Recent advances on bioorganic chemistry of plant metabolites controlling nyctinasty.
Leguminous plants open their leaves during the daytime and close them at night as if sleeping, a type of movement that follows circadian rhythms, and is known as nyctinastic movement. This phenomenon is controlled by two endogenous bioactive substances that exhibit opposing activities: Leaf-Opening Factor (LOF), which opens the leaves, and Leaf-Closing Factor (LCF), which closes them. The authors have carried out chemical biological research using these bioactive substances as molecular probes in order to clarify the mechanisms of nyctinastic movement. Here, we report on the detection and identification of the target proteins of these compounds using original methodology.


Aggressive and Violent Behavior and Emotional Self-Efficacy: Is There a Relationship for Adolescents?
In this cross-sectional study we explored relationships between aggressive and violent behaviors and emotional self-efficacy (ESE) in a statewide sample of public high school adolescents in South Carolina (N = 3836). The US Centers for Disease Control and Prevention Youth Risk Behavior Survey items on aggressive and violent behavior items and an adolescent ESE scale were used. Logistic regression analyses and multivariate models constructed separately, revealed significant race by sex findings. Results suggest that carrying a weapon to school (past 30 days) and being threatened or injured with a gun, knife, or club at school (past 12 months) were significantly associated (p < .05) with reduced ESE for specific race/sex groups. Results have implications for school- and community-based mental health services and social and emotional learning and aggression/violence prevention programs for adolescents. Measures of ESE as a component of comprehensive assessments of adolescent mental health, social and emotional learning and aggressive/violent behaviors in fieldwork, research, and program-evaluation efforts should be considered.


Flame retardant chloroprene rubbers with high tensile strength and elongation at break via dual cross-linked networks.
The tensile strength and elongation at break of rubbers are mutually restrictive factors. Design and preparation of chloroprene rubber (CR) with high tensile strength, high elongation at break and excellent flame retardancy at the same time is challenging. Melamine cyanurate (MCA) is for the first time discovered to be a reactive flame retardant for CR. The tensile strength of C-M36 (with 3 wt% ZnO and 36 wt% MCA) vulcanizate is 2.5 times that of C-M0 (only with 3 wt% ZnO) vulcanizate, while the elongation at break of C-M36 vulcanizate is 1.3 times that of ZnO cross-linked C-M0 vulcanizate. At the same time, the limiting oxygen index of C-M36 (39%) is 1.22 times that of C-M0 (32%). FTIR and the vulcanization tests confirm that the reaction between CR and cyanuric acid occurs under the catalysis of a base (melamine), and the cyanuric acid molecules are grafted onto the molecular chain of CR. Two types of crosslinking networks are formed in CR vulcanizate, namely the traditional covalent bond crosslinks and the triple hydrogen crosslinks formed between cyanuric acid and melamine. Thus, the flame-retardant CR/MCA vulcanizate with high strength and high elongation at break is obtained. This research will strongly promote the industrial application of CR.


The agonism and synergistic potentiation of weak partial agonists by triethylamine in alpha 1-adrenergic receptor activation: evidence for a salt bridge as the initiating process.
Alpha 1-adrenergic receptor (AR) activation is thought to be initiated by disruption of a constraining interhelical salt bridge (). Disruption of this salt bridge is achieved through a competition for the aspartic acid residue in transmembrane domain three by the protonated amine of the endogenous ligand norepinephrine and a lysine residue in transmembrane domain seven. To further test this hypothesis, we investigated the possibility that a simple amine could mimic an important functional group of the endogenous ligand and break this alpha 1-AR ionic constraint leading to agonism. Triethylamine (TEA) was able to generate concentration-dependent increases of soluble inositol phosphates in COS-1 cells transiently transfected with the hamster alpha 1b-AR and in Rat-1 fibroblasts stably transfected with the human alpha 1a-AR subtype. TEA was also able to synergistically potentiate the second messenger production by weak partial alpha 1-AR agonists and this effect was fully inhibited by the alpha 1-AR antagonist prazosin. However, this synergistic potentiation was not observed for full alpha 1-AR agonists. Instead, TEA caused a parallel rightward shift of the dose-response curve, consistent with the properties of competitive antagonism. TEA specifically bound to a single population of alpha 1-ARs with a Ki of 28.7 +/- 4.7 mM. In addition, the site of binding by TEA to the alpha 1-AR is at the conserved aspartic acid residue in transmembrane domain three, which is part of the constraining salt bridge. These results indicate a direct interaction of TEA in the receptor agonist binding pocket that leads to a disruption of the constraining salt bridge, thereby initiating alpha 1-AR activation.


Hypomagnesemia inhibits nitric oxide release from coronary endothelium: protective role of magnesium infusion after cardiac operations.
Postoperative hypomagnesemia is common in patients who have undergone cardiac operations and is associated with clinically significant morbidity resulting from atrial and ventricular dysrhythmias. Magnesium supplementation may increase the cardiac index in the early postoperative period. The action of the magnesium cation on coronary vascular reactivity was studied. Segments of canine epicardial coronary artery were suspended in organ chambers to measure isometric force (95% O2/5% CO2, 37 degrees C). In coronary segments constricted with prostaglandin F2alpha (2 x 10[-6] mol/L), acetylcholine and adenosine diphosphate (10[-9] to 10[-4] mol/L) induced vasodilation in arteries with endothelium (n=10, each group; p < 0.05). Acetylcholine-mediated vasodilation was blocked by NG-monomethyl-L-arginine (10[-4] mol/L) and NG-nitro-L-arginine (10[-4] mol/L), two inhibitors of nitric oxide synthesis from L-arginine (n=10, p < 0.05). The removal of magnesium from the organ chamber solution impaired vasodilation in response to acetylcholine and adenosine diphosphate. However, normal endothelium-dependent vasodilation could be restored by return of magnesium to the bathing solution. Vascular relaxation in response to bradykinin (10[-9] to 10[-6] mol/L), which was found to induce endothelium-dependent vasodilation independent of nitric oxide production, was unaffected by magnesium removal (n=10). Hypomagnesemia selectively impaired the release of nitric oxide from the coronary endothelium. Because nitric oxide is a potent endogenous nitro-vasodilator and inhibitor of platelet aggregation and adhesion, hypomagnesemia could promote vasoconstriction and coronary thrombosis in the early postoperative period.


Use of tylosin to prevent early mortality in bobwhite quail.
Two experiments were conducted to study the effect of treating day-old quail by drenching either with a soluble tylosin, tartrate solution or plain water, or medicating for five days with tylosin in the drinking water. Diagnosis of dead birds indicated ulcerative enteritis infection. Quail drenched with plain water or tylosine had less (P less than .01) mortality than those given tylosin in the drinking water or the untreated control birds. There was no difference between quail drenched with plain water or tylosin. Birds given tylosin in the drinking water resisted mortality significantly better (P less than .01) than the control birds.


Association between age at menarche and cardiometabolic risk among Samoan adults.
Recent studies suggest that early menarche may increase cardiometabolic morbidity and mortality. Yet few studies have examined this association in the Pacific Islands, where obesity prevalence is among the highest globally. We sought to examine associations between age at menarche and cardiometabolic risk in Samoa. Participants were from the Soifua Manuia study (n = 285, age 32-72 years) conducted in Samoa from 2017 to 2019. Logistic regressions were conducted to estimate odds of obesity, hypertension, diabetes, dyslipidemia, and metabolic syndrome per one-year increase in age at menarche. Linear regressions were conducted to examine associations between age at menarche and continuous measures of adiposity, blood pressure, insulin resistance, and serum lipids. Median age at menarche was 14 years (IQR = 2). After controlling for relevant covariates, each one-year increase in age at menarche was associated with a 15% decrease (OR = 0.85, 95% CI: 0.72-1.01, p = .067) in odds of hypertension, but a 21% increase (OR = 1.21, 95% CI: 1.01-1.45, p = .044) in odds of diabetes and 18% increase (OR = 1.18, 95% CI: 0.98-1.42, p = .081) in odds of high total cholesterol. Each additional year in age at menarche was associated with a 1.60 ± 0.52 kg (p = .002) decrease in lean mass and 1.56 ± 0.51 kg (p = .003) decrease in fat-free mass. Associations between age at menarche and cardiometabolic risk may be population-specific and are likely influenced by both current and historical nutritional and epidemiological contexts. Prospective studies are needed to clarify the role of childhood adiposity and other early life exposures on age at menarche and subsequent cardiometabolic risk.


Safety and efficacy evaluation of low-dose trimethoprim-sulfamethoxazole for prophylaxis of Pneumocystis pneumonia in HIV uninfected patients undergoing hemodialysis: a retrospective observational study.
Pneumocystis pneumonia (PCP) is a potentially life-threatening infection. Trimethoprim-sulfamethoxazole (TMP-SMX) is considered as the first regimen for PCP prophylaxis according to several guidelines. The recommended prophylactic dose of TMP-SMX has been determined based on patients with normal renal function, but the appropriate dosage for patients undergoing hemodialysis is unknown. The aim of this study was to investigate the efficacy and safety of low-dose TMP-SMX in patients undergoing hemodialysis. HIV-uninfected adult patients who were undergoing hemodialysis and administered TMP-SMX for PCP prophylaxis, were included, and divided into standard-dose (≥6 single strength (SS, TMP-SMX 80 mg/400 mg tablets/week) and low-dose groups (< 6 SS tablets/week). The endpoints were cumulative incidence of PCP and cumulative discontinuation rate of TMP-SMX due to adverse events. For comparison of the groups, we employed the chi-squared test for categorical variables and the Mann-Whitney U test for continuous variables. Risk factors for the endpoints were evaluated using the Cox Fine and Gray method. The median age of the 81 patients included in the study was 67 years (IQR: 60-76 years), and 52 patients (64.2%) were men. No patients in either group developed PCP during the observation period. The yearly cumulative incidence of discontinuation was 12.1% (95% confidence interval [CI]: 0.027-0.29) in the low-dose group and 35.6% (95% CI: 0.20-0.52) in the standard-dose group (P = 0.019). The adjusted hazard ratio of the low-dose group compared to standard-dose group was 0.18 (95% CI: 0.04-0.86, P = 0.032). None of the study patients developed PCP, and the cumulative discontinuation rate of TMP-SMX due to adverse events was significantly lower in the low-dose group compared to that in the standard-dose group (P = 0.032). These results indicate that low-dose TMP-SMX is an appropriate regimen to maintain a balance between PCP prophylaxis and prevention of adverse events due to TMP-SMX administration. These findings can guide health care professionals to determine TMP-SMX dosage when considering PCP prophylaxis for patients undergoing hemodialysis.


A Place to Call Home: An Analysis of the Bacterial Communities in Two Tethya rubra Samaai and Gibbons 2005 Populations in Algoa Bay, South Africa.
Sponges are important sources of bioactive secondary metabolites. These compounds are frequently synthesized by bacterial symbionts, which may be recruited from the surrounding seawater or transferred to the sponge progeny by the parent. In this study, we investigated the bacterial communities associated with the sponge Tethya rubra Samaai and Gibbons 2005. Sponge specimens were collected from Evans Peak and RIY Banks reefs in Algoa Bay, South Africa and taxonomically identified by spicule analysis and molecular barcoding. Crude chemical extracts generated from individual sponges were profiled by ultraviolet high performance liquid chromatography (UV-HPLC) and subjected to bioactivity assays in mammalian cells. Next-generation sequencing analysis of 16S rRNA gene sequences was used to characterize sponge-associated bacterial communities. T. rubra sponges collected from the two locations were morphologically and genetically indistinguishable. Chemical extracts from sponges collected at RIY banks showed mild inhibition of the metabolic activity of mammalian cells and their UV-HPLC profiles were distinct from those of sponges collected at Evans Peak. Similarly, the bacterial communities associated with sponges from the two locations were distinct with evidence of vertical transmission of symbionts from the sponge parent to its embryos. We conclude that these distinct bacterial communities may be responsible for the differences observed in the chemical profiles of the two Algoa Bay T. rubra Samaai and Gibbons 2005 populations.


Pulmonary alveolar epithelial permeability in surgically induced hemorrhagic pancreatitis in dogs.
The adult respiratory distress syndrome has been associated with acute pancreatitis in 5-8% of patients. to determine, in a quantitative manner, the effects of acute, hemorrhagic pancreatitis on alveolar epithelial permeability, we studied two groups of dogs (group 1 - sham-operated, 7 dogs; group 2 - surgically induced pancreatitis, 7 dogs) and measured the flux of 10,000-20,000 molecular weight dextran and albumin (69,000 molecular weight), from the blood to the saline-filled lung. Following the surgical procedure (day 1), serum amylase levels increased significantly (397 +/- 70-1,268 +/- 95 dye units) in group 2 but not in group 1. Alveolar epithelial permeability (expressed as T1/2 time in minutes to 50% equilibration between blood and lung liquid) did not increase in either group for dextrans or albumin. We conclude that experimental acute pancreatitis does not cause an increase in alveolar epithelial permeability for molecular weight substances up to 69,000 daltons.


Exposure to secondhand smoke and asthma severity among children in Connecticut.
To determine whether secondhand smoke (SHS) exposure is associated with greater asthma severity in children with physician-diagnosed asthma living in CT, and to examine whether area of residence, race/ethnicity or poverty moderate the association. A large childhood asthma database in CT (Easy Breathing) was linked by participant zip code to census data to classify participants by area of residence. Multinomial logistic regression models, adjusted for enrollment date, sex, age, race/ethnicity, area of residence, insurance type, family history of asthma, eczema, and exposure to dogs, cats, gas stove, rodents and cockroaches were used to examine the association between self-reported exposure to SHS and clinician-determined asthma severity (mild, moderate, and severe persistent vs. intermittent asthma). Of the 30,163 children with asthma enrolled in Easy Breathing, between 6 months and 18 years old, living in 161 different towns in CT, exposure to SHS was associated with greater asthma severity (adjusted relative risk ratio (aRRR): 1.07 [1.00, 1.15] and aRRR: 1.11 [1.02, 1.22] for mild and moderate persistent asthma, respectively). The odds of Black and Puerto Rican/Hispanic children with asthma being exposed to SHS were twice that of Caucasian children. Though the odds of SHS exposure for publicly insured children with asthma were three times greater than the odds for privately insured children (OR: 3.02 [2.84,3,21]), SHS exposure was associated with persistent asthma only among privately insured children (adjusted odds ratio (aOR): 1.23 [1.11,1.37]). This is the first large-scale pragmatic study to demonstrate that children exposed to SHS in Connecticut have greater asthma severity, clinically determined using a systematic approach, and varies by insurance status.


Central Projection of Antennal Sensory Neurons in the Central Nervous System of the Mirid Bug Apolygus lucorum (Meyer-Dür).
The mirid bug Apolygus lucorum (Meyer-Dür), a polyphagous pest, is dependent on olfactory cues to locate various host plant species and mates. In this study, we traced the projection pathway of the antennal sensory neurons and visualized their projection patterns in the central nervous system of A. lucorum through confocal microscopy and digital reconstructions. We also examined the glomerular organization of the primary olfactory center of the brain, the antennal lobe, and created a three-dimensional model of the glomeruli. We found that the axons of the sensory neurons project into the brain via the ipsilateral antennal nerve, and descend further into the gnathal ganglion, prothoracic ganglion, mesothoracic ganglion, and metathoracic ganglion, and reach as far as to the abdominal ganglion. Such a projection pattern indicates that antennal sensory neurons of A. lucorum may be potentially directly connected to motor neurons. The antennal lobe, however, is the major target area of antennal sensory neurons. The antennal lobe is composed of a large number of glomeruli, i.e. 70-80 glomeruli in one AL of A. lucorum. The results of this study which provide information about the basic anatomical arrangement of the brain olfactory center of A. lucorum, are important for further investigations of chemosensory encoding mechanisms of the mirid bug.


Gain-of-function mutations in complement factor B are associated with atypical hemolytic uremic syndrome.
Hemolytic uremic syndrome (HUS) is an important cause of acute renal failure in children. Mutations in one or more genes encoding complement-regulatory proteins have been reported in approximately one-third of nondiarrheal, atypical HUS (aHUS) patients, suggesting a defect in the protection of cell surfaces against complement activation in susceptible individuals. Here, we identified a subgroup of aHUS patients showing persistent activation of the complement alternative pathway and found within this subgroup two families with mutations in the gene encoding factor B (BF), a zymogen that carries the catalytic site of the complement alternative pathway convertase (C3bBb). Functional analyses demonstrated that F286L and K323E aHUS-associated BF mutations are gain-of-function mutations that result in enhanced formation of the C3bBb convertase or increased resistance to inactivation by complement regulators. These data expand our understanding of the genetic factors conferring predisposition to aHUS, demonstrate the critical role of the alternative complement pathway in the pathogenesis of aHUS, and provide support for the use of complement-inhibition therapies to prevent or reduce tissue damage caused by dysregulated complement activation.


Deciphering the transcriptional regulation of the catabolism of lignin-derived aromatics in Rhodococcus opacus PD630.
Rhodococcus opacus PD630 has considerable potential as a platform for valorizing lignin due to its innate "biological funneling" pathways. However, the transcriptional regulation of the aromatic catabolic pathways and the mechanisms controlling aromatic catabolic operons in response to different aromatic mixtures are still underexplored. Here, we identified and studied the transcription factors for aromatic degradation using GFP-based sensors and comprehensive deletion analyses. Our results demonstrate that the funneling pathways for phenol, guaiacol, 4-hydroxybenzoate, and vanillate are controlled by transcriptional activators. The two different branches of the β-ketoadipate pathway, however, are controlled by transcriptional repressors. Additionally, promoter activity assays revealed that the substrate hierarchy in R. opacus may be ascribed to the transcriptional cross-regulation of the individual aromatic funneling pathways. These results provide clues to clarify the molecule-level mechanisms underlying the complex regulation of aromatic catabolism, which facilitates the development of R. opacus as a promising chassis for valorizing lignin.


[Palliative and supportive therapy in cases of renal cell carcinoma].
At the time of diagnosis, 25-30% of all patients with renal cell carcinoma already present with metastatic disease. Furthermore, 20-30% of patients with renal cell carcinoma will have progressive disease despite radical nephrectomy with complete tumor resection. In this review, we discuss the current therapeutic options for patients with metastatic renal cell carcinoma: These include palliative radical nephrectomy, surgery of metastasis, tumor embolisation and medical treatment options (e.g. immunotherapy, chemotherapy and targeted therapy), as well as supportive pain treatment.


Efficacy of Nepafenac versus Flurbiprofen in Maintaining Intraoperative Mydriasis During Phacoemulsification: A Comparative Study.
To compare the efficacy of topical nepafenac (0.1%) with flurbiprofen (0.03%) in maintaining intra-operative mydriasis during phacoemulsification surgery. This study comprised of 160 patients, who were divided into two arms of 80 each (arms A and B) after randomisation. Pre-operatively, all patients received one drop of tropicamide 0.8% and phenylephrine 5% (combination), 4 times, at an interval of 15 minutes on the day of surgery. Thereafter, Nepafenac drop in arm A/Flurbiprofen drop in arm B was administered 4 times, at an interval of 15 minutes keeping a gap of 10 minutes between tropicamide-phenylephrine and any of the experimental drugs. Phacoemulsification was performed one hour after the administration of last drop. Both vertical and horizontal pupillary diameter were measured at three steps; immediately before the surgical incision (baseline), at the end of emulsification of nucleus (before irrigation and aspiration) and at the end of surgery (after stromal hydration). The difference in pupillary diameter between two groups, was statistically insignificant for vertical diameter (P = 0.08) and horizontal diameter (P = 0.28) at the start of surgery. On the other hand, pupillary diameter difference was statistically significant after emulsification of nucleus and at the end of surgery as well when both vertical (P < 0.05) and horizontal diameter (P < 0.05) were considered. The total reduction in pupillary diameter (both vertically and horizontally) was significantly less in the Nepafenac as compared to Flurbiprofen group (P < 0.05). Analysis of mean cumulative dissipated energy did not document any appreciable difference between the two groups. Phacoemulsification time analysis yielded statistically significant results (P = 0.004) between the Nepafenac and Flurbiprofen group. In the present study, topical Nepafenac (0.1%) proved to be more efficacious in maintaining intra-operative mydriasis during phacoemulsification surgery as compared to topical Flurbiprofen (0.03%).


[Seasonal dynamics of the main helminthiases on a sheep farm pasture].
Two-year observations were made on the seasonal and age dynamic of the basic helminth infections on a pasture complex of the village of Bochkovtsi , Veliko Turnovo district. The methods of F ulleborn , Berman , successive washing, and MacMaster were employed to investigate a total of 2198 fecal samples taken from 50 permanently marked lambs, weaned lambs, and ewes. The parasite burden and the species composition of the helminths were established via complete post-mortem examinations, resp., helminthologic necropsies of 19 ewes, 7 weaned lambs, and 5 lambs. Studied were also the surrounding field conditions in order to discover and investigate the biotopes. A total of 517 fresh-water snails of the Limnea truncatula species and 640 land ones of the Helicella obvia species were examined. The helminth fauna was established with the various categories of animals raised on the pasture complex along with the seasonal dynamic of the basic parasitic diseases. A programme for the prophylaxis and therapy was worked out.


Multifunctional roles of the conserved Arg residues in the second region of homology of p97/valosin-containing protein.
The 97-kDa molecular chaperone valosin-containing protein (VCP) belongs to a highly conserved AAA family and forms a hexameric structure that is essential for its biological functions. The AAA domain contains highly conserved motifs, the Walker A, Walker B, and the second region of homology (SRH). Although Walker A and B motifs mediate ATP binding and hydrolysis, respectively, the function of the SRH in VCP is not clear. We examined the significance of the SRH in VCP, especially the conserved Arg(359) and Arg(362) in the first AAA domain, D1 and Arg(635) and Arg(638) in the second AAA domain, D2. We show that Arg(359) and Arg(362) in D1 are critical for maintaining the hexameric structure and the ability to bind the polyubiquitin chains. Although the rest of the tested SRH mutants retain the hexameric structure, all of them exhibit severely reduced ATPase activity. Tryptophan fluorescence analysis showed that all of the tested mutants can bind to ATP or ADP. Thus, the reduced ATPase activity likely results from the hampered communications among protomers during hydrolysis. Moreover, when the ATPase-defective mutant R635A or R638A is mixed with the Walker A mutant of D2, the ATPase activity is partially restored, suggesting that Arg(635) and Arg(638) can stimulate the ATPase activity of the neighboring protomer. Interestingly, mutation of Arg(359) and Arg(362) uncouples the inhibitory effect of p47, a VCP co-factor, on the ATPase activity of VCP. Therefore, the Arg residues allow D1 to take on a specific conformation that is required for substrate binding and co-factor communications. Taken together, these results demonstrate that the conserved Arg residues in the SRH of both D1 and D2 play critical roles in communicating the conformational changes required for ATP hydrolysis, and SRH in D1 also contributes to substrate binding and co-factor communications.


Association study of MAO-A, COMT, 5-HT2A, DRD2, and DRD4 polymorphisms with illness time course in mood disorders.
The aim of our study was to investigate a possible influence of monoamine oxydase A (MAO-A), catechol-O-methyltransferase (COMT), serotonin receptor 2A (5-HT2A), dopamine receptor D2 (DRD2), and dopamine receptor D4 (DRD4) gene variants on timing of recurrence in mood disorders. Gene variants were determined using PCR-based techniques in 550 inpatients affected by recurrent mood disorders (major depressives: n = 212; bipolars: n = 338), rapid cycling mood disorder (n = 81), and 663 controls. We investigated possible genetic influences by comparing illness time course of subjects subdivided according to genotype using multivariate analysis of variance (MANOVA). We could not observe a significantly different time course. No demographic and clinical variables such as sex, age or polarity of onset, presence of psychotic features, genetic loading, or education level influenced the observed results. Our results suggest that MAO-A, COMT, 5-HT2A, DRD2, and DRD4 gene variants are not involved in susceptibility toward different time courses in mood disorders.


Primitive Tumour of the Pulmonary Valve: Discussion of the Differential Diagnosis.
There is a paucity of information concerning cardiac tumours of the pulmonary valve due to their rarity at this location. We report a case of a 47-year-old patient suffering from haemoptysis, asthenia, and acute kidney injury (AKI). A transthoracic echocardiography (TTE) revealed a mass on the pulmonary valve. Further diagnostic investigation was completed until he exhibited worsening hemodynamic instability. This case emphasizes the lack of information regarding the management of a pulmonary valve tumour.


Ultrastructure of neutrophilic phagosome of autologous platelet in vivo in specific granule deficiency.
The ultrastructure of neutrophilic phagosomes containing autologous platelets in vivo from a woman with specific granule deficiency is described. This phagosome formation was observed only when she had developed pneumonia due to Staphylococcus aureus. Therefore, the phenomenon might be induced by severe bacterial infection under an abnormal neutrophilic condition accompanied by specific granule deficiency.


Differential roles of vascular endothelial growth factor receptor-1 and receptor-2 in angiogenesis.
Vascular endothelial growth factor (VEGF)-A, a major regulator for angiogenesis, binds and activates two tyrosine kinase receptors, VEGFR1 (Flt-1) and VEGFR2 (KDR/Flk-1). These receptors regulate physiological as well as pathological angiogenesis. VEGFR2 has strong tyrosine kinase activity, and transduces the major signals for angiogenesis. However, unlike other representative tyrosine kinase receptors which use the Ras pathway, VEGFR2 mostly uses the Phospholipase-Cgamma-Protein kinase-C pathway to activate MAP-kinase and DNA synthesis. VEGFR2 is a direct signal transducer for pathological angiogenesis including cancer and diabetic retinopathy, thus, VEGFR2 itself and the signaling appear to be critical targets for the suppression of these diseases. VEGFR1 plays dual role, a negative role in angiogenesis in the embryo most likely by trapping VEGF-A, and a positive role in adulthood in a tyrosine kinase-dependent manner. VEGFR1 is expressed not only in endothelial cells but also in macrophage-lineage cells, and promotes tumor growth, metastasis, and inflammation. Furthermore, a soluble form of VEGFR1 was found to be present at abnormally high levels in the serum of preeclampsia patients, and induces proteinurea and renal dysfunction. Therefore, VEGFR1 is also an important target in the treatment of human diseases. Recently, the VEGFR2-specific ligand VEGF-E (Orf-VEGF) was extensively characterized. Interestingly, the activation of VEGFR2 via VEGF-E in vivo results in a strong angiogenic response in mice with minor side effects such as inflammation compared with VEGF-A, suggesting VEGF-E to be a novel material for pro-angiogenic therapy.


The genotoxic effects of DNA lesions induced by artificial UV-radiation and sunlight.
Solar radiation sustains and affects all life forms on Earth. The increase in solar UV-radiation at environmental levels, due to depletion of the stratospheric ozone layer, highlights serious issues of social concern. This becomes still more dramatic in tropical and subtropical regions where radiation-intensity is still higher. Thus, there is the need to evaluate the harmful effects of solar UV-radiation on the DNA molecule as a basis for assessing the risks involved for human health, biological productivity and ecosystems. In order to evaluate the profile of DNA damage induced by this form of radiation and its genotoxic effects, plasmid DNA samples were exposed to artificial-UV lamps and directly to sunlight. The induction of cyclobutane pyrimidine dimer photoproducts (CPDs) and oxidative DNA damage in these molecules were evaluated by means of specific DNA repair enzymes. On the other hand, the biological effects of such lesions were determined through the analysis of the DNA inactivation rate and mutation frequency, after replication of the damaged pCMUT vector in an Escherichia coliMBL50 strain. The results indicated the induction of a significant number of CPDs after exposure to increasing doses of UVC, UVB, UVA radiation and sunlight. Interestingly, these photoproducts are those lesions that better correlate with plasmid inactivation as well as mutagenesis, and the oxidative DNA damages induced present very low correlation with these effects. The results indicated that DNA photoproducts play the main role in the induction of genotoxic effects by artificial UV-radiation sources and sunlight.


USP21 modulates Goosecoid function through deubiquitination.
The homeobox gene Goosecoid (GSC), which is known to regulate craniofacial development, is activated by mono-ubiquitination; however, the deubiquitylase responsible for GSC deubiquitination and inhibition has yet to be identified. In the present study, we constructed the recombinant plasmid pFlag-CMV-2-GSC and the SRY (sex-determining region Y)-box 6 (Sox6) reporter gene system to identify deubiquitylases that regulate GSC expression. We demonstrate that the ubiquitin carboxyl-terminal hydrolase 21 (USP21) regulates the deubiquitination of GSC negatively, as demonstrated by its inhibition of Sox6 reporter gene transcription. USP21 interacted with GSC to promote GSC deubiquitination while having no effect on GSC protein stability. Cell viability, migration, and function in ATDC5 cells were probably influenced by USP21 through GSC. These findings suggest that USP21 modulates GSC function through deubiquitination.


Management Strategy of Alzheimer's Patients under the Medical-Care Integration Model Based on Big Data Evaluation.
In the context of the era of big data, the management of Alzheimer's patients has aroused widespread concern in the society. What should the elderly and people with dementia do and how the society should accommodate these special groups have aroused heated discussions in the society. The goal of the medical-nursing integration model is to realize the integrated model of medical care-nursing-rehabilitation-old care, and to better satisfy the needs by providing medical services, rehabilitation care, and health management. The medical needs of the elderly can help the elderly maintain a healthy state; the elderly care needs can be better met through elderly care services, and the burden on families and society can be reduced. With the advancement of the medical-care integration model, new solutions have been provided for the management of senile dementia patients. Therefore, under the medical-care integration model, this paper managed Alzheimer's patients based on the big data algorithm based on association rules and compared it with the management of Alzheimer's patients under the traditional model, and drew the following conclusions: compared with the management satisfaction of Alzheimer's patients under the traditional model, family members and patients' total satisfaction with nursing management under the medical-care integration model was greatly improved; compared with the management of Alzheimer's patients under the traditional model, the incidence of adverse events in the management of Alzheimer's patients under the medical-care integration model was greatly reduced, which was reduced to 17.6%; under the medical-care integration model, the anxiety and depression scores of the elderly patients with dementia decreased rapidly; under the medical-care integration model, the living ability of senile dementia patients has been greatly improved; the use of the medical-care integration model to manage senile dementia patients has greatly improved the sleep quality, mental quality, and quality of life of senile dementia patients.


Effects of dietary β-mannanase supplementation on the additivity of true metabolizable energy values for broiler diets.
This experiment was conducted to determine the effects of dietary β-mannanase on the additivity of true metabolizable energy (TME) and nitrogen-corrected true metabolizable energy (TMEn) for broiler diets. A total of 144 21-day-old broilers were randomly allotted to 12 dietary treatments with 6 replicates. Five treatments consisted of 5 ingredients of corn, wheat, soybean meal, corn distillers dried grains with solubles, or corn gluten meal. One mixed diet containing 200 g/kg of those 5 ingredients also was prepared. Additional 6 treatments were prepared by mixing 0.5 g/kg dietary β-mannanase with those 5 ingredients and the mixed diet. Based on a precision-fed chicken assay, TME and TMEn values for 5 ingredients and the mixed diet as affected by dietary β-mannanase were determined. Results indicated that when β-mannanase was not added to the diet, measured TME and TMEn values for the diet did not differ from the predicted values for the diet, which validated the additivity. However, for the diet containing β-mannanase, measured TMEn value was greater (p<0.05) than predicted TMEn value, indicating that the additivity was not validated. In conclusion, the additivity of energy values for the mixed diet may not be guaranteed if the diet contains β-mannanase.


An examination of posttraumatic, postsurgical orbital deformities: conclusions drawn for improvement of primary treatment.
A retrospective analysis was carried out on the records of 317 patients operated on by the senior author (S.A.W.) for orbital fractures between 1975 and 2007. Two hundred forty of the patients had been previously operated on elsewhere and required further correction (posttraumatic, postsurgical orbital deformity). A smaller group of patients (n = 77) were operated on primarily. The two groups were not, of course, similar, because the posttraumatic, postsurgical orbital deformity group had been operated on by a variety of surgeons with varying levels of experience and ability, and the group of patients operated on primarily had a larger percentage of fractures in the pediatric age group, because of the practice being partially based in a children's hospital, and a larger percentage of severe, compound orbital injuries, because of statewide referrals. Nevertheless, a number of causes for reoperation seen in the posttraumatic, postsurgical orbital deformity group were not seen in the primarily operated group. These included lower eyelid retraction attributable to use of the subciliary incision, displacement and extrusion of alloplastic materials, and fixation of fractures in a nonreduced position. These differences validate, in the authors' opinion, the application of the basic principles of craniofacial reconstruction set forth by Paul Tessier, listed in the text, to both the primary and secondary treatment of posttraumatic orbital deformities.


Thiazoles and Thiazolidinones as COX/LOX Inhibitors.
Inflammation is a natural process that is connected to various conditions and disorders such as arthritis, psoriasis, cancer, infections, asthma, etc. Based on the fact that cyclooxygenase isoenzymes (COX-1, COX-2) are responsible for the production of prostaglandins that play an important role in inflammation, traditional treatment approaches include administration of non-steroidal anti-inflammatory drugs (NSAIDs), which act as selective or non-selective COX inhibitors. Almost all of them present a number of unwanted, often serious, side effects as a consequence of interference with the arachidonic acid cascade. In search for new drugs to avoid side effects, while maintaining high potency over inflammation, scientists turned their interest to the synthesis of dual COX/LOX inhibitors, which could provide numerous therapeutic advantages in terms of anti-inflammatory activity, improved gastric protection and safer cardiovascular profile compared to conventional NSAIDs. Τhiazole and thiazolidinone moieties can be found in numerous biologically active compounds of natural origin, as well as synthetic molecules that possess a wide range of pharmacological activities. This review focuses on the biological activity of several thiazole and thiazolidinone derivatives as COX-1/COX-2 and LOX inhibitors.


Pressure dependence of amide hydrogen-deuterium exchange rates for individual sites in T4 lysozyme.
We report measurements of the pressure dependence of rate constants for the exchange of amide residue protons with solvent deuterium for T4 lysozyme. Data obtained at nine pressures from 0.1 to 200 MPa are analyzed using an elementary kinetic model and the formalism of transition state theory which yield activation volumes for the exchange process. Resolution of individual amide sites was accomplished using the HSQC two-dimensional (2D) NMR experiment on uniformly (15)N-labeled protein. The observed activation volumes span the range from 2.75 to -25.1 mL/mol at 22 degreesC and pH* 7.5. When corrected for the pressure dependence of the ionic product for water and for the reported activation volume for the amide exchange reaction in model compounds, the portion of the activation volume associated with the accessibility of the solvent or catalyst to the amide sites ranges from -15.1 to 12.8 mL/mol. There is no simple correlation between the activation volumes and the protection factors for amide hydrogen exchange. The activation volumes for residues in close proximity in either the primary sequence or the folded structure may differ considerably. There is no trivial correlation between the activation volume and the secondary structural unit in which a residue is located, and activation volumes for residues that are apparently structurally coupled may be very different. The modest sizes of the activation volumes obtained under these conditions are in contrast to large values reported for bovine pancreatic trypsin inhibitor at more extreme conditions of 60 degreesC and pH* 8 where major unfolding events or structural rearrangements may dominate the mechanism [Wagner, G. (1983) Q. Rev. Biophys. 16, 1-57].


A subpopulation of efferent neurons in the mouse suprachiasmatic nucleus is also light responsive.
The suprachiasmatic nucleus (SCN) is the site of a circadian clock with input (afferent) pathways for photic entrainment and output (efferent) pathways for expression of overt, measurable rhythms. To determine whether there are individual neurons in the mouse SCN that might be part of both pathways, we performed double-label immunohistochemistry for light-induced c-Fos and the retrograde tracer cholera toxin subunit B (CtB), 2 weeks after CtB was iontophoresed into the subparaventricular area (subPVA). A minority of neurons was found that were both efferent to the subPVA and responsive to light. This cellular subset may function as a direct channel through the SCN for photic inputs to influence neural outputs, and its existence highlights the topographical heterogeneity of SCN tissue.


Carbonic anhydrase activation enhances object recognition memory in mice through phosphorylation of the extracellular signal-regulated kinase in the cortex and the hippocampus.
Rats injected with by d-phenylalanine, a carbonic anhydrase (CA) activator, enhanced spatial learning, whereas rats given acetazolamide, a CA inhibitor, exhibited impairments of fear memory consolidation. However, the related mechanisms are unclear. We investigated if CAs are involved in a non-spatial recognition memory task assessed using the object recognition test (ORT). Systemic administration of acetazolamide to male CD1 mice caused amnesia in the ORT and reduced CA activity in brain homogenates, while treatment with d-phenylalanine enhanced memory and increased CA activity. We provided also the first evidence that d-phenylalanine administration rapidly activated extracellular signal-regulated kinase (ERK) pathways, a critical step for memory formation, in the cortex and the hippocampus, two brain areas involved in memory processing. Effects elicited by d-phenylalanine were completely blunted by co-administration of acetazolamide, but not of 1-N-(4-sulfamoylphenyl-ethyl)-2,4,6-trimethylpyridinium perchlorate (C18), a CA inhibitor that, differently from acetazolamide, does not cross the blood brain barrier. Our results strongly suggest that brain but not peripheral CAs activation potentiates memory as a result of ERK pathway enhanced activation.


Origins of the Stereoselectivity in a Thiourea-Primary Amine-Catalyzed Nazarov Cyclization.
The origins of stereoselectivity of the Nazarov reactions of α-hydroxydivinylketones catalyzed by a vicinal thiourea-primary amine first reported by Tius have been explored with density functional theory. The electrocyclization transition structures in which the thiourea group of the catalyst donates two hydrogen bonds to the keto carbonyl group of the Nazarov reactant and the primary amine accepts a hydrogen bond from the hydroxyl group of the reactant have been modeled. The enantiomeric Nazarov transition structures, which are conventionally described by the absolute sense of conrotation of the dienone termini ("clockwise" or "counterclockwise") in the literature, are nonplanar and adopt helically chiral conformations. The interactions of these helical electrocyclization transition structures with the chiral catalyst are studied in detail. The organocatalyst is found to employ a combination of hydrogen bonding and steric effects to achieve helical recognition of the Nazarov transition state.


Cellular and humoral immunity in Hodgkin's disease. I Patients in continuous long-term remission.
Cell-mediated and humoral immunity to herpes simplex virus (HSV), cytomegalovirus (CMV) and varicella zoster virus (VZV) were examined in 37 patients with Hodgkin's disease in continuous long term remission. This group had lower blast-transformation than a matched control group to all 3 antigens. Patients originally showing B-symptoms had higher transformation to VZV than those with A-symptoms. Patients treated with irradiation only had higher transformation than those treated with either chemotherapy or a combination of chemotherapy and irradiation. There was a clear tendency towards lower transformation in patients having been in remission for 2 years or less. Phytohaemagglutinin (PHA) stimulation gave lower response in the patient group than in the control group. Patients with B-symptoms had lower response than those with A-symptoms. Interferon production was specially impaired in patients with B-symptoms. The patient group had higher CF titers against HSV and CMV while the control group had higher titers against VZV. B-symptom patients had higher titers against VZV than A-symptom patients. It is concluded that HD patients have impaired immune function many years after discontinuation of therapy, but there are certain differences regarding the in vitro immunity within the patient groups.


Atomic force microscopy of height fluctuations of fibroblast cells.
We investigated the nanometer scale height fluctuations of 3T3 fibroblast cells with the atomic force microscope under physiological conditions. A correlation between these fluctuations and lateral cellular motility can be observed. Fluctuations measured on leading edges appear to be predominantly related to actin polymerization-depolymerization processes. We found fast (5 Hz) pulsatory behavior with 1-2 nm amplitude on a cell with low motility showing emphasized structure of stress fibers. Myosin driven contractions of stress fibers are thought to induce this pulsation.


A hominid from the lower Pleistocene of Atapuerca, Spain: possible ancestor to Neandertals and modern humans.
Human fossil remains recovered from the TD6 level (Aurora stratum) of the lower Pleistocene cave site of Gran Dolina, Sierra de Atapuerca, Spain, exhibit a unique combination of cranial, mandibular, and dental traits and are suggested as a new species of Homo-H. antecessor sp. nov. The fully modern midfacial morphology of the fossils antedates other evidence of this feature by about 650, 000 years. The midfacial and subnasal morphology of modern humans may be a retention of a juvenile pattern that was not yet present in H. ergaster. Homo antecessor may represent the last common ancestor for Neandertals and modern humans.


Construction of hybrid regulated mother-specific yeast promoters for inducible differential gene expression.
Engineered promoters with predefined regulation are a key tool for synthetic biology that enable expression on demand and provide the logic for genetic circuits. To expand the availability of synthetic biology tools for S. cerevisiae yeast, we here used hybrid promoter engineering to construct tightly-controlled, externally-inducible promoters that only express in haploid mother cells that have contributed a daughter cell to the population. This is achieved by combining elements from the native HO promoter and from a TetR-repressible synthetic promoter, with the performance of these promoters characterized by both flow cytometry and microfluidics-based fluorescence microscopy. These new engineered promoters are provided as an enabling tool for future synthetic biology applications that seek to exploit differentiation within a yeast population.


Effect of destruction of noradrenergic neurones with DSP4 on performance on a free-operant timing schedule.
This experiment examined the effect of destroying central noradrenergic neurones, using the selective neurotoxin DSP4 [N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine], on performance in a free-operant timing schedule. Rats received either systemic treatment with DSP4 or vehicle-alone injections. They were trained to press levers for a sucrose reinforcer. Training sessions consisted of 40, 50-s trials in which reinforcers were available on a variable-interval 25-s schedule; in the first 25 s of each trial, reinforcers were only available for responses on lever A, whereas in the last 25 s reinforcers were available only for responses on lever B. Data were collected from probe trials (four per session), in which no reinforcers were delivered, during the last ten of 60 training sessions. Both groups showed decreasing response rates on lever A, and increasing response rates on lever B, as a function of time from the onset of the trial. Quantitative indices of timing behaviour were derived from a two-parameter logistic function fitted to the relative response rates on lever B (response rate on lever B, expressed as a percentage of overall response rate); this function accounted for > 90% of the data variance in each group. The DSP4-treated group showed a significantly lower value of the indifference point (i.e. the time corresponding to 50% responding on lever B) than the control group. The slope of the function and the rate of switching between response alternatives did not differ significantly between the two groups. The concentrations of noradrenaline were markedly reduced in the neocortex and hippocampus of the DSP4-treated group, but the concentrations of dopamine, 5-hydroxytryptamine and 5-hydroxyindoleacetic acid were not significantly altered. It is suggested that results may be consistent with a role of the dorsal ascending noradrenergic pathway in behavioural "arousal".


Laparoscopic splenectomy for sclerosing angiomatoid nodular transformation of the spleen.
Sclerosing angiomatoid nodular transformation (SANT) is a rare, benign, proliferative vascular lesion that arises from the splenic red pulp. Most patients with SANT have no clinical symptoms and are discovered incidentally on imaging. There are no definitive radiological signs and a distinction from other splenic diseases, and malignant processes remain difficult. Confirmation of the diagnosis of SANT requires a histological and immunohistochemical evaluation of the resected spleen. Here, we report an unusual case of SANT of the spleen successfully treated with an elective laparoscopic splenectomy (LS). LS is a safe and effective method for diagnosis of SANT.


Effusion under the microscope.
Pseudochylothorax is a very rare form of pleural effusion. It is also called chyliform or cholesterol pleural effusion. It is usually a unilateral process and approximately one-third of patients are asymptomatic at presentation. We report a case of a 60 year old man with a background of rheumatoid arthritis who presented with progressive dyspnea. Chest X-ray revealed a new left pleural effusion and a small persistent right pleural effusion. He presented 5 years prior due to recurrent pleural effusion and no diagnosis was made. Repeat thoracentesis yielded 350 milliliters of thick, milky, tan-colored fluid.


In vitro selection and characterization of cellulose-binding DNA aptamers.
Many nucleic acid enzymes and aptamers have modular architectures that allow them to retain their functions when combined with other nucleotide sequences. This modular function facilitates the engineering of RNAs and DNAs that have more complex functions. We sought to create new DNA aptamers that bind cellulose to provide a module for immobilizing DNAs. Cellulose has been used in a variety of applications ranging from coatings and films to pharmaceutical preparations, and therefore DNA aptamers that bind cellulose might enable new applications. We used in vitro selection to isolate aptamers from a pool of random-sequence DNAs and subjected two distinct clones to additional rounds of mutagenesis and selection. One aptamer (CELAPT 14) was chosen for sequence minimization and more detailed biochemical analysis. CELAPT 14 aptamer variants exhibit robust binding both to cellulose powder and paper. Also, an allosteric aptamer construct was engineered that exhibits ATP-mediated cellulose binding during paper chromatography.


Model of Disc Degeneration in Rat Tail Induced Through a Vascular Isolation of Vertebral Endplates.
Back pain is a major health problem. The degenerative cascade of the spine begins in the intervertebral disc, due to an impairment in the blood supply through the vertebral endplates. Our objective was to develop a novel disc degeneration model based on these premises, akin to the process in humans, in contrast to other proposed models (puncture, enzyme injection, aberrant loads,…) Material and methods: 37 Sprague-Dawley rats, 2 arms: (a) histological (n = 17, one died), en- bloc sections, Van Gieson staining, (Nisimura-Mochida criteria) and also collagen VI staining (tissue oxidative stress), four animals were euthanized every 2 weeks (2-8); and (b) imaging (n = 20, six wound sloughs), MRI 9.4 Tesla protocol, sequential disc volumetric analysis (24 h-8 weeks) in all animals. Disc degeneration was induced by means of vascular isolation of tail discs endplates either from one side or both. Isolation from both sides caused a progressive degeneration of the disc (p < 0.001 vs. controls), bigger than isolation from one side (p < 0.01 vs. both sides and p < 0.05 vs. controls), as rated by volumetric reduction; furthermore, tissue structural changes (Nisimura-Mochida) and collagen VI deposition confirmed these results. the model here described represents a novel and translational tool that reproduces the intervertebral disc degeneration in a similar way to that taking place in human beings.


Eligibility for alirocumab or evolocumab treatment in 1090 hypercholesterolemic patients referred to a regional cholesterol treatment center with LDL cholesterol ≥70 mg/dL despite maximal-tolerated LDL-cholesterol-lowering therapy.
Proprotein convertase subtilisin/kexin type 9 inhibitors, Praluent (alirocumab [ALI]) and Repatha (evolocumab [EVO]) have been approved as adjuncts to the standard-of-care maximal-tolerated dose (MTD) of low-density lipoprotein cholesterol (LDLC)-lowering therapy (LLT), statin therapy, in heterozygous (HeFH) (ALI or EVO) or homozygous (EVO) familial hypercholesterolemia, or clinical atherosclerotic cardiovascular disease (CVD) where LDLC lowering is insufficient (both). Since LDLC lowering has been revolutionized by ALI and EVO, specialty pharmaceutical pricing models will be applied to a mass market. We applied US Food and Drug Administration (FDA) and insurance eligibility criteria for ALI and EVO to 1090 hypercholesterolemic patients serially referred over 3 years who then received ≥2 months maximal-tolerated dose of standard-of-care LDL cholesterol-lowering therapy (MTDLLT) with follow-up LDLC ≥70 mg/dL. MTDLLT did not include ALI or EVO, which had not been commercially approved before completion of this study. Of the 1090 patients, 140 (13%) had HeFH by clinical diagnostic criteria and/or CVD with LDLC >100 mg/dL despite ≥2 months on MTDLLT, meeting FDA insurance criteria for ALI or EVO therapy. Another 51 (5%) patients were statin intolerant, without HeFH or CVD. If 13% of patients with HeFH-CVD and LDLC >100 mg/dL despite MTDLLT are eligible for ALI or EVO, then specialty pharmaceutical pricing models (~$14,300/year) might be used in an estimated 10 million HeFH-CVD patients. Whether the health care savings arising from the anticipated reduction of CVD events by ALI or EVO justify their costs in populations with HeFH-CVD and LDLC >100 mg/dL despite MTDLLT remains to be determined.


Kinetics of DNA hydration.
The hydration of the d(CGCGAATTCGCG) B-DNA duplex in solution was studied by nuclear magnetic relaxation dispersion (NMRD) of the water nuclei 1H, 2H, and 17O, and by nuclear Overhauser effects (NOEs) in high-resolution two-dimensional 1H NMR spectra. By comparing results from the free duplex with those from its complex with netropsin, water molecules in the "spine of hydration" in the AATT region of the minor groove could be distinguished from hydration water elsewhere in the duplex. The 2H and 17O relaxation dispersions yield a model-independent residence time of 0.9(+/-0.1) ns at 4 degrees C for five highly ordered water molecules in the spine. When corrected for frequency offset effects, the NOE data yield the same residence time as the NMRD data, giving credence to both methods. At 27 degrees C, the residence time is estimated to 0.2 ns, a factor of 40 shorter than the tumbling time of the duplex. The NMRD data show that all water molecules associated with the duplex, except the five molecules in the spine, have residence times significantly shorter than 1 ns at 4 degrees C. There is thus no long-lived hydration structure associated with the phosphate backbone. In contrast to 2H and 17O, the 1H relaxation dispersion is dominated by labile DNA protons and therefore provides little information about DNA hydration.


Amino acid and lactate catabolism in trimethylamine oxide respiration of Alteromonas putrefaciens NCMB 1735.
The nonfermentative Alteromonas putrefaciens NCMB 1735 grew anaerobically in defined media with trimethylamine oxide as external electron acceptor. All amino acids tested, except taurine and those with a cyclic or aromatic side chain, were utilized during trimethylamine oxide-dependent anaerobic growth. Lactate, serine, and cysteine (which are easily converted to pyruvate) and glutamate and aspartate (which are easily converted to tricarboxylic acid cycle intermediates) were metabolized at the fastest rate. Growth with lactate as growth-limiting substrate gave rise to the formation of 40 mol% acetate, whereas serine and cysteine were nearly completely oxidized to CO2. Molar growth yields with the latter substrates were the same and were 50% higher than with lactate. This showed that more ATP was formed when acetyl coenzyme A entered the tricarboxylic acid cycle than when it was converted via acetyl phosphate to acetate. Also, growth with formate as substrate indicated that the reduction of trimethylamine oxide to trimethylamine was coupled with energy conservation by a respiratory mechanism.


Application of Mosher's method for absolute configuration assignment to bioactive plants and fungi metabolites.
The review deals with the application of the NMR advanced Mosher's method for the assignment of the absolute configuration to plant and fungal metabolites belonging to different classes of natural compounds. The structural and stereochemical characterization of these naturally occurring metabolites, as well as their biological properties as possible drugs or agrochemicals is reviewed. The importance of the absolute and relative stereochemistry on their biological properties is also highlighted. Successes and failures of application of the advanced Mosher's method are reported.


Influence of coronary heart disease on morbidity and mortality after lower extremity revascularization surgery: a population-based study in Olmsted County, Minnesota (1970-1987).
The aim of this study was to evaluate the short- and long-term postoperative cardiac outcome of patients undergoing lower extremity revascularization surgery in a geographically defined patient group. Among patients with peripheral vascular disease, cardiac events have an important effect on long-term outcome after peripheral vascular surgery. However, long-term outcome is poorly documented. We examined the entire community medical records of 173 residents of Olmsted County, Minnesota, who underwent peripheral artery bypass surgery between 1970 and 1987 and were followed up to January 1, 1991. Patients were allocated to subgroups of 60 patients with and 106 patients without overt coronary artery disease. There were no significant differences in perioperative death, myocardial infarction or stroke between subgroups at 30 days after operation. The 5- and 10-year Kaplan-Meier survival rate after operation was 77% and 51% in those without and 54% and 24% in those with overt coronary artery disease (p < 0.001), respectively. For both groups, survival was significantly poorer than that expected for an age- and gender-matched group. Patients undergoing aortoiliac surgery were more likely to be alive at 10 years than those undergoing femoropopliteal surgery (47% vs. 28%, p = 0.001). The 5-year cumulative incidence of cardiac events was greater in those with overt coronary artery disease (50% vs. 28%, p = 0.003). In multivariable analysis, age, coronary artery disease and diabetes were independent predictors of death. Coronary events are the most important cause of long-term morbidity and mortality after peripheral vascular surgery. Patients without overt coronary artery disease are at significant risk for long-term cardiac events.


Brolucizumab 12- and 16-Week Fixed Dosing Potential in Neovascular AMD: A post hoc Evaluation of Data from the HAWK and HARRIER Trials.
This post hoc analysis applies a fixed dosing stratification approach to patient-level brolucizumab data from the phase III HAWK and HARRIER trials to determine the proportion of patients who would have been assigned to fixed dosing regimens with treatment intervals of 8, 12, or 16 weeks (q8w, q12w, or q16w) based on the presence/absence of disease activity (DA) following the loading phase. The analysis also simulates central subfield thickness (CSFT) data to estimate the anatomical outcomes if the patients had been thus assigned. Of note, the limitations of this analysis include the post hoc nature of the work and the inability to directly compare HAWK and HARRIER with TENAYA and LUCERNE due to the differences in design. This study was a post hoc modelling analysis of patient-level data. Using patient-level data from HAWK and HARRIER, patients (n = 730) were allocated to a fixed q16w, q12w, or q8w regimen based on assessment of DA at weeks 16 and 20. Two definitions of DA were used: DA 1, based on a phase II study of faricimab, and DA 2, a definition derived from common clinical consideration including visual acuity and anatomical changes. CSFT simulations were performed using a pharmacokinetic/pharmacodynamic model describing CSFT response to anti-VEGF treatment. Outcome measures were modelled patient allocation to fixed regimens and mean CSFT reduction. Using DA definitions 1 and 2, respectively, 78% and 76% of patients in the brolucizumab arm were allocated to a greater than or equal to q12w regimen, and 56% and 52% were allocated to a q16w regimen. Mean reduction in CSFT was similar between the two study drugs with both DA definition assumptions. This analysis demonstrates the potential durability of action and effectiveness of brolucizumab.


Transforming Growth Factor-β-Activated Kinase 1 Is Required for Human FcγRIIIb-Induced Neutrophil Extracellular Trap Formation.
Neutrophils (PMNs) are the most abundant leukocytes in the blood. PMN migrates from the circulation to sites of infection where they are responsible for antimicrobial functions. PMN uses phagocytosis, degranulation, and formation of neutrophil extracellular traps (NETs) to kill microbes. Several stimuli, including bacteria, fungi, and parasites, and some pharmacological compounds, such as Phorbol 12-myristate 13-acetate (PMA), are efficient inducers of NETs. Antigen-antibody complexes are also capable of inducing NET formation. Recently, it was reported that FcγRIIIb cross-linking induced NET formation similarly to PMA stimulation. Direct cross-linking of FcγRIIA or integrins did not promote NET formation. FcγRIIIb-induced NET formation presented different kinetics from PMA-induced NET formation, suggesting differences in signaling. Because FcγRIIIb also induces a strong activation of extracellular signal-regulated kinase (ERK) and nuclear factor Elk-1, and the transforming growth factor-β-activated kinase 1 (TAK1) has recently been implicated in ERK signaling, in the present report, we explored the role of TAK1 in the signaling pathway activated by FcγRIIIb leading to NET formation. FcγRIIIb was stimulated by specific monoclonal antibodies, and NET formation was evaluated in the presence or absence of pharmacological inhibitors. The antibiotic LL Z1640-2, a selective inhibitor of TAK1 prevented FcγRIIIb-induced, but not PMA-induced NET formation. Both PMA and FcγRIIIb cross-linking induced phosphorylation of ERK. But, LL Z1640-2 only inhibited the FcγRIIIb-mediated activation of ERK. Also, only FcγRIIIb, similarly to transforming growth factor-β-induced TAK1 phosphorylation. A MEK (ERK kinase)-specific inhibitor was able to prevent ERK phosphorylation induced by both PMA and FcγRIIIb. These data show for the first time that FcγRIIIb cross-linking activates TAK1, and that this kinase is required for triggering the MEK/ERK signaling pathway to NETosis.


Real-time clinical alerting: effect of an automated paging system on response time to critical laboratory values--a randomised controlled trial.
Timely and reliable communication of critical laboratory values is a Joint Commission National Patient Safety Goal. The objective was to evaluate the effect of an automated system for paging critical values directly to the responsible physician. A randomised controlled trial on the general medicine clinical teaching units at an urban academic hospital was conducted from February to May 2006; the unit of randomisation was the critical laboratory value. The intervention was an automated paging system that sent the critical value directly to the responsible physician's pager. The control arm was usual care, which was a telephone call to the patient's ward by the laboratory technician. The primary outcome was response time, defined as the interval between acceptance of the critical value into the laboratory information system to the writing of an order on the patient's chart in response to the critical value. If the time of order was not documented, the time of administration of treatment was used to calculate response time. For primary analysis, 165 critical values were evaluated on 108 patients with full response time data. The median response time was 16 min (IQR 2-141) for the automated paging group and 39.5 min (IQR 7-104.5) for the usual care group (p=0.33). The automated paging system reduced the length of time physicians took to respond to critical laboratory values, but this difference was not statistically significant. Future reseach should evaluate the effects of alerts for conditions that currently do not generate a phone call and the addition of real-time decision support to the critical value alerts.


Omega-3 Eicosapentaenoic Acid (EPA) Rich Extract from the Microalga Nannochloropsis Decreases Cholesterol in Healthy Individuals: A Double-Blind, Randomized, Placebo-Controlled, Three-Month Supplementation Study.
The aim of this trial is to assess the effect of Almega®PL on improving the Omega-3 Index, cardio-metabolic parameters, and other biomarkers in generally healthy individuals. The benefits of long-chain omega-3 fatty acids for cardiovascular health are primarily built upon mixtures of docosahexaenoic (DHA) and eicosapentaenoic acids (EPA). Highly purified EPA therapy has proven to be particularly effective in the treatment of cardiovascular disease, but less is known about the benefits of EPA-only supplementation for the general healthy population. Almega®PL is a polar rich oil (>15%) derived from the microalga Nannochloropsis that contains EPA (>25%) with no DHA. Participants (n = 120) were given a capsule of 1 g/day of either Almega®PL or placebo for 12 weeks. Differences in the Omega-3 Index, cardiometabolic markers, and other general health indicators were measured at the baseline, six, and 12 weeks. Compared to the placebo group, Almega®PL supplementation significantly increased the Omega-3 Index and EPA concentration from 4.96 ± 0.90 and 0.82 ± 0.37% at the baseline to 5.75 ± 0.90 and 1.27 ± 0.36 at week 12, respectively. Very-low-density lipoprotein cholesterol (VLDL) decreased by 25%, which resulted in a significant decrease in total cholesterol compared to the placebo. Interestingly, the decrease in VLDL was not associated with an increase in LDL, which seems to be a benefit associated with EPA-only based formulations. Collectively, these results show that Almega®PL provides a natural EPA-only option to increase EPA and manage cholesterol levels in the general population.


Disseminated histoplasmosis mimicking laryngeal carcinoma from central India--a case report.
A 58 year old male, chronic smoker, with diabetes mellitus (Type II), residing in Madhya Pradesh presented with dysphagia, hoarseness, odynophagia and significant weight loss. Microlaryngoscopy showed reddish ulcerated area involving valleculae, and pharyngo-epiglottic fold. Histopathological examination showed foamy macrophages containing Histoplasma capsulatum. Patient initially responded to Azoles, relapsed three months after stopping the treatment and presented with oral lesions. Biopsy showed H. capsulatum, histologically as well as on culture. The mycelial phase converted to yeast phase on B.H.I.A. at 37 degrees C confirming the isolate as H. capsulatum. The patient responded well to amphotericin B.


Metacognitive interpersonal group therapy for adolescents with avoidant personality disorder: The case of Sofia.
Early intervention studies for adolescents and early adults are required to explore the acceptability and effectiveness of psychological therapies across the full range of personality disorders (PDs) beyond just borderline PD. The main aim of the current paper was to describe a Metacognitive Interpersonal Therapy group adaptation for adolescents (MIT-GA) presenting with PDs featuring overcontrol and social inhibition, and in particular Avoidant PD characteristics. We report findings from a single case of a female adolescent diagnosed with Avoidant PD providing preliminary data on feasibility and the possible effectiveness of MIT-GA. Evidence of acceptability and meaningful clinical gains are described. Results suggest that MIT-GA could be a powerful treatment option in patients with generalized social avoidance who do not currently receive gold-standard psychotherapeutic treatments. This intervention also appears to be effective and cost-effective in initiating significant clinical changes in profiles of young patients with non-borderline PD symptoms.


CEFALIUM-INDUCED BILATERAL TRANSIENT MYOPIA, RETINAL FOLDS, AND FOCAL CHOROIDAL DELAY.
To describe a case of acute bilateral transient myopia, retinal folds, and island of choroidal delay associated with oral administration of Cefalium, a medication commonly prescribed in Brazil for migraine that combines acetaminophen 500 mg, caffeine 40 mg, dihydroergotamine mesylate 1 mg, and metoclopramide hydrochloride 10 mg. A 21-year-old woman with bilateral blurred vision 1 day after the use of Cefalium. The main outcomes measures were BCVA, ocular fundus, ocular coherence tomography, and angiography findings. The patient developed bilateral myopia, retinal folds, and focus choroidal delay 1 day after the administration of oral cefalium. Ocular fundus examination and ocular coherence tomography revealed retinal folds in the internal surface of the retina. Angiography showed focus areas of hypofluorescence in both eyes. Seven days after Cefalium was suspended, all clinical symptoms had resolved, with full recovery from the abnormal findings on ocular fundus, ocular coherence tomography, and angiography. This is the first report that identified and described bilateral transient myopia, retinal folds, and focus choroidal delay secondary the use of Cefalium.


[Lipid composition of Cunninghamella elegans cultivated on n-alkanes].
The ability to oxidize n-alkanes was studied with various species of fungi belonging to the Cunninghamella genus. These fungi are able to assimilate hydrocarbons and to accumulate up to 1.5 g/litre of biomass. The most active strain was Cunninghamella elegans (-) 1204. The amount of lipids formed, and their composition, depended on the length of the carbon chain of oxidized alkane. The content of fat in the cells increased with the length of the hydrocarbon chain. The following lipid fractions have been detected: phospholipids, monoglycerides, diglycerides, triglycerides, sterols, free fatty acids, sterol esters, and hydrocarbons. The qualitative composition of the fractions depended, to a considerable extent, on the n-alkane utilized. Investigation of the fatty-acid composition of intracellular lipids has shown that fatty acids with an even number of carbon atoms are formed from hydrocarbons with an even number of these atoms, while fatty acids both with an even and odd number of carbon atoms are synthesized from hydrocarbons with an odd number of these atoms. The relative content of the acids with the same number of carbon atoms as that of the alkane being utilized increased with the length of the carbon chain.


Extracellular ATP, a danger signal, is recognized by DORN1 in Arabidopsis.
ATP, the universal energy currency of all organisms, is released into the extracellular matrix and serves as a signal among cells, where it is referred to as an extracellular ATP. Although a signalling role for extracellular ATP has been well studied in mammals over the last 40 years, investigations of such a role in plants are at an early stage. Recently, the first plant receptor for extracellular ATP, DOes not Respond to Nucleotides (DORN1), was identified in Arabidopsis thaliana by mutant screening. DORN1 encodes a legume-type lectin receptor kinase that is structurally distinct from the mammalian extracellular ATP receptors. In the present review, we highlight the genetic and biochemical evidence for the role of DORN1 in extracellular ATP signalling, placing this within the wider context of extracellular ATP signalling during plant stress responses.


Silicon nanowire array overcomes chemotherapeutic resistance by inducing the differentiation of breast cancer stem cells.
Currently, traditional cancer treatment strategies are greatly challenged by the existence of cancer stem cells (CSCs), which are root cause of chemotherapy resistance. Differentiation therapy presents a novel therapeutic strategy for CSC-targeted therapy. However, there are very few studies on the induction of CSCs differentiation so far. Silicon nanowire array (SiNWA) with many unique properties is considered to be an excellent material for various applications ranging from biotechnology to biomedical applications. In this study, we report the SiNWA differentiates MCF-7-derived breast CSCs (BCSCs) into non-CSCs by modulating the morphology of cells. In vitro, the differentiated BCSCs lose the stemness properties and thus become sensitive to chemotherapeutic drugs, eventually leading to the death of BCSCs. Therefore, this work suggests a potential approach for overcoming chemotherapeutic resistance.


Novel molecular diagnostic system of limbal stem cell deficiency based on MUC5AC transcript detection in corneal epithelium by PCR-reverse dot blot.
To evaluate the sensitivity and specificity of a PCR-strip system based on reverse dot blot for detection of MUC5AC mRNA in corneal epithelium samples from patients with limbal stem cell deficiency (LSCD), and to determine the correlation with clinical diagnosis. We obtained 87 corneal impression cytology (IC) samples from 55 subjects (37 patients clinically diagnosed with LSCD and 18 control subjects). Total RNA was extracted from each IC sample and retrotranscribed to cDNA. MUC5AC mRNA transcript was amplified by a customized RT-PCR assay and detected in PCR strips based on reverse dot blot hybridization and in agarose gels. Conjunctival IC samples were used as positive controls. Forty-four of 45 corneal IC samples obtained from patients clinically diagnosed with LSCD were positive for MUC5AC, whereas 34 of 42 corneal ICs from healthy subjects were negative for MUC5AC. Four healthy corneas were found MUC5AC positive, and four rendered inconclusive results. A correlation of 91.4% (P < 0.001) between molecular testing and clinical diagnosis was found. The overall sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the PCR-strip system were 98%, 89%, 92%, and 97%, respectively. Corneal epithelium MUC5AC transcript detection by reverse dot blot PCR-strip is a highly sensitive technique for LSCD diagnosis. The test results strongly correlate with clinical diagnosis of characterized LSCD cases. The PCR-strip system may be used for early detection, and for the detection of mild cases of LSCD, and constitutes an objective clinical tool for the monitoring of treatments and surgical decisions.


Molecular basis of the optochin-sensitive phenotype of pneumococcus: characterization of the genes encoding the F0 complex of the Streptococcus pneumoniae and Streptococcus oralis H(+)-ATPases.
The gene responsible for the optochin-sensitive (OptS) phenotype of Streptococcus pneumoniae has been characterized. Sequence comparisons indicated that the genes involved encoded the subunits of the F0 complex of an H(+)-ATPase. Sequence analysis and transformation experiments showed that the atpC gene is responsible for the optochin-sensitive resistant (OptS/OptR) phenotype. Our results also show that natural as well as laboratory OptR isolates have arisen by point mutations that produce different amino acid changes at positions 48, 49 or 50 of the ATPase c subunit. The nucleotide sequence of the F0 complex of the Streptococcus oralis ATPase has also been determined. In addition, comparison of the sequence of the atpCAB genes of S. pneumoniae R6 (OptS) and M222 (an OptR strain produced by interspecies recombination between pneumococcus and S. oralis), and S. oralis revealed that, in M222, an interchange of atpC and atpA had occurred. We also demonstrate that optochin specifically inhibited the membrane-bound ATPase activity of the S. pneumoniae wild-type (OptS) strains, and found a 100-fold difference between OptS and OptR strains, both in growth inhibition and in membrane ATPase resistance.


Effect of Protonation on the Secondary Structure and Orientation of Plant Light-Harvesting Complex II Studied by PM-IRRAS.
The major light-harvesting pigment-protein complex of photosystem II, LHCII, has a crucial role in the distribution of the light energy between the two photosystems, the efficient light capturing and protection of the reaction centers and antennae from overexcitation. In this work direct structural information on the effect of LHCII protonation, which mimics the switch from light-harvesting to photoprotective state of the protein, was revealed by polarization-modulated infrared reflection-absorption spectroscopy (PM-IRRAS). PM-IRRAS on LHCII monolayers verified that the native helical structure of the protein is preserved in both partly deprotonated (pH 7.8, LHCII) and protonated (pH 5.2, p-LHCII) states. At low surface pressure, 10 mN/m, the orientation of the α-helices in these two LHCII states is different-tilted (θ ≈ 40°) in LHCII and nearly vertical (θ ≈ 90°) in p-LHCII monolayers; the partly deprotonated complex is more hydrophilic than the protonated one and exhibits stronger intertrimer interactions. At higher surface pressure, 30 mN/m, which is typical for biological membranes, the protonation affects neither the secondary structure nor the orientation of the transmembrane α-helices (tilted ∼45° relative to the membrane surface in both LHCII states) but weakens the intermolecular interactions within and/or between the trimers.


Functional domains of the RNA component of ribonuclease P revealed by chemical probing of mutant RNAs.
The higher-order structure of the RNA component of ribonuclease P from Escherichia coli was analyzed using chemical probes. The secondary structure model which had been constructed from the comparative sequence analysis of the RNA was refined using the experimental data. In a mutant RNA (A89 RNA), which contains a G----A substitution at nucleotide 89, we detected a number of conformational alterations clustered between nucleotides 90 and 239. In view of the fact that A89 RNA is as catalytically active as wild-type RNA, but defective in association with the protein component, it is clear that the catalytic function of the RNA component resides on the structure which is not disrupted by the A89 mutation and that the structures altered by the mutation represent the region(s) interacting with the protein component. Another mutant (A329 RNA), which has a G----A substitution at nucleotide 329 and is defective in catalytic function, showed no detectable change in higher-order structure.


Role of endogenously secreted angiotensin II in the CO2-induced stimulation of HCO3 reabsorption by renal proximal tubules.
Previous studies demonstrated that the proximal tubule (PT) responds to isolated increases in basolateral ([CO(2)](BL)) or "bath" CO(2) concentration by increasing the HCO(3)(-) reabsorption rate (J(HCO(3))). Blockade of the rabbit apical AT(1) receptor or knockout of the mouse AT(1A) receptor eliminates these effects, demonstrating a requirement for luminal ANG II that the PT itself synthesizes. In the present study, we examined the effects of the ACE inhibitor lisinopril on J(HCO(3)) in isolated perfused rabbit PTs (S2 segment), using out-of-equilibrium solutions to make isolated changes in [CO(2)](BL) at a fixed baseline HCO(3)(-) concentration of 22 mM and fixed baseline pH of 7.4. Adding 60 or 240 nM lisinopril (in vitro K(i): 0.5 or 1.2 nM) to the lumen had no effect. These results are not consistent with the hypothesis that the PT secretes either angiotensinogen or ANG I. However, adding 60 nM basolateral lisinopril significantly decreased J(HCO(3)) at a [CO(2)](BL) of 20%. Moreover, 240 nM basolateral lisinopril decreased baseline (i.e., at 5% CO(2)) J(HCO(3)) by one-half and completely eliminated the response to altering [CO(2)](BL) from 0 to 20%, but left intact the stimulatory effect of 10(-11) M basolateral ANG II. At extremely high concentrations (i.e., 100 microM), luminal lisinopril replicated the effects of 240 nM basolateral lisinopril. Our data are consistent with the hypothesis that lisinopril readily crosses the basolateral (but not apical) membrane to block ACE in a vesicular compartment. We conclude that the isolated PT predominantly secretes preformed ANG II, rather than angiotensinogen or ANG I.


Glucose-regulated protein 78 (GRP78) regulates colon cancer metastasis through EMT biomarkers and the NRF-2/HO-1 pathway.
Glucose-regulated protein 78 (GRP78) is a key chaperone and stress response protein. Previous studies have demonstrated that high GRP78 expression may be correlated with cancer progression and therapeutic response. However, the role of GRP78 in the metastasis of colon cancer is unclear. In this study, we used small interfering RNA (siRNA) to knock down GRP78 expression in colon cancer cells (HT-29 and DLD-1 cells). In wound-healing migration assays, we found that GRP78-knockdown (GRP78KD) cells showed better wound-healing ability than control cells. We also found that GRP78KD cells displayed a better migratory ability than control cells in migration and invasion assays. As we further dissected the underlying molecular mechanism, we found that silencing GRP78 may cause an increase in vimentin expression and a decrease in the E-cadherin level, which was correlated with the increase in migratory ability. In addition, we found that GRP78KD may activate the NRF-2/HO-1 pathway, and this activation was also correlated with the increase in cell invasiveness. Furthermore, we examined GRP78 expression in a tissue array and found that the GRP78 expression in metastatic adenocarcinoma in lymph nodes tended to be weaker than that in primary colonic adenocarcinoma. In conclusion, a low level of GRP78 may cause an increase in metastasis ability in colon cancer cells by altering E-cadherin and vimentin expression and activating the NRF-2/HO-1 signaling pathway. Our study demonstrates that low expression of GRP78 may correlate with a high risk of metastasis in colon cancer.


Separation and size determination of human serum lipoproteins by agarose gel filtration.
A method is described for the separation of the three major classes of human serum lipoproteins by gel filtration on columns of 4 and 6% agarose gel. After calibration of the columns, the elution volumes of the lipoproteins were used to calculate the molecular sizes and molecular weights of these macromolecules. The technique was employed to demonstrate aggregation of low density lipoprotein following partial delipidation, partial proteolysis, or mild heat denaturation. Agarose gel filtration shows promise as a useful method for the isolation, purification, and characterization of lipoproteins.


Resistance to degradation by nucleases of (2'S)-2'-deoxy-2'-C-methyloligonucleotides, novel potential antisense probes.
An oligonucleotide must fulfill two main requirements to become a potential antisense probe: effective hybridization properties with the complementary sequence and stability toward nucleases. In this article the degradation pattern of a new class of potential antisense fragment, (2'S)-2'-deoxy-2'-C-methyloligonucleotides, is analyzed. The results described here show that the modification introduced in these oligonucleotides confers an enhanced stability toward purified nucleases, human sera, and HeLa cell extracts.


Pattern association--a key to recognition of shark attacks.
Investigation of a number of shark attacks in South Australian waters has lead to recognition of pattern similarities on equipment recovered from the scene of such attacks. Six cases are presented in which a common pattern of striations has been noted.


Metabolism and bioactivation of the tricyclic antidepressant amitriptyline in human liver microsomes and human urine.
Amitriptyline is a widely used tricyclic antidepressant, but the metabolic studies were conducted almost 20 years ago using high-performance liquid chromatography coupled with ultraviolet detector or radiolabeled methods. First, multiple ion monitoring (MIM)- enhanced product ion (EPI) scan was used to obtain the diagnostic ions or neutral losses in human liver microsome incubations with amitriptyline. Subsequently, predicted multiple reaction monitoring (MRM)-EPI scan was used to identify the metabolites in human urine with the diagnostic ions or neutral losses. Finally, product ion filtering and neutral loss filtering were used as the data mining tools to screen metabolites. Consequently, a total of 28 metabolites were identified in human urine after an oral administration using LC-MS/MS. An integrated workflow using LC-MS/MS was developed to comprehensively profile the metabolites of amitriptyline in human urine, in which five N-acetyl-l-cysteine conjugates were characterized as tentative biomarkers for idiosyncratic toxicity.


XAF1 expression levels in a non-small cell lung cancer cohort and its potential association with carcinogenesis.
The process of lung carcinogenesis is still not well understood and involves different levels of regulation of several genes. The search for molecular biomarkers, which can be applicable to clinical practice, has been the focus of various studies. XIAP-associated factor 1 (XAF1) was previously shown to be downregulated in many types of tumors, including squamous cell lung cancer. XAF1 is a pro-apoptotic protein and its restoration was found to sensitize cancer cells to apoptotic stimuli; however, the precise mechanism involved in the downregulation of XAF1 in tumors is unknown and promoter hypermethylation or heat-shock transcription factor 1 (HSF1) may be involved. Therefore, the aim of the present study was to evaluate the expression of XAF1 in tumors and adjacent non-tumor specimens from non-small cell lung cancer (NSCLC) patients, and its potential association with various factors including clinicopathological characteristics and other genes involved in NSCLC. Our results indicated that XAF1 expression was markedly altered in NSCLC tumor samples when compared to that found in normal lung tissues. Predominantly, XAF1 was downregulated in the tumors, except in never-smoker patients. In addition, XAF1 may also be important in the whole cell stress mechanism where the p53 status is crucial.


Rapid screening and early precautions for carbapenem-resistant Acinetobacter baumannii carriers decreased nosocomial transmission in hospital settings: a quasi-experimental study.
Active surveillance has the potential to prevent nosocomial transmission of carbapenem-resistant Acinetobacter baumannii (CRAB). We assessed whether rapid diagnosis using clinical specimen-direct loop-mediated isothermal amplification (LAMP), a rapid molecular diagnostic assay, and subsequent intervention, could reduce CRAB nosocomial transmission in intensive care units (ICUs). A before and after (quasi-experimental) study was conducted in two ICUs at the Mahidol University Faculty of Medicine Ramathibodi Hospital with 3 months of observational period followed by 9 months of interventional period. All patients were screened for CRAB using both the culture and LAMP method from rectal swab and/or bronchial aspirates (intubated patients only) upon admission, weekly thereafter, and upon discharge. During the pre-intervention period, we performed contact precautions based on culture results. In contrast, during the intervention period, we initiated contact precautions within a few hours after sample collection on the basis of LAMP results. A total of 1335 patients were admitted to the ICUs, of which 866 patients (pre-intervention period: 187; intervention period: 679) were eligible for this study. Incidence rate of CRAB infection decreased to 20.9 per 1000 patient-days in the intervention period from 35.2 in the pre-intervention period (P < 0.02). The calculated hazard ratio of CRAB transmission was 0.65 (95% confidence interval [CI], 0.44-0.97). Risk factors for CRAB acquisition included exposure to carbapenem (hazard ratio, 2.54 [95% CI: 1.61-5.57]). LAMP screening for CRAB upon ICU admission proved feasible for routine clinical practice. Rapid screening using LAMP followed by early intervention may reduce CRAB transmission rates in ICUs when compared to conventional intervention.


Removal of Unwanted Hair: Efficacy and Safety of 755-nm Alexandrite Laser Equipped with a 30 mm Spot Handpiece.
Background: Alexandrite 755 nm laser has been currently recognized one of the gold standards for the permanent hair removal business because of its sufficient tissue penetration and higher affinity for melanin compared with Nd:YAG, making the treatment more incisive and effective. Objective: In this study, we evaluated an Alexandrite 755 nm laser with a 30-mm spot for a new and fast hair removal treatment. Methods: Patients of both genders, with skin Fitzpatrick's types, ranging from II to III, were asked to avoid any epilation techniques 4 weeks before the laser hair removal session. Participants underwent a minimum of three to a maximum of eight sessions of treatment, every 6 weeks. Six months after the last treatment, the final evaluation was completed. The treated body areas included arms, legs, axillae, and trunk. The subjects used a numeric scale with a range of 0 to 10 (0: no pain, 10: unbearable), to indicate the associated pain level experienced. By comparing the percentage of terminal hairs before and after laser treatment, hair reduction was determined. Results: The patients reported only slight post-treatment erythema that disappeared after a few hours and only two patients showed persistent erythema lasting a few weeks. The pain was found to be 4.9 ± 0.9 after single pass emission. Conclusions: The 755 nm laser with the new handpiece has proven to be an ideal wavelength for hair removal, allowing efficient, safe, and faster treatments to be performed for the patient.


A multi-target therapeutic potential of Prunus domestica gum stabilized nanoparticles exhibited prospective anticancer, antibacterial, urease-inhibition, anti-inflammatory and analgesic properties.
Phytotherapeutics exhibit diverse pharmacological effects that are based on the combined action of a mixture of phytoconstituents. In this study, Prunus domestica gum-loaded, stabilized gold and silver nanoparticles (Au/Ag-NPs) were evaluated for their prospective anticancer, antibacterial, urease-inhibition, anti-inflammatory, and analgesic properties. Au/Ag-NPs were biosynthesized and characterized with UV-Vis, FTIR, SEM, EDX, and XRD techniques. The effect of gum and metal ion concentration, reaction temperature, and time on the synthetic stability of nanoparticles was studied along with their post-synthetic stability against varying pH and salt concentrations, long-term storage and extremes of temperature. Nanoparticles were tested for anticancer (HeLa cervical cancer cells), antibacterial (Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa), urease inhibition (jack-bean urease), anti-inflammatory (carrageenan-induced paw edema), and antinociceptive (abdominal constriction response) activities. The nanoparticles were mostly spherical with an average particle size between 7 and 30 nm (Au-NPs) and 5-30 nm (Ag-NPs). Au/Ag-NPs maintained their colloidal stability and nanoscale characteristics against variations in physicochemical factors. Au/Ag-NPs have potent anticancer potential (IC50 = 2.14 ± 0.15 μg/mL and 3.45 ± 0.23 μg/mL). Au/Ag-NPs selectively suppressed the growth of S. aureus (10.5 ± 0.6 mm, 19.7 ± 0.4 mm), E. coli (10 ± 0.4 mm, 14.4 ± 0.7 mm), and P. aeruginosa (8.2 ± 0.3 mm, 13.1 ± 0.2 mm), as well as showed preferential inhibition against jack-bean urease (19.2 ± 0.86%, 21.5 ± 1.17%). At doses of 40 and 80 mg/kg, Au-NPs significantly ameliorated the increase in paw edema during the 1st h (P < 0.05, P < 0.01) and 2-5 h (P < 0.001) of carrageenan-induced inflammation compared to the 200 and 400 mg/kg doses of P. domestica gum (P < 0.05, P < 0.001). At similar doses, Au-NPs also significantly abolished (P < 0.01) the tonic visceral, chemically-induced nociception, which was comparable to that of P. domestica gum (200 mg/kg; P < 0.05, 400 mg/kg; P < 0.01).


Finding the true pathway for reversible isomerization of a single azobenzene molecule tumbling on Au(111) surface.
Switchable trans-cis isomerization of azobenzene (AB) and its derivatives on metallic surfaces have offered rich possibilities to functionalize molecular devices. However, the lack of a good understanding of the isomerization pathway has severely limited our ability for rational design. One of the long-debated issues is the cis configuration of the parental AB on the Au(111) surface, for which the experimentally inferred structure differs from the theoretically predicted global minimum. Here, we theoretically identify a new in situ metastable configuration for cis-AB on Au(111) that can reproduce all the observations reported in the scanning tunneling microscopy experiments. It reveals that the bistability of AB on the Au(111) surface is attributed to the significantly increased kinetic stability of the newly discovered cis-AB isomer. A fascinating tumbling pathway that overcomes two energy barriers stimulated by tunneling electrons for the trans-cis AB isomerization on Au(111) has been verified, suggesting a new type of molecular motion based on the AB systems.


Dobutamine stress radionuclide ventriculography reveals silent myocardial dysfunction in Kawasaki disease.
Dobutamine (DOB) stress radionuclide ventriculography (RVG) is proposed for evaluating left ventricular performance in patients with Kawasaki disease (KD). Dobutamine stress RVG, up to 15 microg x kg(-1) x min(-1), was performed in 40 patients with a history of KD, some of whom had a perfusion defect (PD group) on dipyridamole stress thallium-201 myocardial imaging, some of whom had no perfusion defects (NPD group), and some of whom had no coronary artery lesions (C group). No significant differences in either systolic or diastolic indices of the left ventricle at rest were observed between the 3 groups. Although hemodynamic responses were similar in all patients after DOB stress, early diastolic index of the first third filling fraction decreased only in the PD group and was significantly lower in this group compared with the C group (p<0.01). The asynchrony index increased significantly in those patients with coronary stenosis after DOB stress (p<0.05). No serious side-effects were observed during the study. Even late after onset, patients with myocardial ischemia as a result of KD still had impaired early diastolic filling and asynchronous relaxation of the left ventricle. As an alternative to exercise testing, DOB stress RVG is a safe and promising means for serially evaluating left ventricular performance in patients with KD.