Identification of molecular markers to follow up the bioremediation of sites contaminated with chlorinated compounds.
The use of microorganisms to clean up xenobiotics from polluted ecosystems (soil and water) represents an ecosustainable and powerful alternative to traditional remediation processes. Recent developments in molecular-biology-based techniques have led to rapid and sensitive strategies for monitoring and identifying bacteria and catabolic genes involved in the degradation of xenobiotics. This chapter provides a description of recently developed molecular-biology-based techniques, such as PCR with degenerate primers set, real-time quantitative PCR (qPCR), reverse transcription PCR (RT-PCR), southern blot hybridization, and long-range PCR, used to give a picture of the catabolically relevant microorganisms and of the functional genes present in a polluted system. By using a case study of a groundwater aquifer contaminated with 1,2-dichloroethane (1,2-DCA), we describe the identification of microorganisms potentially involved in the 1,2-DCA dehalorespiration (Dehalobacter sp. and Desulfitobacterium sp.) and a complete new gene cluster encoding for a 1,2-DCA reductive dehalogenase. The application of these techniques to bioremediation can improve our understanding of the inner mechanisms to evaluate the feasibility of a given treatment and provide us with a method to follow up bacteria and catabolic genes involved in the degradation of contaminants during the activities in situ.


Magnetic moment quantifications of small spherical objects in MRI.
The purpose of this work is to develop a method for accurately quantifying effective magnetic moments of spherical-like small objects from magnetic resonance imaging (MRI). A standard 3D gradient echo sequence with only one echo time is intended for our approach to measure the effective magnetic moment of a given object of interest. Our method sums over complex MR signals around the object and equates those sums to equations derived from the magnetostatic theory. With those equations, our method is able to determine the center of the object with subpixel precision. By rewriting those equations, the effective magnetic moment of the object becomes the only unknown to be solved. Each quantified effective magnetic moment has an uncertainty that is derived from the error propagation method. If the volume of the object can be measured from spin echo images, the susceptibility difference between the object and its surrounding can be further quantified from the effective magnetic moment. Numerical simulations, a variety of glass beads in phantom studies with different MR imaging parameters from a 1.5T machine, and measurements from a SQUID (superconducting quantum interference device) based magnetometer have been conducted to test the robustness of our method. Quantified effective magnetic moments and susceptibility differences from different imaging parameters and methods all agree with each other within two standard deviations of estimated uncertainties. An MRI method is developed to accurately quantify the effective magnetic moment of a given small object of interest. Most results are accurate within 10% of true values, and roughly half of the total results are accurate within 5% of true values using very reasonable imaging parameters. Our method is minimally affected by the partial volume, dephasing, and phase aliasing effects. Our next goal is to apply this method to in vivo studies.


The strategic use of biopsy in the diagnosis of coeliac disease in adults.
The paediatric guidelines support the use of the 'No Biopsy Approach' in the diagnosis of coeliac disease (CD). We aimed to determine the correlation between anti tissue transglutaminase (anti-TTG serology) ≥10 times the upper limit of normal (ULN), using the Celikey ® ELiA assay and histological findings. Our secondary aim was to determine the safety of this approach in our centre. A retrospective analysis of adult patients referred to a tertiary referral centre with raised anti-TTG titres and/or histological changes of coeliac on D2 biopsies between 2014 - 2019. Excluded patients were those who did not have a biopsy performed, or whose biopsy was unavailable for review, selective IgA deficiency, and gluten elimination prior to biopsy. Biopsies were classified according to Marsh, by two independent pathologists, blinded to the anti-TTG titre. 164 patients had positive anti-TTG serology and duodenal biopsy in our centre prior to starting a gluten free diet (GFD) in the period 2014 - 2019. Of these 164 patients (median age 40yrs, 62% female), 68 (33%) had an anti-TTG titre ≥10 x ULN, 99% of which had a Marsh grading ≥ 3 and 1% had a Marsh of 2 on biopsy. 91% had either a normal index gastroscopy or findings of mild gastritis/oesophagitis. We found a 98.5% positive predictive value (PPV) of determining CD (i.e., Marsh ≥ 3) in those with an anti-TTG ≥10 x ULN. In those with moderate to high-risk clinical suspicion of CD we propose that duodenal biopsy is unnecessary for diagnosis.


N-methyl-D-aspartic acid (NMDA) and non-NMDA receptors regulating hippocampal norepinephrine release. I. Location on axon terminals and pharmacological characterization.
The effects of endogenous and exogenous agonists at excitatory amino acid receptors mediating enhancement of [3H]norepinephrine [( 3H]NE) release have been investigated using superfused rat hippocampal synaptosomes. In Mg(++)-free medium L-glutamic acid (L-Glu), L-aspartic acid (L-Asp), N-methyl-D-aspartic acid (NMDA), kainic acid, (RS)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and quisqualic acid (QA) all increased the release of [3H]NE. L-Glu produced the largest effect. In the presence of Mg++ (1.2 mM), the effect of L-Glu decreased by about 40%; L-Asp and NMDA lost completely their activity while the effects of kainic acid, QA and AMPA did not change significantly. Similarly to NMDA, the effect of L-Asp was augmented by glycine and blocked by NMDA receptor antagonists, while it was insensitive to the non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). The effect of L-Glu on [3H] NE release was partly decreased by the NMDA receptor channel blocker (+)-5-methyl-10,11-dihydro-5-H-dibenzo[a,d]cycloepten-5,10-imine (MK-801) and partly by CNQX; when present together, the two antagonists completely abolished the L-Glu effect. The QA enhancement of [3H]NE release was antagonized by CNQX but it was insensitive to other classical non-NMDA receptor antagonists. 1) release-enhancing NMDA and non-NMDA receptors exist on noradrenergic axon terminals of rat hippocampus; 2) L-Asp appears to be a potent selective NMDA receptor agonist while L-Glu can activate also non-NMDA receptors; 3) the NE-releasing receptor activated by QA may represent a QA/AMPA receptor subtype.


Alcohol and alcohol problems research. 17. Malta.
This article is an enquiry into the current status of alcohol in Maltese culture. The responses of society to alcoholism depend on the way members of the community perceive the problems incurred by the use and abuse of a dependence producing substance like alcohol. These perceptions and subsequent responses are very much influenced by prevailing attitudes and beliefs. Malta is a melting point of cultures. This factor, together with a high density population and Malta's geopolitical strategic position, combine to make Malta a tolerant society. There is a laissez-faire response to alcoholism, at least partly due to the present inability to identify the need to take appropriate measures. The police force, medical profession and politicians still do not feel the responsibility or the need to provide effective laws and regulations, specialized treatment services or educative programmes on alcohol-related issues. A systematic enquiry is needed urgently to determine the severity and degree of the problems posed by alcohol abuse among the Maltese. Such an enquiry should be followed by a well planned national policy which includes local approaches and interventions. Finally, these interventions must be evaluated frequently and developed to achieve better results in the future.


Genome-wide association study in individuals of European and African ancestry and multi-trait analysis of opioid use disorder identifies 19 independent genome-wide significant risk loci.
Despite the large toll of opioid use disorder (OUD), genome-wide association studies (GWAS) of OUD to date have yielded few susceptibility loci. We performed a large-scale GWAS of OUD in individuals of European (EUR) and African (AFR) ancestry, optimizing genetic informativeness by performing MTAG (Multi-trait analysis of GWAS) with genetically correlated substance use disorders (SUDs). Meta-analysis included seven cohorts: the Million Veteran Program, Psychiatric Genomics Consortium, iPSYCH, FinnGen, Partners Biobank, BioVU, and Yale-Penn 3, resulting in a total N = 639,063 (Ncases = 20,686;Neffective = 77,026) across ancestries. OUD cases were defined as having a lifetime OUD diagnosis, and controls as anyone not known to meet OUD criteria. We estimated SNP-heritability (h2SNP) and genetic correlations (rg). Based on genetic correlation, we performed MTAG on OUD, alcohol use disorder (AUD), and cannabis use disorder (CanUD). A leave-one-out polygenic risk score (PRS) analysis was performed to compare OUD and OUD-MTAG PRS as predictors of OUD case status in Yale-Penn 3. The EUR meta-analysis identified three genome-wide significant (GWS; p ≤ 5 × 10-8) lead SNPs-one at FURIN (rs11372849; p = 9.54 × 10-10) and two OPRM1 variants (rs1799971, p = 4.92 × 10-09; rs79704991, p = 1.11 × 10-08; r2 = 0.02). Rs1799971 (p = 4.91 × 10-08) and another OPRM1 variant (rs9478500; p = 1.95 × 10-08; r2 = 0.03) were identified in the cross-ancestry meta-analysis. Estimated h2SNP was 12.75%, with strong rg with CanUD (rg = 0.82; p = 1.14 × 10-47) and AUD (rg = 0.77; p = 6.36 × 10-78). The OUD-MTAG resulted in a GWAS Nequivalent = 128,748 and 18 independent GWS loci, some mapping to genes or gene regions that have previously been associated with psychiatric or addiction phenotypes. The OUD-MTAG PRS accounted for 3.81% of OUD variance (beta = 0.61;s.e. = 0.066; p = 2.00 × 10-16) compared to 2.41% (beta = 0.45; s.e. = 0.058; p = 2.90 × 10-13) explained by the OUD PRS. The current study identified OUD variant associations at OPRM1, single variant associations with FURIN, and 18 GWS associations in the OUD-MTAG. The genetic architecture of OUD is likely influenced by both OUD-specific loci and loci shared across SUDs.


Socioeconomic inequalities in health-related quality of life between men and women, 5 years after a coronary angiography.
The aim of this study is to measure gender differences in health-related quality of life (HRQOL) among men and women patients with cardiovascular diseases (CVD), and to assess the impact of socioeconomic factors on HRQOL between men and women, 5 years after a coronary angiography. The study included 1,289 out of 4,391 patients who had undergone an angiography in the National Institute for Cardiac Surgery and Interventional Cardiology, Luxembourg in 2008/2009. Four indicators of the WHOQOL-BREF questionnaire (Self-rated health, Quality of life, Physical health, and Psychological health) were used in this study as interest variables. To assess the socioeconomic inequalities in HRQOL between men and women, general linear models were constructed for every indicator, with educational level and living conditions as predictors, and demographic variables, cardiovascular risk factors, and cardiovascular events as covariates. Women were older than men (71.5 versus 68.1, p <0.0001) and less likely to be married. HRQOL was significantly different between men and women despite the fact they had the same socioeconomic status. The average score for overall health was 3.7/5 for men versus 3.5/5 for women; similarly, the life quality score was 3.8/5 for men versus 3.6/5 for women. Education level and living conditions were associated with lower HRQOL scores in men and women. The findings showed that women have lower HRQOL than men regarding self-rated health, quality of life, and the WHOQOL-BREF physical and psychological domains 5 years after a coronary angiography. Socioeconomic inequalities affect HRQOL, and their influence was similar in both men and women. Socioeconomic inequalities in HRQOL in women and men with CVD are strong 5 years after a coronary angiography. Taking into account differences in gender and socioeconomic status in intervention strategies to substantially reduce the differences observed between women and men could help improve the effectiveness of secondary prevention.


Perceived Relative Deprivation Across the Adult Lifespan: An Examination of Aging and Cohort Effects.
Despite being a core psychological construct for over 70 years, research has yet to examine how perceptions of deprivation relative to other individuals and/or groups develop across adulthood. As such, this preregistered study uses cohort-sequential latent growth modeling to examine changes in individual- and group-based relative deprivation (IRD and GRD, respectively) across the adult lifespan. Across 10 annual assessments of a nationwide random sample of adults (Ntotal = 58,878; ethnic minority n = 11,927; 62.7% women; ages 21-80), mean levels of IRD trended downward across the lifespan, whereas mean levels of GRD generally increased from young-to-middle adulthood before declining across late adulthood. Subtle cohort effects emerged for both constructs, although both IRD and GRD largely followed a normative aging process. Critically, the development of GRD-but not IRD-differed between ethnic groups, providing insights into how one's objective status may shape subjective (dis)advantage over time.


Vascular endothelial growth factor-A signaling in bone marrow-derived endothelial progenitor cells exposed to hypoxic stress.
Bone marrow-derived endothelial progenitor cells (BM-EPCs) are stimulated by vascular endothelial growth factor-A (VEGF-A) and other potent proangiogenic factors. During angiogenesis, an increase in VEGF-A expression stimulates BM-EPCs to enhance endothelial tube formation and contribute to an increase in microvessel density. Hypoxia is known to produce an enhanced angiogenic response and heightened levels of VEGF-A have been seen in oxygen deprived epithelial and endothelial cells, yet the pathways for VEGF-A signaling in BM-EPCs have not been described. This study explores the influence of hypoxia on VEGF-A signaling in rat BM-EPCs utilizing a novel proteomic strategy to directly identify interacting downstream components of the combined VEGF receptor(s) signaling pathways, gene expression analysis, and functional phenotyping. VEGF-A signaling network analysis following liquid chromatographic separation and tandem mass spectrometry revealed proteins related to inositol/calcium signaling, nitric oxide signaling, cell survival, cell migration, and inflammatory responses. Alterations in BM-EPC expression of common angiogenic genes and tube formation in response to VEGF-A during hypoxia were measured and combined with the proteomic analysis to enhance and support the signaling pathways detected. BM-EPC tube formation assays in response to VEGF-A exhibited little tube formation; however, a cell projection/migratory phenotype supported the signaling data. Additionally, a novel assay measuring BM-EPC incorporation into preformed endothelial cell tubes indicated a significant increase of incorporated BM-EPCs after pretreatment with VEGF-A during hypoxia. This study verifies known VEGF-A pathway components and reveals several unidentified mechanisms of VEGF-A signaling in BM-EPCs during hypoxia that may be important for migration to sites of vascular regeneration.


New isoforms of Ca2+/calmodulin-dependent protein kinase II in smooth muscle.
Four novel isoforms of Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) were found in rat aorta smooth muscle. Two of them were related to gamma-isoform of brain CaM kinase II (gamma-a). Differences in the primary structure of these isoforms were located in the variable region. One of them (gamma-b) contained 23 unique amino acid residues, whereas the other (gamma-c) did not contain this sequence. Both isoforms lacked the two segments (Val-316 to Gln-337 and Lys-353 to Leu-362) present in gamma-a. The DNA sequence of these gamma-isoforms except the variable region was exactly the same, suggesting that they are produced by alternative splicing. Another two isoforms were related to the delta-isoform of brain CaM kinase II (delta-a). delta-b contained a unique 11-residue sequence in the variable region whereas delta-c did not. As found for gamma-isoforms, the sequence analysis suggested that the three delta-isoforms are also produced by alternative splicing. Analysis of RNA by reverse transcription PCR confirmed the existence of specific messages for gamma-b, delta-a and delta-b. The variety of isoforms of CaM kinase II suggest that each isoform may play a specialized role in cell regulation.


PK, PD, and interactions: the new scenario with JAK inhibitors and S1P receptor modulators, two classes of small molecule drugs, in IBD.
Inflammatory bowel diseases (IBDs) are immune-mediated chronic inflammatory disorders of the gastrointestinal tract whose pathogenesis is not yet fully understood. Despite the advent of biological agents, there are still unmet needs for IBD patients, due to suboptimal rate of sustained remission achieved. Small molecule drugs (SMDs), the next generation of selective drugs in IBD, show promising results in ongoing trials. We describe the pharmacodynamics and pharmacokinetic features of novel SMDs and their main differences with biologic agents. Small molecule drugs are a promising class of drugs for the treatment of ulcerative colitis and Crohn's disease with good results in inducing and maintaining remission. Hence, over the next few years physicians will have numerous options of small molecule drugs for the treatment of patients with IBD. This group of drugs are potentially easier to use over biological agents due to pharmacokinetic features such as oral administration, short half-life, high volume of distribution, and lack of immunogenicity. On the other hand, drug-drug interactions can happen with small-molecule drugs, principally due to competitive metabolic and clearance mechanisms.


Schizophrenia, hemodialysis, and the placebo effect. Results and issues.
This report presents the results of a double-blind study that examines the impact of hemodialysis on patients with chronic schizophrenia. Twelve patients were randomly assigned to sham and 12 patients to active dialysis. All patients experienced a drug washout period and were then followed up over an eight-week period. A statistically significant condition by weeks interaction was found, with patients in the sham group improving and patients in the active dialysis group decompensating over the last weeks of the trial. This complex finding fails to support the hypothesized efficacy of hemodialysis in the treatment of schizophrenia but also fails to support a simple placebo explanation for positive results reported in earlier open studies. The methodologic complications are reviewed, and a possible direction for future research is presented.


Free intestinal autografts for reconstruction following pharyngolaryngoesophagectomy.
After pharyngolaryngoesophagectomy in four patients, free intestinal autografts were used for reconstruction. A microvascular technique and a simultaneous two team approach allowed this to be done immediately following the resection. No graft failures have occurred. Good function was present in three of the four patients at the last follow-up examination or at death. Follow-up intervals vary from seven to 15 months postoperatively. Special problems, such as irradiation, tumor site and the relative merits of the large versus the small intestine, are discussed.


Smile aesthetics.
The aim of this article is to outline some of the factors practitioners should take into consideration when planning optimal smile aesthetics. The components of the smile that will be discussed include the smile arc, incisor/gingival display, smile width, gingival aesthetics, tooth proportionality/symmetry, contacts/connectors/embrasures and the dental midlines. A systematic evaluation of the components of the smile when planning treatment will help to improve smile aesthetics and produce greater patient and dentist satisfaction.


[Tapia's syndrome following transesophageal echocardiography during an open-heart operation: a case report].
A 67-year-old man presented with hoarseness, dysarthria and deviation of the tongue to the left side the day after the open-heart operation under general anesthesia. Brain MRI demonstrated no causal lesion, and laryngoscope showed left vocal cord abductor palsy, so we diagnosed him with Tapia's syndrome (i.e., concomitant paralysis of the left recurrent and hypoglossal nerve). His neurological symptoms recovered gradually and improved completely four months after the onset. Tapia's syndrome is a rare condition caused by the extra cranial lesion of the recurrent laryngeal branch of the vagus nerve and the hypoglossal nerve, and mostly described as a complication of tracheal intubation. In this case, transesophageal echo probe has been held in the left side of the pharynx, so compression to the posterior wall of pharynx by the probe resulted in this condition, and to the best of our knowledge, this is the first report of Tapia's syndrome due to transesophageal echocardiography during an open-heart operation. This rare syndrome should be considered as a differential diagnosis of dysarthria and tongue deviation after a procedure associated with compression to the pharynx.


Qualitative evidence regarding the experience of receiving and providing care for mental health conditions in non-specialist settings in low-income and middle-income countries: a systematic review of reviews.
This review of reviews synthesises qualitative evidence on the experiences of receiving and providing care and treatment for mental health conditions in non-specialist settings in low-income and middle-income countries (LMICs), and the factors that influence the provision and uptake of such services. Database searches were conducted in PubMed/MEDLINE, EMBASE, PsycINFO, CINAHL, Scopus, African Index Medicus and Global Index Medicus, supplemented by screening repositories of systematic reviews protocols and contacting authors. The evidence synthesis drew on deductive and inductive approaches: a framework analysis approach was used for the initial coding structure, after which the results synthesis was refined further through reviewing and regrouping the initial coding through thematic synthesis principles. Nine reviews met inclusion criteria and reported on a range of factors related to the provision and uptake of mental healthcare by non-specialist health workers in LMICs: (1) health worker competency, (2) availability of resources, (3) recipient-related and provider-related characteristics, (4) service accessibility, (5) sociocultural acceptability and (6) vulnerable groups for whom barrier to care were potentially exacerbated. This review provides nuanced and contextualised insights regarding the experiences of receiving and providing care for mental health conditions in LMICs, including barriers influencing service provision and uptake. It is important to ensure mental healthcare in non-specialist settings in LMICs is delivered in a manner which is feasible, acceptable and culturally appropriate in order to improve access to care, reducing stigma and promoting better overall health and well-being for individuals and communities.


Hydrogen-rich saline protects intestinal epithelial tight junction barrier in rats with intestinal ischemia-reperfusion injury by inhibiting endoplasmic reticulum stress-induced apoptosis pathway.
To investigate the effects of hydrogen-rich saline (HRS) on intestinal epithelial tight junction (TJ) barrier in rats with intestinal ischemia-reperfusion injury (IIRI). Thirty-two healthy male Sprague-Dawley (SD) rats were randomly divided into four groups (n = 8 each): Sham group, I/R group, HRS group and 4-PBA group. After 45 min of ischemia and 6 h of reperfusion, the rats were sacrificed to collect serum and ileum for detection. Hematoxylin and eosin (H&E) staining was used to observe the morphology of small intestine. The serum expression levels of intestinal fatty acid binding protein (IFABP), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) were determined by enzyme linked immunosorbent assay (ELISA). Imunohistochemistry, immunofluorescence and Western blot were used to detect key proteins in intestinal epithelial TJs, ERS, and ERS-induced apoptosis, including occludin, zonula occludens-1 (ZO-1), glucose-regulated protein 78 (GRP78), X-box binding protein-1 (XBP1), C/EBP-homologous protein (CHOP) and caspase-3. Data was presented as mean ± SEM and compared using one-way ANOVA. A p-value <0.05 was considered significant. Compared with rats in the I/R group, the Chiu score of ileum damage in the HRS group and 4-PBA group were lower. The levels of serum IFABP, TNF-α, and IL-1β were statistically significant among the groups. Increased expression of TJ proteins occludin and ZO-1 by reducing various parameters of ERS and ERS-induced apoptosis evidenced by down-regulation of the protein levels of GRP78, XBP1, CHOP and caspase-3 were shown in the HRS and 4-PBA groups. HRS had potential protective effects on intestinal barrier in IIRI rats. This study suggested that inhibition of excessive ERS and ERS-induced apoptosis by HRS may reduce intestinal epithelial cells damage and maintain the integrity of intestinal epithelial TJ barrier in rats with IIRI.


Morphometry of the orbital region soft tissues in Down syndrome.
The orbital region of subjects with Down syndrome (DS) has been scanty described so far. We wanted to detail the morphologic characteristics of the soft tissue orbital region in Italian and North Sudanese subjects with DS. The three-dimensional coordinates of 10 landmarks on the orbital soft tissues were obtained using computerized anthropometry in 53 Italian and 64 North Sudanese subjects with DS aged 4 to 52 years, and in 461 (Italian) and 682 (North Sudanese) sex- and age-matched controls. From the landmarks, linear distances, ratios, areas, and angles were calculated, z scores computed, and compared by t-tests and analyses of covariance. In North Sudanese DS subjects, intercanthal width and height-to-length ratio were increased; biorbital width, eye height, length, and area were reduced. Eye fissure and orbital inclinations relative to Frankfort plane were reduced, whereas orbital inclinations versus the true horizontal were increased. In Italian DS men, orbital height and height-to-length ratio were increased, eye length was decreased; orbital inclination versus the true horizontal was increased. For almost all measurements, a significant effect of age was found. No effects of sex were found. Ethnic group influenced orbital height, area, and orbital inclination versus Frankfort plane. All paired measurements had similar discrepancies on both sides. The orbital soft tissues of North Sudanese DS subjects differed from those of their reference subjects, but this was only partially true for Italian subjects. The 2 ethnic groups had different alterations in their soft tissue orbital regions that were influenced by age, but not by sex.


Factors influencing local recurrence after excision and radiotherapy for primary breast cancer.
Between November 1979 and December 1986, 263 patients were treated for primary breast cancer by local excision and radiotherapy at the City Hospital, Nottingham. Local recurrence within the treated breast occurred in 56 patients (21 per cent), in 18 (6.8 per cent) of whom it was gross and uncontrollable. An analysis of clinicopathological features shows patient age, nodal status, tumour size, presence of definitive vascular invasion, adjacent ductal carcinoma in situ and grade to be predictive of local recurrence. A Cox's multivariate analysis of these factors shows the first four to be independently significant. The factors can be combined as a prognostic index which allows identification of patients at high risk of local recurrence. On the basis of these findings we have altered our selection policy for patients suitable for breast conservation.


Erythrocytic glucose-6-phosphate dehydrogenase measured by a differential pH technique.
We propose a new quantitative electrochemical method for determining glucose-6-phosphate dehydrogenase (EC 18.104.22.168) activity in purified erythrocytes or in whole blood, based on measurement of the pH change caused by oxidation of glucose 6-phosphate to 6-phosphogluconic acid, with simultaneous reduction of NADP+ to NADPH + H+. No sample pretreatment (e.g., preparation of hemolysate) is needed, and the automatic correction for sample blanks obviates interference from 6-phosphogluconate dehydrogenase (EC 22.214.171.124). The method is simple and fast, and the standard curve is linear to at least 2200 U/L at 37 degrees C. The within-day CV was 3.9% for activities in healthy individuals (mean value 1204 U/L), and 10% for deficient ones (classified as belonging to class II, mean value 407 U/L). Results (y) correlated well with those obtained with the WHO-recommended method (x): y = 1.13x + 0.02 (r = 0.971) for purified erythrocytes. Normal reference intervals are reported for the enzyme in purified erythrocytes and in whole blood.


Immediate CEA for symptomatic carotid disease preferably performed under local anaesthesia is safe.
Previous general reservations against carotid endarterectomy (CEA) early after stroke, which were primarily based on concerns of postoperative intracerebral hemorrhage, are resolved. Moreover, a delay of surgery is proofed to be associated with a risk of recurrent cerebral ischemia. However, the complication rate of CEA seems to increase with less time interval to the onset of symptoms. The main purpose of this study was to assess the safety of very early CEA. Patients having a symptomatic high-grade (> 70%) internal carotid artery (ICA) stenosis were referred by neurologists for CEA within different timeframes, so that they were later differentiated depending on whether surgery was performed within 2 days (immediate CEA = iCEA) or 2 weeks (urgent CEA = uCEA) after neurological deficits have occurred primarily. The perioperative complication rate in these groups was than evaluated and compared. From January 2000 until August 2006 130 consecutive patients (median age 68 years, range: 42-90; 66% male, 34%female)presenting with an ipsilateral TIA (n = 80), stroke (n = 50) underwent iCEA (n = 40) or uCEA (n = 90). Demographic and clinical characteristics were equally distributed between treatment groups. Mostly (121/130), CEA was performed under local anaesthesia with selective shunt use which became necessary in 26%. Besides postoperative hemorrhage (n = 4), cardiac complications (n = 2) and temporary cranial nerve lesions (n = 2), new perioperative neurological deficits occurred in total in 8 patients of which 6 were temporary. The other 2 patients developed strokes of which one patient died. Therefore, the combined stroke- and mortality rate was 1.5% (2/130) for the whole study population. With regard to the timing of surgery, a single incident was observed after iCEA (1/40) which also was the only intracerebral hemorrhage. It seems that patients with a symptomatic high-grade ICA stenosis can undergo CEA particularly under local anaesthesia as soon as possible without anticipating an increased complication rate.


Evaluating pooled testing for asymptomatic screening of healthcare workers in hospitals.
There is evidence that during the COVID pandemic, a number of patient and HCW infections were nosocomial. Various measures were put in place to try to reduce these infections including developing asymptomatic PCR (polymerase chain reaction) testing schemes for healthcare workers. Regularly testing all healthcare workers requires many tests while reducing this number by only testing some healthcare workers can result in undetected cases. An efficient way to test as many individuals as possible with a limited testing capacity is to consider pooling multiple samples to be analysed with a single test (known as pooled testing). Two different pooled testing schemes for the asymptomatic testing are evaluated using an individual-based model representing the transmission of SARS-CoV-2 in a 'typical' English hospital. We adapt the modelling to reflect two scenarios: a) a retrospective look at earlier SARS-CoV-2 variants under lockdown or social restrictions, and b) transitioning back to 'normal life' without lockdown and with the omicron variant. The two pooled testing schemes analysed differ in the population that is eligible for testing. In the 'ward' testing scheme only healthcare workers who work on a single ward are eligible and in the 'full' testing scheme all healthcare workers are eligible including those that move across wards. Both pooled schemes are compared against the baseline scheme which tests only symptomatic healthcare workers. Including a pooled asymptomatic testing scheme is found to have a modest (albeit statistically significant) effect, reducing the total number of nosocomial healthcare worker infections by about 2[Formula: see text] in both the lockdown and non-lockdown setting. However, this reduction must be balanced with the increase in cost and healthcare worker isolations. Both ward and full testing reduce HCW infections similarly but the cost for ward testing is much less. We also consider the use of lateral flow devices (LFDs) for follow-up testing. Considering LFDs reduces cost and time but LFDs have a different error profile to PCR tests. Whether a PCR-only or PCR and LFD ward testing scheme is chosen depends on the metrics of most interest to policy makers, the virus prevalence and whether there is a lockdown.


Assessment of Body Composition in Oncologic Patients: Experimental Survey on the Role of Bioimpedentiometric Analysis.
Overall survival of oncologic patients is strongly influenced by the incidence of malnutrition, with subsequent loss of muscle mass until sarcopenia. In this respect, the assessment of body composition has a pivotal role in order to manage the clinical consequences of muscle loss. This study focuses on the body composition assessment in oncologic patients, following a diet plan in order to detect and contrast neoplastic cachexia. 35 oncologic patients were enrolled and divided into two groups: 24 responders (R) and 11 non-responders (NR). Anthropometric data were collected and body composition was assessed through bioimpedentiometry. All patients received an individualized normocaloric diet. Energetic content was assigned on the basis of individual basal metabolic rate estimated by BIA. All patients were revaluated by anthropometry and bioimpedentiometry 3 and 6 months thereafter. Comparing the two male groups R and NR after 6 months, the former maintained almost the same confidence interval, unlike the latter whose interval increased, indicating a light worsening both of body composition and of clinical conditions. Furthermore, in the same male responder group, an improvement of the phase angle (PhA), a positive prognostic factor, was found, with an average weight loss between 2,6 - 3 kg of fat mass (FM). Besides, data show how the R female group maintained the confidence interval unlike the NR female group, whose confidence interval remarkably increased, therefore indicating an important alteration of body composition and subsequently a clinical conditions worsening. Finally, comparing the NR male group with the NR female group after 6 months, a remarkable worsening of body composition (marker of tissue decay and damage) could be noticed in the latter group. After 6 months, NR patients show worse results (both in the body composition and in the clinical conditions) compared to the responders, with particularly worse results in females, probably because of a genetically determined sex-related smaller muscle cells and inferior muscular strength. A multidimensional assessment of oncologic patient is necessary, with a special attention to nutritional evaluation and body composition monitoring, in order to avoid malnutrition and subsequent further clinical worsening.


Translation of the hepatitis B virus P gene by ribosomal scanning as an alternative to internal initiation.
The hepatitis B virus (HBV) P gene which encodes the reverse transcriptase and other proteins required for replication is expressed on the bicistronic mRNA pregenome which also encodes the capsid protein in its first cistron. Recent results have suggested that the hepadnaviral P gene is translated by internal entry of ribosomes upstream from the P gene, in the overlapping C gene. Using a reporter gene fused to the HBV C or P gene, we demonstrate that the C sequence does not allow internal initiation of translation. Alternatively, our results support a model in which the HBV P gene is translated by ribosomes which scan from the capped extremity of the bicistronic mRNA pregenome. The mechanism by which the ribosomes scan past four AUGs before they initiate translation at the P AUG was analyzed. Our results show that these AUGs are skipped via two mechanisms: leaky scanning on AUGs in a weak or suboptimal initiation context and translation of an out-of-C-frame minicistron followed by reinitiation at P AUG. The minicistron translation allows ribosomes to bypass an AUG in a favorable context that would otherwise be used as a start codon for translation of a truncated capsid protein. Our results suggest that this elaborated scanning mechanism permits the coordinate expression of the HBV C and P genes on the viral bicistronic mRNA pregenome.


Angiotensin II receptor blockers and cardiovascular protection: focus on left ventricular hypertrophy regression and atrial fibrillation prevention.
Left ventricular hypertrophy (LVH) and atrial fibrillation (AF) are strong predictors of cardiovascular (CV) morbidity and mortality, independently of blood pressure levels and other modifiable and nonmodifiable risk factors. The actions of circulating and tissue angiotensin II, mediated by AT1 receptors, play an important role in the development of a wide spectrum of cardiovascular alterations, including LVH, atrial enlargement and AF. Growing experimental and clinical evidence suggests that antihypertensive drugs may exert different effects on LVH regression and new onset AF in the setting of arterial hypertension. Since a number of large and adequately designed studies have found angiotensin II receptor blockers (ARBs) to be more effective in reducing LVH than beta-blockers and data are also available showing their effectiveness in preventing new or recurrent AF, it is reasonable to consider this class of drugs among first line therapies in patients with hypertension and LVH (a very high risk phenotype predisposing to AF) and as adjunctive therapy to antiarrhythmic agents in patients undergoing pharmacological or electrical cardioversion of AF.


The relationship between obesity and injuries among U.S. adults.
To quantify the relationship between body mass index (BMI) and rates of medically attended injuries by mechanism (overall, fall, motor vehicle, and sport-related) and by nature (strain/sprain, lower extremity fracture, and dislocations), and between BMI and injury treatment costs. Cross-sectional analysis. SETTING. The noninstitutionalized population of the United States. The 1999-2000, 2000-2001, and 2001-2002 waves of the Medical Expenditure Panel Survey, a large, nationally representative dataset, were combined to create the analysis sample. The final sample included 42,304 adults. MEASURES. Medically attended injury rates by mechanism and nature of injury and related treatment costs. Logistic regressions were used to separately estimate the odds of sustaining any injury by mechanism or by nature for overweight (25 < BMI : 29.9) and three categories of obese individuals compared with those who were normal weight. A second set of regressions tested whether, given that an injury occurred, obese individuals had greater injury treatment costs. RESULTS. Slightly more than one in five adults sustain an injury each year that requires medical treatment. The odds of sustaining an injury are 15% (overweight) to 48% (Class III obesity) greater among those with excess weight. Conditional on sustaining an injury, BMI did not have a significant impact on injury treatment costs. Our findings show a clear association between BMI and the probability of sustaining an injury. If increasing BMI is causing the rise in injury rates, then the incidence of injuries, including those related to falls, sprains/strains, lower extremity fractures, and joint dislocations, are likely to increase as the prevalence of obesity increases.


NKT cells inhibit antigen-specific effector CD8 T cell induction to skin viral proteins.
We recently demonstrated that CD1d-restricted NKT cells resident in skin can inhibit CD8 T cell-mediated graft rejection of human papillomavirus E7-expressing skin through an IFN-γ-dependent mechanism. In this study, we examined the role of systemically derived NKT cells in regulating the rejection of skin grafts expressing viral proteins. In lymph nodes draining transplanted skin, Ag-specific CD8 T cell proliferation, cytokine production, and cytotoxic activity were impaired by NKT cells. NKT cell suppression was mediated via CD11c(+) dendritic cells. Inhibition of CD8 T cell function did not require Foxp3(+) regulatory T cells or NKT cell-secreted IFN-γ, IL-10, or IL-17. Thus, following skin grafting or immunization with human papillomavirus-E7 oncoprotein, NKT cells reduce the capacity of draining lymph node-resident APCs to cross-present Ag to CD8 T cell precursors, as evidenced by impaired expansion and differentiation to Ag-specific CD8 T effector cells. Therefore, in the context of viral Ag challenge in the skin, systemic NKT cells limit the capacity for effective priming of adaptive immunity.


Genetic improvement of poplar feedstock quality for ethanol production.
Opportunities for matching chemical and physical properties of woody feedstocks to ethanol production process requirements via genetic improvement have long been recognized. Exploitation is now feasible owing to advances in trait measurement, breeding, and gene transfer technologies. Poplar genetic parameters are favorable largely for reducing lignin and increasing cellulose contents and specific gravity. Transgenic poplars with decreased lignin and increased cellulose contents, but otherwise normal growth and development, have been produced via genetic transformation. The long-standing debate on feasibility has thus become one of when, not if, designer varieties will become available.


Fragile X Premutation: Medications, Therapy and Lifestyle Advice.
The fragile X premutation is characterized by 55-200 CGG repeats in the 5' untranslated region of FMR1, whereas full fragile X mutation has greater than 200 repeats and full methylation, which manifests as fragile X syndrome (FXS). The premutation spectrum of clinical involvement includes fragile X-associated tremor/ataxia syndrome (FXTAS), fragile X-associated primary ovarian insufficiency (FXPOI), and fragile X-associated neuropsychiatric disorders (FXAND). In addition, premutation carriers also suffer from various other health problems such as endocrine abnormalities and autoimmune problems. In this paper, we have discussed different health issues faced by the carriers and interventions including medications, therapy and lifestyle changes that could improve their health.


Thioester and Oxyester Linkages in the Ubiquitin System.
The traditional textbook describes ubiquitylation as the conjugation of ubiquitin to a target by forming a covalent bond connecting ubiquitin's carboxy-terminal glycine residue with an acceptor amino acid like lysine or amino-terminal methionine in the substrate protein. While this adequately depicts a significant fraction of cellular ubiquitylation processes, a growing number of ubiquitin modifications do not follow this rule. Recent data demonstrate that ubiquitin can also be efficiently attached to other amino acids, such as cysteine, serine, and threonine, via ester bonding. Initially observed for a virus-encoded ubiquitin ligase, which targets a cysteine residue in a host protein to initiate its degradation, ester-linked ubiquitylation is now shown to also drive regular cellular processes. These ubiquitylation events expand the complexity and diversity of ubiquitin signaling and broaden the capability of cellular messages in the so-called ubiquitin code. Still, questions on the prevalence, relevance, and involvement in physiological and cellular functions await clearing. In this review, we aim to summarize our knowledge on ester-linked ubiquitylation and introduce experimental strategies to circumvent technical issues that complicate analysis of this uncommon posttranslational modification.


Current status of understanding of the genetic etiology of coronary heart disease.
Coronary heart disease (CHD), synonymously known as coronary artery disease (CAD) is the most predominant among the cardiovascular diseases and ranked number one in prevalence among the developing countries. CHD is a multifactorial disease involving both genetic and environmental factors and is primarily caused due to a process of progressive damage of coronary arteries called atherosclerosis. We present here a comprehensive review of molecular genetic studies conducted so far on CAD. The information was gathered through the internet using appropriate search terms for CHD/CAD. We also compiled the relevant information from the following websites: http://www.bioguo.org/CADgene/and http://www.genome.gov. Besides several Mendelian forms of the CHD, ~300 more genes have been identified in different studies through candidate gene approach. Additionally 32 more loci have been identified through genome wide association studies that include 9p21.3 as the most replicated genetic locus across the globe. Nevertheless, overall, these studies have been characterized by a relative lack of consistency in the association pattern across the populations. A fair degree of ethnic variation in the nature of association of different genetic variants with the disease has also been apparent. Pleiotropic effects of genes, existence of subclinical phenotypes and genetic heterogeneity appear to have been the limiting factors for developing a genetic risk profile test for the disease. Given the high prevalence of this disease in India, the presence of environmental triggers and genetic variation, it would be prudent to conduct multi-ethnic large-scale studies in India, representing the subcontinent as a whole-there have been a very limited number of molecular genetic studies on Indian populations.


Negative inotropic effect of extracellular calcium buffering in cardiac muscle.
Heart muscle contracts more vigorously when calcium levels are raised. A transient depletion of calcium from restricted extracellular spaces occurs with each contraction. We decided to maintain the concentration of this ion at a constant level by using an external calcium buffering system. It was found that buffering calcium at a millimolar level (using citrate as a buffer) caused a decrease, rather than an increase in the strength of contraction. The mean reduction in peak tension was by 27% in guinea pig and by 50.5% in frog atrium. This finding is analyzed; its most plausible explanation is the hypothesis that the buffer dissipates a calcium inhomogeneity, consisting of a higher calcium concentration adjacent to the membrane. Alternative interpretations such as intracellular acidosis, were tested experimentally and ruled out.


Unilateral lingual nerve transection alters jaw-tongue coordination during mastication in pigs.
During chewing, movements and deformations of the tongue are coordinated with jaw movements to manage and manipulate the bolus and avoid injury. Individuals with injuries to the lingual nerve report both tongue injuries due to biting and difficulties in chewing, primarily because of impaired bolus management, suggesting that jaw-tongue coordination relies on intact lingual afferents. Here, we investigate how unilateral lingual nerve (LN) transection affects jaw-tongue coordination in an animal model (pig, Sus scrofa). Temporal coordination between jaw pitch (opening-closing) and 1) anteroposterior tongue position (i.e., protraction-retraction), 2) anteroposterior tongue length, and 3) mediolateral tongue width was compared between pre- and post-LN transection using cross-correlation analyses. Overall, following LN transection, the lag between jaw pitch and the majority of tongue kinematics decreased significantly, demonstrating that sensory loss from the tongue alters jaw-tongue coordination. In addition, decrease in jaw-tongue lag suggests that, following LN transection, tongue movements and deformations occur earlier in the gape cycle than when the lingual sensory afferents are intact. If the velocity of tongue movements and deformations remains constant, earlier occurrence can reflect less pronounced movements, possibly to avoid injuries. The results of this study demonstrate that lingual afferents participate in chewing by assisting with coordinating the timing of jaw and tongue movements. The observed changes may affect bolus management performance and/or may represent protective strategies because of altered somatosensory awareness of the tongue.NEW & NOTEWORTHY Chewing requires coordination between tongue and jaw movements. We compared the coordination of tongue movements and deformation relative to jaw opening-closing movements pre- and post-lingual nerve transection during chewing in pigs. These experiments reveal that the timing of jaw-tongue coordination is altered following unilateral disruption of sensory information from the tongue. Therefore, maintenance of jaw-tongue coordination requires bilateral sensory information from the tongue.


Rapid Serological Tests Ineffectively Screen for HIV Exposure in HIV-Positive Infants.
Data on the performance and utility of rapid serological tests in infants to determine HIV exposure are unclear and in some instances contradictory. This study sought to understand the performance of rapid serological tests in high HIV burden, high Option B+ coverage settings to be used as an HIV exposure screening tool. A total of 3600 infants up to 24 months of age at 4 regional hospitals in Uganda were systematically enrolled and tested simultaneously using both HIV rapid serological and nucleic acid-based tests. Only 58 of the 94 HIV-positive infants who received both rapid serological and nucleic acid-based tests were positive with the rapid serological test (sensitivity: 61.7%; 95% confidence interval: 51.1 to 71.5). Using rapid serological tests to screen infants for exposure to HIV and follow-up nucleic acid-based testing would have missed 38.3% (36 of 94) of HIV-positive infants. Finally, several HIV-positive infants who were negative by rapid serological test presented to well-child entry points and were considered healthy. All 3 HIV-positive infants presenting to outreach and immunization were negative by rapid serological testing and 73% (8 of 11) presenting to outpatient. These data suggest that the use of rapid serological tests may have inadequate performance as an indicator of exposure and potential HIV infection among infants presenting at both well-child (immunization and community outreach) and sick-infant (nutrition and inpatient) entry points. To improve the identification of HIV-positive infants, nucleic acid-based testing should instead be considered in infants aged younger than 18 months.


Excreted Trypanosoma brucei proteins inhibit Plasmodium hepatic infection.
Malaria, a disease caused by Plasmodium parasites, remains a major threat to public health globally. It is the most common disease in patients with sleeping sickness, another parasitic illness, caused by Trypanosoma brucei. We have previously shown that a T. brucei infection impairs a secondary P. berghei liver infection and decreases malaria severity in mice. However, whether this effect requires an active trypanosome infection remained unknown. Here, we show that Plasmodium liver infection can also be inhibited by the serum of a mouse previously infected by T. brucei and by total protein lysates of this kinetoplastid. Biochemical characterisation showed that the anti-Plasmodium activity of the total T. brucei lysates depends on its protein fraction, but is independent of the abundant variant surface glycoprotein. Finally, we found that the protein(s) responsible for the inhibition of Plasmodium infection is/are present within a fraction of ~350 proteins that are excreted to the bloodstream of the host. We conclude that the defence mechanism developed by trypanosomes against Plasmodium relies on protein excretion. This study opens the door to the identification of novel antiplasmodial intervention strategies.


Art therapy based on appreciation of famous paintings and its effect on distress among cancer patients.
We aimed to evaluate the effectiveness of art therapy based on appreciation of famous paintings on the distress of cancer patients receiving radiotherapy. In particular, we focused on anxiety, depression, and cancer-related symptoms. Between October 2015 and February 2016, cancer patients receiving radiotherapy were recruited prospectively to participate in the art therapy based on famous painting appreciation. The art therapy took place in two parts comprising 4 sessions of famous painting appreciation and 4 sessions of creative artwork generation; these sessions were performed twice weekly over four weeks. Cancer-related distress was measured using the Hospital Anxiety and Depression Scale (HADS), Hamilton Depression Rating Scale (HDRS), and Edmonton Symptom Assessment Scale (ESAS) at three points: before the art therapy began, after the fourth session of art therapy, and after the eighth session. Of the 24 enrolled patients, 20 (83%) completed all eight sessions. We observed significant improvements in HADS anxiety and total scores over time according to linear mixed models with Bonferroni corrections (all p < 0.05). Furthermore, HDRS scores demonstrated significant decreases according to linear mixed models (p = 0.001). Fewer patients met the HADS or HDRS criteria for severe anxiety or depression after the intervention. We observed no changes in ESAS mean scores. Art therapy based on famous painting appreciation significantly improved cancer-related anxiety and depression and reduced the prevalence of severe anxiety and depression during cancer treatment.


Thoracic size-selective sampling of fibres: performance of four types of thoracic sampler in laboratory tests.
The counting of fibres on membrane filters could be facilitated by using size-selective samplers to exclude coarse particulate and fibres that impede fibre counting. Furthermore, the use of thoracic size selection would also remove the present requirement to discriminate fibres by diameter during counting. However, before thoracic samplers become acceptable for sampling fibres, their performance with fibres needs to be determined. This study examines the performance of four thoracic samplers: the GK2.69 cyclone, a Modified SIMPEDS cyclone, the CATHIA sampler (inertial separation) and the IOM thoracic sampler (porous foam pre-selector). The uniformity of sample deposit on the filter samples, which is important when counts are taken on random fields, was examined with two sizes of spherical particles (1 and 10 microm) and a glass fibre aerosol with fibres spanning the aerodynamic size range of the thoracic convention. Counts by optical microscopy examined fields on a set scanning pattern. Hotspots of deposition were detected for one of the thoracic samplers (Modified SIMPEDS with the 10 microm particles and the fibres). These hotspots were attributed to the inertial flow pattern near the port from the cyclone pre-separator. For the other three thoracic samplers, the distribution was similar to that on a cowled sampler, the current standard sampler for fibres. Aerodynamic selection was examined by comparing fibre concentration on thoracic samples with those measured on semi-isokinetic samples, using fibre size (and hence calculated aerodynamic diameter) and number data obtained by scanning electron microscope evaluation in four laboratories. The size-selection characteristics of three thoracic samplers (GK2.69, Modified SIMPEDS and CATHIA) appeared very similar to the thoracic convention; there was a slight oversampling (relative to the convention) for d(ae) < 7 microm, but that would not be disadvantageous for comparability with the cowled sampler. Only the IOM thoracic sampler tended to undersample the fibres relative to the thoracic convention. With the data divided into four classes based on fibre length, the size-selection characteristics appeared to be unaffected by fibre length for GK2.69, Modified SIMPEDS and CATHIA. Only the IOM thoracic sampler (with the foam selector) showed slightly lower selection for longer length classes of fibres. These results indicate that the tested samplers follow the thoracic sampling convention for fibres, and may be used to improve the quality and reliability of samples that are taken when there is likely to be significant background dust.


The Sesquiterpene Lactone Cynaropicrin Manifests Strong Cytotoxicity in Glioblastoma Cells U-87 MG by Induction of Oxidative Stress.
Cynaropicrin has shown a wide range of pharmacological properties, such as antitumor action. Here, we showed the inhibitory effect of Cyn on human glioblastoma cell U-87 MG growth. According to the IC50 values, Cyn 4, 8 and 10 µM displayed a significant cytotoxicity, as confirmed by the cell count and MTT assay. Furthermore, Cyn completely abolished the ability of U-87 MG to form colonies and induced drastic morphological changes. Interestingly, pretreatment with ROS scavenger N-acetylcysteine 3 mM reversed the cytotoxicity induced by Cyn 25 µM and preserved the cells by morphological changes. Therefore, oxidative stress induction was evaluated at low 8- and high 25-µM concentrations in U-87 MG, as demonstrated by the quantitative and qualitative analysis of ROS. A prolonged increase in ROS generation under Cyn 25 µM exposure was followed by the loss of the mitochondrial membrane potential in treated U-87 MG cells. An acute treatment with Cyn 25 µM induced Cyt c release, as revealed by immunofluorescence staining and the activation of cell death pathways, apoptosis and autophagy. On the other hand, chronic treatment with Cyn 8 µM induced senescence, as revealed by the increase in SA-β-Gal activity. Moreover, at this concentration, Cyn led to ERK dephosphorylation accompanied by a relevant reduction of the NF-κB p65 subunit. Finally, the combined effect of TMZ and Cyn resulted in synergistic cytotoxicity, as evaluated by the Bliss additivity model. The strong cytotoxicity of Cyn was also confirmed on IDH1 mutant U-87 MG cells and patient-derived IDH wild-type glioblastoma cell lines NULU and ZAR. In conclusion, given the high toxicity at minimal concentrations, the high inhibition of tumor cell growth and synergy with the standard drug for glioblastoma TMZ, Cyn could be proposed as a potential adjuvant for the treatment of glioblastoma.


Effect of drug ritual changes on schizophrenic patients.
The author evaluated the charts of 115 chronic schizophrenic patients who regularly attended a follow-up group to determine whether changes in long-standing medication regimens produced an increased dropout rate, decreased attendance, or psychotic decompensation. He found that although there were nonsignificant trends showing a relationship between medication change and group attendance for individual patients, there were no significant difference between group attendance and change or no change in medication routine for the group of patients as a whole. He concludes that necessary changes in medication routines for chronic schizophrenic patients are safe.


Individuality in cytochrome P450 expression and its association with the nephrotoxic and carcinogenic effects of chemicals.
The susceptibility of a tissue to the toxic and/or carcinogenic effects of chemicals is determined by a variety of factors, which include their rate of metabolic activation by the cytochrome P450-dependent monooxygenases. Individual differences in the levels of cytochrome P450 expression would be expected, and are known, to give rise to profound differences in toxicological response. Such effects are almost best exemplified by the sex differences observed in the toxic effects of a variety of nephrotoxins and carcinogens. In recent work, we have shown that in species such as the mouse and rat almost all cytochrome P450 enzymes in the kidney are sexually differentiated. This difference in cytochrome P450 regulation is mediated by testosterone and explains the large differences observed in the metabolic activation, toxicity and carcinogenicity of chloroform and possibly of other compounds such as ochratoxin A. In addition to hormonal or environmental influences on cytochrome P450 expression, genetic factors have also been shown to be important. In man, this is best exemplified by the genetic polymorphism observed in the metabolism of debrisoquine and approximately 25 other drugs. This genetic defect affects approximately 5-10% of the Caucasian population and has been associated with altered susceptibility to cancer. In this presentation, the development of a simple DNA-based assay to identify affected individuals is described. Use of this assay will allow clarification of the reported association of this genetic polymorphism to susceptibility to Balkan nephropathy and cancer.


Tris(pentafluorophenyl)borane as an efficient catalyst in the guanylation reaction of amines.
Tris(pentafluorophenyl)borane, [B(C6F5)3], has been used as an efficient catalyst in the guanylation reaction of amines with carbodiimide under mild conditions. A combined approach involving NMR spectroscopy and DFT calculations was employed to gain a better insight into the mechanistic features of this process. The results allowed us to propose a new Lewis acid-assisted Brønsted acidic pathway for the guanylation reaction. The process starts with the interaction of tris(pentafluorphenyl)borane and the amine to form the corresponding adduct, [(C6F5)3B-NRH2] , followed by a straightforward proton transfer to one of the nitrogen atoms of the carbodiimide, (i)PrN[double bond, length as m-dash]C[double bond, length as m-dash]N(i)Pr, to produce, in two consequent steps, a guanidine-borane adduct, [(C6F5)3B-NRC(N(i)PrH)2] . The rupture of this adduct liberates the guanidine product RNC(N(i)PrH)2 and interaction with additional amine restarts the catalytic cycle. DFT studies have been carried out in order to study the thermodynamic characteristics of the proposed pathway. Significant borane adducts with amines and guanidines have been isolated and characterized by multinuclear NMR in order to study the N-B interaction and to propose the existence of possible Frustrated Lewis Pairs. Additionally, the molecular structures of significant components of the catalytic cycle, namely 4-tert-butylaniline-[B(C6F5)3] adduct and both free and [B(C6F5)3]-bonded 1-(phenyl)-2,3-diisopropylguanidine, and respectively, have been established by X-ray diffraction.


In vivo GDF3 administration abrogates aging related muscle regeneration delay following acute sterile injury.
Tissue regeneration is a highly coordinated process with sequential events including immune cell infiltration, clearance of damaged tissues, and immune-supported regrowth of the tissue. Aging has a well-documented negative impact on this process globally; however, whether changes in immune cells per se are contributing to the decline in the body's ability to regenerate tissues with aging is not clearly understood. Here, we set out to characterize the dynamics of macrophage infiltration and their functional contribution to muscle regeneration by comparing young and aged animals upon acute sterile injury. Injured muscle of old mice showed markedly elevated number of macrophages, with a predominance for Ly6Chigh pro-inflammatory macrophages and a lower ratio of the Ly6Clow repair macrophages. Of interest, a recently identified repair macrophage-derived cytokine, growth differentiation factor 3 (GDF3), was markedly downregulated in injured muscle of old relative to young mice. Supplementation of recombinant GDF3 in aged mice ameliorated the inefficient regenerative response. Together, these results uncover a deficiency in the quantity and quality of infiltrating macrophages during aging and suggest that in vivo administration of GDF3 could be an effective therapeutic approach.


Human mesenchymal stromal cells attenuate graft-versus-host disease and maintain graft-versus-leukemia activity following experimental allogeneic bone marrow transplantation.
We sought to define the effects and underlying mechanisms of human, marrow-derived mesenchymal stromal cells (hMSCs) on graft-versus-host disease (GvHD) and graft-versus-leukemia (GvL) activity. Irradiated B6D2F1 mice given C57BL/6 BM and splenic T cells and treated with hMSCs had reduced systemic GvHD, donor T-cell expansion, and serum TNFα and IFNγ levels. Bioluminescence imaging demonstrated that hMSCs redistributed from lungs to abdominal organs within 72 hours, and target tissues harvested from hMSC-treated allogeneic BMT (alloBMT) mice had less GvHD than untreated controls. Cryoimaging more precisely revealed that hMSCs preferentially distributed to splenic marginal zones and regulated T-cell expansion in the white pulp. Importantly, hMSCs had no effect on in vitro cytotoxic T-cell activity and preserved potent GvL effects in vivo. Mixed leukocyte cultures containing hMSCs exhibited decreased T-cell proliferation, reduced TNFα, IFNγ, and IL-10 but increased PGE2 levels. Indomethacin and E-prostanoid 2 (EP2) receptor antagonisms both reversed while EP2 agonism restored hMSC-mediated in vitro T-cell suppression, confirming the role for PGE2 . Furthermore, cyclo-oxygenase inhibition following alloBMT abrogated the protective effects of hMSCs. Together, our data show that hMSCs preserve GvL activity and attenuate GvHD and reveal that hMSC biodistribute to secondary lymphoid organs wherein they attenuate alloreactive T-cell proliferation likely through PGE2 induction.


Evaluation of renal functions in children with congenital heart disease before and after cardiac angiography.
Radiocontrast nephrotoxicity, which has increased in incidence with widespread use of radiological methods in medicine, is a serious complication of radiocontrast materials. In this study, we have prospectively investigated whether children with cyanotic congenital heart disease are at risk for radiocontrast nephrotoxicity with the use of a nonionic low osmolar contrast agent. Thirty-five children (17 cyanotic and 18 acyanotic patients) who underwent diagnostic cardiac catheterization were subjects of the study. The age range was from five days to 13 years. The volume of contrast material was 3.11 +/- 1.37 ml/kg in cyanotic patients and 2.67 +/- 0.86 ml/kg in acyanotic patients. Blood samples and timed urine samples were taken from all patients 24 hours before and 48 hours after cardiac catheterization. Blood urea nitrogen, creatinine, sodium, and phosphorus in serum, and creatinine and N-acetyl-beta-D-glucosamine in urine were analyzed. There was not a statistically significant difference between the values before and after angiography. As a result, we could find no evidence of radiocontrast nephrotoxicity with the use of a nonionic contrast agent in cyanotic and acyanotic patients who underwent cardiac angiography.


[Incidence of lichens in the area of the Hamburg airport].
It was examined whether the co-additional effect of kerosene and benzine influences the air quality and the population of lichens. These plants were chosen since lichens serve as important indicators of air pollution. It was found that exhaust gases from a much frequented highway influence the number and the variation of species of lichens, whereas near the runway some less common species were found. These results show that air traffic exercises less influence on the lichen population than the road traffic emissions.


Strengthening Surveillance Systems for Malaria Elimination by Integrating Molecular and Genomic Data.
Unprecedented efforts in malaria control over the last 15 years have led to a substantial decrease in both morbidity and mortality in most endemic settings. However, these progresses have stalled over recent years, and resurgence may cause dramatic impact on both morbidity and mortality. Nevertheless, elimination efforts are currently going on with the objective of reducing malaria morbidity and mortality by 90% and malaria elimination in at least 35 countries by 2030. Strengthening surveillance systems is of paramount importance to reach those targets, and the integration of molecular and genomic techniques into routine surveillance could substantially improve the quality and robustness of data. Techniques such as polymerase chain reaction (PCR) and quantitative PCR (qPCR) are increasingly available in malaria endemic countries, whereas others such as sequencing are already available in a few laboratories. However, sequencing, especially next-generation sequencing (NGS), requires sophisticated infrastructure with adequate computing power and highly trained personnel for data analysis that require substantial investment. Different techniques will be required for different applications, and cost-effective planning must ensure the appropriate use of available resources. The development of national and sub-regional reference laboratories could help in minimizing the resources required in terms of equipment and trained staff. Concerted efforts from different stakeholders at national, sub-regional, and global level are needed to develop the required framework to establish and maintain these reference laboratories.


Stress-induced fractal rearrangement of the endothelial cell cytoskeleton causes apoptosis.
Apoptosis, a mechanism of cell death prominent in critical illnesses including disseminated inflammation and multiorgan dysfunction syndrome, is characterized by morphologic changes including cell shrinkage, condensation of organelles, blebbing, and chromatin fragmentation. These phenomena suggest substantial changes in cytoskeletal structure. We hypothesized that stress-induced apoptosis in endothelial cells is, in part, a consequence of a critical cytoskeletal rearrangement. Porcine aortic endothelial cells in culture, surrogates for the microvasculature in vivo, were exposed sequentially to Escherichia coli endotoxin (25 micrograms/mL; 18 hours) to induce the inflammatory response and then to sodium arsenite (160 mumol/L; 120 minutes) to induce the heat-shock response, a well-characterized model of stress-induced apoptosis. Laser confocal micrographs of fluorescein isothio-cyanate-labeled phalloidin-stained cells were analyzed to calculate the border fractal dimension of the cytoskeleton. Other cells were exposed to cytochalasin D, a fungal metabolite, which interferes with polymerization of actin from its globular to its filamentous form, and similarly were analyzed with respect to fractal dimension, viability (neutral red assay), and manner of death (annexin V fluorescence-activated cell scanning analysis). Induction of the inflammatory or heat-shock responses caused subtle and distinct rearrangement of the actin cytoskeleton. When these stimuli were applied in sequence, a synergistic interaction led to profound cytoskeletal collapse. Reversal of the sequence did not induce the cytoskeletal disruption. Cytochalasin D alone induced a dose-dependent cytoskeletal collapse indistinguishable from that caused by the acute phase-heat shock sequence that caused cell death by apoptosis. The effect of lower doses of Cytochalasin D could be potentiated by subsequent induction of the heat-shock response. Sequential stresses that mimic pathophysiologic "two-hit" stimuli induce a characteristic fractal rearrangement of the actin cytoskeleton. Because cytochalasin D-induced rearrangement of this cytoskeleton produced apoptosis indistinguishable from the stress-induced apoptosis, we conclude that the cytoskeletal rearrangement is likely a critical event in the pathway to apoptosis. This disruption of intracellular interconnections mirrors endotoxin-induced disruption in signals among organs and supports the mechanistic hypothesis that multiorgan dysfunction syndrome generally reflects disruption of signals and connections at several levels of biologic organization.


Pre-analytical stability of serum biomarkers for neurological disease: neurofilament-light, glial fibrillary acidic protein and contactin-1.
Neurofilament-light (NfL), glial fibrillary acidic protein (GFAP) and contactin-1 (CNTN1) are blood-based biomarkers that could contribute to monitoring and prediction of disease and treatment outcomes in neurological diseases. Pre-analytical sample handling might affect results, which could be disease-dependent. We tested common handling variations in serum of volunteers as well as in a defined group of patients with multiple sclerosis (pwMS). Sample sets from 5 pwMS and 5 volunteers at the outpatient clinic were collected per experiment. We investigated the effect of the following variables: collection tube type, delayed centrifugation, centrifugation temperature, delayed storage after centrifugation and freeze-thawing. NfL and GFAP were measured by Simoa and CNTN1 by Luminex. A median recovery of 90-110% was considered stable. For most pre-analytical variables, serum NfL and CNTN1 levels remained unaffected. In the total group, NfL levels increased (121%) after 6 h of delay at 2-8 °C until centrifugation, while no significant changes were observed after 24 h delay at room temperature (RT). In pwMS specifically, CNTN1 levels increased from additional freeze-thaw cycles number 2 to 4 (111%-141%), whereas volunteer levels remained stable. GFAP showed good stability for all pre-analytical variables. Overall, the serum biomarkers tested were relatively unaffected by variations in sample handling. For serum NfL, we recommend storage at RT before centrifugation at 2-8 °C up to 6 h or at RT up to 24 h. For serum CNTN1, we advise a maximum of two freeze-thaw cycles. Our results confirm and expand on recently launched consensus standardized operating procedures.


Promising strategies for obesity prevention and treatment within American Indian communities.
In the United States, obesity prevention and treatment programs are dominated by individual-focused strategies. Yet epidemic proportions of obesity and high failure and dropout rates of conventional efforts prompt consideration of alternate approaches. A nonsystematic review of American Indian (AI) health promotion programs highlights a group-focused approach using three primary and two supportive strategies. These strategies are outlined as: (a) build and reinforce social cohesion and collective efficacy, (b) use the motivating force of friendly competition, and (c) aspire to change local norms and policies through assuring high visibility of alternate behaviors and engaging formal and informal leaders. Although evidence to demonstrate the impact of these efforts is limited and not currently available to public health audiences, AI communities' sustained acceptance and continued involvement in the programs suggest that these strategies are worthy of review and consideration by public health practitioners addressing obesity on a national level.


English-language videos on YouTube as a source of information on self-administer subcutaneous anti-tumour necrosis factor agent injections.
The aim of this study was to evaluate the reliability, content, and quality of videos for patients available on YouTube for learning how to self-administer subcutaneous anti-tumour necrosis factor (TNF) injections. We searched for the terms Humira injection, Enbrel injection, Simponi injection, and Cimzia injection. Videos were categorised as useful information, misleading information, useful patient opinion, and misleading patient opinion by two physicians. Videos were rated for quality on a 5-point global quality scale (GQS; 1 = poor quality, 5 = excellent quality) and reliability and content using the 5-point DISCERN scale (higher scores represent greater reliability and more comprehensive videos). Of the 142 English videos, 24 (16.9%) videos were classified as useful information, 6 (4.2%) as misleading information, 47 (33.1%) as useful patient opinion, and 65 (45.8%) as misleading patient opinion. Useful videos were the most comprehensive and had the highest reliability and quality scores. The useful information and useful patient opinion videos had the highest numbers of views per day (median 8.32, IQR: 3.40-14.28 and 5.46, IQR: 3.06-14.44), as compared with 2.32, IQR: 1.63-6.26 for misleading information videos and 2.15, IQR: 1.17-7.43 for misleading patient opinion videos (p = 0.001). Almost all (91.5%) misleading videos were uploaded by individual users. There are a substantial number of English-language YouTube videos, with high quality, and rich content and reliability that can be sources of information on proper technique of anti-TNF self-injections. Physicians should direct patients to the reliable resources of information and educate them in online resource assessment, thereby improving treatment outcomes.


Enteric Pathogens and Reactive Arthritis: Systematic Review and Meta-Analyses of Pathogen-Associated Reactive Arthritis.
The objective of this systematic review and meta-analysis was to estimate the proportion of postinfectious reactive arthritis (ReA) after bacterial enteric infection from one of four selected pathogens. We collected studies from PubMed, Web of Science, and Embase, which assessed the proportion of postinfectious ReA published from January 1, 2000 to April 1, 2018. Papers were screened independently by title, abstract, and full text; papers in English, Spanish, and Portuguese utilizing a case-control (CC) or cohort study design, with a laboratory confirmed or probable acute bacterial enteric infection and subsequent ReA, were included. The proportion of ReA cases was pooled between and across pathogens. Factors that can induce study heterogeneity were explored using univariate meta-regression, including region, sample size, study design, and ReA case ascertainment. Twenty-four articles were included in the final review. The estimated percentage of cases across studies describing Campylobacter-associated ReA (n = 11) was 1.71 (95% confidence interval [CI] 0.49-5.84%); Salmonella (n = 17) was 3.9 (95% CI 1.6-9.1%); Shigella (n = 6) was 1.0 (95% CI 0.2-4.9%); and Yersinia (n = 7) was 3.4 (95% CI 0.8-13.7%). Combining all four pathogens, the estimated percentage of cases that developed ReA was 2.6 (95% CI 1.5-4.7%). Due to high heterogeneity reflected by high I2 values, results should be interpreted with caution. However, the pooled proportion developing ReA from studies with sample sizes (N) <1000 were higher compared with N > 1000 (6% vs. 0.3%), retrospective cohort studies were lower (1.1%) compared with CC or prospective cohorts (6.8% and 5.9%, respectively), and those where ReA cases are identified through medical record review were lower (0.3%) than those identified by a specialist (3.9%) or self-report (12%). The estimated percentage of people who developed ReA after infection with Campylobacter, Salmonella, Shigella, or Yersinia is relatively low (2.6). In the United States, this estimate would result in 84,480 new cases of ReA annually.


Optimising superoscillatory spots for far-field super-resolution imaging.
Optical superoscillatory imaging, allowing unlabelled far-field super-resolution, has in recent years become reality. Instruments have been built and their super-resolution imaging capabilities demonstrated. The question is no longer whether this can be done, but how well: what resolution is practically achievable? Numerous works have optimised various particular features of superoscillatory spots, but in order to probe the limits of superoscillatory imaging we need to simultaneously optimise all the important spot features: those that define the resolution of the system. We simultaneously optimise spot size and its intensity relative to the sidebands for various fields of view, giving a set of best compromises for use in different imaging scenarios. Our technique uses the circular prolate spheroidal wave functions as a basis set on the field of view, and the optimal combination of these, representing the optimal spot, is found using a multi-objective genetic algorithm. We then introduce a less computationally demanding approach suitable for real-time use in the laboratory which, crucially, allows independent control of spot size and field of view. Imaging simulations demonstrate the resolution achievable with these spots. We show a three-order-of-magnitude improvement in the efficiency of focusing to achieve the same resolution as previously reported results, or a 26 % increase in resolution for the same efficiency of focusing.


Slippery when wet: phylogeny and character evolution in the gelatinous cyanobacterial lichens (Peltigerales, Ascomycetes).
Many lichen fungi form symbioses with filamentous Nostoc cyanobacteria, which cause the lichen to swell and become extremely gelatinous when moist. Within the Lecanoromycetes, such gelatinous lichens are today mainly classified in the Collemataceae (Peltigerales, Ascomycota). We performed Bayesian MCMC, maximum likelihood, and maximum parsimony analyses of three independent markers (mtSSU rDNA, nuLSU rDNA, and RPB1), to improve our understanding of the phylogeny and classification in the Peltigerales, as well as the evolution of morphological characters that have been used for classification purposes in this group. The Collemataceae and the non-gelatinous Pannariaceae are paraphyletic but can be re-circumscribed as monophyletic if Leciophysma, Physma, Ramalodium and Staurolemma are transferred to the Pannariaceae. The gelatinous taxa transferred to the Pannariaceae deviate from other Collemataceae in having simple ascospores, and several also have a ring-shaped exciple as in other Pannariaceae, rather than the disc-shaped exciple found in the typical Collemataceae. Both Collema and Leptogium are non-monophyletic. The re-circumscribed Collemataceae shares a distinct ascus type with the sister group Placynthiaceae and the Coccocarpiaceae, whereas Pannariaceae includes a variety of structures. All Pannariaceae have one-celled ascospores, whereas all Collemataceae have two- or multi-celled spores. Reconstructions of the number of character state transformations in exciple structure, thallus gelatinosity, and ascus apex structure indicate that the number of transformations is distinctly higher than the minimum possible. Most state transformations in the exciple took place from a ring-shaped to a disc-shaped exciple. Depending on the reconstruction method, most or all transformations in thallus structure took place from a non-gelatinous to a gelatinous thallus. Gains and losses of internal structures in the ascus apex account for all or a vast majority of the number of transformations in the ascus, whereas direct transformations between asci with internal structures appear to have been rare.


Activation of skeletal muscle-resident glial cells upon nerve injury.
Here, we report on the identification of Itga7-expressing muscle-resident glial cells activated by loss of neuromuscular junction (NMJ) integrity. Gene expression analysis at the bulk and single-cell level revealed that these cells are distinct from Itga7-expressing muscle satellite cells. We show that a selective activation and expansion of Itga7+ glial cells occur in response to muscle nerve lesion. Upon activation, muscle glial-derived progenies expressed neurotrophic genes, including nerve growth factor receptor, which enables their isolation by FACS. We show that activated muscle glial cells also expressed genes potentially implicated in extracellular matrix remodeling at NMJs. We found that tenascin C, which was highly expressed by muscle glial cells, activated upon nerve injury and preferentially localized to NMJ. Interestingly, we observed that the activation of muscle glial cells by acute nerve injury was reversible upon NMJ repair. By contrast, in a mouse model of ALS, in which NMJ degeneration is progressive, muscle glial cells steadily increased over the course of the disease. However, they exhibited an impaired neurotrophic activity, suggesting that pathogenic activation of glial cells may be implicated in ALS progression.


Treatment of polymyositis and dermatomyositis.
Polymyositis and dermatomyositis are serious inflammatory muscle disorders which may present life-threatening complications. It is important to recognise and treat the condition in the early stages of the disease. Corticosteroids remain the mainstay of treatment, and their resistance and other forms of immunotherapy are discussed.


Characterization of Human Duodenal Fluids in Fasted and Fed State Conditions.
This work provides an elaborate characterization of human intestinal fluids (HIF) collected in fasted- and fed-state conditions. HIF from 20 healthy volunteers (10 M/F) were aspirated by intubation near the ligament of Treitz in a time-dependent manner (10-min intervals) and characterized for pH, bile salts, phospholipids, cholesterol, triacylglycerides (TAG), diacylglycerides (DAG), monoacylglycerides (MAG), free fatty acids (FFA), pancreatic lipase, phospholipase A2, and nonspecific esterase activity. For almost all parameters, a food-induced effect was observed. Results were characterized by a high variability, as illustrated by the broad ranges observed for each parameter: pH (fasted: 3.4-8.3; fed: 4.7-7.1), bile salts (fasted: 0.03-36.18 mM; fed: 0.74-86.14 mM), phospholipids (fasted: 0.01-6.33 mM; fed: 0.16-14.39 mM), cholesterol (fasted: 0-0.48 mM; fed: 0-3.29 mM), TAG (fed: 0-6.76 mg/mL), DAG (fed: 0-3.64 mg/mL), MAG (fasted: 0-1.09 mg/mL; fed: 0-11.36 mg/mL), FFA (fasted: 0-3.86 mg/mL; fed: 0.53-15.0 mg/mL), pancreatic lipase (fasted: 26-86 g/mL; fed: 146-415 g/mL), phospholipase A2 (fasted: 3-6 ng/mL; fed: 4.3-27.7 ng/mL), and nonspecific esterase activity (fasted: 270-4900 U/mL; fed: 430-4655 U/mL). This comprehensive overview may serve as reference data for physiologically based pharmacokinetic modeling and the optimization of biorelevant simulated intestinal fluids for the use in in vitro dissolution, solubility, and permeability profiling.


[Medical target-setting].
Typical situations in the work of a primary care doctor are considered, viz. the purpose of visits familiarizing the doctor with the patient and his (her) family, drafting the patient's medical chart and the family's medical file folder, frequency and purpose of the patient's visits to the doctor's office in acute clinical condition, logical structure of primary care for planning active case detection.


[Modernization of organization base of technical service and repair of medical technique].
The article presents organizational aspects of technical service and repair of medical technique, used by medical service of the Armed Forces. Was analyzed order of bringing into service of new medical technique, reasons of untimely solving of this task. Was presented a systemic structure of forces and measures on technical service and repair of medical technique, which includes 3 levels: I--Center, II--military region, III--territorial area of responsibility. In details were observed tasks of every level of responsibility.


The Genomic and Phenotypic Landscape of Ichthyosis: An Analysis of 1000 Kindreds.
Ichthyoses are clinically and genetically heterogeneous disorders characterized by scaly skin. Despite decades of investigation identifying pathogenic variants in more than 50 genes, clear genotype-phenotype associations have been difficult to establish. To expand the genotypic and phenotypic spectra of ichthyosis and delineate genotype-phenotype associations. This cohort study recruited an international group of individuals with ichthyosis and describes characteristic and distinguishing features of common genotypes, including genotype-phenotype associations, during a 10-year period from June 2011 to July 2021. Participants of all ages, races, and ethnicities were included and were enrolled worldwide from referral centers and patient advocacy groups. A questionnaire to assess clinical manifestations was completed by those with a genetic diagnosis. Genetic analysis of saliva or blood DNA, a phenotyping questionnaire, and standardized clinical photographs. Descriptive statistics, such as frequency counts, were used to describe the cases in the cohort. Fisher exact tests identified significant genotype-phenotype associations. Results were reported for 1000 unrelated individuals enrolled from around the world (mean [SD] age, 50.0 [34.0] years; 524 [52.4%] were female, 427 [42.7%] were male, and 49 [4.9%] were not classified); 75% were from the US, 12% from Latin America, 4% from Canada, 3% from Europe, 3% from Asia, 2% from Africa, 1% from the Middle East, and 1% from Australia and New Zealand. A total of 266 novel disease-associated variants in 32 genes were identified among 869 kindreds. Of these, 241 (91%) pathogenic variants were found through multiplex amplicon sequencing and 25 (9%) through exome sequencing. Among the 869 participants with a genetic diagnosis, 304 participants (35%) completed the phenotyping questionnaire. Analysis of clinical manifestations in these 304 individuals revealed that pruritus, hypohydrosis, skin pain, eye problems, skin odor, and skin infections were the most prevalent self-reported features. Genotype-phenotype association analysis revealed that the presence of a collodion membrane at birth (odds ratio [OR], 6.7; 95% CI, 3.0-16.7; P < .001), skin odor (OR, 2.8; 95% CI, 1.1-6.8; P = .02), hearing problems (OR, 2.9; 95% CI, 1.6-5.5; P < .001), eye problems (OR, 3.0; 95% CI, 1.5-6.0; P < .001), and alopecia (OR, 4.6; 95% CI, 2.4-9.0; P < .001) were significantly associated with TGM1 variants compared with other ichthyosis genotypes studied. Skin pain (OR, 6.8; 95% CI, 1.6-61.2; P = .002), odor (OR, 5.7; 95% CI, 2.0-19.7; P < .001), and infections (OR, 3.1; 95% CI, 1.4-7.7; P = .03) were significantly associated with KRT10 pathogenic variants compared with disease-associated variants in other genes that cause ichthyosis. Pathogenic variants were identified in 869 (86.9%) participants. Most of the remaining individuals had unique phenotypes, enabling further genetic discovery. This cohort study expands the genotypic and phenotypic spectrum of ichthyosis, establishing associations between clinical manifestations and genotypes. Collectively, the findings may help improve clinical assessment, assist with developing customized management plans, and improve clinical course prognostication.


Iterative image reconstruction using prior knowledge.
A method is proposed to reconstruct signals from incomplete data. The method, which can be interpreted both as a discrete implementation of the so-called prior discrete Fourier transform (PDFT) spectral estimation technique and as a variant of the algebraic reconstruction technique, allows one to incorporate prior information about the reconstructed signal to improve the resolution of the signal estimated. The context of diffraction tomography and image reconstruction from samples of the far-field scattering amplitude are used to explore the performance of the method. On the basis of numerical computations, the optimum choice of parameters is determined empirically by comparing image reconstructions of the noniterative PDFT algorithm and the proposed iterative scheme.


The polyphenols resveratrol and S17834 prevent the structural and functional sequelae of diet-induced metabolic heart disease in mice.
Diet-induced obesity is associated with metabolic heart disease characterized by left ventricular hypertrophy and diastolic dysfunction. Polyphenols such as resveratrol and the synthetic flavonoid derivative S17834 exert beneficial systemic and cardiovascular effects in a variety of settings including diabetes mellitus and chronic hemodynamic overload. We characterized the structural and functional features of a mouse model of diet-induced metabolic syndrome and used the model to test the hypothesis that the polyphenols prevent myocardial hypertrophy and diastolic dysfunction. Male C57BL/6J mice were fed a normal diet or a diet high in fat and sugar (HFHS) with or without concomitant treatment with S17834 or resveratrol for up to 8 months. HFHS diet-fed mice developed progressive left ventricular hypertrophy and diastolic dysfunction with preservation of systolic function in association with myocyte hypertrophy and interstitial fibrosis. In HFHS diet-fed mice, there was increased myocardial oxidative stress with evidence of oxidant-mediated protein modification via tyrosine nitration and 4-OH-2-nonenol adduction. HFHS diet-fed mice also exhibited increases in plasma fasting glucose, insulin, and homeostasis model assessment of insulin resistance indicative of insulin resistance. Treatment with S17834 or resveratrol prevented left ventricular hypertrophy and diastolic dysfunction. For S17834, these beneficial effects were associated with decreases in oxidant-mediated protein modifications and hyperinsulinemia and increased plasma adiponectin. Resveratrol and S17834 administered concurrently with a HFHS diet prevent the development of left ventricular hypertrophy, interstitial fibrosis, and diastolic dysfunction. Multiple mechanisms may contribute to the beneficial effects of the polyphenols, including a reduction in myocardial oxidative stress and related protein modifications, amelioration of insulin resistance, and increased plasma adiponectin. The polyphenols resveratrol and S17834 may be of value in the prevention of diet-induced metabolic heart disease.


Sustaining the Canadian Nursing Workforce: Targeted Evidence-Based Reactive Solutions in Response to the Ongoing Crisis.
Inadequate staffing, excessive workloads, endemic violence and unhealthy workplaces are some of the challenges facing Canadian nurses. Leaving these issues unaddressed has had pernicious impacts on the nursing workforce: thousands of nurses across Canada have been suffering from extreme stress, anxiety and burnout, leading many of them to leave their current jobs and, for some, the profession of nursing altogether. We conducted a comprehensive yet rapid review of evidence-based solutions from the peer-reviewed and policy literature, stakeholder dialogues and member surveys commissioned by the Canadian Federation of Nurses Unions that could be implemented and scaled across Canada. Our findings support coordinated series of collectively planned, carefully sequenced and evidence-based interventions to retain, return, integrate and recruit nurses targeted to support the nursing workforce from training to early-, mid- and late-career stages. The implementation of these reactive solution bundles will also enhance the quality of healthcare services and, more broadly, the healthcare system.


Auritidibacter ignavus, an Emerging Pathogen Associated with Chronic Ear Infections.
We describe detection of the previously rarely reported gram-positive bacterium Auritidibacter ignavus in 3 cases of chronic ear infections in Germany. In all 3 cases, the patients had refractory otorrhea. Although their additional symptoms varied, all patients had an ear canal stenosis and A. ignavus detected in microbiologic swab specimens. A correct identification of A. ignavus in the clinical microbiology laboratory is hampered by the inability to identify it by using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Also, the bacterium might easily be overlooked because of its morphologic similarity to bacterial species of the resident skin flora. We conclude that a high index of suspicion is warranted to identify A. ignavus and that it should be particularly considered in patients with chronic external otitis who do not respond clinically to quinolone ear drop therapy.


Vibrio fischeri and Escherichia coli adhesion tendencies towards photolithographically modified nanosmooth poly (tert-butyl methacrylate) polymer surfaces.
This study reports the adhesion behavior of two bacterial species, Vibrio fischeri and Escherichia coli, to the photoresistant poly(tert-butyl methacrylate) (P(tBMA)) polymer surface. The data has demonstrated that ultraviolet irradiation of P(tBMA) was able to provide control over bacterial adhesion tendencies. Following photolithography, several of the surface characteristics of P(tBMA) were found to be altered. Atomic force microscopy analysis indicated that photolithographically modified P(tBMA) (henceforth termed 'modified polymer') appeared as a 'nanosmooth' surface with an average surface roughness of 1.6 nm. Although confocal laser scanning microscopy and scanning electron microscopy analysis clearly demonstrated that V. fischeri and E. coli presented largely different patterns of attachment in order to adhere to the same surfaces, both species exhibited a greater adhesion propensity towards the 'nanosmooth' surface. The adhesion of both species to the modified polymer surface appeared to be facilitated by an elevated production of extracellular polymeric substances when in contact with the substrate.