Fourteen-year oncological and functional outcomes of high-intensity focused ultrasound in localized prostate cancer.
WHAT'S KNOWN ON THE SUBJECT? AND WHAT DOES THE STUDY ADD?: High-intensity focused ultrasound (HIFU) is an alternative treatment option for localized prostate cancer (PCa), which is applied for over 15 years. There are conflicting recommendations for HIFU among urological societies, which can be explained by the lack of prospective controlled studies, reports on preselected patient populations and limited follow-up providing little information on overall and cancer-specific survival. We report on a large, unselected consecutive patient series of patients who have undergone primary HIFU for clinically localized PCa with the longest follow-up in current literature. Our results improve the understanding of the oncological efficacy, morbidity and side effects of primary HIFU. To assess the safety, functional and oncological long-term outcomes of high-intensity focused ultrasound (HIFU) as a primary treatment option for localized prostate cancer (PCa). We conducted a retrospective single-centre study on 538 consecutive patients who underwent primary HIFU for clinically localized PCa between November 1997 and September 2009. Factors assessed were: biochemical disease-free survival (BDFS) according to Phoenix criteria (prostate-specific antigen nadir + 2 ng/mL); metastatic-free, overall and PCa-specific survival; salvage treatment; side effects; potency; and continence status. The mean (sd; range) follow-up was 8.1 (2.9; 2.1-14.0) years. The actuarial BDFS rates at 5 and 10 years were 81 and 61%, respectively. The 5-year BDFS rates for low-, intermediate- and high-risk patients were 88, 83 and 48%, while the 10-year BDFS rates were 71, 63 and 32%, respectively. Metastatic disease was reported in 0.4, 5.7 and 15.4% of low-, intermediate- and high-risk patients, respectively. The salvage treatment rate was 18%. Seventy-five (13.9%) patients died. PCa-specific death was registered in 18 (3.3%) patients (0, 3.8 and 11% in the low-, intermediate- and high-risk groups, respectively). Side effects included bladder outlet obstruction (28.3%), Grade I, II and III stress urinary incontinence (13.8, 2.4 and 0.7%, respectively) and recto-urethral fistula (0.7%). Preserved potency was 25.4% (in previously potent patients). The study demonstrates the efficacy and safety of HIFU for localized PCa. HIFU is a therapeutic option for patients of advanced age, in the low- or intermediate-risk groups, and with a life expectancy of ∼10 years.


Molecular genetic alterations in endometrioid carcinomas of the ovary: similar frequency of beta-catenin abnormalities but lower rate of microsatellite instability and PTEN alterations than in uterine endometrioid carcinomas.
Endometrioid carcinomas of the ovary closely resemble their uterine counterparts. It has been suggested that the former tumors have the same molecular alterations (microsatellite instability [MSI], PTEN, and beta-catenin) described in endometrioid carcinomas of the uterus. We analyzed 55 ovarian carcinomas, including 22 endometrioid, 18 clear cell, and 15 mixed types. MSI was detected in 5 of 39 cases (13%). MLH1 promoter hypermethylation was identified in 2 of the 5 MSI-positive tumors. PTEN was mutated in 5 of 54 cases (9%); of these, 3 had MSI and exhibited frameshift mutations in short-coding mononucleotide repeats. Beta-catenin nuclear expression was detected in 11 of 54 cases (20%) by immunostaining; of these, 7 exhibited CTNNB1 gene mutations. These alterations were found more frequently in endometrioid carcinomas than in tumors of the other 2 groups. Among the former tumors, MSI was detected in 3 of 17 cases (17.5%); PTEN mutations, in 3 of 21 (14%); and beta-catenin, in 8 of 21 (38%). The molecular alterations were found more often in tumors associated with endometriosis than in tumors without endometriosis. Six endometrioid tumors demonstrating matrix metalloproteinase-7 (MMP-7) immunoreactivity with nuclear accumulation of beta-catenin had good outcomes, in contrast to poor outcomes in 7 of 9 predominantly nonendometrioid tumors demonstrating expression of MMP-7 only. We found a similar frequency of beta-catenin abnormalities but lower rates of MSI and PTEN alterations than in uterine endometrioid carcinomas. Alterations in beta-catenin and PTEN genes, as well as MSI, are frequent in low-stage ovarian carcinomas of endometrioid type that have a favorable prognosis.


Ventilatory chemoreflexes at rest following a brief period of heavy exercise in man.
1. Ventilatory chemoreflex responses have been studied at rest during the recovery from a brief period of heavy exercise. 2. Six young, healthy male subjects each undertook four experimental studies. In each study measurements were made at rest during recovery from an exhaustive 1-2 min sprint on a bicycle ergometer with a workload of 400 W. Three levels of end-tidal O2 pressure (Po2) were employed. Continuous ventilatory measurements were made during euoxia (end-tidal Po2, 100 Torr), hypoxia (end-tidal Po2, 50 Torr) and hyperoxia (end-tidal Po2, 300 Torr). Arterialized venous blood samples were drawn during each of the measurement periods for the estimation of arterial pH. In two of the studies, end-tidal CO2 pressure (Pco2) was maintained throughout at 1-2 Torr above the eucapnic level that existed prior to exercise (isocapnic post-exercise protocol, IPE). In the other two studies, end-tidal Pco2 was allowed to vary (poikilocapnic post-exercise protocol, PPE). Data from a previously published study on the same subjects involving an infusion of hydrochloric acid were used to provide control data with a varying level of metabolic acidosis, but with no prior exercise. 3. Ventilation-pH slopes in the IPE protocol were no different from control. Ventilation-pH slopes in the PPE protocol were significantly lower than in the IPE and control protocols (P < 0.05, ANOVA). This difference may be due to the progressive change in end-tidal Pco2 in the PPE protocol compared with the constant end-tidal Pco2 in the IPE and control protocols. 4. An arterial pH value of 7.35 was attained 30.4 +/- 2.7 min (mean +/- S.E.M.) after the end of exercise in the IPE protocol and 17.1 +/- 1.4 min after the end of exercise in the PPE protocol. 5. Hypoxic sensitivities at an arterial pH of 7.35 were not significantly different between the IPE, PPE and control protocols (ANOVA). 6. Euoxic ventilation at an arterial pH 7.35 was significantly greater than control for the IPE protocol (P < 0.001, Student's paired t test) and no different from control for the PPE protocol. 7. The results suggest that 30 min after heavy exercise, ventilation remains stimulated by processes other than the post-exercise metabolic acidosis, and that changes in peripheral chemoreflex sensitivity to hypoxia and acid are not implicated in this.


Angiotensin II causes cellular proliferation in infantile haemangioma via angiotensin II receptor 2 activation.
To investigate the effect of the angiotensin peptides and their agonists and antagonists on cellular proliferation in proliferating infantile haemangioma (IH) in vitro explants. Proliferating IH samples from six patients were cultured in vitro in the presence of angiotensin I (ATI) alone, or AT1 and the ACE inhibitor, ramipril, or ATII alone, or ATII with the ATII receptor 1 (ATIIR1) blocker, losartan, or ATII with the ATIIR2 blocker, PD123319, or the ATIIR2 agonist, CGP42112. After 6 days in culture, the IH tissue pieces were harvested, formalin-fixed and paraffin-embedded. The effect of each treatment type on cellular proliferation was evaluated by immunohistochemical staining of these tissue pieces using the proliferation marker, Ki67. There was a significant increase in cellular proliferation in the ATI and ATII treated IH tissues compared with control samples. Their effect on cellular proliferation was reduced by adding ramipril and PD123319, respectively. CGP42112, but not losartan, significantly increased cellular proliferation. Our findings suggest a key regulatory role of ATI and ATII in promoting cellular proliferation in IH, and establish a role for ACE and ATIIR2 in the proliferation of this tumour.


Motor imagery for severely motor-impaired patients: evidence for brain-computer interfacing as superior control solution.
Brain-Computer Interfaces (BCIs) strive to decode brain signals into control commands for severely handicapped people with no means of muscular control. These potential users of noninvasive BCIs display a large range of physical and mental conditions. Prior studies have shown the general applicability of BCI with patients, with the conflict of either using many training sessions or studying only moderately restricted patients. We present a BCI system designed to establish external control for severely motor-impaired patients within a very short time. Within only six experimental sessions, three out of four patients were able to gain significant control over the BCI, which was based on motor imagery or attempted execution. For the most affected patient, we found evidence that the BCI could outperform the best assistive technology (AT) of the patient in terms of control accuracy, reaction time and information transfer rate. We credit this success to the applied user-centered design approach and to a highly flexible technical setup. State-of-the art machine learning methods allowed the exploitation and combination of multiple relevant features contained in the EEG, which rapidly enabled the patients to gain substantial BCI control. Thus, we could show the feasibility of a flexible and tailorable BCI application in severely disabled users. This can be considered a significant success for two reasons: Firstly, the results were obtained within a short period of time, matching the tight clinical requirements. Secondly, the participating patients showed, compared to most other studies, very severe communication deficits. They were dependent on everyday use of AT and two patients were in a locked-in state. For the most affected patient a reliable communication was rarely possible with existing AT.


Epidemiology of bee stings in Campina Grande, Paraíba state, Northeastern Brazil.
The present study aims to investigate the clinical-epidemiological characteristics of bee sting cases recorded between 2007 and 2012 in the city of Campina Grande, Paraíba state, Brazil. Data were collected from the database of the Injury Notification Information System of the Brazilian Ministry of Health. A total of 459 bee sting cases were retrospectively analyzed. The average annual incidence was 19 cases per 100,000 inhabitants. Cases were distributed in all months of the year, with higher prevalence in September and February. Most victims were men aged between 20 and 29 years. The highest incidence of cases was recorded in urban areas. Victims were stung mainly on the head and torso and received medical assistance predominantly 1 to 3 hours after being stung. The most frequent clinical manifestations were pain, edema and itching. Most cases were classified as mild, and three deaths were reported. The high incidence of envenomations provoked by bees in Campina Grande suggests that it may be an important risk area for accidents. Since several medical records lacked information, clinical-epidemiological profile of bee sting cases in the studied region could not be accurately determined. The current study provides relevant data for the development of strategies to promote control and prevention of bee stings in this area. Further training for health professionals seems to be necessary to improve their skills in recording clinical-epidemiological information as well as in treating bee sting victims.


LILRB4-targeting Antibody-Drug Conjugates for the Treatment of Acute Myeloid Leukemia.
Acute myeloid leukemia (AML) is the most common and aggressive blood cancer in adults. In particular, significant unmet medical needs exist for effective treatment strategies for acute myelomonocytic leukemia (M4) and acute monocytic leukemia (M5) AML subtypes. Antibody-drug conjugates (ADC) are a promising drug class for AML therapy, as demonstrated by the FDA-approved anti-CD33 ADC, gemtuzumab ozogamicin (Mylotarg). However, CD33 is expressed in normal hematopoietic stem cells, highlighting the critical need to identify AML-specific targets to minimize the risk of potential adverse effects. We have demonstrated that the leukocyte immunoglobulin-like receptor subfamily B4 (LILRB4) is expressed at significantly higher levels on monocytic M4 and M5 AML cells than on normal counterparts. Here, we test whether LILRB4 is a promising ADC target to kill monocytic AML cells while sparing healthy counterparts. To this end, we generated ADCs from a humanized anti-LILRB4 mAb and the antimitotic payload, monomethyl auristatin F. The conjugates constructed were characterized and evaluated for LILRB4-specific cell killing potency, toxicity to progenitor cells, pharmacokinetics, and therapeutic efficacy. Our ADC linker technology platform efficiently generated homogeneous anti-LILRB4 ADCs with defined drug-to-antibody ratios. The homogeneous anti-LILRB4 ADCs demonstrated the capacity for LILRB4-mediated internalization, suitable physicochemical properties, and high cell killing potency against LILRB4-positive AML cells. Importantly, our data indicate that these ADCs spare normal progenitor cells. One of our homogeneous conjugates exerted a remarkable therapeutic effect and no significant toxicity in a xenograft mouse model of disseminated human AML. Our findings highlight the clinical potential of anti-LILRB4 ADCs in monocytic AML therapy.


Comparison of conservation metrics in a case study of lemurs.
Conservation planning is important to protect species from going extinct now that natural habitats are decreasing owing to human activity and climate change. However, there is considerable controversy in choosing appropriate metrics to weigh the value of species and geographic regions. For example, the added value of phylogenetic conservation-selection criteria remains disputed because high correlations between them and the nonphylogenetic criteria of species richness have been reported. We evaluated the commonly used conservation metrics species richness, endemism, phylogenetic diversity (PD), and phylogenetic endemism (PE) in a case study on lemurs of Madagascar. This enabled us to identify the conservation target of each metric and consider how they may be used in future conservation planning. We also devised a novel metric that uses a phylogeny scaled according to the rate of phenotypic evolution as a proxy for a species' ability to adapt to change. High rates of evolution may indicate generalization or specialization. Both specialization and low rates of evolution may result in an inability to adapt to changing environments. We examined conservation priorities by using the inverse of the rate of body mass evolution to account for species with low rates of evolution. In line with previous work, we found high correlations among species richness and PD (r = 0.96), and endemism and PE (r = 0.82) in Malagasy lemurs. Phylogenetic endemism in combination with rates of evolution and their inverse prioritized grid cells containing highly endemic and specialized lemurs at risk of extinction, such as Avahi occidentalis and Lepilemur edwardsi, 2 endangered lemurs with high rates of phenotypic evolution and low-quality diets, and Hapalemur aureus, a critically endangered species with a low rate of body mass evolution and a diet consisting of very high doses of cyanide.


Folic acid supplementation and cell kinetics of rectal mucosa in patients with ulcerative colitis.
It has been suggested that colon cancer risk in ulcerative colitis (UC) is correlated to a reduced bioavailability of folate. We studied the effects of folate supplementation on the pattern of rectal cell proliferation in patients affected by long-standing UC. In the rectal mucosa of these patients, an expansion of proliferating cells to the crypt surface is found frequently. This abnormality is considered an intermediate biomarker in chemoprevention trials. Twenty-four patients (13 males; age, 26-70 years; UC duration, 7-34 years) with UC in remission for 1 month at least were assigned randomly to one of the following treatments: (a) folinic acid (15 mg/day) or (b) placebo. Cell proliferation was analyzed through immunohistochemistry on sections of rectal biopsies incubated for 1 hour in a culture medium containing bromodeoxyuridine. Fragments were taken at admission to the study and after 3 months of treatment. As compared to the baseline values, after 3 months of therapy in patients treated with folinic acid, a significant reduction of the frequency of occurrence of labeled cells in the upper 40% of the crypts (phi h value) was observed (0.1836 +/- 0.0278 versus 0.1023 +/- 0.0255; P < 0.01). On the contrary, no significant proliferative changes were observed in the placebo group. These results suggest that folate supplementation contributes to regulating rectal cell proliferation in patients with long-standing UC. These findings may be significant for chemoprevention of colon cancer in these patients.


pH dependence of myosin binding-induced activation of the thin filament in cardiac myocytes and skeletal fibers.
The pH dependence of myosin binding-induced thin filament activation was determined in permeabilized cardiac myocytes and slow- and fast-twitch single skeletal muscle fibers by experimental lowering of [MgATP] in the Ca(2+)-free solutions bathing the permeabilized preparations. As the pS (where S is [MgATP] and pS is -log[MgATP]) was increased from 3.0 to 8.0, isometric tension increased to a peak value in the pS range of 4.9-5.3. At pH 7.00, the transition from the relaxed to the activated rigor state was steep in cardiac myocytes [Hill value (nH) = 21.2 +/- 3.1 (SE)] and due to the apparent effect of strongly bound cross bridges to cooperatively activate the thin filament in the absence of added Ca2+. At pH 6.20, the steepness of the tension-pS relationship was markedly reduced (nH = 6.1 +/- 1.0) and the midpoint of the relationship (pS50) was shifted to higher pS values in cardiac myocytes. In comparison, reduced pH had no effect on the steepness or position of the tension-pS relationship in single slow- or fast-twitch skeletal muscle fibers. These findings suggest that myosin binding-induced activation of the thin filament is pH dependent in cardiac myocytes but not in skeletal muscle fibers under these experimental conditions in which Ca2+ is absent.


The role of neck dissection in the combined modality therapy setting.
No method reliably detects residual low-volume nodal disease in a patient with a complete response (CR) to treatment. Control of the N0-1 neck is to be expected after CR; no treatment is needed. Positron emission tomography (PET) may improve patient selection, but neck dissection should still be performed for patients with good performance status and residual masses. Neck dissection reduces the incidence of regional recurrence, although the impact on survival is small. Whether the risk of tumor recurrence or the morbidity of neck dissection should be of greater concern for patients with N2-3 disease in CR is a matter of individual judgment. Combined modality therapy will control most nodal metastases (even for patients with advanced nodal disease). Neck dissection in patients presenting with bulky nodal disease who achieve a CR after combined modality therapy will diminish the regional recurrence rate by about 15%. Nowadays, patients with N3 disease and CR often still have residual neck tumor. Node dissection is to be advised. Neck dissection should still be considered an important treatment modality for patients who undergo combined modality therapy with advanced nodal disease, even if they have achieved a CR.


The alkyltransferase-like ybaZ gene product enhances nucleotide excision repair of O(6)-alkylguanine adducts in E. coli.
O(6)-methylguanine adducts are potent pre-mutagenic lesions owing to their high capacity to direct mis-insertion of thymine when bypassed by replicative DNA polymerases. The strong mutagenic potential of these adducts is prevented by alkyltransferases such as Ada and Ogt in Escherichia coli that transfer the methyl group to one of their cysteine residues. Alkyl residues larger than methyl are generally weak substrates for reversion by alkyltransferases. In this paper we have investigated the genotoxic potential of the O(6)-alkylguanine adducts formed by ethylene and propylene oxide using single-adducted plasmid probes. Our work shows that the ybaZ gene product, a member of the alkyltransferase-like protein family, strongly enhances the repair by nucleotide excision repair of the larger O(6)-alkylguanine adducts that are otherwise poor substrates for alkyltransferases. The YbaZ protein is shown to interact with UvrA. This factor may thus enhance the efficiency of nucleotide excision repair in a way similar to the Transcription-Repair Coupling factor Mfd, by recruiting the UvrA(2).UvrB complex to the adduct site via its interaction with UvrA.


Genetic interplay between HLA-C and MIR148A in HIV control and Crohn disease.
Variation in the 3' untranslated region (3'UTR) of the HLA-C locus determines binding of the microRNA Hsa-miR-148a, resulting in lower cell surface expression of alleles that bind miR-148a relative to those alleles that escape its binding. The HLA-C 3'UTR variant was shown to associate with HIV control, but like the vast majority of disease associations in a region dense with causal candidates, a direct effect of HLA-C expression level on HIV control was not proven. We demonstrate that a MIR148A insertion/deletion polymorphism associates with its own expression levels, affecting the extent to which HLA-C is down-regulated, the level of HIV control, and the risk of Crohn disease only among those carrying an intact miR-148a binding site in the HLA-C 3'UTR. These data illustrate a direct effect of HLA-C expression level on HIV control that cannot be attributed to other HLA loci in linkage disequilibrium with HLA-C and highlight the rich complexity of genetic interactions in human disease.


Cell surface display system for Lactococcus lactis: a novel development for oral vaccine.
The food-grade Lactococcus lactis is a potential vector to be used as a live vehicle for the delivery of heterologous proteins for vaccine and pharmaceutical purposes. We constructed a plasmid vector pSVac that harbors a 255-bp single-repeat sequence of the cell wall-binding protein region of the AcmA protein. The recombinant plasmid was transformed into Escherichia coli and expression of the gene fragment was driven by the T7 promoter of the plasmid. SDS-PAGE showed the presence of the putative AcmA' fragment and this was confirmed by Western blot analysis. The protein was isolated and purified using a His-tag affinity column. When mixed with a culture of L. lactis MG1363, ELISA and immunofluorescence assays showed that the cell wall-binding fragment was anchored onto the outer surface of the bacteria. This indicated that the AcmA' repeat unit retained the active site for binding onto the cell wall surface of the L. lactis cells. Stability assays showed that the fusion proteins (AcmA/A1, AcmA/A3) were stably docked onto the surface for at least 5 days. The AcmA' fragment was also shown to be able to strongly bind onto the cell surface of naturally occurring lactococcal strains and Lactobacillus and, with less strength, the cell surface of Bacillus sphericus. The new system designed for cell surface display of recombinant proteins on L. lactis was evaluated for the expression and display of A1 and A3 regions of the VP1 protein of enterovirus 71 (EV71). The A1 and A3 regions of the VP1 protein of EV71 were cloned upstream to the cell wall-binding domains of AcmA protein and successfully expressed as AcmA/A1 and AcmA/A3. Whole-cell ELISA showed the successful display of VP1 protein epitopes of EV71 on the surface of L. lactis. The success of the anchoring system developed in this study for docking the A1 and A3 epitopes of VP1 onto the surface of L. lactis cells opens up the possibilities of peptide and protein display for not only Lactococcus but also for other gram-positive bacteria. This novel way of displaying epitopes on the cell surface of L. lactis and other related organisms should be very useful in the delivery of vaccines and other useful proteins.


Pecans lower low-density lipoprotein cholesterol in people with normal lipid levels.
To compare serum lipid profiles and dietary intakes of people with normal lipid levels who consumed pecans and those who did not consume nuts. Eight-week, randomized, controlled study of pecan treatment group vs control group. Nineteen people with normal lipid levels completed the study; 10 had been randomly assigned to the pecan treatment group (7 women, 3 men, mean age = 45 +/- 10 years) and 9 to the control group (8 women, 1 man, mean age = 37 +/- 12 years). The pecan treatment group consumed 68 g pecans per day for 8 weeks plus self-selected diets. The pecans contributed 459 kcal and 44 g fat daily. The control group avoided nuts and consumed self-selected diets. Total serum cholesterol, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and total triglyceride levels were measured at the time of entrance to the study (baseline), week 4, and week 8. Computer analyses were done on five 3-day food records. Comparisons were made using analysis of variance or paired t test. LDL-C was lowered in the pecan treatment group from 2.61 +/- 0.49 mmol/L at baseline to 2.35 +/- 0.49 at week 4 (P < .05) and to 2.46 +/- 0.59 at week 8 (P < .05). At week 8, total cholesterol and HDL-C in the pecan treatment group were significantly lower (P < .05) than in the control group (total cholesterol: 4.22 +/- 0.83 vs 5.02 +/- 0.54 mmol/L; HDL-C: 1.37 +/- 0.23 vs 1.47 +/- 0.34 mmol/L). Dietary fat, monounsaturated fat, polyunsaturated fat, insoluble fiber, magnesium, and energy were significantly higher in the pecan treatment group than in the control group. Body mass indexes and body weights were unchanged in both groups. Pecans can be included in a healthful diet when energy intake and potential weight gain are addressed.


Comparison of Image Quality of Different Radionuclides Technetium-99m, Samarium-153, and Iodine-123.
The choice of the radionuclide has a key role in nuclear medicine which appearing the lowest scatter fraction. In addition, the presence of penetrated and scattered photons from collimator in single-photon emission computed tomography images degrades resolution and contrast. Thus, image quality depends on sensitivity and resolution of the collimator-detector system. The goal of this study was to compare the image quality that can be achieved by three radionuclides: technetium-99 m (Tc-99 m), iodine-123 (I-123), and samarium-153 (Sm-153). Tc-99 m and Sm-153 were imaged with low-energy high resolution (LEHR) collimator, while I-123 was imaged with medium-energy (ME) collimator. We modeled the Siemens Symbia Medical system using Monte Carlo simulation code SIMIND. The imaging characteristics of each radionuclide were investigated by simulated data: point spread function, sensitivity (Cps/MBq) and geometric, penetration and scattering distribution. Tc-99 m and Sm-153 give best and results with LEHR collimator for spatial resolution (full width at half maximum [FWHM] = 3.19 mm; full width at tenth maximum [FWTM] = 6.73 mm) and (FWHM = 3.22 mm; FWTM = 7.39 mm), respectively. Whereas, I-123 provided with ME collimator a lower resolution (FWHM = 4.89 mm; FWTM = 9.89 mm). The sensitivity recorded by Tc-99 m, Sm-153, and I-153 were (31.21 Cps/MBq), (10.16 Cps/MBq), and (51.22 Cps/MBq), respectively. Tc-99 m and Sm-153 give the best and generally similar imaging properties with LEHR. For I-123, the ME collimator helps lowering the influence of high-energy gamma rays.


Dephosphorylated synapsin I anchors synaptic vesicles to actin cytoskeleton: an analysis by videomicroscopy.
Synapsin I is a synaptic vesicle-associated protein which inhibits neurotransmitter release, an effect which is abolished upon its phosphorylation by Ca2+/calmodulin-dependent protein kinase II (CaM kinase II). Based on indirect evidence, it was suggested that this effect on neurotransmitter release may be achieved by the reversible anchoring of synaptic vesicles to the actin cytoskeleton of the nerve terminal. Using video-enhanced microscopy, we have now obtained experimental evidence in support of this model: the presence of dephosphorylated synapsin I is necessary for synaptic vesicles to bind actin; synapsin I is able to promote actin polymerization and bundling of actin filaments in the presence of synaptic vesicles; the ability to cross-link synaptic vesicles and actin is specific for synapsin I and is not shared by other basic proteins; the cross-linking between synaptic vesicles and actin is specific for the membrane of synaptic vesicles and does not reflect either a non-specific binding of membranes to the highly surface active synapsin I molecule or trapping of vesicles within the thick bundles of actin filaments; the formation of the ternary complex is virtually abolished when synapsin I is phosphorylated by CaM kinase II. The data indicate that synapsin I markedly affects synaptic vesicle traffic and cytoskeleton assembly in the nerve terminal and provide a molecular basis for the ability of synapsin I to regulate the availability of synaptic vesicles for exocytosis and thereby the efficiency of neurotransmitter release.


Controllable dynamics of oxygen vacancies through extrinsic doping for superior catalytic activities.
Due to its strong redox ability, high stability, cost effectiveness and non-toxicity, cerium oxide (CeO2) has been extensively researched as an active photocatalyst material. The underlying photocatalytic reactions are mostly associated with the transportation of oxygen ions through vacancies, but the actual transport phenomenon had not been clearly understood. In this work, gadolinium (Gd) is sequentially doped into CeO2 to investigate how extrinsic doping can modulate oxygen vacancies in CeO2 and influence photocatalytic activities. From our investigations, it was found that the Gd doping may induce structural symmetry breaking leading to a pure CeO2 fluorite structure that transforms mobile oxygen vacancies into clustered or immobile vacancies. When the vacancies were set as "mobile" (for Gd doping levels ≤15 at%), maximum photocatalytic activities were obtained. In contrast, suppressed photocatalytic efficiencies were noted for higher Gd doping levels (20 at% or more). The results reported in this research may provide an extra degree of freedom in the form of extrinsic doping to configure the oxygen vacancy defects and their mobility to achieve better catalytic efficiencies.


Restriction fragment length polymorphism analysis of Listeria monocytogenes and its application to epidemiological investigations.
The restriction fragment length polymorphisms (RFLPs) of 64 random and potentially related strains of Listeria monocytogenes were analysed and compared using a probe comprised of two L. monocytogenes chromosome fragments cloned into a lambda vector. Twelve RFLP types were defined using 14 isolates of clinical origin, 42 food isolates and eight food associated environmental strains. Of the RFLP types, some were common to a particular serovar and source, whereas others were widespread amongst all serovars and sources. One of the two most common RFLP patterns was associated with serovar 1/2 isolates from food or the environment, whereas another dominant pattern was associated most commonly with serovar four isolates from all sources. The potential relationships between epidemiologically related strains were examined, with the analysis of types from a suspected listeriosis outbreak, from clinical maternal-foetal cases, and from an ice-cream factory environmental study. Serotyping alone was not a sufficient marker for the comparison of these strains whereas further discrimination of strains was possible with RFLP analysis.


Deciphering the role of the type II glyoxalase isoenzyme YcbL (GlxII-2) in Escherichia coli.
In Escherichia coli, detoxification of methylglyoxal (MG) requires glyoxalases I and II. Glyoxalase I (gloA/GlxI) isomerizes the hemithioacetal, formed spontaneously from MG and glutathione (GSH) to S-lactoylglutathione (SLG), which is hydrolyzed by glyoxalase II (gloB/GlxII) to lactate and GSH. YcbL from Salmonella enterica serovar Typhimurium is an unusual type II glyoxalase whose role in MG detoxification has remained enigmatic. Here we show that YcbL (gloC/GlxII-2) acts as an accessory type II glyoxylase in E. coli. The two isoenzymes have additive effects and ensure maximal MG degradation.


Effect of salinity on mercury methylating benthic microbes and their activities in Great Salt Lake, Utah.
Surface water and biota from Great Salt Lake (GSL) contain some of the highest documented concentrations of total mercury (THg) and methylmercury (MeHg) in the United States. In order to identify potential biological sources of MeHg and controls on its production in this ecosystem, THg and MeHg concentrations, rates of Hg(II)-methylation and MeHg degradation, and abundances and compositions of archaeal and bacterial 16 rRNA gene transcripts were determined in sediment along a salinity gradient in GSL. Rates of Hg(II)-methylation were inversely correlated with salinity and were at or below the limits of detection in sediment sampled from areas with hypersaline surface water. The highest rates of Hg(II)-methylation were measured in sediment with low porewater salinity, suggesting that benthic microbial communities inhabiting less saline environments are supplying the majority of MeHg in the GSL ecosystem. The abundance of 16S rRNA gene transcripts affiliated with the sulfate reducer Desulfobacterium sp. was positively correlated with MeHg concentrations and Hg(II)-methylation rates in sediment, indicating a potential role for this taxon in Hg(II)-methylation in low salinity areas of GSL. Reactive inorganic Hg(II) (a proxy used for Hg(II) available for methylation) and MeHg concentrations were inversely correlated with salinity. Thus, constraints imposed by salinity on Hg(II)-methylating populations and the availability of Hg(II) for methylation are inferred to result in higher MeHg production potentials in lower salinity environments. Benthic microbial MeHg degradation was also most active in lower salinity environments. Collectively, these results suggest an important role for sediment anoxia and microbial sulfate reducers in the production of MeHg in low salinity GSL sub-habitats and may indicate a role for salinity in constraining Hg(II)-methylation and MeHg degradation activities by influencing the availability of Hg(II) for methylation.


Cyclopia and defective axial patterning in mice lacking Sonic hedgehog gene function.
Targeted gene disruption in the mouse shows that the Sonic hedgehog (Shh) gene plays a critical role in patterning of vertebrate embryonic tissues, including the brain and spinal cord, the axial skeleton and the limbs. Early defects are observed in the establishment or maintenance of midline structures, such as the notochord and the floorplate, and later defects include absence of distal limb structures, cyclopia, absence of ventral cell types within the neural tube, and absence of the spinal column and most of the ribs. Defects in all tissues extend beyond the normal sites of Shh transcription, confirming the proposed role of Shh proteins as an extracellular signal required for the tissue-organizing properties of several vertebrate patterning centres.


A new mouse model for the study of human breast cancer metastasis.
Breast cancer is the most common cancer in women, and this prevalence has a major impact on health worldwide. Localized breast cancer has an excellent prognosis, with a 5-year relative survival rate of 85%. However, the survival rate drops to only 23% for women with distant metastases. To date, the study of breast cancer metastasis has been hampered by a lack of reliable metastatic models. Here we describe a novel in vivo model using human breast cancer xenografts in NOD scid gamma (NSG) mice; in this model human breast cancer cells reliably metastasize to distant organs from primary tumors grown within the mammary fat pad. This model enables the study of the entire metastatic process from the proper anatomical site, providing an important new approach to examine the mechanisms underlying breast cancer metastasis. We used this model to identify gene expression changes that occur at metastatic sites relative to the primary mammary fat pad tumor. By comparing multiple metastatic sites and independent cell lines, we have identified several gene expression changes that may be important for tumor growth at distant sites.


Repair of a coracoid luxation and a tibiotarsal fracture in a bald eagle (Haliaeetus leucocephalus).
A 4.5-kg, adult bald eagle (Haliaeetus leucocephalus) was presented after being found unable to fly. Crepitus was palpated in the left shoulder; however, the wing position was normal. Radiographs revealed caudoventral luxation of the left coracoid, affecting its distal articulation with the clavicle and its proximal articulation with the sternum. The luxation affecting the coracoid-sternum articulation was repaired by using a 4-hole, 1.5-mm T-plate and a 6-hole, 2.0-mm dynamic compression plate (DCP) placed side by side. The luxation affecting the coracoid and the clavicle was repaired by 2 cerclage wires in a simple interrupted pattern. Before a scheduled release, the bird sustained a closed, complete mid diaphyseal transverse fracture of the right tibiotarsus, which most likely occurred during recapture from the flight cage. The fracture was surgically repaired with 2 circumferential cerclage wires, an intramedullary Kirschner wire and one 10-hole, 2.7-mm DCP. The bird was successfully released, 105 days after the first surgery, near the location where it was found.


Role of down-regulated neutral ceramidase during all-trans retinoic acid-induced neuronal differentiation in SH-SY5Y neuroblastoma cells.
Neutral ceramidase (NCDase) is considered to be a critical enzyme for controlling the turnover of ceramide, an important bioactive lipid, which determines cell's fate. All-trans retinoic acid (ATRA) has been reported to induce neuronal differentiation and cell-cycle arrest [Lopez-Carballo, Moreno, Masia, Perez, and Barettino (Activation of the phosphatidylinositol 3-kinase/Akt signalling pathway by retinoic acid is required for neural differentiation of SH-SY5Y human neuroblastoma cells. J Biol Chem 2002:277:25297-304.)]. In this study, we observed that ATRA-induced cellular ceramide accumulation, cell-growth arrest and differentiation accompanied with down-regulation of NCDase in SH-SY5Y cells, without a decrease in sphingosine or sphingosine 1-phosphate. We examined whether the down-regulation of NCDase was involved in the increase in ceramide and cell differentiation. ATRA was found to down-regulate mRNA, protein and the enzyme activity of NCDase. Interestingly, GATA-2 was also decreased with ATRA treatment, and experiments using its expression vector and siRNA and chromatin immunoprecipitation assay demonstrated GATA-2 acted as transcription-factor of NCDase gene expression. By establishing stable transfectants with decreased NCDase expression and activity, we clarified the significance of NCDase down-regulation for ATRA-induced neuronal differentiation. Those sub-clones showed both increased cellular ceramide and reduced cell growth as well as neuronal differentiation phenotypes. These results demonstrate that down-regulation of NCDase through ATRA-induced GATA-2 decrease plays an important role in induction of ceramide accumulation and neuronal differentiation in SH-SY5Y cells.


High-performing dry powder inhalers of paclitaxel DPPC/DPPG lung surfactant-mimic multifunctional particles in lung cancer: physicochemical characterization, in vitro aerosol dispersion, and cellular studies.
Inhalable lung surfactant-based carriers composed of synthetic phospholipids, dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylglycerol (DPPG), along with paclitaxel (PTX), were designed and optimized as respirable dry powders using organic solution co-spray-drying particle engineering design. These materials can be used to deliver and treat a wide variety of pulmonary diseases with this current work focusing on lung cancer. In particular, this is the first time dry powder lung surfactant-based particles have been developed and characterized for this purpose. Comprehensive physicochemical characterization was carried out to analyze the particle morphology, surface structure, solid-state transitions, amorphous character, residual water content, and phospholipid bilayer structure. The particle chemical composition was confirmed using attenuated total reflectance-Fourier-transform infrared (ATR-FTIR) spectroscopy. PTX loading was high, as quantified using UV-VIS spectroscopy, and sustained PTX release was measured over weeks. In vitro cellular characterization on lung cancer cells demonstrated the enhanced chemotherapeutic cytotoxic activity of paclitaxel from co-spray-dried DPPC/DPPG (co-SD DPPC/DPPG) lung surfactant-based carrier particles and the cytotoxicity of the particles via pulmonary cell viability analysis, fluorescent microscopy imaging, and transepithelial electrical resistance (TEER) testing at air-interface conditions. In vitro aerosol performance using a Next Generation Impactor™ (NGI™) showed measurable powder deposition on all stages of the NGI and was relatively high on the lower stages (nanometer aerodynamic size). Aerosol dispersion analysis of these high-performing DPIs showed mass median diameters (MMADs) that ranged from 1.9 to 2.3 μm with excellent aerosol dispersion performance as exemplified by high values of emitted dose, fine particle fractions, and respirable fractions.


Prevalence and severity of atherosclerosis in extra cranial carotid arteries in Nigeria: an autopsy study.
There has been a paucity of autopsy studies on atherosclerotic lesions in Nigerians, the last one conducted at our centre being more than four decades ago. There has also been considerable epidemiological transition. The objective of the study was to determine the frequency, severity, pattern and distribution of atherosclerotic lesions in extra cranial carotid arteries (ECCA) in Nigerians at autopsy. ECCA of 30 consecutive Nigerian patients undergoing autopsy at a University teaching hospital were examined using the American Heart Association (AHA) histological grading and classification of atherosclerosis. Atherosclerotic lesions of ECCA were present in 73.3% of the subjects with the right and the left carotid bifurcations (28.3%) being the most frequently affected sites. Using the AHA classification of atherosclerosis, a total of 176(73.3%) lesions were found in the 240 histological sections of blood vessels examined. Of these, 22.5% were types I, 22.5% were types II, 15.4% were type V, and 7.5% were type III. The VII to type IX lesions were rare. When these atherosclerotic lesions were grouped into mild, moderate and severe, 52.5% were mild lesions (types I-III); 18.3% were moderate lesions (types IV and V); and 2.5% were severe lesions (types VI to IX). The severe lesions were most frequently observed in the left carotid bifurcation (50%) and they first appeared in the age group 45-49 years. Age, hypertension and diabetes mellitus were strong risk factors for atherosclerosis. Compared with four decades ago there has been an apparent increase in severity and extent of ECCA atherosclerosis especially after the age of 45 years in autopsies from our centre. This change in the amount of atherosclerosis over time is possibly due to the epidemiologic transition. This may worsen the rise in stoke incidence within this community and as such, great effort should be made to follow-up and manage CVD risk factors within the community.


Development of novel predictive miRNA/target gene pathways for colorectal cancer distance metastasis to the liver using a bioinformatic approach.
Liver metastases are the major cause of colorectal cancer (CRC)-related deaths. However, there is no reliable clinical predictor for CRC progression to liver metastasis. In this study, we investigated possible predictors (miRNAs and biomarkers) for clinical application. The Gene Expression Omnibus (GEO) datasets GSE49355, GSE41258 and GSE81558 for genes and GSE54088 and GSE56350 for miRNAs were used to identify common differentially expressed genes (DEGs) and miRNAs between primary CRC tissues and liver metastases. The identified miRNAs and their targets from the DEGs were verified in datasets comprising gene, miRNA and miRNA exosome profiles of CRC patients with no distant metastases (M0) and distant metastases (M1); the interaction networks and pathways were also mapped. There were 49 upregulated and 13 downregulated DEGs and 16 downregulated and 14 upregulated miRNAs; between the DEGs and miRNA targets, there were five upregulated and four downregulated genes. MiR-20a was strongly correlated with the status of liver metastasis. MiR-20a, miR499a, and miR-576-5p were highly correlated with the metastatic outcomes. MiR-20a was significantly highly expressed in the M1 group. In an analysis of the miRNA target genes, we found that CDH2, KNG1, and MMP2 were correlated with CRC metastasis. We demonstrated a new possible pathway for CRC metastasis: miR-576-5p/F9, miR20a/MMP2, CTSK, MMP3, and miR449a/P2RY14. The regulation of IGF transport and uptake by IGFBPs, extracellular matrix organization, signal transduction and the immune system were the enriched pathways. This model can predict CRC to liver metastases and the pathways involved, which can be clinically applicable.


Mean level signal crossing rate for an arbitrary stochastic process.
The issue of the mean signal level crossing rate for various probability density functions with primary relevance for optics is discussed based on a new analytical method. This method relies on a unique transformation that transforms the probability distribution under investigation into a normal probability distribution, for which the distribution of mean level crossings is known. In general, the analytical results for the mean level crossing rate are supported and confirmed by numerical simulations. In particular, we illustrate the present method by presenting analytic expressions for the mean level crossing rate for various probability distributions, including ones that previously were unavailable, such as the uniform, the so-called gamma-gamma, and the Rice-Nakagami distribution. However, in a limited number of cases the present results differ somewhat from the result reported in the literature. At present, this discrepancy remains unexplained and is laid open for future discussion.


Selection and adaptation in the human genome.
An enduring goal of evolutionary biology is to understand how natural selection has shaped patterns of polymorphism and divergence within and between species and to map the genetic basis of adaptations. The rapid maturation of next-generation sequencing technology has generated a deluge of genomics data from nonhuman primates, extinct hominins, and diverse human populations. These emerging genome data sets have simultaneously broadened our understanding of human evolution and sharply defined existing gaps in knowledge about the mechanistic basis of evolutionary change. In this review, we summarize recent insights into how natural selection has influenced the human genome across different timescales. Although the path to a more comprehensive understanding of selection and adaptation in humans remains arduous, some general insights are beginning to emerge, such as the importance of adaptive regulatory evolution, the absence of pervasive classic selective sweeps, and the potential roles that selection from standing variation and polygenic adaptation have likely played in recent human evolutionary history.


Pimecrolimus, a topical calcineurin inhibitor used in the treatment of atopic eczema.
Pimecrolimus , a calcineurin inhibitor, is a non-steroidal treatment option in patients aged ≥ 2 years with mild-to-moderate atopic eczema (AE). It was approved as a viable therapeutic option by the FDA in 2001 and in the European Union a year later in 2002. Calcineurin inhibitors inhibit the synthesis of inflammatory cytokines released from T cells and mast cells. In contrast to corticosteroids, calcineurin inhibitors act specifically on proinflammatory cells. Pimecrolimus shows comparative efficacy to mild topical corticosteroids and a special antipruritic effect. Furthermore, examinations of the systemic absorption of pimecrolimus implicated no systemic immunosuppression. In 2006, the FDA set a black box warning in the packaging materials of pimecrolimus alluding to the risk of skin malignancy or lymphomas due to theoretical consideration. The authors provide a review of pimecrolimus as a treatment for AE. Specifically, the authors present the pharmacokinetic and pharmacodynamic information on pimecrolimus and also review its efficacy. The authors also discuss pimecrolimus' safety and tolerability profile. Pimecrolimus represents a valuable part of active and proactive therapy in AE. That being said, the long-term safety of topical calcineurin inhibitors remains to be investigated. Given the results from experimental photocarcinogenicity studies, effective sun protection should be employed during the therapy, although an increased risk for skin malignancies and lymphomas was not found in recent studies. Pimecrolimus should be considered as an alternative therapeutic approach in AE treatment management going along with a corticoid-sparing effect.


Ignoring it doesn't make it stop: adolescents, appearance, and bullying.
To investigate levels of appearance-related concern in a normative population of adolescents. Action research methodology was used to help adolescents develop a questionnaire that elicited peer views on appearance (Part 1) and to further develop the Changing Faces psychosocial strategies for dealing with appearance-related bullying (Part 2). Adolescents aged 11-19.5 years were involved in the development of a questionnaire on the extent and nature of appearance-related concerns: 36 in development of antibullying strategies and 210 in an intervention. Part 3 of the project was an intervention that included information on the importance of appearance in human interactions as well as learning and practicing eight nonconfrontational coping strategies. The questionnaires developed by the adolescents, in addition to Rosenberg's Self-Esteem Questionnaire for Adolescents, were used preintervention, postintervention, and at 6 months postintervention. Of 11-13 year olds questioned, 75% cited teasing or bullying about their appearance as causing considerable distress. Concerns were compounded by a perceived lack of effective coping strategies. At 6 months postintervention, perceived levels of bullying had decreased by almost two-thirds, and there were significant improvements in global self-esteem, confidence to tackle teasing and bullying, and confidence with disfigurement issues. A nonintervention comparison group showed no such improvements. Offering young people social skills to cope with teasing or bullying about appearance may substantially reduce general bullying as well as that specifically targeted at disfigured individuals. Involving young people in the design of an intervention to be used with them may be crucial to the intervention's eventual success, and strategies learned to cope with appearance-related bullying are easily adaptable for use in other types of confrontation.


[Complementary obstetrical delivery methods].
Complementary methods in obstetrics are related to humanization of delivery and woman's decision on the mode of delivery in normal labor. The methods include various birthing positions, aids such as delivery chair, birthing wheel, etc., and water delivery. Results of recent studies comparing these methods with classic delivery in supine position are presented. The advantages of these alternative methods of delivery include shorter duration of delivery, reduced need of labor induction, lower use of analgesics, and women's acceptance of these methods of delivery with the same level of maternal and neonatal safety as in classic delivery. All these advantages apply to water delivery providing the prerequisities, contraindications and measures of surveillance are strictly met.


Randomized, cross-over evaluation of mobile phone vs paper diary in subjects with mild to moderate persistent asthma.
Diaries are frequently used to evaluate therapy. Forgetfulness, however, can lead to missed entries. With paper diaries, these missing entries can be backfilled, compromising the reasons for using a diary. Electronic diaries are a potential means of mitigating this limitation. The pilot study was conducted to evaluate use of a mobile phone diary. Twelve subjects with mild persistent asthma were randomly assigned to mobile or paper diary for 2 weeks and then crossed over to use the other diary type for next 2 weeks. Of the 12 subjects, 7 preferred the mobile diary. However, the mean prevalence of missing data was greater when using the mobile (18% ± 9%) compared to paper diary (9% ± 4%; P = 0.05). In conclusion, the mobile diary was preferred by slightly more subjects. The greater prevalence of missing data when using this diary most likely results from the inability to backfill missing entries. Clintrials.gov NCT00367263 (http://clinicaltrials.gov/ct2/show/NCT00367263).


Candle flame soot sizing by planar time-resolved laser-induced incandescence.
Soot emissions from flaming combustion are relevant as a significant source of atmospheric pollution and as a source of nanomaterials. Candles are interesting targets for soot characterization studies since they burn complex fuels with a large number of carbon atoms, and yield stable and repeatable flames. We characterized the soot particle size distributions in a candle flame using the planar two-color time-resolved laser induced incandescence (2D-2C TiRe-LII) technique, which has been successfully applied to different combustion applications, but never before on a candle flame. Soot particles are heated with a planar laser sheet to temperatures above the normal flame temperatures. The incandescent soot particles emit thermal radiation, which decays over time when the particles cool down to the flame temperature. By analyzing the temporal decay of the incandescence signal, soot particle size distributions within the flame are obtained. Our results are consistent with previous works, and show that the outer edges of the flame are characterized by larger particles ([Formula: see text]), whereas smaller particles ([Formula: see text]) are found in the central regions. We also show that our effective temperature estimates have a maximum error of 100 K at early times, which decreases as the particles cool.


Laboratory evaluation of the toxicity of systemic insecticides for Control of Anoplophora glabripennis and Plectrodera scalator (Coleoptera: Cerambycidae).
Anoplophora glabripennis (Motschulsky) (Coleoptera: Cerambycidae) is one of the most serious nonnative invasive forest insects discovered in North America in recent years. A. glabripennis is regulated by federal quarantines in the United States and Canada and is the subject of eradication programs that involve locating, cutting, and chipping all infested trees. Other control methods are needed to aid in eradication and to form an integrated management program in the event eradication fails. We conducted laboratory bioassays to determine the toxicity of two systemic insecticides, azadirachtin and imidacloprid, for potential control of A. glabripennis and the cottonwood borer, Plectrodera scalator (F.) (Coleoptera: Cerambycidae), a closely related native cerambycid. Larvae of both cerambycid species were fed artificial diet with dilutions of azadirachtin or imidacloprid for 14 wk. Both insecticides exhibited strong antifeedant effects and some toxicity against A. glabripennis and P. scalator larvae. For A. glabripennis, the highest larval mortality at the end of the bioassay was 60% for larvae fed artificial diet treated with azadirachtin (50 ppm) or imidacloprid (1.6 ppm). For P. scalator, the highest larval mortality at the end of the bioassay was 100% for larvae fed artificial diet treated with azadirachtin (50 ppm) or imidacloprid (160 ppm). At 14 wk, the LC50 values for P. scalator were 1.58 and 1.78 ppm for azadirachtin and imidacloprid, respectively. Larvae of both species gained weight when fed diet treated with formulation blanks (inert ingredients) or the water control but lost weight when fed diet treated with increasing concentrations of either azadirachtin or imidacloprid. In a separate experiment, A. glabripennis adults were fed maple twigs treated with high and low concentrations of imidacloprid. A. glabripennis adult mortality reached 100% after 13 d on twigs treated with 150 ppm imidacloprid and after 20 d on twigs treated with 15 ppm imidacloprid. There was no visible feeding by A. glabripennis adults on twigs treated at the higher imidacloprid rate, and feeding was significantly reduced for adults placed on twigs treated at the low imidacloprid rate compared with adults on untreated twigs. In summary, imidacloprid and azadirachtin had both antifeedant and toxic effects against A. glabripennis and P. scalator and have potential for use in management programs. Based on our results, the delivery of high and sustained insecticide concentrations will be needed to overcome the antifeedant effects and lengthy lethal time for both larvae and adults exposed to these insecticides.


Determination of Optimal Heart Rate Variability Features Based on SVM-Recursive Feature Elimination for Cumulative Stress Monitoring Using ECG Sensor.
Routine stress monitoring in daily life can predict potentially serious health impacts. Effective stress monitoring in medical and healthcare fields is dependent upon accurate determination of stress-related features. In this study, we determined the optimal stress-related features for effective monitoring of cumulative stress. We first investigated the effects of short- and long-term stress on various heart rate variability (HRV) features using a rodent model. Subsequently, we determined an optimal HRV feature set using support vector machine-recursive feature elimination (SVM-RFE). Experimental results indicate that the HRV time domain features generally decrease under long-term stress, and the HRV frequency domain features have substantially significant differences under short-term stress. Further, an SVM classifier with a radial basis function kernel proved most accurate (93.11%) when using an optimal HRV feature set comprising the mean of R-R intervals (mRR), the standard deviation of R-R intervals (SDRR), and the coefficient of variance of R-R intervals (CVRR) as time domain features, and the normalized low frequency (nLF) and the normalized high frequency (nHF) as frequency domain features. Our findings indicate that the optimal HRV features identified in this study can effectively and efficiently detect stress. This knowledge facilitates development of in-facility and mobile healthcare system designs to support stress monitoring in daily life.


Current decision-making in prostate cancer therapy.
Prostate cancer continues to be the most prevalent cancer among American men. Localized prostate cancer is commonly diagnosed because of improved screening practices nationwide. Several options exist for the treatment of localized prostate cancer, and this review discusses the decision-making process facing patients diagnosed with this disease. No one treatment for localized prostate cancer has proven superior to date. For this reason patients have been found to use a number of resources to make an informed decision. These include physicians, spouses, family, friends, and different media. Urologists serve as the primary and most influential physicians and play an important role in the decision-making process. Patients, however, are assuming a more active role in this process as time evolves, especially with ease of access to multiple information resources. In deciding on a treatment for localized prostate cancer, patients must weigh the risks and benefits of each option. Urologists must provide patients with up-to-date information on these options and be aware of the different influences that surround these men during the decision-making process.


The proteasome as a potential target for novel anticancer drugs and chemosensitizers.
A major challenge in cancer therapy is tumor drug resistance. To overcome it, it is essential to understand the mechanisms and identify the molecules involved, so that they can be specifically targeted in combination therapies. The proteasome is such a validated target: it plays a key role in cancer cell proliferation, inhibition of chemotherapy-induced apoptosis and drug resistance development. Bortezomib (Velcade, PS-341) was the first proteasome inhibitor to receive regulatory approval from the US Food and Drug Administration for the treatment of multiple myeloma. Clinical combination trials have demonstrated a chemo-sensitizing effect of bortezomib on conventional agents in hematological malignancies and some solid tumors such as androgen-independent prostate and ovarian cancer. Although generally well-tolerated, bortezomib still generates toxicity which underscores the need for less toxic proteasome inhibitors. Several naturally occurring products, such as green tea polyphenols and the antibiotic lactacystin, have been shown to be potent proteasome inhibitors. Significantly, green tea polyphenols, as well as several flavonoids such as genistein, curcumin and resveratrol, have also been shown to have chemo-sensitizing properties in prostate, breast, hepatic, and lung tumors. Further studies on natural proteasome inhibitors as chemo-sensitizers could lead to identification of more potent and less toxic compounds that could be used in combination therapies for drug-resistant tumors.


Antimicrobial resistance in Swiss laying hens, prevalence and risk factors.
Antimicrobial resistance is an emerging concern to public health, and food-producing animals are known to be a potential source for transmission of resistant bacteria to humans. As legislation of the European Union requires to ban conventional cages for the housing of laying hens on the one hand, and a high food safety standard for eggs on the other hand, further investigations about the occurrence of antimicrobial resistance in alternative housing types are required. In this study, we determined antimicrobial resistance in indicator bacteria from 396 cloacal swabs from 99 Swiss laying hen farms among four alternative housing types during a cross-sectional study. On each farm, four hens were sampled and exposure to potential risk factors was identified with a questionnaire. The minimal inhibitory concentration was determined using broth microdilution in Escherichia coli (n=371) for 18 antimicrobials and in Enterococcus faecalis (n=138) and Enterococcus faecium (n=153) for 16 antimicrobials. All antimicrobial classes recommended by the European Food Safety Authority for E. coli and enterococci were included in the resistance profile. Sixty per cent of the E. coli isolates were susceptible to all of the considered antimicrobials and 30% were resistant to at least two antimicrobials. In E. faecalis, 33% of the strains were susceptible to all tested antimicrobials and 40% were resistant to two or more antimicrobials, whereas in E. faecium these figures were 14% and 39% respectively. Risk factor analyses were carried out for bacteria species and antimicrobials with a prevalence of resistance between 15% and 85%. In these analyses, none of the considered housing and management factors showed a consistent association with the prevalence of resistance for more than two combinations of bacteria and antimicrobial. Therefore we conclude that the impact of the considered housing and management practices on the egg producing farms on resistance in laying hens is low.


Benchmarking multiple fragmentation methods on an orbitrap fusion for top-down phospho-proteoform characterization.
Top-down analysis of intact proteins by mass spectrometry provides an ideal platform for comprehensive proteoform characterization, in particular, for the identification and localization of post-translational modifications (PTM) co-occurring on a protein. One of the main bottlenecks in top-down proteomics is insufficient protein sequence coverage caused by incomplete protein fragmentation. Based on previous work on peptides, increasing sequence coverage and PTM localization by combining sequential ETD and HCD fragmentation in a single fragmentation event, we hypothesized that protein sequence coverage and phospho-proteoform characterization could be equally improved by this new dual fragmentation method termed EThcD, recently been made available on the Orbitrap Fusion. Here, we systematically benchmark the performance of several (hybrid) fragmentation methods for intact protein analysis on an Orbitrap Fusion, using as a model system a 17.5 kDa N-terminal fragment of the mitotic regulator Bora. During cell division Bora becomes multiply phosphorylated by a variety of cell cycle kinases, including Aurora A and Plk1, albeit at distinctive sites. Here, we monitor the phosphorylation of Bora by Aurora A and Plk1, analyzing the generated distinctive phospho-proteoforms by top-down fragmentation. We show that EThcD and ETciD on a Fusion are feasible and capable of providing richer fragmentation spectra compared to HCD or ETD alone, increasing protein sequence coverage, and thereby facilitating phosphosite localization and the determination of kinase specific phosphorylation sites in these phospho-proteoforms. Data are available via ProteomeXchange with identifier PXD001845.


On localization for double Fourier series.
The localization theorems for Fourier series of functions of a single variable are classical and easy to prove. The situation is different for Fourier series of functions of several variables, even if one restricts consideration to rectangular, in particular square, partial sums. We show that the answer to the problem can be obtained by considering the notion of generalized bounded variation, which we introduced. Given a nondecreasing sequence {lambda(n)} of positive numbers such that Sigma 1/lambda(n) diverges, a function g defined on an interval I of R(1) is said to be of Lambda-bounded variation (LambdaBV) if Sigma|g(a(n)) - g(b(n))|/lambda(n) converges for every sequence of nonoverlapping intervals (a(n), b(n)) [unk]I. If lambda(n) = n, we say that g is of harmonic bounded variation (HBV). The definition suitably modified can be extended to functions of several variables. We show that in the case of two variables the localization principle holds for rectangular partial sums if LambdaBV = HBV, and that if LambdaBV is not contained in HBV, then the localization principle does not hold for LambdaBV even in the case of square partial sums.


In vitro activity of bergamot natural essence and furocoumarin-free and distilled extracts, and their associations with boric acid, against clinical yeast isolates.
There is very little information, to date, on the antifungal activity of bergamot oil. In this study, we investigated the in vitro activity of three bergamot oils (natural essence, furocoumarin-free extract and distilled extract) against clinically relevant Candida species. We studied the two derivatives, components of Italian pharmaceutical products, that are supposed to be less toxic than the essential oil. In vitro susceptibility of 40 clinical isolates of Candida spp. (Candida albicans, n=20; Candida glabrata, n=13; Candida krusei, n=4; Candida tropicalis, n=2; Candida parapsilosis, n=1), associated with symptomatic and asymptomatic vulvovaginal candidiasis, was determined using a modification of the NCCLS M27-A2 broth microdilution method. MICs were evaluated for each of the oils alone and combined with sub-inhibitory concentrations of the well-known antiseptic, boric acid. To boric acid, all isolates had MIC values ranging from 0.094% to 0.187% (w/v). At 24 h readings, the MIC(90 )s (for all isolates) were (v/v): 5% for natural essence of bergamot, 2.5% for the furocoumarin-free extract, and 1.25% for the distilled extract. At the 48 h reading, these values increased to >10%, 5% and 2.5%, respectively. At both readings, MIC(90 )s for all oil+boric acid combinations were significantly lower than corresponding values for the oils alone (P <0.05). These data indicate that bergamot oils are active in vitro against Candida spp., suggesting their potential role for the topical treatment of Candida infections.


Development of fetal haemoglobin-blood cells (F cells) within colorectal tumour tissues.
To evaluate the sources of fetal haemoglobin (HbF) as an indicator in cancer. An immunohistochemical study was carried out on some of the most common kinds of cancer. All of these cancers had serologically high levels of HbF as evaluated previously. Immunoaffinity-purified anti-HbF was immunohistochemically used to study F cell distribution in the following cancers: colorectal adenocarcinoma, urinary bladder transitional cell carcinoma, brain tumours, lung carcinoma, breast adenocarcinoma, leukaemia, Burkitt's lymphoma and endometrial carcinoma. In colorectal adenocarcinoma, HbF-containing red blood cells (FRBC) were present within thin-walled vessels or were disposed in dense clusters within the tumour. Some of these cells were nucleated or binucleated HbF-erythroblasts or HbF-normoblasts (FNBS). In two cases, high levels of mitoses within HbF-erythroblasts were observed. In half of the cases with transitional cell carcinoma of the urinary bladder, regional intratumoral blood vessels were found to contain 5-50% FRBC. In the other tumours examined, F cells were not observed. FRBCs, however, were occasionally observed in the regional lymph nodes of some of these cancers. The evaluation of HbF as a potential plasma marker is suggested by the high concentration of FRBCs in colorectal tumours. The apparent development of FRBCs in colorectal tumour tissues is an interesting observation, as these cells were previously thought to develop in medullary or lymphoid tissues. It is thus suggested that the colonic microenvironment may stimulate extramedullary fetal-type haematopoiesis.


Intraarticular Injections of Platelet-rich Plasma (PRP) in the Management of Knee Osteoarthritis.
The clinical use of PRP therapy in the practical setting of orthopaedic fields is increasing partly because of the accessibility of devices that are used in outpatient preparation and delivery. Another reason is the strong advertisement of PRP procedures as the ultimate treatment and novel technology for knee problems by a few orthopaedic surgeons based on claims of abundant scientific evidence. Hence, PubMed articles related to the clinical use of PRP in knee osteoarthritis were searched using the key words: PRP, knee and osteoarthritis in order to study these claims. A total of 20 reports were found directly related to the topic. The aforementioned clinical studies suggest that intraarticular injections of PRP could have preventive effects against osteoarthritis progression. However, presently there is no clear evidence from well-designed clinical trials that intraarticular injections of PRP are efficacious in osteoarthritis. Therefore, at this time the efficacy of PRP requires more investigation, wherein better scientific studies should be performed that include high powered randomized controlled trials.


Calcium overload in human giant cell myocarditis.
Myocardial calcium overload was observed in a patient with giant cell myocarditis. The myocardial calcium content estimated by atomic absorption spectrophotometry amounted to 120 mEq/kg dry weight, and the von Kossa stain disclosed multiple foci with patchy calcifications of myocardial fibres. Cytochemical examination of the ultrastructural calcium localisation using the phosphate-pyroantimonate method showed considerable variation in the subcellular calcium distribution. In normal myocytes calcium precipitates were confined to the inner leaflet of the sarcolemma, T-tubules, intercalated disks, and sporadically to mitochondria. In contrast, extensive calcification of mitochondria and loss of sarcolemmal calcium was evident in necrotic myocytes. A number of grossly normal myocytes also showed an increase of calcium precipitates in slightly swollen mitochondria. These findings suggest that myocardial calcium overload in this case started in viable myocytes and was not merely a secondary phenomenon occurring after cell death.


New approaches for the detection of minimal residual disease in acute myeloid leukemia.
The detection of minimal residual disease (MRD) in patients with acute leukemia has been studied for about 15 years by different groups in both the United States and Europe. It has been found that MRD detection can be performed using molecular and immunophenotypic aberrancies that are present in the leukemic clone at diagnosis and not in normal bone marrow. When performing MRD assessments after chemotherapy, it is possible to identify patients at risk for relapse. This review is not an overview of all MRD studies, but rather discusses the possibilities for optimizing MRD detection, the use of flow cytometry versus polymerase chain reaction techniques, and the implications for future patient treatment. When informative, we compare literature on MRD in acute myeloid leukemia (AML) with information from MRD studies in acute lymphoblastic leukemia. Finally, we address the promising detection of AML stem cells, the likely cells of origin in AML, for prediction of clinical outcome and guidance of future therapies.


Taking Inventory and Moving Forward: A Review of the Research Literature and Assessment of Qualitative Research in JPCC, 2010-2014.
As the foremost journal in spiritual care and counseling (SCC), Journal of Pastoral Care & Counseling (JPCC) functions as a barometer for the discipline's research and interests. This article presents the findings of a review of the research literature in JPCC between 2010 and 2014. It examines research articles by asking the following questions: What are the quantity and types of research published? What are the dominant themes in this research? What are the quantity and methodologies of qualitative research? Findings are presented, discussed and recommendations are made in an effort to assess and further build the research base of the discipline.


Long-term outcomes in pediatric liver recipients: comparison between cyclosporin A and tacrolimus.
In recent years, tacrolimus (FK506, TAC) has been increasingly utilized in liver transplantation. However, long-term risks and benefits as compared with conventional cyclosporin A (CsA) have not been fully elucidated. This retrospective study examined the potential outcome differences between TAC- and CsA-based immunosuppressive therapy in pediatric liver transplant recipients. From March 1988 to December 1996, 218 children (aged 0.1-17 yr) underwent 238 orthotopic liver transplantations; 58.7% (128/218) were under 2 yr of age at time of transplant. Initially, the maintenance immunosuppressive regimen consisted of CsA and prednisone, with antilymphocytic preparations (MALG, ATGAM, and OKT3) as induction therapy. Subsequently, TAC was used first as rescue therapy for steroid refractory rejection in CsA patients and then as maintenance immunosuppression. Fifty-seven out of the 147 CsA patients were converted to TAC for various reasons while 71 patients were placed on TAC as primary maintenance immunosuppression. 62.6 per cent (92/147) of liver recipients on CsA experienced at least one biopsy-proven acute rejection episode as compared to 50.7% (36/71) for TAC patients (p = 0.09); likewise, 34% (50/147) of CsA patients had more than one episode of rejection vs. 18.3% (13/71) for patients on TAC (p < 0.02). Rejection was the reason for conversion from CsA to TAC in 29 of 57 patients. Conversely, 19.0% (28/147) of CsA patients had to be switched to TAC for reasons not related to rejection (i.e. side-effects). The overall incidence of histologically proven chronic rejection was 7.8% (17/218). 10.9 per cent (16/147) of the children who were on CsA initially developed chronic rejection, which was significantly higher compared with one of 71 TAC recipients (p < 0.02). Of these 16 CsA patients with chronic rejection, 50.0% (8/16) underwent retransplantation for graft failure (mean interval from time of diagnosis of chronic rejection to re-transplant, 4.0 months; range 1-8 months), whereas the TAC patient has remained clinically stable with normal liver function tests after 23 months of follow-up. One year after liver transplantation, 72.8% (107/147) of CsA patients were still on steroids (mean dosage 0.20 mg/kg/d), as compared to 42.3% (30/71) of the TAC patients (mean dosage 0.14 mg/kg/d). The incidence of post-transplant lymphoproliferative disorder (PTLD) in Epstein-Barr virus (EBV)-infected patients was 2.2% (2/90), 7.0% (5/71) and 12.3% (7/57) for CsA, primary and TAC-converted groups, respectively. The overall incidence of PTLD was 6.9% (15/218). In summary, pediatric liver transplant recipients treated with TAC as primary maintenance immunosuppressive medication experienced significantly fewer episodes of rejection; especially chronic rejection, which lead to graft loss. However, the trade-off is a potential increased incidence of EBV-related PTLD in these patients.


Root hair development in grasses and cereals (Poaceae).
Root hairs are tubular, cellular outgrowths of epidermal cells that extend from the root surface into the soil. Root hairs tether root systems to their growth substrate, take up inorganic nutrients and water, and interact with the soil microflora. At maturity, the root epidermis comprises two cell types; cells with root hairs and hairless epidermal cells. These two cell types alternate with each other along longitudinal files in grasses and cereals (Poaceae). While the mechanism by which this alternating pattern develops is unknown, the later stages of root hair differentiation are controlled by a conserved mechanism that promotes root hair development among angiosperms.


Three dimensional echo-planar imaging at 7 Tesla.
Functional MRI (fMRI) most commonly employs 2D echo-planar imaging (EPI). The advantages for fMRI brought about by the increasingly popular ultra-high field strengths are best exploited in high-resolution acquisitions, but here 2D EPI becomes impractical for several reasons, including the very long volume acquisitions times. In this study at 7 T, a 3D EPI sequence with full parallel and partial Fourier imaging capability along both phase encoding axes was implemented and used to evaluate the sensitivity of 3D and corresponding 2D EPI acquisitions at four different spatial resolutions ranging from small to typical voxel sizes (1.5-3.0 mm isotropic). Whole-brain resting state measurements (N=4) revealed a better, or at least comparable sensitivity of the 3D method for gray and white matter. The larger vulnerability of 3D to physiological effects was outweighed by the much shorter volume TR, which moreover allows whole-brain coverage at high resolution within fully acceptable limits for event-related fMRI: TR was only 3.07 s for 1.5 mm, 1.88 s for 2.0 mm, 1.38 s for 2.5 mm and 1.07 s for 3.0 mm isotropic resolution. In order to investigate the ability to detect and spatially resolve BOLD activation in the visual cortex, functional 3D EPI experiments (N=8) were performed at 1 mm isotropic resolution with parallel imaging acceleration of 3x3, resulting in a TR of only 3.2 s for whole-brain coverage. From our results, and several other practical advantages of 3D over 2D EPI found in the present study, we conclude that 3D EPI provides a useful alternative for whole-brain fMRI at 7 T, not only when high-resolution data are required.


Sensing Ability and Formation Criterion of Fluid Supported Lipid Bilayer Coated Graphene Field-Effect Transistors.
Supported lipid bilayers (SLBs) have been widely used to provide native environments for membrane protein studies. In this study, we utilized graphene field-effect transistors (GFETs) coated with a fluid SLB to perform label-free detection of membrane-associated ligand-receptor interactions in their native lipid bilayer environment. It is known that the analyte-binding event needs to occur within the Debye length for it to be significantly sensed by an FET sensor. However, the thickness of a lipid bilayer is around 4-5-nm-thick, which is larger than the Debye length of a solution with physiologically relevant ionic strength. There is thus a question of whether an FET sensor can detect the binding event above the bilayer. In this study, we show how the existence of an SLB can influence the effective detection distance and the formation criterion of a fluid and continuous SLB on a graphene surface. We discovered that the water intercalation between the graphene and the underlying silica substrate hinders the SLB formation but is required for the stable electrical recording by a GFET. To verify the existence of a fluid SLB on graphene, which was previously complicated by the graphene fluorescence quenching effect, we developed a modified fluorescence recovery after photobleaching method. In addition, our results showed that SLB coated GFETs can quantitatively detect ligand binding onto the receptors embedded in the SLBs. The comparison of our experimental data with a theoretical model shows that the contribution of the SLB acyl chain hydrophobic region to the screening effect can be negligible and, therefore, that the effective detection region can extend beyond the SLB.


Hyperparasitism of the cryptoniscid isopod Liriopsis pygmaea on the lithodid Paralomis granulosa from the Beagle Channel, Argentina.
A total of 29,570 false king crab Paralomis granulosa were sampled from the Beagle Channel (54 degrees 51'S, 68 degrees 12'W), Tierra del Fuego, Argentina, between July 1996 and August 1998. Crab size varied from 6.8 to 111.2 mm carapace length (CL). A few crabs parasitized by the rhizocephalan Briarosaccus callosus were found; prevalences of externae (the rhizocephalan reproductive body) and scars (the mark left on the host after the death of the parasite) were 0.28 and 0.16%, respectively. Of 85 externae examined, 55 were non-ovigerous and 30 ovigerous. The cryptoniscid isopod Liriopsis pygmaea infested 36.5% of the B. callosus examined. The most abundant stage was the cryptonicus larva, accounting for 208 of the 238 L. pygmaea recovered. Cryptonisci showed a highly aggregated distribution. A total of 92.7% of cryptonicsci were recovered inside empty externae, suggesting that the latter were attractive to cryptonisci. Early subadult females of L. pygmaea were rare; only 3 individuals occurred inside 1 ovigerous externa. Eight late subadult and 18 adult females were found on 3 and 7 non-ovigerous externae, respectively; in addition, 1 aberrant late subadult was found on 1 ovigerous externa. In the Beagle Channel, the population of P. granulosa harbours 3 different parasites: the bopyrid isopod Pseudione tuberculata, which reaches highest prevalence at 10 to 20 mm CL, the rhizocephalan B. callosus, with highest prevalence at 20 to 40 mm CL, and the cryptoniscid isopod L. pygmaea, which mainly infests rhizocephalan on crabs >40 mm CL.


A review of state regulations to promote physical activity and safety on playgrounds in child care centers and family child care homes.
As interventions increasingly emphasize early child care settings, it is necessary to understand the state regulatory context that provides guidelines for outdoor physical activity and safety and sets standards for child care environments. Researchers reviewed regulations for child care facilities for 50 states, the District of Columbia and the Virgin Islands. We compared state regulations with national standards for 17 physical activity- and safety-related items for outdoor playground settings outlined in Caring for Our Children: National Health and Safety Performance Standards: Guidelines for Out-of-Home Child Care Programs (CFOC). State regulations were coded as fully, partially or not addressing the CFOC standard and state-level summary scores were calculated. On average, state regulations fully addressed one-third of 17 CFOC standards in regulations for centers (34%) and family child care homes (27%). Data suggest insufficient attention to outdoor play area proximity and size, equipment height, surfacing, and inspections. Considerable variation exists among state regulations related to physical activity promotion and injury prevention within outdoor play areas. Many states' regulations do not comply with published national health and safety standards. Enhancing regulations is one component of a policy approach to promoting safe, physically active child care settings.


Pornography use and closeness with others in men.
The aim of the present study was to examine the association between pornography use and aspects of socioemotional closeness in a non-clinical sample of 164 males. Participants' actual and ideal socioemotional closeness was measured by the Perceived Interpersonal Closeness Scale (PICS; Popovic et al., 2003) while their pornography use was examined by the Background and Pornography Use Information Questionnaire (Popovic, 2009). Potential effects of these variables on each other as well as various findings concerning pornography consumption and related attitudes are presented. The results showed that there was no significant difference between self-reported pornography users and non-users in terms of specific socioemotional closeness with the most significant adults in their lives (i.e., partners, closest friends, mothers, and fathers). However, pornography users had significantly higher total closeness scores than non-users, showing possibly a craving for intimacy among pornography users versus non-pornography users. This "intimacy motive" and relevant and controversial findings are discussed.


Polarization properties of a grating spectrograph.
The 13.7-m Czerny-Turner spectrograph at the McMath solar telescope is evaluated with regard to polarization properties. Let I || = the transmissivity of the spectrograph for incident light linear polarized parallel to the entrance slit and I perpendicular = transmissivity for light polarized perpendicular to the entrance slit. The ratio I ||/I perpendicular was measured photoelectrically as a function of wavelength in six diffraction orders. Values of this ratio vary from 0.3 to 20. It is shown that spectrograph transmission as a function of polarization may cause large photometric and radiometric errors. This fact is not unique to this particular instrument. Two mechanisms appear to contribute to the polarizance of the grating. These are: (1) the Rayleigh or Wood's anomalies in which polarizance maxima and minima occur and (2) a vector wave interaction introduced because groove dimension is nearly the same as the wavelength. For lambda > groove depth it is found that the wavelength of a peak polarizance (lambda(p)) is given by lambda(p) = 0.7d costheta, where d is the ruling separation and theta is the grating angle. Photoelectric scans of the solar spectrum were made across several of the Wood's anomalies to obtain their profiles.


Mortality due to bleeding, myocardial infarction and stroke in dialysis patients.
Essentials Mortality due to bleeding vs. arterial thrombosis in dialysis patients is unknown. We compared death causes of 201 918 dialysis patients with the general population. Dialysis was associated with increased mortality risks of bleeding and arterial thrombosis. Clinicians should be aware of the increased bleeding and thrombosis risks. Background Dialysis has been associated with both bleeding and thrombotic events. However, there is limited information on bleeding as a cause of death versus arterial thrombosis as a cause of death. Objectives To investigate the occurrence of bleeding, myocardial infarction and stroke as causes of death in the dialysis population as compared with the general population. Methods We included 201 918 patients from 11 countries providing data to the ERA-EDTA Registry who started dialysis treatment between 1994 and 2011, and followed them for 3 years. Age-standardized and sex-standardized mortality rate ratios for bleeding, myocardial infarction and stroke as causes of death were calculated in dialysis patients as compared with the European general population. Associations between potential risk factors and these causes of death in dialysis patients were investigated by calculating hazard ratios (HRs) with 95% confidence intervals (CIs) by the use of Cox proportional-hazards regression. Results As compared with the general population, the age-standardized and sex-standardized mortality rate ratios in dialysis patients were 12.8 (95% CI 11.9-13.7) for bleeding as a cause of death (6.2 per 1000 person-years among dialysis patients versus 0.3 per 1000 person-years in the general population), 13.4 (95% CI 13.0-13.9) for myocardial infarction (22.5 versus 0.9 per 1000 person-years), and 12.4 (95% CI 11.9-12.9) for stroke (14.3 versus 0.7 per 1000 person-years). Conclusion Dialysis patients have highly increased risks of death caused by bleeding and arterial thrombosis as compared with the general population. Clinicians should be aware of the increased mortality risks caused by these conditions.


Insensitivity to the Fungicide Fosetyl-Aluminum in California Isolates of the Lettuce Downy Mildew Pathogen, Bremia lactucae.
Lettuce downy mildew, caused by Bremia lactucae, is the most important foliar disease of lettuce in California. In recent years, there were apparent failures of fungicides containing fosetyl-aluminum (Aliette) to control downy mildew in commercial lettuce fields in California. Consequently, we characterized 134 isolates collected over 2 years from throughout the coastal growing areas of California for insensitivity to the fungicides fosetyl-aluminum and maneb, pathotype, and mating type. Tests using seedlings in controlled growth room conditions demonstrated the widespread occurrence of insensitivity to fosetyl-aluminum in California populations of B. lactucae. Fifty percent of the isolates assayed sporulated profusely in the presence of fosetyl-aluminum applied at rates twice the normal field dosage, and an additional 40% showed moderate sporulation at this rate. Fosetyl-aluminum-insensitive isolates were detected from all regions sampled. Insensitivity was also observed in multiple pathotypes. Insensitivity was not complete, however, because quantitative analysis of the number of lesions on older plants revealed that applications of fosetyl-aluminum could reduce the levels of disease by 50%. Therefore, while fosetyl-aluminum may have utility under low disease pressure in the field, other control measures are required to provide control under conditions favorable to the disease.


Review of Mathematical Models of Vaccination for Preventing Congenital Cytomegalovirus Infection.
Several cytomegalovirus (CMV) vaccine candidates are under development. To reduce the burden of congenital CMV infection, potential strategies under consideration include vaccination of adult women, adolescent girls, and/or young children (both sexes). We reviewed 5 studies that used infectious disease modeling to assess the potential impact of vaccination for preventing congenital CMV infection. All models assumed CMV vaccination would prevent primary infection and 2 models also assumed prevention of reinfections and reactivations. Despite differences in structure, assumptions, and population data, infant vaccination (both sexes) was the optimal strategy in all models, but in 1 model vaccinating seronegative women at 19-21 years of age was also optimal (for duration of vaccine protection ≥8 years). In 3 models, infant vaccination increased average age at primary infection as a result of decreased secondary transmission (herd immunity) combined with waning vaccine-induced immunity. This effect could increase the risk of congenital CMV infections in populations where primary CMV infection occurs early in childhood but could be minimized by administering a second dose of vaccine during adolescence. Understanding vaccine efficacy and duration of immunity, and how these might vary depending on CMV serostatus and age at vaccination, will be key to defining CMV vaccination strategies.


Subpopulation-specific differences in skeletal muscle mitochondria in humans with obesity: insights from studies employing acute nutritional and exercise stimuli.
Mitochondria from skeletal muscle of humans with obesity often display alterations with respect to their morphology, proteome, biogenesis, and function. These changes in muscle mitochondria are considered to contribute to metabolic abnormalities observed in humans with obesity. Most of the evidence describing alterations in muscle mitochondria in humans with obesity, however, lacks reference to a specific subcellular location. This is despite data over the years showing differences in the morphology and function of subsarcolemmal (found near the plasma membrane) and intermyofibrillar (nested between the myofibrils) mitochondria in skeletal muscle. Recent studies reveal that impairments in mitochondrial function in obesity with respect to the subcellular location of the mitochondria in muscle are more readily evident following exposure of the skeletal muscle to physiological stimuli. In this review, we highlight the need to understand skeletal muscle mitochondria metabolism in obesity in a subpopulation-specific manner and in the presence of physiological stimuli that modify mitochondrial function in vivo. Experimental approaches employed under these conditions will allow for more precise characterization of impairments in skeletal muscle mitochondria and their implications in inducing metabolic dysfunction in human obesity.


Outcomes of transcatheter mitral valve replacement for degenerated bioprostheses, failed annuloplasty rings, and mitral annular calcification.
We sought to evaluate the outcomes of transcatheter mitral valve replacement (TMVR) for patients with degenerated bioprostheses [valve-in-valve (ViV)], failed annuloplasty rings [valve-in-ring (ViR)], and severe mitral annular calcification [valve-in-mitral annular calcification (ViMAC)]. From the TMVR multicentre registry, procedural and clinical outcomes of ViV, ViR, and ViMAC were compared according to Mitral Valve Academic Research Consortium (MVARC) criteria. A total of 521 patients with mean Society of Thoracic Surgeons score of 9.0 ± 7.0% underwent TMVR (322 patients with ViV, 141 with ViR, and 58 with ViMAC). Trans-septal access and the Sapien valves were used in 39.5% and 90.0%, respectively. Overall technical success was excellent at 87.1%. However, left ventricular outflow tract obstruction occurred more frequently after ViMAC compared with ViR and ViV (39.7% vs. 5.0% vs. 2.2%; P < 0.001), whereas second valve implantation was more frequent in ViR compared with ViMAC and ViV (12.1% vs. 5.2% vs. 2.5%; P < 0.001). Accordingly, technical success rate was higher after ViV compared with ViR and ViMAC (94.4% vs. 80.9% vs. 62.1%; P < 0.001). Compared with ViMAC and ViV groups, ViR group had more frequent post-procedural mitral regurgitation ≥moderate (18.4% vs. 13.8% vs. 5.6%; P < 0.001) and subsequent paravalvular leak closure (7.8% vs. 0.0% vs. 2.2%; P = 0.006). All-cause mortality was higher after ViMAC compared with ViR and ViV at 30 days (34.5% vs. 9.9% vs. 6.2%; log-rank P < 0.001) and 1 year (62.8% vs. 30.6% vs. 14.0%; log-rank P < 0.001). On multivariable analysis, patients with failed annuloplasty rings and severe MAC were at increased risk of mortality after TMVR [ViR vs. ViV, hazard ratio (HR) 1.99, 95% confidence interval (CI) 1.27-3.12; P = 0.003; ViMAC vs. ViV, HR 5.29, 95% CI 3.29-8.51; P < 0.001]. The TMVR provided excellent outcomes for patients with degenerated bioprostheses despite high surgical risk. However, ViR and ViMAC were associated with higher rates of adverse events and mid-term mortality compared with ViV.


Generation and characterization of dipeptidyl peptidase-IV inhibitory peptides from trypsin-hydrolyzed α-lactalbumin-rich whey proteins.
Dipeptidyl peptidase-IV (DPP-IV) is an enzyme that break down the antidiabetic hormone glucagon-like peptide-1. Therefore, inhibition of DPP-IV could be an effective strategy to treat Type 2 diabetes (T2D). The α-lactalbumin-rich whey protein concentrate was hydrolyzed by trypsin, and the hydrolysates were then fractionated at a semi-preparative scale using a Superdex Gel filtration Chromatography. The peptides were analyzed by using HPLC coupled with tandem mass spectrometry (RP-HPLC-MS/MS), and their Dipeptidyl peptidase-IV inhibitory activity was determined by the enzymatic assay. Among tested fragments, a potent fragment (LDQWLCEKL), with the half-maximal inhibitory concentration (IC50) of 131 μM was obtained. Further analysis shows that the LDQWLCEKL peptide corresponds to the amino acid sequence of f(115-123) in α-lactalbumin. Furthermore, LDQWLCEKL exhibited a typical non-competitive mode of inhibition. The results indicate that α-lactalbumin contains active peptides with DPP-IV inhibitory activity that may be used to prevent and treat T2D.