Adverse cardiac effects of salt with fludrocortisone in hypertension.
The effect of salt on blood pressure (BP) is controversial. A more important question is whether salt can produce cardiac target-organ damage, irrespective of its effect on BP. We assessed the effect of salt with fludrocortisone on QT dispersion and echocardiographic left ventricular diastolic function in a prospective interventional study involving 29 hypertensive subjects with a raised aldosterone/renin ratio who were hospitalized for investigation of possible primary aldosteronism. Each subject over 4 days was given a total of 28.8 g (480 mmol) of sodium chloride and 1.5 mg of fludrocortisone with potassium supplementation. Baseline and posttreatment 12-lead ECGs and echocardiograms were obtained. There were no significant changes in body weight, pulse rate, or BP after treatment with salt and fludrocortisone. Plasma sodium was significantly increased from 141.4 (SD 2.1) to 142.6 (SD 2.4) mmol/L (P:=0.001). QT and QTc dispersion both significantly increased: +19.6 (SD 16.5) ms (95% CI, 13.4 to 25.9) (P:<0.001) and +19.8 (SD 20.9) ms (95% CI, 11.8 to 27.7) (P:<0.001), respectively. There were no significant changes in (n=15) left ventricular dimensions or systolic function, but all diastolic filling indexes, including the preload-independent index, flow propagation velocity (55.49 [SD 10.91] to 48.96 [SD 11.40] cm/s, P:=0.018) worsened, suggesting significant deterioration of left ventricular diastolic function with salt and fludrocortisone. In conclusion, a combination of salt with fludrocortisone increased QT dispersion and impaired left ventricular diastolic relaxation in hypertensive patients with high aldosterone/renin ratios. This raises the possibility that salt may have BP-independent adverse cardiac effects in susceptible hypertensive subjects.


Impact of Deficiency of Intrinsic Coagulation Factors XI and XII on Ex Vivo Thrombus Formation and Clot Lysis.
The contributions of coagulation factor XI (FXI) and FXII to human clot formation is not fully known. Patients with deficiency in FXI have a variable mild bleeding risk, whereas FXII deficiency is not associated with bleeding. These phenotypes make FXII and FXI attractive target proteins in anticoagulant therapy. Here, we studied the mechanisms of fibrin clot formation, stability, and fibrinolytic degradation in patients with severe FXI or FXII deficiency. Thrombin generation was triggered in platelet-poor (PPP) and platelet-rich plasma (PRP) with the biological FXII trigger sulfatides. Intrinsic and extrinsic thrombus formation and degradation in whole blood were determined with rotational thromboelastometry (ROTEM). Clot formation under flow was assessed by perfusion of whole blood over collagen microspots with(out) tissue factor (TF). Thrombin generation and clot formation were delayed in FXII- and FXI-deficient patients triggered with sulfatides. In FXI-deficient plasma, this delay was more pronounced in PRP compared to PPP. In whole blood of FXII-deficient patients, clots were smaller but resistance to fibrinolysis was normal. In whole blood of FXI-deficient patients, clot formation was normal but the time to complete fibrinolysis was prolonged. In flow chamber experiments triggered with collagen/TF, platelet coverage was reduced in severe compared with moderate FXI deficiency, and fibrin formation was impaired. We conclude that quantitative defects in FXII and FXI have a substantial impact on contact activation-triggered coagulation. Furthermore, FXI deficiency has a dose-dependent suppressing effect on flow-mediated and platelet/TF-dependent clot formation. These last data highlight the contribution of particularly FXI to hemostasis.


Patient Understanding of the Risks and Benefits of Biologic Therapies in Inflammatory Bowel Disease: Insights from a Large-scale Analysis of Social Media Platforms.
Few studies have examined inflammatory bowel disease (IBD) patients' knowledge and understanding of biologic therapies outside traditional surveys. Here, we used social media data to examine IBD patients' understanding of the risks and benefits associated with biologic therapies and how this affects decision-making. We collected posts from Twitter and e-forum discussions from >3000 social media sites posted between June 27, 2012 and June 27, 2015. Guided by natural language processing, we identified posts with specific IBD keywords that discussed the risks and/or benefits of biologics. We then manually coded the resulting posts and performed qualitative analysis using ATLAS.ti software. A hierarchical coding structure was developed based on the keyword list and relevant themes were identified through manual coding. We examined 1598 IBD-related posts, of which 452 (28.3%) centered on the risks and/or benefits of biologics. There were 5 main themes: negative experiences and concerns with biologics (n = 247; 54.6%), decision-making surrounding biologic use (n = 169; 37.4%), positive experiences with biologics (n = 168; 37.2%), information seeking from peers (n = 125; 27.7%), and cost (n = 38; 8.4%). Posts describing negative experiences primarily commented on side effects from biologics, concerns about potential side effects and increased cancer risk, and pregnancy safety concerns. Posts on decision-making focused on nonbiologic treatment options, hesitation to initiate biologics, and concerns about changing or discontinuing regimens. Social media reveals a wide range of themes governing patients' experience and choice with IBD biologics. The complexity of navigating their risk-benefit profiles suggests merit in creating online tailored decision tools to support IBD patients' decision-making with biologic therapies.


Synchrotron radiation circular dichroism of various G-quadruplex structures.
Here we report synchrotron radiation circular dichroism spectra of various G-quadruplexes from 179 to 350 nm, and a number of bands in the vacuum ultraviolet (VUV) are reported for the first time. For a tetramolecular parallel structure, the strongest band in the spectrum is a negative band in the VUV at 182 nm; for a bimolecular antiparallel structure with diagonal loops, a new strong positive band is found at 190 nm; for a bimolecular parallel structure with edgewise loops, a strong positive band at 189 nm is observed; and for a self-folded chair-type structure, the strongest band in the spectrum is a positive band at 187 nm. For the tetramolecular parallel structure, the CD signals at all wavelengths are dominated by contributions from quartets of G bases, and the signal strength is approximately proportional to the number of quartets. Our experiments on well-characterized G-quadruplex structures lead us to question past attributions of CD signals to helix handedness and G quartet polarity. Although differences can be observed in the VUV region for the various quadruplex types, there do not appear to be clear-cut spectral features that can be used to identify specific topological features. It is suggested that this is because a dominant positive band in the VUV seen near 190 nm in all quadruplex structures is due to intrastrand guanine-guanine base stacking. However, our spectra can serve as reference spectra for the G-quadruplex structures investigated and, not least, to benchmark theoretical calculations and empirical models.


Optimizing detection sensitivity on surface-enhanced Raman scattering of transition-metal electrodes with confocal Raman microscopy.
Some points on how to improve the detection sensitivity of confocal Raman microscopy for the study of surface-enhanced Raman scattering (SERS) of transition-metal electrodes are discussed, including the careful design of the spectroelectrochemical cell, proper selection of the thickness of the solution layer, the binning of charge-coupled device (CCD) pixels, and appropriate setting of the notch filter. Various roughening methods for the Pt, Rh, Fe, Co, and Ni electrode surfaces have been introduced in order to obtain SERS-active surfaces. It has been shown that the appropriate roughening procedure and the optimizing performance of the confocal Raman microscope are the two most important factors to directly generate and observe SERS on net transition-metal electrodes.


Alpha-(1-3)- and alpha-(1-6)-D-mannose-specific plant lectins are markedly inhibitory to human immunodeficiency virus and cytomegalovirus infections in vitro.
The alpha-(1-3)-D-mannose- and alpha-(1-6)-D-mannose-specific agglutinins (lectins) from Galanthus nivalis, Hippeastrum hybrid, Narcissus pseudonarcissus, and Listera ovata inhibited infection of MT-4 cells by human immunodeficiency virus types 1 and 2 (HIV-1 and HIV-2) and simian immunodeficiency virus at concentrations comparable to the concentrations at which dextran sulfate (molecular weight, 5,000 [DS-5000]) inhibits these viruses (50% effective concentration, 0.2 to 0.6 microgram/ml). Unlike DS-5000, however, the plant lectins did not inhibit the replication of other enveloped viruses, except for human cytomegalovirus (50% effective concentration, 0.9 to 1.6 microgram/ml). The plant lectins suppressed syncytium formation between persistently HIV-1- or HIV-2-infected HUT-78 cells and uninfected MOLT-4 (clone 8) cells at concentrations that were 5- to 10-fold lower than that required for DS-5000. Unlike DS-5000, however, the plant lectins did not inhibit HIV-1 binding to CD4+ cells. Combination of the plant lectins with DS-5000 led to a potent synergistic inhibition of HIV-1-induced cytopathogenicity in MT-4 cells and syncytium formation between HIV-infected HUT-78 cells and MOLT-4 cells. Our data suggest that alpha-(1-3)-D- and alpha-(1-6)-D-mannose-specific plant lectins interfere with an event in the HIV replicative cycle that is subsequent to the attachment of the virions to the cells (i.e., the fusion process).


Multicenter evaluation of AMPLICOR and automated COBAS AMPLICOR CT/NG tests for Neisseria gonorrhoeae.
The fully automated COBAS AMPLICOR CT/NG and semiautomated AMPLICOR CT/NG tests were evaluated in a multicenter trial for their ability to detect Neisseria gonorrhoeae infections. Test performance compared to that of culturing was evaluated for 2,192 matched endocervical swab and urine specimens obtained from women and for 1, 981 matched urethral swab and urine specimens obtained from men. Culture-negative, PCR-positive specimens that tested positive in a confirmatory PCR test for an alternative target sequence within the N. gonorrhoeae 16S rRNA gene were considered to be true positives. The overall prevalences of gonorrhea were 6.6% in women and 20.1% in men. The COBAS AMPLICOR and AMPLICOR formats yielded concordant results for 98.8% of the specimens and exhibited virtually identical sensitivities and specificities. The results that follow are for the COBAS AMPLICOR format. With the infected patient as the reference standard, the resolved sensitivities of PCR were 92.4% for endocervical swab specimens and 64.8% for female urine specimens. There were no significant differences in these rates between women with and without symptoms. Among symptomatic men, COBAS AMPLICOR sensitivities were 94.1% for urine and 98.1% for urethral swabs; for asymptomatic men, the results were 42.3 and 73.1%, respectively. In comparison, the sensitivities of culturing were 84.8% for endocervical specimens, 92.7% for symptomatic male urethral specimens, and only 46.2% for urethral specimens obtained from asymptomatic men. When PCR results were analyzed as if only a single test had been performed on a single specimen type, the resolved sensitivity was always higher. The resolved specificities of PCR were 99.5% for endocervical swab specimens, 99.8% for female urine specimens, 98.9% for male urethral swab specimens, and 99.9% for male urine specimens. The internal control revealed that 2.1% of specimens were inhibitory when initially tested. Nevertheless, valid results were obtained for 99.2% of specimens because 60.0% of the inhibitory specimens were not inhibitory when a second aliquot was tested. The COBAS AMPLICOR CT/NG test for N. gonorrhoeae exhibited high sensitivity and specificity with urethral swab and urine specimens from men and endocervical swab specimens from women and thus is well suited for diagnosing and screening for N. gonorrhoeae infection.


MCNTs@MnO2 Nanocomposite Cathode Integrated with Soluble O2-Carrier Co-salen in Electrolyte for High-Performance Li-Air Batteries.
Li-air batteries (LABs) are promising because of their high energy density. However, LABs are troubled by large electrochemical polarization during discharge and charge, side reactions from both carbon cathode surface/peroxide product and electrolyte/superoxide intermediate, as well as the requirement for pure O2. Here we report the solution using multiwall carbon nanotubes (MCNTs)@MnO2 nanocomposite cathode integrated with N,N'-bis(salicylidene)ethylenediaminocobalt(II) (CoII-salen) in electrolyte for LABs. The advantage of such a combination is that on one hand, the coating layer of δ-MnO2 with about 2-3 nm on MCNTs@MnO2 nanocomposite catalyzes Li2O2 decomposition during charge and suppresses side reactions between product Li2O2 and MCNT surface. On the other hand, CoII-salen works as a mobile O2-carrier and accelerates Li2O2 formation through the reaciton of (CoIII-salen)2-O22- + 2Li+ + 2e- → 2CoII-salen + Li2O2. This reaction route overcomes the pure O2 limitation and avoids the formation of aggressive superoxide intermediate (O2- or LiO2), which easily attacks organic electrolyte. By using this double-catalyst system of Co-salen/MCNTs@MnO2, the lifetime of LABs is prolonged to 300 cycles at 500 mA g-1 (0.15 mA cm-2) with fixed capacity of 1000 mAh g-1 (0.30 mAh cm-2) in dry air (21% O2). Furthermore, we up-scale the capacity to 500 mAh (5.2 mAh cm-2) in pouch-type batteries (∼4 g, 325 Wh kg-1). This study should pave a new way for the design and construction of practical LABs.


Periparturient endocrine changes of conceptus and maternal units in Holstein heifers bearing genetically different conceptuses.
Holstein heifers (n = 21) were balanced across sires and assigned to three service-sire-breed groups in which heifers were inseminated artificially to either purebred Angus (n = 7), Holstein (n = 7) or Brahman (n = 7) bulls. Semen from four bulls was used for each service sire-breed group. Blood samples were collected from a jugular vein thrice weekly from d 160 to 265 of pregnancy, daily thereafter until 15 d postpartum, and then thrice weekly until d 60 postpartum. Concentrations of progesterone, estrone, estradiol, and estrone sulfate from 23 d prepartum to parturition, and of 15-keto-13,14-dihydro-prostaglandin F2 alpha (PGFM) from 2 d prepartum to d 15 postpartum were measured by radioimmunoassay. Heifers within the Brahman-service-sire group had longer gestations (P less than .05) than those of Holstein- or Angus-service-sire groups (285.0 vs 278.7), 279.0 d). Calf birth weight was lower (P less than .05) in Angus- than Holstein- and Brahman-service-sire groups (30.6 vs 36.1, 43.4 kg). Daily trends of prepartum maternal progesterone concentrations were approximately 1 ng/ml lower (P less than .01) in Angus- than Holstein- or Brahman-service-sire groups until luteolysis occurred. Heifers bearing crossbred Angus conceptuses had lower daily trends of prepartum estrogens concentrations (P less than .01), whereas heifers of the Holstein- and, even more dramatically, of the Brahman-service-sire groups had a higher magnitude and greater rise of plasma estrogens concentrations between d -10 and -1 prepartum (less than .01). Postpartum mean concentrations (P less than .05) and response curves of PGFM were lower (P less than .01) in the Angus- than in the Holstein- or Brahman-service-sire groups. Calf birth weights were correlated with least-squares means for maternal concentrations of prepartum estrone (r = .57), estradiol (r = .59) and estrone sulfate (r = .64) and postpartum maternal concentrations of PGFM (r = .56). Functional responses of the conceptus (e.g., estrogens) and maternal units (e.g., progesterone and PGFM) were influenced by conceptus genotype during the periparturient period.


Revision cochlear implant surgery in adults and children is a safe and effective technique: a retrospective single faculty study in Okayama University Hospital, Japan.
Cochlear implant (CI) surgery is a safe surgical technique, although some patients require revision CI surgery. This study investigated the cause and underlying reason of revision CI surgery as well as hearing outcomes in a single institution. This retrospective study evaluated patients who underwent CI surgery between April 2006 to March 2022 (n = 351). Sex, aetiology of hearing loss (HL), age and period from initial CI surgery to reimplantation, cause of revision, and related factors were examined. Twelve patients (8 males, 4 females) received CI reimplantation. The revision surgery rate was 2.59% (3.15% children, 1.69% adults); the period from initial surgery to reoperation was 8.60 ± 6.56 years for 9 children with congenital HL and 15.27 ± 5.72 years for 3 adults with progressive HL. Device failure was the most common cause (n = 8), followed by infections (n = 2), advanced facial irritation symptoms (n = 1), and electrode slip-out (n = 1). Mean preoperative and postoperative CI thresholds were 44.0 ± 9.46 dBnHL and 39.19 ± 8.89 dBnHL (p < .068), respectively. Caregiver education, surgical technique advances, flap design, and extensive antibiotic use may decrease the revision surgery rate. The lack of post-revision deterioration of the hearing threshold contributed to well-being in patients with CI.


[Comparative characteristics of dyslipoproteinemias in different geographic zones of the USSR].
Total cholesterol, triglycerides, high and low density lipoprotein cholesterol were measured in 40- to 59-year-old males from Moscow, Tallinn, a mountain village in the Caucasus and settlements in the Chukotka. Mean values, and 10th and 90th percentiles were determined for each of the populations examined. The Moscow and Tallinn populations were found to be similar biochemically and differ from rural populations. Dyslipoproteinemias were more atherogenic in urban residents, as compared to their rural counterparts. It is suggested that dyslipoproteinemias have geographic and ethnic determinants that may contribute to the degree of atherogeny implicit in any type of lipid metabolic disorders.


Technological advancements for the detection of and protection against biological and chemical warfare agents.
There is a growing need for technological advancements to combat agents of chemical and biological warfare, particularly in the context of the deliberate use of a chemical and/or biological warfare agent by a terrorist organization. In this tutorial review, we describe methods that have been developed both for the specific detection of biological and chemical warfare agents in a field setting, as well as potential therapeutic approaches for treating exposure to these toxic species. In particular, nerve agents are described as a typical chemical warfare agent, and the two potent biothreat agents, anthrax and botulinum neurotoxin, are used as illustrative examples of potent weapons for which countermeasures are urgently needed.


A Randomized Comparison between Video Demonstration and Verbal Instruction in Improving Rota Haler Technique in Children with Persistent Asthma: A Pilot Study.
Inhalation therapy is the cornerstone in management of asthma. Failure to use the device properly is one of the factors incriminated in poor control of asthma. To compare the technique of rota haler use in children with persistent asthma immediately after receiving either verbal instruction or a video based demonstration and again at one month following intervention. A total of 28 children, older than six years attending the childhood asthma clinic of our hospital, who were prescribed rota halers for the first time and who were technique naïve were enrolled into the study, after obtaining informed consent from the parents after meeting inclusion and exclusion criteria. They were then randomly assigned to either verbal instructions (group of 14) or video demonstration group (group of 14), for teaching them the rota haler technique. Their technique was then assessed using a modified version of the inhaler device assessment tool immediately after education and one month after the intervention. The proportions of children with good technique in both groups at both time points were compared. There was no significant difference in the baseline variables like age, gender, location, socio-economic status and disease duration between both groups. The proportion of children achieving good technique was significantly more in the video group than the oral instruction group at immediate assessment and also at one month post intervention with an odds of 8 and 23.40 respectively (p=0.0262 and 0.0075 respectively). Video demonstration is effective in improving the technique of rota haler use at immediate assessment and at one month post intervention. Further studies are needed to validate this study and to assess factors that predict successful device use.


Impact of Primary Breast Surgery on Overall Survival of Patients With De Novo Metastatic Breast Cancer: A Systematic Review and Meta-Analysis.
Breast surgery in cases of de novo metastatic breast cancer (MBC) is associated with improved outcomes in retrospective studies, although the results of randomized controlled trials (RCTs) are conflicting. We aimed to investigate whether surgery in this context prolongs patient survival. We performed a systematic review of the literature to identify RCTs comparing surgery of primary breast cancer to no surgery in patients with de novo MBC. Cochrane Library, Embase, Medline (OVID), and Web of Science were searched with latest update in July 2023, while conference proceedings were manually searched. Data concerning patient and tumor characteristics, as well as outcomes, were extracted. A meta-analysis with random effects models was performed considering heterogeneity between trials. Overall, 3255 entries were identified and 5 RCTs fulfilled all inclusion criteria, which had enrolled 1381 patients. The overall estimation in the intention-to-treat population showed no benefit for patients who had surgical excision of the primary breast tumor (HR = 0.93; 95% CI, 0.76-1.14). No subgroups in terms of receptor status or patterns of metastasis seemed to benefit from surgery, except for younger/premenopausal patients (HR = 0.74, 95% CI, 0.58-0.94). Breast surgery was associated with improved local progression-free survival (HR = 0.37, 95% CI, 0.19-0.74). Surgery of the primary tumor in patients with de novo MBC does not prolong survival, except possibly in younger/premenopausal patients. Breast surgery should be offered within the context of well-designed clinical trials examining the issue.


The fluid silicone prosthesis.
Injected fluid silicone serves as a soft prosthetic device and is an effective alternative to surgery in the management of corns and calluses. The material can relieve pain in weightbearing scars, reduce the incidence of recurring neuropathic ulcers, and protect skin at points of bony pressure. Augmenting subcutaneous tissue reduces both the vertical forces that cause direct pressure and the shear stresses imposed on plantar skin during horizontal acceleration or deceleration. The only significant side effect has been fluid migration, which is asymptomatic upon weightbearing and is seen only rarely when small amounts are implanted. Histologic examination of long-term biopsy and necropsy specimens has revealed no adverse tissue response. The safety and efficacy of injectable silicone as a soft-tissue implant in the foot have not been confirmed by the Food and Drug Administration, and pending completion of authorized investigation, the material is not available. Successfully completed studies, federal approval, and proper use could provide relief for millions of people who suffer from these common foot disorders.


Equivalent pore estimate for the alveolar-airway barrier in isolated dog lung.
In high-pressure pulmonary edema, lung interstitial and air space edema liquids have equal protein concentrations (Am. J. Physiol. 231: 1466, 1976). This suggests that the alveolar-airway barrier separating the air and interstitial spaces is relatively unrestrictive, even without apparent epithelial injury. To estimate the equivalent pore population of the alveolar-airway barrier we inflated each of 18 isolated dog lung lobes for 1 h with a solution of colored tracer of uniform radius. Tracer radii ranged from 1.3 to 405 nm. After freezing the lobes in liquid N2, we measured interstitial tracer concentrations in frozen perivascular cuffs or in samples thawed after dissection from frozen cuffs. Relative to the concentrations instilled, interstitial concentrations ranged from 0.34 for the smallest particles (1.3 and 3.5 nm radius) to zero for particles with radii of 405 nm. From the results we designed a pore model of the alveolar-airway barrier to reproduce the concentrations we measured. No single-pore model could be obtained, although a three-pore model fit the data well. The model results predict that pores with radii of 1, 40, and 400 nm would account for 68, 30, and 2% of total liquid flux, respectively. The majority of liquid flux (68%) would occur through passageways smaller than the smallest tracer we used (1.3 nm radius). We believe the alveolar-airway barrier consists not only of tight intercellular junctions that allow passage of only water and electrolytes but also of a smaller number of large leaks that allow passage of particles up to nearly 400 nm in radius.


'The goddess that we serve': projecting international community at the first serial chemistry conferences, 1893-1914.
The emergence of conferences in the late nineteenth century significantly changed the ways in which the international scientific community functioned and experienced itself. In the early modern Republic of Letters, savants mainly related through print and correspondence, and apart from at local and later national levels, scholars rarely met. International conferences, by contrast, brought scientists together regularly, in the flesh and in great numbers. Their previously imagined community now became tangible. This paper examines how conferencing reshaped the collective of international scientists by zooming in on the massive meetings of the International Congress of Applied Chemistry, 1893-1914. Drawing on Emile Durkheim's studies of religious gatherings it analyses the ritualization of routine conference practices, such as plenary ceremonies, toasts, ladies' programmes and committee meetings. It looks at how roles were distributed as participants performed as hosts and guests, and in masculine and feminine and national and international identities. Importantly, it shows both how the sacralization of chemistry as a higher aim served to instil senses of dedication in order to organize labour and mitigate conflict, and how the self-perception of the international chemical community was based on contemporary understandings of parliament, democracy and representation.


Did the expansion of free GP care impact demand for Emergency Department attendances? A difference-in-differences analysis.
The removal of co-payments for General Practitioner (GP) services has been shown to increase utilisation of GP care. The introduction of free GP care may also have spillover effects on utilisation of other healthcare such as Emergency Department (ED) services, which often serve as substitutes for primary care, and where co-payments to attend exist for many. In Ireland, out-of-pocket payments are paid by the majority of the population to access GP care, and these costs are amongst the highest in Europe. However, in July 2015 all children in Ireland aged under 6 became eligible for free GP care. Using a large administrative dataset on 413,562 ED attendances between January 2015 and June 2016 we apply a difference-in-differences method, with treatment and control groups differentiated by age, to examine whether ED utilisation changed amongst younger children following the introduction of universal free GP care. In particular, we examine ED attendances following a GP referral, as referrals from GPs also afford access to the ED free of charge. We find that the expansion of free GP care did not reduce overall ED utilisation for under 6s. Additionally, we find that the proportion of ED attendances occurring through GP referrals increased by over 2 percentage points. This latter finding may be indicative of increased pressure placed on GPs from increased demand. Overall, this study finds that expanding free GP care to all young children did not reduce their ED utilisation.


Molecular genetic analysis of the hepatocyte growth factor/MET signaling pathway in pediatric medulloblastoma.
The hepatocyte growth factor (HGF)/MET pathway plays a critical role in the development of the nervous system and has been implicated in medulloblastoma pathogenesis. Recent studies have shown a role for gene amplification of activators of this pathway, as well as silencing of its inhibitors in medulloblastoma pathogenesis. We analyzed exon array data from a cohort of 103 primary medulloblastomas to show that HGF/MET pathway elements are dysregulated in tumors compared to normal cerebellum. To determine if mutation of HGF/MET pathway genes is a mechanism for pathway dysregulation, we conducted a mutational analysis by exon resequencing of three key components of this pathway, including serine protease inhibitor Kunitz-type 1 (SPINT1), serine protease inhibitor Kunitz-type 2 (SPINT2), and MET, in 32 primary human medulloblastoma specimens. From this analysis, we identified multiple coding synonymous and nonsynonymous single nucleotide polymorphisms in these genes among the 32 tumor samples. Interestingly, we also discovered two unreported sequence variants in SPINT1 and SPINT2 in two tumors that resulted in Arginine to Histidine amino acid substitutions at codons 418 and 233, respectively. However, conservation assessment and functional assays of these two variants indicate that they involve nonconserved residues and that they do not affect the function of SPINT1 and SPINT2 as tumor suppressor genes. In conclusion, our data suggest that mutation alone plays a minor role in causing aberrancies of the HGF/MET pathway in medulloblastoma in comparison with other malignancies such as breast, hepatocellular, renal, and lung carcinomas.


Clinical Application of the Internal Mammary Artery Perforator Adipofascial Flap.
Skin ulcers on the anterior chest wall are caused mainly by radiation therapy for breast cancer and anterior mediastinitis after thoracotomy, and they are often refractory. Some muscle flaps are commonly used for anterior chest wall reconstruction, but muscle flaps accompany high invasion. We used the internal mammary artery perforator (IMAP) adipofascial flap and IMAP skin flap for the anterior chest wall reconstruction. We examined the IMAPs using a handheld Doppler device and contrast-enhanced computerized tomography preoperatively. Each flap was designed based on the location of the IMAP and the size of the flap was dependent on the coverage required by the size and location of the skin ulcer. The location of the IMAPs functioned as the pivot point of the flap and the flap was flipped or swung on the defect. We used IMAP adipofascial flap for 2 cases and IMAP skin flap for 1 case. In those 3 cases, we could elevate the flap with no complications even after the internal mammary artery had been harvested. There was no recurrence of the skin ulcer or wound infection after the operation. In this study, we reported 3 cases of skin ulcer on the anterior chest wall reconstructed with the IMAP adipofascial and skin flap. To our knowledge, this is the first report of the use of the IMAP flap as an adipofascial flap. The IMAP adipofascial flap accompanies less invasion than muscle flaps and the surgical procedure is relatively easy. The IMAP adopofascial flap is considered as one of the effective means for anterior chest wall reconstruction.


Travel to the Persian Gulf. Medical guidelines for visitors.
The disease, destruction, and destitution created by the recent conflicts in the Persian Gulf have resulted in increased international travel to affected countries for relief and reconstruction. Such travel involves more than minimal health risks, but effective preparation of travelers by primary care physicians can help, as discussed by the authors.


Renal urea transporters. Direct and indirect regulation by vasopressin.
Urea is the most abundant urinary solute and is excreted in urine at a much higher concentration than in other body fluids. Urea concentration is achieved in the kidney through complex urea movements between blood vessels and renal tubules, which involve facilitated urea transport. Three major urea transporters expressed in the kidney have been cloned, UT-A1, UT-A2 and UT-B1, the first two derived from the same gene by differential transcription. These membrane proteins enable facilitated diffusion of urea through specific parts of the nephron (UT-A) and through renal vasculature (UT-B) in the medulla. UT-A1 is localised in the terminal part of the inner medullary collecting ducts and accounts for the vasopressin-dependent increase in urea permeability of this segment. UT-A2 is found in the descending thin limbs of Henle's loops. UT-B1 is expressed in the endothelium of the descending vasa recta supplying blood to the renal medulla, and in red cells. All three urea transporters are primarily involved in the process of intrarenal urea recycling, which enables the establishment, and prevents the dissipation, of a high concentration of urea in the inner medulla. This is an essential feature for producing a concentrated urine and thus for water economy in mammals. Vasopressin, upon binding to V2 receptors in the inner medullary collecting ducts, increases urea permeability through activation of UT-A1 molecules, thus enabling urea to diffuse into the inner medullary interstitium. Urea then taken up in ascending vasa recta is returned to the inner medulla via UT-A2 and UT-B1 by countercurrent exchange. These latter two urea transporters are not influenced acutely by vasopressin, but UT-A2 expression is markedly increased in the descending thin limbs of the loops of Henle after sustained exposure to vasopressin or its V2 agonist dDAVP. This effect is indirect because vasopressin receptors are lacking in the descending limbs. The acute direct and delayed indirect actions of vasopressin on renal urea transporters will increase medullary urea accumulation and thus the ability of the kidney to conserve water. Atrial natriuretic peptide inhibits the vasopressin-dependent increase in urea permeability in the inner medullary collecting ducts. The interruption of urea recycling probably contributes to the natriuresis. Impairing in this way the capacity of the kidney to concentrate urea enhances its capacity to concentrate sodium in the urine.


Clinical profiles of early and tuberculosis-related mortality in South Korea between 2015 and 2017: a cross-sectional study.
Although the incidence of tuberculosis (TB) has decreased in South Korea, the mortality rate remains high. TB mortality is a key indicator for TB control interventions. The purpose of this study was to assess early and TB-related mortality during anti-TB treatment and describe the associated clinical characteristics. A multicenter cross-sectional study was performed across South Korea. Patients with pulmonary TB who died during anti-TB treatment and whose records were submitted to the national TB surveillance system between 2015 and 2017 were enrolled. All TB deaths were categorized based on cause (TB-related or non-TB-related) and timing (early or late). We identified statistical associations using the frequency table, chi-square test, and binary logistic regression. Of 5595 notifiable mortality cases, 3735 patients with pulmonary TB were included in the analysis. There were 2541 (68.0%) male patients, and 2935 (78.6%) mortality cases were observed in patients older than 65 years. There were 944 (25.3%) cases of TB-related death and 2545 (68.1%) cases of early death. Of all cases, 187 (5.0%) patients were diagnosed post-mortem and 38 (1.0%) patients died on the first day of treatment. Low body mass index (adjusted odds ratio (aOR) = 1.26; 95% confidence interval (CI) = 1.08-1.48), no reported illness (aOR = 1.36; 95% CI = 1.10-1.68), bilateral disease on chest X-ray (aOR = 1.30; 95% CI = 1.11-1.52), and positive acid-fast bacilli smear result (aOR = 1.30; 95% CI = 1.11-1.52) were significantly associated with early death, as well as TB-related death. Acute respiratory failure was the most common mode of non-TB-related death. Malignancy was associated with both late (aOR = 0.71; 95% CI = 0.59-0.89) and non-TB-related (aOR = 0.35; 95% CI = 0.26-0.46) death. A high proportion of TB death was observed in elderly patients and attributed to non-TB-related causes. Many TB-related deaths occurred during the intensive phase, particularly within the first month. Further studies identifying risk factors for different causes of TB death at different phases of anti-TB treatment are warranted for early targeted intervention in order to reduce TB mortality.


Shear-Wave Elastography Assessments of Quadriceps Stiffness Changes prior to, during and after Prolonged Exercise: A Longitudinal Study during an Extreme Mountain Ultra-Marathon.
In sports medicine, there is increasing interest in quantifying the elastic properties of skeletal muscle, especially during extreme muscular stimulation, to improve our understanding of the impact of alterations in skeletal muscle stiffness on resulting pain or injuries, as well as the mechanisms underlying the relationships between these parameters. Our main objective was to determine whether real-time shear-wave elastography (SWE) can monitor changes in quadriceps muscle elasticity during an extreme mountain ultra-marathon, a powerful mechanical stress model. Our study involved 50 volunteers participating in an extreme mountain marathon (distance: 330 km, elevation: +24,000 m). Quantitative SWE velocity and shear modulus measurements were performed in most superficial quadriceps muscle heads at the following 4 time points: before the race, halfway through the race, upon finishing the race and after recovery (+48 h). Blood biomarker levels were also measured. A significant decrease in the quadriceps shear modulus was observed upon finishing the race (3.31±0.61 kPa) (p<0.001) compared to baseline (3.56±0.63 kPa), followed by a partial recovery +48 h after the race (3.45±0.6 kPa) (p = 0.002) across all muscle heads, as well as for each of the following three muscle heads: the rectus femoris (p = 0.003), the vastus medialis (p = 0.033) and the vastus lateralis (p = 0.001). Our study is the first to assess changes in muscle stiffness during prolonged extreme physical endurance exercises based on shear modulus measurements using non-invasive SWE. We concluded that decreases in stiffness, which may have resulted from quadriceps overuse in the setting of supra-physiological stress caused by the extreme distance and unique elevation of the race, may have been responsible for the development of inflammation and muscle swelling. SWE may hence represent a promising tool for monitoring physiologic or pathological variations in muscle stiffness and may be useful for diagnosing and monitoring muscle changes.


[Regulation of heliomycin biosynthesis by the carbon source].
The effect of different carbon sources on the biosynthesis of heliomycin by Streptomyces olivocinereus 11-98 was studied. Inverse relationship between the culture growth rate and the biosynthesis of heliomycin in the media with different sugars was shown. Arabinose and glucose provided the highest culture growth rate, while the levels of the antibiotic biosynthesis were the lowest. With the use of lactose the culture growth rate was the lowest and the antibiotic biosynthesis level was the highest. The level of the antibiotic biosynthesis was determined not only by the culture growth rate but also by the nature of the carbon source. With the use of glucose repression of heliomycin biosynthesis was much lower than that with the use of arabinose, while the culture growth rates were similar. Addition of glucose to the medium with lactose resulted in increasing of the antibiotic biosynthesis as long as the glucose concentration in the medium increased. Glycerol proved to be the most favourable carbon source for the biosynthesis of heliomycin. It also provided a comparatively high rate of the culture growth. Arabinose inhibited the antibiotic synthesis in the medium with glycerol and the inhibition level rose with an increase in the arabinose concentration. It is suggested that regulation of heliomycin biosynthesis is of the "glucose effect" type.


Recognition of potentially novel human disease-associated pathogens by implementation of systematic 16S rRNA gene sequencing in the diagnostic laboratory.
Clinical isolates that are difficult to identify by conventional means form a valuable source of novel human pathogens. We report on a 5-year study based on systematic 16S rRNA gene sequence analysis. We found 60 previously unknown 16S rRNA sequences corresponding to potentially novel bacterial taxa. For 30 of 60 isolates, clinical relevance was evaluated; 18 of the 30 isolates analyzed were considered to be associated with human disease.


A biomechanical switch regulates the transition towards homeostasis in oesophageal epithelium.
Epithelial cells rapidly adapt their behaviour in response to increasing tissue demands. However, the processes that finely control these cell decisions remain largely unknown. The postnatal period covering the transition between early tissue expansion and the establishment of adult homeostasis provides a convenient model with which to explore this question. Here, we demonstrate that the onset of homeostasis in the epithelium of the mouse oesophagus is guided by the progressive build-up of mechanical strain at the organ level. Single-cell RNA sequencing and whole-organ stretching experiments revealed that the mechanical stress experienced by the growing oesophagus triggers the emergence of a bright Krüppel-like factor 4 (KLF4) committed basal population, which balances cell proliferation and marks the transition towards homeostasis in a yes-associated protein (YAP)-dependent manner. Our results point to a simple mechanism whereby mechanical changes experienced at the whole-tissue level are integrated with those sensed at the cellular level to control epithelial cell fate.


Clinico-pathological and molecular subtypes of male breast carcinoma according to immunohistochemistry.
Male breast carcinoma is a rare condition, but with a trend of increase frequency. In our study, we investigate the clinico-pathological features and overall survival at 35 male cases of primary invasive breast carcinoma correlated with molecular subtypes defined by immunohistochemical profile. Based on immunohistochemical expression profiles of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor 2 (HER2) and Ki67, EGFR and CK5/6, the male breast cancers were classified into the following molecular subtypes: Luminal A, Luminal B, HER2+, triple negative and unclassified. In our study, we identified 65.7% as Luminal A subtype and 28.6% as Luminal B subtype. The difference was represented by two (5.7%) cases of triple negative subtype, but due to low number of patients, no correlations or prognostic significance could be assessed in these cases. No HER2 or unclassified subtypes were identified. Luminal A tumors are the most frequent subtype in MBC, with a better outcome than Luminal B subtype. We recorded high levels of ER and PR expression, which predict a better response to adjuvant hormonal therapy. At the time of diagnosis, most of the patients were aged and with an advance clinical stage, this requiring implementation of screening programs and increase education of population in order to an early detection.


Biomarkers of aging in women and the rate of longitudinal changes.
The purposes of this study were (1) to estimate biological age score (BAS) in Japanese healthy women based on the 4-7 years longitudinal data for physiological, hematological and biochemical examinations and (2) to examine the rate of aging changes in adult women based on the estimated BAS. The samples consisted of cross-sectional (n=981) and longitudinal (n=110) groups. Out of 31 variables examined, five variables (forced expiratory volume in 1.0 s, systolic blood pressure, mean corpuscular hemoglobin, glucose, albumin/globulin ratio) that met the following criteria: 1) significant cross-sectional correlation with age; 2) significant longitudinal change in the same direction as the cross-sectional correlation; and (3) assessment of redundancy, were selected as candidate biomarkers of aging. This variable set was then submitted into a principal component analysis, and the first principal component obtained from this analysis was used as an equation for assessing one's BAS. Individual BAS showed a high longitudinal stability of age-related changes, suggesting high predictive validity of our newly developed aging measurement equation. However, changes in the aging rate based on the estimated BAS were not constant. The mean slopes of the regression lines of BAS for the three age groups (age<45, 45</=age<65 yrs, 65</=age) were 0.095, 0.065, 0.138, respectively. One-way analysis of variance detected a significant difference (F=5.14, p<0.01) among the three age groups. These results suggest that the rate of aging in adult women is relatively slower until 65 years of age, but after 65, the rate of aging shows a rapid increase. We concluded that the longitudinal method used for selection of variables to compute the BAS was useful and theoretically valid compared to those obtained from cross-sectional data analysis.


Benzoyl peroxide microsphere cream as monotherapy and combination treatment of acne.
This office-based case series describes the use of a new formulation of benzoyl peroxide (BPO) delivered via a porous microsphere cream vehicle in patients with mild to moderate acne vulgaris. While BPO has been used for many years as antimicrobial in the treatment of acne, in recent years its use has been increasingly encouraged as part of a strategy designed to reduce Propionibacterium acnes (P acnes) resistance to antibiotics. Historically, a major drawback to BPO treatment has been irritation, which may be concentration-dependent or vehicle-dependent. A new BPO microsphere cream formulation has recently been introduced and appears to offer favorable efficacy with a very low potential for irritation. This article presents a series of patients from 2 private dermatologic practices treated with a new BPO microsphere cream formulation both as monotherapy and in combination with other acne drugs.


Dendritic Cell-Based In Situ Nanovaccine for Reprogramming Lipid Metabolism to Boost Tumor Immunotherapy.
Cancer vaccines have been considered to be an alternative therapeutic strategy for tumor therapy in the past decade. However, the popularity and efficacy of cancer vaccines were hampered by tumor antigen heterogeneity and the impaired function of cross-presentation in the tumor-infiltrating dendritic cells (TIDCs). To overcome these challenges, we engineered an in situ nanovaccine (named as TPOP) based on lipid metabolism-regulating and innate immune-stimulated nanoparticles. TPOP could capture tumor antigens and induce specific recognition by TIDCs to be taken up. Meanwhile, TPOP could manipulate TIDC lipid metabolism and inhibit de novo synthesis of fatty acids, thus improving the ability of TIDCs to cross-present by reducing their lipid accumulation. Significantly, intratumoral injection of TPOP combined with pretreatment with doxorubicin showed a considerable therapeutic effect in the subcutaneous mouse colorectal cancer model and melanoma model. Moreover, in combination with immune checkpoint inhibitors, such TPOP could markedly inhibit the growth of distant tumors by systemic antitumor immune responses. This work provides a safe and promising strategy for improving the function of immune cells by manipulating their metabolism and activating the immune system effectively for in situ cancer vaccines.


Molecular-genetic imaging: a nuclear medicine-based perspective.
Molecular imaging is a relatively new discipline, which developed over the past decade, initially driven by in situ reporter imaging technology. Noninvasive in vivo molecular-genetic imaging developed more recently and is based on nuclear (positron emission tomography [PET], gamma camera, autoradiography) imaging as well as magnetic resonance (MR) and in vivo optical imaging. Molecular-genetic imaging has its roots in both molecular biology and cell biology, as well as in new imaging technologies. The focus of this presentation will be nuclear-based molecular-genetic imaging, but it will comment on the value and utility of combining different imaging modalities. Nuclear-based molecular imaging can be viewed in terms of three different imaging strategies: (1) "indirect" reporter gene imaging; (2) "direct" imaging of endogenous molecules; or (3) "surrogate" or "bio-marker" imaging. Examples of each imaging strategy will be presented and discussed. The rapid growth of in vivo molecular imaging is due to the established base of in vivo imaging technologies, the established programs in molecular and cell biology, and the convergence of these disciplines. The development of versatile and sensitive assays that do not require tissue samples will be of considerable value for monitoring molecular-genetic and cellular processes in animal models of human disease, as well as for studies in human subjects in the future. Noninvasive imaging of molecular-genetic and cellular processes will complement established ex vivo molecular-biological assays that require tissue sampling, and will provide a spatial as well as a temporal dimension to our understanding of various diseases and disease processes.


SARS-CoV-2 infection and coronavirus disease 2019 severity in persons with HIV on antiretroviral treatment.
The relative susceptibility of people with HIV (PWH) to Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is debated. Numerous studies have been published with apparently contradictory findings, but comparisons are difficult because they have been conducted in populations with different characteristics (e.g. age, prevalence comorbidities) and have used different comparison groups (e.g. HIV-negative cohorts, coronavirus disease 2019 (COVID-19) hospitalized patients, general population), and because of challenges to measure the most important confounders. Here, we review the evidence regarding risk and severity of SARS-CoV-2 infection in PWH compared with persons without HIV. Publications originate largely from high-income settings where the majority of the PWH are on antiretroviral therapy (ART). Despite early evidence supporting higher frequency of SARS-CoV-2 testing in PWH on ART, HIV infection is not associated with SARS-CoV-2 infection, once confounding by socioeconomic characteristic is taken into account. Most publications identify increased COVID-19 severity in PWH compared with people without HIV from the general population or compared with COVID-19 hospitalized patients. The only study with an adequate comparison group to reduce confounding, has not identified differences in COVID-19 disease severity by HIV. Publications consistently identify that COVID-19 severity in PWH is not homogeneous and increases with age and baseline comorbidities. As PWH have a higher prevalence of comorbidities than people without HIV, examining their respective contribution to poor health outcomes is not straight forward as comorbidities could mediate the effect of HIV on COVID-19 outcomes.


Recombinant urease and urease DNA of Coccidioides immitis elicit an immunoprotective response against coccidioidomycosis in mice.
Coccidioides immitis antigens which stimulate a T helper cell 1 (Th1) pathway of host immune response are considered to be essential components of a vaccine against coccidioidomycosis. Recombinant urease (rURE) and recombinant heat shock protein 60 (rHSP60) of C. immitis were expressed in Escherichia coli and tested as vaccine candidates in BALB/c mice. A synthetic oligodeoxynucleotide which contained unmethylated CpG dinucleotides and was previously shown to enhance a murine Th1 response was used as an immunoadjuvant. T cells isolated from the spleens and lymph nodes of the rURE- and rHSP60-immune mice showed in vitro proliferative responses to the respective recombinant protein, but only those T lymphocytes from rURE-immunized mice revealed markedly elevated levels of expression of selected Th1-type cytokine genes. BALB/c mice immunized subcutaneously with rURE and subsequently challenged by the intraperitoneal (i.p.) route with a lethal inoculum of C. immitis arthroconidia demonstrated a significant reduction in the level of C. immitis infection compared to control animals. rHSP60 was much less effective as a protective antigen. Evaluation of cytokine gene expression in lung tissue and levels of recombinant urease-specific immunoglobulins (immunoglobulin G1 [IgG1] versus IgG2a) in murine sera at 12 days after challenge provided additional evidence that immunization with rURE stimulated a Th1 response to the pathogen. Urease was further evaluated by expression of the URE gene in a mammalian plasmid vector (pSecTag2A.URE) which was used to immunize mice by the intradermal route. In this case, 82% of the vector construct-immunized animals survived more than 40 days after i.p. infection, compared to only 10% of the mice immunized with the vector alone. In addition, 87% of the pSecTag2A.URE-immunized survivors had sterile lungs and spleens. These data support the need for further evaluation of the C. immitis urease as a candidate vaccine against coccidioidomycosis.


Profiling cellular protein complexes by proximity ligation with dual tag microarray readout.
Patterns of protein interactions provide important insights in basic biology, and their analysis plays an increasing role in drug development and diagnostics of disease. We have established a scalable technique to compare two biological samples for the levels of all pairwise interactions among a set of targeted protein molecules. The technique is a combination of the proximity ligation assay with readout via dual tag microarrays. In the proximity ligation assay protein identities are encoded as DNA sequences by attaching DNA oligonucleotides to antibodies directed against the proteins of interest. Upon binding by pairs of antibodies to proteins present in the same molecular complexes, ligation reactions give rise to reporter DNA molecules that contain the combined sequence information from the two DNA strands. The ligation reactions also serve to incorporate a sample barcode in the reporter molecules to allow for direct comparison between pairs of samples. The samples are evaluated using a dual tag microarray where information is decoded, revealing which pairs of tags that have become joined. As a proof-of-concept we demonstrate that this approach can be used to detect a set of five proteins and their pairwise interactions both in cellular lysates and in fixed tissue culture cells. This paper provides a general strategy to analyze the extent of any pairwise interactions in large sets of molecules by decoding reporter DNA strands that identify the interacting molecules.


Transcriptional control of early tract formation in the embryonic chick midbrain.
The earliest step in establishing the complex neuronal networks in the vertebrate brain is the formation of a scaffold of axon tracts. How the formation of the early axon scaffold is controlled at the molecular level is unclear. Forming part of the scaffold, neurons located at the ventral midbrain-forebrain border (MFB) give rise to the medial longitudinal fascicle (mlf) and the posterior commissure (pc). We demonstrate that the homeobox genes Sax1, Six3, Emx2 and Pax6 are expressed in distinct domains in this area, suggesting that the specification of mlf and pc neurons might be controlled by the combinatorial activity of these transcription factors. We have tested this hypothesis by analysing the function of Sax1 in the embryonic chick brain. Gain-of-function experiments with Sax1 result in alterations to the early axon scaffold, most prominently an enlargement of the mlf at the expense of the pc. Ectopic expression of Sax1 also affects the expression of other ventral homeobox genes, particularly Six3 and Emx2. Our results indicate that the specification of neurons forming the early axon scaffold is governed by a homeobox code, thus resembling the mechanism of neuronal specification in the spinal cord.


'Too late or too soon': The ethics of advance care planning in dementia setting.
Advance care planning (ACP) is considered a pivotal aid in the decision-making process, especially for many people living with dementia, who inevitably will lose the capacity to make decisions at the end of life. In Italy, ACP has been recently regulated by law 219/2017, leading to the investigation of how physicians deal with ACP in dementia. In order to comprehend the perception of physicians who provide care for patients with dementia regarding ACP and to describe their difficulties and needs, this study presents the results of a qualitative research study performed within a local health authority in northern Italy. Data were collected by semi-structured interviews with 11 physicians dealing with patients with dementia and were analysed using interpretative phenomenological analysis. Four superordinate themes were identified, describing both the difficulties and challenges perceived by physicians in performing shared ACP in their daily activities. This study demonstrates that physicians experience troubles synchronizing the time of shared ACP with the more rapid development of dementia. Results also suggest that further training about communication on ACP is urgently required to ease the implementation of shared ACP. A broader involvement of institutions and other professionals is also important in spreading ACP among healthcare institutions.


Quantitative allele-specific expression and DNA methylation analysis of H19, IGF2 and IGF2R in the human placenta across gestation reveals H19 imprinting plasticity.
Imprinted genes play important roles in placental differentiation, growth and function, with profound effects on fetal development. In humans, H19 and IGF2 are imprinted, but imprinting of IGF2R remains controversial. The H19 non-coding RNA is a negative regulator of placental growth and altered placental imprinting of H19-IGF2 has been associated with pregnancy complications such as preeclampsia, which have been attributed to abnormal first trimester placentation. This suggests that changes in imprinting during the first trimester may precede aberrant placental morphogenesis. To better understand imprinting in the human placenta during early gestation, we quantified allele-specific expression for H19, IGF2 and IGF2R in first trimester (6-12 weeks gestation) and term placentae (37-42 weeks gestation) using pyrosequencing. Expression of IGF2R was biallelic, with a mean expression ratio of 49:51 (SD = 0.07), making transient imprinting unlikely. Expression from the repressed H19 alleles ranged from 1-25% and was higher (P<0.001) in first trimester (13.5 ± 8.2%) compared to term (3.4 ± 2.1%) placentae. Surprisingly, despite the known co-regulation of H19 and IGF2, little variation in expression of the repressed IGF2 alleles was observed (2.7 ± 2.0%). To identify regulatory regions that may be responsible for variation in H19 allelic expression, we quantified DNA methylation in the H19-IGF2 imprinting control region and H19 transcription start site (TSS). Unexpectedly, we found positive correlations (P<0.01) between DNA methylation levels and expression of the repressed H19 allele at 5 CpG's 2000 bp upstream of the H19 TSS. Additionally, DNA methylation was significantly higher (P<0.05) in first trimester compared with term placentae at 5 CpG's 39-523 bp upstream of the TSS, but was not correlated with H19 repressed allele expression. Our data suggest that variation in H19 imprinting may contribute to early programming of placental phenotype and illustrate the need for quantitative and robust methodologies to further elucidate the role of imprinted genes in normal and pathological placental development.


Systematization method for distinguishing plastic groups by using NIR spectroscopy.
A systematic classification method for polymers is not yet available in case of using near infrared spectra (NIR). That is why we have been searching for a systematic method. Because raw NIR spectra usually have few obvious peaks, NIR spectra have been pretreated by 2nd derivation for taking well modulated spectra. After the pretreatment, we applied classification and regression trees (CART) to the discrimination between the spectra and the species of polymers. As a result, we obtained a relatively simple classification tree. Judging from the obtained splitting conditions and the classified polymers, we concluded that obtained knowledge on the chemical function groups estimated by the important wavelength regions is not always applicable to this classification tree. However, we clarified the splitting rules for polymer species from the NIR spectral point of view.


The reaction of ornithine aminotransferase with ornithine.
Rat liver ornithine aminotransferase is found to exchange the pro-S hydrogen on the delta-carbon atom of ornithine exclusively, thus showing that the mammalian enzyme transfers the delta-amino group and not the alpha-amino group as has been demonstrated with the plant enzyme [Mestichelli, Gupta & Spenser (1979) J. Biol. Chem. 254, 640-647]. The enzyme also transfers the alpha-amino group of glutamate and the kinetics of the half reactions between the enzyme and both amino acids are compared. Rate and dissociation constants for both reactions are determined.


Membrane permeability induced by hepatitis A virus proteins 2B and 2BC and proteolytic processing of HAV 2BC.
The ability to rearrange membranes is a unique feature of nonstructural proteins 2B, 2C, and 2BC of some picornaviruses. To analyze in detail membrane binding of the respective proteins of hepatitis A virus (HAV), they were transiently expressed in the vaccinia/T7 system, and their effect on membrane permeability was studied using beta-galactosidase as reporter. Although 2C had no effect, the significantly increased reporter activity observed in the extracellular space of 2B- and 2BC-expressing cells points to a specific effect of HAV proteins 2B and 2BC on membrane permeability. In biochemical fractionation studies, HAV 2C and 2BC showed properties of intregral membrane proteins, whereas 2B was associated with membranes as a peripheral protein. Proteinase 3C-mediated cleavage of precursor 2BC in vivo was most efficient when the enzyme was coexpressed in its precursor forms P3 or 3ABC, which both include the membrane-anchoring domain 3A. 3ABC showed the same solubility pattern as 2BC, suggesting that colocalization of 2BC and 3ABC might be required for the efficient liberation of 2B and 2C and occurs on membranes that have been proposed as the site of viral RNA replication.


On-road measurement of vehicle tailpipe emissions using a portable instrument.
A study design procedure was developed and demonstrated for the deployment of portable onboard tailpipe emissions measurement systems for selected highway vehicles fueled by gasoline and E85 (a blend of 85% ethanol and 15% gasoline). Data collection, screening, processing, and analysis protocols were developed to assure data quality and to provide insights regarding quantification of real-world intravehicle variability in hot-stabilized emissions. Onboard systems provide representative real-world emissions measurements; however, onboard field studies are challenged by the observable but uncontrollable nature of traffic flow and ambient conditions. By characterizing intravehicle variability based on repeated data collection runs with the same driver/vehicle/route combinations, this study establishes the ability to develop stable modal emissions rates for idle, acceleration, cruise, and deceleration even in the face of uncontrollable external factors. For example, a consistent finding is that average emissions during acceleration are typically 5 times greater than during idle for hydrocarbons and carbon dioxide and 10 times greater for nitric oxide and carbon monoxide. A statistical method for comparing on-road emissions of different drivers is presented. Onboard data demonstrate the importance of accounting for the episodic nature of real-world emissions to help develop appropriate traffic and air quality management strategies.


Superoxide-producing lipoprotein fraction from Stevia leaves: definition of specific activity.
Stevia rebaudiana Bertoni has various pharmacological actions, which includes antidiabetic, antioxidant, anti-inflammatory activities. The superoxide and consequently NADPH oxidase (Nox) are relevant targets involved in biological effects of Stevia. The presence of NADPH-containing superoxide-producing lipoprotein (suprol) in Stevia leaves has not yet been tested. The mechanism of producing superoxide radicals (O2-) by suprol was determined in vitro, which is associated with the electron transfer from NADPH in the composition of suprol by traces of transition metal ions (Fe3+ or Cu2+) to molecular oxygen, turning it into O2-. It is expected that the therapeutic efficacy of Stevia leaves is caused by specific activity of superoxide-producing lipoprotein fraction. For the first time, from the dry leaves of Stevia the NADPH-containing superoxide-producing lipoprotein was isolated and purified. The specific content of suprol (milligrams in 1 g of Stevia leaves- mg/g) was determined after desalination of suprol and lyophilization. According to the method provided, the specific content of the isolated suprol from Stevia's leaves was up to 4.5 ± 0.2 mg / g (yields up to 68.5 ± 4.7%, p < 0.05, n = 6). Nox forms a stable complex with suprol. The optical absorption spectrum of the Nox-suprol complex represents the overlapping suprol and Nox spectra, with a certain background increase and characteristic features of optical absorption for Nox. Due to O2- producing capacity suprol-Nox complex discolors KMnO4 solutions, Coomassie brilliant blue, restores nitrotetrazolium blue to formazan and oxidizes epinephrine to adrenochrome. The oxidation activity of adrenaline is 50.3 ± 5.1 U / mg / ml (p < 0.05, n = 6). Superoxide-producing lipoprotein fraction-Nox complex from Stevia leaves (membranes) can modulate redox regulated signaling pathways and may play a positive role in type-2 diabetes by means of adrenaline oxidation mechanism.


Delving into the social ecology of leisure-time physical activity among adolescents from south eastern Spain.
Worldwide, there is a growing concern with adolescents' low levels of physical activity (PA). We used a comprehensive social ecological framework to uncover factors associated with leisure-time physical activity (LTPA) among adolescents from southeastern Spain. A population-based sample of 3249 adolescents aged 12-17 participated in a school-based survey in 2006. Potential correlates of participation in and level of LTPA were assessed through self-report. LTPA levels were also self-reported. We used gender-stratified logistic regression models to examine the associations among the variables of interest. Consistent with a social ecological perspective, analyses revealed several factors, corresponding to different levels of organization (demographic, biological, psychological, behavioral, social) and behavioral settings (family, peer group, school), significantly associated with LTPA. Some of these factors varied as a function of gender and depending on whether the outcome considered was nonparticipation vs. participation in LTPA or high vs. low level of involvement among participants. Overall, the findings highlight the role of health-related participation motives, significant others' attitudes toward PA, and grade in physical education as correlates of LTPA in this sample. Continued research is necessary to understand the complex interplay of factors and settings associated with adolescent LTPA and the role of gender.


New records of amphipod crustaceans along the Israeli Mediterranean coast, including a rare Mediterranean endemic species, Maera schieckei Karaman & Ruffo, 1971.
A survey has been carried out at four Israeli rocky sites to evaluate the diversity of the amphipod fauna on various hard substrates, still scarcely monitored, as potential pabulum for amphipod crustacean species. A survey of shallow rocky reefs along the Mediterranean coast of Israel recovered 28 species and integrated the Amphipoda checklist for the country ofIsrael with 12 newly-recorded species. Such renewed national list includes Maera schieckei Karaman & Ruffo, 1971, a rare species endemic to the Mediterranean Sea, recorded here for the first time from the southern Levant Basin. The species, described from specimens collected in the Tyrrhenian Sea in 1970, has been only recorded eight times within the whole Mediterranean Sea. A revision of the bibliography on the distribution and ecology of M. schieckei showed that, although mentioned only for the western Mediterranean basin by some authors, it is listed in the checklist of amphipods of the Aegean Sea and neighbouring seas and has been found in the eastern Mediterranean basin since 1978. Maera schieckei was rarely found in the Mediterranean, one of the most studied marine biogeographic region as concerns the amphipod fauna; and the species seems to prefer bays or gulf areas. The role of updating and monitoring faunal composition should be re-evaluated.


Apparent cortisone reductase deficiency: a functional defect in 11beta-hydroxysteroid dehydrogenase type 1.
A 36-yr-old woman was referred to the endocrine clinic for investigation of oligomenorrhea, hirsutism, and acne. She was plethoric and overweight with central fat distribution. Plasma cortisol was normal, but her adrenal glands were enlarged (CT scan). Urinary tetrahydrocortisone excretion rate was consistently high, raising the possibility of 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) deficiency. In addition, 5beta- reduction of cortisol and cortisone was markedly enhanced. The levels of all cortisol metabolites were suppressed normally with dexamethasone, but conversion of oral cortisone acetate to plasma cortisol was delayed and subnormal compared with that of healthy volunteers. This was accompanied by a larger than normal increase in plasma cortisone concentration. Thus, the defect appears to be in 11beta-HSD1 activity and not in 5beta-reductase activity. Three close relatives of the subject showed no comparable abnormalities, and analysis of the coding region and exon/intron boundaries of the 11beta-HSD1 gene of the case revealed no differences from the consensus sequence. The defect may lie outside the coding region. Alternatively, some other inherited or acquired defect may lead to inhibition of this enzyme system.


The impact of saliva collection methods on measured salivary biomarker levels.
Saliva diagnostics have become increasingly popular due to their non-invasive nature and patient-friendly collection process. Various collection methods are available, yet these are not always well standardized for either quantitative or qualitative analysis. In line, the objective of this study was to evaluate if measured levels of various biomarkers in the saliva of healthy individuals were affected by three distinct saliva collection methods: 1) unstimulated saliva, 2) chew stimulated saliva, and 3) oral rinse. Saliva samples from 30 healthy individuals were obtained by the three collection methods. Then, the levels of various salivary biomarkers such as proteins and ions were determined. It was found that levels of various biomarkers obtained from unstimulated saliva were comparable to those in chew stimulated saliva. The levels of potassium, sodium, and amylase activity differed significantly among the three collection methods. Levels of all biomarkers measured using the oral rinse method significantly differed from those obtained from unstimulated and chew-stimulated saliva. In conclusion, both unstimulated and chew-stimulated saliva provided comparable levels for a diverse group of biomarkers. However, the results obtained from the oral rinse method significantly differed from those of unstimulated and chew-stimulated saliva, due to the diluted nature of the saliva extract.


Role of mast cells in intestinal mucosal function: studies in models of hypersensitivity and stress.
A single layer of epithelial cells lines the gastrointestinal tract, forming a critical barrier between the lunminal contents, which includes antigens and other noxious substances, and the body proper. It has become clear in recent years that the role of mast cells in the gastrointestinal mucosa is not only to react to antigens, but also to actively regulate the barrier and transport properties of the intestinal epithelium. Mucosal mast cells respond to both IgE/antigen-dependent and non-IgE-dependent stimulation, releasing bioactive mediators into adjacent tissues where they induce physiological responses. Studies in models of hypersensitivity and stress have provided evidence that changes in mucosal function are due to either direct action of mast cell mediators on epithelial receptors and/or indirect action via nerves/neurotransmitters.


Biosysthesis of corn starch palmitate by lipase Novozym 435.
Esterification of starch was carried out to expand the usefulness of starch for a myriad of industrial applications. Lipase B from Candida antarctica, immobilized on macroporous acrylic resin (Novozym 435), was used for starch esterification in two reaction systems: micro-solvent system and solvent-free system. The esterification of corn starch with palmitic acid in the solvent-free system and micro-solvent system gave a degree of substitution (DS) of 1.04 and 0.0072 respectively. Esterification of corn starch with palmitic acid was confirmed by UV spectroscopy and IR spectroscopy. The results of emulsifying property analysis showed that the starch palmitate with higher DS contributes to the higher emulsifying property (67.6%) and emulsion stability (79.6%) than the native starch (5.3% and 3.9%). Modified starch obtained by esterification that possesses emulsifying properties and has long chain fatty acids, like palmitic acid, has been widely used in the food, pharmaceutical and biomedical applications industries.


The Mini-Addenbrooke's Cognitive Examination (M-ACE) as a brief cognitive screening instrument in Mild Cognitive Impairment and mild Alzheimer's disease.
The Mini-Addenbrooke's Cognitive Examination (M-ACE) is a brief cognitive screening test that evaluates four main cognitive domains (orientation, memory, language and visuospatial function) with a maximum score of 30 points and administration time of five minutes. To assess the performance of healthy elderly, MCI patients and mild AD patients using the Brazilian version of the M-ACE. The test was applied to a group of 36 Mild Cognitive Impairment (MCI), 23 mild Alzheimer's Disease (AD) and 25 cognitive healthy elderly. All participants were aged ≥60 years. The M-ACE displayed high internal consistency (Cronbach alpha >0.8; 95% CI 0.7-0.8) and proved effective for differentiating the AD group from MCI and control groups, providing superior accuracy than the MMSE (the cut-off point of 20 points had the highest sensitivity and specificity - 95.6% and 90.16% respectively, with a high area under the curve - AUC=0.8; 95% CI 0.7-0.9). Performance on the M-ACE was strongly correlated with that of the MMSE and Functional Activities Questionnaire (FAQ). The M-ACE was not accurate in discriminating MCI from control subjects. The M-ACE is a brief screening test which provided high accuracy for diagnosing AD in this sample. The suggested cut-off point in this study was 20 points for AD.


Promotion of glioma cell survival by acyl-CoA synthetase 5 under extracellular acidosis conditions.
Extracellular acidosis (low pH) is a tumor microenvironmental stressor that has a critical function in the malignant progression and metastatic dissemination of tumors. To survive under stress conditions, tumor cells must evolve resistance to stress-induced toxicity. Acyl-CoA synthetase 5 (ACSL5) is a member of the ACS family, which converts fatty acid to acyl-CoA. ACSL5 is frequently overexpressed in malignant glioma, whereas its functional significance is still unknown. Using retrovirus-mediated stable gene transfer (gain of function) and small interfering RNA-mediated gene silencing (loss of function), we show here that ACSL5 selectively promotes human glioma cell survival under extracellular acidosis. ACSL5 enhanced cell survival through its ACS catalytic activity. To clarify the genome-wide changes in cell signaling pathways by ACSL5, we performed cDNA microarray analysis and identified an ACSL5-dependent gene expression signature. The analysis revealed that ACSL5 was critical to the expression of tumor-related factors including midkine (MDK), a heparin-binding growth factor frequently overexpressed in cancer. Knockdown of MDK expression significantly attenuated ACSL5-mediated survival under acidic state. These results indicate that ACSL5 is a critical factor for survival of glioma cells under acidic tumor microenvironment, thus providing novel molecular basis for cancer therapy.


Champion-level drone racing using deep reinforcement learning.
First-person view (FPV) drone racing is a televised sport in which professional competitors pilot high-speed aircraft through a 3D circuit. Each pilot sees the environment from the perspective of their drone by means of video streamed from an onboard camera. Reaching the level of professional pilots with an autonomous drone is challenging because the robot needs to fly at its physical limits while estimating its speed and location in the circuit exclusively from onboard sensors1. Here we introduce Swift, an autonomous system that can race physical vehicles at the level of the human world champions. The system combines deep reinforcement learning (RL) in simulation with data collected in the physical world. Swift competed against three human champions, including the world champions of two international leagues, in real-world head-to-head races. Swift won several races against each of the human champions and demonstrated the fastest recorded race time. This work represents a milestone for mobile robotics and machine intelligence2, which may inspire the deployment of hybrid learning-based solutions in other physical systems.


Minocycline Reduces the Severity of Autonomic Dysreflexia after Experimental Spinal Cord Injury.
Spinal cord injury (SCI) is a devastating neurological condition for which there is no effective treatment to restore neurological function. The development of new treatments for those with SCI may be hampered by the insensitivity of clinical tools to assess motor function in humans. Treatments aimed at preserving neuronal function through anti-inflammatory pathways (i.e., neuroprotection) have been a mainstay of pre-clinical SCI research for decades. Minocycline, a clinically available antibiotic agent with anti-inflammatory properties, has demonstrated promising neuroprotective effects in a variety of animal models and improved motor recovery in a Phase-2 human trial. Here, we leveraged our recently developed T3 severe contusion model in the rat to determine the ability of minocycline to preserve descending sympathoexcitatory axons and improve cardiovascular control after SCI. Forty-one male Wistar rats were randomized to either a treatment group (minocycline; n = 20) or a control group (vehicle; n = 21). All rats received a severe T3 contusion. Minocycline (or vehicle) was administered intraperitoneally at one hour post-injury (90 mg/kg), then every 12 h for two weeks (45 mg/kg). Neuroanatomical correlates (lesion area, descending sympathoexcitatory axons) were assessed, in addition to an assessment of cardiovascular control (hemodynamics, autonomic dysreflexia) and motor behavior. Here, we show that minocycline reduces lesion area, increases the number of descending sympathoexctitatory axons traversing the injury site, and ultimately reduces the severity of autonomic dysreflexia. Finally, we show that autonomic dysreflexia is a more sensitive marker of treatment stratification than motor function.


Accurate predictions of population-level changes in sequence and structural properties of HIV-1 Env using a volatility-controlled diffusion model.
The envelope glycoproteins (Envs) of HIV-1 continuously evolve in the host by random mutations and recombination events. The resulting diversity of Env variants circulating in the population and their continuing diversification process limit the efficacy of AIDS vaccines. We examined the historic changes in Env sequence and structural features (measured by integrity of epitopes on the Env trimer) in a geographically defined population in the United States. As expected, many Env features were relatively conserved during the 1980s. From this state, some features diversified whereas others remained conserved across the years. We sought to identify "clues" to predict the observed historic diversification patterns. Comparison of viruses that cocirculate in patients at any given time revealed that each feature of Env (sequence or structural) exists at a defined level of variance. The in-host variance of each feature is highly conserved among individuals but can vary between different HIV-1 clades. We designate this property "volatility" and apply it to model evolution of features as a linear diffusion process that progresses with increasing genetic distance. Volatilities of different features are highly correlated with their divergence in longitudinally monitored patients. Volatilities of features also correlate highly with their population-level diversification. Using volatility indices measured from a small number of patient samples, we accurately predict the population diversity that developed for each feature over the course of 30 years. Amino acid variants that evolved at key antigenic sites are also predicted well. Therefore, small "fluctuations" in feature values measured in isolated patient samples accurately describe their potential for population-level diversification. These tools will likely contribute to the design of population-targeted AIDS vaccines by effectively capturing the diversity of currently circulating strains and addressing properties of variants expected to appear in the future.


Influence of power supply in the feasibility of Phaeodactylum tricornutum cultures.
The influence of fluid-dynamic conditions on the yield of Phaeodactylum tricornutum microalgal cultures was analyzed in two stages: first, the influence of air flow rate; second, the influence of using fluid-moving pumps for recirculating the culture. With respect to the air flow rate, the yield of the cultures increased with the aeration rate up to values of 2.0 v/v/min, then stress was observed and the yield of the cultures decreased. With respect to the influence of mechanical power supply for liquid impulsion, three different types of pumps--centrifugal, pulse, and peristaltic--were essayed at different power supplies. The cultures were stressed for the three types of pumps essayed. For each pump, the higher the power supply the lower was the Fv/Fm value and the higher was the stress at which cells were exposed. The highest measured stress was when the culture was moved with the centrifugal pump. Despite measured stress, for all the experiments stable steady states were reached, thus indicating that cells reduced their yield but did not die, as was verified by cell viability measurements. It was observed that the increase of the power supply improved the frequency of light exposition thus enhancing the yield of the cultures. However, the higher the power supply, the lower the microeddy length scale; therefore, stress could appear. Data demonstrated that the microeddy length scale was always much higher than cell size and therefore the turbulence was not responsible for stress. Also, the mass transfer was discarded as responsible for yield reduction. It was concluded that the shear rate was the factor determining the existence of stress phenomena. The evaluation of these shear rates demonstrated that values above 30-80 s(-1) damaged the cells strongly. These data were verified in an outdoor pilot-scale tubular photobioreactor that was implemented with the same type of pumps, thus demonstrating the necessity to take into account this factor in the design and scale-up of microalgal photobioreactors.


Color discrimination with broadband photoreceptors.
Color vision is commonly assumed to rely on photoreceptors tuned to narrow spectral ranges. In the ommatidium of Drosophila, the four types of so-called inner photoreceptors express different narrow-band opsins. In contrast, the outer photoreceptors have a broadband spectral sensitivity and were thought to exclusively mediate achromatic vision. Using computational models and behavioral experiments, we demonstrate that the broadband outer photoreceptors contribute to color vision in Drosophila. The model of opponent processing that includes the opsin of the outer photoreceptors scored the best fit to wavelength discrimination data. To experimentally uncover the contribution of individual photoreceptor types, we restored phototransduction of targeted photoreceptor combinations in a blind mutant. Dichromatic flies with only broadband photoreceptors and one additional receptor type can discriminate different colors, indicating the existence of a specific output comparison of the outer and inner photoreceptors. Furthermore, blocking interneurons postsynaptic to the outer photoreceptors specifically impaired color but not intensity discrimination. Our findings show that receptors with a complex and broad spectral sensitivity can contribute to color vision and reveal that chromatic and achromatic circuits in the fly share common photoreceptors.


Increased bleomycin-induced skin fibrosis in mice lacking the Th1-specific transcription factor T-bet.
Fibrosis, the pathological hallmark of scleroderma and related conditions, is due to sustained activation of tissue fibroblasts. Accumulating evidence implicates cytokine networks in initiating, and propagating or terminating fibroblast activation, and the specific cytokine phenotype dictates evolution of the fibrotic response toward either resolution or scarring. In particular, cytokines that promote fibroblast proliferation and myofibroblast differentiation and extracellular matrix (ECM) accumulation functionally define a type 2 (Th2) immune response, whereas interferon-gamma, which suppresses diverse fibroblast activities, defines a type 1 (Th1) immune response. It remains unclear what role the balance between Th1 and Th2 cytokines plays in the pathogenesis of fibrosis. Here we used bleomycin-induced skin fibrosis as a murine model for human scleroderma in order to study the fibrotic response in mice lacking T-bet, a transcription factor that is essential for initiating Th1 lineage development of CD4+ T lymphocytes. Spleen cells from T-bet null (T-bet(-/-)) mice exhibited a typical Th2 cytokine profile ex vivo, with elevated production of interleukin-4 (IL-4), IL-5 and IL-13, and diminished production of interferon-gamma. Bleomycin-induced early mast cells and eosinophil accumulation, and eosinophil degranulation, in the lesional tissue were greater in T-bet(-/-) mice than in wild-type control mice. At a later time point, T-bet(-/-) mice developed significantly more extensive dermal and especially hypodermal fibrosis. Elevated TGF-beta expression and intracellular Smad activation were prominent in lesional skin. Infiltrating eosinophils appeared to be an important cellular source of TGF-beta. These results demonstrate that in mice lacking T-bet bleomycin induced exaggerated skin fibrosis, suggesting that T-bet has an important physiologic role in regulation of tissue repair by promoting Th1 immune responses that prevent excessive ECM accumulation.


QT prolongation and serum sotalol concentration are highly correlated following intravenous and oral sotalol.
The aim of this study was to evaluate the correlation between QT interval (QT) and serum sotalol concentration following a single low dose of oral and intravenous sotalol. Fifteen healthy volunteers received 75 mg intravenous sotalol over 2.5 h and 80 mg oral sotalol in a random order. Serum sotalol concentrations and 12-lead electrocardiograms were obtained simultaneously at baseline and 7 times following dosings. Rate-corrected QT (QTc) was calculated by the Bazett, Fridericia and Framingham formulas. Linear regression analysis was performed between sotalol concentrations and QT measurements. Significant QT prolongation was seen at very low sotalol doses and serum concentrations. QTc intervals calculated by the Framingham and Fridericia formulas showed the strongest and virtually identical correlations with serum sotalol concentration (r >or= 0.97, p < 0.001) following oral and intravenous administrations. The equation QTc = 0.0342 (sotalol concentration) + 398 closely predicted actual QTc at any sotalol concentration. A strong correlation was observed between serum sotalol concentration and QTc prolongation across the entire concentration range. Low-dose sotalol caused significant QT prolongation. At similar concentrations, intravenous and oral sotalol caused similar QT and QTc effects. Knowing the QT effect can be used to guide further dose increase.


Dysfunction of homeostatic control of dopamine by astrocytes in the developing prefrontal cortex leads to cognitive impairments.
Astrocytes orchestrate neural development by powerfully coordinating synapse formation and function and, as such, may be critically involved in the pathogenesis of neurodevelopmental abnormalities and cognitive deficits commonly observed in psychiatric disorders. Here, we report the identification of a subset of cortical astrocytes that are competent for regulating dopamine (DA) homeostasis during postnatal development of the prefrontal cortex (PFC), allowing for optimal DA-mediated maturation of excitatory circuits. Such control of DA homeostasis occurs through the coordinated activity of astroglial vesicular monoamine transporter 2 (VMAT2) together with organic cation transporter 3 and monoamine oxidase type B, two key proteins for DA uptake and metabolism. Conditional deletion of VMAT2 in astrocytes postnatally produces loss of PFC DA homeostasis, leading to defective synaptic transmission and plasticity as well as impaired executive functions. Our findings show a novel role for PFC astrocytes in the DA modulation of cognitive performances with relevance to psychiatric disorders.


[The research protocol VI: How to choose the appropriate statistical test. Inferential statistics].
The statistical analysis can be divided in two main components: descriptive analysis and inferential analysis. An inference is to elaborate conclusions from the tests performed with the data obtained from a sample of a population. Statistical tests are used in order to establish the probability that a conclusion obtained from a sample is applicable to the population from which it was obtained. However, choosing the appropriate statistical test in general poses a challenge for novice researchers. To choose the statistical test it is necessary to take into account three aspects: the research design, the number of measurements and the scale of measurement of the variables. Statistical tests are divided into two sets, parametric and nonparametric. Parametric tests can only be used if the data show a normal distribution. Choosing the right statistical test will make it easier for readers to understand and apply the results.


Emotional processing in a salient motion context: integration of motion and emotion in both V5/hMT+ and the amygdala.
Recent studies have suggested that the extent to which primary task demands draw on attentional resources determines whether or not task-irrelevant emotional stimuli are processed. Another important factor that can bias task-relevant and task-irrelevant stimulus competition is the bottom-up factor of stimulus salience. Here, we investigated the effect of stimulus salience associated with a primary motion task on the processing of emotional face distractors. Faces of different emotional valences were presented within a context of randomly moving dots. Subjects had to detect short intervals of coherent motion while ignoring the background faces. Task salience was manipulated by the level of motion coherence of the dots with high motion coherence being associated with high salience. Using functional magnetic resonance imaging, we show that emotional faces, compared with neutral faces, more strongly interfered with the primary task, as reflected in significant signal decreases in task-related motion area V5/hMT+. In addition, these faces elicited significant signal increases in the left amygdala. Most importantly, task salience was found to further increase amygdala's activity when presented together with an emotional face. Our data support a more general role of the amygdala as a behavioral relevance detector, which flexibly integrates behavioral relevant, salient context information to decode the emotional content of a visual scene.


Patients' predispositions to discuss health issues affecting quality of life.
Many patients are reluctant to talk to family physicians about certain health issues that affect quality of life. This research examines factors that influence a patient's willingness to discuss three aspects of health-related quality of life: overall well-being, physical health, and psychosocial health. Prior to their consultations, 254 patients completed questionnaires that assessed their perceptions of health limitations, beliefs about the family physician's job responsibilities, willingness to discuss various topics with the physician, and relational history with the physician (ie, number of previous visits). On the post-visit instrument, patients reported whether certain topics were discussed during the consultation and their satisfaction with the physician. Patients were more willing to discuss all aspects of health-related quality of life when they believed it was the doctor's job to ask about these issues and when they had a longer relational history with the doctor. Patients' health limitations did not predispose patients to talk about certain issues. Patients were more satisfied with care when there was more discussion of the patient's overall well-being and physical health. However, discussions about psychosocial issues were not related to patient satisfaction. A theoretical model is proposed that may be useful in explaining why patients often vary in their willingness to discuss certain topics, particularly psychosocial issues. Future research should analyze how doctors and patients introduce, manage, and terminate discussions of health-related quality of life.


Isolation of multiple forms of DNA polymerase delta: evidence of proteolytic modification during isolation.
The subunit structures of a number of human placenta DNA polymerase delta preparations were investigated by Western blotting with polyclonal antisera and by activity staining following polyacrylamide gel electrophoresis. When immunoblots and activity stains were performed on different enzyme preparations, putative catalytic subunits of (a) 170, (b) 120, or (c) 50-70 kilodaltons (kDa) were observed. It was also observed that the lower molecular weight forms could be generated upon storage of the preparations. Western blotting of human placental tissue extracts showed that the major immunoreactive polypeptide was 160-170 kDa. Treatment of the extracts with trypsin or Staphylococcus aureus V8 protease led to the generation of immunoreactive polypeptides of lower molecular weights. These studies suggest that the 120-kDa and lower forms of the enzyme are generated via uncontrolled proteolysis and provide a rationale for the observation of different apparent subunit structures previously reported for DNA polymerase delta. In addition, these findings suggest that DNA polymerase delta has a catalytic domain which resides in a protease-resistant domain.


Pisum sativum Response to Nitrate as Affected by Rhizobium leguminosarum-Derived Signals.
Legumes are suitable for the development of sustainable agroecosystems because of their ability to use atmospheric N2 through symbiotic nitrogen fixation (SNF). However, a basic NO3- input is necessary before SNF takes place to ensure successful seedling establishment. Since Rhizobia not only induce nodulation but also affect root branching by stimulating the development of lateral roots, and NO3- as a signal also modulates root system architecture, we investigated whether Rhizobium-derived signals interfere in nitrate signaling. Here, we bring evidence that (i) Rhizobium-altered NO3--mediated processes in pea expressions of major players in NO3- transport, sensing, and signaling were affected, and (ii) the characteristic limitation of root foraging and branching in response to NO3- supply was abolished. The number of tertiary roots per secondary root was higher in infected compared to uninfected peas, thus indicating that the Rhizobium effect allows for favorable management of trade-offs between nodules growth for nitrogen capture and root foraging for water and other nutrient uptake in pea. The outcome of this basic research can be used to produce molecular tools for breeding pea genotypes able to develop deep-foraging and branched root systems, and more competitive architectures and molecular levels for soil NO3- absorption during seedling establishment without jeopardizing nodulation.