Exosomes Isolated from Ascites of T-Cell Lymphoma-Bearing Mice Expressing Surface CD24 and HSP-90 Induce a Tumor-Specific Immune Response.
Extracellular vesicles (EVs), including endosome-derived nanovesicles (exosomes), are involved in cell-cell communication. Through transfer of their molecular contents, extracellular nanovesicles can alter the function of recipient cells. Due to these characteristics, EVs have shown potential as a new alternative for cancer immunotherapy. Tumor exosomes isolated from malignant ascites can activate dendritic cells, thereby priming the immune system to recognize and kill cancer cells. However, a suppressive role on tumor immune response has also been reported, suggesting that the neoplastic stage of carcinogenesis and the microenvironment where tumor cells grow may influence the amount of EVs released by the cell. This neoplastic stage and microenvironment may also impact EVs' components such as proteins and miRNA, determining their biological behavior. Most T-cell lymphomas have an aggressive clinical course and poor prognosis. Consequently, complementary alternative therapies are needed to improve the survival rates achieved with conventional treatments. In this work, we have characterized EVs isolated from ascites of mice bearing a very aggressive murine T-cell lymphoma and have studied their immunogenic properties. Small EVs were isolated by differential centrifugation, ultrafiltration, and ultracentrifugation at 100,000 × g on a sucrose cushion. The EVs were defined as exosomes by their morphology and size analyzed by electron microscopy, their floating density on a sucrose gradient, as well as their expression of endosome marker proteins ALIX, TSG-101; the tetraspanins CD63, CD9, and CD81. In addition, they contain tumor antigens, the marker for malignancy CD24, the heat shock protein HSP-70, and an unusual surface expression of HSP-90 was demonstrated. The administration of EVs isolated from ascites (EVs A) into naïve-syngeneic mice induced both humoral and cellular immune responses that allowed the rejection of subsequent tumor challenges. However, the immunization had no effect on a non-related mammary adenocarcinoma, demonstrating that the immune response elicited was specific and also it induced immune memory. In vitro analysis demonstrated that T-cells from EVs A-immunized mice secrete IFN-γ in response to tumor stimulation. Furthermore, tumor-specific CD4+ and CD8+ IFN-γ secreting cells could be efficiently expanded from mice immunized with EVs A, showing that a T helper 1 response is involved in tumor rejection. Our findings confirm exosomes as promising defined acellular tumor antigens for the development of an antitumor vaccine.


Mu-opioid receptor activation in live cells.
Interaction of the mu-opioid receptor (MOP) with selected ligands was investigated in live cells using advanced imaging by confocal laser scanning microscopy integrated with fluorescence correlation spectroscopy and fluorescence cross-correlation spectroscopy. In PC12 cells stably transformed to express the fluorescently labeled MOP-enhanced green fluorescent protein construct, two pools of MOP were identified that could be discriminated by differences in their lateral mobility in the cell membrane. The majority of MOP receptors (80+/-10%) were characterized by a diffusion coefficient D(MOP,1) = (4+/-2) x 10(-11) m(2) s(-1), compared with the slowly moving fraction, D(MOP,2) = (4+/-2) x 10(-12) m(2) s(-1). On stimulation with selected agonists ([D-Ala(2),N-MePhe(4),Gly-ol(5)]enkephalin, enkephalin-heptapeptide Tyr-Gly-Gly-Phe-Met-Arg-Phe, morphine, and methadone), surface density of the MOP decreased, whereas the lateral mobility increased. In contrast, antagonists (naloxone and naltrexone) "froze" the receptor in the membrane, i.e., increased MOP surface density and decreased lateral mobility. Agonist activation was also accompanied by pronounced changes in the dynamics of plasma membrane lipids, as revealed by the general lipid marker 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate dye. The results provide new information about MOP activation in live cells at the molecular level, with a special focus on the dynamics of the intricate interplay between this receptor and the surrounding lipids.


A High-Throughput Screening Assay to Identify Kidney Toxic Compounds.
Kidney toxicity due to drugs and chemicals poses a significant health burden for patients and a financial risk for pharmaceutical companies. However, currently no sensitive and high-throughput in vitro method exists for predictive nephrotoxicity assessment. Primary human proximal tubular epithelial cells (HPTECs) possess characteristics of differentiated epithelial cells, making them a desirable model to use in in vitro screening systems. Additionally, heme oxygenase 1 (HO-1) protein expression is upregulated as a protective mechanism during kidney toxicant-induced oxidative stress or inflammation in HPTECs and can therefore be used as a biomarker for nephrotoxicity. In this article, we describe two different methods to screen for HO-1 increase: A homogeneous time resolved fluorescence (HTRF) assay and an immunofluorescence assay. The latter provides lower throughput but higher sensitivity due to the combination of two readouts, HO-1 intensity and cell number. The methods described in the protocol are amendable for other cell types as well. © 2016 by John Wiley & Sons, Inc.


Cognitive dissonance induction in everyday life: An fMRI study.
This functional magnetic resonance imaging (fMRI) study explored the neural substrates of cognitive dissonance during dissonance "induction." A novel task was developed based on the results of a separate item selection study (n = 125). Items were designed to generate dissonance by prompting participants to reflect on everyday personal experiences that were inconsistent with values they had expressed support for. One experimental condition (dissonance) and three control conditions (justification, consonance, and non-self-related inconsistency) were used for comparison. Items of all four types were presented to each participant (n = 14) in a randomized design. The fMRI analysis used a whole-brain approach focusing on the moments dissonance was induced. Results showed that in comparison with the control conditions the dissonance experience led to higher levels of activation in several brain regions. Specifically dissonance was associated with increased neural activation in key brain regions including the anterior cingulate cortex (ACC), anterior insula, inferior frontal gyrus, and precuneus. This supports current perspectives that emphasize the role of anterior cingulate and insula in dissonance processing. Less extensive activation in the prefrontal cortex than in some previous studies is consistent with this study's emphasis on dissonance induction, rather than reduction. This article also contains a short review and comparison with other fMRI studies of cognitive dissonance.


Site-directed mutagenesis of the rat beta1-adrenoceptor. Involvement of Tyr356 (7.43) in (+/-)cyanopindolol but not (+/-)[125Iodo]cyanopindolol binding.
To determine the role played by Tyr(356 (7.43)) in the rat beta(1)-adrenoceptor in binding the antagonists (+/-)cyanopindolol (4-[3-(t-butylamino]-3-(2'-cyano-indoloxy)-2-propanolol) and its iodinated analogue (+/-)[(125)Iodo]cyanopindolol (1-(t-butylamino]-3-(2'-cyano-3'-iodo-indoloxy)-2-propanolol), Tyr(356 (7.43)) was mutated to either Phe or Ala and binding affinities determined for wild type and mutant rat beta(1)-adrenoceptors. Our results indicate that Tyr(356 (7.43)) is important for (+/-)cyanopindolol, but not (+/-)[(125)Iodo]cyanopindolol, binding and that (+/-)cyanopindolol adopts a "reverse" binding orientation whereas (+/-)[(125)Iodo]cyanopindolol cannot be accommodated in this binding mode. We define a "reverse" antagonist binding mode as one where the aryloxy moiety interacts with residues on transmembrane helices 1, 2, 3 and 7. The beta(1)-adrenoceptor site-directed mutagenesis results are the first to support a "reverse" antagonist binding orientation and the involvement of Tyr(356 (7.43)) in this binding mode.


The Ras/Raf signaling pathway is required for progression of mouse embryos through the two-cell stage.
We have used microinjection of antisense oligonucleotides, monoclonal antibody, and the dominant negative Ras N-17 mutant to interfere with Ras expression and function in mouse oocytes and early embryos. Microinjection of either ras antisense oligonucleotides or anti-Ras monoclonal antibody Y13-259 did not affect normal progression of oocytes through meiosis and arrest at metaphase II. However, microinjection of fertilized eggs with constructs expressing Ras N-17 inhibited subsequent development through the two-cell stage. The inhibitory effect of Ras N-17 was overcome by simultaneous injection of a plasmid expressing an active raf oncogene, indicating that it resulted from interference with the Ras/Raf signaling pathway. In contrast to the inhibition of two-cell embryo development resulting from microinjection of pronuclear stage eggs, microinjection of late two-cell embryos with Ras N-17 expression constructs did not affect subsequent cleavages and development to morulae and blastocysts. It thus appears that the Ras/Raf signaling pathway, presumably activated by autocrine growth factor stimulation, is specifically required at the two-cell stage, which is the time of transition between maternal and embryonic gene expression in mouse embryos.


Autonomously functioning thyroid nodules in a patient with a thyrotropin-secreting pituitary adenoma: possible cause--effect relationship.
A 51-year-old female patient with long-standing hyperthyroidism due to a thyrotropin-secreting pituitary adenoma is reported, who became thyrotoxic again shortly after successful pituitary surgery. Functional testing and scintigraphy suggested the diagnosis of autonomous functioning thyroid nodules, which was confirmed by pathological examination of the resected thyroid tissue. This is the first report revealing the transition from a pituitary-dependent to a thyroid-dependent hyperthyroidism. Autonomous functioning thyroid nodules are, however, considered an intrinsic thyroid defect. In similarity with other disorders, in which trophic hormones may induce an autonomous secretion by the target gland, this report opens the possibility that a humoral factor may play a role in the development of autonomous functioning thyroid nodules.


Luteinising hormone-based protocol versus traditional flexible gonadotropin-releasing hormone antagonist protocol in women with normal ovarian response: study protocol for a non-inferiority trial.
Many patients demonstrate an insufficient endogenous luteinising hormone (LH) concentration during ovarian stimulation. With traditional fixed or flexible gonadotropin-releasing hormone (GnRH) antagonist protocols, antagonist administration may further reduce LH activity. Previously, we proved that LH can be used as an indicator for the timing and dosage of antagonist. Patients with a persistently low LH concentration during ovarian stimulation may not require antagonists, whereas antagonist administration can affect reproductive outcomes. To further explore this hypothesis, we designed a randomised clinical trial to compare the LH-based flexible GnRH antagonist protocol with traditional flexible GnRH antagonist protocol in women with normal ovarian response. This study was a multicentre, parallel, prospective, randomised, non-inferiority study. The primary efficacy endpoint was cumulative ongoing pregnancy rate per cycle. The study aimed to prove the non-inferiority of cumulative ongoing pregnancy rate per cycle with an LH-based flexible GnRH antagonist protocol versus traditional flexible GnRH antagonist protocol. Secondary endpoints were the high-quality embryo rate, clinical pregnancy rate and cancellation rate. Differences in cost-effectiveness and adverse events were evaluated. The cumulative ongoing pregnancy rate per cycle in women with normal ovarian response was 70%. Considering that a non-inferiority threshold should retain 80% of the clinical effect of a control treatment, a minimal clinical difference of 14% (one-sided: α, 2.5%; β, 20%) and a total of 338 patients were needed. Anticipating a 10% drop-out rate, the total number of patients required was 372. This trial has been approved by the Institutional Ethical Committee of Beijing Chao-Yang hospital. All participants in the trial will provide written informed consent. The study will be conducted according to the principles outlined in the Declaration of Helsinki and its amendments. Results of this study will be disseminated in peer-reviewed scientific journals. ChiCTR1800018077.


Prediction of sperm retrieval in men with non-obstructive azoospermia using artificial neural networks: leptin is a good assistant diagnostic marker.
At present, non-invasive methods are not comprehensive enough to enable urologists to predict sperm retrieval results accurately in patients with non-obstructive azoospermia (NOA). Our aim was to improve the prediction accuracy of sperm retrieval by using leptin and artificial neural networks (ANNs). Data from May 2004 to July 2010 for 280 patients with NOA were reviewed and assigned into the training and testing set for ANNs. All patients underwent standard diagnostic infertility evaluation and testicular sperm extraction (TESE). Twelve factors were recorded as the input variables for ANNs: (1)testicular volume, (2)semen volume, seminal pH, seminal alpha-glucosidase and fructose, (3)serum hormones including FSH, LH, total testosterone (TT), prolactin, estradiol, (4)serum and seminal leptin. Three ANN models were constructed with the following input variables: ANN1-(1)(2)(3)(4), ANN2-(1)(2)(3) and ANN3-(1)(2)(4). The prediction accuracy for FSH, leptin and ANN models was compared by receiver operating characteristic (ROC) curve analysis. All ANN models were better than FSH. ANN1 had the largest area under the curve (AUC = 0.83) and demonstrated significant improvement compared with FSH (AUC = 0.63, P < 0.01) and leptin (AUC = 0.59, P< 0.01). ANNs improve the prediction accuracy of sperm retrieval. Although the leptin AUC is low, combined use of leptin and FSH can significantly improve the prediction accuracy for sperm recovery in NOA patients. Leptin may be a good assistant marker for diagnosing NOA. However, studies with larger numbers of patients are required to confirm the improved predictive performance of ANNs.


Protective Effects of the Segmental Renal Artery Clamping Technique on Ischemia-Reperfusion Injury in db/db Diabetic Mice.
Renal ischemia-reperfusion (I/R) injury is inevitable in partial nephrectomy and other kidney surgeries, with a higher incidence in patients with renal insufficiency. This study aimed to investigate the protective effects of precise segmental renal artery clamping (SRAC) against renal I/R injury in db/db diabetic mice, compared with conventional renal artery clamping (RAC). Grape seed extract, a powerful free radical scavenger, was administered to diabetic mice for 4 weeks before operation in subgroups (30 mg/kg/d). The unilateral renal pedicle was ligatured, and I/R injury to the contralateral kidney was induced (ischemia for 30 min followed by reperfusion for 24 h). Blood glucose value, creatinine, blood urea nitrogen, and urine microalbumin/urine creatinine ratio increased gradually and showed no preoperative statistical differences among six subgroups. These parameters were significantly lower in the SRAC than in the RAC group 24 h postoperatively. Moreover, the nonischemic area in the SRAC group expressed less KIM-1 and TNF-α mRNA and also revealed minor histopathological damage induced by I/R. These findings suggest that SRAC effectively reduces early renal injury induced by I/R and accelerates the recovery of renal function in diabetic mice. Thus, SRAC may be an ideal technique in partial nephrectomy, especially for patients with diabetic nephropathy and other renal insufficiencies.


Parental marital satisfaction in a family-based partial hospitalization program.
In family-based treatment, parents are initially put in charge of the weight restoration process, requiring parents to work closely together to help their child recover, possibly putting a strain on the marital relationship. The purpose of the current study was to examine marital satisfaction in families before and after treatment in a family-based program and to determine whether marital satisfaction is related to patient outcome. Parents of 53 adolescent patients participated in the study. Almost half of mothers and a third of fathers reported a decrease in marital satisfaction over the course of treatment. Descriptive analyses revealed that patients whose parents reported a decrease in marital satisfaction scored worse on a measure of eating disorder psychopathology. Further research is needed to determine the direction of causality.


A stable bis(methylene)-λ4-selane with a >C[double bond, length as m-dash]Se[double bond, length as m-dash]C< bond containing Se(iv).
Bis(methylene)-λ4-selane 1, which represents a new class of heteroallenes, was synthesized and isolated as a stable purple crystalline solid. X-ray crystallographic analysis revealed a bent allene-type structure with pseudo C2v symmetry and a bent C[double bond, length as m-dash]Se[double bond, length as m-dash]C moiety that contains a 3-center-4-electron π-bond. The NMR spectrum of 1 shows compelling evidence for a slow rotation of the Se[double bond, length as m-dash]C moieties at room temperature. The 77Se NMR spectrum of 1 contains one signal in the region characteristic for Se(iv) compounds.


Investment appraisal of technology innovations on dairy farm electricity consumption.
The aim of this study was to conduct an investment appraisal for milk-cooling, water-heating, and milk-harvesting technologies on a range of farm sizes in 2 different electricity-pricing environments. This was achieved by using a model for electricity consumption on dairy farms. The model simulated the effect of 6 technology investment scenarios on the electricity consumption and electricity costs of the 3 largest electricity-consuming systems within the dairy farm (i.e., milk-cooling, water-heating, and milking machine systems). The technology investment scenarios were direct expansion milk-cooling, ice bank milk-cooling, milk precooling, solar water-heating, and variable speed drive vacuum pump-milking systems. A dairy farm profitability calculator was combined with the electricity consumption model to assess the effect of each investment scenario on the total discounted net income over a 10-yr period subsequent to the investment taking place. Included in the calculation were the initial investments, which were depreciated to zero over the 10-yr period. The return on additional investment for 5 investment scenarios compared with a base scenario was computed as the investment appraisal metric. The results of this study showed that the highest return on investment figures were realized by using a direct expansion milk-cooling system with precooling of milk to 15°C with water before milk entry to the storage tank, heating water with an electrical water-heating system, and using standard vacuum pump control on the milking system. Return on investment figures did not exceed the suggested hurdle rate of 10% for any of the ice bank scenarios, making the ice bank system reliant on a grant aid framework to reduce the initial capital investment and improve the return on investment. The solar water-heating and variable speed drive vacuum pump scenarios failed to produce positive return on investment figures on any of the 3 farm sizes considered on either the day and night tariff or the flat tariff, even when the technology costs were reduced by 40% in a sensitivity analysis of technology costs.


Rabs, Rips, FIPs, and endocytic membrane traffic.
Rab GTPases, proteins belonging to the Ras-like small GTP-binding protein superfamily, have emerged as master regulators of cellular membrane transport. Rab11 GTPase, a member of the Rab protein family, plays a role in regulating various cellular functions, including plasma membrane recycling, phagocytosis, and cytokinesis. Rab11 acts by forming mutually exclusive complexes with Rab11-family binding proteins, known as FIPs. Rab11-FIP complexes serve a role of "targeting complexes" by recruiting various membrane traffic factors to cellular membranes. Recent studies have identified several Rab11-FIP complex-binding proteins that regulate distinct membrane traffic pathways.


"Mothers will be lucky if utmost receive a single scheduled postnatal home visit": An exploratory qualitative study, Northern Ethiopia.
Postnatal home visits (PNHVs) have been endorsed as strategy for delivery of postnatal care (PNC) to reduce newborn mortality and improve maternal outcomes. Despite the important role of the Health Extension Workers (HEWs) in improving the overall healthcare coverage, PNHV remains as a missed opportunity in rural Ethiopia. Thus, this study aimed to explore the barriers and facilitators of scheduled postnatal home visits in Northern Ethiopia. We conducted an exploratory qualitative study on a total of 16 in-depth interviews with HEWs and mothers who gave birth one year prior to the study. In addition, focus group discussions were conducted with HEWs and key informant interviews were conducted with women development group leaders, supervisors, and healthcare authorities from April to June 2019 in two rural districts of Northern Ethiopia. Discussions and interviews were audio recorded and transcribed verbatim in the local language (Tigrigna) and translated into English. The translated scripts were thematically coded using Atlas ti scientific software. Field notes were also taken during the discussion and while conducting the interviews. Health system factors, community context, and individual level factors were considered as the barriers and facilitators of scheduled PNHVs. Leadership, governance, management, support and supervision, referral linkages, overwhelming workload, capacity building, logistics and supplies are the major sub-themes identified as health system factors. Physical characteristics like geographical location and topography, distance, and coverage of the catchment; and community support and participation like support from women's development groups (WDGs), awareness of the community on the presence of the service and cultural and traditional beliefs were community contexts that affect PNHVs. Self-motivation to support and intrinsic job satisfaction were individual level factors that were considered as barriers and facilitators. The finding of this study suggested that the major barriers of postnatal home visits were poor attention of healthcare authorities of the government bodies, lack of effective supervision, poor functional linkages, inadequate logistics and supplies, unrealistic catchment area coverage, poor community participation and support, and lack of motivation of HEWs. Henceforth, to achieve the scheduled PNHV in rural Ethiopia, there should be strong political commitment and healthcare authorities should provide attention to postnatal care both at facility and home with a strong controlling system.


Biomarker panels for characterizing microbial community biofilm formation as composite molecular process.
Microbial consortia execute collaborative molecular processes with contributions from individual species, on such basis enabling optimized molecular function. Such collaboration and synergies benefit metabolic flux specifically in extreme environmental conditions as seen in acid mine drainage, with biofilms as relevant microenvironment. However, knowledge about community species composition is not sufficient for deducing presence and efficiency of composite molecular function. For this task molecular resolution of the consortium interactome is to be retrieved, with molecular biomarkers particularly suited for characterizing composite molecular processes involved in biofilm formation and maintenance. A microbial species set identified in 18 copper environmental sites provides a data matrix for deriving a cross-species molecular process model of biofilm formation composed of 191 protein coding genes contributed from 25 microbial species. Computing degree and stress centrality of biofilm molecular process nodes allows selection of network hubs and central connectors, with the top ranking molecular features proposed as biomarker candidates for characterizing biofilm homeostasis. Functional classes represented in the biomarker panel include quorum sensing, chemotaxis, motility and extracellular polysaccharide biosynthesis, complemented by chaperones. Abundance of biomarker candidates identified in experimental data sets monitoring different biofilm conditions provides evidence for the selected biomarkers as sensitive and specific molecular process proxies for capturing biofilm microenvironments. Topological criteria of process networks covering an aggregate function of interest support the selection of biomarker candidates independent of specific community species composition. Such panels promise efficient screening of environmental samples for presence of microbial community composite molecular function.


Tumour-Targeted and Redox-Responsive Mesoporous Silica Nanoparticles for Controlled Release of Doxorubicin and an siRNA Against Metastatic Breast Cancer.
Metastatic breast cancer seriously harms women's health and is currently the tumour type with the highest mortality rate in women. Recently, the combinatorial therapeutic approaches that integrate anti-cancer drugs and genetic agents is an attractive and promising strategy for the treatment of metastatic breast cancer. Moreover, such a combination strategy requires better drug carriers that can effectively deliver the cargo to the breast cancer cells and achieve controlled release in the cells to achieve better therapeutic effects. The tumour-targeted and redox-responsive mesoporous silica nanoparticles (MSNs) functionalised with DNA aptamers (AS1411) as a co-delivery system was developed and investigated for the potential against metastatic breast cancer. Doxorubicin (Dox) was loaded onto the MSNs, while AS1411 and a small interfering RNA (siTIE2) were employed as gatekeepers via attachment to the MSNs with redox-sensitive disulfide bonds. The controlled release of Dox and siTIE2 was associated with intracellular glutathione. AS1411 mediated the targeted delivery of Dox by increasing its cellular uptake in metastatic breast cancer, ultimately resulting in a lower IC50 in MDA-MB-231 cells (human breast cancer cell line with high metastatic potency), improved biodistribution in tumour-bearing mice, and enhanced in vivo anti-tumour effects. The in vitro cell migration/invasion assay and in vivo anti-metastatic study revealed synergism in the co-delivery system that suppresses cancer cell metastasis. The tumour-targeted and redox-responsive MSN prepared in this study are promising for the effective delivery and controlled release of Dox and siTIE2 for improved treatment of metastatic breast cancer.


[THE CORONARY SINUS REDUCER: A DEVICEBASED THERAPY FOR REFRACTORY ANGINA].
Alongside the remarkable progress in therapeutic strategies destined to patients with coronary artery disease (CAD), a growing number of patients who suffer from severe and diffuse CAD not amenable for revascularization, continue to experience severe debilitating angina despite optimal medical therapy. The Coronary Sinus (CS) Reducer has emerged as a novel therapeutic strategy for patients with refractory angina pectoris. The Reducer is a stainless steel, balloon-expandable, hourglass shaped device implanted percutaneously through the right internal jugular vein in the coronary sinus. After implantation it creates a controlled narrowing of the lumen, leading to an increase in coronary venues pressure, which improves blood flow to ischemic layers of the myocardium by redistribution of blood from the subepicardial myocardium to the more ischemic subendocardial layers of the myocardium, thus improving angina symptoms and quality of life.


Isoflurane causes anterograde but not retrograde amnesia for pavlovian fear conditioning.
Production of retrograde amnesia by anesthetics would indicate that these drugs can disrupt mechanisms that stabilize memory. Such disruption would allow suppression of memory of previous untoward events. The authors examined whether isoflurane provides retrograde amnesia for classic (Pavlovian) fear conditioning. Rats were trained to fear tone by applying three (three-trial) or one (one-trial) tone-shock pairs while breathing various constant concentrations of isoflurane. Immediately after training, isoflurane administration was either discontinued, maintained unchanged, or rapidly increased to 1.0 minimum alveolar concentration for 1 h longer. Groups of rats were similarly trained to fear context while breathing isoflurane by applying shocks (without tones) in a distinctive environment. The next day, memory for the conditioned stimuli was determined by presenting the tone or context (without shock) and measuring the proportion of time each rat froze (appeared immobile). For each conditioning procedure, the effects of the three posttraining isoflurane treatments were compared. Rapid increases in posttraining isoflurane administration did not suppress conditioned fear for any of the training procedures. In contrast, isoflurane administration during conditioning dose-dependently suppressed conditioning (P < 0.05). Training to tone was more resistant to the effects of isoflurane than training to context (P < 0.05), and the three-trial learning procedure was more was more resistant than the one-trial procedure (P < 0.05). Isoflurane provided intense dose-dependent anterograde but not retrograde amnesia for classic fear conditioning. Isoflurane appears to disrupt memory processes that occur at or within a few minutes of the conditioning procedure.


Subjective scoring of cough in children: parent-completed vs child-completed diary cards vs an objective method.
Cough is often used as an outcome measure, although the reporting of cough is unreliable. Using a 24 h ambulatory cough meter to measure cough frequency, the aim of this study was to compare: 1) the correlation of child-completed diary cards to the objective measurement, with that of parent-completed diary cards; and 2) the visual analogue scale (VAS) to the verbal category descriptive (VCD) score. The cough meter consisted of a previously validated Holter monitor and a cough processor. Eighty four children (39 with recurrent cough and 45 controls, aged 6-17 yrs) used a cough meter at least once. Thirty three subjects used the cough meter twice. Parents and children completed separate diary cards using the VAS and VCD scores. The strength of the relationship between the subjective scores and the objective recordings was analysed by spearman's rank correlation coefficient. For daytime cough, child-completed diary cards and the VCD correlated better to the objective measurement than parent-completed diary cards and the VAS, respectively. In subjects that used the cough meter twice, the difference between the cough frequency correlated to the difference in the subjective scores. The confidence intervals for the correlation coefficients were wide. The agreement between the objective and subjective presence of daytime cough was good but that for night-time cough was poor. We conclude that the severity of cough defined on diary cards may not represent cough frequency. Objective readings are first choice but currently not yet practical. The verbal category descriptive diary card completed by children and assisted by parents has the highest correlation to cough frequency measured objectively.


Safety of Hibitane. II. Human experience.
Chlorhexidine has been widely used in medical practice since its introduction on to the marked in the early 1950s. Primarily it has been used for topical antisepsis, e.g. pre-surgery skin preparation, burns prophylaxis, and prior to obstetrical/gynaecological procedures. This extensive experience has demonstrated the virtual absence of sensitization and a low irritancy potential for the compound. Only one significant adverse effect has been identified during medical use, namely, the production of sensorineural deafness after direct instillation into the middle ear, a property shared by several commonly used antiseptics. The encouraging reports of the use of chlorhexidine for plaque control prompted further safety investigations. It has been shown that absorption after oral ingestion is very low, and long-term oral use has not produced changes in haematological and biochemical parameters. Occasional oral intolerance of mouthrinse formulations has been reported, although no histological abnormalities were present in gingival biopsies taken after 18 months' daily use. Very occasionally, a reversible swelling of the parotid glands has been reported after use of chlorhexidine in mouthrinse formulations, but not after other methods of administration. Tooth discoloration, which shows wide inter-individual variation is seen after all dental formulations. This undesirable cosmetic effect would appear to represent the only significant argument against everyday prophylactic oral use of chlorhexidine.


Source monitoring reduces the suggestibility of preschool children.
The relation between source monitoring and suggestibility was examined among preschool children. Thirty-two 3- to 5-year-olds were simultaneously presented with a brief story in two different modalities, as a silent video vignette and a spoken narrative. Each modality presented unique information about the story, but the information in the two versions was mutually compatible. The children were then asked a series of questions, including questions about the source (modality) of story details, and leading questions about story details (to assess suggestibility). Performance on the source-monitoring questions was highly correlated with the ability to resist suggestion. In addition, children who were asked source-monitoring questions prior to leading questioning were less susceptible to suggestion than were those who were asked the leading questions first. This study provides evidence that source monitoring can play a causal role in reducing the suggestibility of preschool children.


Efficacy of Mobile Apps to Support the Care of Patients With Diabetes Mellitus: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.
Diabetes Mellitus (DM) is a chronic disease that is considered a global public health problem. Education and self-monitoring by diabetic patients help to optimize and make possible a satisfactory metabolic control enabling improved management and reduced morbidity and mortality. The global growth in the use of mobile phones makes them a powerful platform to help provide tailored health, delivered conveniently to patients through health apps. The aim of our study was to evaluate the efficacy of mobile apps through a systematic review and meta-analysis to assist DM patients in treatment. We conducted searches in the electronic databases MEDLINE (Pubmed), Cochrane Register of Controlled Trials (CENTRAL), and LILACS (Latin American and Caribbean Health Sciences Literature), including manual search in references of publications that included systematic reviews, specialized journals, and gray literature. We considered eligible randomized controlled trials (RCTs) conducted after 2008 with participants of all ages, patients with DM, and users of apps to help manage the disease. The meta-analysis of glycated hemoglobin (HbA1c) was performed in Review Manager software version 5.3. The literature search identified 1236 publications. Of these, 13 studies were included that evaluated 1263 patients. In 6 RCTs, there were a statistical significant reduction (P<.05) of HbA1c at the end of studies in the intervention group. The HbA1c data were evaluated by meta-analysis with the following results (mean difference, MD -0.44; CI: -0.59 to -0.29; P<.001; I²=32%).The evaluation favored the treatment in patients who used apps without significant heterogeneity. The use of apps by diabetic patients could help improve the control of HbA1c. In addition, the apps seem to strengthen the perception of self-care by contributing better information and health education to patients. Patients also become more self-confident to deal with their diabetes, mainly by reducing their fear of not knowing how to deal with potential hypoglycemic episodes that may occur.


Peripheral myelin maintenance is a dynamic process requiring constant Krox20 expression.
Onset of myelination in Schwann cells is governed by several transcription factors, including Krox20/Egr2, and mutations affecting Krox20 result in various human hereditary peripheral neuropathies, including congenital hypomyelinating neuropathy (CHN) and Charcot-Marie-Tooth disease (CMT). Similar molecular information is not available on the process of myelin maintenance. We have generated conditional Krox20 mutations in the mouse that allowed us to develop models for CHN and CMT. In the latter case, specific inactivation of Krox20 in adult Schwann cells results in severe demyelination, involving rapid Schwann cell dedifferentiation and increased proliferation, followed by an attempt to remyelinate and a block at the promyelinating stage. These data establish that Krox20 is not only required for the onset of myelination but that it is also crucial for the maintenance of the myelinating state. Furthermore, myelin maintenance appears as a very dynamic process in which Krox20 may constitute a molecular switch between Schwann cell myelination and demyelination programs.


Demographic patterns in Lyme borreliosis seasonality over 25 years.
Lyme borreliosis, the most common vector-borne disease in Europe and North America, is attracting growing concern due to its expanding geographic range. The growth in incidence and geographic spread is largely attributed to climate and land-use changes that support the tick vector and thereby increase disease risk. Despite a wide range of symptoms displayed by Lyme borreliosis patients, the demographic patterns in clinical manifestations and seasonal case timing have not been thoroughly investigated and may result from differences in exposure, immunity and pathogenesis. We analysed 25 years of surveillance data from Norway, supplemented by population demography data, using a Bayesian modelling framework. The analyses aimed to detect differences in case seasonality and clinical manifestations of Lyme borreliosis across age and sex differentiated patient groups. The results showed a bimodal pattern of incidence over age, where children (0-9 years) had the highest incidence, young adults (20-29 years) had low incidence and older adults had a second incidence peak in the ages 70-79 years. Youth (0-19 years) presented with a higher proportion of neuroborreliosis cases and a lower proportion of arthritic manifestations compared to adults (20+ years). Adult males had a higher overall incidence than adult females and a higher proportion of arthritis cases. The seasonal timing of Lyme borreliosis consistently occurred around 4.4 weeks earlier in youth compared to adults, regardless of clinical manifestation. All demographic groups exhibited a shift towards an earlier seasonal timing over the 25-year study period, which appeared unrelated to changes in population demographics. However, the disproportionate incidence of Lyme borreliosis in seniors requires increased public awareness and knowledge about this high-risk group as the population continues to age concurrently with disease emergence. Our findings highlight the importance of considering patient demographics when analysing the emergence and seasonal patterns of vector-borne diseases using long-term surveillance data.


Comparison of Nanoscale Focused Ion Beam and Electrochemical Lithiation in β-Sn Microspheres.
The development of Li focused ion beams (Li-FIB) enables controlled Li ion insertion into materials with nanoscale resolution. We take the first step toward establishing the relevance of the Li-FIB for studies of ion dynamics in electrochemically active materials by comparing FIB lithiation with conventional electrochemical lithiation of isolated β-Sn microspheres. Samples are characterized by cross-sectioning with Ga FIB and imaging via electron microscopy. The Li-FIB and electrochemical lithiated Sn exhibit similarities that suggest that the Li-FIB can be a powerful tool for exploring dynamical Li ion-material interactions at the nanoscale in a range of battery materials.


Oral vaccination based on DNA-chitosan nanoparticles against Schistosoma mansoni infection.
The development of a vaccine would be essential for the control of schistosomiasis, which is recognized as the most important human helminth infection in terms of morbidity and mortality. A new approach of oral vaccination with DNA-chitosan nanoparticles appears interesting because of their great stability and the ease of target accessibility, besides chitosan immunostimulatory properties. Here we described that chitosan nanoparticles loaded with plasmid DNA encoding Rho1-GTPase protein of Schistosoma mansoni, prepared at different molar ratios of primary amines to DNA phosphate anion (N/P), were able to complex electrostatically with DNA and condense it into positively charged nanostructures. Nanoparticles were able to maintain zeta potential and size characteristics in media that simulate gastric (SGF) and intestinal fluids (SIF). Further in vivo studies showed that oral immunization was not able to induce high levels of specific antibodies but induced high levels of the modulatory cytokine IL-10. This resulted in a significative reduce of liver pathology, although it could not protect mice of infection challenge with S. mansoni worms. Mice immunized only with chitosan nanoparticles presented 47% of protection against parasite infection, suggesting an important role of chitosan in inducing a protective immune response against schistosomiasis, which will be more explored in further studies.


Persistence of socioeconomic inequalities in the knowledge of cardiovascular risk factors five years after coronary angiography.
Cardiovascular diseases are important causes of death, morbidity, and years of potential life lost in most developed countries. The purpose of this study was to assess trends in knowledge of cardiovascular risk factors among patients five years after coronary angiography and to investigate the impact of educational level on knowledge level. The study included 1289 of 4391 patients admitted for cardiac events in 2008/2009 at the National Institute for Cardiac Surgery and Interventional Cardiology, Luxembourg. A follow-up study was conducted by post five years later (2013/2014). Data were obtained from 1837 of the contacted patients (with 548 reported deaths) (response rate=42%). Logistic regression models were used to evaluate the association between educational level and knowledge of cardiovascular risk factors. Educational level was used as a surrogate for socioeconomic status. In total, 39.9% of patients could list at least three risk factors in 2013/2014, a much higher percentage than the 8.5% observed during the initial survey. In both sexes, knowledge of cardiovascular risk factors increased between 2008/2009 and 2013/2014. Patients with higher educational levels were more likely (odds ratio=2.33, 95% confidence interval: 1.63-3.34) to cite at least three risk factors than patients with lower education levels. Knowledge level was associated with educational level, and improved for all educational groups five years after coronary angiography. Educational differences in knowledge persisted, but the gaps decreased. Improving knowledge of cardiovascular risk factors among patients with cardiovascular disease will help increase awareness and promote lifestyle changes.


Fluorescent Immunohistochemistry: An Important Tool to Reveal Proteins From Tissues in Ancient Mummified Remains.
During the restoration of the Saint Antonio Abate Cathedral in Castelsardo, Sardinia, Italy, numerous human remains were found in a crypt. The burial site contained upwards of 120 individuals organized in successive layers from the bottom of the crypt; of these, 18 partially mummified individuals have been identified, including the last 2 individuals buried in the crypt. In the present study, we focused on these 2 individuals by initially adopting a morphologic and anthropological approach. The anthropological analysis of the remains showed that the 2 bodies were partially mummified and that most of the organs were not available; for this reason, the sex was determined by secondary sexual characteristics of the skulls and the long bones. The aim of this research was to describe the general state of the mummified bodies and tissues by morphologic and ultrastructural analysis using light and electron microscopy techniques. To ensure the preservation of specific tissue proteins, immunohistochemical fluorescence analysis was used. Limited information is available regarding the preservation of mummified tissues. Thus, this study demonstrated the presence of muscle and skin tissue markers in a good state of preservation, even though the tissues had undergone a slow mummification process. Our results demonstrate that several types of tissues and cell proteins may survive over a prolonged period and that these materials survive the postmortem processes.


Palliative surgery for brain metastases of malignant melanoma.
Forty patients with melanoma brain metastases were treated by surgery. Single brain metastases were found in 32 cases and multiple in eight. The most frequent tumor location was the frontal and frontoparietal lobes. Neurological improvement was observed in 25 patients and surgical mortality rate was less than 5%. The median survival time for all patients was 8 months. When patients with multiple cerebral metastases were excluded the median survival time was 13 months. The 3- and 5-year survival was 25% and 15%, respectively. Seventeen patients with extracerebral metastases received treatment and were without known extracerebral tumor at the time of brain metastases diagnosis. These patients had a median survival time that did not significantly differ from those without occurrence of extracerebral metastases. Quality of life as judged by Karnofsky index was improved after surgery and maintained on an acceptable level for the remaining time of survival.


Comprehensive Drug Screening of Whole Blood by LC-HRMS-MS in a Forensic Laboratory.
As the number of prescriptions, over-the-counter medications and drugs of abuse continue to increase, forensic laboratories are faced with the challenge of developing more comprehensive screening methods in order to detect them in whole blood samples. Another challenge faced by forensic laboratories is detecting and identifying novel synthetic compounds as they emerge and change. Traditional drug screening methods include enzyme immunoassay (EIA) and either gas or liquid chromatography paired with mass spectrometry (GC-MS or LC-MS-MS, respectively). While these methods are good, they have their disadvantages. For example, EIA requires special reagents for each drug class, GC-MS requires extensive sample preparation, and LC-MS-MS only detects drugs on a known inclusion lists of compounds of interest. Described here is the development of a robust and comprehensive screening method for drugs in whole blood samples that eliminates the aforementioned disadvantages of the traditional methods. Using a Q Exactive Focus™ liquid chromatography-high-resolution accurate mass spectrometer (LC-HRMS-MS), a method was developed that is capable of detecting ~200 drugs at a concentration of 2 μg/L for most analytes. This method also employs a more automated data processing feature which reduces processing time. Finally, it has the added benefit of retroactive data analysis, which allows it to be used for unknown drug analysis as well. Used as an initial screening method, the comprehensive drug screen using LC-HRMS-MS has the potential to take on two of the most important challenges faced by forensic laboratories today.


Frequency-resolving spatiotemporal wave-front sensor.
We report on a high-resolution wave-front sensor that measures the complete spatial profile of any frequency component of a laser field containing multiple frequencies. This probe technique was developed to address the necessity of measuring the spatial overlap of the carrier field with each sideband component of the field exiting the output port of a gravitational-wave interferometer. We present the results of an experimental test of the probe, where we were able to construct the spatial profile of a single radio-frequency sideband at the level of -50 dBc.


Intergenerational cultural conflict, mental health, and educational outcomes among Asian and Latino/a Americans: Qualitative and meta-analytic review.
Among immigrant Asian and Latino groups, the contrast between collectivism in traditional heritage and individualism in the mainstream American cultures presents unique challenges for their family relationships. This systematic review was designed to answer 3 fundamental questions: to what extent do(es) (a) acculturation mismatch (AM) correlate with intergenerational cultural conflict (ICC); (b) ICC correlate with offspring's mental health and educational outcomes; and (c) demographic and study characteristics moderate these relationships. Sixty-one research reports were reviewed, with 68 independent study samples (N = 14,453; 41 and 27 Asian and Latino/a samples, respectively) subjected to 3 meta-analyses. AM positively correlated with ICC (r = .23), which in turn negatively correlated with offspring mental health (r = -.20) and educational outcomes (r = -.09). Findings provided support for acculturation gap-distress theory. While these effect size estimates were small, participant and methodological variables affected their magnitude. Contrary to findings on intergenerational conflict within mainstream non-immigrant families, the relationships among AM, ICC, and mental health were larger in young adult than adolescent groups within immigrant families. ICC significantly correlated with internalizing problems and adaptive functioning, but not externalizing problems. AM was more closely related to ICC among women and second-generation immigrant offspring. AM and ICC were more problematic among offspring who were low-risk and lived in less ethnically disperse regions, particularly when studied in cross-sectional studies. Effect sizes also differed significantly across measurement tools for the key constructs. Limitations to generalizability (few studies on educational outcomes, relative under-representation of Latino/a to Asian American samples), and implications for intervention and future research are discussed.


Interfacial engineered iron oxide nanoring for T2-weighted magnetic resonance imaging-guided magnetothermal-chemotherapy.
Due to no penetration depth limitation, low cost, and easy control, magnetic nanoparticles mediated magnetic hyperthermia therapy (MHT) has shown great potential in experimental and clinal treatments of various diseases. However, the low heating conversion efficiencies and short circulation times are major drawback for most existing magnetic-thermal materials. Additionally, single MHT treatment always leads to resistance and recurrence. Herein, a highly efficient magnetic-thermal conversion, ferrimagnetic vortex nanoring Fe3O4 coated with hyaluronic acid (HA) nanoparticles (Fe3O4@HA, FVNH NPs) was firstly constructed. Additionally, the doxorubicin (DOX) was successfully enclosed inside the FVNH and released remotely for synergetic magnetic-thermal/chemo cancer therapy. Due to the ferrimagnetic vortex-domain state, the ring shape Fe3O4 displays a high specific absorption rate (SAR) under an external alternating magnetic field (AMF). Additionally, antitumor drug (DOX) can be encapsulated inside the single large hole of FVNH by the hyaluronic acid (HA) shell and quickly released in response the tumor acidic microenvironments and AMF. What's more, the non-loaded FVNH NPs show good biocompatibility but high cytotoxicity after loading DOX under AMF. Furthermore, the synthesized FVNH can efficiently reduce the transverse relaxation time and enhance negative magnetic resonance imaging (MRI). The impressive in vivo systemic therapeutic efficacy of FVNH was also proved in this work. Taken together, the results of this study demonstrate that the synthesized FVNH NPs offer the promise of serving as multifunctional theranostic nanoplatforms for medical imaging-guided tumor therapies.


Treatment strategy based on the natural course of the disease for patients with spontaneous isolated abdominal aortic dissection.
This study aimed to determine the natural history of spontaneous isolated abdominal aortic dissection (SIAAD) and to establish an optimal management strategy for patients with SIAAD. We searched the database of thoracoabdominal computed tomography (CT) performed at a single institution from January 2003 to July 2016 using the keywords "aortic dissection" and "dissection AND aorta." Once a diagnosis of SIAAD was made, we investigated the initial clinical and morphologic features and aorta-related events for all patients and morphologic changes of the aortic dissection (AD) during the follow-up period for the patients who underwent follow-up CT scans. We compared characteristics of the patients, frequencies of clinical events (aortic rupture, intervention, death), and morphologic changes (false lumen enlargement, progression of AD, remodeling of AD, and involvement of iliac or visceral artery) during the follow-up period according to the location of AD (infrarenal vs suprarenal), symptom status (symptomatic vs asymptomatic), and gender. There were 210 (10.7%) patients (median age, 69.4 years [interquartile range, 61.3-74.7]; male, 73.3%) who were diagnosed with SIAAD among 1958 patients with AD. SIAAD was most frequently located at the infrarenal aorta (86.2%), extended to the iliac (12.4%) or visceral artery (2.9%), and was symptomatic in 13.3% of patients. During the study period, aortic rupture developed in two patients (0.9%), aortic intervention was required in five (2.4%), and aorta-related deaths were identified in three (1.4%). Among 138 (65.7%) patients who underwent follow-up CT scans, 81.9% showed no morphologic change or remodeling during the follow-up period (median, 25 months; range, 1-158 months; interquartile range, 12.3-49.1 months). In the meantime, false lumen enlargement and longitudinal progression of AD developed in 8.7% and 6.5% of patients, respectively. However, newly developed visceral artery extension was not found in any of the patents. When characteristics of the patients and frequencies of clinical events or late morphologic changes of AD were compared on the basis of the location of AD, symptom status, and gender, female gender, presence of symptoms, and suprarenal SIAAD were more frequently associated with aorta-related mortality. False lumen enlargement was more frequent in the suprarenal AD group than in the infrarenal AD group. Based on our observation, the majority of symptomatic and asymptomatic SIAAD patients can be managed conservatively unless they present with aortic rupture, concomitant large aortic aneurysm, or underlying connective tissue disease. However, a more proactive management strategy may be required for female, symptomatic patients or those with suprarenal SIAAD.


Precipitation of oppositely charged polyelectrolytes in salt solutions.
We study phase separation in symmetric solutions of weakly charged flexible chains of opposite sign. Precipitation is caused by effective attractions due to charge fluctuations and by short-range attractions between monomers. The contribution from charge fluctuations is computed within the random phase approximation (RPA), which takes into account the connectivity of charges in the polyions. The impenetrability of the ions is accounted for by using a modified Coulomb potential in the RPA. In good solvent conditions the precipitate monotonically swells and eventually dissolves upon addition of salt. However, near the theta-solvent condition, but still in the good solvent, the precipitate can be stable at any salt concentration. Moreover, the density of the precipitate after initial decrease can increase with addition of salt. This effect is a result of redistribution of salt between the precipitate and the supernatant, which is due to an interplay of electrostatic and hardcore interactions. For not too weakly charged polyions the precipitate properties become strongly dependent on temperature even in good solvent conditions.


Hematologic effects of xylazine when used for restraint of Bactrian camels.
Xylazine was found to be a safe and reliable drug for chemical restraint of Bactrian camels (Camelus bactrianus). Dosages of 0.27 to 0.51 mg/kg of body weight provided adequate sedation for the performance of various procedures (e.g., tuberculin testing, lymph node biopsy, and electroejaculation). Hematologic and serum biochemical values for camels restrained manually were compared with those for camels restrained with xylazine. Xylazine-treated camels had significantly lower values for RBC, hemoglobin, and packed cell volume, and significantly higher blood glucose concentrations. Venous blood gas analyses did not reveal any major acid-base disturbances resulting from the use of xylazine. Rapid arousal from the sedative effects of xylazine occurred after the intravenous administration of doxapram hydrochloride in dosages of 0.05 to 0.13 mg/kg of body weight.


[The status of organ transplantation in France and Europe in November 1987].
The number of organ transplantations in France is constantly and impressively increasing, but this progress remains insufficient. Compared with other European countries, France ranks first as regards the number of heart transplantations performed in 1984 but is lagging behind as regards kidney transplantations. Since potential donors, the France-Transplant organization and qualified teams of operators are available, further advances are to be expected from a reinforcement of the facilities available to France-Transplants.


Bioimage informatics: a new area of engineering biology.
In recent years, the deluge of complicated molecular and cellular microscopic images creates compelling challenges for the image computing community. There has been an increasing focus on developing novel image processing, data mining, database and visualization techniques to extract, compare, search and manage the biological knowledge in these data-intensive problems. This emerging new area of bioinformatics can be called 'bioimage informatics'. This article reviews the advances of this field from several aspects, including applications, key techniques, available tools and resources. Application examples such as high-throughput/high-content phenotyping and atlas building for model organisms demonstrate the importance of bioimage informatics. The essential techniques to the success of these applications, such as bioimage feature identification, segmentation and tracking, registration, annotation, mining, image data management and visualization, are further summarized, along with a brief overview of the available bioimage databases, analysis tools and other resources.


Pretibial epidermolysis bullosa: report of two families and review of the literature.
Two pretibial epidermolysis bullosa families, with a total of four cases directly examined and another eight cases known from references, were studied. Both families showed an autosomal dominant inheritance. In one family there was a case of "bullous epidermolysis albo-papular of Pasini". All cases were compared with those of the literature.


The Negative Effects of Volatile Sulphur Compounds.
Oral malodor has been studied extensively in humans but not necessarily to the same degree in our veterinary patients where malodor constitutes a significant problem. Breath malodor may originate from the mouth, or from an extra oral source, originating from other organ systems such as gastrointestinal, respiratory, or even systemic disease. Oral malodor is a result of microbial metabolism of exogenous and endogenous proteinaceous substrates leading to the production of compounds such as indole, skatole, tyramine, cadaverine, puterescine, mercaptans, and sulphides. Volatile sulphur compounds have been shown to be the main cause of oral malodor. Although most clients perceive oral malodor to be primarily a cosmetic problem, there is an increasing volume of evidence in human dental literature demonstrating that volatile sulphur compounds produced by bacteria, even at low concentrations, are toxic to tissues and play a role in the pathogenesis of periodontitis. This article reviews the current available literature in human dentistry looking at these negative effects. No veterinary studies have been conducted looking at the negative effects of volatile sulphur compounds specifically, but as this article highlights, we should be aware of the potential negative effects of volatile sulphur compounds and consider this an area of future research.


Composite 5A zeolite with ultrathin porous TiO2 coating for selective gas adsorption.
A composite zeolite adsorbent was prepared by conformally depositing an ultrathin porous TiO2 coating on the external surface of the 5A zeolite by molecular layer deposition (MLD) and subsequent calcination. The composite adsorbent showed significantly improved ideal adsorption selectivity for CO2-CH4, CO2-N2 and propylene-propane.


Proteomic analysis of the oxidative stress response in Kluyveromyces lactis and effect of glutathione reductase depletion.
Yeasts are unicellular eukaryotes that provide useful models for studying the oxidative stress (OS) response. Most investigations to date have been performed on the fermentative Saccharomyces cerevisiae. The respiratory Kluyveromyces lactis is emerging as an alternative model. Our previous studies showed that glutathione reductase (Glr1) is an interesting point of difference in the OS response between the two yeasts. In the present study, using extensive proteomic analyses, the response to H(2)O(2) and its relationship to Glr1 were investigated in wild-type and glr1-deletion mutant K. lactis strains. We identified 46 proteins that showed modified expression after H(2)O(2) addition and 42 for which the change was Glr1-dependent. As expected, these proteins include a variety of antioxidant enzymes, chaperones, and oxidoreductases related to defense against OS and damage repair. They also include a number of proteins necessary for energy production and carbohydrate and amino acid metabolism. H(2)O(2) addition causes down-regulation of enzymes from the glycolytic pathway and Krebs cycle in wild-type K. lactis, whereas glr1-deletion prevents this effect and actually causes up-regulation of the glycolytic, Krebs cycle, and oxidative pentose phosphate pathways. To our knowledge, this is the first global proteomic analysis performed on K. lactis.


Physical activity as a moderator for obstructive sleep apnoea and cardiometabolic risk in the EPISONO study.
Obstructive sleep apnoea (OSA) is positively associated with cardiometabolic diseases; however, high levels of physical activity could decrease the incidence of OSA and associated comorbidities.In this study we aimed to examine the incidence of OSA in relation to physical activity, and its role as a protective factor in individuals with OSA on the incidence of cardiometabolic diseases, in an 8-9-year follow-up study. We analysed data of 658 volunteers from the São Paulo Epidemiologic Sleep Study (EPISONO), a cohort study of individuals aged 20-80 years, collected through polysomnography, the International Physical Activity Questionnaire and an assessment of cardiometabolic profile.Active subjects had a lower risk of developing OSA compared with nonactive subjects (relative risk 0.877, 95% CI 0.296-0.855) and there was a reduced risk of developing type 2 diabetes mellitus in active/apnoeic subjects (relative risk 0.493, 95% CI 0.252-0.961) compared with nonactive subjects. Metabolic equivalent was negatively associated to cardiometabolic markers, such as C-reactive protein (exp(B)=0.720; p=0.001), interleukin-6 (exp(B)=0.991; p=0.03), insulin (exp(B)=0.982; p=0.03), triglycerides (exp(B)=0.997; p<0.001), homeostasis model assessment for insulin resistance (exp(B)≤0.946; p<0.024), quantitative insulin sensitivity check index (exp(B)=992.4; p<0.001) and mean arterial pressure (exp(B)=0.987; p=0.001).Physical activity was a protective factor against type 2 diabetes mellitus in apnoeic individuals; moreover, being active reduced the risk of developing OSA and was associated with a better cardiometabolic profile.


Identification of immune-related biomarkers co-occurring in acute ischemic stroke and acute myocardial infarction.
Acute ischemic stroke (AIS) and acute myocardial infarction (AMI) share several features on multiple levels. These two events may occur in conjunction or in rapid succession, and the occurrence of one event may increase the risk of the other. Owing to their similar pathophysiologies, we aimed to identify immune-related biomarkers common to AIS and AMI as potential therapeutic targets. We identified differentially expressed genes (DEGs) between the AIS and control groups, as well as AMI and control groups using microarray data (GSE16561 and GSE123342). A weighted gene co-expression network analysis (WGCNA) approach was used to identify hub genes associated with AIS and/or AMI progression. The intersection of the four gene sets identified key genes, which were subjected to functional enrichment and protein-protein interaction (PPI) network analyses. We confirmed the expression levels of hub genes using two sets of gene expression profiles (GSE58294 and GSE66360), and the ability of the genes to distinguish patients with AIS and/or AMI from control patients was assessed by calculating the receiver operating characteristic values. Finally, the investigation of transcription factor (TF)-, miRNA-, and drug-gene interactions led to the discovery of therapeutic candidates. We identified 477 and 440 DEGs between the AIS and control groups and between the AMI and control groups, respectively. Using WGCNA, 2,776 and 2,811 genes in the key modules were identified for AIS and AMI, respectively. Sixty key genes were obtained from the intersection of the four gene sets, which were used to identify the 10 hub genes with the highest connection scores through PPI network analysis. Functional enrichment analysis revealed that the key genes were primarily involved in immunity-related processes. Finally, the upregulation of five hub genes was confirmed using two other datasets, and immune infiltration analysis revealed their correlation with certain immune cells. Regulatory network analyses indicated that GATA2 and hsa-mir-27a-3p might be important regulators of these genes. Using comprehensive bioinformatics analyses, we identified five immune-related biomarkers that significantly contributed to the pathophysiological mechanisms of both AIS and AMI. These biomarkers can be used to monitor and prevent AIS after AMI, or vice versa.


[Trends of Concept Development in Nursing Published in Korean Journals].
The purpose of this study was to identify trends of nursing concept development in Korean journal papers to improve accurate understanding of nursing concepts. A systematic review of 216 concept development articles published from 1970 to 2018 that met the inclusion criteria was conducted using Research Information Sharing Service (RISS) databases. The most common method of concept development was Walker and Avant's concept analysis method, identified in 139 (64.3%) of the 216 studies, followed by 48 examples of hybrid models (22.2%) and 15 examples of evolutionary methods (6.9%). Chinn and Kramer's method, Norris's clarification, Wilson's method, and others were also used. The concepts of "spirituality" and "fatigue" were most frequently analyzed. Among the 139 studies that used Walker and Avant's concept analysis method, 127 studies (91.4%) applied all the recommended steps; the others applied the recommended steps partially, omitting description of model cases/additional cases, antecedents/consequences, and empirical indicators. Among the studies using the hybrid model, among two (5.7%) did not describe attributes, three (8.5%) did not provide definitions, and 16 (45.7%) did not present empirical indicators in the final stage. Among concept development studies published in Korean journals, Walker and Avant's concept analysis method is most commonly used. In case of most studies using Walker and Avant's method a suitable concept analysis process is applied, but in case of other studies using the other concept development method, a suitable concept analysis process is not applied. Therefore, a suitable concept analysis process must be applied for concept development in nursing research.


Perceived discrimination and depressive symptoms among immigrant-origin adolescents.
Although discrimination has been found to contribute to psychological distress among immigrant populations, there are few studies that have examined the relationship between racial and ethnic discrimination in the school setting among foreign-born immigrant and U.S.-born immigrant-origin adolescents. This study examined the relationship between perceived discrimination by adults and peers in the school setting and depressive symptoms in a sample (N = 95) of racial minority immigrant-origin adolescents (13 to 19 years of age) attending an urban high school. We examined the relation between perceived discrimination and depressive symptomology across gender and nativity status (foreign born vs. U.S. born), and the potential moderating role of ethnic identity and social support. Consistent with previous research, girls reported higher levels of depressive symptomology than boys, although the relationship between perceived discrimination and depressive symptoms was significant for both boys and girls. Perceived discrimination by adults and by peers at school was positively related to depressive symptoms for U.S.-born adolescents. For U.S.-born adolescents, ethnic identity mitigated the negative effects of perceived adult discrimination on depressive symptoms. However, ethnic identity did not moderate the relationship between perceived peer discrimination and depressive symptoms. Social support did not moderate the relationship between adult and peer discrimination and depressive symptoms for either foreign-born or U.S.-born adolescents. The findings support previous research concerning the immigrant paradox and highlight the importance of context in the relationship between perceived discrimination and mental health. Implications for future research and intervention are discussed.


An endothelin receptor antagonist TAK-044 ameliorates carbon tetrachloride-induced acute liver injury and portal hypertension in rats.
Hepatic levels of a powerful vasoconstrictor endothelin-1 (ET-1) and its receptors increase in human and carbon tetrachloride (CCl4)-induced liver cirrhosis. The aim of this study was to determine whether antagonism of hepatic ET-1 receptors ameliorates CCl4-induced hepatic injury and portal hypertension in rats. Acute liver injury was induced by a single intraperitoneal injection of CCl4 (0.3 ml/kg), whereas cirrhosis and portal hypertension were induced by CCl4 treatment (0.15 ml/kg twice a week) for 8 weeks. Hepatic morphology, ET-1 and its receptors, and portal venous pressures were determined. Increases in ET-1 and its receptors occurred within 24 h of CCl4 administration, and progressively thereafter during the development of cirrhosis. The acute CCl4-induced hepatic injury was characterized by significant increases in portal pressure (from 8.7+/-1.8 to 17.6+/-3.3 mmHg; p<0.01) and serum levels of liver enzymes, as well as massive hepatocellular necrosis (62+/-8%). Intravenous administration of an ET-1 receptor antagonist TAK-044 reduced portal pressure to 13.6+/-2.8 mmHg (p<0.05), and ameliorated hepatocellular necrosis by about 35% (p<0.001). TAK-044 treatment also produced significant reduction in serum levels of liver enzymes. In cirrhotic rats, portal venous infusion of TAK-044 reduced portal hypertension by about 40% (p<0.05). In conclusion, these results indicate involvement of ET-1 in acute liver injury as well as portal hypertension associated with hepatic cirrhosis, and a potential for ET-1 receptor antagonists in the treatment of these pathologic conditions.


Dihydrofolate Reductase Genetic Polymorphisms Affect Methotrexate Dose Requirements in Pediatric Patients With Acute Lymphoblastic Leukemia on Maintenance Therapy.
We have aimed to determine the effect of polymorphisms in regulatory regions of the DHFR gene in relation to methotrexate (MTX) dose adjustments and drug-induced toxicity in children on maintenance therapy for acute lymphoblastic leukemia (ALL). In total, 41 children diagnosed with ALL were screened for 3 tag-single nucleotide polymorphisms in the DHFR promoter (C-1610G, C-680G/T, A-317G) and an intronic 19-bp insertion/deletion. Genotypes were analyzed in relation to dose requirements and toxicity. The percentage of MTX dose administered (with respect to protocol-recommended values) was affected by DHFR polymorphisms. Carriers of the -680AA genotype displayed a median percentage of 44.08 (interquartile range=34.69), compared with 77.98 (interquartile range=33.90) for CC and CA carriers (P=0.01). The number of counts within white blood cell therapeutic range (2.0 to 3.0×10/L) was higher for -680AA carriers than for CC/CA carriers (P=0.003). With regard to toxicity, carriers of the -680AA genotype displayed more treatment interruptions than CC/CG carriers (P=0.03), as well as more episodes of severe neutropenia (P=0.04) and higher number of blood counts with elevated levels (>400 mg/dL) of lactate dehidrogenase (P=0.04). Overall, our findings suggest that the identification of DHFR polymorphisms in the promoter region of the gene may be helpful in tailoring MTX doses for ALL pediatric patients on maintenance therapy.


Concurrent Afatinib and Whole-Brain Radiotherapy in Exon 19-del-EGFR Mutant Lung Adenocarcinoma: A Case Report and Mini Review of the Literature.
Leptomeningeal metastases (LM) are found in approximately 3.8% of non-small cell lung cancer cases with an increased incidence in adenocarcinoma, and approximately one-third of patients will present with concomitant brain metastases. We report the case of a 50-year-old male patient with stage IV exon 19-del-EGFR mutant lung adenocarcinoma who progressed on second-generation TKI therapy with manifestation of symptomatic simultaneous diffuse brain and LM. Whole-brain radiotherapy with concurrent afatinib resulted in an almost complete regression of neurological symptoms as well as good, durable radiological response. Furthermore, treatment was well tolerated with no relevant adverse effects.


Metallacryptate single-molecule magnets: effect of lower molecular symmetry on blocking temperature.
The structural characterization of complexes [Mn(II)4Mn(III)22(pdol)12(OCH3)12(O)16(N3)6] (1) and [Mn(II)4Mn(III)22(pdol)12(OCH3)12(O)16(OH)2(H3O)(OCH3)3].ClO4.5CH3OH (2), where pdol(2-) is di-2-pyridyl methanediol, reveals that each has a metallacryptand shell that encapsulates a manganese oxide core. Variable-temperature direct current magnetic susceptibility measurements on 2 indicate a paramagnetic ground state that results from an overall antiferromagnetic interaction in the cluster, with chiT values decreasing from 300 K (51.2 cm3 K mol(-1)) to 2 K (19.8 cm3 K mol(-1)). Variable-temperature alternating current magnetic susceptibility measurements imply that both 1 and 2 behave as single-molecule magnets. Fitting the frequency-dependent out-of-phase magnetic susceptibility to the Arrhenius equation yields an effective energy barrier, Ueff, to magnetization relaxation of 16.5 +/- 0.7 K (11.5 +/- 0.5 cm(-1)) for 1 and 36.2 +/- 2.0 K (25.1 +/- 1.4 cm(-1)) for 2. The larger value for 2 is in agreement with the lower molecular symmetry, larger magnetoanisotropy, and higher ground spin state of 2 compared to those of 1. This observation suggests a new strategy for increasing the blocking temperatures in high-nuclearity manganese clusters.


BLX-1002, a novel thiazolidinedione with no PPAR affinity, stimulates AMP-activated protein kinase activity, raises cytosolic Ca2+, and enhances glucose-stimulated insulin secretion in a PI3K-dependent manner.
BLX-1002 is a novel small thiazolidinedione with no apparent affinity to peroxisome proliferator-activated receptors (PPAR) that has been shown to reduce glycemia in type 2 diabetes without adipogenic effects. Its precise mechanisms of action, however, remain elusive, and no studies have been done with respect to possible effects of BLX-1002 on pancreatic beta-cells. We have investigated the influence of the drug on beta-cell function in mouse islets in vitro. BLX-1002 enhanced insulin secretion stimulated by high, but not low or intermediate, glucose concentrations. BLX-1002 also augmented cytoplasmic free Ca2+ concentration ([Ca2+](i)) at high glucose, an effect that was abolished by pretreatment with the Ca2+-ATPase inhibitor thapsigargin. In contrast, BLX-1002 did not interfere with voltage-gated Ca2+ channel or ATP-sensitive K+ channel activities. In addition, cellular NAD(P)H stimulated by glucose was not affected by the drug. The stimulatory effect of BLX-1002 on insulin secretion at high glucose was completely abolished by treatment with the phosphatidylinositol 3-kinase (PI3K) inhibitors wortmannin or LY-294002. Stimulation of the beta-cells with BLX-1002 also induced activation of AMP-activated protein kinase (AMPK) at high glucose. Our study suggests that BLX-1002 potentiates insulin secretion only at high glucose in beta-cells in a PI3K-dependent manner. This effect of BLX-1002 is associated with an increased [Ca2+](i) mediated through Ca2+ mobilization, and an enhanced activation of AMPK. The glucose-sensitive stimulatory impact of BLX-1002 on beta-cell function may translate into substantial clinical benefits of the drug in the management of type 2 diabetes, by avoidance of hypoglycemia.


Endogenous pneumoconiosis: Analytical scanning electron microscopic analysis of a case.
Pneumoconiosis is often considered a disease of the lung initiated by exposure to dust or other airborne particles, resulting in injury to the lungs. The term "endogenous pneumoconiosis" has been used in the literature to describe the deposition of compounds on the elastic fibers of the lung, usually in the setting of cardiac failure. In the case we present here, the patient aspirated a foreign body resulting in damage to the lung tissue and subsequent deposition of endogenous compounds on the elastic fibers of the pulmonary parenchyma and vasculature. We determined the composition of this mineral and mapped the distribution of elements using a combination of backscattered electron microscopy and energy dispersive spectrometry.


Combined use of fluorescent peanut agglutinin lectin and Hoechst 33258 to monitor the acrosomal status and vitality of human spermatozoa.
Fluorescein-conjugated peanut agglutinin (PNA) lectin-labelling is an established procedure for assessing the status of the human sperm acrosome. However, unlike the triple-stain technique, PNA-labelling does not provide a simultaneous assessment of cellular vitality. We have therefore evaluated the use of the fluorescent dye Hoechst 33258 (H33258) as a vital stain for use in combination with PNA-labelling. Human sperm populations were stained for 1 min with 1 microgram/ml H33258 in culture medium and then washed through 2.0 (w/v) polyvinylpyrollidone columns and air-dried onto microscope slides. H33258 was found to provide vitality assessments comparable to those obtained using the standard eosin-exclusion method. However, best results were obtained with an ethanol fixation step between air-drying and PNA-labelling. This vitality assessment was found to be more reliable than that provided by Trypan blue staining under conditions equivalent to the triple-stain technique. There was no alteration of PNA-labelling due to the H33258 although ethanol fixation actually provided more uniform PNA-labelling than previously obtained without ethanol fixation. Consequently, we have stopped using the triple-stain technique for assessing human sperm acrosome reactions and now use the H33258/PNA procedure routinely.


Effects of hypothalamic stimulation on gluconeogenesis and glycolysis in rat liver.
Hypothalamic effects on gluconeogenesis and glycosis in rat liver were studied byelectrical stimulations of the ventromedial hypothalamic nucleus (LH) without disturbing the animal's behavior. Stimulation of VMH caused a increase in the activity of phosphoenolpyruvate carboxykinase (PEPCK), a key gluconeogenic enzyme, and marked suppression of pyruvate kinase (PK), a key glycolytic enzyme, of the liver. Stimulationof LH, on the other hand, resulted in a decrease in PEPCK activity but did not alterPK activity. The maximal responses of these enzymes to hypothalamic stimulations were obtained after intermittent stimulations for 4 h. Differential estimations of thetwo isozymes of liver PK (types L and M) were made using antibody against typeM PK. The level of type M enzyme was not altered significantly on stimulation of either VMH or LH. The content of type L enzyme greatly decreased on stimulation of VMH but was not affected by stimulation of LH. The reciprical influences of VMH and LH on hepatic carbohydrate metabolism and their relation to neural-hormonal responses are discussed.


Cloning and characterization of archaeal type I signal peptidase from Methanococcus voltae.
Archaeal protein trafficking is a poorly characterized process. While putative type I signal peptidase genes have been identified in sequenced genomes for many archaea, no biochemical data have been presented to confirm that the gene product possesses signal peptidase activity. In this study, the putative type I signal peptidase gene in Methanococcus voltae was cloned and overexpressed in Escherichia coli, the membranes of which were used as the enzyme source in an in vitro peptidase assay. A truncated, His-tagged form of the M. voltae S-layer protein was generated for use as the substrate to monitor the signal peptidase activity. With M. voltae membranes as the enzyme source, signal peptidase activity in vitro was optimal between 30 and 40 degrees C; it was dependent on a low concentration of KCl or NaCl but was effective over a broad concentration range up to 1 M. Processing of the M. voltae S-layer protein at the predicted cleavage site (confirmed by N-terminal sequencing) was demonstrated with the overexpressed archaeal gene product. Although E. coli signal peptidase was able to correctly process the signal peptide during overexpression of the M. voltae S-layer protein in vivo, the contribution of the E. coli signal peptidase to cleavage of the substrate in the in vitro assay was minimal since E. coli membranes alone did not show significant activity towards the S-layer substrate in in vitro assays. In addition, when the peptidase assays were performed in 1 M NaCl (a previously reported inhibitory condition for E. coli signal peptidase I), efficient processing of the substrate was observed only when the E. coli membranes contained overexpressed M. voltae signal peptidase. This is the first proof of expressed type I signal peptidase activity from a specific archaeal gene product.


Mutation of a nucleosome compaction region disrupts Polycomb-mediated axial patterning.
Nucleosomes play important structural and regulatory roles by tightly wrapping the DNA that constitutes the metazoan genome. The Polycomb group (PcG) proteins modulate nucleosomes to maintain repression of key developmental genes, including Hox genes whose temporal and spatial expression is tightly regulated to guide patterning of the anterior-posterior body axis. CBX2, a component of the mammalian Polycomb repressive complex 1 (PRC1), contains a compaction region that has the biochemically defined activity of bridging adjacent nucleosomes. Here, we demonstrate that a functional compaction region is necessary for proper body patterning, because mutating this region leads to homeotic transformations similar to those observed with PcG loss-of-function mutations. We propose that CBX2-driven nucleosome compaction is a key mechanism by which PcG proteins maintain gene silencing during mouse development.


Confounding and heterogeneity in genetic association studies with admixed populations.
Association studies among admixed populations pose many challenges including confounding of genetic effects due to population substructure and heterogeneity due to different patterns of linkage disequilibrium (LD). We use simulations to investigate controlling for confounding by indicators of global ancestry and the impact of including a covariate for local ancestry. In addition, we investigate the use of an interaction term between a single-nucleotide polymorphism (SNP) and local ancestry to capture heterogeneity in SNP effects. Although adjustment for global ancestry can control for confounding, additional adjustment for local ancestry may increase power when the induced admixture LD is in the opposite direction as the LD in the ancestral population. However, if the induced LD is in the same direction, there is the potential for reduced power because of overadjustment. Furthermore, the inclusion of a SNP by local ancestry interaction term can increase power when there is substantial differential LD between ancestry populations. We examine these approaches in genome-wide data using the University of Southern California's Children's Health Study investigating asthma risk. The analysis highlights rs10519951 (P = 8.5 × 10(-7)), a SNP lacking any evidence of association from a conventional analysis (P = 0.5).


Particulate matter emissions, and metals and toxic elements in airborne particulates emitted from biomass combustion: The importance of biomass type and combustion conditions.
The aim of this study was to investigate the impact of biomass combustion with respect to burning conditions and fuel types on particulate matter emissions (PM10) and their metals as well as toxic elements content. For this purpose, different lab scale burning conditions were tested (20 and 13% O2 in the exhaust gas which simulate an incomplete and complete combustion respectively). Furthermore, two pellet stoves (8.5 and 10 kW) and one open fireplace were also tested. In all cases, 8 fuel types of biomass produced in Greece were used. Average PM10 emissions ranged at laboratory-scale combustions from about 65 to 170 mg/m3 with flow oxygen at 13% in the exhaust gas and from 85 to 220 mg/m3 at 20% O2. At pellet stoves the emissions were found lower (35 -85 mg/m3) than the open fireplace (105-195 mg/m3). The maximum permitted particle emission limit is 150 mg/m3. Metals on the PM10 filters were determined by several spectrometric techniques after appropriate digestion or acid leaching of the filters, and the results obtained by these two methods were compared. The concentration of PM10 as well as the total concentration of the metals on the filters after the digestion procedure appeared higher at laboratory-scale combustions with flow oxygen at 20% in the exhaust gas and even higher at fireplace in comparison to laboratory-scale combustions with 13% O2 and pellet stoves. Modern combustion appliances and appropriate types of biomass emit lower PM10 emissions and lower concentration of metals than the traditional devices where incomplete combustion conditions are observed. Finally, a comparison with other studies was conducted resulting in similar results.


Limits to using HPA axis activity as an indication of animal welfare.
HPA axis activity is often measured by corticosteroid release as a means to evaluate stress and well-being in animals. While the analysis of corticosteroid levels can provide useful information in some circumstances, a variety of methodological and technical problems make them difficult to accurately interpret. Furthermore, there is considerable evidence that is inconsistent with the widespread notion that high levels of stress result in a large amount of corticosteroid release, and in some cases the converse is true. This review highlights the strengths and weaknesses of the techniques used to measure corticosteroids, describes a number of studies that failed to find a positive correlation between stress and corticosteroid levels, and delineates ancillary behavioral and cognitive tests that provide insight into an animal's well-being. We conclude by emphasizing that the most holistic account of animal welfare is provided by utilizing a combination of physiological and psychological methods.


CD4⁺ and CD8⁺ regulatory T cells (Tregs) are elevated and display an active phenotype in patients with chronic HCV mono-infection and HIV/HCV co-infection.
The aim of this study was to examine regulatory T cells (Tregs) in peripheral blood and liver tissue in patients with chronic hepatitis C virus (HCV) mono-infection and in patients with HIV/HCV co-infection. In a cross-sectional study were included 51 patients with chronic HCV infection, 24 patients with HIV/HCV co-infection and 24 healthy individuals. CD4⁺ and CD8⁺ Tregs were determined using flow cytometry. Fibrosis was examined by transient elastography. Inflammation, fibrosis and Tregs were determined in liver biopsies from 12 patients. Increased frequency of CD4⁺ and CD8⁺ Tregs was found in HIV/HCV co-infected patients [median: 6.4% (IQR: 5.7-6.9) and 1.0% (0.7-1.2), respectively] compared to HCV mono-infected patients [5.6% (4.2-6.3), P = 0.01 and 0.5% (0.3-0.7), P < 0.001, respectively]. Furthermore, HCV mono-infected patients had increased frequencies of Tregs compared with healthy controls (P < 0.05). However, no associations between the frequency of Tregs and fibrosis were found. Furthermore, characterization of CD4⁺ Tregs using CD45RA demonstrated a higher frequency of activated Tregs in both HCV mono-infected and HIV/HCV co-infected patients compared with healthy controls. Finally, number of intrahepatic Tregs was associated with both peripheral CD8⁺ Tregs and intrahepatic inflammation. In conclusion, HCV mono-infected patients and particularly HIV/HCV co-infected patients have increased the frequency of CD4⁺ and CD8⁺ Tregs compared with healthy controls. Furthermore, CD4⁺ Tregs in infected patients displayed an active phenotype. Tregs were not associated with fibrosis, but a positive correlation between intrahepatic Tregs and inflammation was found. Taken together, these results suggest a role for Tregs in the pathogenesis of chronic HCV infection.


Sarcopaenia and rheumatoid arthritis.
In October 2016 a new independent disease called sarcopaenia (according to ICD-10 classification) appeared. According to the recommendation of the European Working Group on Sarcopenia in Older People (EWGSOP), sarcopaenia is defined as abnormally low muscle mass plus low skeletal muscle strength or low physical performance. Sarcopaenia, as a primary disease, is mainly observed in older people, but it can also appear in younger adults in the course of many clinical chronic conditions. One of the most frequent chronic diseases associated with chronic inflammation and functional limitation of skeletal system is rheumatoid arthritis. In the present article, current knowledge on the epidemiology of sarcopaenia and its association with rheumatoid arthritis is presented.


Selective Oxidation of 2-Methylnaphthalene to 2-Methyl-l-naphthol by Rhodococcus sp. M192.
About 6000 isolates of microorganisms assimilating methylketones (C3-C6) were tested for their selective oxidation of 2-methylnaphthalene to 2-methyl-1-naphthol. Strain M192 was the highest 2-methyl-1-naphthol producer and was classified as the genus Rhodococcus. The optimal conditions for the site-specific oxidation were studied using resting Rhodococcus sp. M192. The 2-methyl-1-naphthol productivity was specifically increased using methylethylketone as a carbon source, 1-propanol as a solvent to dissolve the substrate, and ethylxanthate or diethyldithiocarbamate as an inhibitor of 2-naphthoic acid (side-product) production. In the presence of these compounds, 2-methylnaphthalene was specifically oxidized at the 1-position without the conversion to 2-naphthoic acid. The productivity of 2-methyl-1-naphthol was about 90 μM from 1 mM 2-methylnaphthalene.


Organoids as a new model system to study neural tube defects.
The neural tube is the first critically important structure that develops in the embryo. It serves as the primordium of the central nervous system; therefore, the proper formation of the neural tube is essential to the developing organism. Neural tube defects (NTDs) are severe congenital defects caused by failed neural tube closure during early embryogenesis. The pathogenesis of NTDs is complicated and still not fully understood even after decades of research. While it is an ethically impossible proposition to investigate the in vivo formation process of the neural tube in human embryos, a newly developed technology involving the creation of neural tube organoids serves as an excellent model system with which to study human neural tube formation and the occurrence of NTDs. Herein we reviewed the recent literature on the process of neural tube formation, the progress of NTDs investigations, and particularly the exciting potential to use neural tube organoids to model the cellular and molecular mechanisms underlying the etiology of NTDs.


[Cardiac magnetic resonance imaging with contrast enhancement in treatment of atrial fibrillation].
Catheter ablation is presently the main method for interventional treatment of atrial fibrillation (AF). Despite improvements of the method and accumulation of personnel's experience, incidence of recurrent AF following catheter interventions remains high. This review addresses a possibility of using contrast-enhanced cardiac magnetic resonance imaging to increase the effectiveness of interventional treatment of arrhythmia.


Production of ethanol by Saccharomyces cerevisiae under high-pressure conditions.
A research project was initiated to examine the possibility of using supercritical carbon dioxide for in situ recovery of ethanol during its production by yeast Saccharomyces cerevisiae. As a preliminary step, it was necessary to study the behavior of ethanol production under high-pressure conditions, up to 7 MPa (1000 psi). The results show that pressure has a significant inhibiting effect on the production of ethanol. There is a significant decrease in the initial rate of production as well as in the final ethanol concentration as pressure is increased. This decrease is more significant when carbon dioxide is used to pressurize the fermentor. The pressure affects the ability of the cells to produce ethanol in a reversible way. When the fermentor is returned to atmospheric conditions, the reaction resumes its normal fermentation rate.


Robot-assisted radical cystectomy with intracorporeal urinary diversion versus open radical cystectomy (iROC): protocol for a randomised controlled trial with internal feasibility study.
Bladder cancer (BC) is a common malignancy and one of the most expensive to manage. Radical cystectomy (RC) with pelvic lymphadenectomy is a gold standard treatment for high-risk BC. Reductions in morbidity and mortality from RC may be achieved through robot-assisted RC (RARC). Prospective comparisons between open RC (ORC) and RARC have been limited by sample size, use of extracorporeal reconstruction and use of outcomes important for ORC. Conversely, while RARC is gaining in popularity, there is little evidence to suggest it is superior to ORC. We are undertaking a prospective randomised controlled trial (RCT) to compare RARC with intracorporeal reconstruction (iRARC) and ORC using multimodal outcomes to explore qualitative and quantitative recovery after surgery. METHODS AND ANALYSIS: iROC is a multicentre prospective RCT in English National Health Service (NHS) cancer centres. We will randomise 320 patients undergoing RC to either iRARC or ORC. Treatment allocation will occur after trial entry and consent. The primary outcome is days alive and out of hospital within the first 90 days from surgery. Secondary outcomes will measure functional recovery (activity trackers, chair-to-stand tests and health related quality of life (HRQOL) questionnaires), morbidity (complications and readmissions), cost-effectiveness (using EuroQol-5 Domain-5 levels (EQ-5D-5L) and unit costs) and surgeon fatigue. Patients will be analysed according to intention to treat. The primary outcome will be transformed and analysed using regression. All statistical assumptions will be investigated. Secondary outcomes will be analysed using appropriate regression methods. An internal feasibility study of the first 30 patients will evaluate recruitment rates, acceptance of randomised treatment choice, compliance outcome collection and to revise our sample size. The study has ethical approval (REC reference 16/NE/0418). Findings will be made available to patients, clinicians, funders and the NHS through peer-reviewed publications, social media and patient support groups. ISRCTN13680280 and NCT03049410.


IL-12-mediated transcriptional regulation of matrix metalloproteinases.
Matrix metalloproteinases (MMPs) are extracellular matrix (ECM) remodelling enzymes involved in developmental processes, tissue remodelling and repair, inflammatory and immune diseases and cancer. In a recent issue of Bioscience Reports (vol. 37, issue 6, BSR20170973), Liu and colleagues investigated the expression of MMPs such as MMP-1 (interstitial collagenase), MMP-3 (stromelysin 1) and MMP-13 (collagenase 3) in human periodontal ligament fibroblasts (hPDLFs) regulated by interleukin-12 (IL-12), a cytokine implicated in inflammatory and immune responses. They showed that IL-12 activates canonical nuclear factor-κB (NF-κB) signalling leading to increased expression of MMP-1, MMP-3 and MMP-13, and to a smaller reduction in the expression of MMP-2 (gelatinase A) and MMP-9 (gelatinase B) at both mRNA and protein levels, with corresponding changes in the secreted levels of these ECM-remodelling and immune regulatory metalloproteinases. While canonical NF-κB signalling regulates these MMPs, it also interacts with additional factors to determine whether some of these MMPs are induced or downregulated, in response to IL-12. Here, we comment on the possible mechanisms of IL-12-mediated transcriptional regulation of MMPs.