DNA microarray technology and application.
DNA microarrays (collections of DNA probes arranged on a shared base) have recently enlarged the spectrum of commercially available laboratory-ready kits in molecular biology. They are powerful new tools for the investigation of global changes in gene expression profiles in cells and tissues. Their assembly process is automatized and the DNA microarrays are further miniaturized. The DNA microarrays are used in search for various specific genes (e.g. connected with an infectious agent) or in gene polymorphism and expression analysis. They will be widely used to investigate expression of various genes connected with various diseases in order to find causes of these diseases and to enable their accurate treatment. Since the DNA microarray assembly technology has been based on methods widely used in the semiconductor industry, we can expect a rapid onset of the routine use of this revolutionary device.


Ontogeny of mu- and kappa-opiate receptor control of the hypothalamo-pituitary-adrenal axis in rats.
The purpose of the present study was to determine the relative ontogeny of mu- and kappa-opiate receptor control of the hypothalamo-pituitary-adrenal (HPA) axis in rats. The ability of the mu-agonist morphine and the kappa-agonist U-50,488 to stimulate the HPA axis was determined by evaluating ACTH and corticosterone (CS) secretion in developing rat pups. Morphine elicited marked rises in both ACTH and CS secretion in 10-day-old rats, and these increases were maximal from 30-60 min after drug administration. Both morphine and U50,488H caused a dose-related rise in CS secretion that was blocked by the synthetic glucocorticoid dexamethasone. The mu-opiate antagonist beta-funaltrexamine blocked the morphine-induced rise in CS secretion, and the kappa-antagonist norbinaltorphimine blocked the action of U50,488H. While a maximal dose of U50,488H (1 mg/kg) elicited a significant rise in CS secretion as early as postnatal day 2, significant effects of a maximal dose of morphine (5 mg/kg) were not observed until day 5. The effects of both drugs were significantly blunted during the stress-hyporesponsive period from days 5-15. The results of this study demonstrate that significant opiate receptor control of HPA function can be demonstrated early in postnatal development, even before the onset of the stress-hyporesponsive period. In addition, these data suggest that kappa-receptor control is functional before mu-receptor control of HPA function.


Shock wave lithotripsy, for the treatment of kidney stones, results in changes to routine blood tests and novel biomarkers: a prospective clinical pilot-study.
The number of patients undergoing shock wave lithotripsy (SWL) for kidney stones is increasing annually, and as such the development of post-operative complications, such as haematuria and acute kidney injury (AKI) following SWL, is likely to increase. The aim of the study was to evaluate changes in routine blood and novel biomarkers following SWL, for the treatment of kidney stones. Twelve patients undergoing SWL for solitary unilateral kidney stones were recruited. From patients (8 males and 4 females) aged between 31 and 72 years (median 43 years), venous blood samples were collected pre-operatively (baseline), at 30, 120 and 240 min post-operatively. Routine blood tests were performed using a Sysmex XE-5000, and Beckman Coulter AU5800 and AU680 analysers. NGAL, IL-18, IL-6, TNF-α, IL-10 and IL-8 concentrations were determined using commercially available ELISA kits. Significant (p ≤ 0.05) changes were observed in several blood parameters following SWL. NGAL concentration significantly increased, with values peaking at 30 min post-treatment (p = 0.033). Although IL-18 concentration increased, these changes were not significant (p = 0.116). IL-6 revealed a statistically significant rise from pre-operative up to 4 h post-operatively (p < 0.001), whilst TNF-α significantly increased, peaking at 30 min post-SWL (p = 0.05). There were no significant changes to IL-10 and IL-8 concentrations post-SWL (p > 0.05). Changes to routine blood tests and specific biomarkers, in the future, may be more useful for clinicians. In turn, identification of a panel of biomarkers could provide valuable data on "normal" physiological response after lithotripsy. Ultimately, studies could be expanded to identify or predict those patients at increased risk of developing post-operative complications, such as acute kidney injury or. These studies, however, need validating involving larger cohorts.


Analysis of genetic variation in Akt2/PKB-beta in severe insulin resistance, lipodystrophy, type 2 diabetes, and related metabolic phenotypes.
We previously reported a family in which a heterozygous missense mutation in Akt2 led to a dominantly inherited syndrome of insulin-resistant diabetes and partial lipodystrophy. To determine whether genetic variation in AKT2 plays a broader role in human metabolic disease, we sequenced the entire coding region and splice junctions of AKT2 in 94 unrelated patients with severe insulin resistance, 35 of whom had partial lipodystrophy. Two rare missense mutations (R208K and R467W) were identified in single individuals. However, insulin-stimulated kinase activities of these variants were indistinguishable from wild type. In two large case-control studies (total number of participants 2,200), 0 of 11 common single nucleotide polymorphism (SNPs) in AKT2 showed significant association with type 2 diabetes. In a quantitative trait study of 1,721 extensively phenotyped individuals from the U.K., no association was found with any relevant intermediate metabolic trait. In summary, although heterozygous loss-of- function mutations in AKT2 can cause a syndrome of severe insulin resistance and lipodystrophy in humans, such mutations are uncommon causes of these syndromes. Furthermore, genetic variation in and around the AKT2 locus is unlikely to contribute significantly to the risk of type 2 diabetes or related intermediate metabolic traits in U.K. populations.


The Toxoplasma protein phosphatase 6 catalytic subunit (TgPP6C) is essential for cell cycle progression and virulence.
Protein phosphatases are post-translational regulators of Toxoplasma gondii proliferation, tachyzoite-bradyzoite differentiation and pathogenesis. Here, we identify the putative protein phosphatase 6 (TgPP6) subunits of T. gondii and elucidate their role in the parasite lytic cycle. The putative catalytic subunit TgPP6C and regulatory subunit TgPP6R likely form a complex whereas the predicted structural subunit TgPP6S, with low homology to the human PP6 structural subunit, does not coassemble with TgPP6C and TgPP6R. Functional studies showed that TgPP6C and TgPP6R are essential for parasite growth and replication. The ablation of TgPP6C significantly reduced the synchronous division of the parasite's daughter cells during endodyogeny, resulting in disordered rosettes. Moreover, the six conserved motifs of TgPP6C were required for efficient endodyogeny. Phosphoproteomic analysis revealed that ablation of TgPP6C predominately altered the phosphorylation status of proteins involved in the regulation of the parasite cell cycle. Deletion of TgPP6C significantly attenuated the parasite virulence in mice. Immunization of mice with TgPP6C-deficient type I RH strain induced protective immunity against challenge with a lethal dose of RH or PYS tachyzoites and Pru cysts. Taken together, the results show that TgPP6C contributes to the cell division, replication and pathogenicity in T. gondii.


Implementation of a virtual laboratory for training on sound insulation testing and uncertainty calculations in acoustic tests.
This paper describes a methodology and case study for the implementation of educational virtual laboratories for practice training on acoustic tests according to international standards. The objectives of this activity are (a) to help the students understand and apply the procedures described in the standards and (b) to familiarize the students with the uncertainty in measurement and its estimation in acoustics. The virtual laboratory will not focus on the handling and set-up of real acoustic equipment but rather on procedures and uncertainty. The case study focuses on the application of the virtual laboratory for facade sound insulation tests according to ISO 140-5:1998 (International Organization for Standardization, Geneva, Switzerland, 1998), and the paper describes the causal and stochastic models and the constraints applied in the virtual environment under consideration. With a simple user interface, the laboratory will provide measurement data that the students will have to process to report the insulation results that must converge with the "virtual true values" in the laboratory. The main advantage of the virtual laboratory is derived from the customization of factors in which the student will be instructed or examined (for instance, background noise correction, the detection of sporadic corrupted observations, and the effect of instrument precision).


[Publication practice and policy for disciplinary proceedings regarding health care in the Netherlands].
To provide insight into practice and policy regarding the publication of disciplinary verdicts in Dutch health care. Descriptive. The study covered all of the verdicts issued by disciplinary boards and courts of justice that were published in the Dutch the Staats-courant (Government Gazette) during the period 1995-2002. The following attributes were recorded for each verdict: year of publication and verdict, the disciplinary board concerned, the accused professional, the type of complainant, the nature of the complaint and verdict, and the journals that were offered the decision. The published verdicts were related to the total number of verdicts and cases during the study period. Questionnaires were used to investigate the publication policy of the disciplinary boards. A total of 4% of all verdicts were published in the Dutch Government Gazette (323/8902). The central disciplinary board decided to publish more often than the regional disciplinary boards (8% and 2%, respectively). There were considerable differences between the various regional disciplinary boards (range: 1-5%). Per professional group the percentage of verdicts in cases that were published ranged from 2% to 23%. The decisions were offered to over 20 journals, but mainly to the Tijdschrift voor Gezondheidsrecht [Dutch Journal of Health Law] (TvGR; 92%) and Medisch Contact [Medical Contact] (MC; 88%). The TvGR published almost two-thirds of the verdicts offered (63%), and the MC published almost three-quarters (74%). In the decision to publish, the disciplinary boards differed in their interpretation of the concept of 'general importance'. If disciplinary proceedings are to achieve the desired quality-promoting effect on professional practice, then more attention will need to be paid to the publication policy, and the disciplinary boards will have to develop a joint code of practice. More verdicts could be published, also in professional journals.


Quantitative cone-beam computed tomography evaluation of premaxilla and symphysis in Class I and Class III malocclusions.
In order to compare the premaxilla and symphysis in Class I and Class III individuals, we measured the following parameters on CBCT scans: incisor inclination, symphyseal morphology, upper alveolar process morphology, symphyseal and maxillary leeway space, as well as anterior and posterior maxillary and mandibular bone thickness at various points below the cementoenamel junction. CBCT scans were taken of 62 individuals (41 females and 21 males) of ages ranging between 11 and 53years (mean 23.26). The dentoskeletal relationship and alveolar process morphology (thickness) were evaluated on axial slices of the upper and lower jaws, and Student's parametric t-test for paired data was used to compare Class I and Class III values within different facial types. Many significant differences in measurements between the two facial types were noted in terms of tooth inclination, alveolar thickness, and leeway space. Our findings confirm that Class III patients have little periodontal support at the upper and lower incisors, irrespective of the amount of symphyseal leeway space available, and any tooth movement must therefore be carefully planned and monitored.


Water, waste, and well-being: a multicountry study.
Data collected in the late 1980s from eight countries in Sub-Saharan Africa (Burundi, Ghana, Togo, and Uganda), Asia/North Africa (Sri Lanka and Morocco), and the Americas (Bolivia and Guatemala) were combined and analyzed to test whether incremental health effects regarding diarrhea and nutritional status result from incremental improvements in water and sanitation conditions. Rural (n = 11,992) and urban (n = 4,888) samples were analyzed separately. Optimal (i.e., on the premises) and intermediate (improved public water) water supplies were compared with unimproved water conditions. Optimal (flush toilets or water-seal-latrines) and intermediate (latrines) sanitation levels were compared with unimproved sanitation. Nationally representative (random) samples of ever-married women age 15-49 years, with or without children, were interviewed in all countries, and children aged 3-36 months with available weight and height data were included in the analyses. Multiple linear regression controlled for household, maternal, and child-level variables; in addition, dummy variables were included for each country. Improvements in sanitation resulted in less diarrhea and in taller and heavier children with each of the three levels of water supply. Incremental benefits in sanitation were associated with less diarrhea and with additional increases in the weights and heights of children. The effects of improved sanitation were greater among urban dwellers than among rural dwellers. Health benefits from improved water were less pronounced than those for sanitation. Benefits from improved water occurred only when sanitation was improved and only when optimal water was present. These findings suggest that public health intervention should balance epidemiologic data with the cost of services and the demand for water. There should be efforts to develop compatible technologies so that incremental improvements in service can be made.


Redd1 is a novel marker of testis development but is not required for normal male reproduction.
In an effort to identify novel candidate genes involved in testis determination, we previously used suppression subtraction hybridisation PCR on male and female whole embryonic (12.0-12.5 days post coitum) mouse gonads. One gene to emerge from our screen was Redd1. In the current study, we demonstrate by whole-mount in situ hybridisation that Redd1 is differentially expressed in the developing mouse gonad at the time of sex determination, with higher expression in testis than ovary. Furthermore, Redd1 expression was first detected as Sry expression peaks, immediately prior to morphological sex determination, suggesting a potential role for Redd1 during testis development. To determine the functional importance of this gene during testis development, we generated Redd1-deficient mice. Morphologically, Redd1-deficient mice were indistinguishable from control littermates and showed normal fertility. Our results show that Redd1 alone is not required for testis development or fertility in mice. The lack of a male reproductive phenotype in Redd1 mice may be due to functional compensation by the related gene Redd2.


Cross-cultural invariance of the factor structure of the Schizotypal Personality Questionnaire across Spanish and American college students.
The main goal of this study was to examine the cross-cultural invariance of the factor structure of the Schizotypal Personality Questionnaire (SPQ) (Raine, 1991) in two large samples of Spanish and American young adults. The final sample was made up of 2313 college students (508 men, 22%). Their mean age was 20.5 years (S.D.=3.2). The results indicated that the Stefanis et al. (2004) four-factor model yielded the best goodness-of-fit indices compared to alternative models. Moreover, the results support configural, metric, and partial measurement invariance of the covariances of the SPQ across the two samples. The finding of measurement equivalence across cultures provides essential evidence of construct validity for the schizotypy dimensions and of the cross-cultural validity of SPQ scores. The finding of comparable dimensional structures in cross-cultural samples lends further support to the continuum model of schizotypy and schizophrenia spectrum disorders. Future studies should continue to examine the validity of scores on the SPQ and other schizotypy measures and their variation or consistency across cultures.


Estimating gene signals from noisy microarray images.
In oligonucleotide microarray experiments, noise is a challenging problem, as biologists now are studying their organisms not in isolation but in the context of a natural environment. In low photomultiplier tube (PMT) voltage images, weak gene signals and their interactions with the background fluorescence noise are most problematic. In addition, nonspecific sequences bind to array spots intermittently causing inaccurate measurements. Conventional techniques cannot precisely separate the foreground and the background signals. In this paper, we propose analytically based estimation technique. We assume a priori spot-shape information using a circular outer periphery with an elliptical center hole. We assume Gaussian statistics for modeling both the foreground and background signals. The mean of the foreground signal quantifies the weak gene signal corresponding to the spot, and the variance gives the measure of the undesired binding that causes fluctuation in the measurement. We propose a foreground-signal and shape-estimation algorithm using the Gibbs sampling method. We compare our developed algorithm with the existing Mann-Whitney (MW)- and expectation maximization (EM)/iterated conditional modes (ICM)-based methods. Our method outperforms the existing methods with considerably smaller mean-square error (MSE) for all signal-to-noise ratios (SNRs) in computer-generated images and gives better qualitative results in low-SNR real-data images. Our method is computationally relatively slow because of its inherent sampling operation and hence only applicable to very noisy-spot images. In a realistic example using our method, we show that the gene-signal fluctuations on the estimated foreground are better observed for the input noisy images with relatively higher undesired bindings.


Using a novel T-lymph node ratio model to evaluate the prognosis of nonmetastatic breast cancer patients who received preoperative radiotherapy followed by mastectomy: An observational study.
We aimed to investigate the prognostic value of postpathological characters in nonmetastatic breast cancer (NMBC) patients who received preoperative radiotherapy (PRT) followed by mastectomy (MAST).We conducted retrospective analyses using the data collected from the Surveillance, Epidemiology, and End Results program of the National Cancer Institute. Univariate and multivariate analyses were performed to identify prognostic factors. Disease-specific survival was calculated by the Kaplan-Meier curve and validated by log rank test. The discriminations of independent risk factors and staging systems were compared by the area under receiver operating characteristic curves (AUC) and validated by Harrell concordance index (bootstrapping algorithm). Akaike information criterion (AIC) was applied to compare the difference of model.One thousand three hundred fifty NMBC patients who had received PRT followed by MAST from 1988 to 2013 were included in the study. We found the metastatic lymph node ratio (mLNR) staging was a superior indicator than pN staging. Thus, we proposed a T-lymph node ratio (T-NR) staging system with simplified-T categories (T0-3 and T4) and the mLNR staging. The novel T-NR staging system provided larger AUC (P = .024, .008, respectively) and the smaller AIC (P < .001) value than American Joint Committee on Cancer staging system.The novel T-NR staging system performed more accurate survival prediction and better model fitness for NMBC patients who receive PRT followed by MAST, it may provide a wide applicability in clinical decision-making.


Mitochondrial DNA variation and GIS analysis confirm a secondary origin of geographical variation in the bushcricket Ephippiger ephippiger (Orthoptera: Tettigonioidea), and resurrect two subspecies.
Geographic variation within species can originate through selection and drift in situ (primary variation) or from vicariant episodes (secondary variation). Most patterns of subspecific variation within European flora and fauna are thought to have secondary origins, reflecting isolation in refugia during Quaternary ice ages. The bushcricket Ephippiger ephippiger has an unusual pattern of geographical variability in morphology, behaviour and allozymes in southern France, which has been interpreted as reflecting recent primary origins rather than historical isolation. Re-analysis of this variation using Geographical Information Systems (GIS) suggests a possible zone of hybridization within a complex pattern of geographical variation. Here we produce a genetic distance matrix from restriction fragment length polymorphism (RFLP) bandsharing of an approximately 4.5 kb fragment of mitochondrial DNA (mtDNA), and compare this with predictions resulting from the GIS analysis. The mtDNA variation supports a postglacial origin of geographical variation. Partial Mantel test comparisons of genetic distances with matrices of geographical distance, relevant environmental characteristics and possible refugia show refugia to be the best predictors of genetic distance. There is no evidence to support isolation by distance. However, environmental contrasts do explain significant variation in genetic distance after allowing for the effect of refugial origin. Also, a neighbour-joining tree has a major division separating eastern and western forms. We conclude that the major source of variation within the species is historical isolation in glacial refugia, but that dispersal, hybridization and selection associated with environmental features has influenced patterns of mtDNA introgression. At least two valid subspecies can be defined.


Freedom of racist speech: Ego and expressive threats.
Do claims of "free speech" provide cover for prejudice? We investigate whether this defense of racist or hate speech serves as a justification for prejudice. In a series of 8 studies (N = 1,624), we found that explicit racial prejudice is a reliable predictor of the "free speech defense" of racist expression. Participants endorsed free speech values for singing racists songs or posting racist comments on social media; people high in prejudice endorsed free speech more than people low in prejudice (meta-analytic r = .43). This endorsement was not principled-high levels of prejudice did not predict endorsement of free speech values when identical speech was directed at coworkers or the police. Participants low in explicit racial prejudice actively avoided endorsing free speech values in racialized conditions compared to nonracial conditions, but participants high in racial prejudice increased their endorsement of free speech values in racialized conditions. Three experiments failed to find evidence that defense of racist speech by the highly prejudiced was based in self-relevant or self-protective motives. Two experiments found evidence that the free speech argument protected participants' own freedom to express their attitudes; the defense of other's racist speech seems motivated more by threats to autonomy than threats to self-regard. These studies serve as an elaboration of the Justification-Suppression Model (Crandall & Eshleman, 2003) of prejudice expression. The justification of racist speech by endorsing fundamental political values can serve to buffer racial and hate speech from normative disapproval. (PsycINFO Database Record


Nuclear transfer reprogramming does not improve the low developmental potency of embryonic stem cells induced by long-term culture.
Epigenetic states of embryonic stem (ES) cells are easily altered by long-term cultivation and lose their developmental potential. To rescue this reduced developmental capacity, nuclear transfer (NT) of ES cells was carried out, and original ES and ES cells from cloned blastocysts (ntES) cells established after NT were compared with in vitro differentiation ability and developmental potential by embryoid body formation and tetraploid aggregation respectively. In the establishment of ntES cell lines, the oocytes fused with the ES cell were activated, and further cultured to cloned blastocysts. When in vitro differentiation ability was examined between original and ntES cell lines derived from ES cells with extensive passages (ES-ep), the day of appearance of simple embryoid body, cystic embryoid body, and spontaneous beating was almost similar. The developmental rates of ES-ep cells, that aggregated with tetraploid embryos to term, ranged from 3 to 6%. Moreover, the majority of live pups died soon after birth. In the ntES cell lines derived from ES-ep cells, developmental rates ranged from 0 to 5%. Those pups also died soon after birth, similar to the ES-ep-derived pups. These results suggest that profound epigenetic modifications of ES cells were retained in the re-established cell lines by NT.


Delayed hypersensitivity: bone marrow as the source of cells in delayed skin reactions.
Adult Lewis rats were thymectomized, irradiated, and restored with bone marrow from allogeneic (or F(1)) donors. They were passively sensitized to tuberculin by a subsequent transfer of Lewis lymph node cells and were given intradermal skin tests with tuberculoprotein. In 24-hour skin reactions the majority of cells, in successive experiments, were shown to be allogeneic (or F(1)) with the use of isoantibody against the antigens of the transplanted marrow cells and by the indirect fluorescent antibody technique. Our results demonstrate that the non-specific cells making up a large proportion of the infiltrating elements in tuberculin skin reactions probably originate in the bone marrow.


pp60(v-src) induction of cyclin D1 requires collaborative interactions between the extracellular signal-regulated kinase, p38, and Jun kinase pathways. A role for cAMP response element-binding protein and activating transcription factor-2 in pp60(v-src) signaling in breast cancer cells.
The cyclin D1 gene is overexpressed in breast tumors and encodes a regulatory subunit of cyclin-dependent kinases that phosphorylate the retinoblastoma protein. pp60(c-src) activity is frequently increased in breast tumors; however, the mechanisms governing pp60(c-src) regulation of the cell cycle in breast epithelium are poorly understood. In these studies, pp60(v-src) induced cyclin D1 protein levels and promoter activity (48-fold) in MCF7 cells. Cyclin D1-associated kinase activity and protein levels were increased in mammary tumors from murine mammary tumor virus-pp60(c-src527F) transgenic mice. Optimal induction of cyclin D1 by pp60(v-src) involved the extracellular signal-regulated kinase, p38, and c-Jun N-terminal kinase members of the mitogen-activated protein kinase family. Cyclin D1 promoter activation by pp60(v-src) involved a cAMP response element-binding protein (CREB)/activating transcription factor 2 (ATF-2) binding site. Dominant negative mutants of CREB and ATF-2 but not c-Jun inhibited pp60(v-src) induction of cyclin D1. pp60(v-src) induction of CREB was blocked by the p38 inhibitor SB203580 or by mutation of CREB at Ser133. pp60(v-src) induction of ATF-2 was abolished by the c-Jun N-terminal kinase inhibitor JNK-interacting protein-1 or by mutation of ATF-2 at Thr69 and Thr71. CREB and ATF-2, which bind to a common pp60(v-src) response element, are transcriptionally activated by distinct mitogen-activated protein kinases. Induction of cyclin D1 activity by pp60(v-src) may contribute to breast tumorigenesis through phosphorylation and inactivation of the retinoblastoma protein.


Cholera toxin B subunit promotes the induction of regulatory T cells by preventing human dendritic cell maturation.
Cholera toxin B subunit (CTB) is an efficient mucosal carrier molecule for the generation of immune responses to linked antigens. There is also good evidence that CTB acts as an immunosuppressant, as it is able to down-modulate human monocyte/macrophage cell line activation and to suppress Th1-type responses. In the present study, we examined the possibility that recombinant CTB (rCTB) may affect human dendritic cell (DC) functions in response to LPS stimulation and may induce the generation of DC with the capacity to generate CD4(+) regulatory T cells (Tregs). Our findings show that rCTB partially prevents the LPS-induced maturation process of monocyte-derived DC (MDDC) and decreases their IL-12 production with no relevant effect on IL-10 production. LPS-stimulated MDDC pretreated with rCTB are able to promote the induction of low proliferating T cells, which show an enhanced IL-10 production associated with a reduced IFN-gamma production and the same high levels of TGF-beta as the control. These T cells suppress proliferation of activated autologous T cells. Transwell experiments and blockade of IL-10R and TGF-beta showed that the immunomodulatory effect is mediated by soluble factors. Thus, T cells induced by rCTB-conditioned MDDC acquire a regulatory phenotype and activity similar to those described for type 1 Tregs.


PspA family distribution, antimicrobial resistance and serotype of Streptococcus pneumoniae isolated from upper respiratory tract infections in Japan.
The protection against pneumococcal infections provided by currently available pneumococcal polysaccharide conjugate vaccines are restricted to the limited number of the serotypes included in the vaccine. In the present study, we evaluated the distribution of the pneumococcal capsular type and surface protein A (PspA) family of pneumococcal isolates from upper respiratory tract infections in Japan. A total of 251 S. pneumoniae isolates from patients seeking treatment for upper respiratory tract infections were characterized for PspA family, antibiotic resistance and capsular type. Among the 251 pneumococci studied, the majority (49.4%) was identified as belonging to PspA family 2, while most of the remaining isolates (44.6%) belonged to family 1. There were no significant differences between the distributions of PspA1 versus PspA2 isolates based on the age or gender of the patient, source of the isolates or the isolates' susceptibilities to penicillin G. In contrast, the frequency of the mefA gene presence and of serotypes 15B and 19F were statistically more common among PspA2 strains. The vast majority of pneumococci isolated from the middle ear fluids, nasal discharges/sinus aspirates or pharyngeal secretions represented PspA families 1 and 2. Capsular serotypes were generally not exclusively associated with certain PspA families, although some capsular types showed a much higher proportion of either PspA1 or PspA2. A PspA-containing vaccine would potentially provide high coverage against pneumococcal infectious diseases because it would be cross-protective versus invasive disease with the majority of pneumococci infecting children and adults.


Genomic rearrangements uncovered by genome-wide co-evolution analysis of a major nosocomial pathogen, Enterococcus faecium.
Enterococcus faecium is a gut commensal of the gastro-digestive tract, but also known as nosocomial pathogen among hospitalized patients. Population genetics based on whole-genome sequencing has revealed that E. faecium strains from hospitalized patients form a distinct clade, designated clade A1, and that plasmids are major contributors to the emergence of nosocomial E. faecium. Here we further explored the adaptive evolution of E. faecium using a genome-wide co-evolution study (GWES) to identify co-evolving single-nucleotide polymorphisms (SNPs). We identified three genomic regions harbouring large numbers of SNPs in tight linkage that are not proximal to each other based on the completely assembled chromosome of the clade A1 reference hospital isolate AUS0004. Close examination of these regions revealed that they are located at the borders of four different types of large-scale genomic rearrangements, insertion sites of two different genomic islands and an IS30-like transposon. In non-clade A1 isolates, these regions are adjacent to each other and they lack the insertions of the genomic islands and IS30-like transposon. Additionally, among the clade A1 isolates there is one group of pet isolates lacking the genomic rearrangement and insertion of the genomic islands, suggesting a distinct evolutionary trajectory. In silico analysis of the biological functions of the genes encoded in three regions revealed a common link to a stress response. This suggests that these rearrangements may reflect adaptation to the stringent conditions in the hospital environment, such as antibiotics and detergents, to which bacteria are exposed. In conclusion, to our knowledge, this is the first study using GWES to identify genomic rearrangements, suggesting that there is considerable untapped potential to unravel hidden evolutionary signals from population genomic data.


Recurrent and other new foot ulcers after healed plantar forefoot diabetic ulcer.
Diabetic foot ulcer is a serious complication in patients with diabetes. In most outcome studies of this condition, there is a combination of various types of ulcer and ulcer locations. Plantar ulcers are usually localized to the forefoot, and constitute a quarter of all diabetic foot ulcers. There are a limited number of studies regarding development of new ulcers following healing of a plantar forefoot ulcer, and there are no uniform definitions of recurrent and other new ulcers. The aim of this study was to evaluate the outcome of a large cohort of consecutively treated patients with diabetes mellitus and a healed planter forefoot ulcer (n = 617) with regard to development, characteristics, and outcome of recurrent and other new ulcers. Patients were followed consecutively and prospectively with a 2-year follow-up, according to a preset protocol. Out of 617 patients, 250 (41%) did not develop any new ulcer, 262 (42%) developed a new ulcer, 87 (14%) died and 18 (3%) were lost at 2 years following healing of a plantar forefoot ulcer. Thirty-four percent developed other new ulcers (112 on the same foot and 99 on the contralateral foot), whereas 51 patients (8%) developed a recurrent ulcer (at the same site and foot). Of the patients who died within 2 years, 30 patients had developed other new ulcers. The risk of a recurrent ulcer in patients with diabetes and a healed plantar forefoot ulcer was only 8% within 2 years, whereas other new ulcers, on the same foot or on the contralateral foot, was seen in 4 out of 10 patients indicating the need for further preventive measures and surveillance in these patients. We suggest a concise definition for new ulcer to be used in future research.


Effect of Anisotropy of Potassium Dihydrogen Phosphate Crystals on Its Deformation Mechanisms Subjected to Nanoindentation.
Potassium dihydrogen phosphate (KDP) is an important nonlinear material due to its excellent physical and optical properties. However, it is also a difficult-to-machine material due to its complex anisotropic microstructure. To better understand the deformation mechanisms under external stresses, this paper aims to carry out systematic nanoindentation simulations using molecular dynamics (MD). To facilitate the structural characterization of KDP, a machine learning-based method was developed. The results showed that the subsurface damage is obviously anisotropic. On the (001) surface, both tetragonal and monoclinic phases appear simultaneously and part of the monoclinic phase transfers to the tetragonal phase. The generated phases close to the surface undergo amorphization and are squeezed out to form pile-ups. On the (100) surface, however, an orthorhombic phase emerges directly from the original structure rather than transforming through the monoclinic phase. No amorphization happens and no pile-ups appear in this case. The first "pop-in" in the load-displacement curve of nanoindentation signified the emergence of phase transformation under the combined hydrostatic and shear stresses. After unloading, the recovery of the deformed KDP is also anisotropic. The maximum recovery takes place when the indentation is on the (100) surface.


A systematic, updated review on the antidepressant agomelatine focusing on its melatonergic modulation.
To present an updated, comprehensive review on clinical and pre-clinical studies on agomelatine. A MEDLINE, Psycinfo and Web of Science search (1966-May 2009) was performed using the following keywords: agomelatine, melatonin, S20098, efficacy, safety, adverse effect, pharmacokinetic, pharmacodynamic, major depressive disorder, bipolar disorder, Seasonal Affective Disorder (SAD), Alzheimer, ADHD, Generalized Anxiety Disorder (GAD), Panic Disorder (PD), Obsessive-Compulsive Disorder (OCD), anxiety disorders and mood disorder. STUDY COLLECTION AND DATA EXTRACTION: All articles in English identified by the data sources were evaluated. Randomized, controlled clinical trials involving humans were prioritized in the review. The physiological bases of melatonergic transmission were also examined to deepen the clinical comprehension of agomelatine' melatonergic modulation. Agomelatine, a melatonergic analogue drug acting as MT(1)/MT(2) agonist and 5-HT(2C) antagonist, has been reported to be an effective antidepressant therapy. Although a bias in properly assessing the "sleep core" of depression may still exist with current screening instruments, therefore making difficult to compare agomelatine' efficacy to other antidepressant ones, comparative studies showed agomelatine to be an intriguing option for depression and, potentially, for other therapeutic targets as well.


Assessing the need for a pre-exposure prophylaxis programme using the social media app Grindr®.
HIV pre-exposure prophylaxis (PrEP) is not approved in Switzerland and therefore must be paid for by the users themselves. We conducted a survey to find out whether men who have sex with men (MSM) in Switzerland are already taking PrEP, or are considering using it, and whether it is being taken under medical supervision or not. Grindr® is a geosocial networking app for MSM. Between 5 and 24 January 2017, users of the app who were located in Switzerland by a global positioning system (GPS) were asked to participate in a ten-question survey on PrEP use. Of the 2455 people who took part in the survey, 1893 were included in the analysis. Eighty-two participants (4.3%) reported that they were currently taking PrEP, 64 of whom (78%) said that they were under medical supervision. Seven PrEP users (9%) declared that they had not taken an HIV test within the previous 12 months. Nine hundred and forty-four (49.9%) were considering taking PrEP in the next 6 months, and 1474 (77.9%) were considering taking it at some point in the future. In an online survey carried out among sexually active MSM in Switzerland, only a minority of the individuals approached responded that they were currently using PrEP. However, the majority of participants were considering taking PrEP in the future. We identified a substantial proportion of PrEP users taking PrEP outside a medical setting. Hence, a national programme facilitating access to medical care and providing PrEP is urgently needed.


In vitro DNA replication of recombinant plasmid DNAs containing the origin of progeny replicative form DNA synthesis of phage phi X174.
The origin of phage phi X174 progeny replicative form (RF) DNA synthesis has been inserted into the plasmid vector pBR322 and cloned. In direct contrast to pBR322, the recombinant superhelical plasmids can substitute for phi X174 RFI DNA as template in phi X174-specific reactions in vitro. We have shown that the recombinant plasmids: (i) are cleaved by the phi X174 A protein; (ii) support net synthesis of unit-length single-stranded circular DNA in the presence of the phi X174 A protein and Escherichia coli rep protein, DNA-binding protein, and DNA polymerase III elongation system; (iii) support replication of duplexes catalyzed by the phi X174 A protein and extracts of E. coli.


Extracorporeal photochemotherapy in cutaneous lupus erythematosus.
We report two female subjects with therapy-resistant cutaneous lupus erythematosus (LE), one with subacute cutaneous and the other chronic discoid LE, both treated with extracorporeal photochemotherapy (ECP). The responses after six and eight cycles of ECP led to a prolonged remission of 18 and 11 months, respectively. ECP seems to be a treatment option for patients not responding to or showing unwanted side-effects during conventional standard therapy.


Alamandine attenuates oxidative stress in the right carotid following transverse aortic constriction in mice.
Pressure overload can result in significant changes to the structure of blood vessels, a process known as vascular remodeling. High levels of tension can cause vascular inflammation, fibrosis, and structural alterations to the vascular wall. Prior research from our team has demonstrated that the oral administration of alamandine can promote vasculoprotective effects in mice aorta that have undergone transverse aortic constriction (TAC). Furthermore, changes in local hemodynamics can affect the right and left carotid arteries differently after TAC. Thus, in this study, we aimed to assess the effects of alamandine treatment on right carotid remodeling and the expression of oxidative stress-related substances induced by TAC. Male C57BL/6 mice were categorized into three groups: Sham, TAC, and TAC treated with alamandine (TAC+ALA). Alamandine treatment was administered orally by gavage (30 µg/kg/day), starting three days before the surgery, and continuing for a period of fourteen days. Morphometric analysis of hematoxylin and eosin-stained sections revealed that TAC induced hypertrophic and positive remodeling in the right carotid artery. Picrosirius Red staining also demonstrated an increase in total collagen deposition in the right carotid artery due to TAC-induced vascular changes. Alamandine treatment effectively prevented the increase in reactive oxygen species production and depletion of nitric oxide levels, which were induced by TAC. Finally, alamandine treatment was also shown to prevent the increased expression of nuclear factor erythroid 2-related factor 2 and 3-nitrotyrosine that were induced by TAC. Our results suggest that alamandine can effectively attenuate pathophysiological stress in the right carotid artery of animals subjected to TAC.


Diabetes impairs the angiogenic potential of adipose-derived stem cells by selectively depleting cellular subpopulations.
Pathophysiologic changes associated with diabetes impair new blood vessel formation and wound healing. Mesenchymal stem cells derived from adipose tissue (ASCs) have been used clinically to promote healing, although it remains unclear whether diabetes impairs their functional and therapeutic capacity. In this study, we examined the impact of diabetes on the murine ASC niche as well as on the potential of isolated cells to promote neovascularization in vitro and in vivo. A novel single-cell analytical approach was used to interrogate ASC heterogeneity and subpopulation dynamics in this pathologic setting. Our results demonstrate that diabetes alters the ASC niche in situ and that diabetic ASCs are compromised in their ability to establish a vascular network both in vitro and in vivo. Moreover, these diabetic cells were ineffective in promoting soft tissue neovascularization and wound healing. Single-cell transcriptional analysis identified a subpopulation of cells which was diminished in both type 1 and type 2 models of diabetes. These cells were characterized by the high expression of genes known to be important for new blood vessel growth. Perturbations in specific cellular subpopulations, visible only on a single-cell level, represent a previously unreported mechanism for the dysfunction of diabetic ASCs. These data suggest that the utility of autologous ASCs for cell-based therapies in patients with diabetes may be limited and that interventions to improve cell function before application are warranted.


[Fulminant myocarditis and acute gastroenteritis due to Coxsackie virus B6].
We present the case of a young woman who suffered cardiogenic due to by Coxsackie virus B6. The patient attended a private clinic with an acute gastroenteritis and after one hour of receiving hydratation,she developed hypotension and shock, severe hypoxemia and bilateral lung infiltrate. The patient entered the Intensive Care Unit, where she received hemodynamic support. Due to the clinical picture and cardiac enzymes increase, a cardiac failure was suspected and the echocardiographic findings suggested "myocarditis". The evolution was successful and Coxsackie B6 virus infection diagnosis was made during the follow up by increase of the levels of antibodies for virus Coxsackie B6.


Effect of donor-specific antibodies and panel reactive antibodies in living donor liver transplant recipients.
Preformed circulating donor-specific antibodies (DSAs) immunologically challenge vascular endothelium and the bile duct. However, the liver is an immune-tolerant organ and can avoid immunological challenges. This study was undertaken to analyze the effects of DSAs after adult living donor liver transplantation (LDLT). We retrospectively reviewed 219 LDLT patients' records treated at our center. Of the 219 patients, 32 (14.6%) were DSA (+) and 187 (85.4%) were DSA (-). Class I DSAs were present in 18 patients, class II in seven patients, and both in seven patients. Seven patients (3.2%) showed DSA to HLA-A, four (1.8%) to HLA-B, seven (3.2%) to HLA-DR, and 14 (6.4%) to two or more HLAs. More DSAs were observed in female recipients than male recipients in the DSA (+) group. The DSA (+) group showed significantly higher levels of class I and II panel reactive antibody (PRA) than did the DSA (-) group. No significant intergroup differences were found between incidences of primary nonfunction, acute rejection, vascular complication, or biliary complication. There were no significant differences in graft survival rates between the two groups. However, the recipients with multiple DSAs tended to have more acute rejection episodes and events of biliary stricture and lower graft survival rates than did patients in the DSA (-) group. In LDLT, the presence of multiple DSAs and high PRA seemed to be associated with poor graft outcomes, although our results did not reach statistical significance. Large cohort studies are necessary to clarify the impact of DSA and PRA in LDLT.


Laparoscopic surgery for rectal cancer: The state of the art.
At present time, there is evidence from randomized controlled studies of the success of laparoscopic resection for the treatment of colon cancer with reported smaller incisions, lower morbidity rate and earlier recovery compared to open surgery. Technical limitations and a steep learning curve have limited the wide application of mini-invasive surgery for rectal cancer. The present article discusses the current status of laparoscopic resection for rectal cancer. A review of the more recent retrospective, prospective and randomized controlled trial (RCT) data on laparoscopic resection of rectal cancer including the role of trans-anal endoscopic microsurgery and robotics was performed. A particular emphasis was dedicated to mid and low rectal cancers. Few prospective and RCT trials specifically addressing laparoscopic rectal cancer resection are currently available in the literature. Improved short-term outcomes in term of lesser intraoperative blood loss, reduced analgesic requirements and a shorter hospital stay have been demonstrated. Concerns have recently been raised in the largest RCT trial of the oncological adequacy of laparoscopy in terms of increased rate of circumferential margin. This data however was not confirmed by other prospective comparative studies. Moreover, a similar local recurrence rate has been reported in RCT and comparative series. Similar findings of overall and disease free survival have been reported but the follow-up time period is too short in all these studies and the few RCT trials currently available do not draw any definitive conclusions. On the basis of available data in the literature, the mini-invasive approach to rectal cancer surgery has some short-term advantages and does not seem to confer any disadvantage in term of local recurrence. With respect to long-term survival, a definitive answer cannot be given at present time as the results of RCT trials focused on long-term survival currently ongoing are still to fully clarify this issue.


Attitudes and behaviors of psychiatry residents toward pharmaceutical representatives before and after an educational intervention.
The authors sought to determine the effect of an educational seminar on interactions with pharmaceutical representatives on residents' attitudes and behavior. A controlled trial of an educational intervention was conducted. Residents at a university-affiliated residency program (N=32) were divided into two groups: one group (N=18) received a 1-hour educational intervention, while the other group (N=14) served as a control. Both groups completed a 33-item survey before the intervention and 2 months after the intervention. Residents interacted substantially with pharmaceutical representatives. The majority of residents found the interactions and gifts useful and believed their prescribing practices were not influenced. Compared to the comparison group, the intervention group significantly decreased the reported number of office supplies and noneducational gifts, but showed no change in attitude toward pharmaceutical representatives and their gifts. One-time educational interventions may have significant impact on psychiatric residents' targeted gift-accepting behavior but little effect on attitudes.


Continuous symmetry and chirality measures: approximate algorithms for large molecular structures.
Quantifying imperfect symmetry of molecules can help explore the sources, roles and extent of structural distortion. Based on the established methodology of continuous symmetry and chirality measures, we develop a set of three-dimensional molecular descriptors to estimate distortion of large structures. These three-dimensional geometrical descriptors quantify the gap between the desirable symmetry (or chirality) and the actual one. They are global parameters of the molecular geometry, intuitively defined, and have the ability to detect even minute structural changes of a given molecule across chemistry, including organic, inorganic, and biochemical systems. Application of these methods to large structures is challenging due to countless permutations that are involved in the symmetry operations and have to be accounted for. Our approach focuses on iteratively finding the approximate direction of the symmetry element in the three-dimensional space, and the relevant permutation. Major algorithmic improvements over previous versions are described, showing increased accuracy, reliability and structure preservation. The new algorithms are tested for three sets of molecular structures including pillar[5]arene complexes with Li+, C100 fullerenes, and large unit cells of metal organic frameworks. These developments complement our recent algorithms for calculating continuous symmetry and chirality measures for small molecules as well as protein homomers, and simplify the usage of the full set of measures for various research goals, in molecular modeling, QSAR and cheminformatics.


Head impact exposure in youth football-Are current interventions hitting the target?
Restrictions on heading in youth football have been implemented in some countries to limit head impact exposure. However, current interventions remain poorly guided by evidence. Our objective was to quantify heading exposure in youth football, assessing the effects of sex and age. Football matches played during an international youth football tournament with no heading restrictions were directly observed, including players from both sexes (11-19 years). The elite senior level was included for comparison, using video analysis. All heading events were registered, classified, and assigned to individual players. Heading rates were calculated for each sex and age group. We observed a total of 267 matches, corresponding to 4011 player hours (1927 player hours for females, 2083 player hours for males). Males headed more frequently than females (2.7 vs 1.8 headers/player hour; P < .001). Heading rates increased with age (ANOVA, P < .001), approaching the elite senior level for players 16 years and older. There was substantial variation within teams for all age and sex groups, with the widest range (1-18 headers) observed for girls aged 19. Girls younger than 12 years had the lowest exposure, with an average of <2 players per team heading the ball, each with 1-2 headers. In conclusion, age and sex influence head impact exposure in youth football, and warrants careful consideration when introducing injury prevention measures. Males are more frequently exposed than females, heading rates increase with age, and there is substantial variation between players. Heading is a rare event in the youngest age groups, especially among females.


Identification of NFAT5 as a transcriptional regulator of the EDN1 gene in collecting duct.
The inner medullary collecting duct (IMCD) produces very high levels of endothelin-1 (ET-1) that acts as an autocrine inhibitor of IMCD Na+ and water reabsorption. Recent studies suggest that IMCD ET-1 production is enhanced by extracellular hypertonicity as can occur during high salt intake. Although NFAT5 has been implicated in the IMCD ET-1 hypertonicity response, no studies in any cell type have identified NFAT5 as a transcriptional regulator of the EDN1 gene; the current study examined this using a mouse IMCD cell line (IMCD3). Media hypertonicity increased IMCD3 ET-1 mRNA in a dose- and time-dependent manner associated with increased NFAT5 nuclear localization. Knockdown of NFAT5 using small-interfering RNA or by CRISPR/Cas9-mediated targeting of exon 4 of the NFAT5 gene reduced the ET-1 hypertonicity response. Chromatin immunoprecipitation using an NFAT5 antibody pulled down ET-1 promoter regions containing NFAT5 consensus binding sequences. Transfected ET-1 promoter reporter constructs revealed maximal hypertonicity-induced reporter activity in the proximal 1-kb region; mutation of the two NFAT5 consensus-binding sites in this region abolished hypertonicity-induced reporter activity. The 1-kb ET-1 promoter-reporter construct lost hypertonicity responsiveness when transfected in CRISPR/Cas9-induced NFAT5-deficient cells. In summary, these findings represent the first description that NFAT5 is a direct transcriptional regulator of the EDN1 gene in IMCD cells and point to a potentially important mechanism by which body Na+ homeostasis is maintained.


IMAGING DIAGNOSIS-THE COMPUTED TOMOGRAPHY FEATURES OF A PLEUROPERITONEAL HERNIA IN A CAT.
An 8-year-old female neutered domestic short hair cat presented for investigation of poorly controlled diabetes mellitus. Thoracic radiographs identified a soft tissue opacity in the caudoventral thorax adjacent to the diaphragm. Computed tomography (CT) then characterized a pleuroperitoneal hernia with cranial displacement of a portion of the liver within the hernia. A pleuroperitoneal hernia was confirmed and repaired via exploratory laparotomy. This is the first description of the CT features of a pleuroperitoneal hernia in a cat.


New Insights into the Role of Inflammation in the Pathogenesis of Atherosclerosis.
Atherosclerosis is a chronic inflammatory disease characterized by the accumulation of lipids, smooth muscle cell proliferation, cell apoptosis, necrosis, fibrosis, and local inflammation. Immune and inflammatory responses have significant effects on every phase of atherosclerosis, and increasing evidence shows that immunity plays a more important role in atherosclerosis by tightly regulating its progression. Therefore, understanding the relationship between immune responses and the atherosclerotic microenvironment is extremely important. This article reviews existing knowledge regarding the pathogenesis of immune responses in the atherosclerotic microenvironment, and the immune mechanisms involved in atherosclerosis formation and activation.


HSV cheats the executioner.
In this issue of Cell Host & Microbe, Huang et al. (2015) and Guo et al. (2015), along with recent work by Wang et al. (2014), reveal that HSV ribonucleotide reductase has opposing activities in either inducing or preventing necroptosis, depending on the host species. This evolutionary twist underscores the importance of selective pressure in virus-host relationships.


Differences in Trochlear Surface Damage and Wear Between Three Different Total Knee Arthroplasty Designs.
Trochlear design plays a role in patellofemoral kinematics. Little is known regarding differences in the contact properties between modern designs. The purpose of the present study was to analyze patellofemoral joint contact by identifying areas of joint surface damage and wear in retrieved femoral components of 3 modern designs. Eighteen retrieved femoral components featuring 3 different modern designs (Triathlon, SIGMA, and GENESIS II) were matched based on time-in-vivo, age, gender, and body mass index. Trochlear wear and surface damage were assessed using visual inspection, light microscopy, and light profilometry. Visual inspection of the femoral components showed evidence of surface damage in all implant types. No significant differences between the groups were found with respect to surface damage and wear on visual inspection. Light profilometry of retrieved components showed that retrieved Triathlon (P = .002) and SIGMA (P = .009) components were significantly rougher than the corresponding reference components. The GENESIS II retrieved components were not substantially rougher than the reference components (P = .48). Light profilometry analysis showed significantly increased roughness of retrieved SIGMA components compared with Triathlon or GENESIS II components, particularly in the proximal trochlear groove and the medial trochlea. As the volume and patient demands for total knee arthroplasty increase, a greater understanding of the patellofemoral joint is warranted. Patellofemoral kinematics can have an effect on the surface characteristics of total knee arthroplasty components. Retrieved SIGMA components appeared to have increased roughness compared with Triathlon or GENESIS II components. The etiology and long-term effects of increased trochlear roughness require further investigation.


Class 1 integron-associated gene cassettes in Salmonella enterica subsp. enterica serovar Agona isolated from pig carcasses in Brazil.
Two multiresistant Salmonella enterica subsp. enterica serovar Agona isolates from pig carcasses were investigated for antimicrobial resistance genes and their location with particular reference to the detection of class 1 integrons. The two S. Agona isolates were investigated for their in vitro susceptibility to antimicrobial agents and their plasmid content. The resistance genes and class 1 amplicons were identified by PCR assays. Amplicons of class 1 integrons were cloned and sequenced. Transferability of resistance plasmids was confirmed by conjugation. Both S. Agona isolates carried conjugative plasmids of approximately 150 kb which harboured all resistance genes detected in the respective isolates. S. Agona 231 was resistant to chloramphenicol by catA1, to tetracycline and minocycline by tet(B), and to sulphonamides by sul1. In addition, it harboured a streptomycin resistance gene strA and a class 1 integron with a new aadA variant designated aadA23, which mediates resistance to streptomycin and spectinomycin. S. Agona 242 also carried the genes catA1, tet(B), and sul1. Moreover, it harboured a second sulphonamide resistance gene, sul2, and a class 1 integron with intact gene cassettes carrying new variants of the trimethoprim resistance gene dfrA15b or the chloramphenicol resistance gene cmlA4. The third gene cassette consisted of a truncated aadA2 gene. The results of this study show that large conjugative multiresistance plasmids are present in S. Agona from pigs. Analysis of the class 1 integrons revealed the presence of new variants of resistance genes so far not detected in Salmonella isolates.


Influence of DNMT genotype on global and site specific DNA methylation patterns in neonates and pregnant women.
This study examines the relationship between common genetic variation within DNA methyltransferase genes and inter-individual variation in DNA methylation. Eleven polymorphisms spanning DNMT1 and DNMT3B were genotyped. Global and gene specific (IGF2, IGFBP3, ZNT5) DNA methylation was quantified by LUMA and bisulfite Pyrosequencing assays, respectively, in neonatal cord blood and in maternal peripheral blood. Associations between maternal genotype and maternal methylation (n (≈) 333), neonatal genotype and neonatal methylation (n (≈) 454), and maternal genotype and neonatal methylation (n (≈) 137) were assessed. The findings of this study provide some support to the hypothesis that genetic variation in DNA methylating enzymes influence DNA methylation at global and gene-specific levels; however observations were not robust to correction for multiple testing. More comprehensive analysis of the influence of genetic variation on global and site specific DNA methylation is warranted.


Fully automated identification of brain abnormality from whole-body FDG-PET imaging using deep learning-based brain extraction and statistical parametric mapping.
The whole brain is often covered in [18F]Fluorodeoxyglucose positron emission tomography ([18F]FDG-PET) in oncology patients, but the covered brain abnormality is typically screened by visual interpretation without quantitative analysis in clinical practice. In this study, we aimed to develop a fully automated quantitative interpretation pipeline of brain volume from an oncology PET image. We retrospectively collected 500 oncologic [18F]FDG-PET scans for training and validation of the automated brain extractor. We trained the model for extracting brain volume with two manually drawn bounding boxes on maximal intensity projection images. ResNet-50, a 2-D convolutional neural network (CNN), was used for the model training. The brain volume was automatically extracted using the CNN model and spatially normalized. For validation of the trained model and an application of this automated analytic method, we enrolled 24 subjects with small cell lung cancer (SCLC) and performed voxel-wise two-sample T test for automatic detection of metastatic lesions. The deep learning-based brain extractor successfully identified the existence of whole-brain volume, with an accuracy of 98% for the validation set. The performance of extracting the brain measured by the intersection-over-union of 3-D bounding boxes was 72.9 ± 12.5% for the validation set. As an example of the application to automatically identify brain abnormality, this approach successfully identified the metastatic lesions in three of the four cases of SCLC patients with brain metastasis. Based on the deep learning-based model, extraction of the brain volume from whole-body PET was successfully performed. We suggest this fully automated approach could be used for the quantitative analysis of brain metabolic patterns to identify abnormalities during clinical interpretation of oncologic PET studies.


Delivery of an enzyme-IGFII fusion protein to the mouse brain is therapeutic for mucopolysaccharidosis type IIIB.
Mucopolysaccharidosis type IIIB (MPS IIIB, Sanfilippo syndrome type B) is a lysosomal storage disease characterized by profound intellectual disability, dementia, and a lifespan of about two decades. The cause is mutation in the gene encoding α-N-acetylglucosaminidase (NAGLU), deficiency of NAGLU, and accumulation of heparan sulfate. Impediments to enzyme replacement therapy are the absence of mannose 6-phosphate on recombinant human NAGLU and the blood-brain barrier. To overcome the first impediment, a fusion protein of recombinant NAGLU and a fragment of insulin-like growth factor II (IGFII) was prepared for endocytosis by the mannose 6-phosphate/IGFII receptor. To bypass the blood-brain barrier, the fusion protein ("enzyme") in artificial cerebrospinal fluid ("vehicle") was administered intracerebroventricularly to the brain of adult MPS IIIB mice, four times over 2 wk. The brains were analyzed 1-28 d later and compared with brains of MPS IIIB mice that received vehicle alone or control (heterozygous) mice that received vehicle. There was marked uptake of the administered enzyme in many parts of the brain, where it persisted with a half-life of approximately 10 d. Heparan sulfate, and especially disease-specific heparan sulfate, was reduced to control level. A number of secondary accumulations in neurons [β-hexosaminidase, LAMP1(lysosome-associated membrane protein 1), SCMAS (subunit c of mitochondrial ATP synthase), glypican 5, β-amyloid, P-tau] were reduced almost to control level. CD68, a microglial protein, was reduced halfway. A large amount of enzyme also appeared in liver cells, where it reduced heparan sulfate and β-hexosaminidase accumulation to control levels. These results suggest the feasibility of enzyme replacement therapy for MPS IIIB.


Interactive rhythms across species: the evolutionary biology of animal chorusing and turn-taking.
The study of human language is progressively moving toward comparative and interactive frameworks, extending the concept of turn-taking to animal communication. While such an endeavor will help us understand the interactive origins of language, any theoretical account for cross-species turn-taking should consider three key points. First, animal turn-taking must incorporate biological studies on animal chorusing, namely how different species coordinate their signals over time. Second, while concepts employed in human communication and turn-taking, such as intentionality, are still debated in animal behavior, lower level mechanisms with clear neurobiological bases can explain much of animal interactive behavior. Third, social behavior, interactivity, and cooperation can be orthogonal, and the alternation of animal signals need not be cooperative. Considering turn-taking a subset of chorusing in the rhythmic dimension may avoid overinterpretation and enhance the comparability of future empirical work.


JAK2 V617F impairs hematopoietic stem cell function in a conditional knock-in mouse model of JAK2 V617F-positive essential thrombocythemia.
The JAK2 V617F mutation is found in most patients with a myeloproliferative neoplasm and is sufficient to produce a myeloproliferative phenotype in murine retroviral transplantation or transgenic models. However, several lines of evidence suggest that disease phenotype is influenced by the level of mutant JAK2 signaling, and we have therefore generated a conditional knock-in mouse in which a human JAK2 V617F is expressed under the control of the mouse Jak2 locus. Human and murine Jak2 transcripts are expressed at similar levels, and mice develop modest increases in hemoglobin and platelet levels reminiscent of human JAK2 V617F-positive essential thrombocythemia. The phenotype is transplantable and accompanied by increased terminal erythroid and megakaryocyte differentiation together with increased numbers of clonogenic progenitors, including erythropoietin-independent erythroid colonies. Unexpectedly, JAK2(V617F) mice develop reduced numbers of lineage(-)Sca-1(+)c-Kit(+) cells, which exhibit increased DNA damage, reduced apoptosis, and reduced cell cycling. Moreover, competitive bone marrow transplantation studies demonstrated impaired hematopoietic stem cell function in JAK2(V617F) mice. These results suggest that the chronicity of human myeloproliferative neoplasms may reflect a balance between impaired hematopoietic stem cell function and the accumulation of additional mutations.


Cultivation of hepatitis C virus in primary hepatocyte culture from patients with chronic hepatitis C results in release of high titre infectious virus.
To investigate the viral replication cycle and genomic heterogeneity of hepatitis C virus (HCV), we established an HCV cultivation system by using a primary hepatocyte culture from patients with chronic hepatitis C. Liver tissue was obtained by needle biopsy or surgery, then hepatocytes were isolated by collagenase digestion. After several weeks, we determined the HCV RNA titre of the cultured cells and supernatant by a competitive polymerase chain reaction (PCR) method. A significant amount of HCV RNA was observed in the cells and supernatant during cultivation. Negative-strand RNA, regarded as a marker of viral replication, could be detected by a strand-specific reverse transcription PCR method and the HCV core protein could be detected by immunofluorescence microscopy. Many HCV particles released into the supernatant were infectious. In addition, we compared the nucleotide sequences in the E2/NS1 region of pre- and post-cultivation hepatocytes for 8 weeks. At the beginning of the culture period, three major HCV types containing two subtypes were isolated. Following cultivation, the same types were isolated from the cultured hepatocytes in the same ratio as prior to cultivation. We could detect the same clones in this patient's serum, but in vivo we observed genetic variability over a 6 month interval. One clone detected throughout the 6 month period mutated extensively in the hypervariable region. These results indicated that HCV can replicate in cultured hepatocytes, and that infectious virions are released into the supernatant. This cultivation system should facilitate the study of HCV genomic heterogeneity, infection and replication.


Does interleukin-1 save B cells from paralysis? Studies with pre-B cells and antigen-reactive B cells.
The role of the second messenger cAMP in the differentiation of the pre-B cell line 70Z/3 and in the initial phase of the immune response of B cells, IgM antibody production is examined. Our data indicate that in these B cells, the elevation of intracellular cAMP levels can have two fundamentally different effects: it may dedifferentiate B cells and cause their disintegration (catabolic pathway) or it may induce their differentiation and maturation (anabolic pathway). The switch that determines which pathway B cells enter is set by interleukin-1. Our experiments indicate that intracellular cAMP levels rise in B cells when they interact with helper T cells in a cognate interaction. Given the dual effect that cAMP may have on the fate of B cells, activation or destruction, our data may be taken to suggest that helper T cells may direct B cells into catabolic or anabolic responses, depending on the availability of a costimulatory signal mediated by interleukin-1.


Brain volumes, RBC status, and hepatic function in alcoholics after 1 and 4 weeks of sobriety: predictors of outcome.
The authors asked if hematological indices of RBC status and hepatic function in newly sober alcoholic men are related to abnormalities in brain morphology, change with normalization of brain function during short-term sobriety, and predict prolonged sobriety. Alcoholic men received brain magnetic resonance imaging and laboratory assessments on admission and before discharge from an inpatient treatment program. Healthy comparison men were similarly tested. On admission, RBC count, hemoglobin level, and hematocrit were significantly lower in alcoholic subjects than comparison subjects; mean corpuscular volume, SGOT, SGPT, and gamma-glutamyl transpeptidase were significantly higher. By discharge, all measures had improved, although RBC count, mean corpuscular volume, and gamma-glutamyl transpeptidase levels remained significantly different from those of comparison subjects. Upon admission, alcoholic men had smaller cortical white and gray matter and larger lateral and third ventricle volumes, with reduced lateral ventricle and increased anterior cortical gray matter volumes by discharge. Lower RBC count, hemoglobin level, and hematocrit were associated with lower white matter and higher ventricular volumes at admission. Change in these measures was related to reduction in ventricular volume with treatment. By discharge, associations among RBC count, hemoglobin level, and hematocrit and white matter and ventricular volumes were less marked than at admission. Discharge hemoglobin value and hematocrit discriminated patients who maintained sobriety from those who relapsed. Hepatic function showed limited association with brain measures at admission and discharge. Hemograms reflect alcohol-related abnormalities in brain morphology, improvement over short-term sobriety, and liability to relapse after treatment.


Anatomic Coracoclavicular Ligament Reconstruction for the Treatment of Acute Acromioclavicular Joint Dislocation: Minimum 10-Year Follow-up.
The long-term clinical and radiographic outcomes following coracoclavicular (CC) ligament reconstruction for the operative treatment of acute acromioclavicular (AC) joint dislocation remain uncertain. The purpose of the present study was to determine the long-term clinical and radiographic outcomes of CC ligament reconstruction and to identify risk factors for unfavorable outcomes. We reviewed 20 cases of AC joint dislocation in 19 patients (18 male and 1 female; mean age, 32.3 years) that were treated with single-bundle reconstruction. The mean duration of follow-up was 12.7 years. We measured the CC vertical distance (CCD) on the anteroposterior view and compared the affected and unaffected sides (CCD ratio). We divided the patients into those with a CCD ratio of <25% (Group 1) and those with a CCD ratio of ≥25% (Group 2). We radiographically investigated the clavicular tunnel anteroposterior (CTAP) angle, clavicular tunnel ratio, and coracoid tunnel orientation on the basis of the entry and exit points at the base of the coracoid. For the coracoid tunnel orientation, we compared center-center orientation and noncenter-center orientation. Group 1 comprised 17 cases (85%), and Group 2 comprised 3 cases (15%). At the time of the latest follow-up, Group 1 had a significantly higher mean Constant score than Group 2 (98.2 compared with 90.7; p = 0.038). Of the 3 radiographic parameters, only the CTAP angle was significantly different between the 2 groups (p < 0.0001). Two (67%) of the 3 cases in Group 2 were associated with posterior AC joint displacement. CC ligament reconstruction for the treatment of acute AC joint dislocation resulted in successful long-term clinical and radiographic outcomes. It is important to decrease the CTAP angle and to ensure proper anatomic placement of the clavicular and coracoid tunnels at the time of surgery. Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.


Lipid biomarker and phylogenetic analyses to reveal archaeal biodiversity and distribution in hypersaline microbial mat and underlying sediment.
This study has utilized the tools of lipid biomarker chemistry and molecular phylogenetic analyses to assess the archaeal contribution to diversity and abundance within a microbial mat and underlying sediment from a hypersaline lagoon in Baja California. Based on abundance of ether-linked isoprenoids, archaea made up from 1 to 4% of the cell numbers throughout the upper 100 mm of mat and sediment core. Below this depth archaeal lipid was two times more abundant than bacterial. Archaeol was the primary archaeal lipid in all layers. Relatively small amounts of caldarchaeol (dibiphytanyl glyceroltetraether) were present at most depths with phytanyl to biphytanyl molar ratios lowest (approximately 10 : 1) in the 4-17 mm and 100-130 mm horizons, and highest (132 : 1) in the surface 0-2 mm. Lipids with cyclic biphytanyl cores were only detected below 100 mm. A novel polar lipid containing a C(30) isoprenoid (squalane) moiety was isolated from the upper anoxic portion of the core and partially characterized. Hydrocarbon biomarker lipids included pentamethylicosane (2-10 mm) and crocetane (primarily below 10 mm). Archaeal molecular diversity varied somewhat with depth. With the exception of samples at 0-2 mm and 35-65 mm, Thermoplasmatales of marine benthic group D dominated clone libraries. A significant number of phylotypes representing the Crenarchaeota from marine benthic group B were generally present below 17 mm and dominated the 35-65 mm sample. Halobacteriaceae family made up 80% of the clone library of the surface 2 mm, and consisted primarily of sequences affiliated with the haloalkaliphilic Natronomonas pharaonis.


Evolution of developmental cyclic adenosine monophosphate signaling in the Dictyostelia from an amoebozoan stress response.
The Dictyostelid social amoebas represent one of nature's several inventions of multicellularity. Though normally feeding as single cells, nutrient stress triggers the collection of amoebas into colonies that form delicately shaped fruiting structures in which the cells differentiate into spores and up to three cell types to support the spore mass. Cyclic adenosine monophosphate (cAMP) plays a very dominant role in controlling morphogenesis and cell differentiation in the model species Dictyostelium discoideum. As a secreted chemoattractant cAMP coordinates cell movement during aggregation and fruiting body morphogenesis. Secreted cAMP also controls gene expression at different developmental stages, while intracellular cAMP is extensively used to transduce the effect of other stimuli that control the developmental program. In this review, I present an overview of the different roles of cAMP in the model D. discoideum and I summarize studies aimed to resolve how these roles emerged during Dictyostelid evolution.


The impact of genomic context on outcomes of solid cancer patients treated with genotype-matched targeted therapies: a comprehensive review.
A critical need in the field of genotype-matched targeted therapy in cancer is to identify patients unlikely to respond to precision medicines. This will manage expectations of individualised therapies and avoid clinical progression to a point where institution of alternative treatments might not be possible. We examined the evidence base of the impact of genomic context on which targeted alterations are inscribed to identify baseline biomarkers distinguishing those obtaining the expected response from those with less benefit from targeted therapies. A comprehensive narrative review was conducted: scoping searches were undertaken in PubMed, Cochrane Database of Systematic Reviews, and PROSPERO. Outcomes included in meta-analysis were progression-free and overall survival. Data were extracted from Kaplan-Meier and used to calculate hazard ratios. Studies presenting data on two molecular subcohorts (e.g. co-mutation versus no co-mutation) were included in fixed meta-analysis. Other studies were used for descriptive purposes. The presence of concomitant driver mutations, higher tumour mutational burden (TMB), greater copy number burden, and APOBEC signatures significantly reduces benefits of targeted therapy in lung cancers in never smokers (LCINS - less than 100 cigarettes per lifetime) and breast cancer, cancers with low TMB. LCINS have significantly poorer outcomes if their cancers harbour p53 co-mutations, an effect also seen in human epidermal growth factor receptor 2-positive (HER2+) breast cancer patients (trastuzumab) and head and neck cancer patients [phosphoinositide 3-kinase (PI3K) inhibition]. PI3K co-alterations have less impact when targeting epidermal growth factor receptor mutations and anaplastic lymphoma kinase fusions, but significantly reduce the impact of targeting HER2 and MET amplifications. SMARCA4 co-mutations predict for poor outcome in patients treated with osimertinib and sotorasib. In BRAF-mutant melanoma, whilst there are no genomic features distinguishing exceptional responders from primary progressors, there are clear transcriptomic features dichotomising these outcomes. To our knowledge, this is the most comprehensive review to date of the impact of genomic context on outcomes with targeted therapy. It represents a valuable resource informing progress towards contextualised precision medicine.


Dual-Energy Computed Tomography of the Knee, Ankle, and Foot: Noninvasive Diagnosis of Gout and Quantification of Monosodium Urate in Tendons and Ligaments.
Gout is a true crystal deposition arthropathy caused by the precipitation of monosodium urate into joints and periarticular soft tissues. It is the most common inflammatory arthropathy in men and women of older age with a male-to-female ratio of 3 to 8:1. The disease may progress from asymptomatic hyperuricemia through symptomatic acute gout attacks with asymptomatic periods into chronic symptomatic tophaceous gout. Although invasive arthrocentesis and demonstration of monosodium urate crystals on polarized light microscopy is definitive for the diagnosis of gout, dual-energy computed tomography (CT) allows for noninvasive visualization and reproducible volume quantification of monosodium urate crystals. Based on the high diagnostic performance, dual-energy CT has been included in the 2015 American College of Rheumatology/European League Against Rheumatism Collaborative Initiative Classification Criteria for Gout. Increasing evidence indicates the usefulness of dual-energy CT to guide the management of patients with suspected gout and monitor the effectiveness of urate-lowering medical therapy.


A new scheme for serotyping group-G streptococci.
Ten antisera, prepared against nine selected strains of group-G streptococci and a strain of group-A streptococci of M-type 12, were used to serotype 102 isolates of group-G streptococci by means of precipitation reactions between the sera and hot-acid extracts of the streptococci. Fifty-six (54.9%) of the streptococci could be typed; eight serotypes were identified, of which type VIII was the most common (18.6%). The M12 and the R28 antigens, previously recorded in group-G streptococci, were not detected. The type antigens were trypsin sensitive and resembled the M antigens of group-A streptococci.


Effects of soybean isoflavones on Wnt/β-catenin and the TGF-β1 signaling pathway in renal tissue of type 2 diabetic rats.
To observe the effect of Soyisoflavones (SI) on the expression of Wnt/β-catenin signaling pathway elements, transforming growth factor-β (THGF-β) and its related proteins in the renal interstitia of diabetic nephropathic (DN) rats, 48 DN rats were randomly divided into 4 groups: DN model group (group DN), soybean isoflavone treatment group (group DA), DN model group + losartan treatment group (group DL), DN model group + soybean isoflavones combined with losartan treatment group (group SL). Each group comprised 12 rats. Twelve healthy Wistar rats were selected as normal controls (group N). After 12 weeks of continuous administration of soybean isoflavone or losartan or those two combined, the body weight of rats was recorded and serum urea nitrogen (BUN) and creatinine (Scr) were measured. The expression of Wnt4, β-catenin, and TGF-β1 proteins, as well as mRNA, in the renal interstitium were detected by immunohistochemistry and real-time quantitative PCR (FQ-PCR). In all the groups, Wnt4, β-catenin and TGF-β1 protein were only expressed in renal interstitial and renal tubular epithelial cells. There was no significant difference between group DA and group DL (P>0.05). FQ-PCR results showed that Wnt4, β-catenin and TGF-β1 mRNA were consistent with the expression of these proteins in the renal tissue of each group. Soy isoflavones can reduce 24-h urinary protein quantification, alleviate renal interstitial pathological damage, and regulate the expression of Wnt4, β-catenin and TGF-β1 in the renal interstitium. This suggests that soybean isoflavones could delay the process of renal interstitial fibrosis in DN rats by decreasing the expression of Wnt4, β-catenin and TGF-β1 in the renal interstitium, thus demonstrating that soybean isoflavones plus losartan have the best protective effects against diabetes-induced renal fibrosis.


Identification of Spinal Inhibitory Interneurons Required for Attenuating Effect of Duloxetine on Neuropathic Allodynia-like Signs in Rats.
Neuropathic pain is a chronic pain condition that occurs after nerve damage; allodynia, which refers to pain caused by generally innocuous stimuli, is a hallmark symptom. Although allodynia is often resistant to analgesics, the antidepressant duloxetine has been used as an effective therapeutic option. Duloxetine increases spinal noradrenaline (NA) levels by inhibiting its transporter at NAergic terminals in the spinal dorsal horn (SDH), which has been proposed to contribute to its pain-relieving effect. However, the mechanism through which duloxetine suppresses neuropathic allodynia remains unclear. Here, we identified an SDH inhibitory interneuron subset (captured by adeno-associated viral (AAV) vectors incorporating a rat neuropeptide Y promoter; AAV-NpyP+ neurons) that is mostly depolarized by NA. Furthermore, this excitatory effect was suppressed by pharmacological blockade or genetic knockdown of α1B-adrenoceptors (ARs) in AAV-NpyP+ SDH neurons. We found that duloxetine suppressed Aβ fiber-mediated allodynia-like behavioral responses after nerve injury and that this effect was not observed in AAV-NpyP+ SDH neuron-selective α1B-AR-knockdown. These results indicate that α1B-AR and AAV-NpyP+ neurons are critical targets for spinal NA and are necessary for the therapeutic effect of duloxetine on neuropathic pain, which can support the development of novel analgesics.


An investigation of the antigenic determinants on chloroperoxidase and purified rat liver microsomal cytochrome P-450b.
Chloroperoxidase (CPO) exhibits many physicochemical and catalytic properties similar to those of the bacterial and microsomal cytochromes P-450. Therefore, the possible similarities between the antigenic determinants of CPO and rat liver microsomal cytochrome P-450b were investigated. Polyclonal antibodies against CPO and rat liver cytochrome P-450b were raised in rabbits and used to investigate the antigenic cross-reactivity between CPO and P-450b. Although anti-CPO antibodies were capable of inhibiting the ethyl hydroperoxide-supported N,N-dimethylaniline (DMA) demethylation activity of CPO by more than 80%, they were unable to inhibit the NADPH-supported demethylation of DMA by cytochrome P-450b in the reconstituted system. The ethyl hydroperoxide-supported demethylation of DMA by CPO was not affected by the addition of anti-P-450b antibodies which inhibited cytochrome P-450 activity greater than 90%. In order to probe for the possible existence of common antigenic determinants which were not involved in catalytic activity, the cross-reactivities were investigated using enzyme-linked immunosorbent assays. There was no cross-reactivity between anti-CPO and cytochrome P-450b, or anti-P-450b and CPO using enzyme-linked immunosorbent assays. When control, phenobarbital-, isosafrole-, and beta-naphthoflavone-induced rat and rabbit liver microsomes and CPO were analyzed by Western blotting and developed with anti-P-450 antibodies, only the phenobarbital- and isosafrole-induced microsomes showed a positive reaction in the P-450 region. When anti-CPO antibodies were used on Western blots of the same series of proteins, a positive reaction was observed only with CPO.(ABSTRACT TRUNCATED AT 250 WORDS)


Antibacterial activity and bonding characteristics of an adhesive resin containing antibacterial monomer MDPB.
The aims of this study were to investigate the antibacterial activity of an adhesive resin incorporating the antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) as well as its bonding characteristics in terms of bond strength into dentin and curing ability. An experimental adhesive resin was prepared by incorporation of 2.5% MDPB into proprietary adhesive (LB Bond), and the inhibitory effect of cured specimen against the growth of Streptococcus mutans on its surface was determined. Bond strength to human dentin and degree of conversion of the experimental adhesive in combination with 1, 2, or 5% MDPB-containing primers, which were previously reported to possess bactericidal effect, were evaluated by conventional tensile bond strength test and Fourier transformation infrared spectroscopy. The cured experimental adhesive exhibited an inhibitory effect on S. mutans growth, reducing the number of bacteria to approximately 3% of control adhesive without MDPB. Tensile bond strength of experimental adhesive was not significantly different from that of the control (p>0.05), and use in combination with MDPB-containing primer did not show any adverse influence on bond strength. The degree of conversion of the bonding system was not different (p>0.05) between control and experimental adhesive irrespective of addition of MDPB into primer. These results indicate that an adhesive resin with antibacterial activity after curing could be produced by incorporation of MDPB without influencing bond strength or curing performance, suggesting that the comprehensive bonding system including MDPB-containing primer and adhesive should be highly effective in its antibacterial effect before and after curing.


Adherence to the Mediterranean diet and fresh fruit intake are associated with improved asthma control.
The traditional Mediterranean diet is claimed to possess antioxidant and immune-regulatory properties in several chronic diseases. Typical Mediterranean foods have recently been associated with improvement of symptoms of asthma and rhinitis in children. However the effect of adherence to Mediterranean diet on adult asthma outcomes is unknown. We aimed to investigate the association between adherence to Mediterranean diet and asthma control. Cross sectional study of 174 asthmatics, mean (SD) age of 40 (15) years. The patients were defined as controlled, in contrast to noncontrolled, if they showed FEV1 >or= 80% of predicted, exhaled nitric oxide (NO) <or=35 ppb, and Asthma Control Questionnaire score <1. Dietary intake was obtained by a food frequency questionnaire, and Mediterranean diet was assessed by alternate Mediterranean Diet (aMED) Score. Logistic regression models adjusting for confounders were performed to estimate the association between Mediterranean diet and asthma control. Controlled asthmatics (23%) had significantly higher aMED Score, intake of fresh fruit, and lower intake of ethanol compared to noncontrolled (77%). High adherence to Mediterranean diet reduced 78% the risk of noncontrolled asthma after adjusting for gender, age, education, inhaled corticosteroids and energy intake (OR = 0.22; 95% CI = 0.05-0.85; P-trend = 0.028). The higher intake of fresh fruit decreased the probability of having noncontrolled asthma (OR = 0.29; 95% CI = 0.10-0.83; P-trend = 0.015), while the higher intake of ethanol had the opposite effect (OR = 3.16; 95% CI = 1.10-9.11; P-trend = 0.035). High adherence to traditional Mediterranean diet increased the likelihood of asthma to be under control in adults. The study introduces a novel link between diet and asthma control, as measured by symptoms, lung function and exhaled NO.


Candidate Genes for Inherited Autism Susceptibility in the Lebanese Population.
Autism spectrum disorder (ASD) is characterized by ritualistic-repetitive behaviors and impaired verbal/non-verbal communication. Many ASD susceptibility genes implicated in neuronal pathways/brain development have been identified. The Lebanese population is ideal for uncovering recessive genes because of shared ancestry and a high rate of consanguineous marriages. Aims here are to analyze for published ASD genes and uncover novel inherited ASD susceptibility genes specific to the Lebanese. We recruited 36 ASD families (ASD: 37, unaffected parents: 36, unaffected siblings: 33) and 100 unaffected Lebanese controls. Cytogenetics 2.7 M Microarrays/CytoScan™ HD arrays allowed mapping of homozygous regions of the genome. The CNTNAP2 gene was screened by Sanger sequencing. Homozygosity mapping uncovered DPP4, TRHR, and MLF1 as novel candidate susceptibility genes for ASD in the Lebanese. Sequencing of hot spot exons in CNTNAP2 led to discovery of a 5 bp insertion in 23/37 ASD patients. This mutation was present in unaffected family members and unaffected Lebanese controls. Although a slight increase in number was observed in ASD patients and family members compared to controls, there were no significant differences in allele frequencies between affecteds and controls (C/TTCTG: γ2 value = 0.014; p = 0.904). The CNTNAP2 polymorphism identified in this population, hence, is not linked to the ASD phenotype.


Health may be compromised by social interactions depending on culture among postpartum Arab and Jewish Israeli women.
Social support is generally perceived to facilitate health in postpartum women; however, previous research shows that this is not always true. Social interactions intended to provide support can be perceived as negative and in turn, may have negative impacts on maternal health. The purpose of the present study was to asses if social support and negative interactions at one month after childbirth can predict maternal health four months after childbirth, and if this relationship is influenced by culture. This prospective longitudinal cohort study included randomly selected Arab (n = 203) and Jewish (n = 202) women who attended Mother and Child Health Clinics in Northern Israel one month after giving birth. The women were interviewed at one and four months after childbirth using a questionnaire including measures of health (self-reported health (SRH) and health problems), socioeconomic and demographic status, obstetric characteristics, social support, negative social interactions and perceptions of customs and traditions intended to help the mother cope after childbirth. Multivariable regressions were run to identify the variables predicting health four months after childbirth. The response rate for both interviews was 90%. Negative social interactions one month after childbirth significantly predicted health problems in Arab and Jewish women (Beta 0.20 and 0.37 respectively) and SRH among Arab women only (odds ratio (OR) 0.32, confidence interval (CI) 0.19-0.54) four months after childbirth. Social support at one month after childbirth significantly predicted better SRH in both Jewish and Arab women four months after childbirth (OR 2.33, CI 1.38-3.93 and 1.59, CI 1.01-2.46 respectively) and fewer health problems only among Jewish women (Beta - 0.37). Social support and negative social interactions appear to be predictive of health in postpartum women. Associations varied between Arabs and Jews, indicating that social support may be more important for predicting health among Jewish women and negative interactions may be more important among Arab women. Healthcare practitioners should be aware of the cultural context and social circumstances of postpartum women to ensure they receive the social support and care they need.


Development of a novel risk score to predict mortality in patients admitted to hospital with COVID-19.
Patients hospitalised with COVID-19 have a high mortality. Identification of patients at increased risk of adverse outcome would be important, to allow closer observation and earlier medical intervention for those at risk, and to objectively guide prognosis for friends and family of affected individuals. We conducted a single-centre retrospective cohort study in all-comers with COVID-19 admitted to a large general hospital in the United Kingdom. Clinical characteristics and features on admission, including observations, haematological and biochemical characteristics, were used to develop a score to predict 30-day mortality, using multivariable logistic regression. We identified 316 patients, of whom 46% died within 30-days. We developed a mortality score incorporating age, sex, platelet count, international normalised ratio, and observations on admission including the Glasgow Coma Scale, respiratory rate and blood pressure. The score was highly predictive of 30-day mortality with an area under the receiver operating curve of 0.7933 (95% CI 0.745-0.841). The optimal cut-point was a score ≥ 4, which had a sensitivity of 78.36% and a specificity of 67.59%. Patients with a score ≥ 4 had an odds ratio of 7.6 for 30-day mortality compared to those with a score < 4 (95% CI 4.56-12.49, p < 0.001). This simple, easy-to-use risk score calculator for patients admitted to hospital with COVID-19 is a strong predictor of 30-day mortality. Whilst requiring further external validation, it has the potential to guide prognosis for family and friends, and to identify patients at increased risk, who may require closer observation and more intensive early intervention.