Scout triggered multiple reaction monitoring mass spectrometry for the rapid transfer of large multiplexed targeted methods in metabolomics.
The field of metabolomics based on mass spectrometry has grown considerably in recent years due to the need to detect and, above all, quantify a very large number of metabolites, simultaneously. Up to now, targeted multiplexed analysis on complex samples by Liquid Chromatography coupled with tandem Mass Spectrometry (LC-MS/MS) has relied almost exclusively on compound detection based on absolute retention times, as in the Scheduled-MRM (sMRM) approach. Those methods turn out to be poorly transferable from one instrument to another and result in a time-consuming and tedious method development involving a significant number of critical parameters that need specific re-optimisation. To address this challenge, we introduce a novel acquisition mode called scout-triggered MRM (stMRM). In stMRM, a marker transition is used to trigger MS analysis for a group of dependent target analytes. These marker transitions are strategically distributed throughout the chromatographic run, and the dependent analytes are associated based on their retention times. The result is a targeted assay that remains robust even in the presence of retention time shifts. A 3 to 5-fold increase in the number of detected transitions associated to plasma metabolites was obtained when transferring from a direct application of a published sMRM to a stMRM method. This significant improvement highlights the universal applicability of the stMRM method, as it can be implemented on any LC system without the need for extensive method development. We subsequently illustrate the robustness of stMRM in modified chromatographic elution conditions. Despite a large change in metabolite's selectivity, the multiplexed assay successfully recovered 70% of the monitored transitions when consequently modifying the gradient method. These findings demonstrate the versatility and adaptability of stMRM, opening new avenues for the development of highly multiplexed LC-MS/MS methods in metabolomics. These methods are characterized by their analytical transparency and straightforward implementation using existing literature data.


Randomized clinical trials with levosimendan.
There are important unmet needs in the treatment of acute heart failure syndromes (AHFS). The unique dual mechanism of action of levosimendan suggests that this new agent may help fill some of these unmet needs. A review of randomized, controlled clinical trials with levosimendan demonstrated that it is well tolerated, and its use results in significantly reduced pulmonary capillary wedge pressure (PCWP) and increased cardiac output. Effects of levosimendan on PCWP and cardiac output are more pronounced than those observed with dobutamine. Levosimendan treatment is also associated with significantly improved clinical symptoms. Moreover, data from 3 trials indicate that levosimendan treatment was associated with improved 6-month survival compared with dobutamine treatment or placebo. Emerging data suggest that levosimendan is beneficial for patients with acute myocardial ischemia. Thus, early clinical indicators suggest that levosimendan may help prevent myocardial injury during hospitalization for AHFS and may be well suited for first-line therapy for AHFS.


Acquired Gerbode Defects Associated with Infective Endocarditis.
Infective endocarditis is associated with a high risk of mortality and complications. A very rare complication is the development of an acquired left ventricle to right atrium (Gerbode) defect secondary to bacterial invasion of the membranous septum. We report two complex cases of infective endocarditis complicated by acquired Gerbode defects, outlining the role of two-dimensional (2D) and three-dimensional (3D) transoesophageal imaging.


Propensity Score Matched Outcome Analysis of Lobar Ablation Versus Completion Thyroidectomy in Low-Risk Differentiated Thyroid Cancer Patients: Median Follow-Up of 11 Years.
Background: Radioactive iodine lobar ablation (RAILA) of the contralateral thyroid lobe has been suggested as an alternative to completion thyroidectomy in patients of differentiated thyroid cancer (DTC) who underwent initial hemithyroidectomy. However, data on long-term outcomes are scarce. In this study, we intended to describe the long-term outcomes of RAILA versus completion thyroidectomy in a cohort of low-risk DTC patients. Methods: Data of patients with low-risk DTC who underwent initial hemithyroidectomy, and were subsequently treated with either completion thyroidectomy or RAILA between 1996 and 2015, were collected and analyzed. The treatment outcomes included ablation rate, recurrence rate, recurrence-free survival (RFS), and adverse events, and were validated by propensity score matching analysis. Results: Of the 1243 patients (median age: 34 years, range: 5-78) with low-risk DTC, 514 patients underwent upfront RAILA while 729 patients underwent completion thyroidectomy followed by remnant ablation. The ablation rate following the first radioactive iodine (131I) cycle was 75.3% [95% confidence interval (CI) 71.3-78.9] in the RAILA group versus 84.1% [CI 81.2-86.6] in the completion thyroidectomy group (p < 0.001). Over median follow-up of 11.4 years (interquartile range: 8.3-15.8), the recurrence rates between the two groups were not significantly different (1.6% [CI 0.7-3.2] vs. 1.0% [CI 0.4-2.1], respectively, p = 0.343). The product limit estimate of RFS at 10 years was 98.6% [CI 97.6-99.6%] in the RAILA group versus 99.1% [CI 98.3-99.9%] in the completion thyroidectomy group (p = 0.391). The outcomes in 497 matched pairs generated through propensity score analysis were similar. None of the patients in the RAILA group experienced permanent hypocalcemia or recurrent laryngeal nerve palsy, while the corresponding rates in the completion thyroidectomy group were 10/729 (1.4%) (p = 0.006) and 5/729 (0.7%) (p = 0.080), respectively. Radiation thyroiditis was seen in 25/514 (4.9%) patients in the RAILA group versus 3/729 (0.4%) in the completion thyroidectomy group (p < 0.001). Conclusions: Upfront RAILA is seen to be a noninvasive, safe, and effective alternative to surgical completion thyroidectomy in low-risk DTC patients in the absence of macroscopic malignancy in the remaining thyroid lobe.


Multidrug resistance crisis during COVID-19 pandemic: Role of anti-microbial peptides as next-generation therapeutics.
The decreasing effectiveness of conventional drugs due to multidrug-resistance is a major challenge for the scientific community, necessitating development of novel antimicrobial agents. In the present era of coronavirus 2 (COVID-19) pandemic, patients are being widely exposed to antimicrobial drugs and hence the problem of multidrug-resistance shall be aggravated in the days to come. Consequently, revisiting the phenomena of multidrug resistance leading to formulation of effective antimicrobial agents is the need of the hour. As a result, this review sheds light on the looming crisis of multidrug resistance in wake of the COVID-19 pandemic. It highlights the problem, significance and approaches for tackling microbial resistance with special emphasis on anti-microbial peptides as next-generation therapeutics against multidrug resistance associated diseases. Antimicrobial peptides exhibit exceptional mechanism of action enabling rapid killing of microbes at low concentration, antibiofilm activity, immunomodulatory properties along with a low tendency for resistance development providing them an edge over conventional antibiotics. The review is unique as it discusses the mode of action, pharmacodynamic properties and application of antimicrobial peptides in areas ranging from therapeutics to agriculture.


Divalent ions as mediators of carbonylation in cardiac myosin binding protein C.
The dosing and efficacy of chemotherapeutic drugs can be limited by toxicity caused by off-pathway reactions. One hypothesis for how such toxicity arises is via metal-catalyzed oxidative damage of cardiac myosin binding protein C (cMyBP-C) found in cardiac tissue. Previous research indicates that metal ion mediated reactive oxygen species induce high levels of protein carbonylation, changing the structure and function of this protein. In this work, we use long timescale all-atom molecular dynamics simulations to investigate the ion environment surrounding the C0 and C1 subunits of cMyBP-C responsible for actin binding. We show that divalent cations are co-localized with protein carbonylation-prone amino acid residues and that carbonylation of these residues can lead to site-specific interruption to the actin-cMyBP-C binding.


Gene expression profiling of human colon xenograft tumors following treatment with SU11248, a multitargeted tyrosine kinase inhibitor.
Biomarkers that indicate biological activity and/or efficacy are a potentially useful tool in the development of molecularly targeted therapeutics. It is useful, though challenging, to identify biomarkers during preclinical development in order to impact decision-making during early clinical development. SU11248 is an oral, selective multitargeted tyrosine kinase inhibitor currently in Phase II oncology clinical trials. It exhibits direct antitumor and antiangiogenic activity via inhibition of the receptor tyrosine kinases PDGFR, VEGFR, KIT and FLT3. To identify clinically translatable biomarkers of SU11248 activity, expression profiling was performed on Colo205 human xenograft tumors following treatment with SU11248. Over 100 transcripts changed in abundance in SU11248 as compared to vehicle-treated tumors. Nine candidate transcripts, chosen based on putative function, were also analysed and validated by TaqMan. One such potential biomarker, cadherin-11, was further evaluated at the protein level and was found to have increased expression in xenograft tumors after SU11248 treatment. Interestingly, cadherin-11 expression was also detected via immunohistochemical analysis of archived solid tumors, indicating the technical feasibility of translating this putative biomarker to clinical studies. Importantly, SU11248 treatment also resulted in increased expression of cadherin-11 protein in human tumor biopsies in three out of seven patients examined and confirms the feasibility of using transcriptional profiling of preclinical models to identify clinically translatable biomarkers.


Hydrolysis of nucleoside phosphates: IV. The metal ion-nucleic base interaction in the Cu2+-promoted dephosphorylation of the 5'-di- and 5'-triphosphates of cytidine, inosine and guanosine, and their protection toward hydrolysis by coordination to Cu(2,2'-bipyridyl)2+.
The dephosphorylation of CTP, GTP, ITP, ATP, CDP, GDP, IDP and ADP was characterized by measuring the first-order rate constant (50 degrees; I = 0.1, NaClO4) in dependence on pH (2 to 10). Except with CTP and CDP, the reactions are significantly accelerated by Cu2+ and pass through pH optima. By computing the pH dependence of the distribution of the several species present in the nucleotide (NP) systems, it is shown that the most reactive species is Cu(NP). Cu(NP-H), where N(1) is deprotonated, is somewhat less reactive. In both types of complexes, a metal ion-nucleic base interaction, which is responsible for the increased reactivity, occurs, i.e., macrochelates involving the phosphate chains and the base moieties are formed. In accord herewith, CTP and CDP are rather stable as the coordination tendency of the cytosine moiety is small. Furthermore, in the ternary complexes Cu(2,2'-bipyridyl)(NP) and Cu-(2,2'-bipyridyl)(NP-H), where the formation of a macrochelate is inhibited, the nucleotides are protected. The structure-reactivity relationship is also evident with Cu(ITP)2- and Cu(IDP)- which exist only in part as macrochelates; hence, they are less reactive than for example Cu(ATP)2- or Cu(ADP)-. With the aid of the initial rate, vo = d[PO4(3-)]/dt, the rate laws of the ascending side of the pH optima were determined: vo = k[Cu(NP)]/[H+]. A reaction mechanism that includes an intermolecular attack of OH- at the terminal phosphate group is proposed. The descending side of the pH optimum is attributed to the formation of CU(NP)(OH) or Cu(NP-H)(OH), where the Cu2+-base interaction is insignificant. However, these hydroxy complexes are still somewhat faster dephosphorylated than the free nucleotides. This is attributed to an intramolecular attack of the bound OH- at the terminal phosphate group.


Surgical Treatment of Upper Extremity Pain.
If the patient with hand pain remains without significant relief and without recovery of function after appropriate pharmaceutical and physical modality treatments, it is appropriate to consider a surgical approach to the pain. Categories of pain amenable to a surgical approach are pain caused by nerve compression, pain caused by a neuroma, and joint pain of neural origin. Compressed nerve should be decompressed and depending on the intraoperative findings a neurolysis also should be performed. Painful neuroma must be resected to stop the pain generator. For a painful joint, the biomechanics of that joint must first be stable before denervation.


New taxa of the subtribe Lebinthina (Orthoptera: Gryllidae: Eneopterinae: Lebinthini) from New Guinea and nearby islands.
Four new species from three genera of Lebinthina crickets are described here. These include one species of Gnominthus: Gnominthus milneus sp. nov. from Papua New Guinea; two new species of Macrobinthus: Macrobinthus kei sp. nov. and Macrobinthus mamai sp. nov. from Maluka (Indonesia) and Papua New Guinea, respectively; and one species of Microbinthus: Microbinthus elegans sp. nov. from Papua New Guinea.


Human milk fat content: within-feed variation.
Changes in milk fat content during individual breast-feeds were studied using a modification of the interrupted feed technique (Woolridge et al. (1982): Early Human Dev., 6, 265-272). Each feed was interrupted once, for weighing the baby and obtaining a small milk sample; weighing and milk sampling were also done at the start and end of the feed from that breast. Mathematical analysis of 52 feeds from the first breast offered at a feeding episode, and 39 second-breast feeds, showed a similar pattern on both types. The fat content rose in line with (Vi/Ve)1.35 (V being the volume consumed since the feed from that breast began). However, first-breast feeds showed sharp initial and final rises in fat content. The authors propose a mechanism to explain the difference in pattern between first- and second-breast feeds.


Identification of rearranged sequences of HPV16 DNA in precancerous and cervical cancer cases.
Integration of HPV16 DNA into the host chromosome is considered to be a crucial step towards genomic instability and cervical cancer development. Aim of the present study was to investigate the presence of HPV16 rearranged intra-viral sequences in HPV16-positive normal, precancerous and cervical cancer samples using the method of Restriction Site-PCR (RS-PCR). Sequence analysis of HPV16 integrants revealed for the first time in clinical samples two distinct rearranged intra-viral sequences, concerning the conjunction of E2 and L1 genes and the conjunction of E1 and L1 genes with inverted orientation. Furthermore mapping analysis of the E1 and E2 genes in cervical samples with rearranged intra-viral sequences of HPV16 genome was conducted in order to determine the integrity of viral genes. The identification of intra-viral rearrangements provides valuable information regarding the HPV16 DNA integration, and may be a significant biomarker for the presence of chromosomal instability and DNA damages in clinical samples.


Thirty-day mortality after ischemic stroke and intracranial hemorrhage in patients with atrial fibrillation on and off anticoagulants.
Prescribing warfarin for atrial fibrillation depends in large part on the expected reduction in ischemic stroke risk versus the expected increased risk of intracranial hemorrhage (ICH). However, the anticoagulation decision also depends on the relative severity of such events. We assessed the impact of anticoagulation on 30-day mortality from ischemic stroke versus ICH in a large community-based cohort of patients with atrial fibrillation. We followed 13,559 patients with atrial fibrillation enrolled in an integrated healthcare delivery system for a median 6 years. Incident ischemic strokes and ICHs were identified from computerized databases and validated through medical record review. The association of warfarin and international normalized ratio at presentation with 30-day mortality was modeled using multivariable logistic regression adjusting for clinical factors. We identified 1025 incident ischemic strokes and 299 ICHs during follow-up. Compared with no antithrombotic therapy, warfarin was associated with reduced Rankin score and lower 30-day mortality from ischemic stroke (adjusted OR, 0.64; 95% CI, 0.45-0.91) but a higher mortality from ICH (OR, 1.62; 95% CI, 0.88-2.98). Therapeutic international normalized ratios (2-3) were associated with an especially low ischemic stroke mortality (OR, 0.38; 95% CI, 0.20-0.70), whereas international normalized ratios>3 increased the odds of dying of ICH by 2.66-fold (95% CI, 1.21-5.86). Warfarin reduces 30-day mortality from ischemic stroke but increases ICH-related mortality. Both effects on event severity as well as on event rates need to be incorporated into rational decision-making about anticoagulants for atrial fibrillation.


Biology and Assembly of the Bacterial Envelope.
All free-living bacterial cells are delimited and protected by an envelope of high complexity. This physiological barrier is essential for bacterial survival and assures multiple functions. The molecular assembly of the different envelope components into a functional structure represents a tremendous biological challenge and is of high interest for fundamental sciences. The study of bacterial envelope assembly has also been fostered by the need for novel classes of antibacterial agents to fight the problematic of bacterial resistance to antibiotics. This chapter focuses on the two most intensively studied classes of bacterial envelopes that belong to the phyla Firmicutes and Proteobacteria. The envelope of Firmicutes typically has one membrane and is defined as being monoderm whereas the envelope of Proteobacteria contains two distinct membranes and is referred to as being diderm. In this chapter, we will first discuss the multiple roles of the bacterial envelope and clarify the nomenclature used to describe the different types of envelopes. We will then define the architecture and composition of the envelopes of Firmicutes and Proteobacteria while outlining their similarities and differences. We will further cover the extensive progress made in the field of bacterial envelope assembly over the last decades, using Bacillus subtilis and Escherichia coli as model systems for the study of the monoderm and diderm bacterial envelopes, respectively. We will detail our current understanding of how molecular machines assure the secretion, insertion and folding of the envelope proteins as well as the assembly of the glycosidic components of the envelope. Finally, we will highlight the topics that are still under investigation, and that will surely lead to important discoveries in the near future.


A predictive model for dysphagia following IMRT for head and neck cancer: introduction of the EMLasso technique.
Design a model for prediction of acute dysphagia following intensity-modulated radiotherapy (IMRT) for head and neck cancer. Illustrate the use of the EMLasso technique for model selection. Radiation-induced dysphagia was scored using CTCAE v.3.0 in 189 head and neck cancer patients. Clinical data (gender, age, nicotine and alcohol use, diabetes, tumor location), treatment parameters (chemotherapy, surgery involving the primary tumor, lymph node dissection, overall treatment time), dosimetric parameters (doses delivered to pharyngeal constrictor (PC) muscles and esophagus) and 19 genetic polymorphisms were used in model building. The predicting model was achieved by EMLasso, i.e. an EM algorithm to account for missing values, applied to penalized logistic regression, which allows for variable selection by tuning the penalization parameter through crossvalidation on AUC, thus avoiding overfitting. Fifty-three patients (28%) developed acute ≥ grade 3 dysphagia. The final model has an AUC of 0.71 and contains concurrent chemotherapy, D2 to the superior PC and the rs3213245 (XRCC1) polymorphism. The model's false negative rate and false positive rate in the optimal operation point on the ROC curve are 21% and 49%, respectively. This study demonstrated the utility of the EMLasso technique for model selection in predictive radiogenetics.


Robustness of the partial nitrification-anammox system exposing to triclosan wastewater: Stress relieved by extracellular polymeric substances and resistance genes.
The partial nitrification-anammox (PN/A) process is a promising method for the treatment of municipal wastewater. It is necessary to clarify the responses of PN/A system to antimicrobial agent triclosan (TCS) widely existed in the influent of wastewater treatment plants. In this study, it was found that PN/A system was robust to cope with 0.5 mg/L TCS. Specifically, the control reactor reached 80% total nitrogen removal efficiency (TNRE) on day 107, while the reactor feeding with 0.5 mg/L TCS reached the same TNRE on day 84. The results of the activity test, high-throughput sequencing and DNA-based stable isotope probing showed that 0.5 mg/L TCS did not impede the performance of ammonia oxidizing archaea, ammonia oxidizing bacteria (Nitrosomonas) and anammox bacteria (Candidatus Brocadia and Ca. Kuenenia), but significant inhibited the nitrite oxidizing bacteria (Nitrospira and Ca. Nitrotoga) and denitrifying bacteria. The influent TCS led to the increase of EPS content and enrichment of four resistance genes (RGs) (intI1, sul1, mexB, and tnpA), which might be two principal mechanisms by which PN/A can resist TCS. In addition, functional bacteria carrying multiple RGs also contributed to the maintenance of PN/A system function. These findings improved the understandings of antimicrobial effects on the PN/A system.


Interleukin 10, produced in abundance by human newborn T cells, may be the regulator of increased tolerance associated with cord blood stem cell transplantation.
The use of human umbilical cord blood as an alternative source of stem cells to bone marrow for the reconstitution of the immune system is associated with less frequent and less severe incidence of graft-versus-host disease (GVHD). This study focuses on aspects of cord blood T-cell biology that may contribute to a perceived increased tolerance associated with the neonatal immune response. A skewing of the T-helper (Th)1/Th2 phenotype of cord blood T cells towards a Th2 response has frequently been cited as a possible cause. In this study, primary and repeated stimulation via the T-cell receptor (TCR) complex induced a Th0-type cytokine response, with both adult and cord blood-derived naïve T cells producing interferon gamma (IFN-gamma), interleukin 4 (IL-4) and IL-5. IL-10 was induced in cord blood T-cell cultures during primary stimulation, while adult T cells began to secrete IL-10 only after repeated stimulation. The presence of the antigen-presenting cell (APC)-derived cytokine IL-1beta inhibited IL-10 production by cord blood cells. The effects of IL-12 and IL-4 on T-cell cytokine responses were also examined. In addition to their differential Th1/Th2 skewing effects on cord and adult T cells, both cytokines augmented IL-10 production in both T-cell populations. These findings demonstrate that cord blood T cells may secrete large amounts of the anti-inflammatory cytokine IL-10 and that the presence of IL-1beta or Th1/Th2 skewing cytokines can regulate its production. This data provides support for the recognized tolerant nature of the newborn immune response that may contribute to the reduced incidence of GVHD associated with cord blood transplantation.


Subclinical atherosclerosis in patients with systemic lupus erythematosus: A systemic review and meta-analysis.
Systemic lupus erythematosus (SLE) is associated with increased risk of cardiovascular disease. Carotid intima media thickness (CIMT) and carotid plaques are both frequently used to identify populations at higher cardiovascular risk. A systematic literature search and meta-analysis were performed to evaluate CIMT and carotid plaques difference between SLE patients and normal controls. The literatures comparing markers of cardiovascular risk (CIMT and prevalence of carotid plaques) in SLE and controls were systematically searched in PubMed, EMBASE and Cochrane databases. The overall mean CIMT difference and pooled odds ratio (OR) for the prevalence of carotid plaques between SLE patients and control groups were calculated by fixed-effects or random-effect model analysis. Meta-regression was performed to explore the potential influencing factors. Publication bias was examined by a funnel plot and Egger's test. A total of 80 studies (6085 SLE patients and 4794 controls) were included in the final analysis, 71 studies with data on CIMT (4814 cases and 3773 controls) and 44 studies reporting on the prevalence of carotid plaques (4417 cases and 3528 controls). As compared to controls, SLE patients showed a higher CIMT (WMD: 0.07 mm; 95%CI: 0.06, 0.09; P<0.001), and an increased prevalence of carotid plaques (OR: 2.45; 95%CI: 2.02, 2.97; P<0.001). Meta-regression models showed that traditional cardiovascular risk factors (age, HDL and triglyceride of SLE patients) and lupus related risk factors (as expressed by duration, ESR, SLEDAI and steroids) had a significant influence on CIMT, steroids and triglyceride had significant influence on the prevalence of carotid plaques. Our findings support the current evidence base for an increased cardiovascular burden in SLE patients and support the use of CIMT and carotid plaques in observational studies in SLE patients. The findings are of importance to design more specific prevention and treatment strategies.


Modification of gelatin beadlets for zero-order sustained release.
A three-phase suspension process was used for the preparation of gelatin beadlets containing succinylsulfathiazole. When the beadlets were hardened with 10% formalin at 5 degrees C for varying periods of time up to 24 hr, the 6-hr hardening time gave the slowest release rate. Drug release rate from gelatin beadlets was slower in simulated gastric fluid (SGF) than in simulated intestinal fluid (SIF) but the sustained effect was too limited to be useful for most applications. When the hardened gelatin beadlets were coated with cellulose acetate butyrate (CAB) by an emulsion-solvent evaporation method, a more pronounced sustained effect and a nearly zero-order release were found in SIF. The effects of the amount of gelatin used, the amount of CAB employed, and the length of hardening time on drug release were investigated. The treatment of gelatin beadlets with formalin reduced the swelling action of gelatin in aqueous medium. A nonzero-order drug release rate was observed when the gelatin swelled sufficiently to rupture the CAB coating. The drug release rate can be adjusted by using different ratios of hardened gelatin beadlets and CAB coating in which the gelatin enhances the release rate and the CAB serves as a barrier.


Coloration of the Chilean Bellflower, Nolana paradoxa, interpreted with a scattering and absorbing layer stack model.
An absorbing-layer-stack model allows quantitative analysis of the light flux in flowers and the resulting reflectance spectra. It provides insight in how plants can optimize their flower coloration for attracting pollinators. The coloration of flowers is due to the combined effect of pigments and light-scattering structures. To interpret flower coloration, we applied an optical model that considers a flower as a stack of layers, where each layer can be treated with the Kubelka-Munk theory for diffusely scattering and absorbing media. We applied our model to the flowers of the Chilean Bellflower, Nolana paradoxa, which have distinctly different-colored adaxial and abaxial sides. We found that the flowers have a pigmented, strongly scattering upper layer, in combination with an unpigmented, moderately reflecting lower layer. The model allowed quantitative interpretation of the reflectance and transmittance spectra measured with an integrating sphere. The absorbance spectrum of the pigment measured with a microspectrophotometer confirmed the spectrum derived by modeling. We discuss how different pigment localizations yield different reflectance spectra. The absorbing layer stack model aids in understanding the various constraints and options for plants to tune their coloration.


Structure-function relations in the NTPase domain of the antiviral tRNA ribotoxin Escherichia coli PrrC.
Breakage of tRNA by Escherichia coli anticodon nuclease PrrC (EcoPrrC) underlies a host antiviral response to phage T4 infection. Expression of EcoPrrC is cytocidal in yeast, signifying that PrrC ribotoxicity crosses phylogenetic domain boundaries. EcoPrrC consists of an N-terminal NTPase module that resembles ABC transporters and a C-terminal nuclease module that is sui generis. PrrC homologs are prevalent in many other bacteria. Here we report that Haemophilus influenzae PrrC is toxic in E. coli and yeast. To illuminate structure-activity relations, we conducted a new round of mutational analysis of EcoPrrC guided by primary structure conservation among toxic PrrC homologs. We indentify 17 candidate active site residues in the NTPase module that are essential for toxicity in yeast when EcoPrrC is expressed at high gene dosage. Their functions could be educed by integrating mutational data with the atomic structure of the transition-state complex of a homologous ABC protein.


Preventing infections associated with central venous catheters.
CR-BSI is one of the most serious complications in an already seriously ill patient. Incorporation of these recommendations into local protocols and routine clinical practice will help to bring about a significant reduction in the incidence of CR-BSI in all NHS acute care trusts. The evidence base will be reviewed in 2002.


Decreased external skeletal robustness due to reduced physical activity?
Childhood obesity is a global problem, e.g., due to physical inactivity. External skeletal robustness (Frame-Index) has decreased in German schoolchildren. An association between Frame-Index and physical activity was assumed. Further often body mass index (BMI) is analyzed without reference to bone structure. Therefore, we analyze relationships between Frame-Index, BMI, % body fat, and physical activity. In a cross-sectional study, 691 German children aged 6-10 years were investigated. BMI, % body fat, Frame-Index, total steps p.w., sports club rate p.w., training time p.d., and TV-time p.d. were determined. Total steps (P < 0.001), BMI (P < 0.001), and % body fat (P = 0.024) are positively linked to Frame-Index. Total steps (P < 0.001), sports club rate (P = 0.001), and training time (P < 0.001) are negatively associated with % body fat. Total steps (P = 0.017) are negatively linked to BMI. TV-time is positively related to BMI (P < 0.001) and % body fat (P < 0.001). % Body fat is affected by age (P < 0.001), sex (P = 0.028), and total steps (P = 0.002). BMI is influenced by age (P < 0.001), and Frame-Index by sex (P < 0.001) and total steps (P = 0.029). Principal component analysis indicates an association between BMI and TV-time and Frame-Index and total steps. We demonstrate an association between external skeletal robustness and physical activity, which is not captured by in BMI measurements. Children should be physically active in order to maintain skeletal robustness.


TCR-based lineage tracing: no evidence for conversion of conventional into regulatory T cells in response to a natural self-antigen in pancreatic islets.
Foxp3-expressing regulatory T (T reg) cells derive primarily from selection in the thymus. Yet conversion of mature conventional CD4(+) T (T conv) cell lymphocytes can be achieved in several conditions, such as transforming growth factor beta treatment, homeostatic expansion, or chronic exposure to low-dose antigen. Such conversion might provide a means to generate peripheral tolerance by "converting" potentially damaging T cells that react to self-antigens. We tested this hypothesis in mice transgenic for the BDC2.5 T cell receptor (TCR), which is representative of a diabetogenic specificity that is naturally present in NOD mice and reactive against a pancreatic self-antigen. In the thymus, before any exposure to antigen, clonotype-positive T reg and T conv cells express a second TCRalpha chain derived from endogenous loci. High-throughput single-cell sequencing of secondary TCRs of the Valpha2 family showed their joining CDR3alpha regions to be very different in T reg and T conv cell thymocytes. These specific CDR3alpha motifs, thus, provided a "tag" with which to test the actual impact of T conv to T reg cell conversion in response to peripheral self-antigen; should the autoreactive clonotypic TCR induce T conv to T reg cell conversion upon encounter of cognate antigen in the pancreas or draining lymph node, one would expect to detect tag CDR3alpha motifs from T conv cells in the T reg cell populations. Sequencing large numbers of peripheral BDC(+)Valpha2(+) cells showed that little to no conversion occurs in response to this pancreatic autoantigen.


Markedly improved glycemic control and enhanced insulin sensitivity in a patient with type 2 diabetes complicated by a suprasellar tumor treated with pioglitazone and metformin.
A patient with type 2 diabetes and hypothalamic damage due to a suprasellar tumor developed impaired glycemic control and central obesity. The patient showed exaggerated adrenocorticotropic hormone responsiveness as determined by a corticotrophin releasing hormone test and elevated serum leptin concentrations associated with ravenous appetite and insulin resistance mediated in part through disturbances in leptin signaling. Combination treatment with metformin and pioglitazone was markedly effective in improving glycemic control. Additionally, metformin treatment showed marked anorectic effects on the hyperphagia. This case has important implications for the pathogenesis and management of diabetes in patients with hypothalamic-pituitary-adrenal axis deficiencies.


Level of knowledge, acceptability, and willingness to use oral fluid HIV self-testing among medical students in Kilimanjaro region, Tanzania: a descriptive cross-sectional study.
HIV Self-Testing (HIVST) is universally accepted as an innovative strategy complimenting existing HIV testing services to archive the UNAIDS, 95-95-95 goals by 2030. However, the adoption of HIVST is lagging in most sub-Saharan countries, including Tanzania. This study aimed to determine the level of knowledge, acceptability, and willingness to use HIVST among Medical students in Kilimanjaro region, Tanzania. A descriptive cross-sectional study using a self-administered, semi-structured questionnaire was conducted from May to June 2019 among 271 medical students aged 18-44 years enrolled in a degree of Medicine course at Kilimanjaro Christian Medical University College. A total of 271 participants were enrolled (response rate of 100%). The mean age was 23.9 (SD 2.9), the majority (91%) were Christians, being single (92%), and a half (50.2%) were males. More than half (55.7%) was sexually active, 67.5% reported the age of first sexual debut at 19 years and above. The majority (81.5%) reported that they had one sexual partner, 37% used condoms during the last sexual act. The majority (98.7%) had never had a sexually transmitted disease during the past 3 months, 22.5% reported using alcohol when having sex. More than three-quarters (79%) ever tested for HIV, and 41.6% tested for HIV in the past year. More than two-thirds (67.9%) had a high level of knowledge on oral fluid HIV self-test. Being a female was related with high level of knowledge (P = 0.225). The acceptability of HIVST was 62.7%, and about two-thirds showed a willingness to buy a self-test kit if available for public use. The high level of knowledge on oral fluid HIV self-testing, acceptability and willingness to buy self-test kit if available for public use among sexually active medical students underscores the importance of introducing HIVST as a complementary approach for existing HIV testing services in this setting. To make HIVST effective, interventionist should address concerns associated with self-testing, such as lack of pre and post-test counseling, suicidal risks after receiving HIV positive results, perceived risks of inaccurate HIVST test results, lack of linkage to care of individuals receiving HIV positive results, perceived risks of intimate partner violence, coercive testing of a female partner, and perceived high cost of buying self-test kits.


Biomechanical responses of endodontically treated tooth implant-supported prosthesis.
This study was designed to investigate the biomechanical interactions in endodontically treated tooth implant-supported prosthesis with variations in implant system and load type by using the nonlinear finite element (FE) approach. The computed tomography (CT), micro-CT image process, and computer-aided design systems were integrated to construct the FE models containing 1-, 2-, and 3-piece implants splinted to the second premolar. Realistic interface conditions within the implant system were simulated by using frictional contact elements, and mechanical responses in terms of von Mises stress were computed for all models. The simulated results indicated that splinting an endodontically treated tooth to a neighbor implant decreased stress values in dentin and post but increased stress of implant and bone. The oblique occlusal forces increased the stress values relative to those of axial analogs. A splinted system with a 2-piece implant increased stress on the bone and decreased stress on the prosthesis compared with that of the 1-piece implant. Splints with 1-piece implant prosthesis, without eccentric occlusal contacts, have better mechanical responses than those of 2- or 3-piece implants when connected to an endodontically treated tooth restored with post core and crown.


Population-based twin registries: illustrative applications in genetic epidemiology and behavioral genetics from the Finnish Twin Cohort Study.
The population-based twin registries of Denmark, Finland, Norway and Sweden represent an extraordinary resource for scientific research. Although each register has its own history and composition, they share certain common qualities. All the Nordic countries have a long tradition of population registration, a high standard of living and health-related registers of high quality. The large size of the registers means that they are uniquely placed for representative studies of rare occurrences. Examples of studies that these registers make possible are illustrated with data from the Finnish Twin Cohort, which in its first phase consisted of over 17,000 like-sexed twin pairs born before 1958. It has been recently expanded to include multiple births between 1958 and 1986 (nearly 23,000 sets) and their first-degree relatives. During this period the DZ/MZ ratio decreased, while an increase in MZ twinning rates was observed between 1974 and 1986.


Reliability and validity of culturally adapted Turkish Short Musculoskeletal Function Assessment questionnaire (SMFA-TR).
This study aimed to culturally adapt and validate the Turkish version of the Short Musculoskeletal Function Assessment Questionnaire (SMFA-TR) which primarily assesses the functional status of patients. The translation and cross-cultural adaptation of SMFA to Turkish was made by the standardized procedure and tested for clinimetric quality. The following analyses were made to evaluate clinimetric quality of the SMFA-TR: reliability with factor analysis and Chronbach's α (construct validity), correlations between SMFA-TR and Short Form (SF)-36 (concurrent validity), test-retest reliability (intraclass correlation analyses), floor and ceiling effects. The questionnaire was applied to 166 patients with musculoskeletal problems. All patients filled in the SMFA-TR and the validated Turkish SF-36 questionnaire. Forty-two patients returned to complete the same questionnaires at 10 days. Factor analysis revealed a 4-factor structure of the SMFA-TR. Cronbach's α values were over 0.88 for both original subscales (dysfunction and bother) of the SMFA. Internal consistency (0.88-0.94) and test-retest reliability coefficients (0.90-0.98) were high for both subscales. Turkish SF-36 questionnaire conventional subscales showed significant correlations with SMFA-TR subscales. No floor or ceiling effects were found. The Turkish version of the SMFA was found to be reliable and valid for Turkish-speaking patients with musculoskeletal injuries or disorders.


A novel series of N-(hexahydro-1,4-diazepin-6-yl) and N-(hexahydroazepin- 3-yl)benzamides with high affinity for 5-HT3 and dopamine D2 receptors.
A novel series of benzamides with a hexahydro-1,4-diazepine or hexahydroazepine ring in the amine moiety were prepared, and their binding affinities for 5-HT3 and dopamine D2 receptors were evaluated. The R isomer of the 1-ethyl-4-methylhexahydro-1,4-diazepinylbenzamide (R)-22 had potent affinity for both receptors. The R-enantiomer of the corresponding 1-ethylhexahydroazepinylbenzamide 28 showed potent affinity for dopamine D2 receptors with reduced affinity for 5-HT3 receptors, while the S isomer was found to be a potent and selective 5-HT3 receptor antagonist.


Tempo change during body rocking.
Occasional tempo change during spontaneous body rocking is described in two institutionalised mentally retarded children. Both acceleration and deceleration in moment-to-moment rate of body rocking were demonstrated under constant environmental conditions. The findings are discussed in relation to the automatism hypothesis of stereotyped behaviour.


All-trans retinoic acid induces synaptopodin-dependent metaplasticity in mouse dentate granule cells.
Previously we showed that the vitamin A metabolite all-trans retinoic acid (atRA) induces synaptic plasticity in acute brain slices prepared from the mouse and human neocortex (Lenz et al., 2021). Depending on the brain region studied, distinct effects of atRA on excitatory and inhibitory neurotransmission have been reported. Here, we used intraperitoneal injections of atRA (10 mg/kg) in adult C57BL/6J mice to study the effects of atRA on excitatory and inhibitory neurotransmission in the mouse fascia dentata-a brain region implicated in memory acquisition. No major changes in synaptic transmission were observed in the ventral hippocampus while a significant increase in both spontaneous excitatory postsynaptic current frequencies and synapse numbers were evident in the dorsal hippocampus 6 hr after atRA administration. The intrinsic properties of hippocampal dentate granule cells were not significantly different and hippocampal transcriptome analysis revealed no essential neuronal changes upon atRA treatment. In light of these findings, we tested for the metaplastic effects of atRA, that is, for its ability to modulate synaptic plasticity expression in the absence of major changes in baseline synaptic strength. Indeed, in vivo long-term potentiation (LTP) experiments demonstrated that systemic atRA treatment improves the ability of dentate granule cells to express LTP. The plasticity-promoting effects of atRA were not observed in synaptopodin-deficient mice, therefore, extending our previous results regarding the relevance of synaptopodin in atRA-mediated synaptic strengthening in the mouse prefrontal cortex. Taken together, our data show that atRA mediates synaptopodin-dependent metaplasticity in mouse dentate granule cells.


Effect of pedicle fixation combined with (125)I seed implantation for metastatic thoracolumbar tumors.
The aim of this study was to investigate the clinical efficacy of pedicle fixation combined with (125)I brachytherapy in treating metastatic thoracolumbar tumors. A retrospective analysis of the clinical data of seven metastatic thoracolumbar tumor patients who received pedicle fixation combined with radioactive (125)I seed implantation brachytherapy in our department between January 2009 and December 2013 was performed. The visual analog scale (VAS) for pain and the Karnofsky performance status (KPS) score before the operation and 1, 6, and 12 months after the operation were observed and recorded. The changes in the scores at each time point were compared. All the patients underwent a successful operation, without any complications during their hospitalization. All the patients received postoperative follow-up, and the duration of follow-up was 15-50 months, with an average of 32.2 months. One pancreatic cancer patient died of liver failure and hypoproteinemia 28 months post surgery. The VAS scores of patients before the operation and 1, 6, and 12 months after the operation were 7.43±0.98, 2.71±0.49, 3.00±0.82, and 4.29±0.98, respectively; the KPS scores were 52.9±9.5, 84.3±5.3, 75.7±5.3, and 72.9±4.9, respectively. These results suggest that the VAS score at each time point was significantly decreased compared with that before the operation, while the KPS score was significantly increased compared with that before the operation. Both differences had statistical significance (P<0.05). As a therapy for advanced malignant tumors with thoracolumbar metastasis, pedicle fixation combined with (125)I brachytherapy can effectively relieve short-term pain and improve patient's quality of life.


Biologic and psychosocial risk factors of sudden death from coronary disease in white women.
The relation of the risk of sudden death from arteriosclerotic heart disease to psychosocial and biologic factors was studied retrospectively in 64 white women who died suddenly of arteriosclerotic heart disease and in 64 age-related neighborhood control women. All sudden deaths occurred outside the hospital within 24 hours of the onset of symptoms in women who were not incapacitated before death. Women who died suddenly were less often married, exhibited more educational incongruity with their spouses and had fewer children than the control population. Twelve of the 64 women who died suddenly of heart disease and none of the control women had a definite history of psychiatric treatment. The women who died suddenly also smoked more cigarette and consumed greater quantities of alcoholic beverages than age-matched neighborhood control women. Multiple regression analysis revealed that history of psychiatric illness, cigarette smoking, alcohol consumption, educational incongruity and number of children contributed significantly to differences between women who died suddenly of heart disease and control subjects.


TNFAIP8 Promotes Cisplatin Chemoresistance in Triple-Negative Breast Cancer by Repressing p53-Mediated miR-205-5p Expression.
Tumor necrosis factor alpha-induced protein 8 (TNFAIP8) is implicated in the tumor progression and prognosis of triple-negative breast cancer (TNBC), but the detailed regulatory mechanism of TNFAIP8 in cisplatin tolerance in TNBC has not yet been investigated. TNFAIP8 was evidently upregulated in TNBC tumor tissues and cell lines. Knocking down TNFAIP8 led to impaired proliferation and elevated apoptosis of TNBC cells upon cisplatin (DDP) treatment. Mechanistic studies revealed that TNFAIP8 repressed the expression of p53 and p53-promoted microRNA (miR)-205-5p; moreover, miR-205-5p targeted multiple genes required for the cell cycle and repressed Akt phosphorylation, which thus inhibited the proliferation of TNBC cells. In addition, silencing of TNFAIP8 led to the upregulation of miR-205-5p and the restraint of the TRAF2-NF-κB pathway, which thus enhanced the suppressive effects of DDP on tumor growth in nude mice. This study revealed that TNFAIP8 was essential in the DDP tolerance formation of TNBC cells by reducing p53-promoted miR-205-5p expression. Thus, targeting TNFAIP8 might become a promising strategy to suppress TNBC progression.


Two-color femtosecond experiments by use of two independently tunable Ti:sapphire lasers with a sample-and-hold switch.
We performed femtosecond two-color experiments (four-wave mixing and pump probe) using two independently tunable, partially synchronized femtosecond lasers. Despite the fact that the jitter is of the order of 5-10 ps, the time resolution is limited only by the pulse width when a homemade sample-and-hold switch is used.


Radionuclide therapy of hematologic disorders.
32P is effective therapy for polycythemia and primary thrombocytosis. The Polycythemia Vera Study Group is comparing radioactive phosphorus with alkylating agents to determine relative efficacy. Less well investigated is the effectiveness of 32P vs. busulfan in chronic granulocytic leukemia. Endolymphatic administration of radiopharmaceuticals may play a role in the therapy of infradiaphragmatic lymphoma. Among the radionuclides that have at times been used in hematology are 32P, 198Au 24Na, 76As, 89Sr, 52Mb, 54Mn, 91Y, 95Zr, 95Cb, 111Ag, 109Pd, 131I, 185W, and 192Ir. As stated, 32P has proven single most efficacious agent. The hematologic diseases that have been treated include both malignant and benign conditions. Among the malignant conditions are polycythemia vera, agnogenic myeloid metaplasia, thrombocythemia, leukemia, Hodgkin's disease, and multiple myeloma. Hemophilia, and Osler--Weber--Rendu disease are among the benign entities in which the agents have been tried. Polycythemia and thrombocythemia remain those in which the greatest success has been achieved.


Non-equilibrium degassing and a primordial source for helium in ocean-island volcanism.
Radioactive decay of uranium and thorium produces 4He, whereas 3He in the Earth's mantle is not produced by radioactive decay and was only incorporated during accretion-that is, it is primordial. 3He/4He ratios in many ocean-island basalts (OIBs) that erupt at hotspot volcanoes, such as Hawaii and Iceland, can be up to sixfold higher than in mid-ocean ridge basalts (MORBs). This is inferred to be the result of outgassing by melt production at mid-ocean ridges in conjunction with radiogenic ingrowth of 4He, which has led to a volatile-depleted upper mantle (MORB source) with low 3He concentrations and low 3He/4He ratios. Consequently, high 3He/4He ratios in OIBs are conventionally viewed as evidence for an undegassed, primitive mantle source, which is sampled by hot, buoyantly upwelling deep-mantle plumes. However, this conventional model provides no viable explanation of why helium concentrations and elemental ratios of He/Ne and He/Ar in OIBs are an order of magnitude lower than in MORBs. This has been described as the 'helium concentration paradox' and has contributed to a long-standing controversy about the structure and dynamics of the Earth's mantle. Here we show that the helium concentration paradox, as well as the full range of noble-gas concentrations observed in MORB and OIB glasses, can self-consistently be explained by disequilibrium open-system degassing of the erupting magma. We show that a higher CO2 content in OIBs than in MORBs leads to more extensive degassing of helium in OIB magmas and that noble gases in OIB lavas can be derived from a largely undegassed primitive mantle source.


Implementation of a centralized discharge planning office during the COVID-19 pandemic: translating the experience from the emergency to routine clinical practice.
Background and aim During the COVID-19 emergency, the lombardy region (northern Italy) implemented a regional Centralized Discharge Planning Office (CDPO) to promptly manage the discharge requests, rapidly match the needs of discharge hospitals with the availability of admission facilities and ensure the management of the entire discharge process. To improve the discharge process in routine clinical practice, maintaining the role of the CDPO could be of great interest. This paper describes the experience of the CDPO during the COVID-19 pandemic and discusses the possibility to translate this operational model to routine clinical practice. The PRIAMO web portal was developed to manage discharge requests with centralized and standardized procedures. The activity on PRIAMO consisted of three stages: discharge request, sorting process, and discharge follow-up phase. To evaluate the activity of the CDPO, these indicators were considered: average time (hours) between patient discharge and transfer acceptance; average time (hours) between patient discharge and effective admission to the new facility; percentage of transfers whose destination was found directly by the CDPO; percentage of reallocations beyond 24 hours; mean distance between discharge and admission facilities. Process indicator evaluation showed a great reduction in the time between the discharge and the admission to post-acute care facilities. Transfers whose destination was found directly by the CDPO progressively increased. Reallocations beyond 24 hours by the CDPO decreased, suggesting an improvement in the quality of the operations. Centralized discharge planning has enabled timely and efficient management of discharge requests even in the moment of a surge, saving time and costs for acute care hospitals.


Schwann cell remyelination of demyelinated axons in spinal cord multiple sclerosis lesions.
To investigate remyelination in multiple sclerosis lesions, we immunostained spinal cord sections from patients with multiple sclerosis and neurological normal (control) patients with antisera to P0 protein, a major constituent of peripheral nervous system myelin, and myelin basic protein, which is found in both central and peripheral nervous system myelin. In sections from five of the eight patients with no clinical or pathological evidence of neurological disease, P0 immunostaining was confined to peripheral myelin sheaths in dorsal and ventral roots. They were intensely stained, and peripheral--central nervous system transition zones were clearly demarcated. Sections from the other three control patients contained a few P0-stained sheaths in the central nervous system near root entry zones or among marginal glia near the dorsal sulcus. Spinal cord sections from six of the ten patients with multiple sclerosis contained clusters of myelin sheaths immunostained by P0 antiserum. These regenerating sheaths of peripheral nervous system origin were most numerous in large lesions and were commonly located in central areas or peripherally near root entry zones. The sheaths were observed frequently in areas of active demyelination and appeared morphologically normal even when surrounded by debris-filled macrophages. Near margins of small inactive plaques were a few basic protein--stained oligodendroglia extending processes to thin basic protein--stained sheaths.(ABSTRACT TRUNCATED AT 250 WORDS)


The sites of cleavage in oxidized insulin-B chain by a hemorrhagic protease derived from the venom of the habu (Trimeresurus flavoviridis).
The substrate specificity of a hemorrhagic protease derived from habu (Trimeresurus flavoviridis) venom was determined using oxidized insulin-B chain as a substrate. The enzyme was shown to be a fairly specific protease; it cleaved the substrate essentially at two positions, His10-Leu11 and Ala14-Leu15. The sites cleaved with the protease were compared to those reported with hemorrhagic proteases from other snake venoms.


Reliability and validity of Kannada version of Victorian Institute of Sports Assessment for patellar tendinopathy (VISA-P-K) questionnaire.
In sporting activities, patellar tendon is commonly subjected to overuse and it influences the athletes' career especially in elite basketball and volleyball players. A condition specific and self-administrated questionnaire is required for clinical and research purpose to rate disability. The Victorian Institute of Sports Assessment for patellar tendinopathy (VISA-P) questionnaire is widely accepted and a valid tool to assess pain and athletes, suffering from PT, ability to participate in sports. So, the objective of this study is to develop and validate Kannada version of VISA-P questionnaire among Kannada speaking population athletes. To translate VISA-P questionnaire into Kannada and to study reliability and validity of Kannada version. Translation and cross-culture adaptation were performed according to the guidelines followed internationally. Those are: translation, synthesis, back-translation, revision by expert committee, pretesting and appraisal by advisory committee. The psychometric properties were tested by administering the questionnaire to 70 subjects (35 symptomatic and 35 asymptomatic). For test-retest reliability, athletes with patellar tendinopathy were tested twice within a week and test-retest reliability was assessed by Intra-class correlation coefficient (ICC). The validity was tested by comparing the VISA-P-Kannada questionnaire (VISA-P-K) against Blazina classification of patellar tendinopathy (BCPT) using Spearman correlation coefficient. The test-retest reliability was excellent (ICC 0.97). Internal consistency was found to be good (Cronbach's alpha = 0.98). The VISA-PK showed significant correlation by comparing against Blazina and Spearman's score (r = 0.72, p < 0.01). The VISA-P-K translation and adaptation processes were successful. The adapted instrument demonstrated good psychometric properties. The VISA-P-K has proven to be reliable and valid tool and can be used in the Kannada speaking patients with patellar tendinopathy.


Aquatic photochemistry of Cu(II) in the presence of As(III): Mechanistic insights from Cu(III) production and As(III) oxidation under neutral pH conditions.
Surface complexation between arsenite (As(III)) and colloidal metal hydroxides plays an important role not only in the immobilization and oxidation of As(III) but also in the cycle of the metal and the fate of their ligands. However, the photochemical processes between Cu(II) and As(III) are not sufficiently understood. In this work, the photooxidation of As(III) in the presence of Cu(II) under neutral pH conditions was investigated in water containing 200 μM Cu(II) and 5 μM As(III) under simulated solar irradiation consisting of UVB light. The results confirmed the complexation between As(III) and Cu(II) hydroxides, and the photooxidation of As(III) is attributed to the ligand-to-metal charge transfer (LMCT) process and Cu(III) oxidation. The light-induced LMCT process results in simultaneous As(III) oxidation and Cu(II) reduction, then produced Cu(I) undergoes autooxidation with O2 to produce O2•⁻ and H2O2, and further the Cu(I)-Fenton reaction produces Cu(III) that can oxidize As(III) efficiently (kCu(III)+As(III) = 1.02 × 109 M-1 s-1). The contributions from each pathway (ρrCu(II)-As(III)+hv = 0.62, ρrCu(III)+As(III) = 0.38) were obtained using kinetic analysis and simulation. Sunlight experiments showed that the pH range of As(III) oxidation could be extended to weak acidic conditions in downstream water from acid mine drainage (AMD). This work helps to understand the environmental chemistry of Cu(II) and As(III) regarding their interaction and photo-induced redox reactions.


Fat absorption in preterm infants fed a taurine-enriched formula.
An adapted cow's milk infant formula without or with extra taurine (350 mumol/l) was fed to four and five infants, respectively. The infants, born after 28-32 weeks gestation, and initially fed with a starting formula for preterms, were switched to one of the two above-mentioned formulae at approximately the 16th day of life. Each infant was studied during 4 consecutive weeks. The faecal excretion of fat, energy and total bile acids was determined from 3-day stool collections each week. The addition of taurine to the infant formula neither improved the uptake of fat and energy nor changed the faecal bile acid excretion. Growth velocity was similar in both groups of infants. Based on these results there is no rationale for adding taurine to adapted cow's milk infant formula to obtain a better fat absorption.


[Progressive pulmonary fibrosis during combination chemotherapy with BCNU (author's transl)].
In two patients with acute leukaemia, the development of progressive interstitial pulmonary fibrosis was observed following chemotherapy with BCNU, Cytoxan and Ara-C. The x-ray changes were accompanied by restrictive changes of pulmonary function and, later on, by severe hypoxia. Serologic tests did not reveal infection with cytomegaly virus or mycoplasma pneumoniae. These findings, together with reports in the literature, suggest a toxic effect of BCNU on the lung. The combination with Cyclophosphamide may contribute to this toxic reaction.


Pharmacophore modeling coupled with scaffold hopping to identify novel and potent ribosomal S6 kinase (RSK2) protein antagonists as anti-cancer agents.
Communicated by Ramaswamy H. Sarma.


On the encapsulation of Olsalazine by β-cyclodextrin: A DFT-based computational and spectroscopic investigations.
In this work, the supramolecular host-guest interaction of the prodrug Olsalazine (OLZ) and β-Cyclodextrin (β-CD) was examined experimentally and computationally. Experimentally, employing the UV-Vis spectroscopic method in aqueous media at various pH's, results obtained using the Benesi-Hilderbrand approach demonstrated that OLZ can form supramolecular inclusion complex with β-CD with stoichiometric ratio of 1:1. Furthermore, these results revealed that the formation of OLZ: β-CD complexes exhibited insignificant pH dependency in the range 5-8 with an average binding constant (Kb) of approximately 1×103M-1. Computationally, geometry optimization of 1:1 OLZ: β-CD complexes was performed employing the ONIOM (DFT((ωB97XB)/6-31+G(d)),SQM(PM3)) approach. Obtained results demonstrated that OLZ: β-CD complex is stabilized by the formation of intermolecular hydrogen bonds with an average length of approximately 1.8Å. Additionally, the stability of OLZ: β-CD complex was demonstrated employing ADMP molecular dynamic simulations over a timeframe of 500fs. The molecularity of the supramolecular host-guest interaction between OLZ and β-CD is presented and interpreted in the essence of TD-DFT and molecular orbitals analyses.


Association of sleep bruxism with ceramic restoration failure: A systematic review and meta-analysis.
Ceramic restorations are popular because of their excellent optical properties. However, failures are still a major concern, and dentists are confronted with the following question: is sleep bruxism (SB) associated with an increased frequency of ceramic restoration failures? The purpose of this systematic review and meta-analysis was to assess whether the presence of SB is associated with increased ceramic restoration failure. Observational studies and clinical trials that evaluated the short- and long-term survival rate of ceramic restorations in SB participants were selected. Sleep bruxism diagnostic criteria must have included at least 1 of the following: questionnaire, clinical evaluation, or polysomnography. Seven databases, in addition to 3 nonpeer-reviewed literature databases, were searched. The risk of bias was assessed by using the meta-analysis of statistics assessment and review instrument (MAStARI) checklist. Eight studies were included for qualitative synthesis, but only 5 for the meta-analysis. Three studies were categorized as moderate risk and 5 as high risk of bias. Clinical and methodological heterogeneity across studies were considered high. Increased hazard ratio (HR=7.74; 95% confidence interval [CI]=2.50 to 23.95) and odds ratio (OR=2.52; 95% CI=1.24 to 5.12) were observed considering only anterior ceramic veneers. Nevertheless, limited data from the meta-analysis and from the restricted number of included studies suggested that differences in the overall odds of failure concerning SB and other types of ceramic restorations did not favor or disfavor any association (OR=1.10; 95% CI=0.43 to 2.8). The overall quality of evidence was considered very low according to the GRADE criteria. Within the limitations of this systematic review, the overall result from the meta-analysis did not favor any association between SB and increased odds of failure for ceramic restorations.


B-type natriuretic peptide assays for detecting heart failure in the elderly: same value as those in the younger?
Establishing a diagnosis of heart failure in elderly patients is notoriously difficult, especially in those who experience slowly progressive breathlessness, have multiple co-morbidity, and when, as in primary care echocardiography is not routinely (or readily) available. B-type natriuretic peptides measurements are useful in the diagnostic assessment of such patients, although the evidence in the very old (i.e. those aged 75 years and over) is less clear than for younger patients. Conflicting results in literature regarding B-type natriuretic peptides are mainly caused by differences in assays, the applied 'gold standard' for heart failure, and the population studied. Moreover, there is no consensus as to whether similar cut-off points should be applied across age-groups. Numerous studies showed that plasma levels of B-type natriuretic peptide are elevated in the elderly, including the 'healthy' ones. Age-related myocardial fibrosis and subtle diastolic dysfunction that are not detectable by current techniques, and reduced renal clearance have been suggested as reasons for this phenomenon. Importantly, B-type natriuretic peptides are not specific for heart failure, but reflect haemodynamic myocardial stress independent of the underlying pathology, and thus are more 'markers' of the general cardiac state. Cut-points differ widely when comparing studies in patients with acute versus chronic dyspnoea. In patients with acute dyspnoea age-dependent cut-points should be used, while in patients with chronic dyspnoea (that is, slowly progressive breathlessness) at least thresholds below which heart failure can be excluded seem rather independent of age. Especially in the primary care setting where elderly patients with slowly progressive dyspnoea are investigated, the excellent exclusionary capacities of B-type natriuretic peptides are of great help to select those that require echocardiography to establish or eventually reject a diagnosis of heart failure.


Synergistic effect of PAF inhibition and X-ray irradiation in non-small cell lung cancer cells.
Proliferating cell nuclear antigen-associated factor (PAF) is involved in cancer cell growth and associated with cell death induced by ultraviolet (UV) radiation. However, the contribution of PAF to radiotherapy sensitivity in non-small cell lung cancer (NSCLC) is unknown. The aim of this study was to investigate the relationship between PAF expression and radiotherapy response in NSCLC. Associations between PAF expression and patient survival outcomes were evaluated using publicly available online gene expression datasets. RNA interference was performed to knockdown PAF expression in the NSCLC cells. The effects of PAF knockdown on cell proliferation, migration, apoptosis, DNA damage, and activation of MEK/ERK and Wnt/β-catenin signaling pathways following X‑ray irradiation were evaluated in vitro. PAF was found to be overexpressed in lung cancer tissues compared with normal samples, and elevated PAF expression was significantly correlated with inferior patient survival. In vitro, knockdown of PAF inhibited cell proliferation, cell apoptosis, and migration induced by X‑ray irradiation. Moreover, X‑ray-induced intracellular DNA strand damage was more obvious following PAF knockdown. Additionally, PAF knockdown inhibited activation of the MEK/ERK and Wnt/β-catenin signaling pathways in X‑ray-irradiated A549 cells. These data demonstrate that reduced expression of PAF enhances radiosensitivity in NSCLC cells. Mechanistically, inhibition of the MEK/ERK and Wnt/β-catenin signaling pathways caused by PAF interference may lead to impaired cell function and enhance sensitivity to X‑rays. Targeting PAF may therefore serve as a potential therapeutic strategy to increase the efficiency of radiotherapy in NSCLC patients, ultimately improving patient survival.


[Interorgan metabolic correlations during muscular activity and fatigue].
Biochemical and mathematical analysis revealed that metabolic processes in muscles, blood, liver, and brain during both unfatiguing muscular activity and in fatigue change their correlative bonds not identealby. The experiments exposed as a common traits, as differences in metabolic structure of both rapidly and slowly developed fatigue.


Impaired oxidation of carbon-labeled galactose by alcoholic or diabetic liver in vivo.
Because of the organ and enzyme specificity of the metabolism of galactose, evaluation of various kinds of liver disease can be done by measuring the formation of labeled breath CO2 from carbon-labeled galactose in vivo. As shown earlier with uniformly 14C- or 13C-labeled galactose, a further study of alcoholic cirrhotic patients and controls with cheaper 1-14C-galactose indicates a superior discriminatory value of this test compared with common liver function tests. The oxidation test is easier to perform and more acceptable to patients than the standard galactose tolerance blood test. Output of 14CO2 showed slight correlations with serum albumin and 99mTc-sulfur colloid scan grade, but not with other function tests (SGOT, alkaline phosphatase, bilirubin). Comparison with five-year clinical outcome (two groups: with or without known liver-related death) in 29 of 43 total cirrhotic patients (U-14C or 1-14C-galactose) showed a low (75% probability) significance of prognosis for the galactose oxidation test, but none for any of the other tests. A two-part test of oxidation of 14C-galactose (with and without an acute dose of ethanol) in 19 possibly or likely alcoholic (but non-cirrhotic) persons indicated, by correlation with other liver function tests and drinking history, some possibly enhanced sensitivity of the two-part versus the single test for recognizing early liver damage. A preliminary study of the single galactose oxidation test in 7 patients with Type II diabetes suggests moderate impairment of oxidation, which might be applied to evaluate the hepatic disorder in diabetes.


Spatiotemporal brain signal associated with high and low levels of proactive motor response inhibition.
Proactive motor response inhibition is used to strategically restrain actions in preparation for stopping. In this study, we first examined the event related potential (ERP) elicited by low and high level of proactive response inhibition, as assessed by the stop-signal task. Corroborating previous studies, we found an increased amplitude of the contingent negative variation (CNV) in the high level of proactive inhibition. As the main goal of the present study, swLORETA was used to determine the neural generators characterising CNV differences between low and high levels of proactive inhibition. Results showed that the higher level of proactive inhibition involved numerous generators, including within the middle and medial frontal gyrus. Importantly, we observed that the lower level of proactive inhibition also involved a specific neural generator, within the frontopolar cortex. Altogether, present findings identified the specific brain sources of ERP signals involved in the later phase of motor preparation under low or high levels of proactive motor response inhibition.


Ivermectin's Role in the Prevention of COVID-19: A Systematic Review and Meta-Analysis.
This systematic review was performed to determine the population that benefited from prophylactic ivermectin. Seven databases of health-related studies were searched for eligible trials without language restrictions. Randomized controlled trials (RCTs) and cohort studies investigating ivermectin for coronavirus disease 2019 (COVID-19) prevention were included. Data were pooled using a random-effects model, and subgroups were analyzed by study type and the pre- or postexposure population. The certainty of the evidence was determined by the Grading of Recommendations Assessment, Development, and Evaluation approach. Furthermore, 4 RCTs and 4 cohort studies with a moderate to high risk of bias were included in the analysis. The prophylactic use of ivermectin significantly decreased the overall incidence of COVID-19 (odds ratio [OR], 0.26; 95% confidence interval [CI], 0.16-0.44). Nevertheless, the positive result was not supported by the RCT. Ivermectin was associated with a lower risk of COVID-19 (OR, 0.22; 95% CI, 0.12-0.40) in the preexposure population, whereas no protective effect was observed in the postexposure population (OR, 0.39; 95% CI, 0.09-1.67). In summary, prophylactic ivermectin did not prevent COVID-19 in the postexposure population. Although the protective effect of ivermectin was shown in the overall and preexposure populations, the results were unreliable owing to poor-quality evidence.


Pathological analyses of early recurrence and malignant transformation in meningiomas.
We investigated factors of the early recurrence and malignant transformation of histologically benign meningiomas using immunohistochemistry for MIB-1 positive indices (PI) and p53 protein expression, a flow cytometric DNA analysis, and the examination of numerical chromosomal aberrations detected by fluorescence in situ hybridization using an alpha-satellite DNA probe and a bcr gene locus-specific probe. Twenty-six meningiomas of 23 patients were classified into two groups: the 3 patients in whom a recurrence was defined within two years after initial surgery and who showed histologically malignant features were classified as the early recurrent group, and the other 20 patients in whom recurrence did not develop during the same period were classified as the nonrecurrent group. DNA aneuploidy was observed in 40% of the nonrecurrent patients and in 67% of the early recurrent patients. Loss of chromosome 22 was the most common numerical aberration, but the aberrations characteristic of early recurrent meningiomas were not detected. The MIB-1 PI values of the early recurrent meningiomas were higher than those of nonrecurrent meningiomas, suggesting that MIB-1 PI is very important for biological and histopathological analyses and prediction of the future recurrence of meningiomas.


Diabetes Mellitus as a Risk Factor for Acute Postoperative Complications Following Elective Adult Spinal Deformity Surgery.
Retrospective cohort study. Diabetes mellitus is a highly prevalent disease in the United States. Adult spinal deformity (ASD) surgery encompasses a wide variety of spinal disorders and is associated with a morbidity rate between 20% and 80%. Considering utilization of spinal surgery will continue to increase, this study investigates the influence of diabetes mellitus on acute postoperative outcomes following elective ASD surgery. The 2010-2014 American College of Surgeon's National Surgical Quality Improvement Program database was queried using Current Procedural Terminology and International Classification of Diseases (9th Revision) diagnosis codes relevant to ASD surgery. Patients were divided into cohorts based on their diabetic status. Bivariate and multivariate logistic regression analyses were employed to identify which 30-day postoperative outcomes patients are at risk for. A total of 5809 patients met the inclusion criteria for the study of which 4553 (84.2%) patients were nondiabetic, 578 (10.7%) patients had non-insulin-dependent diabetes mellitus (NIDDM), and 275 (5.1%) patients had insulin-dependent diabetes mellitus (IDDM). Diabetes status was significantly associated with length of stay ≥5 days (NIDDM: odds ratio [OR] = 1.27, 95% confidence interval [CI] = 1.02-1.58, P = .034; IDDM: OR = 1.55, 95% CI = 1.15-2.09, P = .004), any complication (NIDDM: OR = 1.26, 95% CI = 1.01-1.58, P = .037), urinary tract infection (NIDDM: OR = 1.87, 95% CI = 1.14-3.05, P = .012), and cardiac complications (IDDM: OR = 4.05, 95% CI = 1.72-9.51, P = .001). Given the prevalence of diabetes, surgeons will invariably encounter these patients for ASD surgery. The present study identifies the increased risk NIDDM and IDDM patients experience following ASD surgery. Quantification of this increased risk may improve the selection of appropriate surgical candidates, patient risk stratification, and patient postoperative safety.


Scale up fractionation of components from novel glabrous brown and yellow canary seeds (Phalaris canariensis L.) and techno-functional properties of the resulting protein isolates.
Glabrous canary seed (Phalaris canariensis L.) is a novel true cereal grain produced primarily in Western Canada which has been approved for human consumption by the U.S. Food and Drug Administration and Health Canada in 2016. Due to its high protein content (22%), this new edible grain is emerging as an alternative source of plant proteins. In the present work, protein extractability from four novel glabrous (2 yellow and 2 brown) canary seeds varieties was improved based on the selection of optimal pH of protein solubilisation and precipitation. Solubilisation at pH 12 followed by acid precipitation at pH 5 were retained as optimal conditions. Scale up of the protein optimized wet fractionation process resulted in highly purified canary seed protein isolates (purity of 91 to 93%) with protein recovery yield of 65 to 69%. In parallel, for the others canary seed components, a good recovery yields were obtained for the oil fraction (6.1-6.7 g/100 g flour), starch fraction (48.1-54 g/100 g flour), and crude fiber fraction (15.1-19.7 g/100 g flour). The study of the functional properties of the obtained canary protein isolates revealed, higher solubility at acidic than alkaline region; enhanced fat and water holding capacities and notably higher foaming and emulsifying capacities than control soy protein isolate. With growing global demand for protein, glabrous canary seed has high potential in the food industry, particularly as a good source of functional gluten free cereal proteins.


Population processing--a powerful class of parallel algorithms.
We present a model of optimization of cost functions by a population of parallel processors and argue that especially diploid recombination of gene strings is a promising recipe for optimization which nature proliferates. Based on a simulated evolutionary search strategy diploidy is introduced as a means for maintaining variability in computational problems with large numbers of local extrema. A differentiation into genotypes and phenotypes is performed. The applied strategy is compared to some traditional algorithms simulating evolution on the basis of two sample cost functions.


Integrity of lichen cell membranes as an indicator of heavy-metal pollution levels in soil.
The epigeic lichens Cladonia rei and Diploschistes muscorum are effective heavy-metal-tolerant colonisers of highly polluted and disturbed sites. In this study we compare their bioaccumulation capacities, accumulation patterns, and responses to heavy-metal stress, as expressed in terms of cell membrane damage. We also aim at verifying the relationships between cell membrane damage and levels of soil pollution with heavy metals, and thereby to identify the bioindicative value of this physiological parameter. Total and intracellular concentrations of Zn, Pb, Cd, As, Cu, and Ni were measured in 140 samples of lichens and corresponding soil, collected from variously contaminated sites. Relative electrical conductivity (EC%) values were determined concurrently in the lichen samples. The studied lichens differ considerably in intracellular uptake susceptibility and the related reduction in membrane integrity. In C. rei thalli, more than half of Zn, Pb, Cd, and As loads are accumulated extracellularly, whereas D. muscorum exhibits a tendency towards intracellular accumulation of the same elements. This property is clearly reflected in cell membrane damage, which is considerably greater in the latter species irrespective of study site. This indicates that intracellular heavy-metal accumulation affects the level of cell membrane damage. Two soil pollution classes were distinguished for both lichens based on element contents in host-substrate samples. The losses of cell membrane integrity in lichen thalli are related to these classes. EC% values above 16 in C. rei and above 20 in D. muscorum suggest elevated levels of heavy metals in the soil. Consequently, this physiological parameter can serve as an early warning indicator for detection of elevated metal concentrations in soil. The biomonitoring method proposed here involves common and widespread lichen species and can be widely applied in post-industrial areas.


Species differences, influence of dose and application on biotransformation of phenol in fish.
1. The metabolism of phenol in goldfish (Carassius auratus), rainbow trout (Salmo gairdneri) and goldenorfe ide (Leuciscus ideus melanotus) was compared. All three species produced phenyl sulphate, quinol sulphate and phenyl glucuronide. 2. The dose dependence of metabolism of phenol in goldfish was investigated. When the concn. of phenol was increased from 0.2 to 2 mg/l medium, that glucuronylated increased from 7 to 16% and that sulphated decreased from 63 to 47%. 3. The influence of mode of exposure on metabolism of phenol in goldfish was examined. Dietary exposure and uptake from medium was compared with i.p. injection. There was less metabolism in the order: dietary exposure greater than uptake from medium greater than injection.


Inhibition of the kinase ITK in a mouse model of asthma reduces cell death and fails to inhibit the inflammatory response.
Interleukin-2 (IL-2)-inducible T cell kinase (ITK) mediates T cell receptor (TCR) signaling primarily to stimulate the production of cytokines, such as IL-4, IL-5, and IL-13, from T helper 2 (TH2) cells. Compared to wild-type mice, ITK knockout mice are resistant to asthma and exhibit reduced lung inflammation and decreased amounts of TH2-type cytokines in the bronchoalveolar lavage fluid. We found that a small-molecule selective inhibitor of ITK blocked TCR-mediated signaling in cultured TH2 cells, including the tyrosine phosphorylation of phospholipase C-γ1 (PLC-γ1) and the secretion of IL-2 and TH2-type cytokines. Unexpectedly, inhibition of the kinase activity of ITK during or after antigen rechallenge in an ovalbumin-induced mouse model of asthma failed to reduce airway hyperresponsiveness and inflammation. Rather, in mice, pharmacological inhibition of ITK resulted in T cell hyperplasia and the increased production of TH2-type cytokines. Thus, our studies predict that inhibition of the kinase activity of ITK may not be therapeutic in patients with asthma.


Characteristics of Cadmium Sorption by Heat-Activated Red Mud in Aqueous Solution.
Red mud as a waste material is produced in large quantities by the aluminum industry. Heat activation has been used to enhance sorption capacity of red mud for its beneficial reuse as an effective sorbent. In this study, heat-activated red mud (HARM) was investigated for its Cd(II) sorption capacity under various process conditions (Cd concentration, pH and contact time) using response surface methodology (RSM). Analysis with RSM identified pH as the most important process parameter. The positive correlation between higher pH and greater Cd(II) sorption was likely due to: (i) decreased proton competition with Cd(II) for sorption sites at higher pH; (ii) enhanced sorption via ion exchange by monovalent Cd species from hydrolysis at higher pH; and (iii) improved thermodynamics of sorption at higher pH as protons are being released as products. Further analysis indicated the sorption process was thermodynamically favorable with a negative change in Gibbs free energy. Additionally, the sorption process exhibited a positive change in enthalpy, indicative of endothermic nature of sorption; this is consistent with sorption increase at higher temperature. These findings provide needed insight into the mechanisms underlying Cd(II) sorption by HARM for more effective applications of heat-activated red mud as sorbents for Cd(II) removal.


Costovertebral Osteoarthrosis: Rare Differential Diagnosis of Back pain in young patients. Case Report.
The differential diagnosis of dorsal thoracic pain can be a challange due to the proximity of the dorsal column to vital organs as well as to its unique anatomy, innervation, and rib joint. The patterns of referred visceral pain require, in most cases, extensive complementary diagnostic tests in order to exclude severe conditions. Referred pain patterns often result in numerous and expensive visceral workups in order to exclude serious conditions, and costovertebral joint osteoarthritis is usually only considered when the origin of the pain remains unexplained. The authors present the case of a 40-year-old man with disabling dorsal pain due to isolated costovertebral osteoarthrosis. The symptomatology was controlled after injection of methylprednisolone guided by computed tomography. This clinical case aims to describe the clinical presentation of a rare entity that should be considered in the differential diagnosis of back pain.


Sex steroids do not influence somatic growth in childhood.
The influence of sex steroids on somatic growth during childhood was evaluated by reviewing linear growth characteristics of 18 agonadal patients with normal sex chromosomes. None of the heights throughout childhood and before the onset of sex steroid therapy were below 2 SDs of the mean. Based on the normal z scores of these patients, we concluded that somatic growth throughout the childhood and prepubertal years is not sex-steroid dependent.


State-of-the-art review of the application and development of various methods of aerosol therapy.
Aerosol therapy is a rapidly developing field of science. Due to a number of advantages, the administration of drugs to the body with the use of aerosol therapy is becoming more and more popular. Spraying drugs into the patient's lungs has a significant advantage over other methods of administering drugs to the body, including injection and oral methods. In order to conduct proper and effective aerosol therapy, it is necessary to become familiar with the basic principles and applications of aerosol therapy under various conditions. The effectiveness of inhalation depends on many factors, but most of all on: the physicochemical properties of the sprayed system, the design of the medical inhaler and its correct application, the dynamics of inhalation (i.e. the frequency of breathing and the volume of inhaled air). It is worth emphasizing that respiratory system diseases are one of the most frequently occurring and fastest growing diseases in the world. Accordingly, in recent years, a significant increase in the number of new spraying devices and pharmaceutical drugs for spraying has appeared on the market. It should also be remembered that the process of spraying a liquid is a complicated and complex process, and its efficiency is very often characterized by the use of micro- and macro parameters (including average droplet diameters or the spectrum of droplet diameter distribution). In order to determine the effectiveness of the atomization process and in the delivery of drugs to the patient's respiratory tract, the analysis of the size of the generated aerosol droplets is most often performed. Based on the proposed literature review, it has been shown that many papers dealt with the issues related to aerosol therapy, the selection of an appropriate spraying device, the possibility of modifying the spraying devices in order to increase the effectiveness of inhalation, and the possibility of occurrence of certain discrepancies resulting from the use of various measurement methods to determine the characteristics of the generated aerosol. The literature review presented in the paper was prepared in order to better understand the spraying process. Moreover, it can be helpful in choosing the right medical inhaler for a given liquid with specific rheological properties. The experimental data contained in this study are of great cognitive importance and may be of interest to entities involved in pharmaceutical product engineering (in particular in the case of the production of drugs containing liquids with complex rheological properties).


"I hated those darn chickens...": the power in stories for older adults and nurses.
Attention to stories provides a holistic view of an older adult and offers the potential for quality nursing intervention. Explored in this article is the power of stories to advance the well being of older adults and the implications for gerontological nurses of a commitment to support storytelling in this population.


Free fatty acid receptor 1 stimulates cAMP production and gut hormone secretion through Gq-mediated activation of adenylate cyclase 2.
Free fatty acid receptor 1 (FFAR1) is highly expressed in enteroendocrine cells of the small intestine and pancreatic beta cells, where FFAR1 agonists function as GLP-1 and insulin secretagogues, respectively. Most efficacious are so-called second-generation synthetic agonists such as AM5262, which, in contrast to endogenous long-chain fatty acids are able to signal through both IP3/Ca2+ and cAMP pathways. Whereas IP3 signaling is to be expected for the mainly Gq-coupled FFAR1, the mechanism behind FFAR1-induced cAMP accumulation remains unclear, although originally proposed to be Gs mediated. When stimulated with AM5262, we observe that FFAR1 can activate the majority of the Gα proteins, except - surprisingly - members of the Gs family. AM5262-induced FFAR1-mediated transcriptional activation through cAMP response element (CREB) was blocked by the specific Gq inhibitor, YM253890. Furthermore, in Gq-deficient cells no CREB signal was observed unless Gq or G11 was reintroduced by transfection. By qPCR we determined that adenylate cyclase 2 (Adcy2) was highly expressed and enriched relative to the nine other Adcys in pro-glucagon expressing enteroendocrine cells. Co-transfection with ADCY2 increased the FFAR1-induced cAMP response 4-5-fold in WT HEK293 cells, an effect fully inhibited by YM253890. Moreover, co-transfection with ADCY2 had no effect in Gq-deficient cells without reintroduction of either Gq or G11. Importantly, although both AM5262/FFAR1 and isoproterenol/β2 adrenergic receptor (β2AR) induced cAMP production was lost in Gs-deficient cells, only the β2AR response was rescued by Gs transfection, whereas co-transfection with ADCY2 was required to rescue the FFAR1 cAMP response. In situ hybridization demonstrated a high degree of co-expression of ADCY2 and FFAR1 in enteroendocrine cells throughout the intestine. Finally, in the enteroendocrine STC-1 and GLUTag cell lines AM5262-induced cAMP accumulation and GLP-1 secretion were both blocked by YM253890. Our results show that Gq signaling is responsible not only for the IP3/Ca2+ but also the cAMP response, which together are required for the highly efficacious hormone secretion induced by second-generation FFAR1 agonists - and that ADCY2 presumably mediates the Gq-driven cAMP response.


Antipsychotic treatment reduces psychotic symptoms and markers of low-grade inflammation in first episode psychosis patients, but increases their body mass index.
The main goal of the present study was to analyze levels of cytokines of the interleukin family (IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-8 and IL-10), interferon-gamma (IFN-γ), monocyte chemoattractant protein-1 (MCP1), tumor necrosis factor-alpha (TNF-α), and vascular endothelial and endothelial growth factors (VEGF and EGF), in the blood samples of first-episode psychosis (FEP) patients before and seven months after the start of antipsychotic medication use. 38 anti-psychotic medication-naïve FEP patients and 37 healthy controls (HC) were recruited. Biochip array technology was used to measure cytokines and growth factors. The comparison of these markers in FEP patients and HC revealed significantly higher levels of EGF, IL-4 and IL-6 and significantly lower level of IL-1β in FEP patients before the antipsychotic treatment. Multiple regression analysis demonstrated significant correlations between FEP and EGF, IL-1β and smoking. Treatment with antipsychotic drugs resulted in a statistically significant amelioration of the symptoms of psychosis, but caused a significant increase in the body mass index (BMI) of patients. Levels of EGF, IL-2, VEGF, IL-6, IFN-γ, IL-4, IL-8 and IL-1α were significantly lower in treated FEP patients compared to premedication levels. According to the present study, EGF and IL-1β are markers of FEP. Antipsychotic drug treatment resulted in a significant clinical improvement of FEP patients and the suppression of positive symptoms was correlated with the decreased levels of EGF, IL-2 and IL-4. EGF was the strongest marker of FEP and treatment efficiency among the measured cytokines and growth factors.


Acute pancreatitis associated with different combination therapies in patients infected with human immunodeficiency virus.
To assess the risk of acute pancreatitis in patients receiving various combinations of protease inhibitors, nucleoside reverse transcriptase inhibitors (NRTIs), and nonnucleoside reverse transcriptase inhibitors (NNRTIs) for treatment of human immunodeficiency virus (HIV) infection. Retrospective cohort study. Ohio Medicaid claims database, January 1997-December 2002. Four thousand nine hundred seventy-two patients with HIV infection who had received at least one antiretroviral drug. Three combination regimens were evaluated: didanosine plus other antiretroviral agents, protease inhibitors plus NRTIs or NNRTIs, and NRTI combinations (no didanosine) or NRTIs plus NNRTIs. We used Cox proportional hazard regression and Kaplan-Meier plots to examine the risk for acute pancreatitis. We identified 159 (3.2%) cases of acute pancreatitis during the study period. For patients who were newly treated for HIV, the incidence of acute pancreatitis was 1.95/100 person-years. Half of these cases developed within 500 days of the start of drug therapy. Hazard ratios (HRs) for acute pancreatitis were 39-54% higher for nonwhite patients than Caucasians and 240-290% higher for symptomatic versus nonsymptomatic patients. Hazard ratios also were significantly associated with increased age, liver injuries (HR 2.94, 6.73), and cardiovascular diseases (HR 1.68, 2.36), respectively, for both newly treated and previously diagnosed patients with HIV. The risk for patients receiving either protease inhibitors plus an NRTI or an NNRTI, or NRTI plus NNRTI combinations was not significantly different from the risk associated with didanosine combination therapy (p>0.10). The risk of acute pancreatitis was significantly associated with age, race, symptomatic HIV infection, and liver and cardiovascular diseases. However, risk did not differ significantly among patients with different antiretroviral regimens. Our results can be used by the medical community to enhance patient safety and minimize costly adverse drug reactions among patients with HIV infection.


Efficient membrane targeting of the chick nicotinic acetylcholine receptor α-subunit in stable insect cell lines.
We have synthesised the α-subunit of the chick nicotinic acetylcholine receptor (nAChR) in stable, continuous insect (Spodoptera frugiperda) cell lines. A cDNA was integrated randomly into the insect cell genome under control of a baculovius immediate early gene promoter. Transformed cells were obtained by co-transfection of the insect cells with pIEK1.nAChRα, encoding the α-subunit cDNA, and pIEK1.neo, encoding the neomycin resistance gene. G-418-resistant clones were selected and expanded into continuous cell lines synthesising the chick nAChR α-subunit. Using fluorescence microscopy and ligand binding studies we were able to demonstrate efficient membrane targeting of the receptor subunit in the insect cell plasma membrane. Stable insect cell lines may thus have significant advantages over transient baculovirus vectors for the synthesis and characterisation of heterologous receptor proteins.


The obesity associated FTO gene polymorphism and the risk of preeclampsia in Iranian women: A case-control study.
Preeclampsia (PE) is one of the leading disorders in pregnant women with maternal and fetal complications. Obesity is considered an important risk factor for the development of PE. Genetic variations in fat mass and obesity associated (FTO) gene may play a role in the development of PE. This study aimed to investigate the possible association between FTO gene rs9939609 and PE risk in a sample of Iranian pregnant women. In this case-control study, 312 pregnant women were included, including 128 with PE and 184 without PE. Demographic data and blood samples were obtained from all individuals. The genotyping of rs9939609 polymorphisms was performed by the tetra-primer amplification refractory mutation system-polymerase chain reaction (TP-ARMS-PCR) method, and the results of TP-ARMS-PCR were confirmed using DNA sequencing. The genotype frequency was 50%, 47.7%, and 2.3% in pregnant patients and 37%, 47.8%, and 15.2% in healthy controls for TT, AT, and AA, respectively. The risk of PE was significantly reduced in the pregnant women having the AA genotype. Based on the results of the present study, rs9939609 polymorphism in the FTO gene may play a protective role against PE. However, further studies are warranted. [Figure: see text].


Structure-activity relationships in aminosterol antibiotics: the effect of stereochemistry at the 7-OH group.
Squalamine and three aminosterol analogs have been shown to inhibit bacterial cell growth and induce lysis of large unilamellar phospholipid vesicles. The analogs differ in the identity of the polyamine attached at C3 of the sterol, and the stereochemistry of a hydroxyl substituent at C7. Analogs with a tetraammonium spermine polyamine are somewhat more active than analogs with a shorter trisammonium spermidine polyamine, and analogs with an axial (α) hydroxyl substituent at C7 are more active than analogs with the corresponding equatorial (β) hydroxyl group. There is some variability noted; the 7β-OH spermine analog is the most active compound against Escherichia coli, but the least effective against Pseudomonas aeruginosa. Lytic activity correlates well with antimicrobial activity of the compounds, but the lytic activity varies with the phospholipid composition of the vesicles.