A comparison of the validity of gas chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry analysis of urine samples II: amphetamine, methamphetamine, (±)-3,4-methylenedioxyamphetamine, (±)-3,4-methylenedioxymethamphetamine, (±)-3,4-methylenedioxyethylamphetamine, phencyclidine, and (±)-11-nor-9-carboxy-Δ⁹-tetrahydrocannabinol.
On November 25, 2008, the U.S. Department of Health and Human Services posted a final notice in the Federal Register authorizing the use of liquid chromatography-tandem mass spectrometry (LC-MS-MS) and other technologies in federally regulated workplace drug testing (WPDT) programs. To support this change, it is essential to explicitly demonstrate that LC-MS-MS, as a technology, can produce results at least as valid as gas chromatography (GC)-MS, the long-accepted standard in confirmatory analytical technologies for drugs of abuse. A series of manufactured control urine samples (n = 10 for each analyte) containing amphetamine, methamphetamine, (±)-3,4-methylenedioxyamphetamine, (±)-3,4-methylenedioxymethamphetamine, (±)-3,4-methylenedioxyethylamphetamine, phencyclidine, and (±)-11-nor-9-carboxy-Δ⁹-tetrahydrocannabinol at concentrations ranging from 10% to 2000% of federal cutoffs were analyzed with replication by five federally regulated laboratories using GC-MS and at RTI International using LC-MS-MS. Interference samples as described in the National Laboratory Certification Program 2009 Manual were analyzed by GC-MS and LC-MS-MS as well as previously confirmed urine specimens of WPDT origin. Matrix effects were assessed for LC-MS-MS. Results indicated that LC-MS-MS analysis produced results at least as precise, accurate, and specific as GC-MS for the analytes investigated in this study. Matrix effects, while evident, could be controlled by the use of matrix-matched controls and calibrators with deuterated internal standards.


Use of alternating logistic regression in studies of drug-use clustering.
This article describes the alternating logistic regression (ALR) method, and places this method in the context of other statistical approaches to the analysis of complex survey data, including the conditional form of logistic regression with matching on neighborhood characteristics. Unlike conditional logistic regression, the ALR method provides for an explicit estimation of the magnitude of clustering of drug use within neighborhoods and within subgroups of the neighborhood defined by male-female or age indicators, with and without covariate adjustments. The application of these ALR methods is illustrated with estimates for the magnitude of clustering of daily marijuana use and weekly marijuana use within neighborhoods of the United States, based on data from the National Household Survey on Drug Abuse samples from 1990 through 1996.


Malaria diagnostic testing and treatment practices in three different Plasmodium falciparum transmission settings in Tanzania: before and after a government policy change.
Patterns of decreasing malaria transmission intensity make presumptive treatment of malaria an unjustifiable approach in many African settings. The controlled use of anti-malarials after laboratory confirmed diagnosis is preferable in low endemic areas. Diagnosis may be facilitated by malaria rapid diagnostic tests (RDTs). In this study, the impact of a government policy change, comprising the provision of RDTs and advice to restrict anti-malarial treatment to RDT-positive individuals, was assessed by describing diagnostic behaviour and treatment decision-making in febrile outpatients <10 years of age in three hospitals in the Kagera and Mwanza Region in northern Tanzania. Prospective data from Biharamulo and Rubya Designated District Hospital (DDH) were collected before and after policy change, in Sumve DDH no new policy was implemented. Diagnosis of malaria was confirmed by RDT; transmission intensity was evaluated by a serological marker of malaria exposure in hospital attendees. Prior to policy change, there was no evident association between the actual level of transmission intensity and drug-prescribing behaviour. After policy change, there was a substantial decrease in anti-malarial prescription and an increase in prescription of antibiotics. The proportion of parasite-negative individuals who received anti-malarials decreased from 89.1% (244/274) to 38.7% (46/119) in Biharamulo and from 76.9% (190/247) to 10.0% (48/479) in Rubya after policy change. This study shows that an official policy change, where RDTs were provided and healthcare providers were advised to adhere to RDT results in prescribing drugs can be followed by more rational drug-prescribing behaviour. The current findings are promising for improving treatment policy in Tanzanian hospitals.


Signal amplification of microRNAs with modified strand displacement-based cycling probe technology.
Micro ribose nucleic acids (miRNAs) play an important role in biological processes such as cell differentiation, proliferation and apoptosis. Therefore, miRNAs are potentially a powerful marker for monitoring cancer and diagnosis. Here, we present sensitive signal amplification for miRNAs based on modified cycling probe technology with strand displacement amplification. miRNA was captured by the template coupled with beads, and then the first cycle based on SDA was repeatedly extended to the nicking end, which was produced by the extension reaction of miRNA. The products generated by SDA are captured by a molecular beacon (MB), which is designed to initiate the second amplification cycle, with a similar principle to the cycling probe technology (CPT), which is based on repeated digestion of the DNA-RNA hybrid by the RNase H. After one sample enrichment and two steps of signal amplification, 0.1 pM of let-7a can be detected. The miRNA assay exhibits a great dynamic range of over 100 orders of magnitude and high specificity to clearly discriminate a single base difference in miRNA sequences. This isothermal amplification does not require any special temperature control instrument. The assay is also about signal amplification rather than template amplification, therefore minimising contamination issues. In addition, there is no need for the reverse transcription (RT) process. Thus the amplification is suitable for miRNA detection.


A possible CO2 conducting and concentrating mechanism in plant stomata SLAC1 channel.
The plant SLAC1 is a slow anion channel in the membrane of stomatal guard cells, which controls the turgor pressure in the aperture-defining guard cells, thereby regulating the exchange of water vapour and photosynthetic gases in response to environmental signals such as drought, high levels of carbon dioxide, and bacterial invasion. Recent study demonstrated that bicarbonate is a small-molecule activator of SLAC1. Higher CO(2) and HCO(3)(-) concentration activates S-type anion channel currents in wild-type Arabidopsis guard cells. Based on the SLAC1 structure a theoretical model is derived to illustrate the activation of bicarbonate to SLAC1 channel. Meanwhile a possible CO(2) conducting and concentrating mechanism of the SLAC1 is proposed. The homology structure of Arabidopsis thaliana SLAC1 (AtSLAC1) provides the structural basis for study of the conducting and concentrating mechanism of carbon dioxide in SLAC1 channels. The pK(a) values of ionizable amino acid side chains in AtSLAC1 are calculated using software PROPKA3.0, and the concentration of CO(2) and anion HCO(3)(-) are computed based on the chemical equilibrium theory. The AtSLAC1 is modeled as a five-region channel with different pH values. The top and bottom layers of channel are the alkaline residue-dominated regions, and in the middle of channel there is the acidic region surrounding acidic residues His332. The CO(2) concentration is enhanced around 10(4) times by the pH difference between these regions, and CO(2) is stored in the hydrophobic region, which is a CO(2) pool. The pH driven CO(2) conduction from outside to inside balances the back electromotive force and maintain the influx of anions (e.g. Cl(-) and NO(3)(-)) from inside to outside. SLAC1 may be a pathway providing CO(2) for photosynthesis in the guard cells.


The career outcomes for doctors completing general practice vocational training 1990-1995.
While much has been published about the career outcomes of doctors who completed general practice vocational training prior to 1990, no evidence is currently available about those who have qualified since that time. To obtain information about the career paths of doctors who had completed general practice vocational training since 1990, and to compare the results with previously published data. Postal questionnaire survey of all doctors completing vocational training during the period 1990-1995 in three regions of the United Kingdom. The study examined current work status, career path since completion of training, desire for and experience of part-time training, degree of difficulty in choosing and following a career, and the degree to which certain factors impeded career choice. The overall response rate was 64.8%, although there was a significant difference between the response rates for men and women. While virtually all responders were employed, with the majority working in general practice, women were significantly less likely than men to be working as principals in general practice, for all cohorts. These results were very similar to those cohorts described in earlier studies. The career paths of doctors only became stable after about four years. Of those working in general practice, about 20% found it difficult to choose their career, and about 10% found it difficult to follow their career. Out-of-hours work was the major factor impeding career choice. Although they are taking longer to reach, the final career destinations of doctors completing vocational training since 1990 are no different from those of earlier cohorts.


'Well, It's the Risk of the Unknown… Right?': A Qualitative Study of Perceived Risks and Benefits of HIV Cure Research in the United States.
Biomedical research towards an HIV cure is advancing in the United States and elsewhere, yet little is known about perceptions of risks and benefits among potential study participants and other stakeholders. We conducted a qualitative study to explore perceived risks and benefits of investigational HIV cure research among people living with HIV (PLWHIV), biomedical HIV cure researchers, policy-makers and bioethicists. We conducted a qualitative research study using in-depth interviews with a purposive sample of PLWHIV, biomedical HIV cure researchers, policy-makers and bioethicists in 2015-2016. We analysed interview transcripts using thematic analysis anchored in grounded theory. We conducted and analyzed 36 key informant interviews. Qualitative analysis revealed four main findings. 1) Potential HIV cure study volunteers noted needing more information and education about the potential risks of HIV cure research. 2) Biomedical HIV cure researchers, policy-makers and bioethicists showed less awareness of social and financial risks of HIV cure research than PLWHIV. 3) Most respondents across the different categories of informants identified some risks that were too great to be acceptable in HIV cure research, although a subset of PLWHIV did not place an upper limit on acceptable risk. 4) PLWHIV showed a better awareness of potential psychological benefits of participating in HIV cure research than other groups of stakeholders. Our research suggests that PLWHIV have a variable understanding of the individual risks, sometimes substantial, associated with participating in biomedical HIV cure research studies. Community engagement and increased research literacy may help improve community understanding. Intensive informed consent procedures will be necessary for ethical study implementation. The current state of HIV cure research offers greater potential benefits to society than to participants. There is likely to be disagreement among regulators, researchers, clinicians, and potential participants about what constitutes acceptable risk for HIV cure studies.


Analysis of the effect of sentiment analysis on extracting adverse drug reactions from tweets and forum posts.
The abundance of text available in social media and health related forums along with the rich expression of public opinion have recently attracted the interest of the public health community to use these sources for pharmacovigilance. Based on the intuition that patients post about Adverse Drug Reactions (ADRs) expressing negative sentiments, we investigate the effect of sentiment analysis features in locating ADR mentions. We enrich the feature space of a state-of-the-art ADR identification method with sentiment analysis features. Using a corpus of posts from the DailyStrength forum and tweets annotated for ADR and indication mentions, we evaluate the extent to which sentiment analysis features help in locating ADR mentions and distinguishing them from indication mentions. Evaluation results show that sentiment analysis features marginally improve ADR identification in tweets and health related forum posts. Adding sentiment analysis features achieved a statistically significant F-measure increase from 72.14% to 73.22% in the Twitter part of an existing corpus using its original train/test split. Using stratified 10×10-fold cross-validation, statistically significant F-measure increases were shown in the DailyStrength part of the corpus, from 79.57% to 80.14%, and in the Twitter part of the corpus, from 66.91% to 69.16%. Moreover, sentiment analysis features are shown to reduce the number of ADRs being recognized as indications. This study shows that adding sentiment analysis features can marginally improve the performance of even a state-of-the-art ADR identification method. This improvement can be of use to pharmacovigilance practice, due to the rapidly increasing popularity of social media and health forums.


Real-world asthma management with inhaler devices in Switzerland-results of the asthma survey.
The aim of the Asthma Survey was to generate insights about the daily practice of physicians with regard to inhaler devices used for treating asthma under real-world conditions in Switzerland. A questionnaire was administered to 605 participating hospital- and practice-based Swiss physicians. Areas of interest were practical aspects of patient education, typical difficulties encountered when prescribing pressurized metered-dose inhalers (pMDI) and dry-powder inhalers (DPI), and reasons for physician preferences. Differences between the German-speaking part of Switzerland (D-CH) and French- and Italian-speaking parts of Switzerland (W-CH) linguistic regions were explored. Datasets from 529 physicians (291 D-CH and 238 W-CH) were suitable for analysis, 342 internists/general practitioners, 177 pulmonologists/allergologists, and 10 other. Approximately 90% of all participants declared being personally involved in providing inhaler device education to their patients. Practice assistants (33.0% vs. 9.2%, P<0.001) and pharmacists (6.9% vs. 19.7%, P<0.001) were more frequently involved in D-CH compared to W-CH. Patient skills with regard to inhalation technique were generally not monitored on a regular basis with only 34.0% of participants ensuring such checks at the scheduled visits. DPIs were overwhelmingly preferred over pMDI. Although the prevalence of typical handling errors was similar with both inhalers in the two regions, pMDIs were used more frequently in W-CH (P<0.001). Real-world asthma management and inhaler preferences differ between D-CH and W-CH. While the importance of patient education is widely acknowledged, inhalation skills monitoring remains suboptimal. The reasons for higher pMDI preference in W-CH compared to D-CH deserve further research.


Up-regulation of renal prostaglandin receptors in genetic salt-dependent hypertension.
Development of hypertension in Dahl salt-sensitive rats (DS) is accompanied by reduced renomedullary prostaglandin synthesis, which may be responsible for their lower natriuretic capacity. To examine the changes in renomedullary prostaglandin E2 synthesis, the effects of high (8.0%) and normal (0.6%) NaCl diets were examined in DS and in Dahl salt-resistant rats (DR). In response to an 8.0% NaCl diet, the number of prostaglandin E2 receptors in the renal outer medulla of DR increased (2.97 +/- 0.2 vs 2.18 +/- 0.2 pmol/mg on 0.6% NaCl diet) while no change was noted in their affinities (Kd, 9.5 +/- 0.2 vs 9.4 +/- 0.3 nM). Receptor number and affinity in the renal cortex, inner medulla, and liver of DR were not affected. In contrast, renomedullary receptors of DS had a lower affinity than those of age-matched DR (Kd, 13.9 +/- 0.2 nM on 0.6% NaCl diet and 14.0 +/- 0.3 nM on 8.0% NaCl diet) and did not increase in number after a high salt diet. This apparent inability of DS to modulate prostaglandin receptors may contribute to their susceptibility to salt-induced hypertension.


Gene selection for optimal prediction of cell position in tissues from single-cell transcriptomics data.
Single-cell RNA-sequencing (scRNAseq) technologies are rapidly evolving. Although very informative, in standard scRNAseq experiments, the spatial organization of the cells in the tissue of origin is lost. Conversely, spatial RNA-seq technologies designed to maintain cell localization have limited throughput and gene coverage. Mapping scRNAseq to genes with spatial information increases coverage while providing spatial location. However, methods to perform such mapping have not yet been benchmarked. To fill this gap, we organized the DREAM Single-Cell Transcriptomics challenge focused on the spatial reconstruction of cells from the Drosophila embryo from scRNAseq data, leveraging as silver standard, genes with in situ hybridization data from the Berkeley Drosophila Transcription Network Project reference atlas. The 34 participating teams used diverse algorithms for gene selection and location prediction, while being able to correctly localize clusters of cells. Selection of predictor genes was essential for this task. Predictor genes showed a relatively high expression entropy, high spatial clustering and included prominent developmental genes such as gap and pair-rule genes and tissue markers. Application of the top 10 methods to a zebra fish embryo dataset yielded similar performance and statistical properties of the selected genes than in the Drosophila data. This suggests that methods developed in this challenge are able to extract generalizable properties of genes that are useful to accurately reconstruct the spatial arrangement of cells in tissues.


Functional interaction between Lypd6 and nicotinic acetylcholine receptors.
Nicotinic acetylcholine receptors (nAChRs) affect multiple physiological functions in the brain and their functions are modulated by regulatory proteins of the Lynx family. Here, we report for the first time a direct interaction of the Lynx protein LY6/PLAUR domain-containing 6 (Lypd6) with nAChRs in human brain extracts, identifying Lypd6 as a novel regulator of nAChR function. Using protein cross-linking and affinity purification from human temporal cortical extracts, we demonstrate that Lypd6 is a synaptically enriched membrane-bound protein that binds to multiple nAChR subtypes in the human brain. Additionally, soluble recombinant Lypd6 protein attenuates nicotine-induced hippocampal inward currents in rat brain slices and decreases nicotine-induced extracellular signal-regulated kinase phosphorylation in PC12 cells, suggesting that binding of Lypd6 is sufficient to inhibit nAChR-mediated intracellular signaling. We further show that perinatal nicotine exposure in rats (4 mg/kg/day through minipumps to dams from embryonic day 7 to post-natal day 21) significantly increases Lypd6 protein levels in the hippocampus in adulthood, which did not occur after exposure to nicotine in adulthood only. Our findings suggest that Lypd6 is a versatile inhibitor of cholinergic signaling in the brain, and that Lypd6 is dysregulated by nicotine exposure during early development. Regulatory proteins of the Lynx family modulate the function of nicotinic receptors (nAChRs). We report for the first time that the Lynx protein Lypd6 binds to nAChRs in human brain extracts, and that recombinant Lypd6 decreases nicotine-induced ERK phosphorylation and attenuates nicotine-induced hippocampal inward currents. Our findings suggest that Lypd6 is a versatile inhibitor of cholinergic signaling in the brain.


The assessment of social adjustment. An update.
Since a review of 15 social adjustment scales appeared six years ago in the ARCHIVES, a number of new scales have been developed and tested. Twelve new scales, three written self-administered and nine interviewer-administered scales, are considered to be useful in clinical and epidemiologic research in psychiatry. Future research in this area should include development of scales for use with children, adolescents, and the elderly; revision of scales to reflect changes in traditional roles, especially among women; clarification of the concepts underlying social functioning; and systematic explication and measurement of these concepts.


[Spectrophotometric determination of oleanolic acid and saponins from quinoa (Chenopodium quinoa Willd, Kancolla variety)].
Saponins were extracted from quinoa (Chenopodium quinoa Willd, Kancolla variety) by refluxing with a methanol-water (4:1) mixture. Once the methanol was evaporated, the remaining residue was treated following Honerlagen and Tretter's method with only slight modifications. The extract was then hydrolyzed with 12N sulfuric acid in a 1:1 dioxane-water system at 110 degrees C for 1.5 hr. The sapogenins were extracted with chloroform, concentrated and some microliters (equivalent to 121 mg of quinoa) were spotted, against an oleanolic acid standard, on a silicagel g plate and developed with a chloroform-acetone-benzene (80:20:10; v/v) mixture. The spots were located by iodine vapor, and the band whose Rf was similar to that of the oleanolic acid, was scraped into a glass column, eluted with chloroform, dried out, dissolved in 1 ml of glacial acetic acid, treated with 4 ml of (1:1; v/v) sulfuric acid:glacial acetic acid mixture, heated in a water bath at 60 degrees C for 25 minutes, cooled and taken to the spectrophotometer where it was read at a wave length of 527 nm against a reagent blank. Under the same conditions, the oleanolic acid employed as a standard showed a linearity in the range of 60 to 480 micrograms. The oleanolic acid percentage has been determined (0.269 +/- 0.025) in quinoa, and the content of saponins estimated using a conversion factor found by gas chromatography and expressed in the following relationship: % Saponin = (8.5204) x (% oleanolic acid) The sapogenin extract obtained - analyzed by this method - showed an error of 10.7% in relation to its gas chromatography determination.


Use of seroconversion panels to estimate delay in detection of anti-human immunodeficiency virus antibodies by enzyme-linked immunosorbent assay of pooled compared to singleton serum samples.
The transfusion of unsafe blood worldwide accounts for 5 to 15% of new human immunodeficiency virus (HIV) infections, most of which occur in sub-Saharan Africa. While developed countries now apply PCR testing of pooled samples, some developing countries still do not have universal screening policies. More efficient low-cost procedures for the screening of pooled samples have the potential to encourage mass screening efforts in resource-poor settings. The aim of this study was to estimate the delay in the detection of HIV antibodies in pooled serum samples compared to that in singleton serum samples by enzyme-linked immunosorbent assay (ELISA) and to evaluate the risk of transfusion-transmitted HIV infection during the window period. Serial blood samples obtained from five HIV seroconversion panels were mixed with HIV-seronegative blood samples to create pools of 6, 12, 16, 24, 32, and 48 samples. The delay in detection of the first anti-HIV antibody-positive sample in tests with pooled samples was calculated for each pool size and compared to that obtained by testing of singleton samples and statistically evaluated by a robust log-linear regression analysis. The risk of a false-negative (FN) result caused by dilution was estimated by use of the incidence risk/window period model. The additional risk of transmission related to ELISA screening of pooled samples for HIV did not exceed 9% of the current risk of an FN result (estimated to be 1/1,067,000). The countries with virus prevalence rates in donors of less than 15% are expected to save up to 30% in the number of tests. ELISA screening of pooled samples could be considered in settings where the testing of blood supplies for HIV is not routinely done.


Publication guidelines for quality improvement studies in health care: evolution of the SQUIRE project.
In 2005 we published draft guidelines for reporting studies of quality improvement, as the initial step in a consensus process for development of a more definitive version. The current article contains the revised version, which we refer to as standards for quality improvement reporting excellence (SQUIRE). This narrative progress report summarises the special features of improvement that are reflected in SQUIRE, and describes major differences between SQUIRE and the initial draft guidelines. It also briefly describes the guideline development process; considers the limitations of and unresolved questions about SQUIRE; describes ancillary supporting documents and alternative versions under development; and discusses plans for dissemination, testing, and further development of SQUIRE.


Fine needle aspiration cytology in childhood TB lymphadenitis.
To evaluate the efficacy of Fine Needle Aspiration Cytology (FNAC) to diagnose Tuberculous (TB) lymphadenitis with compare to excision biopsy and to correlate TB lymphadenitis with clinical, cytological, radiological and mantoux test features. This was a prospective correlational study. FNAC was done by a pediatrician for 135 children with persisting lymphadenitis after two weeks of antibiotic therapy in the period of January 2005 to June 2006 and compared with excision biopsy in a tertiary care hospital. Forty Six cases (34.07%) were TB lymphadenitis diagnosed by FNAC. Excision biopsy and cytological correlation was done in 100 cases. Sensitivity, specificity and diagnostic accuracy for TB lymphadenitis were found to be 98%, 100% and 99% respectively. Positive and negative predictive values were 100 and 98 respectively. Large (>2cm) (86.9%), multiple (52.1%), matted (47.8%), posterior cervical and submandibular group nodes with history of contact (P=0.0016), positive mantoux test (P=0.0001) and Grade III and IV Protein Energy Malnutrition (PEM) (P=0.0041) were significantly seen in TB lymphadenitis. Ziehl Neelson staining for Acid Fast Bacilli (AFB) was positive in 32.5% cases of TB Lymphadenitis. Pediatrician himself can do FNAC which is an excellent first line method to diagnose TB lymphadenitis and it has equal accuracy to excision biopsy.


The puzzle of self-reported weight gain in a month of fasting (Ramadan) among a cohort of Saudi families in Jeddah, Western Saudi Arabia.
During Ramadan fast, approximately one billion Muslims abstain from food and fluid between the hours of sunrise to sunset, and usually eat a large meal after sunset and another meal before sunrise. Many studies reported good health-related outcomes of fasting including weight loss. The objective of this study is to identify the local pattern of expenditure on food consumption, dietary habits during Ramadan and correlate that to self-reported weight gain after Ramadan in a group of families in Jeddah, Western Saudi Arabia. A Cross-section study using a pre-designed questionnaire to identify the local pattern of expenditure on food consumption, dietary habits during Ramadan and correlate that to self-reported weight gain after Ramadan in a representative cohort of Saudis living in Jeddah. It was piloted on 173 nutrition students and administered by them to their families. A total of 173 Saudi families were interviewed. One out of 5 indicated that their expenditure increases during Ramadan. Approximately two thirds of the respondents (59.5%) reported weight gain after Ramadan. When asked about their perspective explanations for that: 40% attributed that to types of foods being rich in fat and carbohydrates particularly date in (Sunset meal) 97.7% and rice in (Dawn meal) 80.9%. One third (31.2%) indicated that it was due to relative lack of physical exercise in Ramadan and 14.5% referred that to increase in food consumption. Two thirds (65.2%) of those with increased expenditure reported weight gain. Surprisingly weight gain and not weight loss was reported after Ramadan by Saudis which indicates timely needed life-style and dietary modification programs for a population which reports one of the highest prevalence rates of diabetes.


Effect of hydroxychloroquine and characterization of autophagy in a mouse model of endometriosis.
In endometriosis, the increased survival potential of shed endometrial cells (which normally undergo anoikis) is suggested to promote lesion development. One mechanism that may alter anoikis is autophagy. Using an autophagic flux inhibitor hydroxychloroquine (HCQ), we identified that it reduces the in vitro survival capacity of human endometriotic and endometrial T-HESC cells. We also identified that HCQ could decrease lesion numbers and disrupt lesion histopathology, as well as increase the levels of peritoneal macrophages and the IP-10 (10 kDa interferon-γ-induced protein) chemokine in a mouse model of endometriosis. We noted that RNA levels of a subset of autophagic markers were reduced in lesions relative to uterine horns from endometriosis-induced (untreated) mice. In addition, the RNA levels of autophagic markers were decreased in uterine horns of endometriosis-induced mice compared with those from controls. However, we noted that protein expression of LC3B (microtubule-associated protein 1 light-chain 3β; an autophagic marker) was increased in uterine horns of endometriosis-induced mice compared with uterine horns of controls. By immunohistochemical staining of a human endometriosis-focused tissue microarray, we observed LC3B expression predominantly in epithelial relative to stromal cells in both eutopic and ectopic endometria. Via transmission electron microscopy, cells from eutopic endometria of endometriosis-induced mice contained more lipid droplets (rather than autophagosomes) compared with uterine horns from controls. Collectively, our findings indicate that the autophagic pathway is dysregulated in both ectopic and eutopic endometrium in a murine model of endometriosis and that HCQ has potential as a therapeutic agent for women afflicted with endometriosis.


Melatonin inhibits both ER alpha activation and breast cancer cell proliferation induced by a metalloestrogen, cadmium.
Cadmium (Cd) is a heavy metal affecting human health both through environmental and occupational exposure. There is evidence that Cd accumulates in several organs and is carcinogenic to humans. In vivo, Cd mimics the effect of estrogens in the uterus and mammary gland. In estrogen-responsive breast cancer cell lines, Cd stimulates proliferation and can also activate the estrogen receptor independent of estradiol. The ability of this metalloestrogen to increase gene expression in MCF7 cells is blocked by anti-estrogens suggesting that the activity of these compounds is mediated by ER alpha. The aims of this work were to test whether melatonin inhibits Cd-induced proliferation in MCF7 cells, and also to study whether melatonin specifically inhibits Cd-induced ER alpha transactivation. We show that melatonin prevents the Cd-induced growth of synchronized MCF7 breast cancer cells. In transient transfection experiments, we prove that both ER alpha- and ER beta-mediated transcription are stimulated by Cd. Melatonin is a specific inhibitor of Cd-induced ER alpha-mediated transcription in both estrogen response elements (ERE)- and AP1-containing promoters, whereas ER beta-mediated transcription is not inhibited by the pineal indole. Moreover, the mutant ER alpha-(K302G, K303G), unable to bind calmodulin, is activated by Cd but becomes insensitive to melatonin treatment. These results proved that melatonin inhibits MCF7 cell growth induced by Cd and abolishes the stimulatory effect of the heavy metal in cells expressing ER alpha at both ERE-luc and AP1-luc sites. We can infer from these experiments that melatonin regulates Cd-induced transcription in both ERE- and AP1 pathways. These results also reinforce the hypothesis of the anti-estrogenic properties of melatonin as a valuable tool in breast cancer therapies.


Effect of particle morphology on emitted dose of fatty acid-treated disodium cromoglycate powder aerosols.
These studies assess the quantity and morphology of the emitted aerosolized dose of irregularly shaped disodium cromoglycate particles in the fine particle fraction using in vitro methods. Disodium cromoglycate was treated with a homologous series of saturated fatty acids, between C8 and C18, in a range of concentrations. The products of these treatments were powders with a variety of particle size, shape, and aggregation characteristics. Samples of these powders were loaded in gelatin capsules, generated as aerosols from a Rotahaler and collected in a two-stage liquid impinger or eight-stage inertial impactor. Particles were examined directly by scanning electron microscopy and subsequently the images were analyzed to define morphology. The aerodynamic fine-particle fraction determined by the two-stage impinger increased approximately twofold with lauric acid treatment (0.0317 g/g, 6.7%) and threefold with stearic acid treatment (0.58 g/g; 9.7%) compared with disodium cromoglycate alone (0 g/g, 3.56%). The lauric acid formulation appeared to alter deposition primarily by changing particle morphology. Stearic acid altered particle shape to some extent and the increase in the fine-particle fraction appeared to be attributable to improved particle dispersion properties. The uncontrolled presence of irregular-shaped particles can introduce dosing errors due to effects on dispersion and aerodynamic behavior. Conversely, controlled particle morphology and size may be employed to optimize the dose delivered to the lungs particularly if particle-particle and particle-surface interactions can be minimized.


Bacteriophages are the major drivers of Shigella flexneri serotype 1c genome plasticity: a complete genome analysis.
Shigella flexneri is the primary cause of bacillary dysentery in the developing countries. S. flexneri serotype 1c is a novel serotype, which is found to be endemic in many developing countries, but little is known about its genomic architecture and virulence signatures. We have sequenced for the first time, the complete genome of S. flexneri serotype 1c strain Y394, to provide insights into its diversity and evolution. We generated a high-quality reference genome of S. flexneri serotype 1c using the hybrid methods of long-read single-molecule real-time (SMRT) sequencing technology and short-read MiSeq (Illumina) sequencing technology. The Y394 chromosome is 4.58 Mb in size and shares the basic genomic features with other S. flexneri complete genomes. However, it possesses unique and highly modified O-antigen structure comprising of three distinct O-antigen modifying gene clusters that potentially came from three different bacteriophages. It also possesses a large number of hypothetical unique genes compared to other S. flexneri genomes. Despite a high level of structural and functional similarities of Y394 genome with other S. flexneri genomes, there are marked differences in the pathogenic islands. The diversity in the pathogenic islands suggests that these bacterial pathogens are well adapted to respond to the selection pressures during their evolution, which might contribute to the differences in their virulence potential.


A GRF-GIF chimeric protein improves the regeneration efficiency of transgenic plants.
The potential of genome editing to improve the agronomic performance of crops is often limited by low plant regeneration efficiencies and few transformable genotypes. Here, we show that expression of a fusion protein combining wheat GROWTH-REGULATING FACTOR 4 (GRF4) and its cofactor GRF-INTERACTING FACTOR 1 (GIF1) substantially increases the efficiency and speed of regeneration in wheat, triticale and rice and increases the number of transformable wheat genotypes. GRF4-GIF1 transgenic plants were fertile and without obvious developmental defects. Moreover, GRF4-GIF1 induced efficient wheat regeneration in the absence of exogenous cytokinins, which facilitates selection of transgenic plants without selectable markers. We also combined GRF4-GIF1 with CRISPR-Cas9 genome editing and generated 30 edited wheat plants with disruptions in the gene Q (AP2L-A5). Finally, we show that a dicot GRF-GIF chimera improves regeneration efficiency in citrus, suggesting that this strategy can be applied to dicot crops.


Acromegaly: an endocrine society clinical practice guideline.
The aim was to formulate clinical practice guidelines for acromegaly. The Task Force included a chair selected by the Endocrine Society Clinical Guidelines Subcommittee (CGS), five experts in the field, and a methodologist. The authors received no corporate funding or remuneration. This guideline is cosponsored by the European Society of Endocrinology. This evidence-based guideline was developed using the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) system to describe both the strength of recommendations and the quality of evidence. The Task Force reviewed primary evidence and commissioned two additional systematic reviews. One group meeting, several conference calls, and e-mail communications enabled consensus. Committees and members of the Endocrine Society and the European Society of Endocrinology reviewed drafts of the guidelines. Using an evidence-based approach, this acromegaly guideline addresses important clinical issues regarding the evaluation and management of acromegaly, including the appropriate biochemical assessment, a therapeutic algorithm, including use of medical monotherapy or combination therapy, and management during pregnancy.


RFDCR: Automated brain lesion segmentation using cascaded random forests with dense conditional random fields.
Segmentation of brain lesions from magnetic resonance images (MRI) is an important step for disease diagnosis, surgical planning, radiotherapy and chemotherapy. However, due to noise, motion, and partial volume effects, automated segmentation of lesions from MRI is still a challenging task. In this paper, we propose a two-stage supervised learning framework for automatic brain lesion segmentation. Specifically, in the first stage, intensity-based statistical features, template-based asymmetric features, and GMM-based tissue probability maps are used to train the initial random forest classifier. Next, the dense conditional random field optimizes the probability maps from the initial random forest classifier and derives the whole tumor regions referred as the region of interest (ROI). In the second stage, the optimized probability maps are further intergraded with features from the intensity-based statistical features and template-based asymmetric features to train subsequent random forest, focusing on classifying voxels within the ROI. The output probability maps will be also optimized by the dense conditional random fields, and further used to iteratively train a cascade of random forests. Through hierarchical learning of the cascaded random forests and dense conditional random fields, the multimodal local and global appearance information is integrated with the contextual information, and the output probability maps are improved layer by layer to finally obtain optimal segmentation results. We evaluated the proposed method on the publicly available brain tumor datasets BRATS 2015 & BRATS 2018, as well as the ischemic stroke dataset ISLES 2015. The results have shown that our framework achieves competitive performance compared to the state-of-the-art brain lesion segmentation methods. In addition, contralateral difference and skewness were identified as the important features in the brain tumor and ischemic stroke segmentation tasks, which conforms to the knowledge and experience of medical experts, further reflecting the reliability and interpretability of our framework.


Humic substances of varying types increase survivorship of the freshwater shrimp Caridina sp. D to acid mine drainage.
Differences relating to the ability of various types of humic substances (HS) to influence toxicity of pollutants have been reported in the literature, but there still remains a gap in understanding whether various HS will have the same influence on the toxicity of acid mine drainage (AMD). This study investigated differences in the ability of Aldrich humic acid (AHA), Suwannee River humic acid and Suwannee River fulvic acid to decrease toxicity of AMD to the freshwater shrimp (Caridina sp. D). Toxicity tests were conducted over 96 h and used Mount Morgan open pit water as source of AMD and Dee River water as control/diluents. Concentrations of 0-4 % AMD at 0 mg/L HS, 10 mg/L AHA, 10 mg/L Suwannee River humic acid and 10 mg/L Suwannee River fulvic acid were used. Significantly higher survival of shrimp was recorded in the HS treatments compared with the treatment containing no HS. No significant differences were found among HS type. HS considerably increased LC50 values irrespective of type, from 1.29 (0 mg/L HS) to 2.12 % (AHA); 2.19 (Suwannee River humic acid) and 2.22 % (Suwannee River fulvic acid). These results support previous work that HS decrease the toxicity of AMD to freshwater organisms, but with the novel finding that this ability occurs irrespective of HS type. These results increase the stock of knowledge regarding HS and may contribute to a possible remediation option for AMD environments.


Racial differences in erythrocyte cation transport.
Erythrocyte contents and ouabain-insensitive transport pathways were measured in 120 white and black normotensives and hypertensives. Mean maximal sodium-stimulated lithium-sodium countertransport rate was higher in white hypertensives than in white normotensives, and countertransport was significantly positively correlated with mean arterial pressure in whites. Values similar to those in white normotensives were found in both black normotensives and hypertensives, and countertransport was not significantly correlated with blood pressure in blacks. The rate constant for passive lithium efflux was greater in whites as compared to blacks, and the difference was not related to blood pressure level or sex. Ouabain-insensitive, furosemide-sensitive sodium and potassium effluxes were not found to be altered in hypertension. Furosemide-sensitive sodium efflux rate was lower in blacks but furosemide-sensitive potassium efflux was not similarly depressed. While white subjects demonstrated a close correlation between sodium and potassium effluxes, blacks did not. Further study of these differences in the cellular metabolism of sodium and potassium may provide clues to the pathogenesis of racial dissimilarities in total body sodium handling.


From in silico to in vitro: modelling and production of Trichoderma reesei endoglucanase 1 and its mutant in Pichia pastoris.
In this study, a major cellulase, namely endoglucanase 1 (EGI) from Trichoderma reesei was mutated by the introduction of four different lysine and glycine rich loops to create a hotspot for directed crosslinking of EGI away from the active site. The impact of the inserted loops on the stability of the enzyme was analyzed using molecular dynamics (MD) and the effect on the active site was studied using molecular mechanics (MM) simulations. The best loop mutation predicted in silico (EGI_L5) was introduced to EGI via site directed mutagenesis. The loop mutant EGI_L5 and EGI were both expressed in Pichia pastoris. Enzymes were characterized and their activities against soluble substrates such as CMC and 4-MUC were determined. Both enzymes exhibited similar pH and temperature activity and thermal stability profiles. Moreover, specific activity of EGI_L5 against 4-MUC was found to be the same as the native enzyme.


Differences in affinity of binding of lymphocytic choriomeningitis virus strains to the cellular receptor alpha-dystroglycan correlate with viral tropism and disease kinetics.
alpha-Dystroglycan (alpha-DG) was recently identified as a receptor for lymphocytic choriomeningitis virus (LCMV) and several other arenaviruses, including Lassa fever virus (W. Cao, M. D. Henry, P. Borrow, H. Yamada, J. H. Elder, E. V. Ravkov, S. T. Nichol, R. W. Compans, K. P. Campbell, and M. B. A. Oldstone, Science 282:2079-2081, 1998). Data presented in this paper indicate that the affinity of binding of LCMV to alpha-DG determines viral tropism and the outcome of infection in mice. To characterize this relationship, we evaluated the interaction between alpha-DG and several LCMV strains, variants, and reassortants. These viruses could be divided into two groups with respect to affinity of binding to alpha-DG, dependence on this protein for cell entry, viral tropism, and disease course. Viruses that exhibited high-affinity binding to alpha-DG displayed a marked dependence on alpha-DG for cell entry and were blocked from infecting mouse 3T6 fibroblasts by 1 to 4 nM soluble alpha-DG. In addition, high-affinity binding to alpha-DG correlated with an ability to infiltrate the white pulp (T-dependent) area of the spleen, cause ablation of the cytotoxic T-lymphocyte (CTL) response by day 7 postinfection, and establish a persistent infection. In contrast, viruses with a lower affinity of binding to alpha-DG were only partially inhibited from infecting alpha-DG(-/-) embryonic stem cells and required a concentration of soluble alpha-DG higher than 100 nM to prevent infection of mouse 3T6 fibroblasts. These viruses that bound at low affinity were mainly restricted to the splenic red pulp, and the host generated an effective CTL response that rapidly cleared the infection. Reassortants of viruses that bound to alpha-DG at high and low affinities were used to map genes responsible for the differences described to the S RNA, containing the virus attachment protein glycoprotein 1.


High-precision rotation angle measurement method based on monocular vision.
To accurately measure the attitude angles (pitch, roll, and yaw) of a rigid object that rotates in a space, we propose a high-precision rotation angle measurement method based on monocular vision. This method combines camera self-calibration, multiview geometry, and 3D measurement. This monocular vision measuring system consists of an area scan CCD, a prime lens, and a spots array target, which are fixed on the measured object. We can calculate the rotation angle according to the rebuilt rotating spots array target by using this monocular vision measuring system. The measurement precision of rotation angle can reach 1 arc sec in this paper's experiments. This method has high measurement precision and good stability. Therefore we can widely use this method in machinery manufacturing, engineering measurement, aerospace, and the military.


Geographic disparities in late-stage cancer diagnosis: multilevel factors and spatial interactions.
In 2009 in the United States, breast cancer was the most common cancer in women, and colorectal cancer was the third most common cancer in both men and women. Currently, over 40% of these cancers are diagnosed at an advanced stage, which results in higher morbidity and mortality than would obtain with optimal cancer screening utilization. To provide information that might improve these cancer outcomes we use spatial analysis to answer questions related to both Why and Where disparities in late-stage cancer diagnoses are observed. In examining Why, we include state level characteristics reflecting characteristics of states' cancer control planning, insurance markets and managed care environments to help model the spatial heterogeneity from place to place. To answer questions related to Where disparities are observed, we generate county level predictions of late-stage cancer rates from a random-intercept multilevel model estimated on the population data from 11 pooled SEER Registries. The findings allow for comparisons across states that reveal logical starting points for a national effort to control cancer.


Initiating an undiagnosed diseases program in the Western Australian public health system.
New approaches are required to address the needs of complex undiagnosed diseases patients. These approaches include clinical genomic diagnostic pipelines, utilizing intra- and multi-disciplinary platforms, as well as specialty-specific genomic clinics. Both are advancing diagnostic rates. However, complementary cross-disciplinary approaches are also critical to address those patients with multisystem disorders who traverse the bounds of multiple specialties and remain undiagnosed despite existing intra-specialty and genomic-focused approaches. The diagnostic possibilities of undiagnosed diseases include genetic and non-genetic conditions. The focus on genetic diseases addresses some of these disorders, however a cross-disciplinary approach is needed that also simultaneously addresses other disorder types. Herein, we describe the initiation and summary outcomes of a public health system approach for complex undiagnosed patients - the Undiagnosed Diseases Program-Western Australia (UDP-WA). Briefly the UDP-WA is: i) one of a complementary suite of approaches that is being delivered within health service, and with community engagement, to address the needs of those with severe undiagnosed diseases; ii) delivered within a public health system to support equitable access to health care, including for those from remote and regional areas; iii) providing diagnoses and improved patient care; iv) delivering a platform for in-service and real time genomic and phenomic education for clinicians that traverses a diverse range of specialties; v) retaining and recapturing clinical expertise; vi) supporting the education of junior and more senior medical staff; vii) designed to integrate with clinical translational research; and viii) is supporting greater connectedness for patients, families and medical staff. The UDP-WA has been initiated in the public health system to complement existing clinical genomic approaches; it has been targeted to those with a specific diagnostic need, and initiated by redirecting existing clinical and financial resources. The UDP-WA supports the provision of equitable and sustainable diagnostics and simultaneously supports capacity building in clinical care and translational research, for those with undiagnosed, typically rare, conditions.


Estimating the entropy rate of spike trains via Lempel-Ziv complexity.
Normalized Lempel-Ziv complexity, which measures the generation rate of new patterns along a digital sequence, is closely related to such important source properties as entropy and compression ratio, but, in contrast to these, it is a property of individual sequences. In this article, we propose to exploit this concept to estimate (or, at least, to bound from below) the entropy of neural discharges (spike trains). The main advantages of this method include fast convergence of the estimator (as supported by numerical simulation) and the fact that there is no need to know the probability law of the process generating the signal. Furthermore, we present numerical and experimental comparisons of the new method against the standard method based on word frequencies, providing evidence that this new approach is an alternative entropy estimator for binned spike trains.


Csk-binding protein can regulate Lyn signals controlling cell morphology.
The Src family kinase Lyn is involved in differentiation signals emanating from activated erythropoietin (Epo) receptors, it interacts with COOH-terminal Src kinase-binding protein (Cbp), an adaptor protein that recruits negative regulators COOH-terminal Src kinase (Csk) and suppressor of cytokine signaling-1 (SOCS1). Lyn phosphorylates Cbp on several tyrosine residues, including Tyr314, which recruits Csk/SOCS1, as well as Tyr381 and Tyr409 that bind Lyns own SH2 domain. We show that Cbp alters not only the ability of erythroid cells to differentiate but also their colony morphology. Consequently, we detailed the ability of Cbp to interact with and influence Lyns ability to initiate changes in cellular architecture, which affect cell-cell and cell-substratum interactions. Over-expression of active Lyn promotes filopodia formation while inactive Lyn promotes lamellipodia formation. Conversely, Cbp over-expression, which inhibits Lyn activity, promotes lamellipodia formation, while Cbp mutants preventing its interaction/signaling consequently allow Lyn to promote filopodia formation. Thus, the Lyn-Cbp pathway and subsequent regulation of Lyn signaling and cell morphology involves a dynamic and complex series of interactions.


Strictly positive-definite spike train kernels for point-process divergences.
Exploratory tools that are sensitive to arbitrary statistical variations in spike train observations open up the possibility of novel neuroscientific discoveries. Developing such tools, however, is difficult due to the lack of Euclidean structure of the spike train space, and an experimenter usually prefers simpler tools that capture only limited statistical features of the spike train, such as mean spike count or mean firing rate. We explore strictly positive-definite kernels on the space of spike trains to offer both a structural representation of this space and a platform for developing statistical measures that explore features beyond count or rate. We apply these kernels to construct measures of divergence between two point processes and use them for hypothesis testing, that is, to observe if two sets of spike trains originate from the same underlying probability law. Although there exist positive-definite spike train kernels in the literature, we establish that these kernels are not strictly definite and thus do not induce measures of divergence. We discuss the properties of both of these existing nonstrict kernels and the novel strict kernels in terms of their computational complexity, choice of free parameters, and performance on both synthetic and real data through kernel principal component analysis and hypothesis testing.


Conjugated linoleic acid protects against gliadin-induced depletion of intestinal defenses.
The involvement of oxidative stress in gluten-induced toxicity has been evidenced in vitro and in clinical studies but has never been examined in vivo. We recently demonstrated the protective activity of conjugated linoleic acid (CLA), which functions by the activation of nuclear factor erythroid 2-related factor2 (Nrf2), a key transcription factor for the synthesis of antioxidant and detoxifying enzymes (phase 2). Here, we evaluate the involvement of nuclear factor erythroid 2-related factor2 in gliadin-mediated toxicity in human Caco-2 intestinal cells and in gliadin-sensitive human leukocyte antigen-DQ8 transgenic mice (DQ8) and the protective activity of CLA. Gliadin effects in differentiated Caco-2 cells and in DQ8 mice, fed with a gliadin-containing diet with or without CLA supplementation, were evaluated by combining enzymatic, immunochemical, immunohistochemical, and quantitative real-time PCR (qRT-PCR) assays. Gliadin toxicity was accompanied by downregulation of phase 2 and elevates proteasome-acylpeptide hydrolase activities in vitro and in vivo. Notably, gliadin was unable to generate severe oxidative stress extent or pathological consequences in DQ8 mice intestine comparable to those found in celiac patients and the alterations produced were hampered by CLA. The beneficial effects of CLA against the depletion of crucial intestinal cytoprotective defenses indicates a novel nutritional approach for the treatment of intestinal disease associated with altered redox homeostasis.


Serum brain-derived neurotrophic factor and the risk for dementia: the Framingham Heart Study.
In animal studies, brain-derived neurotrophic factor (BDNF) has been shown to impact neuronal survival and function and improve synaptic plasticity and long-term memory. Circulating BDNF levels increase with physical activity and caloric restriction, thus BDNF may mediate some of the observed associations between lifestyle and the risk for dementia. Some prior studies showed lower circulating BDNF in persons with Alzheimer disease (AD) compared with control participants; however, it remains uncertain whether reduced levels precede dementia onset. To examine whether higher serum BDNF levels in cognitively healthy adults protect against the future risk for dementia and AD and to identify potential modifiers of this association. Framingham Study original and offspring participants were followed up from 1992 and 1998, respectively, for up to 10 years. We used Cox models to relate BDNF levels to the risk for dementia and AD and adjusted for potential confounders. We also ran sensitivity analyses stratified by sex, age, and education, as well as related BDNF genetic variants to AD risk. This community-based, prospective cohort study involved 2131 dementia-free participants aged 60 years and older (mean [SD] age, 72 [7] years; 56% women). Ten-year incidence of dementia and AD. During follow-up, 140 participants developed dementia, 117 of whom had AD. Controlling for age and sex, each standard-deviation increment in BDNF was associated with a 33% lower risk for dementia and AD (P = .006 and P = .01, respectively) and these associations persisted after additional adjustments. Compared with the bottom quintile, BDNF levels in the top quintile were associated with less than half the risk for dementia and AD (hazard ratio, 0.49; 95% CI, 0.28-0.85; P = .01; and hazard ratio, 0.46; 95% CI, 0.24-0.86; P = .02, respectively). These associations were apparent only among women, persons aged 80 years and older, and those with college degrees (hazard ratios for AD: 0.65, [95% CI, 0.50-0.85], P = .001; 0.63 [95% CI, 0.47-0.85], P = .002; and 0.27 [95% CI, 0.11-0.65], P = .003, respectively). Brain-derived neurotrophic factor genetic variants were not associated with AD risk. Higher serum BDNF levels may protect against future occurrence of dementia and AD. Our findings suggest a role for BDNF in the biology and possibly in the prevention of dementia and AD, especially in select subgroups of women and older and more highly educated persons.


Diurnal Transcriptome and Gene Network Represented through Sparse Modeling in Brachypodium distachyon.
We report the comprehensive identification of periodic genes and their network inference, based on a gene co-expression analysis and an Auto-Regressive eXogenous (ARX) model with a group smoothly clipped absolute deviation (SCAD) method using a time-series transcriptome dataset in a model grass, Brachypodium distachyon. To reveal the diurnal changes in the transcriptome in B. distachyon, we performed RNA-seq analysis of its leaves sampled through a diurnal cycle of over 48 h at 4 h intervals using three biological replications, and identified 3,621 periodic genes through our wavelet analysis. The expression data are feasible to infer network sparsity based on ARX models. We found that genes involved in biological processes such as transcriptional regulation, protein degradation, and post-transcriptional modification and photosynthesis are significantly enriched in the periodic genes, suggesting that these processes might be regulated by circadian rhythm in B. distachyon. On the basis of the time-series expression patterns of the periodic genes, we constructed a chronological gene co-expression network and identified putative transcription factors encoding genes that might be involved in the time-specific regulatory transcriptional network. Moreover, we inferred a transcriptional network composed of the periodic genes in B. distachyon, aiming to identify genes associated with other genes through variable selection by grouping time points for each gene. Based on the ARX model with the group SCAD regularization using our time-series expression datasets of the periodic genes, we constructed gene networks and found that the networks represent typical scale-free structure. Our findings demonstrate that the diurnal changes in the transcriptome in B. distachyon leaves have a sparse network structure, demonstrating the spatiotemporal gene regulatory network over the cyclic phase transitions in B. distachyon diurnal growth.


Postoperative analgesics in South African Clawed frogs (Xenopus laevis) after surgical harvest of oocytes.
Heightened awareness for the welfare of earlier-evolved laboratory species has prompted increasing inquiries by institutional animal care committees, investigators, and laboratory animal veterinarians regarding the need for post-surgical analgesics in laboratory Xenopus. Basic research into the mechanisms and regulation of pain in Rana pipiens has demonstrated the clinical potential of opioid, alpha2-adrenergic, and non-opioid analgesic agents in amphibians. However, clinical studies using objectively established indices of amphibian pain, or pharmacological studies in either Rana pipiens or laboratory Xenopus have not been conducted. As discussed above, comparison of limited lethality data suggests that the safety index for these agents is quite narrow in Rana pipiens. Analgesic use in laboratory Xenopus has the added risk of drowning due to over sedation. Drug doses extrapolated from such studies and intended to provide pain relief in Xenopus should therefore be considered very carefully. An additional concern for laboratory Xenopus is that the effects of these agents on amphibian oogenesis, oocyte quality, and embryogenesis are unknown. As the numbers of laboratory Xenopus used in basic and biomedical research continues to increase, clinical studies that address all of these issues cannot come too soon.


Predicting 195 Pt NMR Chemical Shifts in Water-Soluble Inorganic/Organometallic Complexes with a Fast and Simple Protocol Combining Semiempirical Modeling and Machine Learning.
Water-soluble Pt complexes are the key components in medicinal chemistry and catalysis. The well-known cisplatin family of anticancer drugs and industrial hydrosylilation catalysts are two leading examples. On the molecular level, the activity mechanisms of such complexes mostly involve changes in the Pt coordination sphere. Using 195 Pt NMR spectroscopy for operando monitoring would be a valuable tool for uncovering the activity mechanisms; however, reliable approaches for the rapid correlation of Pt complex structure with 195 Pt chemical shifts are very challenging and not available for everyday research practice. While NMR shielding is a response property, molecular 3D structure determines NMR spectra, as widely known, which allows us to build up 3D structure to 195 Pt chemical shift correlations. Accordingly, we present a new workflow for the determination of lowest-energy configurational/conformational isomers based on the GFN2-xTB semiempirical method and prediction of corresponding chemical shifts with a Machine Learning (ML) model tuned for Pt complexes. The workflow was designed for the prediction of 195 Pt chemical shifts of water-soluble Pt(II) and Pt(IV) anionic, neutral, and cationic complexes with halide, NO2 - , (di)amino, and (di)carboxylate ligands with chemical shift values ranging from -6293 to 7090 ppm. The model offered an accuracy (normalized root-mean-square deviation/RMSD) of 1.08 %/145.02 ppm on the held-out test set.


Human resistin promotes neutrophil proinflammatory activation and neutrophil extracellular trap formation and increases severity of acute lung injury.
Although resistin was recently found to modulate insulin resistance in preclinical models of type II diabetes and obesity, recent studies also suggested that resistin has proinflammatory properties. We examined whether the human-specific variant of resistin affects neutrophil activation and the severity of LPS-induced acute lung injury. Because human and mouse resistin have distinct patterns of tissue distribution, experiments were performed using humanized resistin mice that exclusively express human resistin (hRTN(+/-)(/-)) but are deficient in mouse resistin. Enhanced production of TNF-α or MIP-2 was found in LPS-treated hRtn(+/-/-) neutrophils compared with control Rtn(-/-/-) neutrophils. Expression of human resistin inhibited the activation of AMP-activated protein kinase, a major sensor and regulator of cellular bioenergetics that also is implicated in inhibiting inflammatory activity of neutrophils and macrophages. In addition to the ability of resistin to sensitize neutrophils to LPS stimulation, human resistin enhanced neutrophil extracellular trap formation. In LPS-induced acute lung injury, humanized resistin mice demonstrated enhanced production of proinflammatory cytokines, more severe pulmonary edema, increased neutrophil extracellular trap formation, and elevated concentration of the alarmins HMGB1 and histone 3 in the lungs. Our results suggest that human resistin may play an important contributory role in enhancing TLR4-induced inflammatory responses, and it may be a target for future therapies aimed at reducing the severity of acute lung injury and other inflammatory situations in which neutrophils play a major role.


Localized compressional velocity decrease precursory to the kalapana, hawaii, earthquake.
A delay in the arrival times of compressional or P waves of 0.15 to 0.2 second from deep distant earthquakes has been detected at the closest seismograph station to the 20 November 1975 earthquake at Kalapana, Hawaii (surface-wave magnitude MS = 7.2). This delay appeared approximately 3.5 years prior to the quake, and travel times returned to normal several months before it. The P-wave arrival times at other nearby stations remained constant during this period, an indication that the decreased velocity implied by the delay in travel time was associated with this normal-faulting earthquake and was confined to distances less than 20 kilometers from the epicenter.


Increased Vacuolar ATPase Activity Correlated With CAM Induction in Mesembryanthemum crystallinum and Kalanchoë blossfeldiana cv. Tom Thumb.
Vacuolar ATPase activities were determined by differential inhibition of homogenates of isolated protoplasts (using the inhibitors molybdate for acid phosphatases, vanadate for plasmalemma ATPase, azide for mitochondrial ATPase, and phlorizin for chloroplast ATPase) and in preparations of isolated vacuoles of Mesembryanthemum crystallinum and Kalanchoë blossfeldiana cv. Tom Thumb. Crassulacean acid metabolism (CAM) was induced in M. crystallinum by NaCl-salinity and in K. blossfeldiana by short-day treatments. Vacuolar ATPase activities increased several-fold during the transition from C(3) photosynthesis to CAM. The increase was quantitatively related to the rates of nocturnal maliacid accumulation in CAM assuming a stoichiometry of 2 H(+) pumped into the vacuole for 1 ATP hydrolyzed and 1 malate(2-) anion transported by secondary flux coupling. In M. crystallinum increased vacuolar ATPase activities were truly correlated with the degree of CAM expressed and not with NaCl accumulation due to the salinity treatment. Some properties of the vacuolar A TPase of M. crystallinum characterized in vacuole preparations were a pH-optimum near 8.0, an apparent K(m) (MgATP(2-)) of 0.20 to 0.29 mM, and an approximately 70 % inhibition by 50 mM nitrate.


Risk Factors and Current Health-Seeking Patterns of Migrants in Northeastern Mexico: Healthcare Needs for a Socially Vulnerable Population.
This study identified risk factors for health and access to healthcare services of migrants during their journey across Mexico to the United States. Data were collected in shelters located in Monterrey, the largest city of northeastern Mexico, through a basic clinical examination and a survey completed by 75 migrants; 92% of them were undocumented Central Americans. During their transit, they are at a high risk of contracting, developing, and transmitting diseases. The need of working to survive affects health-seeking behavior and a constant fear of being traced keeps migrants away from public health services, which delays diagnosis and treatment of diseases. Negligent lifestyles, such as smoking, drinking (31.8% of men and 11.1% of women), and drug abuse (13% of men and 11% of women), were found. Regarding tuberculosis (TB), undocumented migrants are usually not screened, even though they come from countries with a high TB burden. Besides, they might be overexposed to TB because of their living conditions in overcrowded places with deficient hygiene, protection, and malnutrition (54.7% of the sample). Possible comorbidities like acquired immune deficiency syndrome (AIDS; 4%) and diabetes (2.7%, but probably under-diagnosed) were referred. Migrants have little TB knowledge, which is independent of their level of education or a previous experience of deportation. About one-third of the migrants were totally unfamiliar with TB-related symptoms, while 36% had correct knowledge of basic TB symptoms. We conclude that a shortage of information on the highly vulnerable migratory population combined with a lack of social support and health education among migrants may play a significant role in the spread of communicable diseases. We recommend that health authorities address this urgent, binational, public health concern in order to prevent outbreaks of emerging infections.


Associations Between Neighborhood Problems and Sexual Behaviors Among Black Men Who Have Sex with Men in the Deep South: The MARI Study.
There is a disproportionately high HIV incidence among Black men who have sex with men (MSM) despite equal or lower levels of HIV risk behaviors compared to White MSM. Due to high levels of racial segregation in the U.S., Black MSM have an elevated likelihood of living in neighborhoods that contain psychosocial stressors, which, in turn, may increase behaviors promoting HIV infection. We examined associations between perceived neighborhood problems and sexual behaviors among Black MSM in the Deep South, a population at highest risk of HIV. Data came from the MARI Study, which included Black MSM ages 18-66 years recruited from the Jackson, MS, and Atlanta, GA, metropolitan areas (n = 377). Participants completed questions about neighborhood problems (e.g., excessive noise, heavy traffic/speeding cars and trash/litter) and sexual behaviors (e.g., condomless sex and drug use before or during sex). We used Poisson's regression model with robust standard errors to estimate the adjusted prevalence ratio (aPR; 95% confidence intervals [CI]) of neighborhood problems (coded as tertiles [tertile 1 = low neighborhood problems, tertile 2 = medium neighborhood problems, tertile 3 = high neighborhood problems] as well as continuously) with sexual behaviors, after adjustment for sociodemographic characteristics and other variables. About one-fourth of the sample reported at least one neighborhood problem, with the most common (31.6%) being no/poorly maintained sidewalks, which indicates an infrastructural problem. In multivariable models, compared to those in the lowest tertile, those reporting more neighborhood problems (tertile 2: aPR = 1.49, 95% CI = 1.04, 2.14 and tertile 3: aPR = 1.53, 95% CI = 1.05, 2.24) reported more drug use before or during sex (p for trend = .027). Neighborhood problems may promote behaviors (e.g., drug use before or during sex) conducive to HIV infection. Structural interventions could improve community infrastructure to reduce neighborhood problems (e.g., no/poorly maintained sidewalks and litter). These interventions may help to reduce HIV incidence among Black MSM in the Deep South.


MACC1 regulates clathrin-mediated endocytosis and receptor recycling of transferrin receptor and EGFR in colorectal cancer.
Metastasis Associated in Colon Cancer 1 (MACC1) is a novel prognostic, predictive and causal biomarker for tumor progression and metastasis in many cancer types, including colorectal cancer. Besides its clinical value, little is known about its molecular function. Its similarity to SH3BP4, involved in regulating uptake and recycling of transmembrane receptors, suggests a role of MACC1 in endocytosis. By exploring the MACC1 interactome, we identified the clathrin-mediated endocytosis (CME)-associated proteins CLTC, DNM2 and AP-2 as MACC1 binding partners. We unveiled a MACC1-dependent routing of internalized transferrin receptor towards recycling. Elevated MACC1 expression caused also the activation and internalization of EGFR, a higher rate of receptor recycling, as well as earlier and stronger receptor activation and downstream signaling. These effects are limited by deletion of CME-related protein interaction sites in MACC1. Thus, MACC1 regulates CME and receptor recycling, causing increased growth factor-mediated downstream signaling and cell proliferation. This novel mechanism unveils potential therapeutic intervention points restricting MACC1-driven metastasis.


Genomic signatures predict migration and spawning failure in wild Canadian salmon.
Long-term population viability of Fraser River sockeye salmon (Oncorhynchus nerka) is threatened by unusually high levels of mortality as they swim to their spawning areas before they spawn. Functional genomic studies on biopsied gill tissue from tagged wild adults that were tracked through ocean and river environments revealed physiological profiles predictive of successful migration and spawning. We identified a common genomic profile that was correlated with survival in each study. In ocean-tagged fish, a mortality-related genomic signature was associated with a 13.5-fold greater chance of dying en route. In river-tagged fish, the same genomic signature was associated with a 50% increase in mortality before reaching the spawning grounds in one of three stocks tested. At the spawning grounds, the same signature was associated with 3.7-fold greater odds of dying without spawning. Functional analysis raises the possibility that the mortality-related signature reflects a viral infection.


A morphometric study on flexor carpi radialis muscle of the forearm: A cadaveric study.
Flexor carpi radialis (FCR) is frequently used in reconstruction procedures involving the hand and wrist regions. Despite having tremendous clinical significance, very few studies have been conducted on the heterogeneity of the FCR. Its surgical importance in grafting techniques and tendon transfers prompted us to study the morphometry of FCR. In this study, we used 37 upper limbs (25 right and 12 left sides) of formalin embalmed cadavers. FCR was neatly dissected in all upper limbs to measure its total length, muscle belly length, tendon length, width of muscle belly at proximal and distal ends, width of tendon at proximal and distal ends. Average total length of the muscle, muscle belly length, tendon length, width of muscle belly at the proximal end, width of muscle belly at the distal end, width of tendon at the proximal end, width of tendon at the distal end were 27.1 ± 2.13, 12.38 ± 2.23, 14.99 ± 2.36, 1.99 ± 0.72, 1.59 ± 0.48, 1.59 ± 0.48 and 0.42 ± 0.12 cm, respectively. Ratio of mean muscle length and tendon length, ratio of mean tendon length and total length were found to be 0.45 ± 0.07 and 0.55 ± 0.07, respectively. No noted variations were observed in the proximal and distal attachments of the FCR muscle. Morphometric data of FCR reported increases the existing knowledge on the heterogeneity of FCR morphology, and further forms an anatomic basis while designing/elevating the graft in certain surgical procedures in hand and wrist regions.


Profile of histone lysine methylation across transcribed mammalian chromatin.
Complex patterns of histone lysine methylation encode distinct functions within chromatin. We previously reported that trimethylation of lysine 9 of histone H3 (H3K9) occurs at both silent heterochromatin and at the transcribed regions of active mammalian genes, suggesting that the extent of histone lysine methylation involved in mammalian gene activation is not completely defined. To identify additional sites of histone methylation that respond to mammalian gene activity, we describe here a comparative assessment of all six known positions of histone lysine methylation and relate them to gene transcription. Using several model loci, we observed high trimethylation of H3K4, H3K9, H3K36, and H3K79 in the transcribed region, consistent with previous findings. We identify H4K20 monomethylation, a modification previously linked with repression, as a mark of transcription elongation in mammalian cells. In contrast, H3K27 monomethylation, a modification enriched at pericentromeric heterochromatin, was observed broadly distributed throughout all euchromatic sites analyzed, with selective depletion in the vicinity of the transcription start sites at active genes. Together, these results underscore that similar to other described methyl-lysine modifications, H4K20 and H3K27 monomethylation are versatile and dynamic with respect to gene activity, suggesting the existence of novel site-specific methyltransferases and demethylases coupled to the transcription cycle.


Parkinsonism in neuroleptic-naive schizophrenic patients.
Previous studies in schizophrenia have identified abnormalities involving the basal ganglia, but the contribution of neuroleptics to the motor system abnormalities in schizophrenia is usually a confounding factor. This study addressed the issue of whether parkinsonism, a reflection of dopaminergic hypofunction, occurs in schizophrenia per se. Clinical ratings and quantitative instrumental measures of parkinsonian rigidity, tremor, and bradykinesia were obtained in 24 neuroleptic-naive schizophrenic patients and 24 age- and gender-matched comparison subjects. According to the clinical ratings, 21% of the schizophrenic patients had rigidity and 12% had bradykinesia, in contrast to none of the normal comparison subjects. With the use of instrumental measures, rigidity and tremor were observed in 29% and 37%, respectively, of the schizophrenic patients, compared to 4% and none in the normal comparison group. The schizophrenic patients also exhibited greater right-side than left-side parkinsonism. The findings suggest that extrapyramidal motor signs may be part of schizophrenia proper and that some patients with schizophrenia have left striatal hypodopaminergia unrelated to neuroleptic treatment.


Electrophysiological and morphological properties of layers V-VI principal pyramidal cells in rat prefrontal cortex in vitro.
This study examined the electrophysiological and morphological characteristics of layers V-VI pyramidal prefrontal cortex (PFC) neurons. In vitro intracellular recordings coupled with biocytin injections that preserved some of the PFC efferents to the nucleus accumbens (NAc) were made in brain slices. Four principal pyramidal cell types were identified and classified as regular spiking (RS) (19%), intrinsic bursting (IB) (64%), repetitive oscillatory bursting (ROB) (13%), and intermediate (IM) (4%) types. All PFC cells exhibited either subthreshold oscillation in membrane voltage or pacemaker-like rhythmic firing. IB neurons were demonstrated electrophysiologically and cytochemically to be PFC-->NAc neurons. In all IB and some RS neurons, a tetrodotoxin-sensitive, slowly inactivating Na+ current and a transient Ni(2+)-sensitive, low-threshold Ca2+ current mediated subthreshold inward rectification. During sustained membrane depolarization, the Na+ current was opposed by a 4-aminopyridine-sensitive, outwardly rectifying, slowly inactivating K+ current. Together, these three currents controlled the firing threshold of the PFC neurons. All IB and ROB cells also had postspike Ca(2+)-mediated depolarizing afterpotentials, postburst Ca(2+)-dependent after hyperpolarizations, and low- and high-threshold Ca2+ spikes. In addition, ROB cells had a hyperpolarizing "sag" mediated by the cationic conductance, Ih. IB and ROB neurons had extensive dendritic trees and radially ascending or tangentially projecting axon collaterals. RS and IM cells had comparatively simpler morphological profiles. These electrophysiological and morphological properties of the four principal pyramidal PFC cell types have provided valuable details for understanding further how PFC processes input and transmit outputs to regions such as the NAc.


Necrotizing pancreatitis after transcatheter arterial chemoembolization for hepatocellular carcinoma.
A patient who developed necrotizing pancreatitis after transcatheter arterial chemoembolization (TACE) for hepatocellular carcinoma (HCC) is presented. A 55- year-old man had been followed for chronic hepatitis B infection for 10 years at another institution. He presented with multiple masses in the right lobe of the liver and a metastasis in the left adrenal gland. He was referred after a percutaneous liver biopsy which revealed a moderately differentiated HCC. He was treated by TACE. At the third session of TACE, the right hepatic artery was found to be thrombosed; however, angiography also demonstrated collateral feeder vessels (arising from the pancreaticoduodenal artery) which were used for treatment. He developed necrotizing pancreatitis, possibly due to regurgitation of the chemotherapeutic agents to the pancreas. He recovered without complications with imipenem-cilastatin prophylaxis. Acute pancreatitis is a rare but severe complication of TACE. Selective catheterization of the tumor vessels is the established standard in TACE. A careful risk-benefit analysis is mandatory in patients with abnormal collateral vessels. Treatment of acute necrotizing pancreatitis (ANP) after TACE is the same as the accepted approach to ANP due to other causes.


Functional double-shelled silicon nanocrystals for two-photon fluorescence cell imaging: spectral evolution and tuning.
Functional near-IR (NIR) emitting nanoparticles (NPs) adapted for two-photon excitation fluorescence cell imaging were obtained starting from octadecyl-terminated silicon nanocrystals (ncSi-OD) of narrow photoluminescence (PL) spectra having no long emission tails, continuously tunable over the 700-1000 nm window, PL quantum yields exceeding 30%, and PL lifetimes of 300 μs or longer. These NPs, consisting of a Pluronic F127 shell and a core made up of assembled ncSi-OD kept apart by an octadecyl (OD) layer, were readily internalized into the cytosol, but not the nucleus, of NIH3T3 cells and were non-toxic. Asymmetrical field-flow fractionation (AF4) analysis was carried out to determine the size of the NPs in water. HiLyte Fluor 750 amine was linked via an amide link to NPs prepared with Pluronic-F127-COOH, as a first demonstration of functional NIR-emitting water dispersible ncSi-based nanoparticles.


Early failure of the Ultima metal-on-metal total hip replacement in the presence of normal plain radiographs.
Metal-on-metal total hip replacement has been targeted at younger patients with anticipated long-term survival, but the effect of the production of metal ions is a concern because of their possible toxicity to cells. We have reviewed the results of the use of the Ultima hybrid metal-on-metal total hip replacement, with a cemented polished tapered femoral component with a 28 mm diameter and a cobalt-chrome (CoCr) modular head, articulating with a 28 mm CoCr acetabular bearing surface secured in a titanium alloy uncemented shell. Between 1997 and 2004, 545 patients with 652 affected hips underwent replacement using this system. Up to 31 January 2008, 90 (13.8%) hips in 82 patients had been revised. Pain was the sole reason for revision in 44 hips (48.9%) of which 35 had normal plain radiographs. Peri-prosthetic fractures occurred in 17 hips (18.9%) with early dislocation in three (3.3%) and late dislocation in 16 (17.8%). Infection was found in nine hips (10.0%). At operation, a range of changes was noted including cavities containing cloudy fluid under pressure, necrotic soft tissues with avulsed tendons and denuded osteonecrotic upper femora. Corrosion was frequently observed on the retrieved cemented part of the femoral component. Typically, the peri-operative findings confirmed those found on pre-operative metal artefact reduction sequence MRI and histological examination showed severe necrosis. Metal artefact reduction sequence MRI proved to be useful when investigating these patients with pain in the absence of adverse plain radiological features.


Detecting MMN in Infants EEG with Singular Value Decomposition.
Mismatch negativity (MMN) is an EEG voltage fluctuation caused by the brain's automatic reaction to unexpected changes in a repetitive stimulation. In an experiment we studied 68 infants of which 2/3 were born preterm. Due to noise of large amplitude, the MMN is difficult to detect in a single infant's EEG. Therefore grand average, which is a average of many subjects EEG recordings, is sometimes used. In this paper singular value decomposition (SVD) is proposed as an alternative to grand average. Consider the SVD USigmaVT = M, where the rows of M contains noisy EEG epochs. Usually data is projected onto the leftmost column of V since this column represent the largest common component of the rows of M. When data is affected by noise of a very large amplitude we may need to choose another column of V. In this paper we propose to choose the leftmost column of V such that the elements of the corresponding column of U has approximately equal values.


stg fimbrial operon from S. Typhi STH2370 contributes to association and cell disruption of epithelial and macrophage-like cells.
Salmonella enterica serovar Typhi (S. Typhi) stg operon, encoding a chaperone/usher fimbria (CU), contributes to an increased adherence to human epithelial cells. However, one report suggests that the presence of the Stg fimbria impairs the monocyte--bacteria association, as deduced by the lower level of invasion to macrophage-like cells observed when the stg fimbrial cluster was overexpressed. Nevertheless, since other CU fimbrial structures increase the entry of S. Typhi into macrophages, and considering that transcriptomic analyses revealed that stg operon is indeed expressed in macrophages, we reassessed the role of the stg operon in the interaction between S. Typhi strain STH2370 and human cells, including macrophage-like cells and mononuclear cells directly taken from human peripheral blood. We compared S. Typhi STH2370 WT, a Chilean clinical strain, and the S. Typhi STH2370 Δstg mutant with respect to association and invasion using epithelial and macrophage-like cells. We observed that deletion of stg operon reduced the association and invasion of S. Typhi, in both cellular types. The presence of the cloned stg operon restored the WT phenotype in all the cases. Moreover, we compared Salmonella enterica sv. Typhimurium 14028s (S. Typhimurium, a serovar lacking stg operon) and S. Typhimurium heterologously expressing S. Typhi stg. We found that the latter presents an increased cell disruption of polarized epithelial cells and an increased association in both epithelial and macrophage-like cells. S. Typhi stg operon encodes a functional adhesin that participates in the interaction bacteria-eukaryotic cells, including epithelial cells and macrophages-like cells. The phenotypes associated to stg operon include increased association and consequent invasion in bacteria-eukaryotic cells, and cell disruption.


Outcomes of transmetatarsal amputations in patients with diabetes mellitus.
The purpose of this study was to report on the long-term outcomes of transmetatarsal amputations secondary to sequelae of diabetes mellitus. We abstracted data from 35 diabetic patients receiving a transmetatarsal amputation over a 6-month period in 1992. Patients were followed for a mean 15.1 +/- 10.1 months. The results indicated that the most predictive factor determining higher level amputation (transfemoral or transtibial) appeared to be the actual indication for surgery (90.0% ischemia versus 4.0% infection, chi 2 = 21.7, odds ratio = 220, 95% confidence interval = 12.5-3885.0, p < 0.05). Those with a diagnosis of infection without underlying critical ischemia were significantly more likely to heal at the level of the foot. While all patients presenting for care had dramatically impaired nutritional values and elevated glucose, albumin was significantly lower in subjects receiving a transfemoral or transtibial revision. High-level amputees were also significantly less likely to have been prescribed depth-inlay shoe gear prior to their amputation (48.0% vs. 10.0%, chi 2 = 4.4, odds ratio = 8.3, 95% confidence interval = 1.0-75.7, p < 0.05). Those with a diagnosis of infection without underlying critical ischemia were significantly more likely to heal at the level of the foot. Though revision rates are high, the success rates are also high if that is defined as retaining the foot and providing a prosthesis-free normal gait.


Neurogenic pulmonary oedema in a 13-year-old boy in the course of symptomatic epilepsy--case report.
Pulmonary oedema with severe, dramatic course following CNS injury was termed neurogenic pulmonary oedema (NPO). NPO was mainly described as a consequence of grand mal seizures, subarachnoid bleeding, intracranial bleeding or head injury. However, the pathogenesis of NPO is not entirely clear yet. In the majority of cases, early or classic symptoms of pulmonary oedema are evident from several minutes up to several hours after CNS damage. Dyspnoea, chest pain, bloody expectoration are observed shortly after consciousness disorders, although NPO may occasionally be diagnosed on the basis of chest x-ray in patients with no clinical symptoms. Tachypnoea, tachycardia, rales without any changes in cardiac system are usually observed during physical examination. The ailments withdraw quickly in the majority of patients, who may require oxygen therapy at most. NPO has been well-known in adults, but our knowledge of its occurrence in children is still rather sparse. The current work presents a case of a 13-year-old boy with pulmonary oedema as a post-seizure complication.


Evaluation of a portable clinical analyzer for the determination of blood gas partial pressures, electrolyte concentrations, and hematocrit in venous blood samples collected from cattle, horses, and sheep.
To compare results reported for blood gas partial pressures, electrolyte concentrations, and Hct in venous blood samples collected from cattle, horses, and sheep and analyzed by use of a portable clinical analyzer (PCA) and reference analyzer (RA). Clinically normal animals (24 cattle, 22 horses, and 22 sheep). pH; Pco(2); Po(2); total carbon dioxide concentration; oxygen saturation; base excess; concentrations of HCO(3)(-), Na(+), K(+), and ionized calcium; Hct; and hemoglobin concentration were determined with a PCA. Results were compared with those obtained for the same blood sample with an RA. Bias (mean difference) and variability (95% confidence interval) were determined for all data reported. Data were also subjected to analyses by Deming regression and Pearson correlation. Analysis of Bland-Altman plots revealed good agreement between results obtained with the PCA and those obtained with the RA for pH and total carbon dioxide concentration in cattle, K(+) concentration in horses and sheep, and base excess in horses. Except for Na(+) concentration and Hct in horses and sheep, correlation was good or excellent for most variables reported. Data from blood gas and electrolyte analyses obtained by use of the PCA can be used to evaluate the health status of cattle, horses, and sheep. Furthermore, the handheld PCA device may have a great advantage over the RA device as a result of the ability to analyze blood samples on farms that may be located far from urban centers.


Schizophrenia relapse and the clinical usefulness of once-monthly aripiprazole depot injection.
Improving medication adherence is critical to improving outcomes in patients with schizophrenia. A long-acting injectable (depot) antipsychotic is one of the most effective methods for improving treatment adherence and decreasing rehospitalization rates in patients with schizophrenia. Until recently, only three second-generation antipsychotics were available in a long-acting injectable formulation (risperidone, paliperidone, and olanzapine). In this respect, the emergence of long-acting aripiprazole injection (ALAI), approved by the US Food and Drug Administration for the treatment of schizophrenia in 2013, is timely. ALAI is a lyophilized powder of aripiprazole, and the aripiprazole molecule is unmodified. The initial and target dosage of ALAI is 400 mg once monthly, but it could be reduced to 300 mg if adverse reactions occur with 400 mg. When first administering ALAI, it is recommended to continue treatment with oral aripiprazole (10-20 mg/day) or another oral antipsychotic for 2 weeks in order to maintain therapeutic antipsychotic concentrations. The primary clearance route for ALAI is hepatic, ie, cytochrome P450 (CYP)2D6 and CYP3A4, so dose adjustment is required in poor CYP2D6 metabolizers. The efficacy of ALAI was demonstrated in three studies. A randomized controlled trial that formed the basis for approval of ALAI in the treatment of schizophrenia showed that ALAI significantly delayed time to impending relapse when compared with placebo (P<0.0001, log-rank test). An open-label, mirror study demonstrated that total psychiatric hospitalization rates were significantly lower after switching from oral antipsychotics to ALAI. Another randomized controlled trial presented in poster form suggested that ALAI 400 mg was comparable with oral aripiprazole 10-30 mg in preventing relapse. ALAI was generally well tolerated during both short-term and long-term studies. Its tolerability profile, including extrapyramidal symptoms and clinically relevant metabolic parameters, was similar to placebo. However, insomnia, headache, anxiety, akathisia, weight gain, injection site pain, and tremor need clinical attention. These studies suggest that ALAI is a viable treatment option for patients with schizophrenia, but direct head-to-head comparisons between ALAI and other long-acting injectable antipsychotics are needed to elucidate its risk-benefit profile.


3D printing of rat salivary glands: The submandibular-sublingual complex.
The morphology and the functionality of the murid glandular complex, composed of the submandibular and sublingual salivary glands (SSC), were the object of several studies conducted mainly using magnetic resonance imaging (MRI). Using a 4.7 T scanner and a manganese-based contrast agent, we improved the signal-to-noise ratio of the SSC relating to the surrounding anatomical structures allowing to obtain high-contrast 3D images of the SSC. In the last few years, the large development in resin melting techniques opened the way for printing 3D objects starting from a 3D stack of images. Here, we demonstrate the feasibility of the 3D printing technique of soft tissues such as the SSC in the rat with the aim to improve the visualization of the organs. This approach is useful to preserve the real in vivo morphology of the SCC in living animals avoiding the anatomical shape changes due to the lack of relationships with the surrounding organs in case of extraction. It is also harmless, repeatable and can be applied to explore volumetric changes occurring during body growth, excretory duct obstruction, tumorigenesis and regeneration processes. 3D printing allows to obtain a solid object with the same shape of the organ of interest, which can be observed, freely rotated and manipulated. To increase the visibility of the details, it is possible to print the organs with a selected zoom factor, useful as in case of tiny organs in small mammalia. An immediate application of this technique is represented by educational classes.


Inflammatory disease protective R381Q IL23 receptor polymorphism results in decreased primary CD4+ and CD8+ human T-cell functional responses.
The SNP (c.1142G > A;p.R381Q) in the IL-23 receptor (IL23R) confers protection from multiple inflammatory diseases, representing one of the most significant human genetic polymorphisms in autoimmunity. We, therefore, sought to define the functional consequences of this clinically significant variant. We find that CD4+CD45RO+ and CD8+ T cells from healthy R381Q IL23R carriers show decreased IL-23-dependent IL-17 and IL-22 production relative to WT IL23R individuals. This was associated with a lower percentage of circulating Th17 and Tc17 cells. Furthermore, CD8+ T cells from R381Q IL23R individuals showed decreased IL-23-dependent expansion and signal transducer and activator of transcription 3 (STAT3) activation compared with WT CD8+ T cells. Importantly, cells transfected with the IL23R glutamine variant show decreased IL-23-mediated signaling compared with the IL23R arginine allele. Our results show that the R381Q IL23R variant leads to selective, potentially desirable, loss of function alterations in primary human CD4+ and CD8+ T cells, resulting in highly significant protection against autoimmunity.