A dnaT mutant with phenotypes similar to those of a priA2::kan mutant in Escherichia coli K-12.
The ability to repair damaged replication forks and restart them is important for cell survival. DnaT is essential for replication restart in vitro and yet no definite genetic analysis has been done in Escherichia coli K-12. To begin, dnaT822, an in-frame six-codon (87-92) deletion was constructed. DnaT822 mutants show colony size, cell morphology, inability to properly partition nucleoids, UV sensitivity, and basal SOS expression similar to priA2::kan mutants. DnaT822 priA2::kan double mutants had phenotypes similar to those of the single mutants. DnaT822 and dnaT822 priA2::kan mutant phenotypes were fully suppressed by dnaC809. Previously, a dominant temperature-sensitive lethal mutation, dnaT1, had been isolated in E. coli 15T(-). DnaT1 was found to have a base-pair change relative to the E. coli 15T(-) and E. coli K-12 dnaT genes that led to a single amino acid change: R152C. A plasmid-encoded E. coli K-12 mutant dnaT gene with the R152C amino acid substitution did not display a dominant temperature-sensitive lethal phenotype in a dnaT(+) strain of E. coli K-12. Instead, this mutant dnaT gene was found to complement the E. coli K-12 dnaT822 mutant phenotypes. The significance of these results is discussed in terms of models for replication restart.


Secondary metabolites from Penicillium sp. 8PKH isolated from deteriorated rice straws.
In the search for bioactive secondary metabolites from terrestrial fungi, four compounds, namely, 3-methyl-3H-quinazolin-4-one (1), aurantiomide C (2), 3-O-methylviridicatin (3), and dehydrocyclopeptine (4), were isolated from Penicillium sp. 8PKH, fungal strain, isolated from deteriorated rice straws. The structures of the isolated compounds were identified by extensive NMR and mass analyses and comparison with literature data. This is the first report of the structure of 3-methyl-3H-quinazolin-4-one (1) with full NMR spectral data having been previously identified by GC-MS from Piper beetle. Analysis of the non-polar fractions of the strain extract by GC-MS revealed the presence of additional eight compounds: methyl-hexadecanoate, methyl linoleate, methyl-9 (Z)-octadecenoate, methyl-octadecanoate, cis-9-oxabicyclo (6.1.0) nonane, 9,12-octadecadienal (9E,12E), ethyl-(E)-9-octadecenoate, and 3-buten-2-ol. The isolated compounds were evaluated for their antimicrobial and cytotoxic activities and exhibited little or no inhibitory activities against the test strains. The taxonomical characterisation and fermentation of the fungal strain were reported as well.


[Dentistry oral hygiene and endocarditis. Pathophysiology and prophylactic therapy].
Infectious endocarditis is a cardiac pathology of bacterial, viral or more rarely mycotic origin developing on the surfaces of the endocardium or heart valves. Predisposing conditions are congenital malformations of the heart or valvular acquired alterations, as well as the presence of a valvular prosthesis. The microorganisms involved in the etiology and pathogenesis of the damage of such infection (bacterias, viruses or yeasts) determine the formation of the endocardic vegetations typical of this condition. Such lesions can be located on the valvular or the parietal endocardium and sometimes on the endothelium of a great artery. In despite of the elevated standards of instrumental investigations and therapeutic protocols, the bacterial endocarditis represents a pathology of wide interest, scientific and social, due to its high rate of incidence, morbility and mortality. Still now infectious endocarditis causes death in 20-30% of the patients. Although the significant progress on prevention of the infectious diseases and of the cross infections in dentistry practice, from the tartar ablation up to the oncologic oral surgery, still now the skills of oral hygiene and dentistry represent a potential threat for the development of an infectious endocarditis in predisposed patients. The authors, on the base of the revision of the literature and of their own clinical experience, show the etiology, pathophysiology and the clinical pictures related to such complex disease.


Comparing Post-operative Opioid Consumption before and after a Patient-Controlled Analgesia Shortage: A Re-evaluation of Safety and Effectiveness.
This retrospective cohort study aimed to compare post-surgical opioid consumption before and after a PCA (patient-controlled analgesia) shortage. The study evaluated patients who received PCA vs. nurse-administered opioid analgesia (non-PCA). Two hundred and twenty-four patients ≥18 years who were initiated on analgesia within 24 h of surgery were included. The primary outcome was opioid consumption in average daily oral morphine milliequivalents (MME). The results showed that patients in the PCA group had increased MME consumption (162 ± 100.4 vs. 70.7 ± 52.8, p < 0.01), increased length of hospital stay (4.2 vs. 3.2 days, p < 0.01), and increased frequency of nausea (33 vs. 17.9%, p < 0.01). After controlling for confounding factors, the PCA group utilized significantly more opioids (84.6 MME/day, p < 0.01) than the non-PCA group. There was no difference in pain AUC/T (0.19 ± 0.07 vs. 0.21 ± 0.08, p = 0.07) and average opioid prescribing upon discharge (150 [77.5-360] vs. 90 [77.5-400], p = 0.64) between the PCA group and non-PCA group, respectively. These results question the routine use of PCA in post-operative patients due to the increased risk of opioid consumption, longer length of hospital stay, and higher incidence of nausea.


Suppression of cortisol responses to exogenous adrenocorticotrophic hormone, and the occurrence of side effects attributable to glucocorticoid excess, in cats during therapy with megestrol acetate and prednisolone.
The major purpose of this investigation was to determine the effect of prednisolone and megestrol acetate in cats on the adrenal cortisol response to exogenous adrenocorticotrophic hormone during drug administration at dose rates employed for management of some inflammatory feline dermatoses. Prednisolone (at least 2 mg/kg/day) and megestrol acetate (5 mg/cat/day) were each administered orally to seven cats from days 1 to 16. Three additional cats received no therapy. Basal and stimulated cortisol concentrations, food and water intake, hematology, blood biochemistry, urinalyses, and hepatic and cutaneous histology were studied in all cats before, during, and two weeks following the end of treatment. Cats given prednisolone or megestrol acetate had significant suppression of stimulated cortisol levels on day 8. This change was more marked on day 15, when the suppression in cats given megestrol acetate was also significantly more severe than in those receiving prednisolone. Recovery of adrenal reserve was considered present on day 30 in six of seven cats given prednisolone, but in only three of seven receiving megestrol acetate. Eosinopenia, glycosuria and hepatocyte swelling from glycogen deposition were occasionally recorded in treated cats of both groups, providing additional circumstantial evidence for glucocorticoid activity of megestrol acetate in cats. It is advised that abrupt withdrawal of prednisolone or megestrol acetate therapy be avoided in this species to reduce the chance of precipitating clinical signs of hypoadrenocorticism, even after treatment for as little as one week.


Human CD4+CD45R0+ and CD4+CD45RA+ T cells synergize in response to alloantigens.
Alloantigens, unlike recall antigens, activate both CD45RA+ (naive) and CD45R0+ (memory) CD4+ cells to the same extent. These T cell subsets may therefore interact with each other in response to alloantigens on transplanted grafts. We have investigated if the ability of activated CD4+CD45RA+ and CD4+CD45R0+ T cells to produce and respond to interleukin 2 (IL2) and IL4 may be involved in this interaction. After activation, both subsets up-regulate their IL2 receptor (IL2R) and IL4R expression, yet IL4 substantially enhanced the proliferation of the CD4+CD45RA+ but not of the CD4+CD45R0+ T cell subset, while IL2 increased the proliferation of CD4+CD45R0+ but not of the CD4+CD45RA+ T cells. Significantly, the CD4+CD45RA+ T cells synthesized two- to threefold more mRNA for IL2 than the CD4+CD45R0+ subset, while the CD4+CD45R0+ T cells synthesized mRNA for IL4 and interferon-gamma exclusively. The addition of IL2 to alloactivated CD4+CD45R0+ T cells further up-regulated their production of all three lymphokine mRNA; in contrast, IL4 induced an increase in mRNA for IL2 in only the alloactivated CD4+CD45RA+ subset. The reciprocity in the ability of both these CD4+ T cells to synthesize and respond to IL2 and IL4 may provide a rationale for the regulation of lymphokine interactions in vivo. Furthermore, the synergy between these subsets in response to alloantigens, which was directly quantitated by co-culturing CD4+CD45RA+ and CD4+CD45R0+ cells together prior to activation, may potentiate the alloreactivity against transplanted grafts in vivo.


Hydrogen abstraction from n-butanol by the methyl radical: high level ab initio study of abstraction pathways and the importance of low energy rotational conformers.
Hydrogen abstraction reactions by the methyl radical from n-butanol have been investigated at the ROCBS-QB3 level of theory. Reaction energies and product geometries for the most stable conformer of n-butanol (ROH) have been computed, the reaction energies order α < γ < β < δ < OH. The preference for n-butane to favour H-abstraction at C(β) and C(γ) while, in contrast, n-butanol favours radical reactions at the C(α) carbon is rationalised. Transition state (TS) barriers and geometries for the most stable conformer of n-butanol are presented, and discussed with respect to the Hammond postulate. The reaction barriers order as α < OH < γ < β < δ. This relative ordering is not consistent with product radical stability, C-H bond dissociation energies or previous studies using O[combining dot above]H and HO[combining dot above](2) radicals. We provide a molecular orbital based rationalisation for this ordering and answer two related questions: Why is the γ-channel more stable than the β-channel? Why do the two C(γ)-H H-abstraction TS differ in energy? The method and basis set dependence of the TS barriers is investigated. The Boltzmann probability distribution for the n-butanol conformers suggests that low energy conformers are present in approximately equal proportions to the most stable conformer at combustion temperatures where ĊH(3) radicals are present. Thus, the relative significance of the various H-abstraction channels has been assessed for a selection of higher energy conformers (ROH'). Key results include finding that higher energy n-butanol conformers (E(ROH') > E(ROH)) can generate lower energy product radicals, E(ROH') < E(ROH). Moreover, higher energy conformers can also have a globally competitive TS energy for H-abstraction.


Decision-making deficits in obsessive-compulsive disorder are associated with abnormality of recency and response consistency parameter in prospect valence learning model.
Patients with obsessive-compulsive disorder (OCD) have deficits in decision-making in the Iowa Gambling Task (IGT). However, no study has investigated the parameters of the prospect valence learning (PVL) model in the IGT for OCD. This study aimed to investigate deficits in decision-making in OCD using the PVL model and identify whether the parameters of the PVL model were associated with obsessive-compulsive severity. Forty-seven medication-free patients with OCD were compared with 47 healthy controls (HCs). Decision-making was measured using the total net and block net scores of the IGT. A PVL model with a decay-reinforcement learning rule (PVL-DecayRI) was used to investigate the parameters of the model. Correlation analysis was conducted between each parameter of the PVL-DecayRL and obsessive-compulsive symptoms. The total net score of patients with OCD was significantly lower than that of the HCs. The block net scores of the OCD group did not differ across the five blocks, whereas in the HCs, the fifth block net score was significantly higher than the block net scores of the first and second blocks. The values of the recency and response consistency parameters of the PVL-DecayRI in patients with OCD were significantly lower than those in HCs. The recency parameter positively correlated with the Y-BOCS obsessive score. Meanwhile, there was no correlation between consistency parameter values and symptom severity in OCD. Our detailed analysis of the decision-making deficit in OCD suggests that the most recent outcome has a small influence on the expectancy of prospect valence, as indicated by the lower recency parameter, and is characterized by more impulsive choices, as indicated by the lower consistency parameter.


A nomogram-based optimized Radscore for preoperative prediction of lymph node metastasis in patients with cervical cancer after neoadjuvant chemotherapy.
To construct a superior single-sequence radiomics signature to assess lymphatic metastasis in patients with cervical cancer after neoadjuvant chemotherapy (NACT). The first half of the study was retrospectively conducted in our hospital between October 2012 and December 2021. Based on the history of NACT before surgery, all pathologies were divided into the NACT and surgery groups. The incidence rate of lymphatic metastasis in the two groups was determined based on the results of pathological examination following lymphadenectomy. Patients from the primary and secondary centers who received NACT were enrolled for radiomics analysis in the second half of the study. The patient cohorts from the primary center were randomly divided into training and test cohorts at a ratio of 7:3. All patients underwent magnetic resonance imaging after NACT. Segmentation was performed on T1-weighted imaging (T1WI), T2-weighted imaging, contrast-enhanced T1WI (CET1WI), and diffusion-weighted imaging. The rate of lymphatic metastasis in the NACT group (33.2%) was significantly lower than that in the surgery group (58.7%, P=0.007). The area under the receiver operating characteristic curve values of Radscore_CET1WI for predicting lymph node metastasis and non-lymphatic metastasis were 0.800 and 0.797 in the training and test cohorts, respectively, exhibiting superior diagnostic performance. After combining the clinical variables, the tumor diameter on magnetic resonance imaging was incorporated into the Rad_clin model constructed using Radscore_CET1WI. The Hosmer-Lemeshow test of the Rad_clin model revealed no significant differences in the goodness of fit in the training (P=0.594) or test cohort (P=0.748). The Radscore provided by CET1WI may achieve a higher diagnostic performance in predicting lymph node metastasis. Superior performance was observed with the Rad_clin model.


Domestication of newly evolved hexaploid wheat-A journey of wild grass to cultivated wheat.
Domestication of wheat started with the dawn of human civilization. Since then, improvement in various traits including resistance to diseases, insect pests, saline and drought stresses, grain yield, and quality were improved through selections by early farmers and then planned hybridization after the discovery of Mendel's laws. In the 1950s, genetic variability was created using mutagens followed by the selection of superior mutants. Over the last 3 decades, research was focused on developing superior hybrids, initiating marker-assisted selection and targeted breeding, and developing genetically modified wheat to improve the grain yield, tolerance to drought, salinity, terminal heat and herbicide, and nutritive quality. Acceptability of genetically modified wheat by the end-user remained a major hurdle in releasing into the environment. Since the beginning of the 21st century, changing environmental conditions proved detrimental to achieving sustainability in wheat production particularly in developing countries. It is suggested that high-tech phenotyping assays and genomic procedures together with speed breeding procedures will be instrumental in achieving food security beyond 2050.


C57BL/6 congenic mouse NRASQ61K melanoma cell lines are highly sensitive to the combination of Mek and Akt inhibitors in vitro and in vivo.
RAS is frequently mutated in various tumors and known to be difficult to target. NRASQ61K/R are the second most frequent mutations found in human skin melanoma after BRAFV600E . Aside from surgery, various approaches, including targeted therapies, immunotherapies, and combination therapies, are used to treat patients carrying NRAS mutations, but they are inefficient. Here, we established mouse NRASQ61K melanoma cell lines and genetically derived isografts (GDIs) from Tyr::NRASQ61K mouse melanoma that can be used in vitro and in vivo in an immune-competent environment (C57BL/6) to test and discover novel therapies. We characterized these cell lines at the cellular, molecular, and oncogenic levels and show that NRASQ61K melanoma is highly sensitive to the combination of Mek and Akt inhibitors. This preclinical model shows much potential for the screening of novel therapeutic strategies for patients harboring NRAS mutations that have limited therapeutic options and resulted in poor prognoses.


18F-FDG-PET/CT in the evaluation and differential diagnosis of active large-vessel vasculitis. A prospective study.
Introduction: Large-vessel vasculitis has non-specific clinical symptoms, which can delay the diagnosis. Early recognition and treatment of the disease can help to avoid late complications. 18 F-FDG-PET can detect the inflammation of the vessel wall in the early stage of the disease with high sensitivity. CT is used to localize vasculitis. Aim: To examine the performance of 18F-FDG-PET/CT in patients with suspected large-vessel vasculitis, during relapse and remission, focusing on disease activity and extent. Method: 43 patients were evaluated. They were classified according to the clinical questions: steroid-naive suspected vasculitis, suspected vasculitis on steroid treatment, patients with relapse and in remission. We examined 10 cancer patients in control. We carried out visual and quantitative analysis of the 18F-FDG uptake of vessel walls. During quantitative evaluation, we determined standardised uptake values (SUVmax) of vessel wall segments compared to liver. Results: We found active disease in 5 patients examined for primary diagnosis, moreover, in 5 patients with relapse. The disease involved 3 or more vessel segments in fifty percent of the active cases. In the visually active group, the SUVmax was significantly lower in patients on steroid treatment than in steroid-naive cases (1.17 ± 0.11 vs. 1.43 ± 0.29; p = 0.005). We confirmed remission in 2 cases after therapy. In the inactive group, we found other types of inflammatory disorders in 8 cases. Conclusion: 18F-FDG-PET/CT is an effective diagnostic tool for large-vessel vasculitis, and can be used to determine the activity and extent of the disease. Steroid treatment influences the 18F-FDG-uptake of vessel wall. Orv Hetil. 2020; 161(20): 829-838.


Severe Generalized Epidermolysis Bullosa Simplex in Two Hong Kong Children due to De Novo Variants in KRT14 and KRT5.
We report two Hong Kong children with severe generalized epidermolysis bullosa simplex (EBS), the most severe form of EBS, without a family history of EBS. EBS is a rare genodermatosis usually inherited in an autosomal dominant fashion although rare autosomal recessive cases have been reported. Genetic studies in these patients showed that the first case was due to a novel de novo heterozygous variant, c.377T>G (NM_000526.5 (c.377T>G, p.Leu126Arg)) in the KRT14 gene and the second case was due to a rare de novo heterozygous variant c.527A>G (NM_000424.4, c.527A>G, p.Asn176Ser) in the KRT5 gene. To our knowledge, the c.377T>G variant in the KRT14 gene has not been previously reported, and the c.527A>G variant in the KRT5 gene is a rare cause of severe generalized EBS. In severe generalized EBS, infants exhibit severe symptoms at the onset; however, they tend to improve with time. A precise genetic diagnosis in these two cases aided in counseling the families concerning the prognosis in their affected children and the recurrence risk for future pregnancies.


[Treatment of tuberculosis infection].
Tuberculosis is a chronic bacterial infection caused by Mycobacterium tuberculosis. It continues as an important cause of morbidity and mortality worldwide, especially in impoverished countries and where human immunodeficiency virus infection is endemic. The modern treatment of tuberculosis is based on the administration of effective drugs. Regimens do not differ for pulmonary and extra-pulmonary tuberculosis. In order to prevent the emergence of drug-resistant organisms, which is present initially in very small numbers, at least two effective drugs are always required. Short-course therapy has been developed to mitigate the consequences of patient default. It is best considered as consisting of two phases. An initial 2-month intensive phase of daily therapy should include isoniazid, rifampin, ethambutol and pyrazinamide. A consolidation phase of daily therapy with isoniazid and rifampin should be continued for an additional 4 months, preferably more for special clinical circumstances. This standardized drug strategy is successful only if the resources conserved by shortening treatment are used to maintain patient compliance. Completely supervised regimens have been also developed with success. Defaults lead not only to treatment failure but also the emergence and transmission of drug-resistant organisms. Treatment of confirmed or suspected drug-resistant tuberculosis is difficult and should only be made on experts consultation. More difficult to use and/or less effective than first line drugs, second line drugs could be chosen and associated. However drug toxicities should be monitored, with greatest concern to hepatitis. Follow-up of patients must be organised until 2 years after the completion of therapy to detect relapses. Treatment includes prophylactic measures which are a major modality for decreasing the spread of infection.


Switch-ON Near IR Fluorescent Dye Upon Protonation: Helically Twisted Bis(Boron Difluoride) Complex of π-Extended Corrorin.
A novel helically twisted π-extended corrorin derivative having two boron dipyrrin (BODIPY) moieties at the periphery, a BODIPY DYEmer (6-BF2 ) cross-bridged with π-conjugated dipyrrinylidene unit, was synthesized and characterized. The neutral 6-BF2 is nonfluorescent due to the internal photoinduced charge transfer (CT) character in the excited state as inferred from the femtosecond transient absorption spectroscopy. However, upon protonation, the CT process is suppressed and the species H6-BF2+ becomes near infrared (IR) emissive. With the aid of rigid helical conformations in 6-BF2 , the helical isomers (P- and M-forms) were resolved and their chiroptical property was investigated. The distinct circular dichroism (CD) spectral changes of the enantiomers were observed in the presence of acids, which demonstrates that 6-BF2 can be utilized for potential acid-responsive chiroptical materials.


The NIF x-ray spectrometer calibration campaign at Omega.
The calibration campaign of the National Ignition Facility X-ray Spectrometer (NXS) was carried out at the Omega laser facility. Spherically symmetric, laser-driven, millimeter-scale x-ray sources of K-shell and L-shell emission from various mid-Z elements were designed for the 2-18 keV energy range of the NXS. The absolute spectral brightness was measured by two calibrated spectrometers. We compare the measured performance of the target design to radiation hydrodynamics simulations.


Distinctive Subpopulations of Stromal Cells Are Present in Human Lymph Nodes Infiltrated with Melanoma.
Metastasis of human tumors to lymph nodes (LN) is a universally negative prognostic factor. LN stromal cells (SC) play a crucial role in enabling T-cell responses, and because tumor metastases modulate their structure and function, this interaction may suppress immune responses to tumor antigens. The SC subpopulations that respond to infiltration of malignant cells into human LNs have not been defined. Here, we identify distinctive subpopulations of CD90+ SCs present in melanoma-infiltrated LNs and compare them with their counterparts in normal LNs. The first population (CD90+ podoplanin+ CD105+ CD146+ CD271+ VCAM-1+ ICAM-1+ α-SMA+) corresponds to fibroblastic reticular cells that express various T-cell modulating cytokines, chemokines, and adhesion molecules. The second (CD90+ CD34+ CD105+ CD271+) represents a novel population of CD34+ SCs embedded in collagenous structures, such as the capsule and trabeculae, that predominantly produce extracellular matrix. We also demonstrated that these two SC subpopulations are distinct from two subsets of human LN pericytes, CD90+ CD146+ CD36+ NG2- pericytes in the walls of high endothelial venules and other small vessels, and CD90+ CD146+ NG2+ CD36- pericytes in the walls of larger vessels. Distinguishing between these CD90+ SC subpopulations in human LNs allows for further study of their respective impact on T-cell responses to tumor antigens and clinical outcomes.


A comparison of hypothalamically induced biting attack with natural predatory behavior in the cat.
A comparison of biting attack induced by electrical stimulation of the cat hypothalamus with natural feline predatory attack revealed a high degree of similarity between the two behaviors. All major components of natural attack could be elicited by hypothalamic stimulation in cats not showing spontaneous attack, including stalking approach to prey, holding and pinning of the prey with forepaws, biting directed to the head and neck, prey kicking, prey carrying, and prey eating. Differences between natural and elicited attack appeared to be related to the range of completenes of elicited attack, the greater intensity of elicited attack, and the presence of unrelated responses induced by hypothalamic stimulation. Results support the view that hypothalamic stimulation facilitates sensorimotor mechanisms for separate response components, which are highly sensitive to specific stimulus features of the environment and potential goal objects.


Successful planned pregnancy in a patient with St Jude medical prosthetic mitral valve.
A primiparous woman with a St Jude Medical (SJM) mitral valve prosthesis and receiving anticoagulant drug delivered a healthy male infant by spontaneous normal vaginal delivery. Oral warfarin was replaced by subcutaneous heparin at the 6th week of gestation and the dosage of heparin was adjusted to maintain a partial thromboplastin time between 2.0 and 2.5 times the control value. Treatment with heparin was administered during the first trimester and the last 5 weeks of gestation. Warfarin was used between the 13th and 32nd week. Successful pregnancy and delivery in patients with a SJM mitral valve prostheses is possible with careful maintenance of anticoagulation.


Detection by near-infrared spectroscopy of large lipid cores at culprit sites in patients with non-ST-segment elevation myocardial infarction and unstable angina.
This study was performed to assess the lipid burden of culprit lesions in non-ST-segment elevation myocardial infarction (non-STEMI) and unstable angina (UA). A recent intracoronary near-infrared spectroscopy (NIRS) study showed 85% of STEMI culprit lesions have a maximum lipid core burden index in 4-mm (maxLCBI(4mm)) ≥ 400. Whether culprit lesions in non-STEMI and UA are characterized by a similarly large lipid burden is unknown. We studied 81 non-STEMI and UA patients undergoing culprit vessel NIRS imaging before stenting. Culprit segments were compared to all nonoverlapping 10-mm nonculprit segments for maxLCBI(4mm). Culprit segments in non-STEMI and UA were compared for the frequency of maxLCBI(4mm) ≥ 400. Among 81 patients (53.1% non-STEMI, 46.9% UA), non-STEMI culprit segments had a 3.4-fold greater maxLCBI(4mm) than nonculprits (448 ± 229 vs 132 ± 154, P < 0.001) and UA culprit segments had a 2.6-fold higher maxLCBI(4mm) than nonculprits (381 ± 239 vs 146 ± 175, P < 0.001). NIRS detected a maxLCBI(4mm) ≥ 400 in 63.6% of culprit segments in NSTEMI and in 38.5% of culprit segments in UA (P = 0.02). Against a background of nonculprit segments, maxLCBI(4mm) ≥ 400 had a sensitivity of 63.6% and specificity of 94.0% for culprit segments in NSTEMI and a sensitivity of 38.5% and specificity of 89.8% for culprit segments in UA. Large lipid cores similar to those recently detected by NIRS at STEMI culprit sites were frequently observed at culprit sites in patients with non-STEMI and UA. These findings support ongoing prospective trials designed to determine if NIRS can provide site-specific prediction of future acute coronary events.


Combined Training of Candidates, Scholars, And Psychotherapists: A Model of Psychoanalytic Education for the Twenty-First Century.
A new model of psychoanalytic education is proposed that will meet the challenges of educating candidates in a new century. Prospective candidates have varying opinions about the value of analytic training, opinions that reflect economic and cultural conditions different from those facing previous generations. Overall, today's graduate-level students hold less favorable attitudes toward psychoanalysis than did their counterparts in the past. The proposed model calls for combining analytic candidates, psychotherapy students, and academic scholars for two years in a Psychoanalytic Studies Program (PSP), after which candidates take their subsequent years of training in a cohort made up exclusively of analytic candidates. A curriculum that focuses on the core concepts in psychoanalysis allows students in all three categories to learn the foundational knowledge of psychoanalysis that once was widely taught in graduate mental health programs. The philosophy that underlies the model and the structure and orientation of the course sequences are presented. Implementatiion of the model having shown positive results, its strengths and limitations are evaluated against the traditional model, in which candidates and psychotherapy students are educated separately.


Clinical trial/experimental study (consort compliant): Optimal time period to achieve the effects on synbiotic-controlled wheezing and respiratory infections in young children.
Urban life is often followed by immune dysfunction and loss of immune tolerance in the youngest children. The study aimed to determine optimal time efficiency of a synbiotic (5 x 10⁹ Lactobacillus acidophilus Rosell-52, Bifidobacterium infantis Rosell-33, Bifidobacterium bifidum Rosell-71) in controlling respiratory infections and wheezing disease. We randomly selected a group of children younger than five years, hospitalized earlier, and classified them into three groups. The incidence of respiratory infection before the study was once a month, while after a three-month supplementation with the synbiotic children rarely suffered from respiratory infections, and the state was maintained after six-month and nine-month supplementations with the synbiotic. The decreased incidence of respiratory infections was followed by a falling incidence of concomitant wheezing. A significant increase in tIgA serum was observed in all groups for only three months, the increase being the highest in children with recurrent respiratory infections accompanied by wheezing. After a nine-month administration of the synbiotic, total IgE serum was lower in all groups of patients. The optimal duration of administration of the synbiotic containing three probiotic cultures to provide effective control of the frequency of respiratory infections was three months, and six months were required to establish control of the frequency of wheezing. This synbiotic is useful for immunomodulation in children and is well-tolerated in young children.


Flow Dynamic Pattern in Liver and Renal Transplantation under Exercise Prescription Program.
Cardiovascular diseases in the context of renal and liver transplants remain the leading cause of morbidity and mortality. Physical exercise at a moderate intensity is allowed to contrast the risk profile. Echocardiographic evaluation is essential to stratifying potential cardiotoxicity by the standard and, more recently, the deformation and dynamic study of the intracardiac vortex. This study aims to investigate the vortex echo parameters of solid-organ-transplanted subjects who are physically active compared to a control group of healthy subjects. A group of 33 transplanted subjects (16 kidneys and 17 livers) was studied via a transthoracic echocardiography exam, comprehending the myocardial deformation parameters of global longitudinal strain (GLS), twisting of the left ventricle (LV) chamber, and HyperDoppler image acquisition. The subjects enrolled in this study were 50 in total: there were 33 transplanted and 17 healthy subjects. The transplanted subjects presented higher values of interventricular septum in diastole (IVSd p = 0.001), posterior wall diastolic (PWd p = 0.05), and left ventricle mass index (LVMI p = 0.029); ejection fraction (EF) was found to be higher in athletes (p < 0.001). Transplanted subjects presented mild diastolic dysfunction, emerging only from septal E values (p = 0.001). The 4DStrain (p = 0.018) and GLS2c (p = 0.017) were significantly better in the athletes. All of the geometrical and energetical vortex data were in the normal range and no significant differences were found. An interesting positive correlation was evident for the diastolic parameter, particularly the E/A ratio (p = 0.023) and E' septal value (p = 0.049), along with the vorticity fluctuation. This behavior was present for all subjects, particularly those that were transplanted (p = 0.005). In the vortex investigation, especially in cases of normal EF, the positive correlation of some diastolic parameters with the flow dynamic patterns corroborates this hypothesis. The HyperDoppler analysis could be helpful to detecting potential damage earlier in the diastolic time before a systolic deficiency.


Hairpin formation in d-AAGCTTAAGCTT under high salt conditions shows unusual properties.
The hairpin-duplex equilibria of the dodecamer d-AAGCTTAAGCTT and interaction of the duplex form with a pentapeptide, KGWGK, has been studied. UV thermal transitions are monophasic at low salt but biphasic at higher salt concentrations. At 10(-5) M or less oligomer concentration biphasic melting curves persist till 900 mM NaCl. The d(Tm)/d log(Na+) for the duplex form is 12 degrees C and for the hairpin is 18 degrees C. The delta H and delta S values for duplex formation are low (-25 K cal/mole and -59 Cal/mole respectively). KGWGK binds to the duplex form with a binding constant K = 3.4 x 10(5)M-1 measured from fluorescence quenching of tryptophan. These unusual results are markedly different from that reported for d-AGATCTAGATCT (Biochemistry 31, 6241-6245) and are discussed in terms of sequence dependence of loop folding and cruciform extrusion pathway of hairpin formation.


Toxic Neuropathies.
The purpose of this article is to provide an overview and update on the most clinically relevant toxic neuropathies. Broadly, toxic neuropathies were previously quite rare with the notable exception of neuropathy from alcohol or older chemotherapeutics. The development of newer therapies, particularly immunotherapy to treat malignancy, has resulted in a substantial increase in the occurrence of toxic neuropathies that require timely recognition and treatment. The understanding of other toxic neuropathies continues to evolve, such as statin-induced neuropathy, which new evidence suggests is much less common than previously suspected. Toxic neuropathies can be caused by medications, supplements, and recreational substances that injure peripheral nerves. Medications have evolved in the past 2 decades, as have the types of neuropathies that can be seen as related toxicities. In some areas of medicine, new classes and generations of drugs are associated with a lower incidence of toxic neuropathy.


Note: A compact three-axis optical force/torque sensor using photo-interrupters.
By integrating four photo-interrupters in a cross-shaped structure, we developed a compact three-axis optical force/torque (F/T) sensor. The developed sensor has a diameter of 28 mm and a thickness of 7 mm. Despite simplicity and compactness, the experiments with a prototype of the proposed sensor demonstrate notably high accuracy. The RMS errors are 0.5% ± 0.1% of the maximum vertical force in z-axis, 1.9% ± 0.2% of the maximum torque in x-axis, and 2.0% ± 0.3% of the maximum torque in y-axis. It is expected that the proposed sensor allows cost-effective integration of robot systems requiring compact and multi-axis F/T sensors such as a walking assist robot.


Tight Junction Protein Occludin Is a Porcine Epidemic Diarrhea Virus Entry Factor.
Porcine epidemic diarrhea virus (PEDV), the causative agent of porcine epidemic diarrhea, has caused huge economic losses in pig-producing countries. Although PEDV was long believed to replicate in the intestinal epithelium by using aminopeptidase N as a receptor, the mechanisms of PEDV infection are not fully characterized. In this study, we found that PEDV infection of epithelial cells results in disruption of the tight junctional distribution of occludin to its intracellular location. Overexpression of occludin in target cells makes them more susceptible to PEDV infection, whereas ablation of occludin expression by use of small interfering RNA (siRNA) in target cells significantly reduces their susceptibility to virus infection. However, the results observed with occludin siRNA indicate that occludin is not required for virus attachment. We conclude that occludin plays an essential role in PEDV infection at the postbinding stages. Furthermore, we observed that macropinocytosis inhibitors blocked occludin internalization and virus entry, indicating that virus entry and occludin internalization are closely coupled. However, the macropinocytosis inhibitors could not impede virus replication once the virus had entered host cells. This suggests that occludin internalization by macropinocytosis or a macropinocytosis-like process is involved in the virus entry events. Immunofluorescence confocal microscopy showed that PEDV was trapped at cellular junctional regions upon macropinocytosis inhibitor treatment, indicating that occludin may serve as a scaffold in the vicinity of virus entry. Collectively, these data show that occludin plays an essential role in PEDV infection during late entry events. Our observation may provide novel insights into PEDV infection and related pathogenesis.IMPORTANCE Tight junctions are highly specialized membrane domains whose main function is to attach adjacent cells to each other, thereby forming intercellular seals. Here we investigate, for the first time, the role of the tight junction protein occludin in PEDV infection. We observed that PEDV infection induced the internalization of occludin. By using genetic modification methods, we demonstrate that occludin plays an essential role in PEDV infection. Moreover, PEDV entry and occludin internalization seem to be closely coupled. Our findings reveal a new mechanism of PEDV infection.


The role of two Pseudomonas aeruginosa anthranilate synthases in tryptophan and quorum signal production.
Pseudomonas aeruginosa is a Gram-negative, opportunistic pathogen that causes infections in the lungs of individuals with the genetic disease cystic fibrosis. Density-dependent production of toxic factors regulated by the Pseudomonas quinolone signal (2-heptyl-3-hydroxy-4-quinolone; PQS) have been proposed to be involved in P. aeruginosa virulence. PQS biosynthesis requires conversion of the central metabolite chorismate to anthranilate by anthranilate synthase. This reaction is also the first step in tryptophan biosynthesis. P. aeruginosa possesses two functional anthranilate synthases, TrpEG and PhnAB, and these enzymes are not functionally redundant, as trpEG mutants are tryptophan auxotrophs but produce PQS while mutants in phnAB are tryptophan prototrophs but do not produce PQS in minimal media. The goal of the work described in this paper was to determine the mechanism for this lack of functional complementation of TrpEG and PhnAB. Our results reveal that overexpression of either enzyme compensates for tryptophan auxotrophy and PQS production in the trpEG and phnAB mutants respectively, leading to the hypothesis that differential regulation of these genes is responsible for the lack of functional complementation. In support of this hypothesis, trpEG was shown to be expressed primarily during low-density growth while phnAB was expressed primarily at high density. Furthermore, dysregulation of phnAB expression eliminated tryptophan auxotrophy in the P. aeruginosa trpEG mutant. Based on these data, we propose a model for anthranilate sequestration by differential transcriptional regulation of the two P. aeruginosa anthranilate synthase enzymes.


Genetically encoded bright Ca2+ probe applicable for dynamic Ca2+ imaging of dendritic spines.
G-CaMP is a Ca2+ probe based on a single green fluorescent protein (GFP). G-CaMP shows a large fluorescence increase upon Ca2+ binding, but its fluorescence is dim and pH sensitive, similar to other single GFP-based probes. Here we report an improved G-CaMP, named G-CaMP1.6, which enables easier detection of intracellular Ca2+ signals. G-CaMP1.6 was approximately 40 times more fluorescent than G-CaMP, mainly due to an increase in quantum yield. Furthermore, compared with G-CaMP, G-CaMP1.6 had not only a lower pH sensitivity but also a higher selectivity for divalent cations having an ionic radius similar to Ca2+. Ca2+ sensitivity of G-CaMP1.6 (Kd = 146 nM, Hill coefficient = 3.8, Fmax/Fmin = 4.9) was slightly shifted toward higher affinity compared with that of G-CaMP. When expressed in mammalian cells, G-CaMP1.6 showed large fluorescence changes with drug applications. Notably, local Ca2+ changes in such tiny structures as dendritic spines of neurons were successfully observed with G-CaMP1.6, this being the first observation using a GFP-based probe. Additional mutations in Ca2+-binding sites of G-CaMP1.6 shifted the affinity for Ca2+ and reduced the Ca2+-buffering effect. G-CaMP1.6-CaM(E140K), which has a mutation in the Ca2+ binding site, is an improved probe with its increased brightness and reduced Ca2+-buffering capacity.


Effect of indomethacin on bone remodelling in rabbit ear chambers.
The effect of indomethacin medication on bone tissue resorption and formation was assessed by qualitative and quantitative microscopy in vivo in 18 rabbit ear chambers in 12 rabbits. Five bone tissue experiments were carried out during half a year using the rabbits as their own controls. One daily oral indomethacin dosage of 10 mg/kg body weight given for 3 weeks inhibited resorption of bone autografts in this type of vital chamber and thus inhibited disordered osteoclastic activity. The effect of indomethacin on ordered osteoclastic activity was not quantitated, nor was its effect on parallel-fibred bone tissue formation assessed. The same dosage of indomethacin clearly retarded woven-fibred bone tissue formation (by 1 week) as compared to control experiments, in which the chambers were used as their own controls. It is concluded that indomethacin treatment of patients liable to develop para-articular ossifications therefore seems rational. Indomethacin and drug-induced arthropathy of the hip-joint is discussed in relation to the presented results. A final appraisal of the role of indomethacin in this complex process must await additional material.


Estimated dietary isoflavone intake among Korean adults.
This study estimated the isoflavone intake level in Koreans using Food Frequency Questionnaire and analyzed related variables. The results showed that the average daily intake of isoflavone in adults was shown as 23.1 mg. The isoflavone intake level at 50 percentile was 16.9 mg (0~190 mg), and 10% of adults took almost 50 mg of isoflavone a day and 10% took about 5 mg a day. The major food sources for isoflavone in Koreans were in the order of soybean, soybean paste, soy milk, soybean curd (tofu), and bean sprouts; the intake was different depending on age, educational background, occupation, economic standard, and family type. The result showed higher isoflavone intake levels in the group over 30 years old and the highest isoflavone intake in subjects working in farming/fishery, followed by housemakers. According to the differences by families the families with elderly members showed 50% higher isoflavone intake than young families with friends or siblings. Depending on related ecological variables, therefore, various nutrition education programs should be developed for a variety of intakes of soybean foods, along with easy and simple cooking methods as parts of continuous research.


Sulphasalazine treatment in protracted familial Mediterranean fever arthritis.
Familial Mediterranean fever (FMF) is an autosomal recessive disease characterized by self-limited attacks of fever and polyserositis. Articular involvement in early-onset FMF is a common finding characterized by non-erosive, generally asymmetric monoarthritis in large joints. Protracted FMF arthritis was reported in 2.6% of Turkish patients. An 8-year-old female who has a history of FMF for 5 years applied to our hospital with complaints of persistent swelling and pain of her left knee for 8 months. The patient had been tried to be managed with non-steroidal anti-inflammatory drugs as well as intra-articulary steroids and colchicine. However, arthritis and acute phase response persisted. With sulphasalazine, complete recovery was achieved. It is our belief that sulphasalazine can be a choice of medical treatment in protracted FMF arthritis.


[A case of malakoplakia of the kidney and urinary bladder].
A case of malakoplakia which co-existed in the kidney and the urinary bladder is reported. The patient was a 51-year-old female who had been suffering from asymptomatic macroscopic hematuria. Cystoscopic examination revealed multiple yellow-red nodules in the posterior and lateral wall of the bladder. Transurethral punch biopsy of the mucosa was performed and histopathological examination of the specimens revealed the diagnosis of malakoplakia. The laboratory data at admission were within normal limits except for slightly accelerated erythrocyte sedimentation rate. The culture of urine was negative. The cytology of urine was interpreted as Papanicolaou class I. Intravenous pyelography demonstrated filling defect of the left upper renal calyx, which was confirmed by retrograde pyelography. Computerized tomography showed dilatation of the left upper calyx. An exploratory operation disclosed that the upper calyceal neck was obstructed by several small nodular lesion, which were very similar to the lesions in the bladder. Because intraoperative pathological diagnosis of the frozen section was malakoplakia, partial nephrectomy was performed. Histopathological examination of the kidney and the bladder showed typical Michaelis-Gutmann bodies in the granulomatous lesions. Follow-up examinations 4 months after operation revealed some residual lesions in the bladder which were treated with bethanechol, but no recurrence in the kidney.


Deep brain stimulation in an adolescent with hypomyelination with atrophy of the basal ganglia and cerebellum due to a TUBB4A mutation: illustrative case.
Hypomyelination with atrophy of the basal ganglia and cerebellum (H-ABC) is a rare genetic disease due to a TUBB4A mutation, with motor features including dystonia. Deep brain stimulation (DBS) can be used to treat dystonia in pediatric populations, although the response is highly variable and preferential toward specific etiologies. A single pediatric subject with H-ABC received DBS using a staged procedure involving temporary depth electrode placement, identification of optimal stimulation targets, and permanent electrode implantation. After surgery, the patient significantly improved on both the Burke-Fahn-Marsden Dystonia Rating Scale and the Barry-Albright Dystonia Scale. The patient's response suggests that DBS can have potential benefit in H-ABC. TUBB4A mutations are associated with a variety of clinical phenotypes, and there is a lack of clearly identified targets for DBS, with this case being the second reported instance of DBS in this condition. The staged procedure with temporary depth electrode testing is recommended to identify optimal stimulation targets. The response seen in this patient implies that such a staged procedure may provide benefit in other conditions where DBS targets are currently unknown, including rare genetic or metabolic conditions associated with movement disorders.


Tracheal vascular response to hypertonic and hypotonic solutions.
One of the major roles of the upper respiratory mucosa is to humidify inspired air. This function requires the coordinated activity of respiratory epithelium and mucosal vasculature. It has been difficult to study this relationship in vivo. In the present study, we investigated the effects of osmolarity on the vessel diameter of mucosal vessels via a specially constructed chamber that allows direct visualization of the rat trachea microvasculature. With use of an anesthetized and instrumented rat preparation, the luminal surface of the dorsal wall of the trachea was superfused with physiological solutions at 37 degrees C. The osmolarities were varied by removing or adding NaCl or mannitol (200, 290, and 500 mosM). The mucosal vessels dilated when the airway surface was superfused with hypertonic solutions and constricted when superfused with hypotonic solution. The largest changes occurred in the arterioles (51 +/- 5.6 microns diam), which constricted by 10 +/- 2.18 microns (P = 0.0001) when exposed to a 200 mosM solution and dilated by 11 +/- 1.55 microns (P = 0.0001) when exposed to a 500 mosM NaCl-enriched solution. Smaller changes of similar pattern were seen in venules. The changes in vessel diameter were readily reversible upon replacement of the hypo- or hypertonic solutions by an isotonic solution. We conclude that increase or decrease of solution osmolarity on the epithelial surface of the trachea can regulate diameter of mucosal blood vessels.


Trigemino-cardiac reflex: occurrence of asystole during trans-sphenoidal adenomectomy: a case report.
The trigemino-cardiac reflex has been reported to occur during various craniofacial surgeries or procedures including manipulation of the trigeminal ganglion, tumor resection in the cerebellopontine angle, various facial reconstructions and trans-sphenoidal adenomectomy. Regarding risk factors during trans-sphenoidal adenomectomy, invasiveness closely related to the size of tumor and the degree of manipulation of cavernous sinus wall have been reported. We report the case of a 40-year-old female patient who had a relatively small-sized (< 10 mm) pituitary adenoma. Repetitive asystoles occurred during microscopic trans-sphenoidal operation of the wall of the cavernous sinus, which strongly suggests the importance of careful manipulation of the cavernous sinus wall. In addition to reporting this rare complication of trans-sphenoidal adenomectomy, we reviewed its clinical management by performing a literature search.


Primary structure, synthesis, and biological activity of rat endothelin, an endothelium-derived vasoconstrictor peptide.
Endothelin is a potent vasoconstrictor/pressor peptide, which we recently characterized from the conditioned culture medium of porcine aortic endothelial cells. We report here the cloning and partial sequencing of the rat endothelin gene. The nucleotide sequence predicted a 21-residue peptide similar to, but distinct from, porcine endothelin; 15 residues of rat endothelin were identical and 3 residues were substitutions by chemically similar amino acid residues to those in the porcine peptide. Synthetic rat endothelin was then prepared according to its deduced amino acid sequence. This synthetic peptide had (i) potent vasoconstrictor activity in the rat aortic strip and in perfused rat heart and (ii) a characteristically long-lasting in vivo pressor activity by intraaortic bolus injection in the conscious rat.


A Novel LSTM-Based Machine Learning Model for Predicting the Activity of Food Protein-Derived Antihypertensive Peptides.
Food protein-derived antihypertensive peptides are a representative type of bioactive peptides. Several models based on partial least squares regression have been constructed to delineate the relationship between the structure and activity of the peptides. Machine-learning-based models have been applied in broad areas, which also indicates their potential to be incorporated into the field of bioactive peptides. In this study, a long short-term memory (LSTM) algorithm-based deep learning model was constructed, which could predict the IC50 value of the peptide in inhibiting ACE activity. In addition to the test dataset, the model was also validated using randomly synthesized peptides. The LSTM-based model constructed in this study provides an efficient and simplified method for screening antihypertensive peptides from food proteins.


Porous starch and its application in drug delivery systems.
In recent years, starch has become a new potential biomaterial for pharmaceutical applications. This biopolymer has unique physicochemical and functional characteristics, as well as various advantages such as low price, relative ease of isolation in pure form from the plant source, non-toxicity, biodegradability, good biocompatibility, and interaction with living cells. Starch is currently used in pharmacy as a binder, disintegrating agent, film-forming material, raw material for production of microspheres and nanoparticles, and a component of drug delivery systems. Porous starch, which can be obtained with physical, chemical and enzymatic methods of modification, has a large specific surface area thanks to the presence of pores and channels. It has excellent adsorption capacity and can be used to enhance the dissolution rate of poorly soluble drugs or as shell material to improve the stability and water-solubility of compounds. As a component of drug delivery systems, porous starch has another advantage: it is biodegradable, so there is no need to remove it from the body after the release of the active agent.


Development of a QSAR model for predicting aqueous reaction rate constants of organic chemicals with hydroxyl radicals.
Reaction with hydroxyl radicals (˙OH) is an important removal pathway for organic pollutants in the aquatic environment. The aqueous reaction rate constant (kOH) is therefore an important parameter for fate assessment of aquatic pollutants. Since experimental determination fails to meet the requirement of being able to efficiently handle numerous organic chemicals at limited cost and within a relatively short period of time, in silico methods such as quantitative structure-activity relationship (QSAR) models are needed to predict kOH. In this study, a QSAR model with a larger and wider applicability domain as compared with existing models was developed. Following the guidelines for the development and validation of QSAR models proposed by the Organization for Economic Co-operation and Development (OECD), the model shows satisfactory performance. The applicability domain of the model has been extended and contained chemicals that have rarely been covered in most previous studies. The chemicals covered in the current model contain functional groups including [double bond splayed left]C[double bond, length as m-dash]C[double bond splayed right], -C[triple bond, length as m-dash]C-, -C6H5, -OH, -CHO, -O-, [double bond splayed left]C[double bond, length as m-dash]O, -C[double bond, length as m-dash]O(O)-, -COOH, -C[triple bond, length as m-dash]N, [double bond splayed left]N-, -NH2, -NH-C(O)-, -NO2, -N[double bond, length as m-dash]C-N[double bond splayed right], [double bond splayed left]N-N[double bond splayed right], -N[double bond, length as m-dash]N-, -S-, -S-S-, -SH, -SO3, -SO4, -PO4, and -X (F, Cl, Br, and I).


Naïve Pine Terpene Response to the Mountain Pine Beetle (Dendroctonus ponderosae) through the Seasons.
Insect herbivores must contend with constitutive and induced plant defenses. The mountain pine beetle, Dendroctonus ponderosae Hopkins (Coleoptera: Curculionidae, Scolytinae) has expanded its range east of the Rocky Mountains into the western boreal forest and is encountering evolutionarily naïve lodgepole pines (Pinus contorta) and jack pines (Pinus banksiana). Pinus contorta and P. banksiana in the expanded range have different constitutive and induced defenses in response to wounding and inoculation with fungal associates of D. ponderosae. In the historic range, previous studies have examined phloem terpene content prior to and just after D. ponderosae mass attack, but the terpene profile of attacked trees post-overwintering is unknown. We examined the response of mature P. contorta and P. banksiana trees to experimentally-applied mass attack by D. ponderosae and quantified phloem terpenes at three time points, pre-attack, post-attack (same season), and the following spring, post-overwintering. Phloem content of total terpenes as well as many individual terpenes increased after D. ponderosae attack but were only significantly higher than pre-attack levels at the post-overwintering time point in both P. contorta and P. banksiana. The absence of a significant increase in phloem terpenes in the month following attack in naïve pines is a potential cause for increased D. ponderosae offspring production reported in naïve P. contorta. Beetle attack density did not influence the phloem terpene profiles of either species and there was no significant interaction between attack density and sampling time on terpene content. High phloem terpenes in trees that are attacked at low densities could prime these trees for defense against attacks in the following season but it could also make these trees more apparent to early-foraging beetles and facilitate efficient mass attack at low D. ponderosae population densities in the expanded range.


Construction of an autophagy-related eleven long noncoding RNA signature to predict the outcomes, immune cell infiltration, and immunotherapy response in patients with gastric cancer.
Long noncoding RNAs (LncRNAs) may be involved in the occurrence, development, and drug resistance of gastric cancer (GC) by regulating autophagy. This study aims to establish an autophagy-related LncRNA (ARL) signature (ARLSig) and explore its immunogenomic implications in patients with GC. The RNA sequencing and clinical data of patients with GC from The Cancer Genome Atlas database, and autophagy genes from the Human Autophagy Database were extracted. The co-expression and Cox regression analyses were performed to establish a prognostic ARLSig. Further, the differences in clinicopathology, immune microenvironment, immune function, and response to immunotherapy between the risk groups were explored by several algorithms. A prognostic risk model consisting of 11 ARLs was constructed. The clinical correlation analysis between the ARLSig and clinicopathological factors indicated that the ARLSig was correlated with the comprehensive, T, and N stages (all P<0.05). Further, a nomogram including the ARLSig and clinical factors suggested it had a powerful predictive value for survival, with a higher prediction efficiency for 1-, 3-, and 5-year survival than other clinicopathological factors. Finally, the immune-related analysis between the two risk groups showed that the high-risk group had significantly higher infiltration proportions of natural killer cells resting, monocytes, M2 macrophages, and dendritic cells resting, as well as higher expression of 25 immune checkpoint genes. In addition, the immunotherapy response prediction by the tracking of indels by decomposition algorithm showed the low-risk group was more sensitive to immune checkpoint inhibitor therapy. The ARLSig consisting of 11 ARLs in GC showed highly efficient predictive value for survival of patients with GC and might provide novel targets for their individualized immunotherapy.


Left ventricular long axis function assessed during cine-cardiovascular magnetic resonance is an independent predictor of adverse cardiac events.
Left ventricular pump function requires a complex interplay involving myocardial fibers orientated in the longitudinal, oblique and circumferential directions. Long axis dysfunction appears to be an early marker for a number of pathological states. We hypothesized that mitral annular plane systolic excursion (MAPSE) measured during cine-cardiovascular magnetic resonance (CMR) reflects changes in long axis function and may be an early marker for adverse cardiovascular outcomes. The aims of this study were therefore: 1) To assess the feasibility and reproducibility of MAPSE measurements during routine cine-CMR; and 2) To assess whether MAPSE, as a surrogate for long axis function, is a predictor of major adverse cardiovascular events (MACE). Four hundred consecutive patients undergoing CMR were prospectively enrolled. MAPSE was measured in the 4-chamber cine view. Patients were prospectively followed for major adverse cardiac events (MACE) - death, non-fatal myocardial infarction, hospitalization for heart failure or unstable angina, and late revascularization. Cox proportional hazards regression modeling was used to identify factors independently associated with MACE. Net reclassification improvement (NRI) was calculated to assess whether addition of MAPSE resulted in improved risk reclassification of MACE. Seventy-two MACE occurred during a median follow-up of 14.5 months. By Kaplan-Meier analysis, patients with lateral MAPSE <1.11 cm (median) experienced significantly higher incidence of MACE than patients with a MAPSE ≥1.11 cm (p = 0.027). After adjustment for established clinical risk factors which were univariate predictors (age, diabetes, hypertension, NYHA class, LV mass), lateral MAPSE remained a significant independent predictor of MACE (HR = 4.384 per cm decrease or 1.344 per 2 mm decrease; p = 0.020). Incorporation of lateral MAPSE into this risk model resulted in a net reclassification improvement (NRI) of 0.18 (p = 0.006). Reduced long axis function assessed with lateral MAPSE during cine-CMR is an independent predictor of MACE.


Peripheral insulin resistance predicts liver damage in nondiabetic subjects with nonalcoholic fatty liver disease.
Surrogate indexes of insulin resistance and insulin sensitivity are widely used in nonalcoholic fatty liver disease (NAFLD), although they have never been validated in this population. We aimed to validate the available indexes in NAFLD subjects and to test their ability to predict liver damage also in comparison with the NAFLD fibrosis score. Surrogate indexes were validated by the tracer technique (6,6-D2 -glucose and U-(13) C-glucose) in the basal state and during an oral glucose tolerance test. The best-performing indexes were used in an independent cohort of 145 nondiabetic NAFLD subjects to identify liver damage (fibrosis and nonalcoholic steatohepatitis). In the validation NAFLD cohort, homeostasis model assessment of insulin resistance, insulin to glucose ratio, and insulin sensitivity index Stumvoll had the best association with hepatic insulin resistance, while peripheral insulin sensitivity was most significantly related to oral glucose insulin sensitivity index (OGIS), insulin sensitivity index Stumvoll, and metabolic clearance rate estimation without demographic parameters. In the independent cohort, only oral glucose tolerance test-derived indexes were associated with liver damage and OGIS was the best predictor of significant (≥F2) fibrosis (odds ratio = 0.76, 95% confidence interval 0.61-0.96, P = 0.0233) and of nonalcoholic steatohepatitis (odds ratio = 0.75, 95% confidence interval 0.63-0.90, P = 0.0021). Both OGIS and NAFLD fibrosis score identified advanced (F3/F4) fibrosis, but OGIS predicted it better than NAFLD fibrosis score (odds ratio = 0.57, 95% confidence interval 0.45-0.72, P < 0.001) and was also able to discriminate F2 from F3/F4 (P < 0.003). OGIS is associated with peripheral insulin sensitivity in NAFLD and inversely associated with an increased risk of significant/advanced liver damage in nondiabetic subjects with NAFLD.


[Single photon emission computed tomography findings in a case of alternating hemiplegia of childhood in relation to migraine].
Single photon emission computed tomography (SPECT) was performed 3 times during attacks and performed 4 times during postictal periods on a case of alternating hemiplegia of childhood. Hyperperfusion of the corresponding hemisphere to hemiparesis was suggested by asymmetric increase RI uptake during the ictal scans, whereas interictal scans showed symmetric topography of cerebral blood flow. Manifestations except hemiplegia included loss of consciousness and vomiting. These manifestations and ictal SPECT findings are the same evidence as hemiplegic migraine. It suggests that these two disorders have a similar pathophysiology. There are two interesting findings regarding this patient. The first finding is magnetic resonance imaging showed progressive cerebellar atrophy. Second finding is interictal SPECT showed a progressive decrease of cerebral perfusion, especially in cerebellar hemispheres. These two findings suggest alternating hemiplegia of childhood may be a chronic progressive disorder.


Cloning of a novel tyrosinase-encoding gene (melB) from Aspergillus oryzae and its overexpression in solid-state culture (Rice Koji).
We have cloned a novel tyrosinase-encoding gene (melB) specifically expressed in solid-state culture of Aspergillus oryzae. A tyrosinase-encoding gene (melO) from A. oryzae was already cloned and the protein structures of its catalytic and copper binding domains were investigated. However, our recent results revealed that the melO gene was highly expressed in submerged culture but not in solid-state culture. Because tyrosinase activity was also detected in solid-state culture, we assumed that another tyrosinase gene other than melO is expressed in solid-state culture. Another tyrosinase gene was screened using the expressed sequence tag (EST) library. One redundant cDNA clone homologous with the tyrosinase gene was found in the collection of wheat bran culture. Northern blot analysis revealed that the gene corresponding to the cDNA clone was specifically expressed in solid-state culture (koji making), but not in submerged culture. Molecular cloning showed that the gene carried six exons interrupted by five introns and had an open reading frame encoding 616 amino acid residues. This gene was designated as melB. The deduced amino acid sequence of the gene had weak homology (24%-33%) with MelO and other fungal tyrosinases but the sequences of the copper binding domains were highly conserved. When the melB gene was expressed under the control of the glaB promoter in solid-state culture, tyrosinase activity was markedly enhanced and the culture mass was browned with the melanization by MelB tyrosinase. These results indicated that the melB gene encodes a novel tyrosinase associated with melanization in solid-state culture.


Interleukin-17 in Chronic Inflammatory Neurological Diseases.
A critical role for IL-17, a cytokine produced by T helper 17 (Th17) cells, has been indicated in the pathogenesis of chronic inflammatory and autoimmune diseases. A positive effect of blockade of IL-17 secreted by autoreactive T cells has been shown in various inflammatory diseases. Several cytokines, whose production is affected by environmental factors, control Th17 differentiation and its maintenance in tissues during chronic inflammation. The roles of IL-17 in the pathogenesis of chronic neuroinflammatory conditions, multiple sclerosis (MS), experimental autoimmune encephalomyelitis (EAE), Alzheimer's disease, and ischemic brain injury are reviewed here. The role of environmental stimuli in Th17 differentiation is also summarized, highlighting the role of viral infection in the regulation of pathogenic T helper cells in EAE.


Tryptic Peptides Bearing C-Terminal Dimethyllysine Need to Be Considered during the Analysis of Lysine Dimethylation in Proteomic Study.
Lysine methylation plays important roles in structural and functional regulation of chromatin. Although trypsin is the most widely used protease in mass spectrometry-based proteomic analysis for lysine methylation substrates, the proteolytic activity of trypsin on dimethylated lysine residues remains an arguable issue. In this study, we tested the ability of trypsin to cleave dimethylated lysine residues in synthetic peptides, purified albumin, and whole cell lysate, and found that the C-terminal of dimethylated lysine residue could be cleaved in a protein sequence-dependent manner. Kinetic studies revealed that the optimal digestion time and enzyme-to-substrate ratio for the cleavage of dimethylated lysine by trypsin was around 16 h and 1:50, respectively. We further showed the tryptic C-terminal lysine-dimethylated (C-Kme2) peptides could contribute to a significant portion of substrate identification in the proteomic study, which utilizes the chemical dimethylation labeling approach. More than 120 tryptic C-Kme2 peptides (7% of total peptides identified) were identified in chemically lysine-dimethyl-labeled HeLa whole cell lysate by a single-shot nanoflow high performance liquid chromatography with tandem mass spectrometry (nano-HPLC-MS/MS) analysis. Moreover, in an assay for substrate identification of protease Glu-C using stable isotope dimethyl labeling approach, our data showed the tryptic C-Kme2 peptides accounted for more than 13% of total tryptic peptides. Additionally, our in vivo methylome profiling data revealed some C-Kme2 peptides, which is of importance to identification and quantification of biologically relevant protein and lysine-methylated site. Therefore, we reason that the tryptic peptides bearing C-terminal dimethylated lysine need to be considered in the mass spectrometric analysis of lysine dimethylation.


Artificial Neural Network Approach in Laboratory Test Reporting: Learning Algorithms.
In the field of laboratory medicine, minimizing errors and establishing standardization is only possible by predefined processes. The aim of this study was to build an experimental decision algorithm model open to improvement that would efficiently and rapidly evaluate the results of biochemical tests with critical values by evaluating multiple factors concurrently. The experimental model was built by Weka software (Weka, Waikato, New Zealand) based on the artificial neural network method. Data were received from Dokuz Eylül University Central Laboratory. "Training sets" were developed for our experimental model to teach the evaluation criteria. After training the system, "test sets" developed for different conditions were used to statistically assess the validity of the model. After developing the decision algorithm with three iterations of training, no result was verified that was refused by the laboratory specialist. The sensitivity of the model was 91% and specificity was 100%. The estimated κ score was 0.950. This is the first study based on an artificial neural network to build an experimental assessment and decision algorithm model. By integrating our trained algorithm model into a laboratory information system, it may be possible to reduce employees' workload without compromising patient safety.


Improving geriatrics training in internal medicine residency programs: best practices and sustainable solutions.
National surveys indicate a need for additional training in geriatrics during internal medicine residencies. This paper describes 1) "best practices" for integrating geriatrics education into internal medicine residency programs, 2) barriers to implementation of these practices, and 3) possible ways to improve geriatrics training for internal medicine residents. These best practices were determined by a systematic review of the literature and through interviews with leaders of 26 residency and geriatrics programs concerned with geriatrics training for residents. The most successful programs have clinical experiences with 3 key elements: model geriatric care in 1 or more settings (for example, in the hospital or in ambulatory practice), patient care across sites or transitions of care, and interdisciplinary teamwork. Barriers include attitudes, few faculty, need for relationships with nontraditional training sites, and lack of funding. Local solutions include engaging the internal medicine program director to accomplish a mutual goal--for example, by creating a model geriatrics training experience in which residents demonstrate their skill in a new Accreditation Council of Graduate Medical Education competency (such as systems-based practice). National solutions include reaching consensus on the competencies in geriatrics that should be achieved by board-eligible internists. This may mean increasing the number of questions that test geriatrics competency in the certifying and in-training examinations, increasing numbers of faculty members able to teach and model geriatric care, developing "effective medical resident teaching" courses for nonphysician faculty, and lobbying for improved systems of care.


Low genetic variability in the highly endangered mediterranean monk seal.
Genetic variability is an important component in the ability of populations to adapt in the face of environmental change. Here we report the first description of nuclear genetic variability in the only remaining sizable colony of the Mediterranean monk seal (Monachus monachus), located at Cap Blanc (Western Sahara, Mauritania), whose estimated size during the study period (1994-May 1997) was about 320 individuals. We tested 42 microsatellite loci isolated from five pinniped species in a sample of 52 pups. Three loci failed to give any product, and of the remaining 39, only 15 were polymorphic, with a maximum of 3 alleles detected. Three loci appeared to be X-linked. No departures from Hardy-Weinberg equilibrium were detected and no genetic structure was found between the two nursing caves currently occupied by the seals. Several analytical methods show that, as a consequence of a severe bottleneck, the population has suffered a decrease in genetic variability over the last few centuries.


Exchange protein directly activated by cAMP (epac): a multidomain cAMP mediator in the regulation of diverse biological functions.
Since the discovery nearly 60 years ago, cAMP is envisioned as one of the most universal and versatile second messengers. The tremendous feature of cAMP to tightly control highly diverse physiologic processes, including calcium homeostasis, metabolism, secretion, muscle contraction, cell fate, and gene transcription, is reflected by the award of five Nobel prizes. The discovery of Epac (exchange protein directly activated by cAMP) has ignited a new surge of cAMP-related research and has depicted novel cAMP properties independent of protein kinase A and cyclic nucleotide-gated channels. The multidomain architecture of Epac determines its activity state and allows cell-type specific protein-protein and protein-lipid interactions that control fine-tuning of pivotal biologic responses through the "old" second messenger cAMP. Compartmentalization of cAMP in space and time, maintained by A-kinase anchoring proteins, phosphodiesterases, and β-arrestins, contributes to the Epac signalosome of small GTPases, phospholipases, mitogen- and lipid-activated kinases, and transcription factors. These novel cAMP sensors seem to implement certain unexpected signaling properties of cAMP and thereby to permit delicate adaptations of biologic responses. Agonists and antagonists selective for Epac are developed and will support further studies on the biologic net outcome of the activation of Epac. This will increase our current knowledge on the pathophysiology of devastating diseases, such as diabetes, cognitive impairment, renal and heart failure, (pulmonary) hypertension, asthma, and chronic obstructive pulmonary disease. Further insights into the cAMP dynamics executed by the Epac signalosome will help to optimize the pharmacological treatment of these diseases.


Positional changes of a pluripotency marker gene during structural reorganization of fibroblast nuclei in cloned early bovine embryos.
Cloned bovine preimplantation embryos were generated by somatic cell nuclear transfer (SCNT) of bovine fetal fibroblasts with a silent copy of the pluripotency reporter gene GOF, integrated at a single site of a chromosome 13. GOF combines the regulatory Oct4/Pou5f1 sequence with the coding sequence for EGFP. EGFP expression served as a marker for pluripotency gene activation and was consistently detected in preimplantation embryos with 9 and more cells. Three-dimensional radial nuclear positions of GOF, its carrier chromosome territory and non-carrier homolog were measured in nuclei of fibroblasts, and of day 2 and day 4 embryos, carrying 2 to 9 and 15 to 22 cells, respectively. We tested, whether transcriptional activation was correlated with repositioning of GOF toward the nuclear interior either with a corresponding movement of its carrier chromosome territory 13 or via the formation of a giant chromatin loop. A significant shift of GOF away from the nuclear periphery was observed in day 2 embryos together with both carrier and non-carrier chromosome territories. At day 4, GOF, its carrier chromosome territory 13 and the non-carrier homolog had moved back toward the nuclear periphery. Similar movements of both chromosome territories ruled out a specific GOF effect. Pluripotency gene activation was preceded by a transient, radial shift of GOF toward the nuclear interior. The persistent co-localization of GOF with its carrier chromosome territory rules out the formation of a giant chromatin loop during GOF activation.


Specificity of Quantitative Functional Brain Mapping with Arterial Spin-Labeling for Preoperative Assessment.
Arterial spin-labeling is a noninvasive MR imaging technique allowing direct and quantitative measurement of brain perfusion. Arterial spin-labeling is well-established in clinics for investigating the overall cerebral perfusion, but it is still occasionally employed during tasks. The typical contrast for functional MR imaging is blood oxygen level-dependent (BOLD) imaging, whose specificity could be biased in neurologic patients due to altered neurovascular coupling. This work aimed to validate the use of functional ASL as a noninvasive tool for presurgical functional brain mapping. This is achieved by comparing the spatial accuracy of functional ASL with transcranial magnetic stimulation as the criterion standard. Twenty-eight healthy participants executed a motor task and received a somatosensory stimulation, while BOLD imaging and arterial spin-labeling were acquired simultaneously. Transcranial magnetic stimulation was subsequently used to define hand somatotopy. Functional ASL was found more adjacent to transcranial magnetic stimulation than BOLD imaging, with a significant shift along the inferior-to-superior direction. With respect to BOLD imaging, functional ASL was localized significantly more laterally, anteriorly, and inferiorly during motor tasks and pneumatic stimulation. Our results confirm the specificity of functional ASL in targeting the regional neuronal excitability. Functional ASL could be considered as a valid supplementary technique to BOLD imaging for presurgical mapping when spatial accuracy is crucial for delineating eloquent cortex.


Mild hypothermia protects auditory function during cochlear implant surgery.
Loss of auditory function after cochlear implant (CI) electrode insertion occurs in two stages in the laboratory rat. An immediate loss is followed by a progressive loss over 7 days. Similar stages of acute and progressive neuronal loss occur after trauma in the central nervous system where hypothermia has been shown to have a protective effect. We hypothesize that hypothermia has a similar protective effect against loss of auditory function caused by CI electrode insertion trauma. Thirty rats underwent surgery in one cochlea; the contralateral ear was an unoperated control. In the normothermia group, CI electrode insertion trauma was generated with rectal temperature maintained at 37 degrees C throughout the experiment. In the mild hypothermia group, electrode trauma was generated with rectal temperature lowered to 34 degrees C. In the surgical control group, mock surgery was performed at 37 degrees C. Multiple frequency auditory brainstem response (ABR) and distortion product otoacoustic emission (DPOAE) testing of all ears was performed before surgery, immediately afterward, and on postoperative days 3, 5, and 7. Both ABR and DPOAE testing demonstrated partial loss of auditory function after electrode insertion trauma. However, the hypothermia group had significantly less functional loss in the immediate stage and no significant loss in the progressive stage. Mild hypothermia protects auditory function during CI electrode insertion.


Leptin regulates disc cartilage endplate degeneration and ossification through activation of the MAPK-ERK signalling pathway in vivo and in vitro.
Recent findings demonstrate that leptin plays a significant role in chondrocyte and osteoblast differentiation. However, the mechanisms by which leptin acts on cartilage endplate (CEP) cells to give rise to calcification are still unclear. The aim of this study was to evaluate the effects of leptin that induced mineralization of CEP cells in vitro and in vivo. We constructed a rat model of lumbar disc degeneration and determined that leptin was highly expressed in the presence of CEP calcification. Rat CEP cells treated with or without leptin were used for in vitro analysis using RT-PCR and Western blotting to examine the expression of osteocalcin (OCN) and runt-related transcription factor 2 (Runx2). Both OCN and Runx2 expression levels were significantly increased in a dose- and time-dependent manner. Leptin activated ERK1/2 and STAT3 phosphorylation in a time-dependent manner. Inhibition of phosphorylated ERK1/2 using targeted siRNA suppressed leptin-induced OCN and Runx2 expression and blocked the formation of mineralized nodules in CEP cells. We further demonstrated that exogenous leptin induced matrix mineralization of CEP cells in vivo. We suggest that leptin promotes the osteoblastic differentiation of CEP cells via the MAPK/ERK signal transduction pathway and may be used to investigate the mechanisms of disc degeneration.


Ultrasonographic adrenal gland measurements in healthy Yorkshire Terriers and Labrador Retrievers.
An upper threshold of 7.4 mm for maximal adrenal gland diameter is commonly used to detect pituitary-dependent hyperadrenocorticism ultrasonographically in dogs. There is a substantial overlap between adrenal gland diameter of healthy dogs and of those with pituitary-dependent hyperadrenocorticism. The aim of this study is to determine the measurements of both adrenal glands, in particular, of the height at the caudal glandular pole in a longitudinal plane, in the Labrador retriever and Yorkshire terrier, two breeds widely represented in the population suspected of hyperadrenocorticism. Seventeen Labrador retrievers and 24 Yorkshire terriers considered healthy were included in the study. Adrenal gland measurements were taken on static images and comprised in measurements of the length in a longitudinal plane (L), of the height at the cranial (CrHLG) and caudal pole (CdHLG) in a longitudinal plane and in a transverse plane (CrHTR and CdHTR, respectively), and of the width at the cranial and caudal poles in a transverse plane (CrWTR and CdWTR, respectively). This study established new upper thresholds for the left and right height at the caudal pole measured in a longitudinal plane: 7.9 mm (left) and 9.5 mm (right) for the Labrador retrievers and 5.4 mm (left) and 6.7 mm (right) for the Yorkshire terriers. All the measurements were significantly different between the two breeds. There was a significant relationship between CdHTR and CdHLG, and the age of the dogs for both breeds.


The macula in ocular toxoplasmosis.
Of 60 attacks of ocular toxoplasmosis, in 37 (62%) no apparent macular problems were demonstrated on inspection, in seven (12%) active retinitis was evident within 5 degrees of the umbo, and in seven (12%) some evidence was shown of mild macular edema; severe edema, such as cystoid macular edema, was uncommon.


Doppler Audio Signal Analysis as an Additional Tool in Evaluation of Umbilical Artery Circulation.
Purpose To investigate the predictive capacity of a new method for sound spectrum analysis of Doppler signals recorded from the umbilical artery in high-risk pregnancies. Material and Methods The retrospective study comprised 127 pregnant women with various pregnancy complications between 23 and 39 gestational weeks. Umbilical artery blood flow velocity waveforms were recorded with Doppler ultrasound and characterized by pulsatility index (PI) and blood flow class (BFC). Doppler audio signals were stored on a digital video recorder and the sound frequency at the energy level 15 dB below its peak (MAXpeak-15 dB) was estimated off-line. The prediction of probability for composite adverse pregnancy outcome (operative delivery for fetal distress, admission to neonatal intensive care unit, perinatal death) was evaluated using the area under the curve (AUC) of the receiver operating characteristics (ROC) curve. Results With increasing umbilical artery BFC, the MAXpeak-15 dB frequencies decreased (p < 0.0001) and the PI increased (p < 0.0001). The ROC AUCs for adverse outcome for MAXpeak-15 dB and for PI were 0.842 and 0.836 (p = 0.88), respectively. For the combination of MAXpeak-15 dB and PI, the corresponding AUC was 0.894, significantly higher than that of PI (p < 0.03) and of MAXpeak-15 dB (p < 0.05). Conclusion Umbilical artery Doppler sound spectrum analysis might be a useful supplement to PI in the clinical evaluation of fetoplacental circulation.


Adaptation of vasomotor function of human coronary arterioles to the simultaneous presence of obesity and hypertension.
We hypothesized that simultaneous presence of obesity and hypertension activates adaptive vascular mechanisms affecting dilations of human coronary arterioles. Agonist-induced dilations were assessed in isolated pressurized coronary arterioles from patients (n=38) who underwent cardiac surgery. Among normotensives we found that dilations to bradykinin (BK) and the NO-donor, sodium-nitroprusside (SNP) were reduced in obese subjects (BK, 10(-7) mol/L, lean: 90+/-4%, obese: 64+/-7%; SNP, 10(-6) mol/L, lean: 89+/-7%, obese: 76+/-5%). However, among hypertensives, both BK- and SNP-induced dilations were significantly enhanced in obese patients, when compared with lean individuals (BK, lean: 71+/-7%, obese: 85+/-3%; SNP, lean: 60+/-6%, obese: 83+/-2%). Correspondingly, in hypertensive patients, but not in those of normotensives, a positive correlation was found between body mass index (BMI) and BK-induced (P=0.036, r=0.46), and also SNP-evoked (P=0.031, r=0.44) coronary dilations. Moreover, in additional 55 hypertensive patients flow-mediated (FMD) and nitroglycerin (NTG)-induced dilations of the brachial artery were assessed. In obese hypertensive individuals, FMD- and NTG-induced dilations were greater (FMD: 6.2+/-0.7%, NTG: 17.2+/-0.9%), than in lean hypertensive patients (FMD: 3.7+/-0.6%, NTG: 13.6+/-1.1%). Correspondingly, FMD- and NTG-induced dilations were positively correlated with BMI (P=0.020, r=0.31 and P=0.033, r=0.29, respectively). These findings are the first to suggest that obesity may lead to activation of adaptive vascular mechanisms to enhance the dilator function of coronary and peripheral arterial vessels in hypertensive patients.


Serum beta-N-acetylglucosaminidase in patients with symptomatic porphyria.
Studies in 30 male patients with symptomatic porphyria reveal an increased activity in the serum of the lysosomal enzyme beta-N-acetylglucosaminidase (EC 220.127.116.11) as compared with a control group of 33 healthy males. Enzymatic activity is higher in the active stage of the disease (with fresh skin lesions) than in patients found in remission. A high degree of correlation has been detected between the activity of beta-N-acetylglucosaminidase in the serum and uroporphyrin in the urine, and the total plasma porphyrins. The increased activity of this enzyme in the serum indicates that the lysosomes participate in the pathogenesis of this disorder. Further studies are warranted for the complete elucidation of the clinical significance of this parameter.


[Simple analysis of fatty acid in human milk by HPLC].
The conventional analysis of fatty acid requires such complicated operations as extraction of total lipid, evaporation of organic layer, alkaline hydrolysis, washing with n-hexane, re-extraction of fatty acid and washing with water, and therefore, is time-consuming. Herein, we report a simplified method dispensing with the steps of the extraction of total lipids, the evaporation of organic layer and the washing with n-hexane after hydrolysis. This method showed satisfactory results on the linearity of standard curves and their productibility, and a correlation was obtained between those obtained by the traditional method and by our simple method in human milk. We employed this method for the analysis of human milk secreted between the 3rd day and the 42nd day after delivery. The concentration of total fatty acid in human milk secreted after lactation showed a higher value than that before it, while the constituent ratio of fatty acid did not show such a great difference.


Resolution-by-proxy: a simple measure for assessing and comparing the overall quality of NMR protein structures.
In protein X-ray crystallography, resolution is often used as a good indicator of structural quality. Diffraction resolution of protein crystals correlates well with the number of X-ray observables that are used in structure generation and, therefore, with protein coordinate errors. In protein NMR, there is no parameter identical to X-ray resolution. Instead, resolution is often used as a synonym of NMR model quality. Resolution of NMR structures is often deduced from ensemble precision, torsion angle normality and number of distance restraints per residue. The lack of common techniques to assess the resolution of X-ray and NMR structures complicates the comparison of structures solved by these two methods. This problem is sometimes approached by calculating "equivalent resolution" from structure quality metrics. However, existing protocols do not offer a comprehensive assessment of protein structure as they calculate equivalent resolution from a relatively small number (<5) of protein parameters. Here, we report a development of a protocol that calculates equivalent resolution from 25 measurable protein features. This new method offers better performance (correlation coefficient of 0.92, mean absolute error of 0.28 Å) than existing predictors of equivalent resolution. Because the method uses coordinate data as a proxy for X-ray diffraction data, we call this measure "Resolution-by-Proxy" or ResProx. We demonstrate that ResProx can be used to identify under-restrained, poorly refined or inaccurate NMR structures, and can discover structural defects that the other equivalent resolution methods cannot detect. The ResProx web server is available at http://www.resprox.ca.


Monitoring communication with patients: analyzing judgments of satisfaction (JOS).
Medical students struggle to put into practice communication skills learned in medical school. In order to improve our instructional designs, better insight into the cause of this lack of transfer is foundational. We therefore explored students' cognitions by soliciting self-evaluations of their history-taking skills, coined 'judgments of satisfaction (JOSs)'. Our cognitive-psychological approach was guided by Koriat's cue-utilization framework (J Exp Psychol Gen 126:349-370. doi: 10.1037/0096-34184.108.40.2069 , 1997) which rests on the assumption that internal and external cues inform learners' metacognitive judgments, which, in turn, steer their actions. Judgments based on unsuitable cues will cause ineffective behavior. Consequently, students are unable to adequately master these skills or properly apply them in similar situations. For the analysis, we had 524 medical undergraduates select scenes they were satisfied or dissatisfied with from their video-recorded simulated-patient encounters and explain why. Twenty transcripts were sampled for directed content analysis. We found that approximately one-third of students' judgments focused on content (JOS-type-a); about half on the quality of the communication skills (JOS-type-b); and about ten percent targeted the appropriateness of the skills harnessed (JOS-type-c). This lack of reflection on appropriateness may explain why students experience problems adapting to new situations. It was primarily high-performance students who formed type-c judgments; poor performers tended to give type-a and type-b judgments. Future research would benefit from the use of our modified version of Koriat's framework in order to further explore how high and poor performing medical students differ in the way they form JOSs during communications skills training.


Central noradrenergic neurons: identification, distribution and synaptic interactions with axons containing morphine-like peptides.
Perikarya and processes of noradrenergic neurons have been identified in brain by fluorescence histochemistry, radioautography, and immunocytochemistry. The neuronal perikarya are localized in discrete nuclei within the brain stem. Many of these nuclei such as the locus coeruleus send efferent projections to virtually every portion of the central nervous system. Electron-microscopic immunocytochemical studies suggest that the functioning of many of the noradrenergic neurons may be modulated by direct synaptic interactions with neurons containing the opiate-like peptides, methionine (Met5) and leucine (Leu5) enkephalin.


Kinetics and mechanism of degradation of dichlorvos in aqueous solutions containing reduced sulfur species.
Reactions of dichlorvos with five reduced sulfur species (hydrogen sulfide, bisulfide, thiosulfate, thiophenol, and thiophenolate) were examined in well-defined anoxic aqueous solutions to investigate their role in its degradation. Reactions were monitored at varying concentrations of reduced sulfur species over pH range to obtain the second-order reaction rate constants. Experiments at 25 degrees C demonstrated that degradation of dichlorvos promoted by bisufide, thiosulfate, and thiophenolate were of much greater importance than hydrolysis under the experimental conditions in our study. In contrast, hydrogen sulfide and thiophenol were not effective in the degradation of dichlorvos. The activation parameters of the reaction of dichlorvos with bisulfide, thiosulfate, and thiophenolate were also determined from the measured second-order rate constants over a temperature range of 12-50 degrees C. The relative reactivity of the reduced sulfur species decreases in the following order: PhS- > HS- approximately equal to S2O3(2-). When the second-order rate constants at 25 degrees C are multiplied by the environmentally relevant concentration of the reduced sulfur species, predicted half-lives of dichlorvos ranged from hours to days. The results indicated that reduced sulfur species could play a very important role in the chemical fate of dichlorvos in coastal marine environments.


Weil syndrome coincident with upper gastrointestinal bleeding.
A 48 year old male was referred to our center due to a gastrointestinal bleeding with melena secondary to a Forrest IIb gastric ulcer treated endoscopically. Physical examination revealed bilateral conjunctival suffusion, bradypsychia, and asterixis. Epidemiological history included a trip to Dominican Republic two weeks before, presenting later a flu-like syndrome. He had no history of NSAID use. Laboratory tests showed a normocytic anemia, leukocytosis with neutrophilia, acute renal failure, severe hyponatremia, a predominant direct hyperbilirubinemia, hyperamylasemia, and mild coagulopathy (Table 1). An abdominal ultrasound was performed, with no pathological findings, and a chest-abdominal computed tomography (CT), bilateral diffuse ground glass pulmonary opacities and pleural effusion, mild hepatomegaly, and peritoneal and gastrohepatic ligament lymphadenopathy, with no signs of acute pancreatitis. A second look upper endoscopy revealed a Forrest III gastric ulcer. Gastric biopsies results ruled out malignancy and Helicobacter pylori infection. Due to his recent travel history combined with his characteristic signs and symptoms a clinical diagnosis of leptospirosis was made and empirical antibiotic therapy with meropenem was started. The serology for Leptospira was positive (IgG 1/1600) and antibiotic therapy was de-escalated to ceftriaxone with clinical and analytical remission on day five of his hospital stay with complete radiological resolution at 6 months.


SP1-induced overexpression of LINC00520 facilitates non-small cell lung cancer progression through miR-577/CCNE2 pathway and predicts poor prognosis.
Long noncoding RNAs (lncRNAs) have gained much attention in the past few years. Long intergenic non-protein coding RNA 520 (LINC00520) was one of the newly discovered lncRNA which has been demonstrated to be dysregulated in several cancers. So far, the function and mechanism of LINC00520 in non-small cell lung cancer (NSCLC) are unclear. In this paper, our group first showed that LINC00520 level was elevated in non-small cell lung cancer (NSCLC) tissue and cells. In addition, SP1 could bind directly to the promoter region of LINC00520 and thus promote its transcription. Increased LINC00520 was distinctly correlated with advanced tumor stage and shorter survival time in NSCLC patients. Further functional investigations provided evidences that forced down regulation of LINC00520 inhibited NSCLC cell proliferation, invasion, metastasis and EMT, while contributing to cells apoptosis. Mechanistically, we found that LINC00520 serving as a competing endogenous RNA to be involved in the modulation of miR-577 expressions, and thus affected the expression of CCNE2 which was a target gene of miR-577. Moreover, in NSCLC cells with si-LINC00520, up regulation of CCNE2 led to an increase of cell growth and invasion. Taken together, LINC00520 displayed its tumor-promotive roles through modulating the miR-577/CCNE2 axis, highlighting a potential therapeutic strategy for NSCLC patients.