Assessment of the plasmidome of an extremophilic microbial community from the Diamante Lake, Argentina.
Diamante Lake located at 4589 m.a.s.l. in the Andean Puna constitutes an extreme environment. It is exposed to multiple extreme conditions such as an unusually high concentration of arsenic (over 300 mg L-1) and low oxygen pressure. Microorganisms thriving in the lake display specific genotypes that facilitate survival, which include at least a multitude of plasmid-encoded resistance traits. Hence, the genetic information provided by the plasmids essentially contributes to understand adaptation to different stressors. Though plasmids from cultivable organisms have already been analyzed to the sequence level, the impact of the entire plasmid-borne genetic information on such microbial ecosystem is not known. This study aims at assessing the plasmidome from Diamante Lake, which facilitates the identification of potential hosts and prediction of gene functions as well as the ecological impact of mobile genetic elements. The deep-sequencing analysis revealed a large fraction of previously unknown DNA sequences of which the majority encoded putative proteins of unknown function. Remarkably, functions related to the oxidative stress response, DNA repair, as well as arsenic- and antibiotic resistances were annotated. Additionally, all necessary capacities related to plasmid replication, mobilization and maintenance were detected. Sequences characteristic for megaplasmids and other already known plasmid-associated genes were identified as well. The study highlights the potential of the deep-sequencing approach specifically targeting plasmid populations as it allows to evaluate the ecological impact of plasmids from (cultivable and non-cultivable) microorganisms, thereby contributing to the understanding of the distribution of resistance factors within an extremophilic microbial community.


TGF-β1 modulates temozolomide resistance in glioblastoma via altered microRNA processing and elevated MGMT.
Our previous studies have indicated that miR-198 reduces cellular methylguanine DNA methyltransferase (MGMT) levels to enhance temozolomide sensitivity. Transforming growth factor beta 1 (TGF-β1) switches off miR-198 expression by repressing K-homology splicing regulatory protein (KSRP) expression in epidermal keratinocytes. However, the underlying role of TGF-β1 in temozolomide resistance has remained unknown. The distribution of KSRP was detected by western blotting and immunofluorescence. Microarray analysis was used to compare the levels of long noncoding RNAs (lncRNAs) between TGF-β1-treated and untreated cells. RNA immunoprecipitation was performed to verify the relationship between RNAs and KSRP. Flow cytometry and orthotopic and subcutaneous xenograft tumor models were used to determine the function of TGF-β1 in temozolomide resistance. Overexpression of TGF-β1 contributed to temozolomide resistance in MGMT promoter hypomethylated glioblastoma cells in vitro and in vivo. TGF-β1 treatment reduced cellular MGMT levels through suppressing the expression of miR-198. However, TGF-β1 upregulation did not affect KSRP expression in glioma cells. We identified and characterized 2 lncRNAs (H19 and HOXD-AS2) that were upregulated by TGF-β1 through Smad signaling. H19 and HOXD-AS2 exhibited competitive binding to KSRP and prevented KSRP from binding to primary miR-198, thus decreasing miR-198 expression. HOXD-AS2 or H19 upregulation strongly promoted temozolomide resistance and MGMT expression. Moreover, KSRP depletion abrogated the effects of TGF-β1 and lncRNAs on miR-198 and MGMT. Finally, we found that patients with low levels of TGF-β1 or lncRNA expression benefited from temozolomide therapy. Our results reveal an underlying mechanism by which TGF-β1 confers temozolomide resistance. Furthermore, our findings suggest that a novel combination of temozolomide with a TGF-β inhibitor may serve as an effective therapy for glioblastomas.


Evidence of calmodulin involvement in cell volume recovery following hypo-osmotic stress.
An influx of Ca2+ into red blood cells of the bivalve mollusc Noetia ponderosa occurs immediately following a hypo-osmotic stress. The volume recovery response to the stress is dependent upon [Ca2+]o and is inhibited by phenothiazines. The action of these drugs is on the amino acid regulation portion of the recovery rather than on the ionic portion. Since the phenothiazines are non-specific in action, we have conducted several experiments to decide the site of phenothiazine action on the volume recovery response. The sulfoxide derivatives of both chlorpromazine and trifluoperazine have no effect on volume regulation at the same dose where the parent compound inhibits. At 50-100 times the concentration of the parent compound, the derivatives block both volume regulation and taurine efflux. The phorbol ester, TPA, an activator of protein kinase C, alters the volume recovery, but does so by affecting K+ rather than amino acid regulation. The only phenothiazine target that we can not rule out is calmodulin, which we also demonstrate to be present in the clam red cells. Thus, the data presented suggest that calmodulin is involved in the amino acid regulatory portions of the volume recovery in response to hypo-osmotic swelling.


Impact of intramolecular hydrogen bonding of gallic acid conformers on chemical shift through NMR spectroscopy.
Intramolecular hydrogen bonding of gallic acid conformers was probed as a function of their dihedral angles using 13C nuclear magnetic resonance (NMR) chemical shift (δC). The quantum mechanically calculated 13C NMR chemical shift based on the most stable conformer (GA-I) in dimethylsulfoxide (DMSO) solution agrees to available measurement in the same solvent (RMSD = 0.95 ppm), better than to the measurement in solid phase (RMSD = 1.93 ppm). The accuracy of the calculated NMR chemical shift of the nominal but non-equivalent phenyl carbons C(3)/C(7) and C(4)/C(6) (ortho and meta to the acid -C(1)OOH group) of GA may not be evaluated using the experimental measurements at room temperature. The splitting in chemical shift of the nominal phenyl carbons is able to be experimentally measured only in low temperature NMR and using quantum mechanical calculations. We further recognised that the C NMR chemical shifts of the nominal phenyl carbons (C(3)/C(7) and C(4)/C(6)) encode information for intramolecular hydrogen bonding network formed by GA conformers. The ability to obtain accurate splitting of NMR chemical shifts for nominal carbons, therefore, determines the usefulness of the NMR technique as a probe for conformation of GA.


A mixed methods approach to identifying factors related to voluntary HIV testing among injection drug users in Shanghai, China.
Injection drug use is a major route of HIV transmission in China, yet relatively little is known about why so few injection drug users utilize free HIV testing services. This study aimed to examine barriers to HIV testing and voluntary counseling and testing (VCT) service utilization among injection drug users in Shanghai, China. Utilizing mixed methods, we analyzed data from a survey of 540 compulsory drug abuse treatment patients and data from focus groups with 70 service providers and patients. Only 24.4% of patients expressed willingness to be tested for HIV. Willingness to be tested was associated with younger age and more positive attitudes towards condom use. Patients reported several barriers to utilization of voluntary HIV testing services, including lack of information about these services, perceptions of no risk or low-risk for HIV infection, fear of positive results, and the stigma or discrimination that may be experienced by the patient or their family. Having limited skills related to HIV counseling was reported by service providers as the primary barrier to encouraging patients to utilize HIV testing/VCT services. Special intervention programs targeting injection drug users, their family members, and service providers may increase HIV testing in China.


Deep molecular divergence and exceptional morphological stasis in dwarf cannibal snails Nata sensu lato Watson, 1934 (Rhytididae) of southern Africa.
The genus Nata Watson, 1934 is a southern African endemic belonging to the Gondwanan family of carnivorous snails, Rhytididae. We present a molecular phylogeny of the genus based on two mitochondrial (16S and COI) and two nuclear genes (ITS2 and 28S RNA), and complement this with an appraisal of morphological characters relating to both the shell and soft parts. We identify four reciprocally monophyletic lineages for which valid names are already available, plus two undescribed species restricted to the Albany Thicket Biome. We show that Nata sensu lato may not be monophyletic. Rather there exist two deep lineages within Nata s.l., one lineage potentially sister to a clade dominated by the Australian and New Zealand radiation, and the other occupying a basal position within Rhytididae. Accordingly we recommend a revision recognising two genera, namely Nata s.s. and Natella respectively. Despite deep molecular divergences within Nata s.s., phenotypic evolution has been remarkably conserved, and contrasts greatly with that exhibited across other major lineages within the Rhytididae.


Characterization of mucosa-associated Escherichia coli strains isolated from Crohn's disease patients in Brazil.
Crohn's disease (CD) is characterized by chronic inflammation of the human intestine. Several studies have demonstrated that the intestinal mucosa of CD patients in Western countries is abnormally colonized by adherent-invasive Escherichia coli (AIEC) strains. However, no studies to date have focused on the involvement of such E. coli strains in CD patients in Brazil. Here, we characterized E. coli strains associated with the ileal mucosa of Brazilian CD patients (ileal biopsies from 35 subjects, 24 CD patients and 11 controls). The colonization level of adherent Enterobacteriaceae associated with the ileal mucosa of CD patients was significantly higher than that of the controls. The proportions of E. coli strains belonging to phylogroups B1 and B2 were two-fold higher in strains isolated from CD patients than in those isolated from controls. CD patients in the active phase harbored 10-fold more E. coli belonging to group B2 than CD patients in remission. Only a few E. coli isolates had invasive properties and the ability to survive within macrophages, but 25% of CD patients in Brazil (6/24) harbored at least one E. coli strain belonging to the AIEC pathobiont. However, fimH sequence analysis showed only a few polymorphisms in the FimH adhesin of strains isolated in this study compared to the FimH adhesin of AIEC collections isolated from European patients. Mucosa-associated E. coli strains colonize the intestinal mucosa of Brazilian CD patients. However, the strains isolated from Brazilian CD patients have probably not yet co-evolved with their hosts and therefore have not fully developed a strong adherent-invasive phenotype. Thus, it will be crucial to follow in the future the emergence and evolution of AIEC pathobionts in the Brazilian population.


Replacing microglia to treat Alzheimer's disease.
In this issue of Cell Stem Cell, Yoo et al.1 report that replacing Trem2 knockout microglia with Trem2 wild-type microglia-like cells derived from systemically transplanted hematopoietic cells rectifies microglial dysfunction in a mouse model of amyloidosis. These findings highlight the potential of microglia replacement therapy for Alzheimer's disease treatment.


Polyamines modulate streptomycin-induced mistranslation in Escherichia coli.
The effects of intracellular levels of polyamines on both the in vivo inhibition of protein synthesis and the decrease of translation accuracy induced by streptomycin have been studied in polyamine-auxotrophic strains of Escherichia coli infected with the MS2 bacteriophage. The amount of viral coat protein formed was strongly reduced upon addition of increasing concentrations of streptomycin to polyamine-supplemented bacteria. In contrast, the antibiotic almost did not inhibit coat protein synthesis in polyamine-starved cells. The increase of mistranslation frequency elicited by streptomycin was only observed in bacteria grown with putrescine. In these cells several coat protein-satellites were detected after two-dimensional gel electrophoresis. These proteins, more basic than the normal MS2 coat protein, contain multiple substitutions of lysine for asparagine.


Re-alignment procedure for ulnar drift of the metacarpophalangeal joint in rheumatoid arthritis.
An operation for correcting ulnar deviation in rheumatoid arthritis, by employing local tendon transfers at an early stage in the developing deformity, is described. The pathology, clinical staging and results in 103 joints are presented.


Compatible osmolytes modulate the response of porcine endothelial cells to hypertonicity and protect them from apoptosis.
Porcine pulmonary arterial endothelial cells accumulated myo-inositol and taurine, as well as betaine, during adaptation to hypertonic stress. The cells grew and maintained their normal morphology during culture in hypertonic (0.5 osmol (kg H(2)O)(-1)) medium that contained osmolytes such as betaine, myo-inositol or taurine at concentrations close to reported physiological values. The cells did not grow well in hypertonic medium depleted of potential compatible osmolytes. After a few days, cell density decreased by about 50 % and many cells rounded up and detached from the plates, their nuclei showing clear apoptotic morphology. The caspase-3 activity of the cells also increased dramatically under these conditions, but remained negligibly low when betaine and myo-inositol were added to the medium. Addition of betaine and myo-inositol to hypertonic medium depleted of compatible osmolytes increased the number of colonies remaining after 12 days of culture; with each solute at 30-100 micromol l(-1) the number increased about sixfold. In the absence of compatible osmolytes, increased mRNA levels and corresponding activities of betaine/gamma-aminobutyric acid transporter (BGT1) and sodium/myo-inositol transporter (SMIT) induced by hypertonicity remained high after 72 h incubation, whereas they were down regulated in the presence of betaine and myo-inositol. Similarly, the down regulation of the amino acid System A transporter (ATA2) was markedly slowed in the absence of compatible osmolytes. We conclude that these compatible osmolytes at concentrations close to physiological values enable the endothelial cells to adapt to hypertonic stress, protecting them from apoptosis, and also modulate the adaptation process.


Multimodality imaging review of breast vascular lesions.
Breast vasculature is visualised on all screening and diagnostic imaging of the breast. Various vascular breast lesions exist, spanning from chronic systemic processes, congenital malformations, post-traumatic sequela, benign masses, to malignant tumours. Accurate diagnosis of vascular lesions on breast imaging can be difficult due to overlapping characteristics. Radiologists should be aware of key multi-technique imaging features to make an accurate diagnosis and to avoid unnecessary biopsies. This article reviews the normal vascular anatomy of the breast, commonly used imaging techniques to diagnose vascular lesions, and an in-depth review of various vascular breast lesions. Finally, management recommendations and clinical guidance are discussed so that the radiologist can appropriately triage these patients.


Predictors of stress among parents in pediatric intensive care unit: a prospective observational study.
To determine the sociodemographic and clinical factors leading to stress among parents whose children are admitted in pediatric intensive care unit (PICU). A prospective observational study was conducted in PICU of a tertiary care hospital of north India. Parents of children admitted to PICU for at least 48 h duration were eligible for participation. At the end of 48 h, parental stress was assessed using parental stress scale (PSS:PICU) questionnaire which was administered to the parents. Baseline demographic and clinical parameters of children admitted to PICU were recorded. The parental stress was compared with demographic and clinical characteristics of children using appropriate statistical methods. A total of 49 parents were finally eligible for participation. Mean (SD) parental stress scores was highest in domains of procedures [1.52 (0.66)] and behavior and emotional [1.32 (0.42)] subscales. Mean (SD) total parental stress score among intubated children [1.31 (0.25)] was significantly more than among non intubated children [0.97 (0.26)] (p < 0.001). However, parental stress score were comparable in terms of gender (p = 0.15) and socioeconomic status (p = 0.32). On subscale analysis, it was found that professional communication is a significant stressor in age groups 0-12 mo [0.61(0.41)] (p = 0.02). It was observed that parents of intubated children were significantly stressed by the physical appearance of their children (p < 0.001), procedures performed on them (p = 0.008) and impairment in parental role (p = 0.002). Total parental stress score had a positive correlation with PRISM score (r = 0.308). Indian parents are stressed maximally with environment of PICU. Factor leading to parental stress was intubation status of the child and was not affected by gender or socio demographic profile of the parents.


Internalizing cancer antibodies from phage libraries selected on tumor cells and yeast-displayed tumor antigens.
A number of approaches have been utilized to generate antibodies to cancer cell surface receptors that can be used as potential therapeutics. A number of these therapeutic approaches, including antibody-drug conjugates, immunotoxins, and targeted nucleic acid delivery, require antibodies that not only bind receptor but also undergo internalization into the cell upon binding. We previously reported on the ability to generate cancer cell binding and internalizing antibodies directly from human phage antibody libraries selected for internalization into cancer cell lines. While a number of useful antibodies have been generated using this approach, limitations include the inability to direct the selections to specific antigens and to identify the antigen bound by the antibodies. Here we show that these limitations can be overcome by using yeast-displayed antigens known to be associated with a cell type to select the phage antibody output after several rounds of selection on a mammalian cell line. We used this approach to generate several human phage antibodies to yeast-displayed EphA2 and CD44. The antibodies bound both yeast-displayed and mammalian cell surface antigens, and were endocytosed upon binding to mammalian cells. This approach is generalizable to many mammalian cell surface proteins, results in the generation of functional internalizing antibodies, and does not require antigen expression and purification for antibody generation.


Ablation of free radical-mediated reperfusion injury for the salvage of kidneys taken from non-heartbeating donors. A quantitative evaluation of the proportion of injury caused by reperfusion following periods of warm, cold, and combined warm and cold ischemia.
Postischemic renal failure is a severe problem following cadaveric renal transplantation, especially if the kidney has been harvested from a non-heartbeating donor, and thereby subjected to periods of both warm and cold ischemia. It is well established that a substantial component of postischemic injury is produced by oxygen-derived free radicals generated from xanthine oxidase at reperfusion. However, the clinical potential of free radical ablative therapy is dependent upon the proportion of the total injury caused by this reperfusion mechanism, compared with the proportion resulting from ischemic injury per se. Therefore, we quantitatively evaluated these proportions in porcine kidneys subjected to various periods of warm (renal artery occlusion in situ), cold (harvest, cold preservation, and allotransplantation), and combined warm and cold ischemia. Experiments were paired, one kidney treated with either superoxide dismutase (SOD) or allopurinol for free radical ablation, the contralateral kidney serving as a control. Creatinine clearance (Ccr) was measured separately for each kidney 48 hr after reperfusion. After 1 and 2 hr of warm ischemia, Ccr dropped to 50% and 36% of normal, respectively. This was improved to 110% and 55% when SOD was given into the renal artery at reperfusion. Similarly, after 24 and 48 hr of cold ischemia, kidney function was significantly improved from 30% and 18% to 72% and 47% of normal, respectively, when allopurinol was added to the preservation solution. SOD used at harvest and again at reperfusion was particularly effective following combined warm and cold ischemia, in a situation mimicking the harvest of cadaver kidneys from a non-heartbeating donor. These findings suggest that the ablation of free radical-mediated reperfusion injury may improve posttransplant renal function sufficiently to allow expansion of the cadaveric donor pool to include non-heartbeating donors.


Sevoflurane: an ideal agent for adult day-case anesthesia?
Sevoflurane has several properties which make it potentially useful as a day case anaesthetic. Following induction of anaesthesia with propofol, awakening from sevoflurane is faster compared to isoflurane, faster or similar compared to propofol and comparable (in the majority of studies) to desflurane. Subsequent recovery and discharge is generally similar following all agents. Sevoflurane may also be used to induce anaesthesia, which is generally well-received and causes less hypotension and apnoea compared to propofol. When used as a maintenance anaesthetic, the incidence of postoperative nausea and vomiting after sevoflurane is comparable to other inhaled anaesthetics, but this complication appears more common after inhaled inductions. The tolerability and low solubility of sevoflurane facilitate titration of anaesthesia and may reduce the need for opioid analgesia, which in turn may limit the occurrence of nausea and vomiting.


Polyoma virus and cyclic AMP-mediated control of dihydrofolate reductase mRNA abundance in methotrexate-resistant mouse fibroblasts.
As a model cell culture system for studying polyoma-mediated control of host gene expression, we isolated methotrexate-resistant 3T6 cells in which one of the virus-induced enzymes, dihydrofolate reductase, is a major cellular protein. In highly methotrexate-resistant cell lines dihydrofolate reductase synthesis accounts for over 10% that of soluble portein, corresponding to an increase of approximately 100-fold over the level in parental cells. This increase in dihydrofolate reductase synthesis is due to a corresponding increase in the abundance of dihydrofolate reductase mRNA and gene sequences. We have used these cells to show that infection with polyoma virus results in a 4- to 5-fold increase in the relative rate of dihydrofolate reductase synthesis and a corresponding increase in dihydrofolate reductase mRNA abundance. The increase in dihydrofolate reductase synthesis begins 15 to 20 h after infection and continues to increase until cell lysis. These observations represent the first direct evidence that viral infection of eukaryotic cells results in the increased synthesis of a specific cellular enzyme and an increase in the abundance of a specific cellular mRNA. In order to gain additional insight into the control of dihydrofolate reductase synthesis we examined other parameters affecting dihydrofolate reductase synthesis. We found that the addition of fresh serum to stationary phase cells results in a 2-fold stimulation of dihydrofolate reductase synthesis, beginning 10 to 12 h after serum addition. Serum stimulation of dihydrofolate reductase synthesis is completely inhibited by the presence of dibutyryl cyclic AMP as well as by theophylline or prostaglandin E1, compounds which cause an increase in intracellular cyclic AMP levels. In fact, the presence of dibutyryl cyclic AMP and theophylline results in a 2- to 3-fold decrease in the rate of dihydrofolate reductase synthesis and the abundance of dihydrofolate reductase mRNA. However, in contrast to the effect on serum stimulation, dibutyryl cyclic AMP and theophylline do not inhibit polyoma virus induction of dihydrofolate reductase synthesis or dihydrofolate reductase mRNA levels. These observations suggest that dihydrofolate reductase gene expression is controlled by at least two regulatory pathways: one involving serum that is blocked by high levels of cyclic AMP and another involving polyoma induction that is not inhibited by cyclic AMP.


Comparison of the performance of the borax buffer-based HRP-enhanced reagent and the 'Lumi-Phos 530' chemiluminescence systems in the detection of biotinylated DNA.
A comparison of two chemiluminescence methods, the borax buffer-based HRP-enhanced reagent and Lumi-Phos 530, applied to the detection of a biotinylated 30-mer DNA slot blotted onto a nylon membrane, is presented. A streptavidin-HRP and streptavidin-ALP mediated detection system was used. The HRP-enhanced system is up to 15-fold greater with respect to the signal/background ratios than the Lumi-Phos 530 system at 0.5 microgram biotinylated DNA with at least a two-fold improvement in detection sensitivity for 0.5 ng biotinylated DNA.


Virus-derived gene transfer agents benefit host cells by providing templates for DNA repair.
The potential benefits of producing gene transfer agents (GTAs) have long been speculated. A new study in PLOS Biology shows that DNA transfer by these phage-like elements allows cells with DNA damage to perform DNA repair and survive.


Electrophysiology and antiarrhythmic efficacy of intravenous pirmenol in patients with sustained ventricular tachyarrhythmias.
We assessed the electrophysiologic effects and antiarrhythmic efficacy of intravenous pirmenol in 15 patients who had spontaneous and induced sustained ventricular tachyarrhythmias. At a plasma concentration of 2.29 +/- 0.75 micrograms/ml, pirmenol decreased sinus cycle length by 11 +/- 13%, increased QRS, QTc, and HV intervals by 14 +/- 12%, 13 +/- 12%, and 22 +/- 28%, respectively, and increased atrial and ventricular effective refractory periods (ERP) by 20 +/- 14% and 7 +/- 8%, respectively. There was a greater increase in QRS duration during ventricular tachycardia and ventricular pacing than during sinus rhythm (p less than 0.005). By electropharmacologic testing, pirmenol was judged effective in six patients (40%) and was proarrhythmic in one (6%). In the nine patients in whom pirmenol was judged ineffective, the cycle length of induced VT increased by 36 +/- 15% and the associated mean arterial pressure increased by 21 +/- 14 mm Hg. The only side effects were mild hypotension and mild nausea in one patient each. Intravenous pirmenol has type IA electrophysiologic effects. It can be administered safely to patients with sustained ventricular tachyarrhythmias and is as effective as approved antiarrhythmic drugs when assessed by electropharmacologic testing.


Ultrafast isolated molecule imaging without crystallization.
Crystallography is the standard for determining the atomic structure of molecules. Unfortunately, many interesting molecules, including an extensive array of biological macromolecules, do not form crystals. While ultrashort and intense X-ray pulses from free-electron lasers are promising for imaging single isolated molecules with the so-called "diffraction before destruction" technique, nanocrystals are still needed for producing sufficient scattering signal for structure retrieval as implemented in serial femtosecond crystallography. Here, we show that a femtosecond laser pulse train may be used to align an ensemble of isolated molecules to a high level transiently, such that the diffraction pattern from the highly aligned molecules resembles that of a single molecule, allowing one to retrieve its atomic structure with a coherent diffraction imaging technique. In our experiment with CO2 molecules, a high degree of alignment is maintained for about 100 fs, and a precisely timed ultrashort relativistic electron beam from a table-top instrument is used to record the diffraction pattern within that duration. The diffraction pattern is further used to reconstruct the distribution of CO2 molecules with atomic resolution. Our results mark a significant step toward imaging noncrystallized molecules with atomic resolution and open opportunities in the study and control of dynamics in the molecular frame that provide information inaccessible with randomly oriented molecules.


The antinociceptive effects of intrathecally administered levonantradol and desacetyllevonantradol in the rat.
Levonantradol and its desacetylated metabolite (desacetyllevonantradol) produced a dose-dependent increase in the hot plate and tail flick response latencies following intrathecal administration in a dose range from 4 to 40 micrograms. No difference in potency between the two drugs was observed, as defined by the ED50 values obtained in either test. The duration of the effect of either drug was also dose dependent ranging from 30 to 120 minutes. No effect on placing, stepping or righting reflexes or the ability of the rats to negotiate a 60 degrees inclined plane was observed at these doses. At the highest doses, however, an exaggerated myoclonic response to abrupt muscle stretch was observed. None of those effects was antagonized by naloxone (2 mg/kg) administered intraperitoneally before or after levonantradol.


[Acceptance of fecal microbiota transfer among patients with chronic inflammatory bowel diseases in a highly specialized outpatient department: a questionnaire-based survey].
Recently, research in the treatment of inflammatory bowel diseases has become increasingly focused on fecal microbiota transfer (FMT) due to increasing evidence of its possible benefits. Still, there are doubts about this method, because there is contradicting evidence regarding its effectiveness and the possible side effects are not well known. Furthermore, the majority of patients are not open to this procedure. We performed a questionnaire-based survey amongst 302 patients with an inflammatory bowel disease that received treatment in our specialized outpatient clinic to determine the factors relevant for acceptance or rejection of fecal microbiota transfer as a possible treatment for Crohn's disease or ulcerative colitis. Our data supports the hypothesis that a lack of information about FMT is a key factor for hypothetical acceptance of this method (68 % of pre-informed participants vs. 30 % of not pre-informed participants would accept FMT as treatment, p < 0.001), and, therefore, it highlights patient education as a possible intervention to improve acceptance. The main concern regarding FMT was possible transmission of infections (ranked first by 98 participants). The most accepted method to perform FMT was application via oral capsule (44 % of participants).


DL-3-n-butylphthalide improves the endothelium-dependent vasodilation in high-fat diet-fed ApoE-/- mice via suppressing inflammation, endothelial necroptosis and apoptosis.
Impaired endothelium-dependent vasodilation in atherosclerosis is a high-risk factor for myocardial infarction and ischemic stroke, and inflammation, necroptosis and apoptosis contribute to endothelial dysfunction in atherosclerosis. Although DL-3-n-butylphthalide (NBP) has been widely used in treating ischemic stroke, its effect on endothelium-dependent vasodilation remains unknown. This study aims to explore whether NBP is able to improve endothelium-dependent vasodilation in atherosclerosis and the underlying mechanisms. Male ApoE-/- mice were fed with a high-fat diet (HFD) for 9-16 weeks to establish a model of atherosclerosis. NBP were given to the mice after eating HFD for 6 weeks and atorvastatin served as a positive control. The endothelium-dependent vasodilation, the blood flow velocity, the atherosclerotic lesion area, the serum levels of lipids, inflammatory cytokines and necroptosis-relevant proteins (RIPK1, RIPK3 and MLKL), and the endothelial necroptosis and apoptosis within the aorta were measured. Human umbilical vein endothelial cells (HUVECs) were incubated with oxidized low-density lipoprotein (ox-LDL) for 48 h to mimic endothelial injury in atherosclerosis, lactate dehydrogenase release, the ratio of necroptosis and apoptosis and the expression of necroptosis-relevant proteins were examined. Similar to atorvastatin, NBP improves endothelium-dependent vasodilation, decreases aortic flow velocity and reduces atherosclerotic lesion area in HFD-fed ApoE-/- mice, concomitant with a reduction in serum lipids, inflammatory cytokines and necroptosis-relevant proteins, and endothelial necroptosis and apoptosis. Consistently, NBP inhibited necroptosis and apoptosis in ox-LDL-treated HUVECs. Based on these observations, we conclude that NBP exerts beneficial effects on improving the endothelium-dependent vasodilation in atherosclerosis via suppressing inflammation, endothelial necroptosis and apoptosis.


Occipital sharp waves in idiopathic partial epilepsies--clinical and genetic aspects.
In order to gain new insight into the pathogenesis and nosography of benign partial epilepsy with occipital paroxysms (BEOP) we retrospectively analysed cases with benign focal sharp waves (SHW) of different localisations, in which analogous EEG changes had been found in at least one first degree relative. Fifty-six families were evaluated. Nineteen cases with occipital SHW (group A) were compared with 37 cases without (group B). There was a broad spectrum of symptomatology with large overlaps between the two groups. However, a number of striking differences, yielding a characteristic picture of early childhood epilepsy with occipital foci were identified: (1) Age of onset under 5 years; (2) a larger proportion of febrile convulsions (FC 47 vs. 19%); (3) a trend towards a higher rate of typical early childhood occipital seizures (26 vs. 5%); (4) a higher rate of frontal and generalizing SHW foci (32 vs. 5%; 37 vs. 11%); (5) a higher rate of generalized spikes and waves (SW) (46 vs. 14%); (6) a trend towards a higher rate of photoparoxysmal response (PPR) (57 vs. 32%). The high prevalence of independent genetic traits favours a multifactorial pathogenesis. The predisposition to FC with characteristic early seizure onset and varying patterns of generalized genetic EEG traits plays a crucial role within the complex pathogenetic network. The early-onset benign childhood occipital seizure susceptibility syndrome of Panayiotopoulos (Benign Childhood Partial Seizures and Related Epileptic Syndromes, John Libbey & Company Ltd., London (1999)) cannot be regarded as the sole representative of occipital spikes in early childhood but as an important even though rare form of occipital epilepsy.


EoE Down Under Is Still EoE: Variability in Provider Practice Patterns in Australia and New Zealand Among Pediatric Gastroenterologists.
There is likely variation in approach and management of patient with EoE due to lack of standardized care and variation in guidelines. We aimed to identify current practices regarding diagnosis and treatment in children with eosinophillic esophagitis (EoE) in Australia and New Zealand (ANZ). Information on current diagnostic and management approaches for pediatric EoE was collected via an online survey sent to pediatric gastroenterologists (pGE) in ANZ. We performed a cross-sectional study of pGE using a 49-question instrument regarding evaluation, diagnostic, and therapeutic aspects of EoE between October 2019 and December 2019. Eighty-five percent of the survey responders were from Australia, and 66% were academic. 30% pGE perform > 3 esophageal biopsies for diagnosis of EoE, 40% involve an allergist, 30% use a twice daily PPI trial, and 70% do not exclude other cause of esophageal eosinophilia. For management, only 3% use dietary elimination as an initial therapy, and 24% use less than the recommended doses of swallowed fluticasone. Forty-nine percent were likely to stop treatment in after remission is achieved for 12 months. The EoE endoscopic reference score (EREFS) was not routinely used (49%). Two-thirds of pGE are concerned about long-term effects of recurrent need of general anesthesia. Diagnostic and management strategies for EoE differed widely among pGE in ANZ, including in diagnostic biopsies, assessing competing causes of esophageal eosinophilia, initials selection of treatments, and maintenance strategies. This variability likely reflects continued uncertainty regarding optimal management strategies and stresses the need for pediatric-specific ANZ guidelines to standardize EoE care.


Maternal molecular features and gene profiling of monocytes during first trimester pregnancy.
We examined the molecular characteristics of monocytes of pregnant and non-pregnant women to investigate the molecular effects that are associated with immunoregulation at the maternal-fetal interface. We analyzed molecular features and target genes in monocytes of pregnant women using flow cytometry, real-time PCR and oligonucleotide microarray technology. CD14(high) monocytes and several immune gene members including CD200, CD200R, IDO, IFI27, IL-10 and G0S2 were found to be differentially expressed in monocytes throughout pregnancy. In addition, transcripts within components of the signaling cascade of immune cells (HLA-DRB4, HBEGF, IL-8, CD3D, CCL5), and of several transcription factors (SOCS1, CXCL10, ID1, ID2) were altered in the monocytes of pregnant women. Further studies will be needed to elucidate the biological significance of our observation.


Arthroscopic Debridement and Microfracture of Osteochondral Lesion of the Talar Head.
Osteochondral lesions of the talar head can be classified into 4 types according to their location. A type 1 lesion is located at the anterior part of the talar head. Surgical debridement and microfracture are indicated for symptomatic type 1 lesions if conservative treatment fails to relieve the pain. The purpose of this technical note was to describe the details of arthroscopic debridement and microfracture of the symptomatic type 1 osteochondral lesion of the talar head and the kissing lesion at the navicular bone. The procedure is performed through the standard portals of talonavicular arthroscopy and has the advantages of minimally invasive surgery of better cosmetic results and less surgical trauma.


Peroperative three-dimensional correction of idiopathic scoliosis with the Cotrel-Dubousset procedure.
A peroperative measurement technique based on magnetic fields was used to document the thoracic and lumbar vertebral changes induced by derotation of the concave rod during corrective surgery of idiopathic scoliotic deformities with Cotrel-Dubousset instrumentation. Objective was to accurately document the immediate changes induced by the derotation maneuver to gain a better understanding of its effect on curve correction during a surgical procedure. Accurate peroperative documentation of these three-dimensional changes was very limited, and the exact contribution of the derotation maneuver in the entire process of correction of a scoliotic deformity was still poorly understood. A digitizer using magnetic fields was used to record the three-dimensional orientation and x, y, and z coordinates of the tip of every spinous process exposed at surgery before and after derotation of the concave rod in a group of 22 female patients with idiopathic scoliosis. A significant improvement of the scoliotic deformity was noted in the frontal plane, and improvement of thoracic hypokyphosis and lumbar hypolordosis in the sagittal plane was seen. Vertebral axial rotation remained unchanged. The derotation maneuver is effective in achieving three-dimensional correction of idiopathic scoliosis, but vertebral axial derotation is not an important component of this correction.


Enzymology of H2S biogenesis, decay and signaling.
Hydrogen sulfide (H2S), produced by the desulfuration of cysteine or homocysteine, functions as a signaling molecule in an array of physiological processes including regulation of vascular tone, the cellular stress response, apoptosis, and inflammation. The low steady-state levels of H2S in mammalian cells have been recently shown to reflect a balance between its synthesis and its clearance. The subversion of enzymes in the cytoplasmic trans-sulfuration pathway for producing H2S from cysteine and/or homocysteine versus producing cysteine from homocysteine, presents an interesting regulatory problem. It is not known under what conditions the enzymes operate in the canonical trans-sulfuration pathway and how their specificity is switched to catalyze the alternative H2S-producing reactions. Similarly, it is not known if and whether the mitochondrial enzymes, which oxidize sulfide and persulfide (or sulfane sulfur), are regulated to increase or decrease H2S or sulfane-sulfur pools. In this review, we focus on the enzymology of H2S homeostasis and discuss H2S-based signaling via persulfidation and thionitrous acid.


Glycemic Response to Corn Starch Modified with Cyclodextrin Glycosyltransferase and its Relationship to Physical Properties.
Corn starch was modified with cyclodextrin glycosyltransferase (CGTase) below the gelatinization temperature. The porous granules with or without CGTase hydrolysis products may be used as an alternative to modified corn starches in foods applications. The amount and type of hydrolysis products were determined, containing mainly β-cyclodextrin (CD), which will influence pasting behavior and glycemic response in mice. Irregular surface and small holes were observed by microscopic analysis and differences in pasting properties were observed in the presence of hydrolysis products. Postprandial blood glucose in mice fed gelatinized enzymatically modified starch peaked earlier than their ungelatinized counterparts. However, in ungelatinized enzymatically modified starches, the presence of β- CD may inhibit the orientation of amylases slowing hydrolysis, which may help to maintain lower blood glucose levels. Significant correlations were found between glycemic curves and viscosity pattern of starches.


Ferroptosis in plants: triggers, proposed mechanisms, and the role of iron in modulating cell death.
Regulated cell death plays key roles during essential processes throughout the plant life cycle. It takes part in specific developmental programs and maintains homeostasis of the organism in response to unfavorable environments. Ferroptosis is a recently discovered iron-dependent cell death pathway characterized by the accumulation of lipid reactive oxygen species. In plants, ferroptosis shares all the main hallmarks described in other systems. Those specific features include biochemical and morphological signatures that seem to be conserved among species. However, plant cells have specific metabolic pathways and a high degree of metabolic compartmentalization. Together with their particular morphology, these features add more complexity to the plant ferroptosis pathway. In this review, we summarize the most recent advances in elucidating the roles of ferroptosis in plants, focusing on specific triggers, the main players, and underlying pathways.


Factors associated with polypharmacy and the high risk of medication-related problems among older community-dwelling adults in European countries: a longitudinal study.
Polypharmacy can be defined as using five or more medications simultaneously. "Medication-related problems", an extension of polypharmacy, includes inappropriate prescribing, poor adherence, overdosage, underdosage, inappropriate drug selection, inadequate monitoring, adverse drug effects, and drug interactions. Polypharmacy and the high risk of medication-related problems among older people are associated with adverse health consequences due to drug-drug interactions, drug-disease interactions, and adverse drug effects. This study aims to assess the factors associated with polypharmacy and the high risk of medication-related problems among community-dwelling older people in the Netherlands, Greece, Croatia, Spain, United Kingdom. This longitudinal study used baseline and follow-up data from 1791 participants of the Urban Health Center European project. Polypharmacy and the risk of medication-related problems were evaluated at baseline and follow-up using the Medication Risk Questionnaire. We studied factors in the domains (a) sociodemographic characteristics, (b) lifestyle and nutrition, and (c) health and health care use. Hierarchical logistic regression analyses were used to examine the factors associated with polypharmacy and the high risk of medication-related problems. Mean age was 79.6 years (SD ± 5.6 years); 60.8% were women; 45.2% had polypharmacy, and 41.8% had a high risk of medication-related problems. Women participants had lower odds of polypharmacy (OR = 0.55;95%CI:0.42-0.72) and a high risk of medication-related problems (OR = 0.50; 95%CI:0.39-0.65). Participants with a migration background (OR = 1.67;95%CI:1.08-2.59), overweight (OR = 1.37; 95%CI:1.04-1.79) and obesity (OR = 1.78;95%CI:1.26-2.51) compared to 'normal weight', with lower physical HRQoL (OR = 0.96, 95%CI:0.95-0.98), multi-morbidity (OR = 3.73, 95%CI:2.18-6.37), frailty (OR = 1.69, 95%CI:1.24-2.30), visited outpatient services (OR = 1.77, 95%CI: 1.09-2.88) had higher odds of polypharmacy. The associations with the high risk of medication-related problems were similar. Multiple factors in demography, lifestyle, nutrition, and health care use are associated with polypharmacy and the high risk of medication-related problems. Polypharmacy is a single element that may reflect the number of medications taken. The broader content of medication-related problems should be considered to assess the context of medication use among older people comprehensively. These provide starting points to improve interventions to reduce polypharmacy and high risk of medication-related problems. In the meantime, health professionals can apply these insights to identify subgroups of patients at a high risk of polypharmacy and medication-related problems. The intervention of the UHCE project was registered in the ISRCTN registry as ISRCTN52788952. The date of registration is 13/03/2017.


Effect of non-homogeneous surface viscosity on the Marangoni migration of a droplet in viscous fluid.
Marangoni migration of a single droplet in an unbounded viscous fluid under the additional effect of variable surface viscosity is studied. The surface tension and the surface viscosity depend on concentration of dissolved species. Cases of the motion induced by the presence of a point source and by a given constant concentration gradient are considered. The dependence of the migration velocity on the governing parameters is computed under quasi-stationary approximation. The effect of weak advective transport is studied making use of singular perturbations in the Peclet number, Pe. It is shown that, when the source is time dependent a Basset-type history term appears in the expansion of the concentration and, as a result, the leading order correction to the flow and to the migration velocity is of O(Pe(1/2)). If the source of active substance driving the flow is steady, the effect of convective transport on the migration is weaker.


Key miRNAs and target genes played roles in the development of clear cell renal cell carcinoma.
Clear cell renal cell carcinoma (CCRCC) is the most aggressive form of renal cell carcinoma (RCC). This study was aimed to identify the differentially expressed miRNAs and target genes in CCRCC. The miRNA and mRNA next-generation sequencing data were downloaded from The Cancer Genome Atlas (TCGA) dataset. Differential expression analysis was performed, followed by correlation analysis of miRNA-mRNA. Functional enrichment and survival analysis was also performed. Seven hundred and eighty-seven patients with CCRCC from TCGA data portal were included in this study. A total of 52 differentially expressed miRNAs were identified in CCRCC. Then 2361 differentially expressed genes (DEGs) were identified. Prediction analysis and correlation analysis revealed that 89 miRNA-mRNA pairs were not only predicted by algorithms but also had a significant inverse relationship. Several differentially expressed miRNAs such as hsa-mir-501 and their target genes including AK1, SLC25A15 and PCDHGC3 had a significant prognostic value for CCRCC patients. Alterations of differentially expressed miRNAs and target genes may be involved in the development of CCRCC and can be considered as the prognostic markers for CCRCC.


Strain development and optimized fermentation conditions for blood meal using Aspergillus niger and Aspergillus oryzae.
To hydrolyze blood meal (BM) effectively, two mutants were generated using ultra-violet mutagenesis. Single-factor experiments, the Plackett-Burman technique and response surface methodology were used to optimize the fermentation conditions. This study successfully generated a mutant and innovatively provided important parameters for utilizing BM by fermentation, which could be of industrial value.


A comparison of four screening methods for cervical neoplasia.
To evaluate the sensitivity and accuracy of 4 screening methods for cervical neoplasia. Cervical samples from 450 women were evaluated for human papillomavirus DNA with a second-generation hybridization assay. Visual inspection, colposcopy, and liquid-based cytology were also performed for 273 of these women. The sensitivity, specificity, efficiency, positive predictive value (PPV), negative predictive value (NPV), Youden index, and kappa value of the different screening methods were compared. Liquid-based cytology was superior in specificity (98.63%) and PPV (92.86%) to visual inspection and colposcopy, while human papillomavirus DNA testing was superior in sensitivity (88.89%) and NPV (97.10%) to visual inspection and colposcopy. The best concordance with histologic findings was achieved by using both liquid-based cytology and viral DNA hybridization. Visual inspection and colposcopy should not be used when screening for early stage cervical lesions. The DNA hybridization assay is the best choice in primary screening, if available. Screening should begin at the age of 20 years.


Does staging computered tomography change management in thick malignant melanoma?
Histological confirmation and assessment of Breslow thickness are essential before embarking on the management plan in Malignant Melanoma (MM). Computerised Tomography (CT) is used in staging of MM in the UK according to BAD/BAPS (British Association of Dermatologists/British Association of Plastic Surgeons). Currently UK guidelines for the management of cutaneous melanoma at intermediate or high risk of recurrent disease (American Joint Cancer Committee) AJCC IIB disease or worse (Breslow 2.01-4.0mm with ulceration or Breslow >4mm) should have the following staging investigations: chest X-ray; liver ultrasonography or computed tomographic (CT) scan with intravenous contrast enhancement of chest, abdomen and pelvis; liver function tests; lactate dehydrogenase and full blood count. It has been the practice at our unit to perform a CT head and neck also as part of our staging. The aim of this study was to determine whether CT staging changed clinical management at the initial presentation scan and follow up scans. Also we aimed to see whether there was a benefit in performing CT head and neck in staging. A retrospective case note review was performed to see whether CT staging actually changed patient clinical management on 132 cases of AJCC IIB melanoma or worse over the past six years at our unit. Clinical management changes were divided into two groups: Initial presentation CT staging and follow up CT staging. In addition numbers of metastases to body regions were recorded. A total of 488 CT scans were performed on 132 patients (3.7 scans per patient). Initial presentation CT staging scans picked up 1/132 (0.7%) patient with an occult metastases that changed their clinical management. Of the 356 follow up CT staging scans imaging (11/127) 8.6% of patients had metastases detected and clinical management changed. All of these patients exhibited symptoms and signs of clinical metastatic disease. Head metastases are at least as common as other regions such as the chest & abdomen and more common than in the pelvis. Neck CT did not change management. CT staging for cutaneous melanoma is not indicated unless there are signs or symptoms of metastatic disease. If there are symptoms and signs of metastatic disease than patients should be staged and we advocate that staging of AJCC IIB/C should include imaging of the head in addition to chest, abdomen and pelvis.


Fecal immunochemical test (FIT) versus colonoscopy: Does knowing that a positive FIT requires a follow-up colonoscopy affect initial decision making in the US?
Although many people prefer fecal immunochemical test (FIT) over colonoscopy due to its noninvasive nature, it is unclear whether FIT would still be preferred for colorectal cancer (CRC) screening if they were explicitly informed that + FIT requires follow-up colonoscopy. To address this gap, we administered two conjoint analysis surveys-one that explained the need for a follow-up colonoscopy after + FIT while the other did not-to a US nationwide sample of Americans and then assessed whether there were differences in colonoscopy/FIT preferences between cohorts. We recruited adults ≥ 40yo who had not undergone CRC screening via an online survey research firm. We deployed two surveys that used conjoint analysis to assess decision making surrounding CRC screening tests: Survey 1 (4/2-4/15/2021)-did not inform participants that they need a colonoscopy following a + FIT; Survey 2 (4/29-6/2/2021)-informed respondents of the potential need. Using the conjoint analysis data, we determined the proportion of those who preferred FIT or colonoscopy and then used logistic regression to assess for differences in colonoscopy/FIT preferences between the cohorts. Overall, 501 and 1,000 individuals completed Survey 1 (without description of need for colonoscopy after + FIT) and Survey 2 (with description), respectively. There was no difference in test preference between cohorts after adjusting for covariates in the logistic regression (adjusted p = 0.09): Survey 1 (without description)-colonoscopy 28.5%, FIT 71.5%; Survey 2 (with description)-colonoscopy 26.7%, FIT 73.3%. Thus, knowledge that a + FIT requires a follow-up colonoscopy does not alter people's strong preference for non-invasive stool testing with FIT.


Allopurinol and oxypurinol differ in their strength and mechanisms of inhibition of xanthine oxidoreductase.
Xanthine oxidoreductase is a metalloenzyme that catalyzes the final steps in purine metabolism by converting hypoxanthine to xanthine and then uric acid. Allopurinol, an analog of hypoxanthine, is widely used as an antigout drug, as xanthine oxidoreductase-mediated metabolism of allopurinol to oxypurinol leads to oxypurinol rotation in the enzyme active site and reduction of the molybdenum Mo(VI) active center to Mo(IV), inhibiting subsequent urate production. However, when oxypurinol is administered directly to a mouse model of hyperuricemia, it yields a weaker urate-lowering effect than allopurinol. To better understand its mechanism of inhibition and inform patient dosing strategies, we performed kinetic and structural analyses of the inhibitory activity of oxypurinol. Our results demonstrated that oxypurinol was less effective than allopurinol both in vivo and in vitro. We show that upon reoxidation to Mo(VI), oxypurinol binding is greatly weakened, and reduction by xanthine, hypoxanthine, or allopurinol is required for reformation of the inhibitor-enzyme complex. In addition, we show oxypurinol only weakly inhibits the conversion of hypoxanthine to xanthine and is therefore unlikely to affect the feedback inhibition of de novo purine synthesis. Furthermore, we observed weak allosteric inhibition of purine nucleoside phosphorylase by oxypurinol which has potentially adverse effects for patients. Considering these results, we propose the single-dose method currently used to treat hyperuricemia can result in unnecessarily high levels of allopurinol. While the short half-life of allopurinol in blood suggests that oxypurinol is responsible for enzyme inhibition, we anticipate multiple, smaller doses of allopurinol would reduce the total allopurinol patient load.


Successful treatment of radiation-induced mucositis with proton pump inhibitor administration: A report of two laryngeal cancer cases.
Presently, the relationship between laryngopharyngeal reflux (LPR) and radiation-induced mucositis has not been fully explored. In the present study, we report 2 cases of laryngeal cancer in which radiation-induced mucositis ameliorated after proton pump inhibitor (PPI) administration. Case 1 was diagnosed with T1aN0M0 right glottis carcinoma and was treated with radiation therapy. Grade 3 mucositis occurred after administration of 46Gy irradiation. PPI was administered and mucositis ameliorated quickly without cessation of radiation therapy. Case 2 was diagnosed with T2N0M0 supraglottic cancer and was treated with concurrent chemoradiation therapy. Grade 3 mucositis occurred after administration of 44Gy irradiation. PPI was administered and mucositis ameliorated quickly without cessation of chemoradiation therapy. In both cases, a remarkable therapeutic effect of PPI was observed in the perilaryngeal areas including the epiglottic vallecula, arytenoid, and postcricoid area. In both cases, LPR involvement was suspected before the onset of radiation therapy. The two cases presented here, indicated a causal relationship between LPR and radiation-induced mucositis. In cases of severe mucositis in the perilaryngeal area in patients with LPR prior to radiation therapy, PPI administration may be an effective therapeutic option.


Soluble extracts from larval Ostertagia ostertagi modulating immune function.
BALB/c mice were immunized with Ostertagia ostertagi antigens and keyhole limpet hemocyanin (KLH) or sheep erythrocytes (SR) for evaluation of antibody production by enzyme linked immunosorbent assay (ELISA) or modified Jerne plaque assay. One semi-purified larval antigen caused both decreased anti-KLH serum antibody levels and fewer anti-SR IgM-secreting B cells. This antigen was shown to depress lymphocyte blastogenesis to Concanavalin A when added to cultured BALB/c splenic lymphocytes.


A finite element analysis of bone plates available for prophylactic internal fixation of the radial osteocutaneous donor site using the sheep tibia model.
The strengthening effect of prophylactic internal fixation (PIF) with a bone plate at the radial osteocutaneous flap donor site has previously been demonstrated using the sheep tibia model of the human radius. This study investigated whether a finite element (FE) model could accurately represent this biomechanical model and whether stress or strain based failure criteria are most appropriate. An FE model of an osteotomised sheep tibia bone was strengthened using 4 types of plates with unilocking or bicortical screw fixation. Torsion and 4-point bending simulations were performed. The maximum von Mises stresses and strain failure criteria were studied. The strengthening effects when applying stress failure criteria [factor 1.76-4.57 bending and 1.33-1.80 torsion] were comparable to the sheep biomechanical model [factor 1.73-2.43 bending and 1.54-2.63 torsion]. The strongest construct was the straight 3.5mm stainless steel unilocking plate. Applying strain criteria the strongest construct was the straight 3.5mm stainless DCP plate with bicortical screw fixation. The FE model was validated by comparison with the sheep tibia model. The complex biomechanics at the bone-screw interface require further investigation. This FE modelling technique may be applied to a model of the human radius and other sites.


Dementia prediction in the general population using clinically accessible variables: a proof-of-concept study using machine learning. The AGES-Reykjavik study.
Early identification of dementia is crucial for prompt intervention for high-risk individuals in the general population. External validation studies on prognostic models for dementia have highlighted the need for updated models. The use of machine learning in dementia prediction is in its infancy and may improve predictive performance. The current study aimed to explore the difference in performance of machine learning algorithms compared to traditional statistical techniques, such as logistic and Cox regression, for prediction of all-cause dementia. Our secondary aim was to assess the feasibility of only using clinically accessible predictors rather than MRI predictors. Data are from 4,793 participants in the population-based AGES-Reykjavik Study without dementia or mild cognitive impairment at baseline (mean age: 76 years, % female: 59%). Cognitive, biometric, and MRI assessments (total: 59 variables) were collected at baseline, with follow-up of incident dementia diagnoses for a maximum of 12 years. Machine learning algorithms included elastic net regression, random forest, support vector machine, and elastic net Cox regression. Traditional statistical methods for comparison were logistic and Cox regression. Model 1 was fit using all variables and model 2 was after feature selection using the Boruta package. A third model explored performance when leaving out neuroimaging markers (clinically accessible model). Ten-fold cross-validation, repeated ten times, was implemented during training. Upsampling was used to account for imbalanced data. Tuning parameters were optimized for recalibration automatically using the caret package in R. 19% of participants developed all-cause dementia. Machine learning algorithms were comparable in performance to logistic regression in all three models. However, a slight added performance was observed in the elastic net Cox regression in the third model (c = 0.78, 95% CI: 0.78-0.78) compared to the traditional Cox regression (c = 0.75, 95% CI: 0.74-0.77). Supervised machine learning only showed added benefit when using survival techniques. Removing MRI markers did not significantly worsen our model's performance. Further, we presented the use of a nomogram using machine learning methods, showing transportability for the use of machine learning models in clinical practice. External validation is needed to assess the use of this model in other populations. Identifying high-risk individuals will amplify prevention efforts and selection for clinical trials.


The redox properties and heme environment of cytochrome c-551.5 from Desulfuromonas acetoxidans.
The environment of the three heme groups in cytochrome c-551.5 from Desulfuromonas acetoxidans was investigated by the technique of solvent perturbation difference spectroscopy. The hemeoctapeptide from cytochrome c plus added imidazole was used as a model compound for the fully exposed chromophore. The average heme exposure in both the ferric and ferrous cytochromes c-551.5 was found to be considerably greater than that previously observed for the monoheme mitochondrial cytochrome c and Prosthecochloris cytochrome c-555. Differences in the average heme exposure for ferric and ferrous cytochromes c-551.5 suggested that a change in oxidation state is accompanied by a change in conformation. A spectrophotometric redox titration of the protein yielded a sigmoidal plot of the potential versus the logarithm of the ratio of oxidized to reduced heme. The resolved plot indicated that two hemes were characterized by a E'o of -177 mV and the third E'o of -102 mV. Each of the resolved steps had an n value of 1 indicating that cytochrome c-551.5 transfers electrons singly.


Abnormal hyperventilation in patients with hepatic cirrhosis: role of enhanced chemosensitivity to carbon dioxide.
Patients with hepatic cirrhosis frequently show idiopathic hyperventilation at rest, despite no concomitant cardiopulmonary disease. The aim of the study was to determine whether altered chemosensitivity either to hypoxia or hypercapnia could underlie inappropriate hyperventilation in cirrhotic patients. We consecutively recruited 30 biopsy proven cirrhotic patients equally distributed in the three Child's classes A, B and C (age 54 ± 8 years, mean ± SD). All patients underwent evaluation of chemosensitivity to hypoxia and to hypercapnia and blood sampling for brain natriuretic peptide, norepinephrine and progesterone, besides full clinical characterization. We also recruited 10 age- and gender-matched healthy controls (age 55 ± 7 years). Overall, 18 patients (60%) showed an increased chemosensitivity to carbon dioxide (CO(2)), while 8 patients (27%) showed enhanced chemosensitivity to hypoxia. Child's class C patients had lower arterial partial pressure of CO(2) (PaCO(2)), higher rest ventilation, increased chemosensitivity to hypercapnia, plasma level of norepinephrine and serum progesterone levels when compared to class A patients and controls (all p < 0.05). Rest ventilation was positively related to pH (R = 0.41, p = 0.023), chemosensitivity to hypercapnia (R = 0.54, p = 0.002), and progesterone (R = 0.53, p = 0.016) and negatively to PaCO(2) (R = 0.61, p < 0.001), but not to hemoglobin level and chemosensitivity to hypoxia. Chemosensitivity to hypercapnia was positively related to PaCO(2) (R = 0.74, p < 0.001), serum progesterone (R = 0.50, p = 0.016), and to plasma norepinephrine (R = 0.57, p = 0.004). Enhanced chemosensitivity to hypercapnia was found in more decompensated cirrhotic patients and was associated with sympathetic overactivity and elevated serum progesterone, likely representing a key mechanism underlying the "unexplained" hyperventilation observed in such patients.


Chemical Profiling of Chlorophytum comosum (Thunb.) Jaques by GC-MS/LC-ESIMS and its Antiproliferative Effects on Human Carcinoma Cell Lines.
Chlorophytum comosum, popularly known as Spider Ivy, is used as a medicinal plant in traditional Chinese medicine, however, its detailed chemical composition and biological activity are yet unexplored. To carry out the phytochemical investigation on different parts of Chlorophytum comosum using GCMS/ LC-ESI-MS and evaluation of its antioxidant, hemolytic and antiproliferative potential on breast cancer (MCF-7), lung cancer (A549, H1299) and normal lung (L-132) cell lines. Chemical constituents from aqueous roots and leaves extracts were identified using LC-ESI-MS/GCMS. The identified compounds were annotated based on the match of mass spectra with the literature using NIST 14 and METLIN databases. Antioxidant activity was studied using DPPH, FRAP and TPC assays. The antiproliferative effects of ethanolic roots and leaves extracts of Chlorophytum comosum were measured by MTT assay on breast cancer (MCF-7), lung cancer (A549 & H1299) and normal lung (L-132) cell lines. The toxicity studies of the extracts were carried out using Hemolysis assay. GC-MS analysis identified 34 metabolites in roots and 17 from leaves, while 17 compounds from roots and 7 from leaves were detected by LC-ESI-MS. Significant antiproliferative effects were observed on the A549 and MCF-7 cancer cell lines with IC50 values ranging from 56.86 μg/ml to 68.68 μg/ml while no marked response was observed against normal cell line L-132. Our study represents the first report on the detailed chemical composition and antiproliferative potential of Chlorophytum comosum against lung and breast cancer cell lines.


Comparison of two titration programmes for adding insulin detemir to oral antidiabetic drugs in patients with poorly controlled type 2 diabetes mellitus.
To explore the effect of active insulin titration versus usual titration on glycaemic control in patients with type 2 diabetes mellitus uncontrolled with oral antidiabetic drugs (OADs). In a 24-week, prospective and randomized study, 172 patients with uncontrolled type 2 diabetes were randomly assigned to either active titration or usual titration. Efficacy and safety outcomes included changes in glycated haemoglobin (HbA1c) and fasting plasma glucose, percentage of individuals achieving HbA1c<53 mmol/mol, and hypoglycaemic events. At Week 24, change in HbA1c was -1.08% ± 1.60% in the active titration group and -0.95% ± 1.34% in the usual titration group (P = 0.569). The percentages of individuals achieving HbA1c<53 mmol/mol were 29.4% and 16.1% in the active and usual titration groups, respectively (P = 0.037). There was no significant difference in the incidence of hypoglycaemia between the two groups. Multivariate logistic regression indicated that, with active titration, baseline HbA1c levels and postprandial glucose excursion were significantly associated with achieving HbA1c<53 mmol/mol. Addition of basal insulin using active titration for 24 weeks provided a higher rate of HbA1c target achievement without significant hypoglycaemia compared to usual titration in individuals with uncontrolled type 2 diabetes.


Midazolam is associated with delay in recovery and agitation after ambulatory general anesthesia for dental treatment in patients with disabilities: a retrospective cohort study.
Some patients with intellectual disabilities (IDs) who undergo total intravenous anesthesia (TIVA) have complications associated with the anesthesia such as prolonged recovery. The purposes of this study were to estimate the frequency of TIVA complications among patients with IDs and to identify factors associated with TIVA complications. This study was designed as a retrospective cohort study. Study samples were selected from the clinical records of patients with IDs who underwent ambulatory general anesthesia in a special dental clinic at the Okayama University Hospital, Okayama, Japan. Predictor variables were patient background, anesthesia-related variables, and dental treatment. Outcome variables were delayed recovery and the complication of agitation. Factors affecting delayed recovery and complications were examined with multivariable analysis. We enrolled 106 cases (81 male and 25 female patients) in this study. The mean age was 23.9 years. Serious complications were not observed in any cases. The amount of intravenous midazolam was an independent determinant of delayed recovery. Oral midazolam contributed to delayed recovery, although it is very useful for induction in patients with a high level of fear. Oral midazolam and a younger age were independent predictors of agitation. Intravenous midazolam may not have an advantage in ambulatory general anesthesia. Oral midazolam contributes to delayed recovery and is an independent predictor of agitation.


Conservative treatment in Jersey finger: a case report.
We report a case of a Type Vb flexor digitorum profundus tendon avulsion in a patient for whom surgery was contraindicated because of a high risk of stroke if anticoagulant treatment was to be stopped. After a 6-week conservative treatment and a 3-week self-physiotherapy programme, the patient achieved a nearly full active range of motion and an excellent functional result. A radiograph demonstrated a good fracture healing. V.


Association of classical and related inflammatory markers with high-sensitivity C-reactive protein in healthy individuals: results from the Stanislas cohort.
Although high-sensitivity C-reactive protein (hs-CRP) has emerged as a cardiovascular marker, questions arise regarding the relative information provided by other inflammatory molecules. Therefore, as a first step, we examined interrelationships between serum hs-CRP concentrations and inflammatory, adhesion and growth factors in healthy adults. Circulating concentrations of hs-CRP, haptoglobin, orosomucoid, interleukin-6 (IL-6), IL-8, IL-18, tumor necrosis factor-alpha (TNF-alpha), TNF-receptor II (TNF-RII), E-, P-, and L-selectins, intercellular adhesion molecule-1 (ICAM-1), monocyte chemoattractant protein-1, endothelial growth factor (EGF), vascular EGF (VEGF), insulin-like growth factor-1 (IGF-1) and IGF-binding protein (IGFBP-3) were measured in 154 men and 161 women of the Stanislas cohort. Leukocyte and platelet counts were also determined. Correlations were significant between hs-CRP concentrations and leukocyte and platelet counts, as well as haptoglobin, orosomucoid, IL-6, and ICAM-1 concentrations (p< or =0.001). Correlation coefficients for ICAM-1 were higher in men than in women (p< or =0.05). When stratifying subjects according to hs-CRP levels, the group with high hs-CRP levels had significantly higher haptoglobin and orosomucoid concentrations than the others, in addition to higher leukocyte counts and IL-6 concentrations in women, and platelet counts and ICAM-1 concentrations in men. Further studies are warranted to explain the association pattern for hs-CRP. Partition of these factors according to their association with hs-CRP concentration opens a new perspective for choice of the best factors in terms of cardiovascular risk in relation to hs-CRP, while non-associated markers could be used to give additional information.


Headache after moderate and severe traumatic brain injury: a longitudinal analysis.
To measure longitudinally headache (HA) after moderate and severe traumatic brain injury (TBI) and to examine potential association with demographic, injury, and psychologic factors. Cohort study. Four Veterans Administration rehabilitation facilities (Minneapolis, Palo Alto, Richmond, Tampa) within the Defense and Veterans Brain Injury Center. Consecutive patients (military or veteran beneficiaries) with moderate or severe TBI (N=109) who during acute rehabilitation consented to data collection and who completed 6- and 12-month follow-up evaluations. Not applicable. HA frequency, location, type, and incapacitation levels measured during prospective neurologic assessments. Nearly 38% (41/109) of patients had acute posttraumatic headache (PTHA) symptoms; most often in a frontal location (20/41), most often of daily frequency (31/41), and showing no relation to injury severity, emotional, or demographic variables. Postacutely, PTHA symptom severity declined within the group. Better individual improvement was associated with less anxiety and depression at 6-month follow-up. Almost all subjects (21/22) with PTHA symptoms that persisted into the 6-month follow-up period reported symptoms again at 12-month follow-up. PTHA severity in this sample of persons with moderate and severe TBI showed a pattern of improvement that leveled off by 6 months posthospitalization.


Correlation between CYP4F2 gene rs2108622 polymorphism and susceptibility to ischemic stroke.
To conduct a meta-analysis for the correlation between cytochrome P450 4F2 (CYP4F2) rs2108622 (V433M) gene polymorphism and ischemic stroke. We retrieved the case-control studies on the correlation between CYP4F2 V433M polymorphism and ischemic stroke included in domestic and international databases before January 2015 and selected the best genetic model, using RevMan 5.2 software for meta-analysis. According to the heterogeneity test results of selected literature, the effect model of consolidated data was selected, and the combined OR and 95% CI were calculated. A total of six documents were included. Recessive model (VM + MM vs. VV) was selected as the best genetic model. The combined results showed that: compared with wild-type VV, there are significant association between ischemic stroke and CYP4F2 polymorphism (OR merge = 1.37, 95% CI: 1.21~1.54, P < 0.001). CYP4F2 V433M may be the susceptibility gene for ischemic stroke.


[Injury of myocardial mitochondria in neonatal swines with hypoxic-ischemic brain damage].
To clarify the effect of hypoxia and ischemia on the concentration of Ca2+ in myocardial mitochondria, and on the activity of respiratory chain complex IV--cytochrome oxidase (CCO) in neonatal swines, and to find out the change of myocardial mitochondria during hypoxic-ischemic brain damage (HIBD). The study was performed on neonatal swines. For establishment of HIBD model, the left carotid was ligated and the swine was placed under hypoxia for two hours. Then the concentration of Ca2+ and the activity of respiratory chain complex IV in myocardial cell mitochondria were measured at 0 hour and 24, 48, 72 hours after reoxygenation. The measured values from these 0-, 24-, 48-, 72-hour-HIBD-groups (n = 10 per group) and from the normal control group (n = 10) were statistically analyzed by means of ANOVA, Newman-Keuls test and linear correlation. After the swines were placed under hypoxia-ischemia for two hours, 1. the concentration of Ca2+ in myocardial mitochondria increased, till 24 hours after re-oxygenation, and then it descended at 48 hours and 72 hours after reoxygenation; 2. the activity of myocardial mitochondrial respiratory chain complex IV descended, though activity recovery was seen at 48 and 72 hours, it did not come back to normal level at 72 hours after reoxygenation; 3. the concentration of Ca2+ in myocardial mitochondria was negatively correlated with the activity of respiratory chain complex IV. The results showed that in the neonatal swines with HIBD, the mitochondrial respiratory chain in myocardial cell was damaged by hypoxia-ischemia, yet it could be rehabilitated after reoxygenation. Calcium overload in myocardial mitochondria after hypoxia-ischemia might be one of the causes of the descending activity of respiratory chain complex IV.


μ opioid receptor agonist-selective regulation of interleukin-4 in T lymphocytes.
Opioids are irreplaceable for the treatment of severe pain. However, opioid-induced immunomodulation affects therapies. Here we report that treatment of human T lymphocytes with the opioids fentanyl, methadone, loperamide and beta-endorphin resulted in a strong induction of the cytokine interleukin-4. In contrast, morphine and buprenorphine induced markedly and significantly lower levels of interleukin-4 mRNA and protein. These findings suggest agonist-biased μ opioid receptor signaling in T cells. In the future, better knowledge about agonist-specific immunomodulatory effects of opioids offers the possibility to select drugs for a therapy with more favorable and/or less detrimental side effects in immune cells.


End points in dermatologic clinical trials: A review for clinicians.
Clinical trials are critical for the development of new therapies in dermatology, and their results help determine US Food and Drug Administration (FDA) approval and guide care. Of special relevance is the clinical trial efficacy end point, the metric from which statistically significant outcome is derived. Clinicians' understanding of a clinical trial's end point is necessary for critical analysis of the trial results and for applying those results to daily practice. This review provides practical knowledge and critical evaluation of end points used in treatment approvals by the FDA. The end points for actinic keratosis, acne vulgaris, atopic dermatitis, onychomycosis, and cutaneous ulcer serve as examples.


ATTR Amyloidosis: Current and Emerging Management Strategies: JACC: CardioOncology State-of-the-Art Review.
Transthyretin cardiac amyloidosis (ATTR-CA) is increasingly diagnosed owing to the emergence of noninvasive imaging and improved awareness. Clinical penetrance of pathogenic alleles is not complete and therefore there is a large cohort of asymptomatic transthyretin variant carriers. Screening strategies, monitoring, and treatment of subclinical ATTR-CA requires further study. Perhaps the most important translational triumph has been the development of effective therapies that have emerged from a biological understanding of ATTR-CA pathophysiology. These include recently proven strategies of transthyretin protein stabilization and silencing of transthyretin production. Data on neurohormonal blockade in ATTR-CA are limited, with the primary focus of medical therapy on judicious fluid management. Atrial fibrillation is common and requires anticoagulation owing to the propensity for thrombus formation. Although conduction disease and ventricular arrhythmias frequently occur, little is known regarding optimal management. Finally, aortic stenosis and ATTR-CA frequently coexist, and transcatheter valve replacement is the preferred treatment approach.


Interobserver agreement and efficacy of consensus reading in Kwak-, EU-, and ACR-thyroid imaging recording and data systems and ATA guidelines for the ultrasound risk stratification of thyroid nodules.
To investigate the interobserver agreement (IA) and the impact of consensus reading using four risk stratification systems for thyroid nodules (TN). Four experienced specialists independently rated US images of 80 TN according to the Kwak-TIRADS, EU-TIRADS, ACR TI-RADS, and ATA Guidelines. The cases were randomly extracted from a prospectively acquired database (n > 1500 TN). The observers were blinded to clinical data. This study was divided into two sessions (S1 and S2) with 40 image sets each. After every session, a consensus reading was carried out (C1, C2). Subsequently, the effect of C1 was tested in S2 with 40 new cases followed by C2. Fleiss' kappa (κ) was calculated for S1 and S2 to estimate the IA and learning curves. The results of C1 and C2 were used as reference for diagnostic accuracy calculations. IA significantly increased (p < 0.01) after C1 with κ values of 0.375 (0.615), 0.411 (0.596), 0.321 (0.569), and 0.410 (0.583) for the Kwak-TIRADS, EU-TIRADS, ACR TI-RADS, and ATA Guidelines in S1 (S2), respectively. ROC analysis (C1 + C2) revealed similar areas under the curve (AUC) for the Kwak-TIRADS, EU-TIRADS, ACR TI-RADS, and ATA Guidelines (0.635, 0.675, 0.694, and 0.654, respectively, n.s.). AUC did not increase from C1 (0.677 ± 0.010) to C2 (0.632 ± 0.052, n.s.). ATA Guidelines were not applicable in five cases. IA and diagnostic accuracy were very similar for the four investigated risk stratification systems. Consensus reading sessions significantly improved the IA but did not affect the diagnostic accuracy.


The use of RA 233 in the preparation of platelets, with the blood cell separator.
The intravenous administration of the inhibitor of platelet adhesiveness RA 233 to blood donors before and during cell separation did not increase the yield of platelets in the final concentrate.


Atrial fibrillation prediction by combining ECG markers and CMR radiomics.
Atrial fibrillation (AF) is the most common cardiac arrhythmia. It is associated with a higher risk of important adverse health outcomes such as stroke and death. AF is linked to distinct electro-anatomic alterations. The main tool for AF diagnosis is the Electrocardiogram (ECG). However, an ECG recorded at a single time point may not detect individuals with paroxysmal AF. In this study, we developed machine learning models for discrimination of prevalent AF using a combination of image-derived radiomics phenotypes and ECG features. Thus, we characterize the phenotypes of prevalent AF in terms of ECG and imaging alterations. Moreover, we explore sex-differential remodelling by building sex-specific models. Our integrative model including radiomics and ECG together resulted in a better performance than ECG alone, particularly in women. ECG had a lower performance in women than men (AUC: 0.77 vs 0.88, p < 0.05) but adding radiomics features, the accuracy of the model was able to improve significantly. The sensitivity also increased considerably in women by adding the radiomics (0.68 vs 0.79, p < 0.05) having a higher detection of AF events. Our findings provide novel insights into AF-related electro-anatomic remodelling and its variations by sex. The integrative radiomics-ECG model also presents a potential novel approach for earlier detection of AF.


Evaluation of Immunophenotypic and Molecular Biomarkers for Sézary Syndrome Using Standard Operating Procedures: A Multicenter Study of 59 Patients.
Differentiation between Sézary syndrome and erythrodermic inflammatory dermatoses can be challenging, and a number of studies have attempted to identify characteristic immunophenotypic changes and molecular biomarkers in Sézary cells that could be useful as additional diagnostic criteria. In this European multicenter study, the sensitivity and specificity of these immunophenotypic and recently proposed but unconfirmed molecular biomarkers in Sézary syndrome were investigated. Peripheral blood CD4(+) T cells from 59 patients with Sézary syndrome and 19 patients with erythrodermic inflammatory dermatoses were analyzed for cell surface proteins by flow cytometry and for copy number alterations and differential gene expression using custom-made quantitative PCR plates. Experiments were performed in duplicate in two independent centers using standard operating procedures with almost identical results. Sézary cells showed MYC gain (40%) and MNT loss (66%); up-regulation of DNM3 (75%), TWIST1 (69%), EPHA4 (66%), and PLS3 (66%); and down-regulation of STAT4 (91%). Loss of CD26 (≥80% CD4(+) T cells) and/or CD7 (≥40% CD4(+) T cells) and combination of altered expression of STAT4, TWIST1, and DNM3 or PLS3 could distinguish, respectively, 83% and 98% of patients with Sézary syndrome from patients with erythrodermic inflammatory dermatoses with 100% specificity. These additional diagnostic panels will be useful adjuncts in the differential diagnosis of Sézary syndrome versus erythrodermic inflammatory dermatoses.


Novel metabotropic glutamate receptor negatively coupled to adenylyl cyclase in cultured rat cerebellar astrocytes.
Several excitatory amino acid ligands were found potently to inhibit forskolin-stimulated cAMP accumulation in rat cultured cerebellar astrocytes: L-cysteine sulfinic acid (L-CSA) = L-aspartate > L-glutamate >/= the glutamate uptake inhibitor, L-PDC. This property did not reflect activation of conventional glutamate receptors, since the selective ionotropic glutamate receptor agonists NMDA, AMPA, and kainate, as well as several mGlu receptor agonists [(1S,3R)-ACPD, (S)-DHPG, DCG-IV, L-AP4, L-quisqualate, and L-CCG-I], were without activity. In addition, the mGlu receptor antagonists, L-AP3, (S)-4CPG, Eglu, LY341495, (RS)-CPPG, and (S)-MCPG failed to reverse 30 microM glutamate-mediated inhibitory responses. L-PDC-mediated inhibition was abolished by the addition of the enzyme glutamate-pyruvate transaminase. This finding suggests that the effect of L-PDC is indirect and that it is mediated through endogenously released L-glutamate. Interestingly, L-glutamate-mediated inhibitory responses were resistant to pertussis toxin, suggesting that G(i)/G(o) type G proteins were not involved. However, inhibition of protein kinase C (PKC, either via the selective PKC inhibitor GF109203X or chronic PMA treatment) augmented glutamate-mediated inhibitory responses. Although mGlu3 receptors (which are negatively coupled to adenylyl cyclase) are expressed in astrocyte populations, in our study Western blot analysis indicated that this receptor type was not expressed in cerebellar astrocytes. We therefore suggest that cerebellar astrocytes express a novel mGlu receptor, which is negatively coupled to adenylyl cyclase, and possesses an atypical pharmacological profile.


Spiroheterocyclic Photocatalyst for Reducing QHIn-Persistent Pollutants, Dyes, and Transition-Metal Ions Cocatalyzed with Electrolytes.
The 7-nitro-2'-phenyl-5',6',7',7a'-tetrahydrospiro[indeno[1,2-b]quinoxaline-11,3'-pyrrolizine]-1',1'(2'H)-dicarbonitrile (SIQPI), 2'-(4-cyanophenyl)-7-nitro-5',6',7',7a'-tetrahydrospiro[indeno[1,2-b]quinoxaline-11,3'-pyrrolizine]-1',1'(2'H) dicarbonitrile (SIQPII), and 2'-(4-methoxyphenyl)-7-nitro-5',6',7',7a'-tetrahydrospiro[indeno[1,2-b]quinoxaline-11,3'-pyrrolizine]-1',1'(2'H)-dicarbonitrile (SIQPIII) were used to photocatalyze quinonoid phenolphthalein (QHIn) in aq-ACN-EtOH (mixed solvent) with NaCl and KCl electrolytes. SIQPI, II, and III spiroindenoquinoxaline pyrrolidines (SIQPs) as spiroheterocyclic photocatalysts alone could not reduce QHIn, but with the addition of electrolytes they are reduced via π cationic interactions (PCI). SIQPI, II, and III with NaCl reduced QHIn in 120, 28, and 50 min, unlike in 138, 58, and 63 min with KCl in mixed solvent. SIQPI, II, and III alone have reduced methylene blue (MB) in 120, 45, and 70 min, unlike in 110, 27, and 55 min with graphene oxide (GO), whereas with NaCl and KCl hey are reduced in 82, 36, and 44 min and 89, 43, and 50 min, respectively. SIQPs with GO had reduced MB in less time than the SIQPs alone, and SIQPs with NaCl had reduced QHIn in a shorter time than KCl. The electrolytes have cocatalyzed a reduction of dyes under sunlight (SL). The electrolytes have reduced a quinonoid structure (QS) and dyes by generating negative and positive (e - and h +) holes in a shorter time. SIQPII and magnetic nanoparticles (MNPs) of 58 nm with NaCl photocatalyzed the QHIn in 2880 min. The SIQPs also reduced methyl orange (MO) and brilliant blue R (BBR) at variable temperature (T) and pH range, whereas SIQPs have developed a molecular organic framework (MOF) with transition-metal salts (NiCl2, CrO3, KMnO4, CuSO4, and MnCl2) on photocatalysis.


Transgender and Gender Diverse Patients in Intensive Mood Disorder Treatment: A Comparative Examination of Clinical Presentation and Treatment Outcomes.
Individuals who are transgender and gender diverse (TGD) are more likely to suffer from and to seek mental health services for mood disorders. Some literature suggests that TGD individuals, because of pervasive and systemic minority stress, may have more complex clinical presentations (i.e., psychiatric conditions and severity of symptoms) and may benefit from empirically based treatments to a lesser degree than their cisgender peers. However, research has yet to examine individuals who are TGD receiving treatment in specialized, intensive mood disorder treatment despite the propensity for them to be diagnosed with and treated for mood disorders. Using a sample of 1,326 adult patients in intensive mood disorder treatment (3.8% TGD), the clinical presentation and treatment outcomes were compared between patients who are TGD and cisgender. Contrary to previous research, TGD patients were largely similar if not healthier than their cisgender counterparts, including similar depression severity, quality of life, emotion dysregulation, and behavioral activation, and less severe rumination at admission. Despite similar to better reported mental health symptoms, TGD patients were diagnosed with more psychiatric conditions overall, including greater prevalence of social anxiety and neurodevelopmental diagnoses. Those who are TGD did not experience attenuated treatment response as predicted. Findings suggest that patients in intensive mood disorder treatment who are TGD may be more resilient than previously assumed, or supports may have increased to buffer effects of stigma on mental health, and emphasize the need to exercise discretion and sensitivity in diagnostic practices to prevent over-diagnosis and pathologizing of TGD individuals.


Non-tryptophan fluorescence and high molecular weight protein formation in lens crystallins of rats with chronic galactosemia: prevention by the aldose reductase inhibitor sorbinil.
Crystallin-bound non-tryptophan fluorescence and protein cross-linking were studied in chronic galactosemia. Fluorescence (excitation/emission--370/440 nm) was significantly higher in galactosemic rats compared to controls (P less than 0.001). Accumulation of fluorescence was significantly reduced both in water soluble (P less than 0.001) and insoluble (P less than 0.005) lens fractions in galactosemic rats receiving the aldose reductase inhibitor sorbinil. High performance liquid chromatography of water soluble lens crystallins showed an increase in the content of high molecular weight proteins and the fluorescence associated with it. Sorbinil partly prevented the formation of such fluorescent high molecular weight proteins. Under reducing conditions, sodium dodecyl sulfate polyacrylamide gel electrophoresis of water soluble proteins revealed a distinct high molecular weight protein of 60 kDa in galactosemia. Sorbinil treatment completely abolished the formation of this protein. Western blotting using rabbit antiserum to bovine alpha-, beta- and gamma-crystallins revealed the presence of gamma, but not alpha- and beta-crystallins in the 60-kDa protein. Formation of this protein may result from trimerization of gamma-crystallins or from an association of gamma-crystallin with a non-crystallin cytosolic protein or mixed protein cross-linking of gamma-crystallins with membrane protein(s). The present study shows that polyol pathway in cataractogenesis also extends to protein cross-linking and formation of fluorescent compounds, the nature of which remains to be elucidated.


Unilateral triplication of the collecting system in a horseshoe kidney.
We report on complete unilateral triplication of the collecting system and ureters in a patient with a horseshoe kidney. One of the ureteral segments was obstructed and the patient presented with urosepsis. To our knowledge, such a malformation has not been described previously.


Three-year surveillance of culicine mosquitoes (Diptera: Culicidae) for flavivirus infections in Incheon Metropolitan City and Hwaseong-si of Gyeonggi-do Province, Republic of Korea.
Japanese encephalitis virus (JEV) is a single stranded positive sense RNA virus of the genus Flavivirus that belongs to family Flaviviridae and emerged as one of the most pivotal form of viral encephalitis. The virus is transmitted to humans by mosquito vector and is an etiological agent of acute zoonotic infection. In this study, we investigated distribution and density over 3-year period in central regions of Korean peninsula. We selected two cities as mosquito-collecting locations and subdivided them into five collection sites; downtown Incheon Metropolitan City as a typical urban area, and the Hwaseong-si area as a rural area. A total of 35,445 female culicine mosquitoes were collected using black light traps or BG Sentinel™ traps from March to November 2016-2018. Aedes (Ae.) vexans nipponii was the most frequently collected specimens (48.91%), followed by Culex (Cx.) pipiens (32.05%), Ochlerotatus (Och.) dorsalis (13.58%), Och. koreicus (1.68%), and Cx. tritaeniorhynchus (1.49%). In the urban area, Cx. pipiens was the predominant species (92.21%) and the other species accounted for <5% of the total mosquitoes collected. However, in the rural area, Ae. vexans nipponii had the highest population (61.90%), followed by Och. dorsalis (17.10%), Cx. tritaeniorhynchus (1.84%) and Och, koreicus (1.78%). Culicine mosquitoes were identified at the species level, placed in pools of up to 30 mosquitoes each, and screened for flavivirus RNA using the SYBR Green-based RT-PCR. Three of the assayed 1092 pools were positive for Chaoyang virus from Ae. vexans nipponii and Japanese encephalitis virus from Cx. pipiens. The maximum likelihood estimations (the estimated number of virus-positive mosquitoes/1000 mosquitoes) for Ae. vexans nipponii positive for Chaoyang virus and Cx. pipiens for Japanese encephalitis virus were 3.095 and 0.20, respectively. The results of our study demonstrate that although mosquito-borne diseases were not detected in the potential vectors, enhanced monitoring and long-term surveillance of these vector viruses are of great public health importance.


Ventilatory responses to hypercapnia and hypoxia following chronic hypercapnia in the rat.
This study investigated the effects of an 18 week exposure to 10% CO(2) in air on minute ventilation (V(E)), breathing pattern and the chemoresponiveness of rats to hypoxic and hyperoxic stimuli. We found that V(E) remained elevated over the 18 weeks. Nonetheless, the breathing pattern changed significantly. Tidal volume increased and the durations of inspiration and the total cycle decreased. After the sustained hypercapnia the mean Pa(CO(2)) was 72.0+/-5.1 (S. D.) mmHg. Every 6 weeks the chemoresponiveness of the CO(2)-exposed rats was tested by an acute exposure sequentially to room air, then a 6% O(2), 10% CO(2) and 84% N(2) gas mixture, and finally a 90% O(2) in 10% CO(2) mixture. On either room air or the hyperoxic-hypercapnic mixture V(E) fell to its pre-hypercapnic level. On the hypoxic-hypercapnic mixture V(E) increased significantly. These results demonstrate that the initial stimulating effect of 10% CO(2) on V(E) persisted for the entire 18 weeks without altering hypoxic or hyperoxic ventilatory responses.


Use of fungal and bacterial protease preparations to enhance extraction of lipid from fish roe: Effect on lipidomic profile of extracted oil.
The present study investigated the hydrolysis of protein in hoki roe homogenate using a HT (bacterial), a FP-II (fungal) protease preparations and Alcalase (bacterial) to enhance lipid yield extraction. The degree of hydrolysis was determined at various pH, temperature and time using casein and hoki roe. Total lipid extraction and lipidomic analysis was carried out following proteolysis of hoki roe homogenate. The degree of hydrolysis and SDS-PAGE revealed that the hydrolytic capability of Alcalase was better than HT and FPII. The total extracted lipid yield was better following hydrolysis with Alcalase (19.29 %), compared to HT (18.29 %) and FPII (18.33 %). However, the total phospholipid (PL) and n-3 fatty acid yields were better from HT hydrolysed hoki roe homogenate (PL = 30.7 μmol/g; n-3 = 10.5 %), compared to Alcalase (PL = 22 μmol/g; n-3 = 5.95 %). Overall, this study indicates that HT protease preparation hydrolysis of fish roe homogenate can both enhance lipid extraction and retain lipid quality.


An uncommonly serious case of an uncommon sport injury.
A 55 year old man sustained a severe ocular injury when hit by a cricket ball even though he was wearing a helmet. A suprachoroidal haemorrhage was drained and dense intravitreal blood was removed. An inferior buckle was applied with the use of intraocular gas. A macular haemorrhage resolved slowly. Despite several surgical procedures over 1.5 years, the final visual acuity of the patient was only 6/60 because of a dense macular scar. Helmets worn as protection when playing cricket need to be designed better and be of better material. Eye protection should be worn at all levels of play.