Sex and β-Endorphin Influence the Effects of Ethanol on Limbic Gabra2 Expression in a Mouse Binge Drinking Model.
Binge drinking is a widespread problem linked to increased risk for alcohol-related complications, including development of alcohol use disorders. In the last decade, binge drinking has increased significantly, specifically in women. Clinically, sexually dimorphic effects of alcohol are well-characterized, however, the underlying mechanisms for these dimorphisms in the physiological and behavioral effects of alcohol are poorly understood. Among its many effects, alcohol consumption reduces anxiety via the inhibitory neurotransmitter GABA, most likely acting upon receptors containing the α-2 subunit (Gabra2). Previous research from our laboratory indicates that female mice lacking the endogenous opioid peptide β-endorphin (βE) have an overactive stress axis and enhanced anxiety-like phenotype, coupled with increased binge-like alcohol consumption. Because βE works via GABA signaling to reduce anxiety, we sought to determine whether sexually dimorphic binge drinking behavior in βE deficient mice is coupled with differences in CNS Gabra2 expression. To test this hypothesis, we used βE knock-out mice in a "drinking in the dark" model where adult male and female C57BL/6J controls (βE +/+) and βE deficient (βE -/-; B6.129S2-Pomctm1Low/J) mice were provided with one bottle of 20% ethanol (EtOH) and one of water (EtOH drinkers) or two bottles of water (water drinkers) 3 h into the dark cycle for four consecutive days. Following a binge test on day 4, limbic tissue was collected and frozen for subsequent qRT-PCR analysis of Gabra2 mRNA expression. Water-drinking βE +/+ females expressed more Gabra2 in central nucleus of the amygdala and the bed nucleus of the stria terminalis than males, but this sex difference was absent in the βE -/- mice. Genotype alone had no effect on alcohol consumption or drug-induced increase in Gabra2 expression. In contrast, βE expression had bi-directional effects in females: in wildtypes, Gabra2 mRNA was reduced by binge EtOH consumption, while EtOH increased expression in βE -/- females to levels commensurate with drug-naïve βE +/+ females. These results support the contention that βE plays a role in sexually dimorphic binge-like EtOH consumption, perhaps through differential expression of GABAA α2 subunits in limbic structures known to play key roles in the regulation of stress and anxiety.


The association of statin subgroups with lower urinary tract symptoms following a prostate biopsy.
This was a secondary analysis aiming to assess whether hydrophilic or hydrophobic statins have a differential effect on urinary retention (UR) and lower urinary tract symptoms (LUTS) in men following a prostate biopsy (PBx), who were at risk for prostate cancer development. This was a population-based cohort study with data incorporated from the Institute for Clinical and Evaluative Sciences database to identify all Ontarian men aged 66 and above with a history of a single negative PBx between 1994 and 2016, with no drug prescription history of any of several putative chemo-preventative medications (statins, proton pump inhibitors, five-alpha-reductase inhibitors, and alpha-blockers). Multivariable Cox regression models with time-dependent covariates were used to assess the association of hydrophilic and hydrophobic statins with UR and LUTS within 30 days of a PBx. All models were adjusted for other known putative chemopreventive medications, age, rurality, pharmacologically treated diabetes, comorbidity score, and study inclusion year. Overall, 21 512 men were included, with a median followup time of 9.4 years (interquartile range [IQR] 5.4-13.4 years). Hydrophobic and hydrophilic statins were initiated by 30.7% and 19.6% of men, respectively, after the first negative PBx. UR and LUTS were experienced by 2.2% and 10% of men, respectively. Cox models demonstrated hydrophilic statins were associated with a lower risk of UR (hazard ratio [HR] 0.56, 95% confidence interval [CI] 0.38-0.83, p=0.0038) and LUTS (HR 0.86, 95% CI 0.76-0.98, p=0.022), while no such association was shown for hydrophobic statins. Initiation of hydrophilic statins in men older than 66 appears to be inversely associated with the risk of UR and LUTS within 30 days of a PBx.


On the molecular structure of human neuroserpin polymers.
The polymerization of serpins is at the root of a large class of diseases; the molecular structure of serpin polymers has been recently debated. In this work, we study the polymerization kinetics of human neuroserpin by Fourier Transform Infra Red spectroscopy and by time-lapse Size Exclusion Chromatography. First, we show that two distinct neuroserpin polymers, formed at 45 and 85°C, display the same isosbestic points in the Amide I' band, and therefore share common secondary structure features. We also find a concentration independent polymerization rate at 45°C suggesting that the polymerization rate-limiting step is the formation of an activated monomeric species. The polymer structures are consistent with a model that predicts the bare insertion of portions of the reactive center loop into the A β-sheet of neighboring serpin molecule, although with different extents at 45 and 85°C.


Subtle changes in hydrogen bond orientation result in glassification of carbon capture solvents.
Water-lean CO2 capture solvents show promise for more efficient and cost-effective CO2 capture, although their long-term behavior in operation has yet to be well studied. New observations of extended structure solvent behavior show that some solvent formulations transform into a glass-like phase upon aging at operating temperatures after contact with CO2. The glassification of a solvent would be detrimental to a carbon-capture process due to plugging of infrastructure, introducing a critical need to decipher the underlying principles of this phenomenon to prevent it from happening. We present the first integrated theoretical and experimental study to characterize the nano-structure of metastable and glassy states of an archetypal single-component alkanolguanidine carbon-capture solvent and assess how minute changes in atomic-level interactions convert the solvent between metastable and glass-like states. Small-angle neutron scattering and neutron diffraction coupled with small- and wide-angle X-ray scattering analysis demonstrate that minute structural changes in solution precipitae reversible aggregation of zwitterionic alkylcarbonate clusters in solution. Our findings indicate that our test system, an alkanolguanidine, exhibits a first-order phase transition, similar to a glass transition, at approximately 40 °C-close to the operating absorption temperature for post-combustion CO2 capture processes. We anticipate that these phenomena are not specific to this system, but are present in other classes of colvents as well. We discuss how molecular-level interactions can have vast implications for solvent-based carbon-capture technologies, concluding that fortunately in this case, glassification of water-lean solvents can be avoided as long as the solvent is run above its glass transition temperature.


A patient with an inborn error of vitamin B12 metabolism (cblF) detected by newborn screening.
A neonate, who was found to have an elevated C3/C2 ratio and minimally elevated propionylcarnitine on newborn screening, was subsequently identified as having the rare cblF inborn error of vitamin B12 (cobalamin) metabolism. This disorder is characterized by the retention of unmetabolized cobalamin in lysosomes such that it is not readily available for cellular metabolism. Although cultured fibroblasts from the patient did not show the expected functional abnormalities of the cobalamin-dependent enzymes, methylmalonyl-CoA mutase and methionine synthase, they did show reduced synthesis of the active cobalamin cofactors adenosylcobalamin and methylcobalamin. Mutation analysis of LMBRD1 established that the patient had the cblF disorder. Treatment was initiated promptly, and the patient showed a robust response to regular injections of cyanocobalamin, and she was later switched to hydroxocobalamin. Currently, at 3 years of age, the child is clinically well, with appropriate development. Adjusted newborn screening cutoffs in Ontario allowed detection of a deficiency that might not have otherwise been identified, allowing early treatment and perhaps preventing the adverse sequelae seen in some untreated patients.


Mechanism underlying polyvalent IgG-induced regulatory T cell activation and its clinical application: Anti-idiotypic regulatory T cell theory for immune tolerance.
The regulatory T (Treg) cells constitute a functionally defined subpopulation of T cells that modulate the immune system and maintain immune tolerance through suppression of the development of autoimmune responses to self-antigens and allergic reactions to external antigens. Reduction in the number or function of Treg cells has been suggested as a key immune abnormality underlying the development of autoimmune and allergic diseases. In vitro studies have demonstrated that purified polyvalent immunoglobulin G (IgG) from multiple healthy blood donors can exert immunomodulatory effects on Treg cells. Incubation of polyvalent human IgG with purified CD4+CD25high T cells increased the intracellular expression of interleukin (IL)-10. Intravenous administration of polyvalent human IgG induced significant expansions of CD4+ Foxp3+ Treg cells and clinical improvements in patients with autoimmune diseases. In human clinical trials, intramuscular administration of autologous total IgG significantly increased the percentage of IL-10-producing CD4+ Treg cells in the peripheral blood of healthy subjects and provided significant clinical improvements in patients with atopic dermatitis. These results suggest a clinical usefulness of polyvalent IgG-induced activation of Treg cells in human subjects. This review proposes a new hypothesis for immune tolerance mechanism by integrating the pre-existing "idiotypic network theory" and "Treg cell theory" into an "anti-idiotypic Treg cell theory." Based on this hypothesis, an "active anti-idiotypic therapy" for allergic and autoimmune diseases using autologous polyvalent IgG (as immunizing antigens) is suggested as follows: (1) Intramuscular or subcutaneous administration of autologous polyvalent IgG produces numerous immunogenic peptides derived from idiotypes of autologous IgG through processing of dendritic cells, and these peptides activate anti-idiotypic Treg cells in the same subject. (2) Activated anti-idiotypic Treg cells secrete IL-10 and suppress Th2 cell response to allergens and autoimmune T cell response to self-antigens. (3) These events can induce a long-term clinical improvements in patients with allergic and autoimmune diseases. Further studies are needed to evaluate the detailed molecular mechanism underlying polyvalent IgG-induced Treg cell activation and the clinical usefulness of this immunomodulatory therapy for autoimmune and allergic diseases.


Translational Antiphase Boundaries in NaNbO3 Antiferroelectrics.
Planar defects are known to be of importance in affecting the functional properties of materials. Translational antiphase boundaries (APBs) in particular have attracted considerable attention in perovskite oxides, but little is known in lead-free antiferroelectric oxides that are promising candidates for energy storage applications. Here, we present a study of translational APBs in prototypical antiferroelectric NaNbO3 using aberration-corrected (scanning) transmission electron microscopy (TEM) techniques at different length scales. The translational APBs in NaNbO3 are characterized by a 2-fold-modulated structure, which is antipolar in nature and exhibits a high density, different from the polar nature and lower density in PbZrO3. The high stability of translational APBs against external electric fields and elevated temperatures was revealed using ex situ and in situ TEM experiments and is expected to be associated with their antipolar nature. Density functional theory calculations demonstrate that translational APBs possess only slightly higher free energy than the antiferroelectric and ferroelectric phase energies with differences of 29 and 33 meV/f.u., respectively, justifying their coexistence down to the nanoscale at room temperature. These results provide a detailed atomistic elucidation of translational APBs in NaNbO3 with antipolar character and stability against external stimuli, establishing the basis of defect engineering of antiferroelectrics for energy storage devices.


Evaluation of nine automated high-sensitivity C-reactive protein methods: implications for clinical and epidemiological applications. Part 2.
C-Reactive protein (CRP) can provide prognostic information about risk of future coronary events in apparently healthy subjects. This application requires higher sensitivity assays than have traditionally been available in the clinical laboratory. Nine high-sensitivity CRP (hs-CRP) methods from Dade Behring, Daiichi, Denka Seiken, Diagnostic Products Corporation, Iatron, Kamiya, Olympus, Roche, and Wako were evaluated for limit of detection, linearity, precision, prozone effect, and comparability with samples from 388 apparently healthy individuals. All methods had limits of detection that were lower than the manufacturers' claimed limit of quantification except for the Kamiya, Roche, and Wako methods. All methods were linear at 0.3-10 mg/L. The Diagnostic Products Corporation, Kamiya, Olympus, and Wako methods had imprecision (CVs) >10% at 0.15 mg/L. The Iatron, Olympus, and Wako methods demonstrated prozone effects at hs-CRP concentrations of 12, 206, and 117 mg/L, respectively. hs-CRP concentrations demarcating each quartile in a healthy population were method-dependent. Ninety-two to 95% of subjects were classified into the same quartile of hs-CRP established by the Dade Behring method by the Denka Seiken, Diagnostic Products Corporation, Iatron, and Wako methods. In contrast, 68-77% of subjects were classified into the same quartile by the Daiichi, Kamiya, Olympus, and Roche methods. No subject varied by more than one quartile by any method. Four of the nine examined hs-CRP methods classified apparently healthy subjects into quartiles of hs-CRP similar to the classifications assigned by the comparison method. Additional standardization efforts are required because an individual patient's results will be interpreted using population-based cutpoints.


Rapid ventricular pacing during transcatheter valve procedures using an internal device and programmer: A demonstration of feasibility.
To develop a protocol for using a pre-existing, permanent pacemaker or defibrillator device for rapid ventricular pacing during transcatheter valve procedures and demonstrate feasibility. Placement of a passive fixation, temporary pacemaker wire is considered routine during most transcatheter valve procedures to facilitate controlled or rapid ventricular pacing at the time of balloon expansion or valve deployment. Many patients presenting for such procedures have a pre-existing, permanent pacemaker or defibrillator device which could be used for the same function, obviating the need for temporary pacemaker wire placement. We developed a strategy for rapid pacing from the pre-existing device using a programmer during transcatheter valve procedures in consecutive patients over a 3-month period. Complications and clinical outcomes were recorded. There were 135 transcatheter valve procedures performed during the study. Of these, 28 (20.7%) had pre-existing devices (17 transcatheter aortic valve replacement, 3 aortic valve-in-valve, 2 mitral valve-in-valve, and 6 balloon aortic valvuloplasty). All patients underwent rapid ventricular pacing using a commercially available device programmer. There were no adverse events related to device pacing and no patients required placement of a temporary pacemaker wire during the procedure. At 30-days follow-up, there were no deaths, one major vascular complication related to arterial access, and one patient with renal failure requiring dialysis. Pacing from a commercially available device programmer is safe, feasible, and may reduce both procedural cost and complications such as cardiac tamponade by avoiding placement of a temporary pacemaker lead during transcatheter valve procedures.


Brain insulin receptor causes activity-dependent current suppression in the olfactory bulb through multiple phosphorylation of Kv1.3.
Insulin and insulin receptor (IR) kinase are found in abundance in discrete brain regions yet insulin signaling in the CNS is not understood. Because it is known that the highest brain insulin-binding affinities, insulin-receptor density, and IR kinase activity are localized to the olfactory bulb, we sought to explore the downstream substrates for IR kinase in this region of the brain to better elucidate the function of insulin signaling in the CNS. First, we demonstrate that IR is postnatally and developmentally expressed in specific lamina of the highly plastic olfactory bulb (OB). ELISA testing confirms that insulin is present in the developing and adult OB. Plasma insulin levels are elevated above that found in the OB, which perhaps suggests a differential insulin pool. Olfactory bulb insulin levels appear not to be static, however, but are elevated as much as 15-fold after a 72-h fasting period. Bath application of insulin to cultured OB neurons acutely induces outward current suppression as studied by the use of traditional whole-cell and single-channel patch-clamp recording techniques. Modulation of OB neurons is restricted to current magnitude; IR kinase activation does not modulate current kinetics of inactivation or deactivation. Transient transfection of human embryonic kidney cells with cloned Kv1.3 ion channel, which carries a large proportion of the outward current in these neurons, revealed that current suppression was the result of multiple tyrosine phosphorylation of Kv1.3 channel. Y to F single-point mutations in the channel or deletion of the kinase domain in IR blocks insulin-induced modulation and phosphorylation of Kv1.3. Neuromodulation of Kv1.3 current in OB neurons is activity dependent and is eliminated after 20 days of odor/sensory deprivation induced by unilateral naris occlusion at postnatal day 1. IR kinase but not Kv1.3 expression is downregulated in the OB ipsilateral to the occlusion, as demonstrated in cryosections of right (control) and left (sensory-deprived) OB immunolabeled with antibodies directed against these proteins, respectively. Collectively, these data support the hypothesis that the hormone insulin acts as a multiply functioning molecule in the brain: IR signaling in the CNS could act as a traditional growth factor during development, be altered during energy metabolism, and simultaneously function to modulate electrical activity via phosphorylation of voltage-gated ion channels.


Treatment advances in non-Hodgkin's lymphoma.
Non-Hodgkin's lymphomas (NHL) encompass a heterogeneous group of lymphoid malignancies with varying natural histories and prognoses. Recent classifications for NHL have defined distinct lymphoma entities based on morphology, immunophenotype, genetic features, clonal cell lineage and clinical features. These new, more precise classifications and characterizations of NHL will be essential in developing new targeted therapies. However, for this brief review, we will continue describe NHL primarily as indolent or aggressive. Treatment options for patients with indolent, but generally incurable, lymphomas include a 'watch and wait' approach, single agent alkylators, nucleoside analogues, combination chemotherapy, immunotherapy with monoclonal antibodies, radiolabelled monoclonal antibodies, or interferon (IFN). Vaccine therapy for indolent lymphomas is currently under intense investigation. For aggressive lymphomas, combination chemotherapy remains the standard of care. Major advances in the management of aggressive lymphomas include validation of the international prognostic index and clarification of the role of high-dose therapy with bone marrow or stem cell transplant in patients with relapsed aggressive lymphomas. Multiple randomised pilot trials of high dose therapy as initial therapy for aggressive lymphomas have shown conflicting results and await confirmatory studies.


Novel TRAIL sensitizer Taraxacum officinale F.H. Wigg enhances TRAIL-induced apoptosis in Huh7 cells.
TRAIL (TNF-related apoptosis inducing ligand) is a promising anti-cancer drug target that selectively induces apoptosis in cancer cells. However, many cancer cells are resistant to TRAIL-induced apoptosis. Therefore, reversing TRAIL resistance is an important step for the development of effective TRAIL-based anti-cancer therapies. We previously reported that knockdown of the TOR signaling pathway regulator-like (TIPRL) protein caused TRAIL-induced apoptosis by activation of the MKK7-c-Jun N-terminal Kinase (JNK) pathway through disruption of the MKK7-TIPRL interaction. Here, we identified Taraxacum officinale F.H. Wigg (TO) as a novel TRAIL sensitizer from a set of 500 natural products using an ELISA system and validated its activity by GST pull-down analysis. Furthermore, combination treatment of Huh7 cells with TRAIL and TO resulted in TRAIL-induced apoptosis mediated through inhibition of the MKK7-TIPRL interaction and subsequent activation of MKK7-JNK phosphorylation. Interestingly, HPLC analysis identified chicoric acid as a major component of the TO extract, and combination treatment with chicoric acid and TRAIL induced TRAIL-induced cell apoptosis via JNK activation due to inhibition of the MKK7-TIPRL interaction. Our results suggest that TO plays an important role in TRAIL-induced apoptosis, and further functional studies are warranted to confirm the importance of TO as a novel TRAIL sensitizer for cancer therapy. © 2015 Wiley Periodicals, Inc.


Type 2 diabetes: the epidemic of the new millennium.
The increasing prevalence of obesity, which has reached epidemic proportions, raises the likelihood that a similar increase in diabetes will follow. Linkage between the two conditions is clear. Overweight is not only an important risk factor for the development of diabetes, but also has a significant impact on progression and complications. Diagnostic criteria for the recognition of diabetes and for monitoring of the disease process will become increasingly important. The role of laboratory evaluation needs to be reassessed in light of new concepts regarding classification and diagnostic criteria. The relative utility of glucose and glycosylated protein measurements should be addressed, particularly the relationship between laboratory findings and clinical guidelines. Blood glucose monitoring depends on establishment of the threshold for diagnosis. Additional issues are bedside monitoring, the goal of noninvasive glucose sensors and targeting of therapy. The laboratory scientist is likely to play a key role in the application of advances in the detection and management of diabetes.


Movement prediction and movement production.
The prediction of future positions of moving objects occurs in cases of actively produced and passively observed movement. Additionally, the moving object may or may not be tracked with the eyes. The authors studied the difference between active and passive movement prediction by asking observers to estimate displacements of an occluded moving target, where the movement was driven by the observer's manual action or was passively observed. In the absence of eye tracking, they found that in the active condition, estimates are more anticipatory than in the passive conditions. Decreasing the congruence between motor action and visual feedback diminished but did not eliminate the anticipatory effect of action. When the target was tracked with the eyes, the effect of manual action disappeared. Results indicate distinct contributions of hand and eye movement signals to the prediction of trajectories of moving objects.


Inhibition of aromatase transcription via promoter II by short heterodimer partner in human preadipocytes.
Estrogen synthesis from C19 precursors is catalyzed by aromatase cytochrome P450. Overexpression of aromatase through atypical promoter usage (use of promoter II) in adipose tissue contributes to breast cancer development and progression. One tumor-derived factor that appears to contribute to this process is prostaglandin E2 (PGE2). A factor that regulates aromatase expression downstream of PGE2 is liver receptor homolog-1 (LRH-1). In a study of factors that inhibit LRH-1, we have examined the ability of short heterodimer partner (SHP) to inhibit aromatase transcription mediated by LRH-1 in preadipocytes. RT-PCR analysis indicated that both LRH-1 and SHP are expressed in human preadipocytes. To assess the effect of SHP on aromatase transcription, a transient transfection system was established using 3T3-L1 preadipocytes. Expression of SHP completely inhibited activity of an aromatase promoter II reporter gene induced by LRH-1. The combined treatment of forskolin and phorbol ester (which mimic PGE2) as well as LRH-1, which maximally induced reporter gene expression (140-fold), was also completely inhibited by SHP. This effect of SHP was mediated by inhibition of LRH-1 transcriptional activity, as measured by activity of GAL4-LRH-1 fusion constructs, and by inhibition of LRH-1 binding to promoter II. We conclude that SHP is a potent inhibitor of aromatase transcription in preadipocytes. Modulation of SHP expression and/or activity in adipose tissue may therefore have significant effects on aromatase expression and estrogen production in breast adipose tissue.


Antibiotic use in food animals: controlling the human health impact.
Resistance to antimicrobial drugs has compromised control of many bacterial pathogens. For foodborne pathogens, the most likely source of resistance is use of antimicrobials in food-producing animals. To control the human health impact from use of antimicrobials in animals, the U.S. Food and Drug Administration (FDA) recently announced plans to assess the microbial safety of all antimicrobials intended for use in food-producing animals. This paper describes the history of antimicrobial use and regulation in animals, the public health concern, the current animal drug approval process in the United States, the international perspective, and FDA's proposed procedures to evaluate the human health impact of the antimicrobial effects associated with animal drugs intended for use in food-producing animals. The primary public health goal of the improved regulatory paradigm is to ensure that significant human antimicrobial therapies are not lost due to use of antimicrobials in food animals.


Proteomic analysis reveals metabolic and regulatory systems involved in the syntrophic and axenic lifestyle of Syntrophomonas wolfei.
Microbial syntrophy is a vital metabolic interaction necessary for the complete oxidation of organic biomass to methane in all-anaerobic ecosystems. However, this process is thermodynamically constrained and represents an ecosystem-level metabolic bottleneck. To gain insight into the physiology of this process, a shotgun proteomics approach was used to quantify the protein landscape of the model syntrophic metabolizer, Syntrophomonas wolfei, grown axenically and syntrophically with Methanospirillum hungatei. Remarkably, the abundance of most proteins as represented by normalized spectral abundance factor (NSAF) value changed very little between the pure and coculture growth conditions. Among the most abundant proteins detected were GroEL and GroES chaperonins, a small heat shock protein, and proteins involved in electron transfer, beta-oxidation, and ATP synthesis. Several putative energy conservation enzyme systems that utilize NADH and ferredoxin were present. The abundance of an EtfAB2 and the membrane-bound iron-sulfur oxidoreductase (Swol_0698 gene product) delineated a potential conduit for electron transfer between acyl-CoA dehydrogenases and membrane redox carriers. Proteins detected only when S. wolfei was grown with M. hungatei included a zinc-dependent dehydrogenase with a GroES domain, whose gene is present in genomes in many organisms capable of syntrophy, and transcriptional regulators responsive to environmental stimuli or the physiological status of the cell. The proteomic analysis revealed an emphasis on macromolecular stability and energy metabolism by S. wolfei and presence of regulatory mechanisms responsive to external stimuli and cellular physiological status.


Optic nerve dysfunction during gravity inversion. Pattern reversal visual evoked potentials.
The intraocular pressure (IOP) in humans approximately doubles when a head-down, or inverted, position is assumed. Simultaneously, the ophthalmic artery pressure increases by an even greater absolute magnitude. We recorded pattern reversal visual evoked potentials (PRVEPs) in 16 subjects in both the inverted and upright positions. In ten of the 16 subjects, we performed an additional experiment. On a different day, we obtained control PRVEPs, and then, using a scleral suction cup, we artificially increased the IOPs to the same values reached when the subjects were previously inverted. We found that the PRVEP amplitudes were significantly reduced in both experiments compared with corresponding control recordings, without changes in latency or wave form. Furthermore, the degree of amplitude reduction was even greater during gravity inversion than with suction cup elevation of IOP. The effects of IOP on optic nerve function as measured by the PRVEP appear to be dependent more on the IOP than on vascular perfusion. We also concluded that gravity inversion activities pose potential risks to the eyes.


Inhibition of inducible nitric oxide synthase enhances anti-tumour immune responses in rats immunized with IFN-gamma-secreting glioma cells.
Interferon gamma (IFN-gamma) has successfully been used in immunotherapy of different experimental tumours. Mechanistically, IFN-gamma has extensive effects on the immune system including release of nitric oxide (NO) by upregulation of the inducible nitric oxide synthase (iNOS). NO has putative immunosuppressive effects but could also play a role in killing of tumour cells. Therefore, the aim of the present study was to clarify whether inhibition of iNOS in rats immunized with glioma cells (N32) producing IFN-gamma (N32-IFN-gamma), could enhance the anti-tumour immune response. Initially, both a selective iNOS, l-N(6)-(1-Iminoethyl)-l-lysine (l-NIL), and non-selective, N-nitro-l-arginine methyl ester (l-NAME), inhibitor of NOS were tested in vitro. After polyclonal stimulation with LPS and SEA, both l-NIL and l-NAME enhanced proliferation and production of IFN-gamma from activated rat splenocytes and this effect was inversely correlated to the production of NO. However, l-NIL had a broader window of efficacy and a lower minimal effective dose. When rats were immunized with N32-IFN-gamma, and administered NOS inhibitors by intraperitoneal (i.p.) mini-osmotic pumps, only splenocytes of rats treated with l-NIL, but not l-NAME, displayed an enhanced proliferation and production of IFN-gamma when re-stimulated with N32 tumour cells. Based on these findings, l-NIL was administered concurrently with N32-IFN-gamma cells to rats with intracerebral (i.c.) tumours resulting in a prolonged survival. These results show that inhibition of iNOS can enhance an IFN-gamma-based immunotherapy of experimental i.c. tumours implying that NO released after immunization has mainly immunosuppressive net effects.


Cytomegalovirus disease in a renal transplant recipient: the importance of pre-transplant screening of the donor and recipient.
A 16-year-old female patient who was born with a single kidney developed chronic kidney disease during her early childhood due to reflux nephropathy and recurrent urinary tract infection. She progressed to end-stage renal disease (ESRD) and was commenced on renal replacement therapy in the form of peritoneal dialysis in May 2011. Subsequently, she underwent living unrelated donor kidney transplantation in China. She was hospitalized soon after returning to Saudi Arabia for management of high-grade fever, shortness of breath, and deterioration of renal function, which was found to be due to cytomegalovirus (CMV) disease, proved by kidney biopsy and presence of high level of anti-CMV immunoglobulins. Allograft biopsy showed mature viral particles sized between 120 and 149 nm in the nuclei of the glomerular endothelial cells. The patient was treated with valgancyclovir and specific CMV immunoglobulin, as well as by reducing and even stopping the dose of tacrolimus and mycophenolate. Despite all these measures, her condition continued to deteriorate and she finally died. Our study emphasizes that unrelated renal transplantation, especially if unplanned and improperly prepared, is a very risky procedure that might transfer dangerous diseases and increase the morbidity and mortality of the patients. We strongly stress the need for mandatory and proper screening for CMV carrier status among donors as well as recipients prior to transplantation. Also, a recommendation is made to reject CMV-positive donors.


Protective effect of quinaprilat, an active metabolite of quinapril, on Ca2+-overload induced by lysophosphatidylcholine in isolated rat cardiomyocytes.
We examined the effects of quinaprilat, an active metabolite of quinapril (an angiotensin converting enzyme (ACE) inhibitor) on the increase in intracellular concentration of Ca2+ ([Ca2+]i) (Ca2+-overload) induced by lysophosphatidylcholine (LPC) in isolated rat cardiomyocytes. LPC (15 microM) produced Ca2+-overload with a change in cell-shape from rod to round. Quinaprilat but not quinapril at 20 or 50 microM attenuated the LPC-induced increase in [Ca2+]i and the change in cell-shape in a concentration-dependent manner. Since quinaprilat has an inhibitory action on ACE and quinapril has practically no inhibitory action on ACE, it is likely that the inhibitory action of quinaprilat on ACE is necessary for the protective effect of the drug against LPC-induced changes. We therefore examined the effects of enalapril (another ACE inhibitor with the weak inhibitory action on ACE) and enalaprilat (an active metabolite of enalapril with an inhibitory action on ACE) on the LPC-induced changes. Both enalapril and enalaprilat attenuated the LPC-induced Ca2+-overload, suggesting that the inhibitory action on ACE may not mainly contribute to the protective effect of ACE inhibitors against LPC-induced Ca2+-overload. This suggestion was supported by the fact that neither ACE (0.2 U/ml) nor angiotensin II (0.1-100 microM) increased [Ca2+]i in isolated cardiomyocytes. Furthermore, application of bradykinin (0.01-10 microM) did not enhance the protective effect of quinaprilat against LPC-induced changes. LPC also increased release of creatine kinase (CK) from the myocyte markedly, and quinaprilat but not quinapril attenuated the LPC-induced CK release. Unexpectedly, both enalapril and enalaprilat did not attenuate the LPC-induced CK release. Neither quinapril nor quinaprilat changed the critical micelle concentration of LPC, suggesting that these drugs do not directly bind to LPC. We conclude that quinaprilat attenuates the LPC-induced increase in [Ca2+]i, and that the protective effect of quinaprilat on the LPC-induced change may not be related to a decrease in angiotensin II production or an increase in bradykinin production.


Case Report: Total Nephrectomy for Renal Hydatidosis.
Cystic echinococcosis, known as hydatidosis, is a parasitic zoonosis caused by the larvae of Echinococcus granulosus. Renal hydatidosis is a very rare condition, representing 1% to 2% of cases. We present an 18-year-old patient who, after suffering a trauma, experienced severe lower back pain and persistent gross hematuria disproportionate to the trauma. Ultrasonography and tomography revealed cystic images compatible with right renal hydatidosis and hemoperitoneum. In addition, rapid clinical deterioration was observed with decreased hematocrit, leading to lumpectomy with abundant cysts inside and outside the right kidney, without viable parenchyma. Therefore, a total nephrectomy was performed. The patient was discharged with albendazole treatment. Pathological anatomic analysis confirmed the diagnosis of hydatidosis. Surgery remains the best therapeutic option. The use of ultrasonography improves the early detection of zoonosis, especially in pediatric patients, and favors the use of more conservative therapeutic techniques.


[The effects of stage based exercise program on the physical and psychological variables in stroke survivors].
The purpose of this study was to develop the exercise program based on the Transtheoretical Model, and to identify it's effects on physical and psychological variables in stroke survivors. The subjects of this study were 42 stroke survivors in the preparation and action stages. The stage based exercise program was conducted in the experimental group 8 times for 8 weeks. For data collection, physical conditioning and psychological state were measured and decisional balance and exercise specific self-efficacy were compared within groups. For the physical conditioning, there were significant improvements in shoulder's range of motion and walking speed. For the psychological state, the mean score of self esteem for the experimental group significantly increased in both stages. The pros of decisional balance increased in the preparation stage and the cons of decisional balance increased in the action stage, but the exercise specific self-efficacy scores were not significantly changed in both stages. The stage based exercise program using the Transtheoretical Model influenced on the physical and psychological variables positively for the stroke survivors. It was demonstrated that subjects became more active during the intervention.


Prediction of new low compressibility solids.
An empirical model and an ab initio calculation of the bulk moduli for covalent solids are used to suggest possible new hard materials. The empirical model indicates that hypothetical covalent solids formed between carbon and nitrogen are good candidates for extreme hardness. A prototype system is chosen and a first principles pseudopotential total energy calculation on the system is performed. The results are consistent with the empirical model and show that materials like the prototype can have bulk moduli comparable to or greater than diamond. It may be possible to synthesize such materials in the laboratory.


A self-deploying drug release device using polymeric films.
Herein, we report a sheet-type device capable of self-deployment and sustained release of protein type drugs. The device consisted of a thin photopolymerized polyethylene glycol dimethacrylate (PEGDM) sheet and collagen microparticles (COLs), which were embedded in the sheet as drug carriers and for increased drug permeation. When the density of the COLs in the sheet was increased to be sufficiently interconnected, the drug permeability was increased. In addition, since protein type drugs electrostatically interacted with the COLs, a prolonged sustained release was possible. The PEGDM/COLs device was flexible enough to be rolled up, and the device maintained its structure due to van der Waals attractive forces between the sheet surfaces. When the device was immersed in water, the attractive forces acting between the sheet surfaces were relieved by water. Subsequently, the device unfolded by bending-stress relaxation. Moreover, the rolled-up device could be injected through a conventional syringe needle into water to recover its original shape. The developed sheet-type device provides the possibility of minimally invasive transplantation into diseased tissues and organs, and could provide better therapeutic outcomes and reduce possible side effects. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 780-786, 2018.


Ultraviolet germicidal irradiation and its effects on elemental distributions in mouse embryonic fibroblast cells in x-ray fluorescence microanalysis.
Rapidly-frozen hydrated (cryopreserved) specimens combined with cryo-scanning x-ray fluorescence microscopy provide an ideal approach for investigating elemental distributions in biological cells and tissues. However, because cryopreservation does not deactivate potentially infectious agents associated with Risk Group 2 biological materials, one must be concerned with contamination of expensive and complicated cryogenic x-ray microscopes when working with such materials. We employed ultraviolet germicidal irradiation to decontaminate previously cryopreserved cells under liquid nitrogen, and then investigated its effects on elemental distributions under both frozen hydrated and freeze dried states with x-ray fluorescence microscopy. We show that the contents and distributions of most biologically important elements remain nearly unchanged when compared with non-ultraviolet-irradiated counterparts, even after multiple cycles of ultraviolet germicidal irradiation and cryogenic x-ray imaging. This provides a potential pathway for rendering Risk Group 2 biological materials safe for handling in multiuser cryogenic x-ray microscopes without affecting the fidelity of the results.


Marfan syndrome--current medical and genetic knowledge: how to treat and when.
Marfan syndrome (MFS), first described 100 years ago, remains a clinical diagnosis. Two out of 3 major systems (ocular, cardiac, skeletal) must be classically affected, to avoid overdiagnosis. Diagnosis may be confirmed by linkage to the dominantly inherited gene MFS-1 on 15q21, or by discovering the family mutation. Either technique may be used for prenatal diagnosis. Modern medical and surgical management is prolonging life, by on average, 13 years, with postoperative 20 year survival rate 65%. Beta-blocker therapy slows aortic root dilatation, and elective surgery is offered at ascending aorta diameter > or = 5 cm. Known associations with early death include new mutation, family history of dissection < 5 cm, male sex, and emergency surgery where the death rate is 5 times higher than in elective surgery. Pregnancy bears a 1% risk of fatal complication, and this risk rises with increasing aortic root diameter. Caesarean section at 38 weeks gestation should be offered if aortic root diameter is greater than 4.5 cm. Gene mapping reveals almost every mutation to be unique, interfering with multimerization of fibrillin monomers, or interactions with other connective tissue elements. Neonatal MFS is caused by mutations in exons 24-32, in calcium-binding EGF-like sequences. Mutations affecting cysteines or amino acids critical for calcium binding in other EGF-like domains, or deletions of complete EGF-like domains, generally lead to classical MFS phenotypes, as do mutations in TGF receptor-like domains. Heterogeneity has been reported with a second locus (MFS-2) on chromosome 3 in one French pedigree.


New area detector for atomic-resolution scanning transmission electron microscopy.
A new area detector for atomic-resolution scanning transmission electron microscopy (STEM) is developed and tested. The circular detector is divided into 16 segments which are individually optically coupled with photomultiplier tubes. Thus, 16 atomic-resolution STEM images which are sensitive to the spatial distribution of scattered electrons on the detector plane can be simultaneously obtained. This new detector can be potentially used not only for the simultaneous formation of common bright-field, low-angle annular dark-field and high-angle annular dark-field images, but also for the quantification of images by detecting the full range of scattered electrons and even for exploring novel atomic-resolution imaging modes by post-processing combination of the individual images.


Effects of clinical local factors on thickness and morphology of Schneiderian membrane: A retrospective clinical study.
It was informed that the thickness of maxillary sinus membrane may be affected by the local and patient-related factors in the literature. The aim of this retrospective study is to evaluate factors that may have an influence of the thickness and morphology of the maxillary sinus membrane. A total of 414 cone beam computed tomography images of 207 patients were evaluated. Radiographic parameters were evaluated at each maxillary premolar and molar tooth regions. Statistical analysis was performed to assess the association between the maxillary sinus mucosa thickness and morphology. The following factors including gender, periodontal bone loss (PBL), bone density, amount of residual alveolar bone at the edentulous space, vitality of the teeth present, and anatomical relationship between the sinus floor and posterior teeth were evaluated. The mean thickness of maxillary sinus membrane ranged between 1.47 to 2.92 mm and was significantly thicker in male subjects (P < .05). Positive correlation was detected between the sinus membrane thickness values in each posterior tooth region (P < .05). Thickening of the membrane was noted in 53% of the scans. The most commonly observed morphological change was flat thickening of the membrane (21%). No correlation was found between the evaluated local factors on the thickness and morphology of the maxillary sinus membrane (P > .05). Tooth vitality, residual alveolar bone height, and PBL seem to have no effect on the thickness and morphology of the maxillary sinus membrane.


Bilastine 10 and 20 mg in paediatric and adult patients: an updated practical approach to treatment decisions.
Bilastine, a non-sedating H1-antihistamine, is indicated to treat the symptoms of allergic disorders (e.g. rhinoconjunctivitis and urticaria) in adults and adolescents and, more recently, in children. Following its marketing approval, many questions regarding the ideal use of bilastine in various clinical practice situations have been received by the Medical Information Department (MID) of Faes Farma Spain. This article is an update of a previous review, with a focus on recent clinical information on the use of bilastine in paediatric and other populations. Results of recent clinical studies in paediatric and other populations as well as questions received and responses provided by the Faes Farma MID. The information regarding the use of bilastine in paediatric patients is the most relevant aspect of this updated review. The stepwise approval of the paediatric formulations in various countries started with the European Medicines Agency approval in 2017 in accordance with a 2009 Paediatric Investigation Plan, followed by approval in other countries. The queries that are most commonly received by the Faes Farma MID include the potential for drug interactions involving bilastine and other frequently used drugs, and the use of bilastine in special populations or to treat specific symptoms related to allergic conditions. As the concomitant use of many medications is not permitted during clinical trials, the advice provided regarding the concomitant use of other medications with bilastine considers the pharmacological properties of both the drug in question and bilastine, as well as expert opinion. Likewise, advice regarding the use of bilastine in special populations (e.g. patients with renal impairment, obesity, lactose intolerance, and elderly or pregnant individuals) or to treat specific symptoms (e.g. treatment-resistant urticaria, pruritus or BASCULE syndrome) considers the best evidence from a variety of sources, including clinical studies, real-world experience, guideline recommendations and expert opinion. This updated review provides current data regarding the best use of bilastine in specific situations and patients and identifies areas in which further knowledge is required. Although decisions regarding the use of bilastine may be aided by expert opinion that relies on knowledge of the underlying science, additional research and evidence are required to answer certain queries regarding the use of bilastine.


Photometric reconstruction of a dynamic textured surface from just one color image acquisition.
Textured surface analysis is essential for many applications. We present a three-dimensional recovery approach for real textured surfaces based on photometric stereo. The aim is to be able to measure the textured surfaces with a high degree of accuracy. For this, we use a color digital sensor and principles of color photometric stereo. This method uses a single color image, instead of a sequence of gray-scale images, to recover the surface of the three dimensions. It can thus be integrated into dynamic systems where there is significant relative motion between the object and the camera. To evaluate the performance of our method, we compare it on real textured surfaces to traditional photometric stereo using three images. We thus show that it is possible to have similar results with just one color image.


Haloxon-induced delayed neurotoxicity: effect of plasma A (aryl) esterase activity on severity of lesions in sheep.
A markedly different response to the delayed neurotoxic effects of a single dose of 400 or 800 mg/kg of the organophosphorus anthelmintic haloxon was observed in two populations of sheep. Animals with a gentically determined low level of activity of the plasma enzyme A (aryl) esterase developed clinical signs of delayed neurotoxicity within one month. Lesions relating to degeneration of myelinated nerve fibers were seen in brain stem, spinal cord, and on occasion peripheral nerve. The incidence of clinical signs, and severity of lesions as determined by semiquantitative morphological study of the spinal cord, were greater in animals given the higher dose. Neither clinical signs nor lesions relating to organophosphate-induced delayed neurotoxicity were seen in sheep with high plasma activity of A esterase.


QSAR models for binding of estrogenic compounds to estrogen receptor alpha and beta subtypes.
We have developed Quantitative Structure-Activity Relationship (QSAR) models based on Comparative Molecular Field Analysis (CoMFA) for 31 estrogenic chemicals whose relative binding affinity (RBA) is available for both ER-alpha and ER-beta. The models demonstrated a significant correlation (r2>0.95) between the CoMFA-calculated steric/electrostatic fields and corresponding RBA data and a good predictive capability (q2>0.6) based on cross-validation. The CoMFA models and contour plots obtained for ER-alpha and ER-beta suggest a close similarity between the receptors in terms of mode of binding and provide a rational basis for ligand selectivity.


The novel functions of cGMP-specific phosphodiesterase 5 and its inhibitors in carcinoma cells and pulmonary/cardiovascular vessels.
PDE5 is a key enzyme involved in the regulation of cGMP-specific signaling pathways in normal physiological processes such as smooth muscle contraction and relaxation. For this reason, inhibition of the enzyme can alter those pathophysiological conditions associated with a lowering cGMP level in tissues. For example, selective PDE5 inhibitors, such as sildenafil (Viagra, Pfizer), tadalafil (Cialis, Lilly-ICOS), and vardenafil (Levitra, Bayer), have been successfully used to treat the condition of human erectile dysfunction. More recently, the involvement of this enzyme has been proposed to influence antiproliferation and proapoptotic mechanism in multiple carcinomas. The data supporting this idea is based on increases in PDE5 activities in many carcinomas and the ability of PDE5 inhibitors such as exisulind and its analogs related to anticancer activities. Inhibition of PDE5 that results in sustained increases in [cGMP](i) are required to modify the process of apoptosis and mitotic arrest in those carcinoma cells with enhanced PDE5 expressions. Increases in PDE5 are also involved in contributing to the pathological changes in the pulmonary system resulting in hyper-proliferative remodeling of both smooth muscle and endothelium in models of pulmonary hypertension. For this reason, the use of PDE5 inhibitors in the treatment of human pulmonary hypertension has met with some success. The differences that we have previously noted in PDE isoenzymes in pulmonary arterial and microvascular endothelial cells may provide a more selective cellular strategy for use of such inhibitor. Additional studies on structure biology of these enzymes should lead to the development of agents with better cellular specificity than currently available drugs. Considering the enormous progress that has been made in the last few years, the future looks promising for agents affecting this enzyme and related systems.


Why Do Liquids Mix? The Mixing of Protic Ionic Liquids Sharing the Same Cation Is Apparently Driven by Enthalpy, Not Entropy.
We study hydrogen bond (HB) redistribution in mixtures of two protic ionic liquids (PILs) sharing the same cation: triethylammonium methanesulfonate ([TEA][OMs]) and triethylammonium trifluoromethanesulfonate ([TEA][OTf]). The mixtures exhibit large negative energies of mixing. Based on results obtained from atomic detail molecular dynamics (MD) simulations, we derive a lattice model, discriminating between HB and nonspecific intermolecular interactions. We demonstrate that due to the ordered structure of the PILs, mostly the HB interactions contribute to the mixing energy. This allows to us to connect the equilibrium of HBs to each of the two anion species with the corresponding excess energies and entropies. The entropy associated with HB redistribution is shown to be negative, and even overcompensating the positive entropy associated with a statistical distribution of the ions in the mixture. This is strongly suggesting that the mixing process is driven by enthalpy, not entropy.


Effects of fatty acids on cardioprotection by pre-ischaemic inhibition of the malate-aspartate shuttle.
1. The malate-aspartate shuttle (MAS) is the main pathway for balancing extra- and intramitochondrial glucose metabolism. Pre-ischaemic shutdown of the MAS by aminooxyacetate (AOA) mimics ischaemic preconditioning (IPC) in rat glucose-perfused hearts. The aim of the present study was to determine the effects of fatty acids (FA) on cardioprotection by pre-ischaemic inhibition of the MAS. 2. Isolated rat hearts were divided into four groups (control; pre-ischaemic AOA (0.2 mmol/L); IPC; and AOA + IPC) and were perfused with 11 mmol/L glucose, 3% bovine serum albumin plus 0, 0.4 or 1.2 mmol/L FA. The perfusion protocol included 30 min global no-flow ischaemia and 120 min reperfusion. Infarct size (IS), haemodynamic recovery, glucose oxidation and lactate release were evaluated in all four groups. 3. Pre-ischaemic AOA reduced the IS of the left ventricle in hearts perfused with 0, 0.4 and 1.2 mmol/L FA compared with that in control hearts (26 ± 2% vs 53 ± 4%, 29 ± 3% vs 53 ± 4% and 61 ± 4% vs 81 ± 3%, respectively; P < 0.01 for all). After 2 h reperfusion, AOA improved haemodynamic recovery in the absence (52 ± 2 vs 27 ± 3 mmHg in the AOA and control groups, respectively; P < 0.001) but not in the presence, of FA. Both IPC and AOA + IPC reduced IS and improved haemodynamic recovery regardless of FA levels. Postischaemic glucose oxidation was suppressed by FA and did not differ significantly between the different groups. 4. In conclusion, the reduction in IS induced by pre-ischaemic MAS shutdown is not compromised by physiological FA concentrations. Transient MAS shutdown may be involved in IPC, but is not sufficient on its own as the underlying mechanism for IPC.


Synergistic interactions of polyphenols and their effect on antiradical potential.
The aim of the current study was to evaluate interactions among polyphenols from different plants and their effect on antioxidant potential. Different mixtures of plant extracts of Crataegus oxyacantha (C), Elettaria cardamomum (Cr), Terminalia arjuna (T) and Rauvolfia serpentina (R) were prepared and evaluated for total phenolics, flavonoid contents, and antioxidant activity. A correlation was also established between total phenolics, flavonoids and antioxidant activity. Comparative evaluation revealed that phenolics, flavonoids and antioxidant activity were found high in plant extracts mixtures than individual plants. Highest phenolics (580±1.12mg GAE/g), flavonoids (67.10±0.11mg CE/g) and antioxidant activity (IC50 0.109mg/ml) was observed with ratio 1:1:1:2 of plant mixture C, Cr, T, R. A weak linear positive correlation was found between antioxidant activity, total phenolic and flavonoid contents. A negative correlation was observed among IC50 value, total phenolics and flavonoid contents. Investigation through RP-HPLC revealed the presence of different potent phenolics in plants understudy. More antioxidant potential of extracts in combinations as compared to that of individual plants was clear corroboration of synergism. The ratio (1:1:1:2) of the studied plants in combination, that showed the highest free radical potential, was another expected better pharmacological prospect. This formulation can bring maximum relief against free radical-associated diseases.


The Implantable Collamer Lens with a central port: review of the literature.
The purpose of this review is to summarize preclinical and clinical data from publications appearing in the peer-reviewed scientific literature relevant to the safety and effectiveness of the EVO Implantable Collamer Lens (ICL) posterior chamber phakic refractive lens with a central port (V4c Visian ICL with KS Aquaport, STAAR Surgical, Inc.). A literature search was conducted using PubMed.gov to identify all articles relating to the EVO ICL. Articles were examined for their relevance, and the references cited in each article were also searched for additional relevant publications. On the basis of a total of 67 preclinical studies and clinical reports, including effectiveness data on 1,905 eyes with average weighted follow-up of 12.5 months and safety data on 4,196 eyes with weighted average follow up of 14.0 months, the EVO ICL is safe and effective for the correction of a broad range of refractive errors. High levels of postoperative uncorrected visual acuity, refractive predictability, and stability demonstrate the effectiveness of the EVO ICL. Safety data suggest reduced rates of anterior subcapsular cataract and pupillary block compared with earlier models. Improved safety and proven effectiveness make EVO an attractive option for surgeons and patients.


[Extragonadal germ tumor].
Extragonadal primary tumors are uncommon and account for 3 to 5% of all germ cells neoplasias. Preferentially found in middle line structures, such tumors are frequently in an advanced stage when diagnosed. Differentiation between primary extragonadal tumors and metastasis of testicular primary tumors is essential, but may be difficult in many instances.


The Na(+)-F(1)F(0)-ATPase operon from Acetobacterium woodii. Operon structure and presence of multiple copies of atpE which encode proteolipids of 8- and 18-kda.
Eight genes (atpI, atpB, atpE(1), atpE(2), atpE(3), atpF, atpH, and atpA) upstream of and contiguous with the previously described genes atpG, atpD, and atpC were cloned from chromosomal DNA of Acetobacterium woodii. Northern blot analysis revealed that the eleven atp genes are transcribed as a polycistronic message. The atp operon encodes the Na(+)-F(1)F(0)-ATPase of A. woodii, as evident from a comparison of the biochemically derived N termini of the subunits with the amino acid sequences deduced from the DNA sequences. The molecular analysis revealed that all of the F(1)F(0)-encoding genes from Escherichia coli have homologs in the Na(+)-F(1)F(0)-ATPase operon from A. woodii, despite the fact that only six subunits were found in previous preparations of the enzyme from A. woodii. These results unequivocally prove that the Na(+)-ATPase from A. woodii is an enzyme of the F(1)F(0) class. Most interestingly, the gene encoding the proteolipid underwent quadruplication. Two gene copies (atpE(2) and atpE(3)) encode identical 8-kDa proteolipids. Two additional gene copies were fused to form the atpE(1) gene. Heterologous expression experiments as well as immunolabeling studies with native membranes revealed that atpE(1) encodes a duplicated 18-kDa proteolipid. This is the first demonstration of multiplication and fusion of proteolipid-encoding genes in F(1)F(0)-ATPase operons. Furthermore, AtpE(1) is the first duplicated proteolipid ever found to be encoded by an F(1)F(0)-ATPase operon.


Late-Onset Encephalopathy Associated With SARS-CoV-2 Infection.
Several typical and atypical neurological manifestations of viral pandemics have been reported. Neurological manifestations of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have recently been reported. In this case report, we present a patient with encephalopathy as a late neurologic manifestation of SARS-CoV-2 infection. The patient initially tested positive for the novel coronavirus after presenting with fever, cough, and altered mental status. The symptoms resolved within 5 - 7 days and the patient was discharged home. He subsequently developed worsening encephalopathy in the absence of respiratory symptoms, required hospitalization, and tested positive for SARS-CoV-2. Complete workup was unrevealing otherwise. We advise clinicians to be aware of late neurological manifestations of coronavirus disease 2019 (COVID-19) including encephalopathy.


Macrophage migration inhibitory factor counter-regulates dexamethasone-induced annexin 1 expression and influences the release of eicosanoids in murine macrophages.
Macrophage migration inhibitory factor (MIF), a pro-inflammatory cytokine and glucocorticoid (GC) counter-regulator, has emerged as an important modulator of inflammatory responses. However, the molecular mechanisms of MIF counter-regulation of GC still remain incomplete. In the present study, we investigated whether MIF mediated the counter-regulation of the anti-inflammatory effect of GC by affecting annexin 1 in RAW 264.7 macrophages. We found that stimulation of RAW 264.7 macrophages with lipopolysaccharide (LPS) resulted in down-regulation of annexin 1, while GC dexamethasone (Dex) or Dex plus LPS led to significant up-regulation of annexin 1 expression. RNA interference-mediated knockdown of intracellular MIF increased annexin 1 expression with or without incubation of Dex, whereas Dex-induced annexin 1 expression was counter-regulated by the exogenous application of recombinant MIF. Moreover, recombinant MIF counter-regulated, in a dose-dependent manner, inhibition of cytosolic phospholipase A2α (cPLA2α) activation and prostaglandin E2 (PGE2 ) and leukotriene B4 (LTB4 ) release by Dex in RAW 264.7 macrophages stimulated with LPS. Endogenous depletion of MIF enhanced the effects of Dex, reflected by further decease of cPLA2α expression and lower PGE2 and LTB4 release in RAW 264.7 macrophages. Based on these data, we suggest that MIF counter-regulates Dex-induced annexin 1 expression, further influencing the activation of cPLA2α and the release of eicosanoids. These findings will add new insights into the mechanisms of MIF counter-regulation of GC.


Stress-induced expression of p53 target genes is insensitive to SNW1/SKIP downregulation.
Pharmacological inhibition of protein kinases that are responsible for the phosphorylation of the carboxy-terminal domain (CTD) of RNA Pol II during transcription by 5,6-dichloro-1-beta-D-ribofuranosyl-benzimidazole (DRB) leads to severe inhibition of mRNA synthesis and activates p53. Transcription of the p53 effectors that are induced under these conditions, such as p21 or PUMA, must bypass the requirement for CTD phosphorylation by the positive elongation factor P-TEFb. Here, we have downregulated SNW1/SKIP, a splicing factor and a transcriptional co-regulator, which was found to interact with P-TEFb and synergistically affect Tat-dependent transcription elongation of HIV 1. Using the colon cancer derived cell line HCT116, we have found that both doxorubicin- and DRB-induced expression of p21 or PUMA is insensitive to SNW1 downregulation by siRNA. This suggests that transcription of stress response genes, unlike, e.g., the SNW1-sensitive mitosis-specific genes, can proceed uncoupled from regulators that normally function under physiological conditions.


Goldmann Applanation Tonometry versus Dynamic Contour Tonometry after Vitrectomy with Silicone Oil Endotamponade.
To evaluate the agreement of intraocular pressure (IOP) measurements using dynamic contour tonometry (DCT) and Goldmann applanation tonometry (GAT) in eyes after vitrectomy with silicone oil endotamponade and controls. In this prospective comparative study, IOP was measured with GAT and DCT in 30 eyes with oil endotamponade 1-3 days after vitrectomy and 40 untreated controls. In addition, ocular pulse amplitude (OPA), corneal pachymetry (CCT), and axial length (AL) were measured. GAT values in the oil group were significantly higher compared to control eyes (mean GAT oil 13.6 ± 5.1 mmHg; mean GAT control 10.8 ± 2.1 mmHg; p = 0.003). There was no significant difference in DCT measurements (mean DCT oil 12.0 ± 4.1 mmHg; mean DCT control 11.9 ± 2.9 mmHg; p = 0.9). This led to a significant difference of GAT-DCT between the oil and control group (mean difference of GAT-DCT oil 1.6 ± 4.7 mmHg; mean difference of GAT-DCT control -1.1 ± 2.6 mmHg; p = 0.004). The difference between GAT and DCT was negatively correlated with the mean IOP measured by both methods (r = -0.36, p = 0.02) and positively correlated with CCT only in the control group (r = 0.36, p = 0.02), as well as to AL only in the oil group (r = 0.46, p = 0.01). The OPA did not differ significantly between groups. GAT and DCT showed a good agreement in control eyes. The difference of GAT and DCT is significantly changed in eyes after vitrectomy with silicone oil endotamponade. Our findings suggest that GAT overestimates IOP in this situation.


A novel dual AMPK activator/mTOR inhibitor inhibits thyroid cancer cell growth.
Activated AMP protein kinase (AMPK) is a key regulator of intracellular energy homeostasis and may also function as a tumor suppressor by inhibiting cell growth through suppression of mammalian target of rapamycin (mTOR)/p70S6K signaling. AMPK activating agents, such as metformin and 5-aminoimidazole-4-carboxamide-ribonucleoside, have been demonstrated to inhibit thyroid cancer cell growth in in vitro and in vivo models. OSU-53, a recently developed AMPK activator, was previously shown to exhibit both in vitro and in vivo antitumor activity against aggressive breast cancer cell lines and their xenografts in nude mice. The objective of the study was to assess the in vitro effects of OSU-53 treatment in a panel of thyroid cancer cells. Experiments were performed to determine the effects of OSU-53 on cell growth, oncogenic signaling, apoptosis, autophagy, and cell rescue after selective knockdown of AMPK. OSU-53 inhibited in vitro cell growth of all seven thyroid cancer cells tested and induced activation of AMPK. Cell lines with activating mutations in RAS or BRAF, compared with cells with phosphatase and tensin homolog deleted from chromosome 10 null and RET/papillary thyroid carcinoma mutations, were more sensitive to drug treatment and demonstrated a more robust AMPK activation, inhibition of mTOR signaling, and autophagy stimulation. After selective knockdown of AMPK, cell rescue from OSU-53 treatment was not observed. We demonstrated an off-target effect of direct mTOR inhibition by OSU-53. Increased autophagy was observed in cells with activation RAS or BRAF mutations. OSU-53, a novel dual-AMPK activator/mTOR inhibitor, effectively inhibits growth in a variety of thyroid cancer cell lines and is most potent in cells with activating mutations in RAS or BRAF.


Infantile-onset inflammatory bowel disease in a patient with Hermansky-Pudlak syndrome: a case report.
Hermansky-Pudlak syndrome (HPS) is a rare, genetically heterogeneous disorder that manifests oculocutaneous albinism together with bleeding diatheses that reflect a platelet storage pool deficiency. Ten genetic subtypes of this autosomal recessive condition have been described to date. Some patients with Hermansky-Pudlak syndrome type 1, 4, or 6 develop Crohn's-like inflammatory bowel disease at any age including early childhood, but most often in adolescence or young adulthood. Here we report infantile-onset of inflammatory bowel disease in a patient with Hermansky-Pudlak syndrome type 1 who responded to infliximab. A Japanese boy, the second child of non-consanguineous healthy parents, was born with chalky white skin, silvery-white hair, and gray eyes, representing oculocutaneous albinism. He developed frequent diarrhea and fever accompanied by weight loss at 6 months, and was diagnosed with Crohn's-like inflammatory bowel disease based on the endoscopic finding of longitudinal ulcerations in the colon and the histopathologic finding of nonspecific chronic inflammation without granulomas at the age of 11 months. Treatment with an elemental diet, salazosulfapyridine, and corticosteroids failed to improve clinical or laboratory abnormalities, and the diarrhea became bloody. At 13 months he began treatment with infliximab, which produced marked improvement followed by clinical remission. Endoscopy at 20 months demonstrated healing of the colonic mucosa. At 22 months he is in sustained clinical remission receiving only infliximab. Because albinism with inflammatory bowel disease suggested Hermansky-Pudlak syndrome, we performed genetic screening using next-generation sequencing in a targeted gene panel analysis for primary immunodeficiency disease and/or inflammatory bowel disease. The patient proved to have a compound heterozygous mutation of the HPS1 gene resulting in Hermansky-Pudlak syndrome type 1. We consider this report to be the first account of type 1 Hermansky-Pudlak syndrome with infantile-onset of inflammatory bowel disease. Early administration of infliximab was effective. We recommend next-generation sequencing for patients with very early-onset inflammatory bowel disease suspected to be monogenic.


Activation of human matrix metalloproteinases by various bacterial proteinases.
Matrix metalloproteinases (MMPs) are zinc-containing proteinases that participate in tissue remodeling under physiological and pathological conditions. To test the involvement of bacterial proteinases in tissue injury during bacterial infections, we investigated the activation potential of various bacterial proteinases against precursors of MMPs (proMMPs) purified from human neutrophils (proMMP-8 and -9) and from human fibrosarcoma cells (proMMP-1). Each proMMP was subjected to treatment with a series of bacterial proteinases at molar ratios of 0.01-0.1 (bacterial proteinase to proMMP), and activities of MMPs generated were determined. Among six different bacterial proteinases, thermolysin family enzymes (family M4) such as Pseudomonas aeruginosa elastase, Vibrio cholerae proteinase, and thermolysin strongly activated all three proMMPs via limited proteolysis to generate active forms of the MMPs. N-terminal sequence analysis of the active MMPs revealed that cleavage occurred at the Val82-Leu83 and Thr90-Phe91 bonds of proMMP-1 and proMMP-9, respectively, which are located near the N terminus of the catalytic domain of MMPs. In contrast, Serratia 56-kDa proteinase and Pseudomonas alkaline proteinase, both of which are classified as members of the serralysin subfamily of zinc metalloproteinases (family M10), and Serratia 73-kDa thiol proteinase did not evidence proteolytic processing or activation of proMMP-1, -8, and -9 under these experimental conditions. These results indicate that bacterial proteinases may play an important role in tissue destruction and disintegration of extracellular matrix at the site of infections.


Associations between pre-pregnancy BMI, gestational weight gain, and prenatal diet quality in a national sample.
This secondary analysis explored the association between gestational weight gain, pre-pregnancy body mass index (BMI), and prenatal diet quality in a United States national sample. The sample comprised 1322 pregnant women in the longitudinal Infant Feeding Practices Study II with Diet History Questionnaire data. Diet quality in the third trimester was assessed using the Alternative Healthy Eating Index for Pregnancy. Self-reported pre-pregnancy BMI (categorized as underweight<18.5, normal weight 18.5-24.9, overweight 25.0-29.9, and obese≥30.0) and total gestational weight gain were used to categorize adherence to the Institute of Medicine's recommendations as inadequate, adequate, or excessive weight gain. Diet quality in pre-pregnancy BMI and gestational weight gain groups were compared using Tukey-adjusted generalized linear models adjusted for sociodemographic factors, Women, Infants, and Children participation, parity, and energy intake. Due to missing gestational weight gain data, sensitivity analyses with multiply imputed data were conducted. Women were on average 28.9 years old and of higher socioeconomic status (40% college graduates) and mostly non-Hispanic White (84%), and the mean Alternative Healthy Eating Index for Pregnancy score was 61.2 (of 130). Both pre-pregnancy BMI and gestational weight gain were inversely associated with diet quality scores (p<0.01). The interaction between pre-pregnancy BMI and gestational weight gain was significant (p = 0.04), therefore gestational weight gain models were stratified by BMI group. In stratified adjusted models, gestational weight gain was differently associated with diet quality scores (p<0.05) among women with underweight, normal weight, overweight, and obesity. The relationship between gestational weight gain and prenatal diet quality depended on pre-pregnancy BMI. For example, within women with normal weight, higher diet quality was observed in the adequate gestational weight gain group. Interventions to broadly improve prenatal diet quality are needed, however, resources can be used to target women with higher pre-pregnancy BMIs and women with inadequate or excessive gestational weight gain.


Conyzicola lurida gen. nov., sp. nov., isolated from the root of Conyza canadensis.
A novel Gram-stain-positive, non-spore-forming, pale yellow, irregular rod-shaped bacterium designated strain HWE2-01(T) was isolated from the surface-sterilized root of horseweed (Conyza canadensis). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain HWE2-01(T) belongs to the family Microbacteriaceae and showed sequence similarity levels of 97.1-97.7% with species of the genus Salinibacterium, 95.9-97.6% with species of the genus Leifsonia and 97.1% with Homoserinimonas aerilata. The highest sequence similarity (97.7%) was with Salinibacterium xinjiangense 0543(T). The genomic DNA G+C content of the novel strain was 68.1 mol%. The predominant cellular fatty acid of strain HWE2-01(T) was anteiso-C15 : 0, major menaquinones were MK-10, MK-9 and MK-11, and diagnostic polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The peptidoglycan of the novel strain contained 2,4-diaminobutyric acid, alanine, glycine and glutamic acid. The cell-wall sugars of strain HWE2-01(T) were galactose, mannose and rhamnose. The murein was of the acetyl type. Based on the results of the phenotypic and phylogenetic analysis, strain HWE2-01(T) differed in some respects from other members of the family Microbacteriaceae. Therefore, strain HWE2-01(T) is proposed to represent a novel species of a new genus in the family Microbacteriaceae with the name Conyzicola lurida gen. nov., sp. nov. (type strain = HWE2-01(T) = KCTC 29231(T) = JCM 19257(T)).


[Cryptococcosis in Hospital Carlos Van Buren, Valparaiso: a clinical serie].
Cryptococcosis is a severe opportunistic mycotic infection, caused mainly by Cryptococcus neoformans. It can present as meningitis, pneumonia or cryptococcemia. To characterize patients with Cryptococcus infection between January 1°, 2013 and June 30, 2016, in Hospital Carlos van Buren, Valparaíso, Chile. We identified retrospectively those cultures with Cryptococcus sp. growth, and then obtained their clinical files which were analyzed by two independent reviewers. We were able to obtain data from 13 of 15 patients who presented with Cryptococcus neoformans infection. Out of all, 11 (84.6%) were males, with a median age of 35 years old. 11 (84,6%) were HIV positive, 1 (7,7%) had chronic lymphocytic leukemia, and 1 (7,7%) refered alcohol abuse. Out of the 15 episodes, 9 (60%) had meningeal infection; 5 (33.3%) were cryptococcemia without meningeal involvement and 1 (6.6%) presented as a pulmonary infection. Eight patients were deceased at one year follow up. Cryptococcus sp. infection must be suspected in patients with cellular immunodeficiencies. Meningeal involvement is the most frequent form of clinical presentation. It still has a high mortality rate.


Rosmarinic Acid Attenuates Cadmium-Induced Nephrotoxicity via Inhibition of Oxidative Stress, Apoptosis, Inflammation and Fibrosis.
The present investigation was executed to reveal the protective mechanism of rosmarinic acid (RA) against cadmium (Cd)-induced nephrotoxicity. RA exhibited a concentration-dependent anti-apoptotic effect against CdCl2 in isolated mouse proximal tubular epithelial cells. Cd treatment significantly (p < 0.01) imparted oxidative stress to the renal cells via excessive ROS production, triggering NO level, NADPH oxidase activation, and impairment of cellular redox defense system. Cd-mediated oxidative stress significantly (p < 0.01) endorsed apoptosis to the murine kidney cells by triggering NF-κB/PKC-δ/TNFR2 activation. In addition, CdCl2 induced renal fibrosis by triggering TGF-β1/SMAD3/α-SMA/collagen signaling within renal cells. On the other hand, RA significantly (p < 0.05-0.01) attenuated Cd-provoked oxidative stress and associated pathological signal transduction in murine renal cells. RA treatment also could significantly (p < 0.05-0.01) reciprocate Cd-mediated pathological changes in blood and urine parameters in mice. In addition, histological data supported the pharmacological findings. In silico chemometric analyses predicted the possible interactions between RA and different signal proteins and anticipated drug-likeness characteristics of RA. Hence, RA can potentially be applied as a therapeutic agent to treat Cd-mediated nephrotoxicity in future.


[Studies of the expression of human interleukin-2 receptor cDNA in mammalian cells].
Human interleukin-2 receptor cDNA was transfected into mouse fibroblast cells (L929) by using DNA-calcium phosphate coprecipitation method. Results from RNA dot-blot hybri dization, FITC-IL-2 fluorescent staining assay and anti-Tac specific rossette test demonstrated that the product of human interleukin-2 receptor cDNA in L929 cells is able to bind IL-2 and anti-Tac antibody. Moreover, the abnormal expression of IL-2R gene in T-cell leukemia cell lines such as Jukat cells and Molt-4 cells was analyzed and discussed.


Enhancement of protective immune responses induced by Toxoplasma gondii dense granule antigen 7 (GRA7) against toxoplasmosis in mice using a prime-boost vaccination strategy.
Effective vaccines against Toxoplasma gondii may contribute to preventing and controlling the spread of toxoplasmosis, which is important for improving outcomes of infections in humans and livestock animals. The dense granule antigen 7 (GRA7) of T. gondii might be an immunodominant antigen for a vaccine candidate. In the present study, a further exploration of its vaccine effect, a heterologous prime-boost vaccination strategy with a recombinant eukaryotic plasmid pEGFP-GRA7 and a recombinant protein GRA7 expressed from a prokaryotic plasmid pET30-GRA7, was performed in BALB/c mice. The data reveal that a DNA prime-protein boost vaccination induces both humoral and cellular immune responses against T. gondii associated with high levels of total IgG, IgG2a isotype and gamma interferon (IFN-γ). Challenge experiments further show that the DNA prime-protein boost vaccination significantly increases survival rate (60%), compared with controls in which all died within 8 days of challenge. Therefore, the DNA prime-protein boost vaccination based on GRA7 might be a promising regimen for further development of an effective vaccine against T. gondii.


Smallpox: clinical highlights and considerations for vaccination.
Smallpox virus has gained considerable attention as a potential bioterrorism agent. Recommendations for smallpox (vaccinia) vaccination presume a low risk for use of smallpox as a terrorist biological agent and vaccination is currently recommended for selected groups of individuals such as health care workers, public health authorities, and emergency/rescue workers, among others. Information about adverse reactions to the smallpox vaccine is based upon studies completed during the 1950s and 1960s. The prevalence of various diseases has changed over the last four decades and new disease entities have been described during this period. The smallpox vaccination may be contra-indicated in many of these conditions. This has made pre-screening of potential vaccines necessary. It is believed that at present, the risks of vaccine-associated complications far outweigh the potential benefits of vaccination in the general population.


The Synergistic Effect of Biosynthesized Silver Nanoparticles and Phytocompound as a Novel Approach to the Elimination of Pathogens.
Due to the wide applications of silver nanoparticles (AgNPs), research on their ecological synthesis has been extensive in recent years. In our study, biogenic silver nanoparticles were synthesized extracellularly using the white rot fungus Trametes versicolor via two cultivation methods: static and shaking. The cell filtrate of the fungus was used as a reducing agent in the process of nanoparticle synthesis. Characterization of the obtained nanoparticles was carried out using UV-VIS spectroscopy and scanning electron microscopy. The biosynthesized nanoparticles have antimicrobial potential against pathogenic bacteria, particularly in Gram-negative strains. The bactericidal effect was obtained for E. coli at a concentration of 7 µg/mL. The use of higher concentrations of compounds was necessary for Gram-positive bacteria. Taking into account the problem of the risk of cytotoxicity of AgNPs, combined therapy using a phytochemical was used for the first time, which was aimed at reducing the doses of nanoparticles. The most representative synergistic effect was observed in the treatment of 5 µg/mL silver nanoparticles in combination with 15 µg/mL ursolic acid against E. coli and P. aeruginosa with a bactericidal effect. Moreover, the coadministration of nanoparticles considerably reduced the growth of both Staphylococcus strains, with a bactericidal effect against S. aureus. The viability test confirmed the strong synergistic effect of both tested compounds. Silver nanoparticles synthesized using the T. versicolor showed excellent antibacterial potential, which opens perspectives for future investigations concerning the use of the nanoparticles as antimicrobials in the areas of health.


Cohesin-dependent regulation of gene expression during differentiation is lost in cohesin-mutated myeloid malignancies.
Cohesin complex disruption alters gene expression, and cohesin mutations are common in myeloid neoplasia, suggesting a critical role in hematopoiesis. Here, we explore cohesin dynamics and regulation of hematopoietic stem cell homeostasis and differentiation. Cohesin binding increases at active regulatory elements only during erythroid differentiation. Prior binding of the repressive Ets transcription factor Etv6 predicts cohesin binding at these elements and Etv6 interacts with cohesin at chromatin. Depletion of cohesin severely impairs erythroid differentiation, particularly at Etv6-prebound loci, but augments self-renewal programs. Together with corroborative findings in acute myeloid leukemia and myelodysplastic syndrome patient samples, these data suggest cohesin-mediated alleviation of Etv6 repression is required for dynamic expression at critical erythroid genes during differentiation and how this may be perturbed in myeloid malignancies.


Thumb Carpometacarpal Arthritis Surgery: The Patient Experience.
Patients with symptomatic recalcitrant thumb carpometacarpal arthritis often undergo surgery. Although most surgical patients do well, the authors anticipated that a substantial portion of their thumb carpometacarpal surgery patients would have unsatisfactory experiences and express unmet expectations, dissatisfaction, and regret, regardless of surgical procedure performed. The authors hypothesized those experiences would correlate with patient-reported outcomes scores. The authors identified patients who had undergone trapeziectomy alone or with ligament reconstruction 1 to 4 years previously for primary thumb carpometacarpal arthritis. One hundred twelve patients completed Quick Disabilities of the Arm, Shoulder and Hand and visual analogue scale pain, expectations, satisfaction, and regret questionnaires. More than 40 percent of patients expected to "return to normal" after surgery for pain, strength, and/or function. Including all patients, 7, 19, and 11 percent had unmet expectations for improvement in pain, strength, and function, respectively. Twelve percent expressed dissatisfaction with their outcome. Although just 4 percent regretted undergoing surgery, 13 percent would likely not recommend the procedure to someone they care about. There were no statistically significant differences for any patient-reported outcomes between trapeziectomy-alone (n = 20) and trapeziectomy with ligament reconstruction (n = 92). Visual analogue scale and Quick Disabilities of the Arm, Shoulder and Hand questionnaire scores were both moderately correlated with expectations being met for pain, strength, and function and for satisfaction with surgical outcome. Patients' thumb carpometacarpal surgical experiences vary considerably. Many express dissatisfaction or a lack of expectations met with the two most common procedures. A thorough understanding and review of expectations preoperatively may be uniquely pertinent for these patients. Further research should determine predictors and potentially modifiable factors for unsatisfactory outcomes.


Intercomparison of methods of coupling between convection and large-scale circulation: 2. Comparison over nonuniform surface conditions.
As part of an international intercomparison project, the weak temperature gradient (WTG) and damped gravity wave (DGW) methods are used to parameterize large-scale dynamics in a set of cloud-resolving models (CRMs) and single column models (SCMs). The WTG or DGW method is implemented using a configuration that couples a model to a reference state defined with profiles obtained from the same model in radiative-convective equilibrium. We investigated the sensitivity of each model to changes in SST, given a fixed reference state. We performed a systematic comparison of the WTG and DGW methods in different models, and a systematic comparison of the behavior of those models using the WTG method and the DGW method. The sensitivity to the SST depends on both the large-scale parameterization method and the choice of the cloud model. In general, SCMs display a wider range of behaviors than CRMs. All CRMs using either the WTG or DGW method show an increase of precipitation with SST, while SCMs show sensitivities which are not always monotonic. CRMs using either the WTG or DGW method show a similar relationship between mean precipitation rate and column-relative humidity, while SCMs exhibit a much wider range of behaviors. DGW simulations produce large-scale velocity profiles which are smoother and less top-heavy compared to those produced by the WTG simulations. These large-scale parameterization methods provide a useful tool to identify the impact of parameterization differences on model behavior in the presence of two-way feedback between convection and the large-scale circulation.


Feed substrates influence biofilm formation on reverse osmosis membranes and their cleaning efficiency.
The dairy industry is increasingly using reverse osmosis (RO) membranes for concentration of various fluid feed materials such as whey and ultrafiltration (UF) permeate. This study compared the effect of UF permeate and whey on membrane biofilm formation. A Bacillus sp., previously isolated in our laboratory from a cleaning-resistant membrane biofilm, was used to develop 48-h-old static biofilms on RO membrane pieces, using the different feed substrates (UF permeate, whey, and an alternating whey/UF feed). Biofilms were analyzed for viable counts by the swab technique, and we used scanning electron and atomic force microscopy for microstructure imaging. The membrane cleaning process included 6 sequential steps. We observed differences in the resistance pattern of the 3 types of biofilms to the typical cleaning process. The mean pretreatment counts of the 48-h UF permeate biofilms were 5.39 log cfu/cm2, much higher than the whey biofilm counts of 3.44 log, and alternating whey/UF biofilm counts of 4.54 log. After a 6-step cleaning cycle, we found 2.54 log survivors of the Bacillus isolate on UF biofilms, whereas only 1.82 log survivors were found in whey biofilm, and 2.14 log survivors on whey/UF permeate biofilms. In conclusion, the UF permeate biofilms was more resistant to the biofilm cleaning process compared with the whey or whey/UF permeate biofilms. Scanning electron micrographs showed different microstructures of biofilms based on the type of feed. For UF permeate and whey/UF permeate biofilms, bacilli were present in multilayers of cells in aggregates or irregular clusters with foulant layers. In contrast, those in whey biofilms were in monolayers, with a smoother, flatter appearance. Atomic force microscopy analysis indicated that UF permeate biofilms had the greatest surface roughness among the biofilms, reflecting intensified bacterial colonization. The biofilm micro- and nanostructure variations for the 2 feed substrates and their combination may have resulted in differences in their resistance to the cleaning process.


Tricalcium phosphate (-containing) biomaterials in the treatment of periodontal infra-bony defects: A systematic review and meta-analysis.
This study aimed to investigate the additional clinical benefit of tricalcium phosphate (TCP) (-containing) biomaterials compared to open flap debridement (OFD) in periodontal infra-bony defects. A literature search was conducted in Pubmed, Embase and Cochrane library for entries published up to 14th July 2021. All randomized controlled trials (RCTs) that compared a TCP (-containing) material with OFD and studies that assessed a beta-TCP group alone, with vertical defect sites with PPD of ≥ 6 mm and/or presence of infra-bony defects of ≥ 3 mm and a minimum follow-up of 6 months were included. Risk of bias was assessed with the Oxford scale. The random-effects (RE) model was synthesized as differences between weighted average (MD) for probing pocket depth (PPD) and clinical attachment level (CAL) between TCP and OFD groups. An RE analysis was also performed for the beta-TCP group alone. Data from 16 RCTs were included in the analysis. Six studies that represented 151 patients and sites were selected for meta-analysis. The overall MD with 95% CI at 6 months was calculated to be -0.47 [-0.83, -0.12; P = 0.0087] and -1.06 [-1.67, -0.46; P = 0.0006] for PPD and CAL, respectively. Whereas MD at 12 months for PPD and CAL was -0.89 [-1.54, -0.23; P = 0.0078] and -1.25 [-1.85, -0.66; P<0.0001], respectively. All results were in favor of TCP (-containing) group over OFD. The results of the study suggest that the use of a TCP (-containing) material may have the potential for additional clinical improvement in PPD and CAL compared with OFD in infra-bony defects, given the limitations of the included evidence. The use of TCP as a bone graft substitute is becoming increasingly common. Therefore, it would be advantageous if an adjunctive benefit in the regeneration of infra-bony defects could be demonstrated to facilitate material selection.


[Contractile function of the myocardium of the right ventricle in thyrotoxicosis].
The functional condition of the myocardium of the right ventricle was studied in 98 patients suffering from thyrotoxicosis and in 28 healthy persons. The pressure in the pulmonary artery, the tension index of the myocardium and the frequency of cardiac contractions proved to increase against the background of an excess of thyroid hormones in the organism. The contractility index of the myocardium, the volume velocity of the ejection, the rate of displacement of the wall of the ventricle during blood expulsion increased at the early stages of the disease, and later became lowered. Under experimental conditions the initial period of thyoidin intoxication was characterized by increased work of the right ventricle. The electron microscopic picture corresponding to this phase of the process pointed to the injurious action of the thyroid gland hormones on the mitochondria. The greatest increase in the functional intensity of the myocardium of the right ventricle was reached on the 14th day of the experimental thyroidin intoxication. The ultrastructural changes recorded at this period reflected the growing hyperfunction of the mitochondria and the development of destructive changes in the contractile aparatus of the myocytes.


The structure of postconcussion symptoms on the Neurobehavioral Symptom Inventory: a comparison of alternative models.
To evaluate and compare the existing Neurobehavioral Symptom Inventory factor structure models to determine which model provides the best overall fit for postconcussion symptoms and determine which model is useful across different samples (eg, with and without mild traumatic brain injury [TBI] history). N/A. A Florida National Guard sample (N = 3098) and a national Department of Veterans Affairs sample (N = 48,175). Retrospective structural equation modeling was used to compare 16 alternative factor structure models. First, these 16 possible models were examined separately in both samples. Then, to determine whether the same factor structures applied across subsamples within these samples, the models were compared for those deployed and those not deployed in the Florida National Guard sample and between those with TBI confirmed on clinical evaluation and those who were determined not to have sustained a TBI within the Department of Veterans Affairs sample. Neurobehavioral Symptom Inventory. A 4-factor model--vestibular, somatic, cognitive, and affective--had the best overall fit, after elimination of 2 items (ie, hearing problems and appetite disturbance), and was most applicable across samples. These findings extend the findings of Meterko et al to other samples. Because findings were consistent across sample and subsamples, the current findings are applicable to both Department of Veteran Affairs and Department of Defense postdeployment medical evaluation settings.


Merging information in speech recognition: feedback is never necessary.
Top-down feedback does not benefit speech recognition; on the contrary, it can hinder it. No experimental data imply that feedback loops are required for speech recognition. Feedback is accordingly unnecessary and spoken word recognition is modular. To defend this thesis, we analyse lexical involvement in phonemic decision making. TRACE (McClelland & Elman 1986), a model with feedback from the lexicon to prelexical processes, is unable to account for all the available data on phonemic decision making. The modular Race model (Cutler & Norris 1979) is likewise challenged by some recent results, however. We therefore present a new modular model of phonemic decision making, the Merge model. In Merge, information flows from prelexical processes to the lexicon without feedback. Because phonemic decisions are based on the merging of prelexical and lexical information, Merge correctly predicts lexical involvement in phonemic decisions in both words and nonwords. Computer simulations show how Merge is able to account for the data through a process of competition between lexical hypotheses. We discuss the issue of feedback in other areas of language processing and conclude that modular models are particularly well suited to the problems and constraints of speech recognition.


Natural variation meets synthetic biology: Promiscuous trichome-expressed acyltransferases from Nicotiana.
Acylsugars are defensive, trichome-synthesized sugar esters produced in plants across the Solanaceae (nightshade) family. Although assembled from simple metabolites and synthesized by a relatively short core biosynthetic pathway, tremendous within- and across-species acylsugar structural variation is documented across the family. To advance our understanding of the diversity and the synthesis of acylsugars within the Nicotiana genus, trichome extracts were profiled across the genus coupled with transcriptomics-guided enzyme discovery and in vivo and in vitro analysis. Differences in the types of sugar cores, numbers of acylations, and acyl chain structures contributed to over 300 unique annotated acylsugars throughout Nicotiana. Placement of acyl chain length into a phylogenetic context revealed that an unsaturated acyl chain type was detected in a few closely related species. A comparative transcriptomics approach identified trichome-enriched Nicotiana acuminata acylsugar biosynthetic candidate enzymes. More than 25 acylsugar variants could be produced in a single enzyme assay with four N. acuminata acylsugar acyltransferases (NacASAT1-4) together with structurally diverse acyl-CoAs and sucrose. Liquid chromatography coupled with mass spectrometry screening of in vitro products revealed the ability of these enzymes to make acylsugars not present in Nicotiana plant extracts. In vitro acylsugar production also provided insights into acyltransferase acyl donor promiscuity and acyl acceptor specificity as well as regiospecificity of some ASATs. This study suggests that promiscuous Nicotiana acyltransferases can be used as synthetic biology tools to produce novel and potentially useful metabolites.