'Proximity frequencies' a new parameter to evaluate the profile of GABAAR modulators.
We reported the synthesis of new 8-methoxypyrazolo[1,5-a]quinazolines bearing an amide fragment at the 3-position. The final compounds, as aromatic (2a-i) and 4,5-dihydro derivatives (3a-i), have been evaluated in vitrofor their ability to modulate the chlorine current on recombinant GABAA receptors of the α1β2γ2L type (expressed in frog oocytes of the Xenopus laevis species). From electrophysiological test two groups of compounds emerged: positive modulators agonist (2e, h, i and 3e, h) and null modulators antagonist (2a, b, d, f, g and 3a-d, f, g) of GABAA subtype receptor. Using a set of compounds (new derivatives, known products and GABAA subtype receptor ligands from our library) we identify the amino acids at the α+/γ- interface, which could be involved in the agonist or antagonist profile, using the 'Proximity Frequencies', namely the frequencies with which a ligand intercepts two or more binding-site amino acids during the molecular dynamic simulation. The linear discriminant analysis (LDA) evidences that the combination of amino acids αVAL203- γTHR142 and αTYR 160- γTYR 58 allowed to collocate 70.6% of agonists and 72.7% of antagonists in their respective class.


The effects of the specific serotonin antagonist ICI 169,369 on the pituitary hormone response to insulin-induced hypoglycaemia in humans.
To examine the role of serotonin in pituitary hormone release by studying the effect of a specific 5HT2 receptor antagonist, ICI 169,369, on the ACTH, prolactin, growth hormone and AVP response to insulin-induced hypoglycaemia in healthy humans. A double-blind, within-subject trial using a crossover design to compare the effect of placebo with two doses of ICI 169,369 on pituitary hormone responses to insulin-induced hypoglycaemia. Ten healthy subjects were studied in the low-dose (30 mg x 2) limb and 11 healthy volunteers in the high-dose (80 mg x 2) limb. Plasma concentrations of prolactin, growth hormone, ACTH, cortisol and AVP, and blood glucose. In the low-dose study, pretreatment with 30 mg ICI 169,369, 10 and 2 hours before the study, had no effect on the fall in blood glucose or the rise in plasma ACTH, prolactin, growth hormone, AVP or plasma cortisol following insulin injection, when compared with placebo. In the high-dose study the effect of a higher dose (80 mg) of ICI 169,369 on the pituitary hormone response to hypoglycaemia was compared with that of placebo. Although the fall in blood glucose was similar following drug (4.3 +/- 0.1 to 1.5 +/- 0.5 mmol/l, mean +/- SEM, P less than 0.001) and placebo (4.3 +/- 0.1 to 1.4 +/- 0.4 mmol/l, P less than 0.001), the rise in plasma AVP was lower (P less than 0.05) following pretreatment with drug (0.5 +/- 0.2 to 2.1 +/- 0.6 pmol/l, P less than 0.05) than with placebo (0.7 +/- 0.2 to 3.4 +/- 0.9 pmol/l, P less than 0.01). The ACTH, prolactin, growth hormone and cortisol responses were unaffected by ICI 169,369. The data are compatible with an inhibitory effect of the serotonin antagonist ICI 169,369 on the AVP, but not the ACTH, prolactin or growth hormone response to insulin-induced hypoglycaemia in humans.


Euploid blastocyst rates in patients from POSEIDON groups 3 and 4 using propensity score matching.
Do patients with low ovarian reserve, as defined by the patient-oriented strategies encompassing individualized oocyte number (POSEIDON) criteria, have low euploid blastocyst rates? Retrospective study of 548 IVF cycles of patients with unexplained recurrent miscarriage who underwent preimplantation genetic test for aneuploidy (PGT-A). Euploid blastocyst rates were analysed to compare patients from POSEIDON groups 3 and 4 (serum anti-Müllerian hormone [AMH] levels <1.2 ng/ml) with those who have normal ovarian reserve (AMH levels ≥1.2 ng/ml) before and after using propensity score matching to match selected variables, such as female age, body mass index, the number of clinical miscarriages, ovarian stimulation protocols and PGT-A analysis platforms. Cycles of patients from POSEIDON groups 3 and 4 were then divided into four groups according to median and quartiles of serum AMH levels: <0.668 ng/ml, 0.668-0.890 ng/ml, >0.890-1.070 ng/ml and >1.070-<1.20 ng/ml. The euploid blastocyst rates were compared across these four groups. After using propensity score matching, no difference was found in euploid blastocyst rates between patients from POSEIDON groups 3 and 4 and those with normal ovarian reserve. Among cycles of patients from POSEIDON groups 3 and 4, no difference was found in euploid blastocyst rates between the different AMH levels. The decline in ovarian reserve in patients from POSEIDON groups 3 and 4 was not related to low euploid blastocyst rates. Serum AMH levels do not seem to be a predictor of euploid blastocyst rates in such patients.


Limits to sustained energy intake IX: a review of hypotheses.
Several lines of evidence indicate that animals in the wild may be limited in their maximal rates of energy intake by their intrinsic physiology rather than food availability. Understanding the limits to sustained energy intake is important because this defines an envelope within which animals must trade-off competing activities. In the first part of this review, we consider the initial ideas that propelled this area and experimental evidence connected with them. An early conceptual advance in this field was the idea that energy intake could be centrally limited by aspects of the digestive process, or peripherally limited at the sites of energy utilisation. A model system that has been widely employed to explore these ideas is lactation in small rodents. Initial studies in the late 1980s indicated that energy intake might be centrally limited, but work by Hammond and colleagues in the 1990s suggested that it was more likely that the limits were imposed by capacity of the mammary glands, and other works tended to support this view. This consensus, however, was undermined by studies that showed milk production was higher in mice at low temperatures, suggesting that the capacity of the mammary gland is not a limiting factor. In the second part of the review we consider some additional hypotheses that might explain these conflicting data. These include the heat dissipation limits hypothesis, the seasonal investment hypothesis and the saturated neural control hypothesis. Current evidence with respect to these hypotheses is also reviewed. The limited evidence presently available does not unambiguously support any one of them.


Repigmentation of hypopigmented scars using an erbium-doped 1,550-nm fractionated laser and topical bimatoprost.
Hypopigmented scarring is a challenging condition to treat, with current treatments showing limited efficacy and temporary results. Nonablative fractional resurfacing has been demonstrated to be an effective and safe modality in the treatment of hypopigmented scars. To demonstrate the efficacy and safety of combining fractional resurfacing with topical bimatoprost and topical tretinoin or pimecrolimus for the treatment of hypopigmented scars. Fourteen patients with hypopigmented scars were treated with a mean of 4.5 sessions of a fractionated 1,550-nm erbium-doped laser at 4- to 8-week intervals and subsequently started topical bimatoprost and tretinoin or pimecrolimus. An independent physician evaluated digital photographs taken before and 4 weeks after the last laser treatment using a quartile grading scale (grade 1, ≤25% improvement; grade 2, 26-50% improvement; grade 3, 51-75% improvement; grade 4, >75% improvement). Five patients had >75% improvement in hypopigmentation, and 12 had >50% improvement. After a mean follow-up of 20.1 months, all patients demonstrated prolonged results. Side effects were limited to transitory post-treatment edema and erythema. The combination of fractional resurfacing, topical bimatoprost, and tretinoin or pimecrolimus is a potential effective resource for the treatment of hypopigmented scars, with long-lasting results.


Keloid formation resulting in acquired syndactyly of an initially normal web space following syndactyly release of an adjacent web space.
We present a case of acquired syndactyly secondary to keloid formation in a previously normal web space following release of syndactyly in the adjacent web space.


Genome-wide investigation of the heat shock transcription factor (Hsf) gene family in Tartary buckwheat (Fagopyrum tataricum).
Heat shock transcription factor (Hsfs) is widely found in eukaryotes and prokaryotes. Hsfs can not only help organisms resist high temperature, but also participate in the regulation of plant growth and development (such as involved in the regulation of seed maturity and affects the root length of plants). The Hsf gene was first isolated from yeast and then gradually found in plants and sequenced, such as Arabidopsis thaliana, rice, maize. Tartary buckwheat is a rutin-rich crop, and its nutritional value and medicinal value are receiving more and more attention. However, there are few studies on the Hsf genes in Tartary buckwheat. With the whole genome sequence of Tartary buckwheat, we can effectively study the Hsf gene family in Tartary buckwheat. According to the study, 29 Hsf genes of Tartary buckwheat (FtHsf) were identified and renamed according to location of FtHsf genes on chromosome after removing a redundant gene. Therefore, only 29 FtHsf genes truly had the functional characteristics of the FtHsf family. The 29 FtHsf genes were located on 8 chromosomes of Tartary buckwheat, and we found gene duplication events in the FtHsf gene family, which may promote the expansion of the FtHsf gene family. Then, the motif compositions and the evolutionary relationship of FtHsf proteins and the gene structures, cis-acting elements in the promoter, synteny analysis of FtHsf genes were discussed in detail. What's more, we found that the transcription levels of FtHsf in different tissues and fruit development stages were significantly different by quantitative real-time PCR (qRT-PCR), implied that FtHsf may differ in function. In this study, only 29 Hsf genes were identified in Tartary buckwheat. Meanwhile, we also classified the FtHsf genes, and studied their structure, evolutionary relationship and the expression pattern. This series of studies has certain reference value for the study of the specific functional characteristics of Tartary buckwheat Hsf genes and to improve the yield and quality of Tartary buckwheat in the future.


The integration host factor (IHF) affects the expression of the phosphate-binding protein and of alkaline phosphatase in Escherichia coli.
The genes encoding alkaline phosphatase (phoA) and the inducible inorganic phosphate transport system Pst (pstS,C,A,B,U) belong to the PHO regulon. Mutants of Escherichia coli lacking the global regulatory protein integration host factor (IHF) show an increased level of alkaline phosphatase and a decreased level of Pst. IHF binds weakly but specifically to a DNA fragment containing the promoter region of the pst operon but does not bind to a fragment that includes the promoter region of phoA. It is proposed that IHF is a positive regulator of the pst operon and as such controls indirectly the expression of phoA.


The Rieske/cytochrome b complex of Heliobacteria.
Heliobacteria have a Rieske/cytochrome b complex composed of a Rieske protein, a cytochrome b(6,) a subunit IV and a di-heme cytochrome c. The overall structure of the complex seems close to the b(6)f complex from cyanobacteria and chloroplasts to the exception of the di-heme cytochrome. We show here by biochemical and biophysical studies that a heme c(i) is covalently attached to the Rieske/cytochrome b complex from Heliobacteria. We studied the EPR signature of this heme in two different species, Heliobacterium modesticaldum and Heliobacillus mobilis. In contrast to the case of b(6)f complex, a strong axial ligand to the heme is present, most probably a protonatable amino acid residue.


[Importance of machine blood count in the diagnosis of infectious mononucleosis by EBV].
EBV serology is recommended for serological diagnosis of mononucleosis. As results of an automated differential blood count is available more quickly, possible differences between an EBV primary infection and a bacterial tonsillitis were investigated. A retrospective evaluation of absolute and relative lymphocyte and monocyte counts of n = 140 patients > 16 years from 01/2008 to 01/2019 (mean age 21.4 years, 51 % ♀, 49 % ♂) with suspected EBV infection was performed. The groups of a serologically confirmed or excluded EBV infection were compared. An automated differential blood count was available in n = 42 patients with primary EBV infection. Average lymphocyte count was 5.5 ± 2.6 giga/l. Patients with acute bacterial tonsillitis (n = 36) had significantly lower values with 1.6 ± 1.3 giga/l, p < 0.05. Equal results were found in relative lymphocyte counts (47.4 ± 17.9 vs. 12.8 ± 9.1 %, p < 0.05). For monocyte counts, neither absolute (1.2 ± 0.8 vs. 1.2 ± 0.6 giga/l, p = 0.617) nor relative (8.8 ± 3.6 vs. 9.8 ± 5.2 %, p = 0.746) monocyte counts showed significant differences. Increased lymphocyte counts in an automated differential blood count can be a first indication of primary EBV infection. Perhaps up to 30 % morphologically altered lymphocytes are increasingly counted correctly with modern hematology analyzers and no longer counted as monocytes. These results could be used to make decisions about further diagnosis (abdominal ultrasonography, ECG) and antibiotic therapy before results of EBV serology are available.


Publisher Correction: Identification of new DNA-associated proteins from Waddlia chondrophila.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Spectrum of T-DNA integrations for insertional mutagenesis of Histoplasma capsulatum.
Agrobacterium-mediated transformation is being increasingly used for insertional mutagenesis of fungi. To better evaluate its effectiveness as a mutagen for the fungal pathogen Histoplasma capsulatum, we analyzed a collection of randomly selected T-DNA insertion mutants. Testing of different T-DNA element vectors engineered for transformation of fungi showed that pBHt2 provides the highest transformation efficiency and the lowest rate of vector backbone carryover. Sixty-eight individual T-DNA integrations were characterized by recovery of T-DNA ends and flanking genomic sequences. The right border (RB) end of the T-DNA is largely preserved whereas the left border (LB) end is frequently truncated. Analysis of T-DNA insertion sites confirms the lack of any integration hotspots in the Histoplasma genome. Relative to genes, T-DNA integrations show significant bias towards promoter regions at the expense of coding sequences. With consideration for potential promoter interruption and the demonstrated efficacy of intronic insertions, 61 % of mapped T-DNA insertions should impair gene expression or function. Mapping of T-DNA flanking sequences demonstrates 67 % of T-DNA integrations are integrations at a single chromosomal site and 31 % of T-DNA integrations are associated with large-scale chromosomal rearrangements. This characterization of T-DNA insertions in mutants selected without regard to phenotype supports application of Agrobacterium-mediated transformation as an insertional mutagen for genome-based screens and functional discovery of genes in Histoplasma.


Validity and reproducibility of self-reported working hours among Japanese male employees.
Working long hours is a potential health hazard. Although self-reporting of working hours in various time frames has been used in epidemiologic studies, its validity is unclear. The objective of this study was to examine the validity and reproducibility of self-reported working hours among Japanese male employees. The participants were 164 male employees of four large-scale companies in Japan. For validity, the Spearman correlation between self-reported working hours in the second survey and the working hours recorded by the company was calculated for the following four time frames: daily working hours, monthly overtime working hours in the last month, average overtime working hours in the last 3 months, and the frequency of long working months (≥45 h/month) within the last 12 months. For reproducibility, the intraclass correlation between the first (September 2013) and second surveys (December 2013) was calculated for each of the four time frames. The Spearman correlations between self-reported working hours and those based on company records were 0.74, 0.81, 0.85, and 0.89 for daily, monthly, 3-monthly, and yearly time periods, respectively. The intraclass correlations for self-reported working hours between the two questionnaire surveys were 0.63, 0.66, 0.73, and 0.87 for the respective time frames. The results of the present study among Japanese male employees suggest that the validity of self-reported working hours is high for all four time frames, whereas the reproducibility is moderate to high.


Closing the circle of care: implementation of a web-based communication tool to improve emergency department discharge communication with family physicians.
Postdischarge emergency department (ED) communication with family physicians is often suboptimal and negatively impacts patient care. We designed and piloted an online notification system that electronically alerts family physicians of patient ED visits and provides access to visitspecific laboratory and diagnostic information. Nine (of 10 invited) high-referring family physicians participated in this single ED pilot. A prepilot chart audit (30 patients from each family physician) determined the baseline rate of paper-based record transmission. A webbased communication portal was designed and piloted by the nine family physicians over 1 year. Participants provided usability feedback via focus groups and written surveys. Review of 270 patient charts in the prepilot phase revealed a 13% baseline rate of handwritten chart and a 44% rate of any information transfer between the ED and family physician offices following discharge. During the pilot, participant family physicians accrued 880 patient visits. Seven and two family physicians accessed online records for 74% and 12% of visits, respectively, an overall 60.7% of visits, corresponding to an overall absolute increase in receipt of patient ED visit information of 17%. The postpilot survey found that 100% of family physicians reported that they were ''often'' or ''always'' aware of patient ED visits, used the portal ''always'' or ''regularly'' to access patients' health records online, and felt that the web portal contributed to improved actual and perceived continuity of patient care. Introduction of a web-based ED visit communication tool improved ED-family physician communication. The impact of this system on improved continuity of care, timeliness of follow-up, and reduced duplication of investigations and referrals requires additional study.


A case of pseudogout of the temporomandibular joint with giant cell reparative granuloma of the temporal bone.
Pseudogout, also known as calcium pyrophosphate dihydrate deposition disease (CPPD) with giant cell reparative granuloma (GCRG) of the temporal bone is a rare disease, which is very easy to misdiagnose. When two diseases occur simultaneously, the pathological tissue of diseases is closely associated, which complicates clinical representation and causes enormous difficulty in diagnosis and treatment. We report a case of CPPD of the temporomandibular joint accompanied by surrounding GCRG of temporal bone in a 62-year-old male.


Evidence of a high molecular weight form of acrosin in boar acrosomal extract.
Acrosomal extracts of freshly ejaculated and immediately processed boar spermatozoa were investigated to detect which and how many acrosin molecular forms were present. Electrophoretic analyses of the acrosomal extract showed the presence of only one, slowly migrating, acrosin molecular form. Enzyme-linked-immuno-electro-transfer blot revealed the molecular weight of this form to be about 66 kdalton. Preliminary electrophoretic analyses under nondenaturating conditions of the acrosomal extract previously treated with thermolysin suggested that the approximately 66 kdalton form gives rise to two comigrating acrosin molecular forms.


Bifunctional hairy silica nanoparticles as high-performance additives for lubricant.
Bifunctional hairy silica nanoparticles (BHSNs), which are silica nanoparticles covered with alkyl and amino organic chains, were prepared as high-performance additives for lubricants. Compared with hairy silica nanoparticles covered by a single type of organic chain, binary hairy silica nanoparticles exhibit the advantages of both types of organic chains, which exhibit excellent compatibility with lubricants and adsorbability to metal surfaces. Nanoparticles with different ratios of amino and alkyl ligands were investigated. In comparison to an untreated lubricant, BHSNs reduce the friction coefficient and wear scar diameter by 40% and 60%, respectively. The wear mechanism of BHSNs was investigated, and the protective and filling effect of the nanoparticles improved because of collaboration of amino and alkyl ligands.


Very delayed coronary stent fracture: A case report.
A stent fracture is an emerging complication of the coronary stents. There are numerous risk factors for stent fractures; which include forceful exaggerated motion in the atrioventricular groove seen in right coronary artery, long stent, an ostial lesion at the point of maximum curvature in a tortuous vessel, stent over-expansion, stent overlapping with different size stents, complex lesion after stenting of a totally occluded vessel, Cypher stent and a highly mobile segment causing high mechanical stress. Furthermore, chronic stretch at specific vessel sites as bends may lead to late occurrence of fracture. Here we report a case of 40-year-old male who had two overlapping Cypher stents (3.0 × 13 mm and 2.75 × 18 mm) deployed at mid left anterior descending artery 2 years earlier presented with progressive chest pain.


Impact of multiple liver resections prior to salvage liver transplantation on survival in patients with recurrent HCC.
Salvage liver transplantation (SLT) is a controversial technique that has been reported to be acceptable for the management of patients with recurrent hepatocellular carcinoma (HCC) after primary hepatic resection (HR). However, whether the number of times liver resection is performed has an impact on survival after SLT has not yet been reported. Retrospective study. The level of care is primary and the study was carried out at only 1 centre. The study included 59 patients who underwent SLT for HCC from September 2001 to December 2012. 51 patients underwent HR only once before SLT, while the remaining 8 patients underwent HR more than once before SLT (HR=2 [7], HR=3, [1]). In this study, the 1-year, 3-year and 5-year overall and tumour-free survival outcomes between the 2 groups were compared. There were no significant differences between patients who underwent HR once and those who underwent HR more than once with respect to overall or tumour-free survival after receiving SLT. The 1-year, 3-year and 5-year overall survival rates for patients who underwent HR once were 72.9%, 35.3% and 35.5% vs 50%, 50% and 50%, respectively (p=0.986), while the 1-year, 3-year and 5-year tumour-free survival rates for those who underwent HR more than once were 66.3%, 55.3% and 44.4% vs 40%, 40% and 40%, respectively (p=0.790). There was no significant difference in the survival rate of patients who underwent HR once before SLT and those who underwent HR more than once. This suggests that SLT is a reasonable choice for patients who suffer from recurrent HCC after HR. This is a retrospective study and no registry or number is required.


Reduction of acid effects on trace element determination in food samples by CH4 mixed plasma-DRC-MS.
A robust method for trace element determination in food samples by addition of methane to the plasma of a dynamic reaction cell mass spectrometer (CH(4) mixed plasma-DRC-MS) was developed. Addition of 3 mL min(-1) methane to Ar-plasma eliminates the signal suppressions of various elements (As, Se, Hg, etc.) due to the high concentration of nitric acid (10%, v/v). The CH(4)-Ar mixed plasma may compensate for the plasma cooling effects due to the highly concentrated nitric acid. The interfering polyatomic ions (40)Ar(12)C(+), (40)Ar(35)Cl(+) and (40)Ar(40)Ar(+) on (52)Cr(+), (75)As(+) and (80)Se(+) determination were removed effectively using the DRC with CH(4) as the reaction gas. The limits of quantification (LOQ, 10σ) were 0.35 ng g(-1), 0.07 ng g(-1), 0.35 ng g(-1), 0.07 ng g(-1), 0.15 ng g(-1), and 0.07 ng g(-1) for As, Cd, Cr, Hg, Pb and Se, respectively. The proposed method was applied to the determination of these trace elements in four food standard reference materials (NIST1577b, GBW10018, NIST1570a and GBW10016), and the results were in good agreement with the certified values.


Fracture of the clavicle in the newborn. An ultrasound diagnosis.
Forty-one cases of clavicle fracture in newborn babies were examined by both radiographic and ultrasonic methods. No substantial difference has been found between these two modalities. It is suggested that ultrasound should be the procedure of choice in the diagnosis of clavicle fracture. This is especially true in those cases where impaired movement of the arm is the only clinical sign.


A Detailed Protocol for Subcellular RNA Sequencing (subRNA-seq).
In eukaryotic cells, RNAs at various maturation and processing levels are distributed across cellular compartments. The standard approach to determine transcript abundance and identity in vivo is RNA sequencing (RNA-seq). RNA-seq relies on RNA isolation from whole-cell lysates and thus mainly captures fully processed, stable, and more abundant cytoplasmic RNAs over nascent, unstable, and nuclear RNAs. Here, we provide a step-by-step protocol for subcellular RNA-seq (subRNA-seq). subRNA-seq allows the quantitative measurement of RNA polymerase II-generated RNAs from the chromatin, nucleoplasm, and cytoplasm of mammalian cells. This approach relies on cell fractionation prior to RNA isolation and sequencing library preparation. High-throughput sequencing of the subcellular RNAs can then be used to reveal the identity, abundance, and subcellular distribution of transcripts, thus providing insights into RNA processing and maturation. Deep sequencing of the chromatin-associated RNAs further offers the opportunity to study nascent RNAs. Subcellular RNA-seq libraries are obtained within 5 days. © 2017 by John Wiley & Sons, Inc.


Impact of Catheter Ablation on Arrhythmia Burden in Patients With Shock-Resistant Persistent Atrial Fibrillation.
Persistent shock-resistant atrial fibrillation (AF) is a challenging entity, with modest results from catheter ablation according to conventional survival analysis. The aim of this study was to determine the effect of catheter ablation on atrial tachyarrhythmia (ATA) burden in persistent AF patients undergoing first-time ablation with the use of an implantable cardiac monitor (ICM). Patients with drug-resistant ongoing persistent AF and at least 1 previous failed cardioversion were implanted with an ICM 2 months before the procedure. All patients underwent pulmonary vein isolation with or without additional substrate ablation depending on the presence of self-terminating AF on ICM and left atrium size. Median AF burden before and after ablation, off antiarrhythmic medication, was determined from ICM recordings after review by 2 independent investigators. Sixty patients were recruited (mean age 66 ± 9 years, 70% male). Mean left atrial diameter was 48 ± 6 mm and median CHA2DS2VASc score was 2. Ten patients (17%) unexpectedly demonstrated self-terminating AF before ablation. The median burden of ATA before ablation was 100% (95% CI: 19.6%-100%), decreasing to 0% (95% CI: 0%-95.8%) after ablation during the post-blanking follow-up period (median reduction 100%; 95% CI: 4%-100%; P < 0.001). Twenty-seven patients (45%) experienced recurrent ATA during 12-month follow-up. In these patients, median burden before ablation was 100% (95% CI: 26.9%-100%), decreasing to 11.4% (95% CI: 0.35%-99.7%) after ablation (P < 0.001). Quality of life improved significantly from baseline, driven by lack of recurrence. Patient-tailored catheter ablation results in a significant reduction in ATA burden (off antiarrhythmic medication) in shock-resistant persistent AF patients using ICMs implanted 2-months pre-procedure. These data suggest that conventional arrhythmia-free survival analysis does not capture the true impact of catheter ablation in this challenging cohort.


Speckle tracking-derived mitral annular velocities predict mortality in patients with acute coronary syndrome.
Myocardial ischemia can impair myocardial relaxation and result in increased left ventricular (LV) diastolic pressure. Noninvasive measurements of mitral annular velocities have been used to evaluate LV diastolic pressure. We sought to determine whether mitral annular velocities, derived from novel speckle tracking echocardiography (STE), could predict mortality in patients with acute coronary syndrome (ACS). A total of 246 patients with ACS were retrospectively studied. STE was analyzed offline with the sample volume placed on septal, lateral, inferior, and anterior mitral annulus. Peak early (E') and late (A') diastolic velocities of the mitral annulus were measured and averaged from the four regions. Peak early diastolic mitral inflow velocity (E) was obtained using pulsed-wave Doppler. Lower E' (P = 0.03), lower A' (P = 0.001), higher E'/A' ratio (P = 0.007), and higher E/E' ratio (P = 0.003) were independently associated with increased risk of death with adjustment for clinical and echocardiographic variables over the follow-up period of 21 months. The optimal cutoff value of E/E' ratio derived from the receiver operating characteristic analysis for predicting death was 30 (area under the curve = 0.65). E/E' ratio greater than 30 was predictive of death in univariate (HR, 2.40; CI, 1.42-4.06; P = 0.001) and multivariate (adjusted HR, 1.91; CI, 1.09-3.32; P = 0.02) models. The measurements of mitral annular velocities by STE are predictive of mortality in patients with ACS.


Transdermal Transmission Blocking Vaccine for Malaria using a Solid-in-Oil Dispersion.
Malaria is a mosquito-borne infectious disease that is widespread in developing countries. Malaria vaccines are important in efforts to eradicate malaria; however, vaccines are usually administered by injection, which requires medical personnel and has a risk of causing infection. Transdermal vaccines can be administered without damaging the skin and thus are ideal for the prevention of malaria. However, the stratum corneum forms a "brick and mortar" like structure in which stratum corneum cells are embedded in a hydrophobic matrix composed of lipids, which strongly inhibits the permeation of hydrophilic substances. In the present study, we designed a transdermal vaccine against vivax malaria using a solid-in-oil (S/O) dispersion. The S/O dispersion of a transmission blocking vaccine candidate, Pvs25 from Plasmodium vivax, showed higher skin penetration than that of the aqueous solution. Mice immunized with the S/O dispersion generated antibodies at similar titers as the mice immunized by injection, over the mid- to long-term. These results provide information for the development of transdermally administered malaria vaccines toward the eradication of malaria.


Malignant Otitis External: Our Experience and Literature Review.
BACKGROUND Malignant external otitis (MEO) is an invasive infection that can involve the external auditory canal and the skull base up to the contiguous soft tissues. Considering the changing face of MEO, we reviewed cases of MEO treated in our Ear Nose Throat (ENT) clinic - University Federico II of Naples between 2018 and 2019 to evaluate the current epidemiology of the condition and to assess the state of art on diagnosis, therapeutic and follow-up management in our patients. CASE REPORT We present the cases of three male patients with Type 2 diabetes mellitus who complained of long-lasting otorrhea and pain, with clinical suspicion of MEO. In all cases, ear swab was positive for Pseudomonas aeruginosa. All our patients received a 6-week course of intravenous ciprofloxacin, piperacillin, and tazobactam, with rapid clinical symptoms improvement and complete recovery at 1-year follow-up. CONCLUSIONS MEO is difficult to treat due to the lack of standardized care guidelines. Patients with MEO often present with severe otalgia, edema, otorrhea, and facial nerve paralysis. Clinicians must suspect MEO in elderly diabetic and immunocompromised patients with persistent otalgia after external otitis. Imaging (computed tomography and magnetic resonance imaging) can play synergistic roles in the management of MEO. To evaluate eradication of the disease, clinicians have to assess clinical symptoms and signs as well as radiological imaging and inflammatory markers.


Eating disorders symptomatology and inpatient treatment experience in eating-disordered subjects.
This study aimed to examine whether eating disorder (ED) subjects with and without a history of inpatient treatment differ in the severity of symptoms as measured by the Eating Disorders Inventory (EDI-1). In a sample of 222 women (66 women with anorexia nervosa, 126 women with bulimia nervosa, and 30 women with eating disorders not otherwise specified), the history of inpatient treatment was assessed and the EDI was administered. Fifty percent of the participants had received previous inpatient treatment. The group with past inpatient treatment had significantly higher scores on the EDI subscales Body Dissatisfaction, Ineffectiveness, and Interpersonal Distrust than the group without inpatient treatment experience. ED subjects with and without inpatient treatment showed different profiles on the EDI subscales. The fact that these two groups differ in specific ED symptoms may represent a bias in ED studies when samples with heterogeneous treatment contact history are recruited.


Birth-related PTSD symptoms and related factors following preterm childbirth in Turkey.
Objective: To examine factors associated with birth-related post-traumatic stress disorder (PTSD) among women who had preterm birth in their last pregnancy in Turkey.Methods: 304 women were asked to report sociodemographic factors, perinatal factors, birth-related factors, preterm birth/premature infant characteristics, and social support factors and PTSD symptoms. Data were collected using online surveys between November 2020 and February 2021. Hierarchical multiple linear regression was used. Results: The prevalence of birth-related PTSD symptoms following preterm birth was 71.1%. Older age, the woman being positively affected by her own mother's birth experience, not having traumatic experience in pregnancy and in the postnatal period, lower stress level after traumatic events experienced during birth, not feeling that their life/physical integrity was at risk during birth, having amniotomy, feeling psychologically well after childbirth, not being negatively affected by witnessing other parents' happy moments with their babies in friend/family groups, the absence of infant illness and mother's reporting higher positive interactions with healthcare team were associated with decreased likelihood of birth-related PTSD. Except for age and traumatic event in the postnatal period, all the variables explained 43% of the variance with a small effect size (f 2 = 0.04). Stress level after the traumatic events experienced during labor was the strongest predictor of birth-related PTSD symptoms (β = 0.33). Conclusion: Wellbeing of mother and baby, facilitating interventions at labor, and positive communication with the healthcare team was associated with lower birth-related PTSD symptoms. The study findings highlighted on birth-related PTSD symptoms in mothers of preterm infants in Turkey.


Polycyclic aromatic hydrocarbon biomarkers and serum markers of inflammation. A positive association that is more evident in men.
Polycyclic aromatic hydrocarbons (PAHs) are potent atmospheric pollutants, occurring from anthropogenic and natural sources. Several animal studies have reported a positive association of PAHs with inflammation. However, it is not clear if lower background exposure to PAHs is associated with inflammation in humans, independent of smoking, a major source of PAHs. We examined participants from the National Health and Nutrition Examination Survey 2001-2002, 2003-2004, and 2005-2006. Our exposures of interest were eight urinary monohydroxy polycyclic aromatic hydrocarbon biomarkers. Our outcomes were serum markers of inflammation; C-reactive protein (CRP) (≤10 mg/L) and total white blood cell (WBC) count (4000-12,000 cells/μL). Compared to participants with summed biomarkers of low-molecular weight (LMW) PAHs in the lowest quartile, the multivariable odds ratios (95% confidence interval) of high serum CRP (≥3 mg/L) and high total WBC count (defined as at or above the 95 percentile of total WBC distribution) among participants in the highest exposure quartile were 1.77 (1.13, 2.76) and 1.34 (1.12, 1.60) respectively. Urinary 1-hydroxypyrene, the biomarker of the higher molecular weight pyrene, was positively associated with total WBC count, and to lesser extent with serum CRP. In subsequent analyses, the positive association between LMW PAHs and serum CRP and total WBC count was found to be present within the stratified subgroups, independent of smoking and other potential confounders. The positive association was more evident among adult males when compared to females. Urinary PAH biomarkers were found to be positively associated with serum CRP and total WBC count independent of smoking and other potential confounders. The association was more evident in men.


An assessment of the in vivo clastogenicity of erythrosine.
In an investigation of the in vivo clastogenic potential of the food colouring erythrosine (ER), male B6C3F1 mice were treated by ip injection at doses of 50, 100 and 200 mg/kg, repeated 24 hr apart. Signs of toxicity were observed at the highest dose of ER administered. The three cytogenetic endpoints analysed were sister chromatid exchanges (SCEs) in peripheral blood lymphocytes (PBLs), micronuclei in bone marrow polychromatic erythrocytes (PCEs), and micronuclei in peripheral blood reticulocytes (PBRs). SCE frequencies in PBLs were 4.13, 4.58, 4.33 and 4.60 SCE/cell at 0, 50, 100 and 200 mg ER/kg, respectively. At the same doses, the frequencies of micronucleated PCEs were 3.5, 3.2, 2.0 and 2.5/1000 PCEs. Micronuclei in PBRs ranged from 1.2 to 3.6 and from 1.4 to 3.0/1000 PBRs in control and treated mice, respectively. These results indicate that ER is inactive as a clastogen in mouse blood and marrow cells. This result supports the hypothesis of a non-genotoxic mechanism for ER carcinogenicity.


Fluorescence sensor for organophosphorus pesticide detection based on the alkaline phosphatase-triggered reaction.
The threat of organophosphorus pesticide (OPP) residue to food safety and human health has caused widespread concern. In this paper, a sensitive fluorescence sensor for OPP detection was constructed based on the alkaline phosphatase (ALP) -triggered in situ reaction. In this method, ALP catalyses the dephosphorylation of the substrate l-ascorbic acid 2-phosphate sesquimagnesium salt hydrate (AAP) to generate l-ascorbic acid (AA). AA instantly combines with o-phenylenediamine (OPD) to form 3-(1,2-dihydroxyethyl)furo[3,4-b]quinoxalin-1(3H)-one (DFQ), which contains a quinoxaline core skeleton fluorophore and emits a strong fluorescence intensity at 425 nm. The existence of OPPs inhibits the activity of ALP and the production of AA and DFQ. As a result, the fluorescence intensity obviously decreases. Under optimal conditions, the fluorescence intensity linearly depends on the logarithm of chlorpyrifos concentration over a wide range of 20 pg/mL ∼1000 ng/mL with a detection limit of 15.03 pg/mL (S/N = 3), which is lower than the previously reported values. The sensor with its satisfactory accuracy and precision has been successfully applied to the detection of chlorpyrifos in leeks and celery samples with recoveries of 94.5-106.7% and an inter-assay relative standard deviation (RSD) below 11.51%. OPPs can be semiquantitatively determined by the colour changes in ultraviolet light. The superiority of the sensor is due to its visual simplicity without complex fluorescence labelling procedures and costly instruments.


Illness behaviour and general practice utilisation: a prospective study.
The Illness Behaviour Questionnaire (IBQ) was used to compare general practice patients who presented physical complaints in the absence of objective pathology, with those in whom the presence of pathology was established. Patients without pathology showed a greater conviction as to the presence of disease, and greater degrees of anxiety, depression and irritability. Males and females differed on their IBQ scores: males showing more disease conviction, somatic focusing and hypochondriasis. Utilisation of general practitioner services (as indicated by the number of visits in the six months subsequent to completing the IBQ) was associated with greater age, and for the group as a whole, utilisation was predicted by higher scores on the following IBQ scales: disease conviction, affective disturbance and disease affirmation. This was also the case for males, but in females only affective disturbance correlated with a greater number of visits. Four patterns were delineated in the relationship between age, illness behaviour variables, the presence or absence of objective pathology, and G.P. contacts.


Influence of the intracellular GluN2 C-terminal domain on NMDA receptor function.
Excitatory neurotransmission mediated by N-methyl-d-aspartate receptors (NMDARs) is fundamental to learning and memory and, when impaired, causes certain neurological disorders. NMDARs are heterotetrameric complexes composed of two GluN1 [NR1] and two GluN2(A-D) [NR2(A-D)] subunits. The GluN2 subunit is responsible for subunit-specific channel activity and gating kinetics including activation (rise time), peak open probability (peak Po) and deactivation (decay time). The peak Po of recombinant NMDARs was recently described to be controlled by the extracellular GluN2 N-terminal domain (NTD). The cytoplasmic GluN2 C-terminal domain (CTD) could also be involved, because the Po of synaptic NMDARs is reduced in mice expressing C-terminally truncated GluN2 subunits. Here, we examined the role of the GluN2 cytoplasmic tail for NMDAR channel activity and gating in HEK-293 cells. C-terminal truncation of GluN2A, GluN2B or GluN2C did not change the subunit-specific rise time but accelerated the decay time of glutamate-activated currents. Furthermore, the peak Po was reduced by about 50% for GluN2A and GluN2B but not for GluN2C. These results indicated that the CTD of GluN2 has a modulating role in NMDAR gating even in the absence of interacting synaptic proteins. Reduction of peak Po and deactivation kinetics following GluN2 C-terminal truncation were reversed by re-introducing a CTD from a different GluN2 subunit. Thus, the CTDs of GluN2 subunits behave as constitutive structural elements required for normal functioning of NMDARs but are not involved in determining the subunit-specific gating properties of NMDARs.


Determinants of defaulter to full vaccination among children aged 12-23 months in Siraro district, West Arsi zone, Oromia, Ethiopia: a case-control study.
Vaccination is one of the most cost-effective global public health interventions to reduce childhood morbidity and mortality. Defaulters to full vaccination can put children at greater risk of acquiring vaccine-preventable disease outbreaks. The reason for not receiving full vaccination is not well explored, and hence, there is limited evidence about defaulters of vaccination in Ethiopia. To identify determinants of defaulter to full vaccination among children aged 12-23 months in Siraro District, West Arsi Zone, Oromia Region, Ethiopia. A community-based unmatched case-control study was conducted among children aged 12-23 months from March 20 to April 30, 2022, with a total sample size of 444 (148 cases and 296 controls). Cases were children aged 12-23 months who had missed at least one routine vaccination dose, while controls were children who had received all of the recommended routine vaccinations. Consecutive sampling and simple random sampling techniques were used to select representative cases and controls respectively. Data were collected using a structured questionnaire, entered into Epi-data version 4.6, and exported to Statistical Package for Social Sciences version 26 for analysis. Logistic regression was used to identify determinants of the defaulter to full vaccination and the variables with p-value < 0.25 were recruited for multivariable analysis, and an adjusted odds ratio with a 95% confidence interval and a p-value of ≤ 0.05 was used to declare the statistical significance of the association. Of the assessed determinants of the defaulter to full vaccination; inadequate knowledge of mothers/caretakers (AOR = 4.32, 95% CI:2.78-6.70), educational status of a father unable to read and write (AOR = 3.66, 95% CI:1.29-10.39), time to reach health facility ≥ 30 minutes (AOR = 2.45, 95% CI:1.51-3.97), not told about the type of vaccine received (AOR = 2.37, 95% CI;1.27-4.45), no parents discussion on vaccination (AOR = 2.16, 95% CI:1.24-3.79), home delivery (AOR = 2.43, 95% CI:1.39-4.25) and not participated in pregnant mother conference (AOR = 2.47, 95% CI = 1.35-4.49) were the identified determinants of the defaulter to full vaccination. Mother's' knowledge, father's education, place of delivery, time to reach a health facility, health workers who told the type of vaccine received, participation in pregnant mother conference, and parents' discussion on vaccination were the determinants of the defaulter to full vaccination status. Thus, the district health office should work on defaulters of vaccination by strengthening immunization service delivery and improving maternal knowledge on vaccination through pregnant mother conference participation.


Serum PCSK9 levels, but not PCSK9 polymorphisms, are associated with CAD risk and lipid profiles in southern Chinese Han population.
Genetic and environment factors affect the occurrence and development of coronary artery disease (CAD). Proprotein convertase subtilisin/kexin type 9 (PCSK9), has been investigated extensively in the field of lipid metabolism and CAD. We performed this case-control study to investigate the relationship between serum PCSK9 levels and PCSK9 polymorphisms and lipid levels and CAD risk in a southern Chinese population. A hospital-based case-control study with 1, 096 subjects, including 626 CAD patients and 470 controls, were conducted. Genotyping of PCSK9 polymorphisms was performed using polymerase chain reaction-ligase detection reaction (PCR-LDR) method. The frequencies of the AA, AG and GG genotypes of PCSK9 E670G polymorphism were 90.58, 9.27, and 0.16% in the CAD patients, compared with 88.72, 10.85 and 0.43% in the controls, respectively. No R46L variant was detected in this population. There were no significant differences in genotype and allele frequencies of PCSK9E670G polymorphism between the CAD group and the controls. Serum lipid levels were not significantly different in carriers with the G allele and those with the AA genotype. The median (QR) of PCSK9 concentration was 1205.00 ng/l (577.28-1694.13 ng/l) in cases and 565.87 ng/l (357.17-967.50 ng/l) in controls, respectively. Compared with controls, CAD patients had significantly higher PCSK9 levels (z = 4.559, P < 0.001). After adjusting for age, gender, essential hypertension, diabetic mellitus, smoking and lipid profiles, PCSK9 levels remain significantly associated with increased CAD susceptibility (OR = 1.002, 95% CI = 1.001-1.002, P < 0.001). The correlation analyses showed that serum PCSK9 levels were positively associated with triglyceride (TG), Apo B and atherogenic index of plasma (AIP) levels in controls. No significant association between the PCSK9 E670G polymorphism and serum PCSK9 levels was observed in the CAD group and the controls. The present study shows that serum PCSK9 levels, but not PCSK9 polymorphisms, are associated with CAD risk in Southern Chinese Han population, and that serum PCSK9 levels are positively associated with AIP.


The Other Opioid Crisis: Just Another Drug Shortage?
The cause of drug shortages is a complex issue. This commentary highlights the shortage of intravenous opioid medications for cancer patients, in light of the opioid overdose epidemic.


Inter-seasonal variation in nitrogen uptake rates of the eutrophic Cochin estuary and adjacent coastal Arabian Sea.
The study assessed the Cochin estuary and adjacent coastal Arabian Sea for their seasonal variation in nitrate (NO3-) and ammonium (NH4+) uptake rates by total and nano + picoplankton using the 15N tracer technique. The results suggested that the NO3- and NH4+ uptake rates in the Cochin estuary are higher than those in the adjacent coastal Arabian Sea. NO3- and NH4+ uptake rates in the nearshore stations in the off Cochin station were high, indicating the influence of the eutrophic estuary. NO3- and NH4+ uptake rates conducted in off Mangalore transect were significantly lower than those of the off Cochin as it does not have an exchange with eutrophic systems. The nano + picoplankton's contribution to the total DIN uptake rates in the Cochin estuary was 77-98 %, indicating the relevance of nano + pico phytoplankton in the N cycling of the region.


Normal force distribution and posture of a hand pressing on a flat surface.
Hand strength data are needed to understand and predict hand postures and finger loads while placing the hand on an object or surface. This study aims to analyze the effect of hand posture and surface orientation on hand force while pressing a flat surface. Twelve participants, 6 females and 6 males ages 19-25, performed three exertions (100%, 30% and 10% MVC- Maximum Voluntary Contraction) perpendicular to a plate in 4 angles (-45°, 0°, 45° and 90° with respect to the horizontal plane) at elbow height. Exertions involved pushing in two postures: (1) whole hand and (2) constrained to only using the fingertips. Inter-digit joint angles were recorded to map hand and finger motions and estimate joint moments for each condition. Participants exerted twice the force when pushing with whole hand vs. fingertips. 72-75% of the total force was exerted over the base of the palm, while only 11-13% with the thumb for exertions at 90°, 45° or 0° plate angles. Males maximum force for pushing at 0°, 45° and 90° plates averaged 49% higher than females for the whole hand and 62% for the fingertips (p < 0.01). There was no significant sex difference (p > 0.05) for the -45° plate. Thumb joint loads were generally higher than the other individual fingers (p < 0.05) in all % MVC and accounted for 12% of total force during whole hand exertions. On average, joint moments were 30% higher during fingertip conditions vs. whole hand. Thumb and finger joint moment magnitudes when pushing the plate at 100% MVC indicated that Metacarpophalangeal (MCP) joint moments were higher (p < 0.05) than Distal Interphalangeal joints (DIP) and Proximal Interphalangeal joints (PIP) under whole hand and fingertips conditions.


Lower mitochondrial respiration does not lead to decreased fat oxidation in young African American women without obesity.
The prevalence of type 2 diabetes in African American women (AAW) is nearly twice that of White women. Lower insulin sensitivity and decreased mitochondrial function may be contributing factors. The purpose of this study was to compare fat oxidation in AAW and White women. Participants were 22 AAW and 22 White women, matched for age (18.7-38.3 years) and BMI (< 28 kg/m2). Participants completed two submaximal (50% VO2max) exercise tests with indirect calorimetry and stable isotope tracers to assess total, plasma, and intramyocellular triglyceride fat oxidation. The respiratory quotient during the exercise test was nearly identical in AAW and White women (0.813 ± 0.008 vs. 0.810 ± 0.008, p = 0.83). Although absolute total and plasma fat oxidation was lower in AAW, adjusting for the lower workload in AAW eliminated these racial differences. There was no racial difference in plasma and intramyocellular triglyceride source of fat for oxidation. No racial differences were observed in rates of ex vivo fat oxidation. Exercise efficiency was lower in AAW when adjusted to leg fat free mass. The data suggest that fat oxidation is not lower in AAW compared with White women, but additional studies are needed across exercise intensity, body weight, and age to confirm these results.


Hydrophilic and hydrophobic deep eutectic solvent-based extraction to determine parathion in cereals by digital image colorimetry integrated with smartphones.
To determine parathion in cereals, hydrophilic and hydrophobic deep eutectic solvents (DESs) were used by digital image colorimetry with smartphones. In the solid-liquid extraction part, hydrophilic DESs were used as extractants to extract parathion from cereals. In the liquid-liquid microextraction part, hydrophobic DESs dissociated into terpineol and tetrabutylammonium bromide in situ. The dissociated hydrophilic tetrabutylammonium ions reacted with parathion extracted in hydrophilic DESs under alkaline conditions to produce a yellow product, which was extracted and concentrated by dispersed organic phase terpinol. Digital image colorimetry integrated with the use of a smartphone was used for quantitative analysis. The limit of detection and quantification were 0.003 mg kg-1 and 0.01 mg kg-1, respectively. The recoveries for parathion were 94.8-106.2% with a relative standard deviation less than 3.6%. The proposed method was applied to analyze parathion in cereal samples: the method has the potential to be applied to pesticide residue analysis in food products.


Respiratory health effects associated with ambient sulfates and ozone in two rural Canadian communities.
A cross-sectional epidemiological study investigating the respiratory health of children in two Canadian communities was conducted in 1983-1984 in Tillsonburg, Ontario, located in a region of moderately elevated concentrations of transported air pollutants, and in Portage la Prairie, Manitoba, situated in a low pollution area. There were no significant local sources of industrial emissions in either community. Seven hundred and thirty-five children aged 7-12 were studied in the first town and 895 in the second. Respiratory health was assessed by the measurement of the forced vital capacity (FVC) and forced expiratory volume in 1 sec (FEV1.0) of each child, and by evaluation of the child's respiratory symptoms and illnesses using a parent-completed questionnaire. Sulfur dioxide (SO2), sulfate, and particulate nitrate levels were significantly higher in Tillsonburg than in Portage la Prairie (P less than 0.05), but nitrogen dioxide (NO2) and inhalable particles (PM10) differed little between the communities. Historical data in the vicinity of Tillsonburg indicated that average annual levels of sulfates, total nitrates, and ozone (O3) did not vary markedly in the 9-year period preceding the study. The results show that Tillsonburg children had statistically significant (P less than 0.001) lower levels of 2% for FVC and 1.7% for FEV1.0 as compared with children in Portage la Prairie. These differences could not be explained by parental smoking or education, the use of gas cooking or wood heating fuels, pollution levels on the day of testing, or differences in age, sex, height, or weight. The differences persisted when children with cough with phlegm, asthma, wheeze, inhalant allergies, or hospitalization before age 2 for a chest illness were excluded from analysis. With the exception of inhalant allergies, which occurred more frequently in Tillsonburg children, the prevalence of chronic respiratory symptoms and illnesses was similar in the two communities.


Effects of low molecular weight heparin on platelet surface P-selectin expression and serum interleukin-8 production in rats with trinitrobenzene sulphonic acid-induced colitis.
To observe the effects of low molecular weight heparin (LMWH) on platelet surface P-selectin expression and serum interleukin-8 production in rats with trinitrobenzene sulphonic acid (TNBS) induced colitis. Colitis was induced in female Sprague-Dawley rats by colonic administration of 2, 4, 6-TNBS. LMWH, a dalteparin (150 U/kg, 300 U/kg), was subcutaneously administrated one hour before induction of colitis and went on once a day for 6 days. Then a half dose was given for the next 7 days. Control animals received the same volume of normal saline once a day for 14 days after treated by TNBS. Animals were sacrificed at 24 h, days 7 and 14 after induction of colitis. The colon was excised for the evaluation of macroscopic and histological findings and TNF-alpha immunohistochemical assay. Platelet surface P-selectin expression was determined by radioimmunoassay and serum IL-8 production was assayed by ELISA method. LMWH treatment in a dose of 300 U/kg for 14 days significantly improved colonic inflammation by histological examination. Serum IL-8 production in the 300 U/kg treatment group was more significantly decreased at day 14 than that at 24 h (P<0.05). However, platelet surface P-selectin expression and TNF-alpha staining in colonic tissue were not significantly different among the three groups. LMWH has an anti-inflammatory effect on TNBS induced colitis in rats. The effect is possibly related to inhibition of proinflammatory cytokine IL-8, but not involved platelet surface P-selectin expression.


Structure of maltotetraose-forming amylase from Pseudomonas saccharophila STB07 provides insights into its product specificity.
The maltooligosaccharide-forming amylases (MFAses) degrade starch into maltooligosaccharides which potentially benefit human diet and grow popular in food processing, but little has been studied about their product specificity and structures. We focused on this topic and provide evidence through an X-ray crystal structure of the maltotetraose (G4)-forming amylase from Pseudomonas saccharophila STB07 (MFAps), as well as co-crystal structures of MFAps with G4 and with pseudo-maltoheptaose (pseudo-G7) determined at up to 1.1 Å resolution. G4 and pseudo-G7 occupy active cleft subsites -4 to -1 and -4 to +3 respectively. Binding induces conformational changes in the active sites except Asp193, working as the base catalyst. Comparison of the MFAps structure with those of other α-amylases revealed obvious differences in the loop structures providing dominant interactions between protein and substrate in the non-reducing side of the active sites cleft. These structures at the non-reducing end may govern the G4 specificity of MFAps and also be relevant to its exo-type action pattern.


A review of studies and measures to improve the mycotoxicological safety of traditional Japanese mold-fermented foods.
Miso (fermented soybean paste), shoyu (soy sauce) and sake (rice wine) are traditional moldfermented foods in Japan and have been consumed throughout much of its history. These have long been considered safe foods. In this contribution we review and summarize long-term studies to investigate potential problems with mycotoxin contamination of these products. The fungal cultures used for fermentation of these products are called "koji-molds" and mainly consist of strains ofAspergillus oryzae. A. oryzae belongs to theA. flavus group taxonomically, which is generally known to be a main producer of aflatoxins. Therefore, we studied the productivity of aflatoxins by various koji-molds, as well as the possibility of aflatoxin contamination of rice (which is used in the production of fermented foods), miso, shoyu and sake. Rice was found to be free from aflatoxins. Furthermore, none of the tested koji-molds produced any detectable levels of aflatoxins, consequently no aflatoxins were found in miso, shoyu, or sake. However, some koji-molds are known to produce cyclopiazonic acid (CPA) and kojic acid (KA). We studied the production of CPA and KA by various commercial koji-molds and identified some strains that produce relatively high amounts of CPA or KA. Consequently, we advised food industry not to use these strains. Although mycotoxin contamination of these products is therefore presently very low, further attempts should be made to completely eliminate CPA and KA from fermented foods.


Spot urine 5-hydroxyindoleacetic acid levels in the early diagnosis of acute appendicitis.
The significance of spot urine 5-hydroxyindoleacetic acid (5-HIAA) levels in the early diagnosis of acute appendicitis (AA) is questioned. Forty young, pigmented, male rabbits were divided into 4 groups. Groups are summarized as follows: group 1, control (n = 10); group 2, laparotomy, sham (n = 10); group 3, peritonitis with cecal ligation and puncture (n = 10); and group 4, experimental appendicitis (n = 10). Spot urine samples were obtained for 5-HIAA determination, and appendectomy materials were examined histopathologically and peritoneal fluid cultures were obtained after 12 hours in the study groups. The results were analyzed statistically with 1-way analysis of variance and Scheffé and Tukey's HSD tests. The mean 5-HIAA levels in the control group was 5.7 +/- 0.6 mg/L, whereas it is 5.9 +/- 0.9 mg/L, 6.6 +/- 0.6 mg/L, 9.4 +/- 0.9 mg/L in the sham, peritonitis, and appendicitis groups, respectively. The levels of 5-HIAA in the appendicitis group were higher than those in the other groups, which was statistically significant (P < .001). Spot urine 5-HIAA determination may be a practical, objective, and noninvasive method in the early diagnosis of AA. Because other radiological and laboratory tests can be insufficient, especially in the early period of AA, this test may be used effectively instead. Clinical studies are necessary for further conclusions.


Interdependency of stress relaxation and afferent nerve discharge in rat small intestine.
To be able to characterize intestinal mechano-electrical transduction, i.e. the mechanoreceptor behaviour, quantitative nerve studies with controlled and quantified stimulus are needed. This study aimed to determine the relationship between mechanical stress relaxation and afferent discharge adaptation evoked by fast isovolumetric bag distensions in the rat jejunum. Multiunit afferent activity was recorded in vivo from jejunum afferents from five male Wistar rats. The jejunum was distended via a bag at a distension speed of 0.5 ml/s to volumes of 0.2, 0.25, 0.3 and 0.4 ml, respectively. The distension was stopped and the volume was kept constant for 2 min to induce stress relaxation. The pressure in the bag, the afferent discharge (spike rate) and the diameter of the segment during the relaxation time were recorded simultaneously. The afferent discharge responses to distension showed a pattern with a peak during the sudden loading followed by decreasing activity with time. At distension volumes of 0.2, 0.25, 0.3 and 0.4 ml, the afferent discharge declined faster and to a greater extent (94%, 91%,96% and 87%) than the stress decreased (55%, 45%, 59% and 56%) during stress relaxation (p<0.001). Both the stress and the afferent discharge during the constant volume distension were independent of the distension volumes (p>0.5). The stress and the afferent discharge during the distension can be described mathematically on the basis of the quasi-linear theory of viscoelasticity. The association between the stress and the afferent discharge during the constant volume distension is linear with the same slope under various distension volumes. Intestinal mechanoreceptors were sensitive to the stress stimulus and a linear association between the stress relaxation and afferent discharge adaptation was found. The quasi-linear theory of visco-elasticity can be transferred to analysis of mechanical stimulus evoked afferent discharge.


[Measurement of retinal nerve fiber layer thickness of normal eyes with nerve fiber analyzer].
To study the retinal nerve fiber layer thickness (RNFLT) of normal eye and the correlation with age. RNFLT of 198 cases of normal eyes were examined by using nerve fiber analyzer (NFA). There were significant inverse linear correlation between the RNFLT and age (r = -0.46, P < 0.01). In the analysis by the quadrant, the RNFLT was correlated with age in the superior and inferior quadrant. The mean ratio of superior and inferior quadrant thickness (S/I) was 1.041. NFA seems to be an useful tool to measure the RNFLT quantitatively. The RNFLT of normal eye is found to be decreased with age.


Impact of the generation of EGFR-TKIs administered as prior therapy on the efficacy of osimertinib in patients with non-small cell lung cancer harboring EGFR T790M mutation.
There are few studies that directly compare the effects of osimertinib on patients with non-small cell lung cancer (NSCLC) harboring epidermal growth factor receptor (EGFR) T790M mutation between the generation of prior EGFR tyrosine kinase inhibitors (TKIs). We retrospectively reviewed clinical data from the medical records of consecutive patients with advanced NSCLC who had developed resistance to first- or second-generation EGFR-TKIs due to EGFR T790M mutation and were subsequently treated with osimertinib at Juntendo University Hospital. In patients with available tumor samples, target amplicon sequencing analyses were performed to explore the genetic biomarkers. A total of 38 patients with NSCLC harboring EGFR T790M mutation were treated with osimertinib. Eight patients were classified into group A (afatinib followed by osimertinib) and 30 patients were classified into group B (first-generation EGFR-TKI followed by osimertinib). Progression-free survival (PFS) was significantly longer in group A than in group B (median PFS; not reached vs. 11.0 months, P = 0.018). Fourteen patients had available tissue samples collected before osimertinib treatment for target sequencing. In group A we found no additional mutations, other than EGFR T790M mutation. On the other hand, there were three samples in which other mutations emerged, in addition to EGFR T790M mutation, in group B. PFS of osimertinib was significantly longer in patients with NSCLC harboring EGFR T790M mutation after treatment with afatinib than in patients after treatment with first generation EGFR-TKIs. Additional mutations other than EGFR T790M may affect the efficacy of osimertinib treatment. Significant findings of the study: PFS of osimertinib was significantly longer in patients with NSCLC harboring EGFR T790M mutation after treatment with afatinib than in patients after treatment with first generation EGFR-TKIs. Additional mutations other than EGFR T790M may affect the efficacy of osimertinib treatment.


Cold storage (8 C) of islets of Langerhans in a high-potassium "intracellular medium": near-normal insulin release and morphology after one week.
Islets of Langerhans from ob/ob mice were microdissected and stored for one week at 8 C or 37 C in a high-potassium "intracellular medium" or in a traditional culture medium (RPMI 1640 +/- fetal calf serum). Subsequent glucose-stimulated insulin release and islet morphology were best preserved by 8 C-storage in the high-potassium "intracellular medium". This suggests that normal, adult, mammalian tissues which do not readily divide in vitro may be better preserved for short periods of time at refrigerator temperatures in a high-potassium medium than by conventional culture at 37 C in a tissue culture medium.


Inadequate sleep moderates effects of interparental conflict appraisals on adolescent adjustment.
Previous research and theory suggest that interparental conflict that is perceived by youth as frequent/intense, threatening, and/or the responsibility of the youth is predictive of adjustment problems. In contrast, sleep, an important precursor to emotion regulation, is likely a protective factor for youth experiencing interparental conflict. The goal of the current study was to investigate the extent to which adolescents who maintain better and/or longer sleep are buffered from the adjustment problems associated with greater interparental conflict. One-hundred and fifty adolescents completed questionnaires assessing interparental conflict (frequency/intensity perceptions as well as threat and self-blame appraisals) as well as internalizing and externalizing behaviors. Participants also wore an actigraph over 1 week to objectively measure sleep quantity and quality; 123 participants had valid actigraphy data. Results of structural equation modeling analyses revealed that greater sleep quality weakened the association between interparental conflict and adjustment problems, particularly internalizing behaviors. However, sleep quantity was not a moderator. These results suggest that higher quality sleep may protect adolescents from the negative effects of interparental conflict.


Altered spatio-temporal state patterns for functional dynamics estimation in first-episode drug-naive major depression.
Patients with major depressive disorder (MDD) display affective and cognitive impairments. Although MDD-associated abnormalities of brain function and structure have been explored in depth, the relationships between MDD and spatio-temporal large-scale functional networks have not been evaluated in large-sample datasets. We employed data from International Big-Data Center for Depression Research (IBCDR), and comparable 543 healthy controls (HC) and 314 first-episode drug-naive (FEDN) MDD patients were included. We used a multivariate pattern classification method to learn informative spatio-temporal functional states. Brain states of each participant were extracted for functional dynamic estimation using an independent component analysis. Then, a multi-kernel pattern classification method was developed to identify discriminative spatio-temporal states associated with FEDN MDD. Finally, statistical analysis was applied to intrinsic and clinical brain characteristics. Compared with HC, FEDN MDD patients exhibited altered spatio-temporal functional states of the default mode network (DMN), the salience network, a hub network (centered on the dorsolateral prefrontal cortex), and a relatively complex coupling network (visual, DMN, motor-somatosensory and subcortical networks). Multi-kernel classification models to distinguish patients from HC obtained areas under the receiver operating characteristic curves up to 0.80. Classification scores correlated with Hamilton Depression Rating Scale scores and age at MDD onset. FEDN MDD patients had multiple abnormal spatio-temporal functional states. Classification scores derived from these states were related to symptom severity. The assessment of spatio-temporal states may represent a powerful clinical and research tool to distinguish between neuropsychiatric patients and controls.


Spectroscopic demonstration of a linkage between the kinetics of NTP hydrolysis and the conformational state of the recA-single-stranded DNA complex.
We recently constructed a mutant recA protein in which His-163 was replaced by a tryptophan residue; the [H163W]recA protein is functionally identical to the wild-type protein, and the Trp-163 side chain serves as a reporter group for the conformational transitions of the [H163W]recA-single-stranded DNA (ssDNA) complex. We have now examined the fluorescence properties of the [H163W]recA-ssDNA complex in the presence of a series of alternate nucleoside triphosphate cofactors. Under standard conditions (pH 7.5), ATP (S0.5 = 70 microM) and purine riboside triphosphate (PTP) (S0.5 = 110 microM) effect a 44% decrease in Trp-163 fluorescence and are active as cofactors for the DNA strand exchange reaction. In contrast, ITP (S0.5 = 400 microM) elicits only a 20% decrease in Trp-163 fluorescence (a level identical to that observed with the nucleoside diphosphates ADP, PDP, and IDP) and is inactive as a strand exchange cofactor. If the S0.5 (PTP) is increased to 130 microM (by increasing the pH of the reaction solution), the PTP-mediated quenching of Trp-163 fluorescence decreases to 20%, and PTP becomes inactive as a strand exchange cofactor. These results provide direct evidence for a linkage between the S0.5 value of a nucleoside triphosphate and the conformational state of the recA-ssDNA complex, with an S0.5 of 100-120 microM or lower required for stabilization of the strand exchange-active conformation.


Endothelin receptor agonists and antagonists exhibit different dissociation characteristics.
Endothelins (ETs) are vasoconstricting peptides that bind to membrane receptors to initiate their physiological effects. This report compares the dissociation characteristics of selected ET agonists and antagonists, and studies the effects of any difference in dissociation characteristics on the potency of antagonists. Competition studies using various ET receptor ligands against [125I]ET-1 or [125I]ET-3 binding demonstrated that porcine cerebellum membranes contain predominantly ETB receptor. [125I]IRL1620 associated with the receptors in a time-dependent manner. Although bound [125I]IRL1620 was easier to dissociate than bound [125I]ET-3, both agonists exhibited a dissociation half life > 20 h. For non-radiolabeled ligands, bind-and-wash studies were employed in which membranes were pre-incubated with unlabeled ligand followed by extensive washing before assaying for [125I]ET-1 binding. Results from bind-and-wash studies confirmed that bound non-radiolabeled IRL1620 and ET were as difficult to dissociate as [125I]ligands. In contrast, bound PD142893 and Ro46-2005 were easily dissociated from ETB receptors. Consequently, the inhibitory effects of PD142893 and Ro46-2005 on [125I]agonist binding diminished following incubation time. In cloned human ETA and ETB receptors, bound ET-1 was also more difficult to dissociate than bound antagonists. These results suggest that the differences in the dissociation characteristics of ET receptor agonists vs. antagonists may account for the diminished potency of Ro46-2005 and PD142893 as a function of incubation time.


Transrectal ultrasound and fine needle aspiration for malacoplakia of the prostate.
Clinically, malacoplakia of the prostate gland may mimic prostatic carcinoma. We report a case of prostatic malacoplakia in which transrectal ultrasound of the prostate was most compatible with carcinoma. However, fine needle aspiration cytology and biopsy revealed the classical histopathological features of malacoplakia so that a correct diagnosis could be made.


Effect of formulation parameters on 2-methoxyestradiol release from injectable cylindrical poly(DL-lactide-co-glycolide) implants.
The objective of this study was to investigate the potential of various formulation strategies to achieve 1-month continuous (improved) release of the novel anti-cancer drug, 2-methoxyestradiol (2-ME), from injectable cylindrical poly(DL-lactide-co-glycolide) (PLGA) implants. PLGA implants were prepared by a solvent extrusion method. PLGA 50:50 (M(w)=51 kDa, end group=lauryl ester) (PLGA-lauryl ester) implants loaded with 3-30 wt% 2-ME exhibited a pronounced lag phase (i.e., corresponding to induction time to polymer mass loss) and triphasic release profile. Incorporation of 5 wt% hydroxypropyl-beta-cyclodextrin (HP-beta-CD) (approximately 57% release after 28 days) or Pluronic F127 (approximately 42% release after 28 days) in PLGA-lauryl ester implants reduced the lag-phase and improved the drug release moderately over a period of 28 days. The formation and the incorporation of a 2-ME/polyethylene glycol (PEG) 8000 solid dispersion in PLGA-lauryl ester implants further increased drug release (approximately 21% and 73% release after 1 and 28 days, respectively), attributable to improved drug solubility/dissolution, higher matrix porosity, and accelerated polymer degradation. Blending of PLGA 50:50 (M(w)=24 kDa, end group=COOH) (PLGA-COOH) with the PLGA-lauryl ester also provided moderate enhancement of 2-ME release over a period of 28 days. PLGA-COOH (M(w)=24 kDa) implants with 3-5% w/w pore-forming MgCO(3) exhibited the most desirable drug release among all the formulations tested, and, demonstrated 1-month slow and continuous in vitro release of approximately 80% 2-ME after a minimal initial burst. Hence, these formulation approaches provide several possible avenues to improve release rates of the hydrophobic drug, 2-ME, from PLGA for future application in regional anti-cancer therapy.


Synthesis and antioxidant activity of novel amphipathic derivatives of tea polyphenol.
Hydrophobic derivatives of a tea polyphenol have been synthesized. 6, 8-Bis(octylthiomethyl)-epigallocatechin 3-O-gallate, 6, 8-bis(octylthiomethyl)-4 beta-(2-hydroxyethylthio)epigallocatechin 3-O-gallate and epigallocatechin 3-O-[4-O-(N-octadecylcarbamoyl)gallate] showed strong inhibition activity against lipid peroxidation of liposome caused by both lipid-soluble and water-soluble radical generators.


Signal recognition particle triggers the translocation of storage globulin polypeptides from field beans (Vicia faba L.) across mammalian endoplasmic reticulum membrane.
Hybridization-selected mRNAs coding for individual storage globulin polypeptides of field beans (Vicia faba L.) were translated in a cell-free system. Added mammalian signal recognition particle (SRP) recognizes cleavable signal peptides of the major vicilin and both legumin polypeptide precursors and induces translational arrest. The latter can be released by potassium-washed membranes (K-RM) leading to shortened polypeptides protected against proteases. Thus, SRP and K-RM function in a similar way with plant polypeptides as described for mammalian secretory proteins [(1981) J. Cell Biol. 91, 557-561]. Obviously, the initial steps in the biosynthesis and processing of plant storage globulin polypeptides are principally identical to those of animal secretory proteins.


Reference values for reticulocyte parameters and hypochromic RBC in healthy children.
New erythrocyte and reticulocyte parameters available on modern hematology analyzers have shown to be useful markers in children. However, pediatric method-specific reference ranges for these indices are sparse. We aim to establish pediatric reference values for reticulocyte parameters and hypochromic red blood cells in healthy children with the ADVIA 2120 hematology analyzer. Prospective study of 311 healthy children aged from 6 months to 18 years old with normal hematological parameters. ADVIA 2120 hematology analyzer (Siemens Healthcare Diagnostics, Amadora, Portugal) was used for whole-blood hematological measures. The sample population was grouped according to age into three cohorts, and gender distribution was used for participants aged ≥12 years. Age- and gender-specific reference limits of reticulocyte hemoglobin content (CHr or Ret-He), difference between the reticulocyte and erythrocyte hemoglobin content (Delta-He), immature reticulocyte fraction (IRF) and percentage of hypochromic red blood cells (%Hypo) are provided. Delta-He, IRF and %Hypo showed statistically significant gender differences in the 12-17 years group. CHr presented no significant gender variation within all age groups (median 30.8 ± 1.7 pg). Establishing reliable pediatric reference intervals for these novel hematological parameters may offer valuable prospects for clinical practice and research in the pediatrics field.


Denovo designing: a novel signal peptide for tat translocation pathway to transport activin A to the periplasmic space of E. coli.
The twin-arginine translocation (Tat) pathway is one of the bacterial secretory strategies which exports folded proteins across the cytoplasmic membrane. In the present study, we designed a novel Tat-signal peptide for secretion of human activin A used as a recombinant protein model here. In doing so, Haloferax volcanii, Halobacterium salinarum, and Escherichia coli Tat specific signal peptides were aligned by ClustalW program to determine conserved and more frequently used residues. After making the initial signal peptide sequence and doing some mutations, efficiency of this designed signal peptide was evaluated using a set of well-known software programs such as TatP, PRED-TAT, and Phobius. Then the best complex between TatC as an initiator protein in Tat secretory machine and the new designed signal peptide connected to activin A with the lowest binding energy was constructed by HADDOCK server, and ΔΔG value of - 5.5 kcal/mol was calculated by FoldX module. After that, efficiency of this novel signal peptide for secretion of human activin A to the periplasmic space of E. coli Rosetta-gami (DE3) strain was experimentally evaluated; to scrutinize the activity of the novel signal peptide, Iranian Bacillus Licheniformis α-Amylase enzyme signal peptide as a Sec pathway signal peptide was used as a positive control. The quantitative analysis of western blotting bands by ImageJ software confirmed the high secretion ability of the new designed signal peptide; translocation of 69% of the produced recombinant activin A to the periplasmic space of E. coli. Circular Dichroism (CD) spectroscopy technique also approved the proper secondary structure of activin A secreted to the periplasmic space. The biological activity of activin A was also confirmed by differentiation of K562 erythroleukemia cells to the red blood cell by measuring the amount of hemoglobin or Fe2+ ion using ICP method. In conclusion, this novel designed signal peptide can be used to secrete any other recombinant proteins to the periplasmic space of E. coli efficiently.


Food allergies and allergens: Characterization and perceptions among diagnosed food allergic individuals in Lebanon.
The aim of our study was to assess the knowledge, practices, and attitudes towards food allergens and allergies among diagnosed food allergic individuals in Lebanon. Seventy diagnosed participants were recruited after reaching out to all the allergists of the country. They completed in person or over the phone a comprehensive valid questionnaire composed of 49 questions. Wheat was reported as top food allergen (15.7%), while itchy skin and rash were the most reported symptoms (71.4% and 68.6%, respectively). Only 34 (48.6%) of participants indicated they carry medications, 58 (70.7%) were diagnosed using blood test, and 22 (31.4%) stated that they are very knowledgeable on the topic. In terms of knowledge, participants scored on average 84.2 ± 11.5%. In terms of best practices, participants scored on average 47.8 ± 28.3%. Having a health related educational background increased significantly (p < 0.05) both knowledge and best practices scores, while age and gender did not have an effect. Our results highlight the importance of organizing ongoing educational initiatives and emphasize the need to lobby policy makers to making allergen-warning labels obligatory in the country.


Elevated plasma levels of TL1A in newly diagnosed systemic lupus erythematosus patients.
Systemic lupus erythematosus (SLE) is an autoimmune disease. Cytokine-mediated immunity plays an important role in the pathogenesis of SLE. TNF-like ligand 1A (TL1A) belongs to the TNF superfamily of cytokines and has been found to perform significantly in autoimmune diseases, such as rheumatoid arthritis and inflammatory bowel disease. To date, no study has discussed the expression levels of TL1A in SLE. We found that plasma levels of TL1A were significantly higher in newly diagnosed SLE patients compared with controls. Correlation analysis showed that plasma levels of TL1A were positively associated with SLE disease activity index. These data indicated that TL1A may play a role in SLE and may reflect the disease activity for SLE.


Mapping a Syndemic of Psychosocial Risks During Pregnancy Using Network Analysis.
Psychosocial risks during pregnancy impact maternal health in resource-limited settings, and HIV-positive women often bear a heavy burden of these factors. This study sought to use network modeling to characterize co-occurring psychosocial risks to maternal and child health among at-risk pregnant women. Two hundred pregnant HIV-positive women attending antenatal care in South Africa were enrolled. Measured risk factors included younger age, low income, low education, unemployment, unintended pregnancy, distress about pregnancy, antenatal depression, internalized HIV stigma, violence exposure, and lack of social support. Network analysis between risk factors was conducted in R using mixed graphical modeling. Centrality statistics were examined for each risk node in the network. In the resulting network, unintended pregnancy was strongly tied to distress about pregnancy. Distress about pregnancy was most central in the network and was connected to antenatal depression and HIV stigma. Unintended pregnancy was also associated with lack of social support, which was itself linked to antenatal depression, HIV stigma, and low income. Finally, antenatal depression was connected to violence exposure. Our results characterize a network of psychosocial risks among pregnant HIV-positive women. Distress about pregnancy emerged as central to this network, suggesting that unintended pregnancy is particularly distressing in this population and may contribute to further risks to maternal health, such as depression. Prevention of unintended pregnancies and interventions for coping with unplanned pregnancies may be particularly useful where multiple risks intersect. Efforts addressing single risk factors should consider an integrated, multilevel approach to support women during pregnancy. ClinicalTrials.gov identifier: NCT03069417.


Isolation of an unusual strain of Edwardsiella tarda from turbot and establish a PCR detection technique with the gyrB gene.
The aim of this study was to report an unusual Edwardsiella tarda and develop an effective method to identify this bacterium. During the spring and summer of 2006, an epizootic occurred among cultured turbot (Scophthalmus maximus) in Qingdao, China. A gram-negative, rod-shaped bacterium (designated as LTB-4) was isolated from the infected fish, and was proved to be virulent to turbot. Based on the 16S rDNA sequencing and phenotypic tests, the bacterial pathogen was identified as E. tarda. Unlike those commonly described E. tarda strains, no flagellate was observed. Partial gyrB genes were amplified from E. tarda using the universal primers of gyrB genes and sequenced. The polymerase chain reaction (PCR) primers for the gyrB gene were designed specific to E. tarda. It revealed positive amplification of the gyrB fragment in E. tarda, whereas other bacterial species were negative. In addition, the technique enabled the recognition of E. tarda from diseased fish. The isolate was identified as E. tarda without flagellate and an effective method was developed to identify E. tarda based on using the gyrB gene as a taxonomic marker. The unusual E. tarda was first reported in China and the PCR allowed the rapid and sensitive detection of E. tarda.


Bicarbonate blocks iron translocation from cotyledons inducing iron stress responses in Citrus roots.
The effect of bicarbonate ion (HCO3(-)) on the mobilization of iron (Fe) reserves from cotyledons to roots during early growth of citrus seedlings and its influence on the components of the iron acquisition system were studied. Monoembryonic seeds of Citrus limon (L.) were germinated "in vitro" on two iron-deprived media, supplemented or not with 10mM HCO3(-) (-Fe+Bic and -Fe, respectively). After 21d of culture, Fe concentration in seedling organs was measured, as well as gene expression and enzymatic activities. Finally, the effect of Fe resupply on the above responses was tested in the presence and absence of HCO3(-) (+Fe+Bic or +Fe, respectively). -Fe+Bic seedlings exhibited lower Fe concentration in shoots and roots than -Fe ones but higher in cotyledons, associated to a significative inhibition of NRAMP3 expression. HCO3(-) upregulated Strategy I related genes (FRO1, FRO2, HA1 and IRT1) and FC-R and H(+)-ATPase activities in roots of Fe-starved seedlings. PEPC1 expression and PEPCase activity were also increased. When -Fe+Bic pre-treated seedlings were transferred to Fe-containing media for 15d, Fe content in shoots and roots increased, although to a lower extent in the +Fe+Bic medium. Consequently, the above-described root responses became markedly repressed, however, this effect was less pronounced in +Fe+Bic seedlings. In conclusion, it appears that HCO3(-) prevents Fe translocation from cotyledons to shoot and root, therefore reducing their Fe levels. This triggers Fe-stress responses in the root, enhancing the expression of genes related with Fe uptake and the corresponding enzymatic activities.


Risks of Aromatase Inhibitor-Related Cardiotoxicity in Patients with Breast Cancer in Asia.
Despite a preferred endocrine therapy for women with estrogen and progesterone receptor-positive breast cancer, aromatase inhibitors (AIs) have been reported to increase risks of cardiovascular events. Given that breast cancer patients in Asia are younger at diagnosis, it is urgent to investigate this safety concern. Through the Taiwanese National Cohort, we identified breast cancer patients initiating selective estrogen receptor modulators (SERMs) or AIs from 2010 to 2016. Outcomes includes major adverse cardio- and cerebrovascular events (MACCEs). The average follow-up duration was five years. We identified 16,730 breast cancer patients treated with SERMs and 11,728 receiving AIs. The population was older and had more comorbidities in the AI group than in the SERM group. After adjusting for age, cancer stage, cancer therapies, cardiovascular drugs and comorbidities, despite similar risks of MACCEs between AI and SERM users, the risk of HF was significantly higher in patients treated with SERMs after adjusted mortality as a competing risk. When divided by the age of 50 years, despite a similar MACCEs in the younger population, MACCEs remained significantly higher in the older population who received SERMs. In this Asian cohort, we found that among patients of old age or with advanced cancer stage, the use of SERMs was associated with a higher risk of cardiovascular events than the use of AIs.


Identification of a STAT4 binding site in the interleukin-12 receptor required for signaling.
The specificity of the various STAT SH2 domains for different tyrosine-containing peptides enables cytokines to activate different signaling pathways and to induce distinct patterns of gene expression. We show that STAT4 has a unique peptide specificity and binds to the peptide sequence pYLPSNID (where pY represents phosphotyrosine). This motif is found at tyrosine residue 800 in the beta2 subunit of the interleukin-12 receptor and is required for DNA binding and transcriptional activity of STAT4. Our data demonstrate that transfection of interleukin-12 receptor beta1 and beta2 subunits is sufficient for STAT4 activation but not for STAT1 or STAT3 activation.


[Proper Perioperative Handling of Continuous Medication in Patients with Cardiac Comorbidities].
Cardiac comorbidities place a significant burden on the German population. Every third adult is diagnosed with arterial hypertension (AHT). In 2017 congestive heart failure (CHF) pertained approximately 2,5 million of mandatory health-insured patients. Coronary artery disease (CAD) is diagnosed in 28,3% of men and 19,1% of women older than 65 years.For optimal perioperative care it is important to have a sound knowledge of current treatment strategies of cardiac comorbidities. This helps in gaining an optimal risk stratification of the individual patient. It also ensures an optimal anesthesiological perioperative care for the patient at hand. Recommendations for the perioperative discontinuation or continuation of cardiac active drugs vary between countries and responsible medical societies.This article provides an in-depth review of the current medical therapies for cardiac conditions like AHT, CHF or CAD. The varying recommendations for the perioperative discontinuation/continuation of these therapies are also reviewed.


Risk stratification for nonagenarians undergoing transcatheter aortic valve replacement.
There are disparate data on the outcomes of nonagenarians undergoing transcatheter aortic valve replacement (TAVR) compared with younger patients. The purpose of this study is to determine whether the Society of Thoracic Surgeons (STS) score can be used to identify the subset of nonagenarians that are at a significantly higher risk for poor postoperative outcomes after TAVR. A total of 425 patients above the age of 80 underwent elective TAVR between 12/2013 and 2/2018 and were included in this study. Patients were deemed intermediate or high risk based on an STS predicted the risk of surgical mortality score of 3% to 8% and more than 8%, respectively. Differences in postoperative outcomes and/or 6-month mortality between intermediate and high-risk octogenarians and nonagenarians were compared. Of the 425 patients, 112 (26.4%) patients were nonagenarians, and 313 (73.6%) patients were octogenarians. Fifty-four (48.2%) of the nonagenarians were stratified as high-risk, while 78 (24.9%) of the octogenarians were stratified as high-risk. There were no statistically significant differences in the composite outcomes between intermediate-risk nonagenarians and intermediate-risk octogenarians. In contrast, high-risk nonagenarians were significantly more likely to experience the composite outcome of major perioperative complications and/or 6-month mortality as compared to high-risk octogenarians. Intermediate-risk nonagenarians undergoing TAVR have similar postoperative outcomes compared to intermediate-risk octogenarians. However, high-risk nonagenarian patients undergoing TAVR experience significantly poorer outcomes compared to their octogenarian counterparts. Judicious patient selection for TAVR in this subgroup of patients is therefore warranted.


Projections of costs, financing, and additional resource requirements for low- and lower middle-income country immunization programs over the decade, 2011-2020.
The Decade of Vaccines Global Vaccine Action Plan has outlined a set of ambitious goals to broaden the impact and reach of immunization across the globe. A projections exercise has been undertaken to assess the costs, financing availability, and additional resource requirements to achieve these goals through the delivery of vaccines against 19 diseases across 94 low- and middle-income countries for the period 2011-2020. The exercise draws upon data from existing published and unpublished global forecasts, country immunization plans, and costing studies. A combination of an ingredients-based approach and use of approximations based on past spending has been used to generate vaccine and non-vaccine delivery costs for routine programs, as well as supplementary immunization activities (SIAs). Financing projections focused primarily on support from governments and the GAVI Alliance. Cost and financing projections are presented in constant 2010 US dollars (US$). Cumulative total costs for the decade are projected to be US$57.5 billion, with 85% for routine programs and the remaining 15% for SIAs. Delivery costs account for 54% of total cumulative costs, and vaccine costs make up the remainder. A conservative estimate of total financing for immunization programs is projected to be $34.3 billion over the decade, with country governments financing 65%. These projections imply a cumulative funding gap of $23.2 billion. About 57% of the total resources required to close the funding gap are needed just to maintain existing programs and scale up other currently available vaccines (i.e., before adding in the additional costs of vaccines still in development). Efforts to mobilize additional resources, manage program costs, and establish mutual accountability between countries and development partners will all be necessary to ensure the goals of the Decade of Vaccines are achieved. Establishing or building on existing mechanisms to more comprehensively track resources and commitments for immunization will help facilitate these efforts.


Successful twin pregnancy after embryo donation to a patient with XY gonadal dysgenesis.
A twin pregnancy was established in a patient with XY gonadal dysgenesis. The pregnancy was supported with exogenously administered hormones for the initial 100 days. The infants were delivered by emergency cesarean section at 35 weeks' gestation when severe preeclampsia developed in the mother.


Virtual Reality Assisted Motor Imagery for Early Post-Stroke Recovery: A Review.
Stroke is a serious neurological disease that may lead to long-term disabilities and even death for stroke patients worldwide. The acute period, ( ≤ 1 mo post-stroke), is crucial for rehabilitation but the current standard clinical practice may be ineffective for patients with severe motor impairment, since most rehabilitation programs involve physical movement. Imagined movement - the so-called motor imagery (MI) - has been shown to activate motor areas of the brain without physical movement. MI therefore offers an opportunity for early rehabilitation of stroke patients. MI, however, is not widely employed in clinical practice due to a lack of evidence-based research. Here, we review MI-based approaches to rehabilitation of stroke patients and immersive virtual reality (VR) technologies to potentially assist MI and thus, promote recovery of motor function.