocclusion (77.3% vs 5.0%), resulting in recanalization failure by the stentriever (81.8% vs 20.3%), a longer puncture-to-recanalization time (118.0 vs 49.5 minutes), and more rescue treatment for final successful recanalization (78.9% vs 7.0%). Angiographic occlusion type is an independent predictor of stentriever refractoriness and of the underlying stroke mechanism.


[Species and quantitative characteristics of pharyngeal mucosa microflora in pregnant women].
Species and quantitative characteristics of upper respiratory tract (URT) mucosa microflora in women at gestation period. The results of a bacteriological study of 68 samples of mucus from posterior pharyngeal wall in women at gestation period (from 14 weeks), 52 of those--from pregnant women with acute respiratory infection (ARI) symptoms and 16--from women without signs of disease, are presented. Qualitative and quantitative composition of microflora was studied by a generally accepted bacteriological method. During primary bacteriological study 111 microorganism cultures were isolated. 88 (79.3%) of strains belonged to Gram-positive flora, 20 (18.0%)--to Gram-negative, and Candida genus fungi constituted 3 (2.7%) isolates. Streptococcus pyogenes and Moraxella catarrhalis were isolated from pregnant women with ARI signs at 23.1% and 5.8% frequency of occurrence, respectively. A higher detectability of Staphylococcus aureus--in 31.3% and Candida spp.--in 6.3% of women who did not complain as opposed to patients with URT lesions (in 21.2 and 3.9%, respectively) was determined. In patients without ARI signs the amount of bacteria did not exceed 10(5)--10(6) CFU/ml, in pregnant women with ARI diagnosis in 8 of 52 cases semination of pharyngeal mucuswas observed--10(7)--10(8) CFU/ml. Prevalence of S. aureus, Streptococcus agalactiae, S. pyogenes, Streptococcus mutans in composition of pharyngeal mucus microflora of pregnant women both with URT lesion signs and without them was shown, however the degree of semination by pathogens in the groups was different that determined the severity of disease manifestations.


Blood stains of the Turin Shroud 2015: beyond personal hopes and limitations of techniques.
In the early '80s, evidence was provided that, rather than a dye (red okra), hemoglobin was indeed responsible for the alleged blood stains of the Turin Shroud. Such stains were shown to belong to an MNS positive individual of the AB group, and the halos surrounding the blood stains were compatible with serum containing trace amounts of bilirubin, albumin and immunoglobulins. However, being only based on indirect and circumstantial evidence, most of these data were challenged. In the late '90s, together with the evidence of the gene coding β-globin, contamination between male and female DNA was documented on the Turin Shroud. Although the presence of male was more noticeable than female DNA, these data were considered null and void. These days, to establish that blood indisputably belongs to an MNS positive individual of the AB group, and to exclude DNA contamination, high-specificity techniques with monoclonal antibodies and molecular studies on nuclear and mitochondrial DNA are needed. Indeed, consistent with DNA contamination on the Turin Shroud, sequences from multiple subjects of different ethnic origins have been recently detected on the human mitochondrial genome extracted from dust particles of the linen. Innovative concepts are likely to come up using modern research approaches to evaluate the issue of blood stains of the Turin Shroud. Nor can we rule out the possibility that religious implications of the new findings on the Turin Shroud might be envisaged. Conceivably enough, the ongoing debate will be fierce and passionate, especially in the media.


3-Hydroxyanthralinic acid metabolism controls the hepatic SREBP/lipoprotein axis, inhibits inflammasome activation in macrophages, and decreases atherosclerosis in Ldlr-/- mice.
Atherosclerosis is a chronic inflammatory disease involving immunological and metabolic processes. Metabolism of tryptophan (Trp) via the kynurenine pathway has shown immunomodulatory properties and the ability to modulate atherosclerosis. We identified 3-hydroxyanthranilic acid (3-HAA) as a key metabolite of Trp modulating vascular inflammation and lipid metabolism. The molecular mechanisms driven by 3-HAA in atherosclerosis have not been completely elucidated. In this study, we investigated whether two major signalling pathways, activation of SREBPs and inflammasome, are associated with the 3-HAA-dependent regulation of lipoprotein synthesis and inflammation in the atherogenesis process. Moreover, we examined whether inhibition of endogenous 3-HAA degradation affects hyperlipidaemia and plaque formation. In vitro, we showed that 3-HAA reduces SREBP-2 expression and nuclear translocation and apolipoprotein B secretion in HepG2 cell cultures, and inhibits inflammasome activation and IL-1β production by macrophages. Using Ldlr-/- mice, we showed that inhibition of 3-HAA 3,4-dioxygenase (HAAO), which increases the endogenous levels of 3-HAA, decreases plasma lipids and atherosclerosis. Notably, HAAO inhibition led to decreased hepatic SREBP-2 mRNA levels and lipid accumulation, and improved liver pathology scores. We show that the activity of SREBP-2 and the inflammasome can be regulated by 3-HAA metabolism. Moreover, our study highlights that targeting HAAO is a promising strategy to prevent and treat hypercholesterolaemia and atherosclerosis.


Intracerebroventricular administration of cobalt protoporphyrin elicits prolonged weight reduction in rats.
Intracerebroventricular administration of the synthetic heme analogue cobalt protoporphyrin (CoPP) results in a dose-dependent decrease in body weight in rats. Among several other metalloporphyrins tested, this effect was found to be specific to cobalt chelated in native or minimally modified protoporphyrin rings. Metabolic balance studies showed that the initial effect of intracerebroventricular treatment with CoPP (0.4 mumol/kg body wt) was a transient reduction in food intake. However, following resumption of normal food intake and growth rate, absolute body weights remained reduced for prolonged periods of time (200-300 days) in both male and female rats. Heme oxygenase activity was induced and cytochrome P-450 activities were reduced in both brain and hypothalamus following intracerebroventricular administration of CoPP. The proximate mechanism of action of this synthetic heme analogue is not known at present, but it appears to act in the central nervous system, probably in the vicinity of the hypothalamus, to reduce the body weight set point of treated animals.


Effect of estimated glomerular filtration rate on periprocedural myocardial infarction in patients undergoing elective percutaneous coronary intervention.
Little is known about the effect of the estimated glomerular filtration rate (eGFR) on the periprocedural myocardial infarction (PMI). The aim of this study was to determine an eGFR value that is related with PMI development in patients with stable angina undergoing elective percutaneous coronary intervention (PCI). A retrospective analysis was conducted of 257 consecutive PCI patients with stable angina pectoris. The patients were divided into three groups according to eGFR: Group 1: eGFR > 90 mL/min/1.73 m(2), Group 2: eGFR = 60-89 mL/min/1.73 m(2), and Group 3: eGFR = 30-59 mL/min/1.73 m(2). Cardiac biomarkers were measured before, at 8, and at 24 h after the procedure. Periprocedural myocardial infarction occurred in 19% of the study patients. The frequency of PMI was 13.8% in group 1, 15.2% in group 2, and 35% in group 3 (p = 0.002). There was an inverse relationship with increasing cardiac biomarkers and decreasing eGFR values. Multiple regression analysis showed that an eGFR value between 30 and 59 mL/min/1.73 m(2) was an independent variable that significantly affected PMI development after PCI. An estimated glomerular filtration rate between 30 and 59 mL/min/1.73 m(2) is a predictor of developing PMI after elective PCI in patients with stable angina pectoris.


Intrinsic membrane association of the cytoplasmic tail of influenza virus M2 protein and lateral membrane sorting regulated by cholesterol binding and palmitoylation.
The influenza virus transmembrane protein M2 is a proton channel, but also plays a role in the scission of nascent virus particles from the plasma membrane. An amphiphilic helix in the CT (cytoplasmic tail) of M2 is supposed to insert into the lipid bilayer, thereby inducing curvature. Palmitoylation of the helix and binding to cholesterol via putative CRAC (cholesterol recognition/interaction amino acid consensus) motifs are believed to target M2 to the edge of rafts, the viral-budding site. In the present study, we tested pre-conditions of this model, i.e. that the CT interacts with membranes, and that acylation and cholesterol binding affect targeting of M2. M2-CT, purified as a glutathione transferase fusion protein, associated with [3H]photocholesterol and with liposomes. Mutation of tyrosine residues in the CRAC motifs prevented [(3)H]photocholesterol labelling and reduced liposome binding. M2-CT fused to the yellow fluorescent protein localized to the Golgi in transfected cells; membrane targeting was dependent on CRAC and (to a lesser extent) on palmitoylation. Preparation of giant plasma membrane vesicles from cells expressing full-length M2-GFP (green fluorescent protein) showed that the protein is partly present in the raft domain. Raft targeting required palmitoylation, but not the CRAC motifs. Thus palmitoylation and cholesterol binding differentially affect the intrinsic membrane binding of the amphiphilic helix.


Application of structural analysis to the mechanical behaviour of the cornea.
Structural engineering analysis tools have been used to improve the understanding of the biomechanical behaviour of the cornea. The research is a multi-disciplinary collaboration between structural engineers, mathematical and numerical analysts, ophthalmologists and clinicians. Mathematical shell analysis and nonlinear finite-element modelling have been used in conjunction with laboratory experiments to study the behaviour of the cornea under different loading states and to provide improved predictions of the mechanical response to disease and injury. The initial study involved laboratory tests and mathematical back analysis to determine the corneal material properties and topography. These data were then used to facilitate the construction of accurate finite-element models that are able to reliably trace the performance of cornea upon exposure to disease, injury or elevated intra-ocular pressure. The models are being adapted to study the response to keratoconus (a disease causing loss of corneal tissue) and to tonometry procedures, which are used to measure the intra-ocular pressure. This paper introduces these efforts as examples of the application of structural engineering analysis tools and shows their potential in the field of corneal biomechanics.


Drought and child mortality: a meta-analysis of small-scale surveys from Ethiopia.
Despite the intuitive connection between drought and mortality, we still lack a sound quantitative synthesis of evidence drawn from the available data. In this study, we estimate the pooled under-five death rates (U5DR) and assess the effect of drought on child death in Ethiopia. Small-scale mortality surveys were searched from the Complex Emergency Database and then aggregated spatially and temporally with drought exposure data from the Global Drought Monitor and food insecurity data from the Famine Early Warning Systems Network. A Bayesian Poisson meta-analysis was performed on 88 surveys conducted in Ethiopia between 2009 and 2014, consisting of 55,219 under-five children. The pooled U5DR was estimated at 0.323/10,000/day (95% credible interval, CrI: 0.254-0.397), which is below both the emergency and the baseline death rate thresholds of sub-Saharan Africa. We failed to find a plausible association between drought and U5DR. However, minimal food insecure areas showed elevated U5DR compared to stressed food insecure areas. Furthermore, the U5DR increases as the prevalence of acute malnutrition increases. Targeted interventions to improve the underlying causes of child malnutrition are crucial. Further, revising and updating the existing mortality thresholds, both the baseline and the emergency, is recommended.


Bayesian methods: a potential path forward for sepsis trials.
Given the success of recent platform trials for COVID-19, Bayesian statistical methods have become an option for complex, heterogenous syndromes like sepsis. However, study design will require careful consideration of how statistical power varies using Bayesian methods across different choices for how historical data are incorporated through a prior distribution and how the analysis is ultimately conducted. Our objective with the current analysis is to assess how different uses of historical data through a prior distribution, and type of analysis influence results of a proposed trial that will be analyzed using Bayesian statistical methods. We conducted a simulation study incorporating historical data from a published multicenter, randomized clinical trial in the US and Canada of polymyxin B hemadsorption for treatment of endotoxemic septic shock. Historical data come from a 179-patient subgroup of the previous trial of adult critically ill patients with septic shock, multiple organ failure and an endotoxin activity of 0.60-0.89. The trial intervention consisted of two polymyxin B hemoadsorption treatments (2 h each) completed within 24 h of enrollment. In our simulations for a new trial of 150 patients, a range of hypothetical results were observed. Across a range of baseline risks and treatment effects and four ways of including historical data, we demonstrate an increase in power with the use of clinically defensible incorporation of historical data. In one possible trial result, for example, with an observed reduction in risk of mortality from 44 to 37%, the probability of benefit is 96% with a fixed weight of 75% on prior data and 90% with a commensurate (adaptive-weighting) prior; the same data give an 80% probability of benefit if historical data are ignored. Using Bayesian methods and a biologically justifiable use of historical data in a prior distribution yields a study design with higher power than a conventional design that ignores relevant historical data. Bayesian methods may be a viable option for trials in critical care medicine where beneficial treatments have been elusive.