Thrombin is the major serum factor stimulating phosphoinositide turnover, but not DNA synthesis in human neuroblastoma SH-EP cells.
Fetal calf serum stimulates both phosphoinositide turnover and DNA synthesis in SH-EP cells. The phosphoinositide turnover-stimulating activity of serum is largely (70%) reduced in the presence of hirudin, a blocker of thrombin activity. Yet, hirudin does not alter the ability of serum to stimulate DNA synthesis. Purified alpha-thrombin is a potent (EC50, 35 pM) stimulator of phosphoinositide turnover in SH-EP cells, but induces DNA synthesis only at much higher concentrations (10 nM-1 microM). Thus, serum thrombin accounts for most of the ability of serum to stimulate phosphoinositide hydrolysis, but not for the effect of serum on cell division, since the concentration of thrombin in serum is not sufficient to induce DNA synthesis. These data suggest that hydrolysis of inositol lipids may not be the main signalling event mediating the mitogenic effects of alpha-thrombin.


Recombinant human interferon alpha-2a therapy in children with chronic immune thrombocytopenic purpura.
In a prospective study, 11 children with chronic immune thrombocytopenic purpura between ages 3 and 18 years were treated with recombinant human interferon alpha 2a (rhIFN alpha-2a). A dose of 3 x 10(6) U/m2 three times weekly for 4 to 5 weeks (one cycle) was administered. Patients were treated with one to four cycles of rhIFN alpha-2a, and the outcomes were measured initially and 18 to 30 months after the last cycle. Good therapeutic responses (defined as platelet count >100 x 10(9)/L) lasting for 18 to 30 months from the last interferon cycle were achieved in 6 of the 11 (55%) patients, including one with a probable spontaneous remission. Fair responses (platelet count 31-60 x 10(9)/L) for 18 months were achieved in 3 of the 11 (27%) patients. Only two patients, each treated only with one interferon cycle, exhibited no response. Side effects of treatment included fever and a flu-like syndrome, which were usually present during the first 14 days of therapy only. Interferon-alpha appears to be an effective therapeutic approach to children with chronic immune thrombocytopenic purpura, with the potential of sustained long-term remission. A randomized, placebo-controlled study is needed to confirm its role in this population.


Rhodium-catalyzed domino heterocyclization and [(3+2)+2] carbocyclization: construction of fused tricycloheptadienes.
A rhodium-catalyzed domino heterocyclization and formal [(3+2)+2] carbocyclization reaction of readily available diyne-enones and alkynes was developed.


Purification and cloning of glyoxalase II from rat liver.
Glyoxalase (GLO) II, which is a component of GLO system and catalyze the conversion of S-lactoyl-glutathione to D-lactate, was purified 1488 fold from rat liver by two steps of Affigel blue and carbobenzoxyglutathione-Sepharose 4B affinity chromatography. The molecular weight of the enzyme was estimated to be 29 kDa which is similar to those from other species. The sequence of N-terminal 9 amino acid residues was determined to be MGIRLLPAT. This was then used to synthesize degenerative primers. cDNA clone was isolated by first synthesizing cDNA from RNA and then PCR amplification. The sequence of cDNA clone was determined by serial sequencing analysis.


Silver nanoparticle-coated suture effectively reduces inflammation and improves mechanical strength at intestinal anastomosis in mice.
Our previous studies have revealed that silver nanoparticles (AgNPs) had anti-inflammatory properties. In this study, we coated AgNPs onto the surface of absorbable suture, to further explore their anti-inflammatory efficacy and potential clinical application using an intestinal anastomosis model. Layer-by-layer deposition was used to coat AgNPs on absorbable sutures. Scanning electron microscopy (SEM) was conducted to observe the morphology and distribution of AgNPs on suture surface. 1cm of either non-coated suture, suture coated with antibiotics or AgNPs-coated suture was placed on E. coli overlay of LB agar plates to test for bacterial inhibition. The respective sutures were then used for ileal anastomosis in mice. The anastomotic sites were harvested to investigate the degree of tissue inflammation and cell proliferation, as well as collagen deposition. Furthermore, burst pressure measurement was employed to test for mechanical properties. SEM observation indicated AgNPs could be immobilized and distributed on suture surface evenly. AgNPs-coated suture had the best in vitro anti-bacterial efficacy when compared with other groups. Subsequent immunohistochemistry in the intestinal anastomosis model showed significantly less inflammatory cell infiltration (macrophage and neutrophil) and better collagen deposition in the anastomotic tissue in the AgNPs-coated suture group. Burst pressure measurement in healed anastomosis further confirmed that AgNPs-coated suture had better mechanical properties. Our study suggests that AgNPs-coated sutures can improve anastomosis healing due to better mechanical properties from reduced inflammation.


The neural basis of one's own conscious and unconscious emotional states.
The study of emotional states has recently received considerable attention within the cognitive and neural sciences. However, limited work has been done to synthesize this growing body of literature within a coherent hierarchical, neuro-cognitive framework. In this article, we review evidence pertaining to three interacting hierarchical neural systems associated with the generation, perception and regulation of one's own emotional state. In the framework we propose, emotion generation proceeds through a series of appraisal mechanisms - some of which appear to require more cognitively sophisticated computational processing (and hence more time) than others - that ultimately trigger iterative adjustments to one's bodily state (as well as to the modes of processing in other cognitive systems). Perceiving one's own emotions then involves a multi-stage interoceptive/somatosensory process by which these body state patterns are detected and assigned conceptual emotional meaning. Finally, emotion regulation can be understood as a hierarchical control system that, at various levels, modulates autonomic reactions, appraisal mechanisms, attention, the contents of working memory, and goal-directed action selection. We highlight implications this integrative model may have for competing theories of emotion and emotional consciousness and for guiding future research.


Monitoring HIV Testing in the United States: Consequences of Methodology Changes to National Surveys.
In 2011, the National Health Interview Survey (NHIS), an in-person household interview, revised the human immunodeficiency virus (HIV) section of the survey and the Behavioral Risk Factor Surveillance System (BRFSS), a telephone-based survey, added cellphone numbers to its sampling frame. We sought to determine how these changes might affect assessment of HIV testing trends. We used linear regression with pairwise contrasts with 2003-2013 data from NHIS and BRFSS to compare percentages of persons aged 18-64 years who reported HIV testing in landline versus cellphone-only households before and after 2011, when NHIS revised its in-person questionnaire and BRFSS added cellphone numbers to its telephone-based sample. In NHIS, the percentage of persons in cellphone-only households increased 13-fold from 2003 to 2013. The percentage ever tested for HIV was 6%-10% higher among persons in cellphone-only than landline households. The percentage ever tested for HIV increased significantly from 40.2% in 2003 to 45.0% in 2010, but was significantly lower in 2011 (40.6%) and 2012 (39.7%). In BRFSS, the percentage ever tested decreased significantly from 45.9% in 2003 to 40.2% in 2010, but increased to 42.9% in 2011 and 43.5% in 2013. HIV testing estimates were lower after NHIS questionnaire changes but higher after BRFSS methodology changes. Data before and after 2011 are not comparable, complicating assessment of trends.


Effects of bilateral early breast sucking and unilateral early breast sucking within 2 h after delivery on lactation, breast distending pain and postpartum lochia.
To investigate the effects of bilateral early breast sucking and unilateral early breast sucking within 2 h after delivery on lactation, breast distending pain and postpartum lochia. The clinical data of 128 parturients were analyzed retrospectively. According to the different ways of early breast suction, the parturients were divided into control group (n=64) and observation group (n=64). Among them, unilateral early sucking was performed within 2 h after delivery in the control group, while bilateral early sucking was carried out in the observation group. We compared the lactation, breast distending pain, postpartum lochia, nutritional status, coagulation index, complications and breast feeding success rate. The lactation amount of the observation group was higher than that of the control group on the 3rd, 5th and 7th day after delivery; the VAS scores showed opposite trends; the amount of lochia in the observation group was less than that in the control group; the ALb, PA and Hb levels in the observation group were higher on the 7th day after delivery. Compared with the control group, the FIB, PT and APTT time was shorter and the D-D level was higher; the incidence of postpartum hemorrhage, galactostasis and postpartum depression in the observation group was lower, and the success rate of feeding on the 1st, 3rd and 7th day after delivery was higher in the observation group seven days after intervention. Bilateral early breast sucking within 2 h after delivery is helpful to increase lactation and reduce breast distending pain and postpartum lochia. It also promotes maternal nutritional status and coagulation function, reduces the incidence of postpartum complications, and improves the breastfeeding rate.


Utilization of condensed distillers solubles as nutrient supplement for production of nisin and lactic acid from whey.
The major challenge associated with the rapid growth of the ethanol industry is the usage of the coproducts, i.e., condensed distillers solubles (CDS) and distillers dried grains, which are currently sold as animal feed supplements. As the growth of the livestock industries remains flat, alternative usage of these coproducts is urgently needed. CDS is obtained after the removal of ethanol by distillation from the yeast fermentation of a grain or a grain mixture by condensing the thin stillage fraction to semisolid. In this work, CDS was first characterized and yeast biomass was proven to be the major component of CDS. CDS contained 7.50% crude protein but with only 42% of that protein being water soluble. Then, CDS was applied as a nutrient supplement for simultaneous production of nisin and lactic acid by Lactococcus lactis subsp. lactis (ATCC 11454). Although CDS was able to support bacteria growth and nisin production, a strong inhibition was observed when CDS was overdosed. This may be caused by the existence of the major ethanol fermentation byproducts, especially lactate and acetate, in CDS. In the final step, the CDS based medium composition for nisin and lactic acid production was optimized using response surface methodology.


Significance of collateral circulation in patients with left main coronary artery disease.
Fifty patients with left main coronary artery disease were studied to evaluate the functional role of collateral circulation. The left main was narrowed 50-70% in 22 patients (group I), and more than 70% in 28 patients (group II). Significant disease in the other vessels was equally common in each group. There was no significant difference in the incidence of inter- and intracoronary collaterals in the two groups. Fifteen patients with no collaterals were compared with 35 patients with collaterals, and to a subset of 11 patients with very rich right-to-left collaterals, and there was no significant difference in historic or ECG evidence of old infarction, duration of angina, incidence of unstable angina, left ventricular end-diastolic pressure, cardiac index, ejection fraction, or segmental contraction abnormalities. We conclude that there is no evidence of protective effect of collateral vessels in patients with left main disease.


Visual information gain and the regulation of constant force levels.
Visual information is essential in human motor control, and especially in the continuous modulation of isometric force. The gain of visual feedback, that is, the amount of space used to represent change in force, has been shown to affect both the magnitude and time-dependent properties of variability in the force output. However, little is known regarding the interacting effects of visual gain and target force level on force variability and whether the effects of force level can be mediated by a gain that is adjusted to force level. We examined the effect of different types and levels of visual feedback gain and target force level (1, 2, 4, 8, and 12 N) on the magnitude (standard deviation, SD) and regularity (approximate entropy, ApEn) of isometric force variability. Young adults performed an isometric force task with high and low levels of constant (same gain level for all forces) and scaled (proportional to force level) gain. The magnitude of force variability increased exponentially as a function of force level once the SD was corrected for the limits of the display area. The time-dependent properties of force variability remained constant across force levels when gain was adjusted to force level. These findings suggest that the time-dependent properties of force variability are the result an interaction between visual feedback and task force level demands, while the increases in SD over force levels are primarily due to the invariant properties of human muscle and the motor system.


Enzymatic synthesis of polyglutamate derivatives of 7-hydroxymethotrexate.
7-Hydroxymethotrexate, an important metabolite of methotrexate, is a substrate for folylpolyglutamate synthetase (FPGS) isolated from rat liver and several human leukemia cell lines. The substrate activity it displays over a wide range of concentrations (0-200 microM) is nearly equivalent to that of methotrexate. The 7-hydroxy derivative of dichloromethotrexate is also a substrate for FPGS. The pattern of polyglutamate products synthesized by rat liver FPGS was nearly identical with both 7-hydroxymethotrexate and methotrexate. In addition, conversion of MTX polyglutamates to the corresponding 7-hydroxy compounds was demonstrated using partially purified rabbit liver aldehyde oxidase. The rate of conversion was concentration dependent, and the relative rate decreased as the MTX polyglutamate chain length increased. We propose that 7-hydroxymethotrexate polyglutamates may be formed by initial hydroxylation of methotrexate and subsequent polyglutamate formation or by direct hydroxylation of methotrexate polyglutamates. It was further shown that the relative substrate activity of folate analogs for folylpolyglutamate synthetase is dependent on the source of the enzyme.


Growth hormone-producing pituitary adenoma with giant secretory granules.
Unusually large, oval and pleomorphic secretory granules were noted by electron microscopy in an acidophilic adenoma of the pituitary. The tumor, which was removed by surgery from a 42-year-old woman with elevated blood growth hormone levels and the clinical features of acromegaly, was found to contain growth hormone by the immunoperoxidase technique. This ultrastructural abnormality of secretory granules was not reported so far and was not seen among the 58 cases of growth hormone-producing adenomas investigated in our laboratory. The present case clearly shows that the cytogenesis and cellular composition of pituitary adenomas cannot be determined by solely examining the size and shape of secretory granules.


The relationship between NICU stress and neurodevelopmental outcomes of preterm infants: A multi-center prospective cohort study in China.
To examine the relationship between NICU stress exposure and the neurodevelopmental outcomes of preterm infants. A multicenter, prospective cohort study was conducted between May 2021 and June 2022. Preterm infant participants (28-34 weeks gestational age) were recruited at birth from three NICUs of three tertiary hospitals by convenience sampling. The NICU stress includes acute NICU stress and chronic NICU stress which were measured over the total NICU hospitalization for each infant using the Neonatal Infant Stressor Scale (NISS). Neurodevelopmental outcomes of preterm infants were assessed at 3 months corrected age (CA) using the Ages and Stages Questionnaire, Third Edition (ASQ-3). Of one hundred and thirty preterm infant participants, 108 preterm infants were included into analysis. Results showed that acute NICU stress exposure significantly predicted the neurodevelopmental abnormalities in communication function (RR: 1.001, 95%CI: 1.000-1.001, p = .011), while chronic NICU stress exposure was significantly associated with the problem-solving function (RR: 1.003, 95%CI: 1.001-1.005, p = .002) at 3 months CA. No significant associations were found between NICU stress exposure and other dimensions of neurodevelopmental outcomes, including gross motor, fine motor, and personal-social functions. NICU stress exposure demonstrated a significant predicting relationship with abnormalities in communication and problem-solving functions of preterm infants at 3 months CA. During the NICU hospitalization, neonatal health caregivers should systematically monitor the NICU stress exposure to prevent neurodevelopmental problems in preterm infants.


Diabetic Ketoacidosis Causing Transient Homonymous Hemianopia and Generalized Seizure: A Case Report and Literature Review.
Neurologic complications of hyperglycemia are common. Cases of seizures and hemianopia related to nonketotic hyperglycemia have been reported but are rare with diabetic ketoacidosis. We present clinical, laboratory, and radiologic findings in a patient with diabetic ketoacidosis associated with generalized seizure and homonymous hemianopia, with a literature review of reported cases. Neurologic complications of hyperglycemia are many, but seizure with hemianopia is most commonly associated with nonketotic hyperosmolar hyperglycemia rather than diabetic ketoacidosis. Generalized seizure and retrochiasmal visual field defect are known neurological complications of diabetic ketoacidosis. Like nonketotic hyperosmolar hyperglycemia, these neurological symptoms are transient, and the structural changes in magnetic resonance imaging are usually reversible.


Rat bioassays for methionine availability in 16 food sources.
Methionine availabilities of 16 test proteins were assessed by comparing ten day rat growth response to the test diets and reference (casein) diets. In a preliminary study, various concentrations of methionine and cystine were fed to determine methionine requirements and effect of excess cystine. Results indicated a methionine requirement of about 550 mg per 100 g diet. Cystine had a sparing affect of 50-55%, i.e., about 300 mg could be used to meet methionine requirements. Further additions of cystine (up to 2.6 times methionine) did not affect rat growth. Methionine availabilities were excellent (88-100%) for 15 of the 16 test foods; only pinto beans (58%) were low, but prior evidence indicates that the poor growth response was due to some factor other than availability.


Influence of different assignment conditions on the determination of symmetric homodimeric structures with ARIA.
The ambiguous restraint for iterative assignment (ARIA) approach for NMR structure calculation is evaluated for symmetric homodimeric proteins by assessing the effect of several data analysis and assignment methods on the structure quality. In particular, we study the effects of network anchoring and spin-diffusion correction. The spin-diffusion correction improves the protein structure quality systematically, whereas network anchoring enhances the assignment efficiency by speeding up the convergence and coping with highly ambiguous data. For some homodimeric folds, network anchoring has been proved essential for unraveling both chain and proton assignment ambiguities.


Outcomes identified and prioritised by consumers of Partners in Recovery: a consumer-led study.
Recovery oriented service provisions means focusing on outcomes that are important to consumers themselves rather than to clinicians or services. Partners in Recovery (PIR) is an Australia-wide initiative designed to provide service coordination and brokerage for individuals with severe and persistent mental illness. One PIR service engaged a consumer-led research team to evaluate the service from the perspective of consumers. This consumer-led study was established to explore PIR consumers' perceptions of outcomes they achieved through their involvement with PIR. Data were collected through semi-structured interviews exploring participants' views about and experiences with PIR. Data analysis occurred simultaneously with data collection using constant comparative analysis. Twenty consumers participated. They reported experiencing valued outcomes in six domains: feeling supported; feeling more hopeful and positive about the future; improved mental clarity, focus and order in life; getting out of the house and engaging in positive activity; having a better social life; and improved physical health. Exploring outcomes achieved by PIR consumers, from their own perspective provides a nuanced understanding of the contribution these programs can have in supporting individuals' recovery. Findings from this study highlight the kinds of outcomes consumers achieve when engaged with service coordination and brokerage services. Findings also suggest that outcome measures used in these types of services should focus on recovery outcomes as well as met and unmet needs.


Change in bowel habits during menstruation: are IBD patients different?
Many female inflammatory bowel disease (IBD) patients report symptoms exacerbation before and during menses. Our aim was to characterize their symptoms and specific risk factors in comparison with healthy controls. Female IBD patients aged 18-50 years were asked to fill out a questionnaire recording their demographic and disease characteristics, menstruation history and symptoms. Disease activity was defined by Harvey-Bradshaw index (HBI) for Crohn's disease (CD) patients and modified mayo score for ulcerative colitis (UC) patients. Healthcare providers answering an internet survey assessing bowel symptoms during menstruation served as healthy controls. A total of 139 IBD patients, of whom 100 were CD patients, filled the questionnaire. The mean age was 30.4 [±7.7 standard deviation (SD)], mean disease duration was 7.8 (±6 SD), mean HBI was 4.7 (±3.8 SD), and mean Mayo score was 2.1 (±2.5 SD). A change in bowel habits during menstruation was reported by 72% of CD patients compared with 56% of UC patients (p = 0.07). Out of 258 healthy controls, 93% reported a change in bowel habits during menstruation compared with 68% of IBD patients (p < 0.001). However, other abdominal and constitutional symptoms were significantly more prevalent among IBD patients compared with healthy controls (p < 0.01 for most parameters). Smoking status, biologic treatment, and previous abdominal operation were found to significantly aggravate symptoms during menses in IBD patients. IBD patients experience various symptoms during menses significantly more commonly than healthy women. Smoking, biologic treatment, and previous abdominal operations are risk factors for higher symptomatic burden. Following future validation and research, these results can help in patients' risk stratification and possibly in risk reduction.


Flow diversion treatment for giant intracranial serpentine aneurysms.
Giant serpentine aneurysms (GSAs) are among the most complex and challenging type of intracranial aneurysms. Surgical clipping, bypass, or endovascular parent artery occlusion has been the main treatment of GSAs in the past. However, studies on flow diversion (FD) are limited. Therefore, we reported our experience with patients with GSAs treated with FD. Patients with GSAs treated with FD from 2012 to 2020 in our single center were retrospectively reviewed. Angiographic outcomes were graded according to the O'Kelly-Marotta scale as complete occlusion (D), trace filling (C), entry remnant (B), or aneurysm filling (A). Clinical outcomes were assessed using the modified Rankin scale (mRS) score. We also collected the patients' treatment details and perioperative complications. Thirteen patients with 14 aneurysms were included, including three in the anterior circulation and 11 in the posterior circulation. Grades B-D were found in 72.7% (8/11) of the GSAs. Good prognosis (mRS score, 0-2) was found in 66.7% (8/12) and 50.0% (6/12) of the patients at the 6-month and latest follow-up, respectively. Parent artery occlusion was found in three cases of GSAs. Five postoperative complications were observed, including two minor complications and three major complications. Although reconstructive treatment with FD could be considered as one of the treatment strategies for patients with both anterior and posterior circulation GSAs, however, the risk of complications and parent artery occlusion should be considered.


Binding kinetics of sulfatide with influenza A virus hemagglutinin.
Association of a sulfated galactosyl ceramide, sulfatide, with the viral envelope glycoprotein hemagglutinin (HA) delivered to the cell surface is required for influenza A virus (IAV) replication through efficient translocation of the newly synthesized viral nucleoprotein from the nucleus to the cytoplasm. To determine whether the ectodomain of HA can bind to sulfatide, a secreted-type HA (sHA), in which the transmembrane region and cytoplasmic tail were deleted, was generated by using a baculovirus expression system. The receptor binding ability and antigenic structure of sHA were evaluated by a hemagglutination assay, solid-phase binding assay and hemagglutination inhibition assay. sHA showed subtype-specific antigenicity and binding ability to both sulfatide and gangliosides. Kinetics of sHA binding to sulfatide and GD1a was demonstrated by quartz crystal microbalance (QCM) analysis. QCM analysis showed that the sHA bound with the association rate constant (k on) of 1.41 × 10(4) M(-1) sec(-1), dissociation rate constant (k off) of 2.03 × 10(-4) sec(-1) and K d of 1.44 × 10(-8) M to sulfatide immobilized on a sensor chip. The k off values of sHA were similar for sulfatide and GD1a, whereas the k on value of sHA binding to sulfatide was 2.56-times lower than that of sHA binding to GD1a. The results indicate that sulfatide directly binds to the ectodomain of HA with high affinity.


Metabolomics study of the effect of Danggui Buxue Tang on rats with chronic fatigue syndrome.
Danggui Buxue Tang (DBT), a traditional Chinese medicine (TCM) formula for 'invigorating qi and enriching blood', has been reported to produce a good effect on chronic fatigue syndrome (CFS). However, the related mechanism remains largely undetermined. This study devised a metabolomics approach with GC-MS combined with pattern recognition to estimate the extent to which DBT alleviated CFS induced by food restriction and force swimming in rats. After 4 weeks of treatment, the endurance capability of rats was significantly better, and the motionless time was significantly shorter in the DBT group than in the CFS model group. Moreover, the activities of superoxide dismutase and glutathione peroxidase increased, whereas the levels of malondialdehyde, interleukin 6, and tumor necrosis factor alpha decreased in the DBT treatment group. Fifteen significantly changed metabolites were observed in the serum of rats with CFS, which was reversed markedly by DBT treatment. Metabolic pathway analysis showed that DBT could possibly alleviate CFS in rats by regulating the biosynthesis of phenylalanine, tyrosine, and tryptophan and the metabolism of glycine, serine, threonine, glycerolipid, glyoxylate, dicarboxylate, and tyrosine. It was observed that the metabolism of glycine, serine, and threonine was most closely related to the improvement in CFS by DBT treatment. This study showed that DBT could improve CFS effectively, and metabolomics was a powerful means to gain insights into the TCM formulas against CFS.


A multi-analytical approach to study the chemical composition of total suspended particulate matter (TSP) to assess the impact on urban monumental heritage in Florence.
In the present paper total suspended particulate matter (TSP) was collected at the S. Marco Museum in Florence during seasonal monitoring campaigns carried out in 2013 and 2014. The analyses focused on the determination of TSP chemical composition: main ions, organic carbon (OC), elemental carbon (EC), water soluble organic compounds (WSOC) and levoglucosa which is considered the specific marker for wood burning. The analysis of TSP composition in Florence historical centre is of interest to assess what the damage caused to the surfaces of the urban monumental heritage could be. TSP higher value has been registered during December 2013 (58,6 μg/m3 ± 7,3 μg/m3) while the lower value has been registered in July 2013 (28,5 μg/m3 ± 2,2 μg/m3). OC and EC were also higher in winter with respect to summer. Sulphate is characterized by quite constant values during all the examined periods while ammonium and nitrate were higher in winter. The seasonal contribution of different sources such as combustion processes (from traffic and wood burning) and soil dust resuspension has been evidenced. In particular during winter a high concentration of levoglucosan, the specific marker for wood combustion, has been determined. Finally, all the data obtained were correlated with those previously determined in another study carried out in 2003 at the Baptistery of San Giovanni in Florence in order to evaluate any possible change in the atmospheric pollution composition.


Tartrazine-induced chromosomal aberrations in mammalian cells.
Tartrazine (FD & C Yellow No.5) has been shown to induce chromosomal aberrations in fibroblast cells of Muntiacus muntjac in vitro. M. muntjac cells were exposed to various concentrations of tartrazine (in the 5-20 micrograms/ml range) and were evaluated for induced chromosomal aberrations after two different periods of culture. Total percentages of chromosomal aberrations were significantly increased above control levels in all experimental groups. The results suggest that further studies are needed to determine the potential mutagenic effects of tartrazine.


[Congenital malformations of the atlas: classification and clinical significance].
A classification of congenital anomalies of atlas is proposed in order to make a systematic approach to its radiological diagnosis. Atlas is divided in four areas (posterior arch, union of posterior arch and articulation, articulation and anterior arch) for isolated malformations and two more for occipito-atloid and atlo-axoid anomalies. The common anomalies and its possible clinical relevance in each area are described.


Enhanced framework for COVID-19 prediction with computed tomography scan images using dense convolutional neural network and novel loss function.
Recent studies have shown that computed tomography (CT) scan images can characterize COVID-19 disease in patients. Several deep learning (DL) methods have been proposed for diagnosis in the literature, including convolutional neural networks (CNN). But, with inefficient patient classification models, the number of 'False Negatives' can put lives at risk. The primary objective is to improve the model so that it does not reveal 'Covid' as 'Non-Covid'. This study uses Dense-CNN to categorize patients efficiently. A novel loss function based on cross-entropy has also been used to improve the CNN algorithm's convergence. The proposed model is built and tested on a recently published large dataset. Extensive study and comparison with well-known models reveal the effectiveness of the proposed method over known methods. The proposed model achieved a prediction accuracy of 93.78%, while false-negative is only 6.5%. This approach's significant advantage is accelerating the diagnosis and treatment of COVID-19.


Genome-wide investigation of the GRAS transcription factor family in foxtail millet (Setaria italica L.).
GRAS transcription factors perform indispensable functions in various biological processes, such as plant growth, fruit development, and biotic and abiotic stress responses. The development of whole-genome sequencing has allowed the GRAS gene family to be identified and characterized in many species. However, thorough in-depth identification or systematic analysis of GRAS family genes in foxtail millet has not been conducted. In this study, 57 GRAS genes of foxtail millet (SiGRASs) were identified and renamed according to the chromosomal distribution of the SiGRAS genes. Based on the number of conserved domains and gene structure, the SiGRAS genes were divided into 13 subfamilies via phylogenetic tree analysis. The GRAS genes were unevenly distributed on nine chromosomes, and members of the same subfamily had similar gene structures and motif compositions. Genetic structure analysis showed that most SiGRAS genes lacked introns. Some SiGRAS genes were derived from gene duplication events, and segmental duplications may have contributed more to GRAS gene family expansion than tandem duplications. Quantitative polymerase chain reaction showed significant differences in the expression of SiGRAS genes in different tissues and stages of fruits development, which indicated the complexity of the physiological functions of SiGRAS. In addition, exogenous paclobutrazol treatment significantly altered the transcription levels of DELLA subfamily members, downregulated the gibberellin content, and decreased the plant height of foxtail millet, while it increased the fruit weight. In addition, SiGRAS13 and SiGRAS25 may have the potential for genetic improvement and functional gene research in foxtail millet. Collectively, this study will be helpful for further analysing the biological function of SiGRAS. Our results may contribute to improving the genetic breeding of foxtail millet.


Fat Grafting for Facial Contouring Using Mechanically Stromal Vascular Fraction-Enriched Lipotransfer.
Autologous fat graft has limitations, especially long-term unpredictability of volume maintenance. The mechanical enrichment of fat graft with adipose-derived stem cells (ADSCs) could guarantee the survival of fat grafts. After decantation, washing, and centrifugation of lipoaspirate, the authors carried out histochemical analysis and flow cytometry to determine the best layers for preparing ADSC-enriched fat. After centrifugation, the stromal vascular fraction (SVF) was separated by mechanical dissociation and mixed with another layer of intact adipocytes, which was injected into patients. All patients showed volumetric improvement after a single lipotransfer section, without overcorrection. The method is safe, has low cost, and is easily reproducible.


Automated Quantitative Computed Tomography Versus Visual Computed Tomography Scoring in Idiopathic Pulmonary Fibrosis: Validation Against Pulmonary Function.
The aim of the study was to determine whether a novel computed tomography (CT) postprocessing software technique (CALIPER) is superior to visual CT scoring as judged by functional correlations in idiopathic pulmonary fibrosis (IPF). A total of 283 consecutive patients with IPF had CT parenchymal patterns evaluated quantitatively with CALIPER and by visual scoring. These 2 techniques were evaluated against: forced expiratory volume in 1 second (FEV1), forced vital capacity (FVC), diffusing capacity for carbon monoxide (DLco), carbon monoxide transfer coefficient (Kco), and a composite physiological index (CPI), with regard to extent of interstitial lung disease (ILD), extent of emphysema, and pulmonary vascular abnormalities. CALIPER-derived estimates of ILD extent demonstrated stronger univariate correlations than visual scores for most pulmonary function tests (PFTs): (FEV1: CALIPER R=0.29, visual R=0.18; FVC: CALIPER R=0.41, visual R=0.27; DLco: CALIPER R=0.31, visual R=0.35; CPI: CALIPER R=0.48, visual R=0.44). Correlations between CT measures of emphysema extent and PFTs were weak and did not differ significantly between CALIPER and visual scoring. Intriguingly, the pulmonary vessel volume provided similar correlations to total ILD extent scored by CALIPER for FVC, DLco, and CPI (FVC: R=0.45; DLco: R=0.34; CPI: R=0.53). CALIPER was superior to visual scoring as validated by functional correlations with PFTs. The pulmonary vessel volume, a novel CALIPER CT parameter with no visual scoring equivalent, has the potential to be a CT feature in the assessment of patients with IPF and requires further exploration.


Genome-Wide Meta-Analysis Identifies Three Novel Susceptibility Loci and Reveals Ethnic Heterogeneity of Genetic Susceptibility for IgA Nephropathy.
Eighteen known susceptibility loci for IgAN account for only a small proportion of IgAN risk. Genome-wide meta-analysis was performed in 2628 patients and 11,563 controls of Chinese ancestry, and a replication analysis was conducted in 6879 patients and 9019 controls of Chinese descent and 1039 patients and 1289 controls of European ancestry. The data were used to assess the association of susceptibility loci with clinical phenotypes for IgAN, and to investigate genetic heterogeneity of IgAN susceptibility between the two populations. Imputation-based analysis of the MHC/HLA region extended the scrutiny. Identification of three novel loci (rs6427389 on 1q23.1 [P=8.18×10-9, OR=1.132], rs6942325 on 6p25.3 [P=1.62×10-11, OR=1.165], and rs2240335 on 1p36.13 [P=5.10×10-9, OR=1.114]), implicates FCRL3, DUSP22.IRF4, and PADI4 as susceptibility genes for IgAN. Rs2240335 is associated with the expression level of PADI4, and rs6427389 is in high linkage disequilibrium with rs11264799, which showed a strong expression quantitative trail loci effect on FCRL3. Of the 24 confirmed risk SNPs, six showed significant heterogeneity of genetic effects and DEFA showed clear evidence of allelic heterogeneity between the populations. Imputation-based analysis of the MHC region revealed significant associations at three HLA polymorphisms (HLA allele DPB1*02, AA_DRB1_140_32657458_T, and AA_DQA1_34_32717152) and two SNPs (rs9275464 and rs2295119). A meta-analysis of GWAS data revealed three novel genetic risk loci for IgAN, and three HLA polymorphisms and two SNPs within the MHC region, and demonstrated the genetic heterogeneity of seven loci out of 24 confirmed risk SNPs. These variants may explain susceptibility differences between Chinese and European populations.


Synthesis and Anti-HCV Activity of Sugar-Modified Guanosine Analogues: Discovery of AL-611 as an HCV NS5B Polymerase Inhibitor for the Treatment of Chronic Hepatitis C.
Chronic hepatitis C (CHC) is a major liver disease caused by the hepatitis C virus. The current standard of care for CHC can achieve cure rates above 95%; however, the drugs in current use are administered for a period of 8-16 weeks. A combination of safe and effective drugs with a shorter treatment period is highly desirable. We report synthesis and biological evaluation of a series of 2',3'- and 2',4'-substituted guanosine nucleotide analogues. Their triphosphates exhibited potent inhibition of the HCV NS5B polymerase with IC50 as low as 0.13 μM. In the HCV replicon assay, the phosphoramidate prodrugs of these analogues demonstrated excellent activity with EC50 values as low as 5 nM. A lead compound AL-611 showed high levels of the nucleoside 5'-triphosphate in vitro in primary human hepatocytes and in vivo in dog liver following oral administration.


Bandgap tailoring and enhancing the aromatization in cysteine-based carbon dots.
Cysteine, as a non-aromatic precursor, was used to produce Nitrogen (N) and Sulfur (S) sources for preparing N, S-doped carbon dots (CDs) with tunable luminescence emission. Despite the tremendous investigations, the photoluminescence (PL) mechanism of CDs is still unclear due to its complex core-shell structure, variety of surface functional groups, and structure dependency. This study focuses on controlling aromatization and graphitization processes during the hydrothermal synthesis on CDs by using Citric Acid (CA) and Ammonium persulfate. Detailed characterizations by FTIR spectroscopy, XPS, and HR-TEM are provided to suggest both chemical and bandgap structures. Results reveal that the red-shift of PL occurred due to the graphitization and increasing content of graphitic nitrogen in the core, as well as the Pyridinic and Amine groups creating sub-bands on the surface. These findings resolve the controversy on the PL mechanism of Cysteine-based CDs and provide a general guide for increasing the aromatization and graphitization degree from non-aromatic precursors which clarify the mechanism exploration and structural analysis of other types of CDs.


Cytoplasmic DNA: sources, sensing, and role in aging and disease.
Endogenous cytoplasmic DNA (cytoDNA) species are emerging as key mediators of inflammation in diverse physiological and pathological contexts. Although the role of endogenous cytoDNA in innate immune activation is well established, the cytoDNA species themselves are often poorly characterized and difficult to distinguish, and their mechanisms of formation, scope of function and contribution to disease are incompletely understood. Here, we summarize current knowledge in this rapidly progressing field with emphases on similarities and differences between distinct cytoDNAs, their underlying molecular mechanisms of formation and function, interactions between cytoDNA pathways, and therapeutic opportunities in the treatment of age-associated diseases.


Associations of Organizational Justice with Tinnitus and the Mediating Role of Depressive Symptoms and Burnout-Findings from a Cross-Sectional Study.
Tinnitus refers to the perception of a sound while an external source is absent. Research has identified work-related stress and its potential mental health-related sequelaes, i.e., depression and burnout, as risk factors for tinnitus. Perceived unfairness at work (organizational injustice), which is considered a psychosocial occupational stressor, has been shown to predict depression and burnout but its potential associations with tinnitus remains unaddressed. The aim was to determine the relationship of organizational injustice with tinnitus, and to examine depression and burnout as potential mediators. Cross-sectional data from a sample of 1632 employees were used. Tinnitus was assessed by self-report (n = 207; 13.9 %). Organizational justice and its subcomponents (interactional and procedural justice), burnout, and depressive symptoms were measured by validated questionnaires. Associations were assessed by logistic regressions, and mediation was assessed by maximum likelihood logistic regression estimations. Overall organizational justice, interactional and procedural justice were inversely related to tinnitus (z-score for overall justice: OR = 0.754; 95 % CI = 0.649 to 0.876). These associations were independent of demographics, socioeconomic status, job characteristics (including potential noise exposure), and health behaviors. Mediation analyses suggested a potential mediation by burnout (95 % CI indirect effect -0.188 to -0.066) and depressive symptoms (95 % CI indirect effect -0.160 to -0.043). Parallel multiple mediation analysis revealed that mediation through burnout was significantly larger than through depressive symptoms. Organizational justice appeared inversely related to tinnitus and this association was explained by individual differences in burnout symptoms, suggesting mediation. Longitudinal studies may further help to strengthen the evidence base for prevention of tinnitus through promotion of organizational justice and prevention of burnout.


Antagonistic effects of intravenous or epidural atipamezole on xylazine-induced dorsolumbar epidural analgesia in cattle.
This study was performed to clarify the antagonistic actions of intravenous or epidural atipamezole on the sedative and analgesic effects of xylazine administered between the epidural fat and dura mater through the first interlumbar space in cattle. Cattle received 5 mL of a solution containing 0.05 mg x kg(-1) xylazine in 0.9% saline. Thirty minutes later, 5 mL of 0.9% saline was administered through the same needle (treatment 1) (XSE). In treatments 2 (XAE) and 3 (XAV), 5 mL of a solution containing 0.025 mg x kg(-1) atipamezole in 0.9% saline was administered epidurally or intravenously, respectively. Sedation and analgesia were similar in all three treatment groups and could be reversed by atipamezole given by either route. In the XAV treatment, the flank area relapsed into analgesia 25+/-5.8 min following reversal of the analgesic effect, and was maintained for 112.5+/-63.8 min. The present study confirmed that the sedative and analgesic effects of xylazine are completely reversed by atipamezole and can be influenced by the epidural fat in cattle. Furthermore, it seems probable that analgesia following epidural administration of xylazine is mediated by alpha(2)-adrenergic receptors, not by a local anaesthetic effect.


CRISPR/Cas-mediated knockdown of vacuolar invertase gene expression lowers the cold-induced sweetening in potatoes.
VInv gene editing in potato using CRISPR/Cas9 resulted in knockdown of expression and a lower VInv enzymatic activity resulting in a decrease in post-harvest cold-storage sugars formation and sweetening in potatoes. CRISPR-Cas9-mediated knockdown of vacuolar invertase (VInv) gene was carried out using two sgRNAs in local cultivar of potato plants. The transformation efficiency of potatoes was found to be 11.7%. The primary transformants were screened through PCR, Sanger sequencing, digital PCR, and ELISA. The overall editing efficacy was determined to be 25.6% as per TIDE analysis. The amplicon sequencing data showed maximum indel frequency for potato plant T12 (14.3%) resulting in 6.2% gene knockout and 6% frame shift. While for plant B4, the maximum indel frequency of 2.0% was found which resulted in 4.4% knockout and 4% frameshift as analyzed by Geneious. The qRT-PCR data revealed that mRNA expression of VInv gene was reduced 90-99-fold in edited potato plants when compared to the non-edited control potato plant. Following cold storage, chips analysis of potatoes proved B4 and T12 as best lines. Reducing sugars' analysis by titration method determined fivefold reduction in percentage of reducing sugars in tubers of B4 transgenic lines as compared to the control. Physiologically genome-edited potatoes behaved like their conventional counterpart. This is first successful report of knockdown of potato VInv gene in Pakistan that addressed cold-induced sweetening resulting in minimum accumulation of reducing sugars in genome edited tubers.


A home education program for older adults with hearing impairment and their significant others: a randomized trial evaluating short- and long-term effects.
This paper addresses the development and effectiveness of a home education program. The program, designed for hearing-impaired elders and their significant others (SO), deals with communication strategies and speech reading. Participants were randomly assigned to a training group (hearing aid fitting+home education program) or a control group (hearing aid fitting). The training group included 24 hearing-impaired subjects and 24 SO's. Controls were 24 affected individuals and 22 SO's. Questionnaires addressing emotional response, communication strategies and the IOI-HA, IOI-AI and IOI-SO were used. A repeated measures analysis of variance was applied to test group differences between pre, post, and 6-months follow-up measures. Increased awareness of benefits of speech reading and improved interaction with the SO were observed in the training group only (p < 0.05). No group difference on 'emotional response' was found. IOI-AI and IOI-SO demonstrated favorable attitudes towards the program. Follow-up measures showed improved quality of life and satisfaction in the training group, while a decrease was observed among the controls (p < 0.05). Some effects differed between first-time and experienced hearing aid users. Addition of services to amplification and involvement of the SO are relevant in aural rehabilitation.


Barriers in access to healthcare services for individuals with disorders of sex differentiation in Bangladesh: an analysis of regional representative cross-sectional data.
Worldwide people in disorder of sex development (DSD) faces multiple barriers while seeking their social rights, particularly healthcare services. We aimed to explore the healthcare opportunities available to them, using patterns of healthcare utilization and difficulties faced by DSD population in accessing healthcare services in Bangladesh. Data from a total of 945 DSD population and 71 medical staff were analyzed, collected from three major divisions (Dhaka, Chittagong, and Rajshahi) in Bangladesh during the period of January to December of 2017. A structured questionnaire was used to collect data via face-to-face interviews. Descriptive statistic was used to determine the frequencies of the visit by the DSD population in healthcare facilities as well as to analyze difficulties experienced by the DSD population in getting healthcare services. Multivariate regression analysis was used to explore the association between perceived barriers in getting healthcare services and failures of the DSD population to receive the healthcare services. Present data revealed that around 80% of DSD population sought healthcare services from government healthcare facilities, where the overall success rate in getting healthcare services was less than 50%. The DSD population reported a number of reasons for failures in getting healthcare services, including non-friendly interaction by non-clinical hospital's staff, non-friendly interaction by physicians, public fright as general people do not want to mingle with a DSD person, undesirable excess public interest in DSD individuals, and limitation of the treatment opportunities of hospitals to merely male or female patients. Among the stated reasons, the most frequently reported reason was non-friendly interaction by physicians (50.27%), followed by undesirable excess public interest in DSD individuals (50.16%). DSD population in Bangladesh have limited access to healthcare facilities and facing multiple barriers to get healthcare services. Initiatives from the government and social organizations are important to ensure their access to healthcare services.


Intraoperative Parathyroid Hormone Monitoring: Optimal Utilization.
Intraoperative parathyroid hormone (IOPTH) monitoring is a highly accurate surgical adjunct used to determine the extent of surgery in the setting of primary hyperparathyroidism. It is the successful interpretation of changes in PTH levels that is essential for using this technique in a way to optimize cure. Thus, it is imperative that the surgeon has an understanding of PTH dynamics and carefully chooses the appropriate IOPTH protocol and interpretation criteria that will best predict operative success, minimize unnecessary bilateral exploration, decrease the likelihood of resecting parathyroid glands that are not hypersecreting, and prevent recurrence.


L-Arginine sulfate reduces irreversible protein binding in immobilized metal affinity chromatography.
Immobilized metal affinity chromatography (IMAC) is a powerful technique for capture and purification of relevant biopharmaceuticals in complex biological matrices. However, protein recovery can be drastically compromised due to surface induced spreading and unfolding of the analyte, leading to fouling of the stationary phase. Here, we report on the kinetics of irreversible adsorption of a protease on an IMAC resin in a time span ranging from minutes to several hours. This trend correlated with the thermal data measured by nano differential scanning calorimetry, and showed a time-dependent change in protein unfolding temperature. Our results highlight that 'soft' proteins show a strong time dependent increase in irreversible adsorption. Furthermore, commonly used co-solvents for preservation of the native protein conformation are tested for their ability to reduce fouling. Thermal data suggests that the amino acid l-arginine is beneficial in preventing unfolding, which was confirmed in batch adsorption experiments. The choice of counter-ions has to be considered when using this amino acid. These results show that l-arginine sulfate decelerates the irreversible adsorption kinetics of proteins on the IMAC stationary phase to a greater extent than l-arginine chloride.


Chemical composition and particle size influence the toxicity of nanoscale plastic debris and their co-occurring benzo(α)pyrene in the model aquatic organisms Daphnia magna and Danio rerio.
Little is known about how particle chemical composition and size might influence the toxicity of nanoscale plastic debris (NPD) and their co-occurring chemicals. Herein, we investigate the toxicity of 3 × 1010 particles/L polyethylene (PE, 50 nm), polypropylene (PP, 50 nm), polystyrene (PS, 200 and 600 nm), and polyvinyl chloride (PVC, 200 nm) NPD and their co-occurring benzo(a)pyrene (BaP) to Daphnia magna and Danio rerio. During the 21 days of exposure to PE 50 nm and PS 200 nm, the number of broods produced by D. magna decreased compared to other treatments. Exposure to BaP alone did not produce any effects on the reproduction of the daphnids, however, the mixture of BaP with PS (200 or 600 nm) or with PE (50 nm) reduced the number of broods. Exposure of D. rerio embryos to PE 50 nm, PS 200 nm, and PS 600 nm led to a delay in the hatching. The presence of PS 200 nm and PVC 200 nm eliminated the effects of BaP on the hatching rate of zebrafish. Our findings suggest that data generated for the toxicity of one type of NPD, e.g. PVC or PS may not be extrapolated to other types of NPD.


Origin of congenital cholesteatoma from a normally occurring epidermoid rest in the developing middle ear.
The sites of involvement of congenital cholesteatoma, a lesion which has recently become more frequently recognized, are reviewed from literature sources. There is a propensity for its occurrence, especially when small, in the anterior superior part of the middle ear. The same situation is the precise location of an epidermoid cell rest, the epidermoid formation (EF). This is seen in most fetal ears at the junction of the Eustachian tube with the middle ear near the anterior limb of the tympanic ring, until 33 weeks gestation, when it disappears. Its origin is traced to early fetal life from the ectoderm of the first branchial groove. In embryonic and early in fetal life it seems to act as an organizer in the development of the tympanic membrane and middle ear. It is likely that congenital cholesteatoma is derived from the EF by its continued growth instead of regression. Congenital cholesteatomas show a thinner and flatter matrix than acquired cholesteatomas, probably because the former are most frequently 'closed' and therefore subject to pressure effects from the keratin within the cyst. 'Open' forms also occur in smaller numbers. It is possible that a screening program for congenital cholesteatoma in infants might reduce the incidence of the severe, extended form of the disease.


Luminal hypotonicity increases duodenal mucosal permeability by a mechanism involving 5-hydroxytryptamine.
To investigate whether 5-hydroxytryptamine (5-HT) participates in the mediation of the hypotonicity-induced increase in duodenal mucosal permeability. Proximal duodenum in anaesthetized rats was perfused in situ with a hypotonic NaCl solution and effects on duodenal motility, net fluid flux, mucosal permeability [blood-to-lumen clearance of (51)Cr-ethylenediaminetetraacetic acid (EDTA)] and the release of 5-HT into the luminal solution studied in the presence of the cyclooxygenase inhibitor indomethacin. Perfusion of the duodenum with 50 mm NaCl increased mucosal permeability eightfold, increased the luminal output of 5-HT twofold and induced net fluid absorption. This rise in permeability was enhanced 25% by 5-HT (3 x 10(-3) m), reduced by the 5-HT(3)-receptor antagonists granisetron (10(-4)-3 x 10(-4) m) or ondansetron (10(-5)-10(-4) m) or by the 5-HT(4) receptor antagonist SB 203186 (10(-4) m). The 5-HT(3/4) receptor antagonist tropisetron, at 10(-4) m, did not affect while 3 x 10(-4) and 3 x 10(-3) m augmented the hypotonicity-induced increase in mucosal permeability. Lidocaine (1.1 x 10(-3) m) similarly potentiated while tetrodotoxin (TTX) (5 x 10(-5) m) inhibited the hypotonicity-induced increase in mucosal permeability. Compared with animals treated with indomethacin alone ondansetron and granisetron augmented (by 30-40%) while tropisetron and lidocaine reduced (by 60-70%) the hypotonicity-induced net fluid absorption. Tetrodotoxin and all 5-HT receptor antagonists, except tropisetron, depressed duodenal motility. Luminal hypotonicity increases duodenal mucosal permeability by a neural mechanism involving 5-HT acting on 5-HT(3) and 5-HT(4) receptors. 5-HT also appears to participate in the regulation of the hypotonicity-induced fluid flux.


HabiSign: a novel approach for comparison of metagenomes and rapid identification of habitat-specific sequences.
One of the primary goals of comparative metagenomic projects is to study the differences in the microbial communities residing in diverse environments. Besides providing valuable insights into the inherent structure of the microbial populations, these studies have potential applications in several important areas of medical research like disease diagnostics, detection of pathogenic contamination and identification of hitherto unknown pathogens. Here we present a novel and rapid, alignment-free method called HabiSign, which utilizes patterns of tetra-nucleotide usage in microbial genomes to bring out the differences in the composition of both diverse and related microbial communities. Validation results show that the metagenomic signatures obtained using the HabiSign method are able to accurately cluster metagenomes at biome, phenotypic and species levels, as compared to an average tetranucleotide frequency based approach and the recently published dinucleotide relative abundance based approach. More importantly, the method is able to identify subsets of sequences that are specific to a particular habitat. Apart from this, being alignment-free, the method can rapidly compare and group multiple metagenomic data sets in a short span of time. The proposed method is expected to have immense applicability in diverse areas of metagenomic research ranging from disease diagnostics and pathogen detection to bio-prospecting. A web-server for the HabiSign algorithm is available at http://metagenomics.atc.tcs.com/HabiSign/.


Congenital myopathies.
Congenital myopathies are a heterogeneous group of inherited muscle disorders, characterized by the predominance of particular histopathological features on muscle biopsy, such as cores (central core disease) or rods (nemaline myopathy). Clinically, early onset of the disease, stable or slowly progressive muscle weakness, hypotonia and delayed motor development are common in most forms. As a result, the diagnosis of a subtype of congenital myopathy is largely based on the presence of specific structural abnormalities in the skeletal muscle detected by enzyme-histochemistry and electron microscopy studies. During the last decades there have been significant advances in the identification of the genetic basis of most congenital myopathies. However, there is significant genetic heterogeneity within the main groups of congenital myopathies, and mutations in one particular gene may also cause diverse clinical and morphological phenotypes. Thus, the nosography and nosology in this field is still evolving.


Comparing Central Pain Processing in Individuals With Non-Traumatic Neck Pain and Healthy Individuals: A Systematic Review and Meta-Analysis.
This systematic review and meta-analysis examined the evidence for altered central pain processing in people with nontraumatic neck pain and the relationship among central pain processing, demographics, and pain-related characteristics. Case-control studies reporting measures of altered central pain processing using quantitative sensory testing were reviewed. Standardized mean differences (SMDs) and 95% confidence intervals between people with nontraumatic neck pain and controls were calculated. Meta-analysis was performed using random-effects models when appropriate. Associations between SMDs with demographics and pain-related characteristics were explored on a study level using metaregression. Twenty-six studies were eligible with 25 included for meta-analysis. Meta-analysis demonstrated mechanical hyperalgesia at remote nonpainful sites in the full sample (sample size [n] = 1305, SMD = -0.68) and in the subgroup with moderate/severe disability (n = 165, SMD = -0.86; moderate-quality evidence). Metaregression indicated that remote mechanical hyperalgesia was negatively associated with age (R2 = 25.4%, P = 0.031). Very low- to low-quality evidence of remote cold and heat hyperalgesia and dysfunctional conditioned pain modulation were identified. This review suggests that altered central pain processing is present in people with nontraumatic neck pain and may be associated with disability levels and age. PERSPECTIVE: This review found moderate-quality evidence of mechanical hyperalgesia at remote nonpainful sites in patients with nontraumatic neck pain compared with controls, indicating altered central pain processing. However, more studies are needed to confirm findings from dynamic quantitative sensory testing.


Downregulation of hepatic lipoprotein assembly in rats by fermented products of Monascus pilosus.
Hypercholesterolemia is a major risk factor for atherosclerosis. The fermented products of Monascus sp. have been known for their antihypercholesterolemic effect; however, the studies mostly have focused on the inhibition of cholesterol biosynthesis in liver. In this study, we examined whether fermented products of Monascus pilosus have regulatory effects on the hepatic lipoprotein assembly. Male Wistar rats were fed 1% cholesterol diet for 2 wk. After hypercholesterolemia was induced, the animals were maintained on this cholesterol diet supplemented with M. pilosus-fermented products grown in regular medium/garlic-containing medium or garlic powder for another 6 wk. The concentration of blood lipids and the expression of proteins involved in lipoprotein assembly and hepatic antioxidation were assayed. Maintenance on a 1% cholesterol diet for 2 wk significantly (P < 0.05) raised animals' blood lipid levels and increased the expression of intestinal microsomal triacylglycerol transfer protein, hepatic apolipoprotein B-100, and acyl-coenzyme A:cholesterol acyltransferase. Supplementation of M. pilosus-fermented products or garlic powder significantly (P < 0.05) lowered animals' blood lipid levels and inhibited the expression of intestinal microsomal triacylglycerol transfer protein and hepatic apolipoprotein B-100. The 3-hydroxy-3-methylglutaryl-coenzyme A reductase was downregulated by the M. pilosus-fermented product grown in regular medium but not in garlic-containing medium. The expression of antioxidant enzymes was significantly upregulated by the M. pilosus-fermented product grown in garlic-containing medium. Monascus sp.-fermented products exert the hypocholesterolemic effect by mechanisms other than the inhibition of cholesterol biosynthesis.


JUUL's 2019 Removal of Mint-Flavored Pods and Changes to the Retail Environment of Electronic Nicotine Delivery Systems in Florida and the United States.
The increase in youth electronic nicotine delivery system (ENDS) use coincided with JUUL's rapid rise, which prompted investigations and lawsuits aimed at this leading brand. In response, JUUL discontinued sweet flavors in late 2018, followed by mint flavors in November 2019. We assessed ENDS sales and prices at both the state and national level before and after JUUL's removal of mint flavors. Nielsen retail sales data on ENDS products from convenience and food stores in 4-week aggregates were analyzed between January 2019 and January 2020 in Florida and the United States. Standardized units were created. Unit market share and growth rates were calculated for top brands and flavors in the periods before and after JUUL's mint removal. Average prices within brand and product type were calculated. Following JUUL's removal of mint in November 2019, JUUL's market share dropped from over 66% in Florida and the United States to 37.1% (Florida) and 55.1% (United States). In January 2020, the second leading brands were Puff Bar (15.0%) in Florida and Vuse (18.1%) in the United States. Mint market share decreased and share of all other flavor categories increased, particularly menthol and concept. Total ENDS sales increased in Florida but decreased in the United States. Average prices of ENDS devices decreased. While JUUL's actions led to a decline in its sales, Puff Bar emerged and menthol and concept flavors experienced growth. Findings also demonstrate how changes by influential brands differentially affect purchase patterns at the national and state level. These findings support the growing body of evidence that tobacco industry self-regulation, with selective flavor removal by the leading ENDS brand in this case, is insufficient to reduce total ENDS sales, including sales of flavored products which are preferred by youth. Results suggest that brand and flavor substitution compensated for the removal of mint JUUL pods. Understanding changes to the ENDS market in response to industry actions, at both the state and national level, can inform future regulation and interventions. These findings can also inform efforts to prevent and reduce youth ENDS use.


Differentiation of genes extracted from non-viable versus viable micro-organisms in environmental samples using ethidium monoazide bromide.
Differentiation of DNA derived from viable or non-viable microorganisms within mixed microbial communities continues to be one of the greatest challenges in molecular studies of environmental samples. A novel method developed for microbial food pathogens is tested here on environmental samples. This technique involves the use of ethidium monoazide bromide (EMA) for the distinction of live/dead cells. In non-viable cells EMA intercalates into the DNA which prevents amplification by PCR. We adapted and evaluated the EMA technique for soil, elemental sulfur and river biofilm samples. Quantitative PCR determined that EMA suppressed 99.99% of E. coli LKI gfp+ signal in non-viable cultures and 100.00% when the cultures were added to soil samples. The same technique was also successful at suppressing DNA amplification from spiked non-viable cells in elemental sulfur samples by 100.00%, but not in three Saskatchewan River biofilms. In sub Antarctic soil, EMA-Q-PCR was used to detect the prevalence of a functional gene, amoA, and this was closely correlated to nitrification activity measurements. The ability of EMA to differentiate between viable and non-viable populations in soil was confirmed by the similarity of the 16S rRNA denaturing-gradient-gel electrophoresis DNA fingerprint of EMA treated soil and the 16S rRNA cDNA fingerprint of non-EMA treated soil. The EMA technique effectively suppressed amplification of non-viable spiked controls, closely mirrored activity assays and yielded community composition profiles similar to rRNA techniques. The use of EMA in soil effectively suppressed amplification of non-viable organism DNA, however it was not effective in biofilm samples and EMA partially inhibited amplification of viable organism DNA in elemental sulfur samples.


Osteoarthritis year in review 2020: epidemiology & therapy.
This personal choice of research themes and highlights from within the past year (1 May 2019 to 14 April 2020) spans descriptive, analytical-observational, and intervention studies. Descriptive estimates of the burden of osteoarthritis continue to underscore its position as a leading cause of disability worldwide, but whose burden is often felt greatest among disadvantaged and marginalised communities. Many of the major drivers of that burden are known but epidemiological studies continue the important work of elaborating on their timing, dose, specificity, and reversibility and placing them within an appropriate multi-level framework. A similar process of elaboration is seen also in studies (re-)estimating the relative benefits and risks of existing interventions, in some cases helping to identify low-value care, unwarranted variation, and initiating processes of deprescribing and decommissioning. Such research need not engender therapeutic nihilism. Our review closes by highlighting some emerging evidence on the efficacy and safety of novel therapeutic interventions and with a selective roll-call of methodological and meta-research in OA illustrating the continued commitment to improving research quality.


The effects of an induced adenosine deaminase deficiency on T-cell differentiation in the rat.
Inherited deficiency of the enzyme adenosine deaminase (ADA) has been found in a significant proportion of patients with severe combined immunodeficiency disease and inherited defect generally characterized by a deficiency of both B and T cells. Two questions are central to understanding the pathophysiology of this disease: (1) at what stage or stages in lymphocyte development are the effects of the enzyme deficiency manifested; (2) what are the biochemical mechanisms responsible for the selective pathogenicity of the lymphoid system. We have examined the stage or stages of rat T-cell development in vivo which are affected by an induced adenosine deaminase deficiency using the ADA inhibitors, erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) and 2'-deoxycoformycin (DCF). In normal rats given daily administration of an ADA inhibitor, cortical thymocytes were markedly depleted; peripheral lymphocytes and pluripotent hemopoietic stem cells (CFU-S) all were relatively unaffected. Since a deficiency of ADA affects lymphocyte development, the regeneration of cortical and medullary thymocytes and their precursors after sublethal irradiation was used as a model of lymphoid development. By Day 5 after irradiation the thymus was reduced to 0.10-0.5% of its normal size; whereas at Days 9 and 14 the thymus was 20-40% and 60-80% regenerated, respectively. When irradiated rats were given daily parenteral injections of the ADA inhibitor plus adenosine or deoxyadenosine, thymus regeneration at Days 9 and 14 was markedly inhibited, whereas the regeneration of thymocyte precursors was essentially unaffected. Thymus regeneration was at least 40-fold lower than in rats given adenosine or deoxyadenosine alone. Virtually identical results were obtained with both ADA inhibitors, EHNA and DCF. The majority of thymocytes present at Day 9 and at Day 14 in inhibitor-treated rats had the characteristics of subcapsular cortical thymocytes which are probably the most ancestral of the thymocytes. Thus, an induced ADA deficiency blocked the proliferation and differentiation of subcapsular cortical thymocytes which are the precursors of cortical and medullary thymocytes.


Comparison of Effects of Statin Use on Mortality in Patients With Heart Failure and Preserved Versus Reduced Left Ventricular Ejection Fraction.
Randomized trials showed no survival benefit with statin therapy in heart failure (HF) patients with reduced ejection fraction (HFrEF). Whether these results are generalizable to HF patients with preserved ejection fraction (HFpEF) or with mid-range ejection fraction is unclear. In a cohort of 13,440 patients with HF, 9,903 (73.7%) were treated with statins. The association between statin use and all-cause mortality was assessed with Cox proportional hazard regression models and survival time inverse probability weighting propensity scores analyses. Multivariable Poisson regression models with robust error variance were applied to estimate the rate ratios (RR) for hospitalization. The association between statin treatment and clinical outcomes differed by ejection fraction group. In patients with HFpEF, statin use was associated with reduced mortality (hazard ratio [HR] 0.73, 95% confidence interval [CI] 0.66 to 0.81, p <0.001; average treatment effect [ATE] 0.48, 95% CI 0.13 to 0.84, p = 0.007) and reduced all-cause hospitalization (RR 0.82, 95% CI 0.76 to 0.89, p <0.001). In contrast, in patients with HFrEF, no significant association was observed between statin use and mortality (HR 0.86, 95% CI 0.74 to 1.0, p = 0.054; ATE 0.41, 95% CI -0.09 to 0.93, p = 0.11) or hospitalization (RR 0.92, 95% CI 0.82 to 1.04, p = 0.17). Similarly, in patients with mid-range ejection fraction, there was no significant association with reduced mortality (HR 0.76, 95% CI 0.60 to 0.95, p = 0.02, ATE 0.3, 95% CI -0.84 to 1.43, p = 0.61) or hospitalization (RR 1.07, 95% CI 0.9 to 1.27, p = 0.44). In conclusion, statin use was associated with improved clinical outcomes in patients with HFpEF but not in patients with HFrEF or mid-range ejection fraction.


Isoelectric focusing studies in Brazilian Indians--uncovering variation of ORM, AHSG and IF.
Allele frequencies for the orosomucoid 1 (ORM1), orosomucoid 2 (ORM2), alpha-2-HS-glycoprotein (AHSG) and complement component I (IF) loci were studied in 393 individuals of three Brazilian Indian tribes. In the ORM1 locus only two alleles were observed among the Urubu-Kaapor, while five were found among the Pacaás Novos. The frequency of ORM1*1 was similar in these two tribes (0.734 and 0.715, respectively) but departed more markedly among the Parakanã (0.870). Variation for ORM2 locus was found among the Pacaás Novos only, with ORM2*3 being observed in just three individuals. A new variant (AHSG*PN) was found in the AHSG system. Frequency for AHSG*1 was unexpectedly low in the three tribes, especially, among the Pacaás Novos, where the prevalence (0.145) is the lowest considering other data reported thus far. For IF locus, variability was also restricted to only one trible (Urubu-Kaapor) and attributed to a new polymorphic allele, IF* A3.


Acute biliary pancreatitis. The roles of laparoscopic cholecystectomy and endoscopic retrograde cholangiopancreatography.
Since January 1990, we have treated 113 patients for gallstone pancreatitis; 59 with laparoscopic cholecystectomy (LC), 50 with open cholecystectomy, and 4 with ERCP/sphincterotomy only. In the LC group, 47 had LC during the index admission and 12 underwent delayed LC. Fifty patients had open cholecystectomy, 47 during the index admission. ERCPs were performed in 43 of the 113 patients; CBD stones were identified in 19/43 (44%) and removed endoscopically in 18 (95%). The ERCP complication rate was 6.5%. In total, CBD stones were identified in 29/113 patients (26%). Patients who had imaging of the CBD within the first 4 days from onset of symptoms were more likely to have stones identified than were those patients who were studied after 5 days. Recurrent pancreatitis occurred in in five of 11 patients (45%) who had a > or = 30-day delay to definitive treatment. We conclude that LC can be safely performed in most patients during the index admission for gallstone pancreatitis. This policy should reduce the 30-50% risk of recurrent pancreatitis associated with a delayed operation. ERCP is a helpful adjunct for CBD stones.


Operative mortality following surgery for colorectal cancer.
From 1973 to 1986 at the Baviere Hospital, University of Liège, 476 patients underwent surgery for colorectal cancers. The overall operative mortality rate was 13.4 per cent. The following postoperative life-threatening complications occurred: myocardial infarction (14 per cent of the postoperative deaths) during the first three postoperative days; bronchopneumonia (27 per cent of the postoperative deaths) mainly during the first postoperative week; pulmonary embolism (17 per cent of the postoperative deaths) mainly during the second postoperative week; anastomotic leakage and cerebrovascular accident (14 and 8 per cent of the postoperative deaths respectively) mainly during the third and fourth postoperative weeks. Specific risk factors were chronic obstructive airways disease associated with a higher incidence of postoperative bronchopneumonia, and previous myocardial infarction associated with postoperative myocardial infarction and pulmonary embolism. General risk factors were old age and emergency procedure, both of which were associated with a higher incidence of infectious complications. The operative mortality rate fell from 20.1 per cent in the 1973-79 period to 7.8 per cent in the 1980-86 period.


Temporal assessment of caspase activation in experimental models of focal and global ischemia.
Rodent models of focal and global ischemia were used to examine caspase activation. Several readouts were employed on identical tissue to provide correlative measurement of caspase induction, activation and enzymatic activity. In a rat focal ischemia model, caspase-3 enzymatic activity, as recorded by DEVD-AMC cleavage, peaked in penumbral cortex at 6-12 h following ischemia, correlating with increases in caspase 3-cleaved substrates of PARP and alpha-spectrin and subsequent disappearance of caspase-3 zymogen. Although induction of caspases 8 and 2 proteins was detectable as early as 6 h following ischemia, examination of the same tissues for caspase 8 or 2 enzymatic activities did not show significant modulation up to 12 h after ischemic insult. Caspase 9 induction was evident only after 24 h postischemia and did not correlate with elevated LDHD-AMC cleavage. Following global ischemia in gerbils, levels of caspase-3 enzyme activity peaked at 12 h in hippocampal tissue extracts. Cleaved caspase-3 signal was prominent in NeuN-positive layers in the CA1 region 6-12 h following ischemia. Interestingly, strong caspase-3 immunoreactivity was also detected in the subgranular zone of the dentate gyrus, a known region of ischemia-induced neurogenesis. Caspase-3 activation may be responsible for the loss of these cells, thereby hindering the endogenous recovery process.


Three new species of the pill millipede genus Hyleoglomeris Verhoeff, 1910, from northern Thailand (Diplopoda, Glomerida, Glomeridae).
Seven species of the basically warm temperate to tropical Eurasian genus Hyleoglomeris are currently known from Thailand, including three new ones: H. hongkhraiensis sp. n. and H. aurea sp. n. from Chiang Mai Province, and H. cavicola sp. n. from Sukhothai Province. A new distribution map and a key to all Hyleoglomeris species presently known to occur in Thailand are given.


Naturally occurring CD4+ CD25+ FOXP3+ T-regulatory cells are increased in chronic myeloid leukemia patients not in complete cytogenetic remission and can be immunosuppressive.
Clinical presentation of chronic myeloid leukemia (CML) requires not only the deregulated tyrosine kinase BCR-ABL, but also the failure of an immune response against BCR-ABL-expressing cells. T-cell responses against BCR-ABL and other antigens are well-described, but their relevance to the in vivo control of CML is unclear. The suppressive role of naturally occurring T regulatory (T-reg) cells in antitumor immunity is well-established, although little is known about their role in modulating the T-cell response to BCR-ABL. Naturally occurring T-reg cells were characterized and quantified by flow cytometry in 39 CML patients and 10 healthy donors. Their function was studied by observing their effect on responses to purified protein derivative, a recall antigen, and on the response of an autologous T-cell line recognizing BCR-ABL. T-reg cells were CD4(+), CD25(+), FOXP3(+), CD127(low), and CD62L(high). T-reg numbers in patients in complete cytogenetic remission were significantly lower than in patients not in complete cytogenetic remission (p < 0.01). T-reg cell depletion using anti-CD25 selection enhanced proliferative responses to purified protein derivative. Furthermore, the interferon-γ and/or granzyme-B production of effector cells specific for viral peptides or a BCR-ABL HLA-A3-restricted peptide was inhibited when autologous T-reg cells were present. Taken together, these data suggest a role for T-reg cells in limiting immune responses in CML patients and this may include immune responses to BCR-ABL. The increased frequency of T-reg cells in patients with high levels of BCR-ABL transcripts indicates that an immune mechanism may be important in the control of CML.


Oleic acid stimulates complete oxidation of fatty acids through protein kinase A-dependent activation of SIRT1-PGC1α complex.
Fatty acids are essential components of the dynamic lipid metabolism in cells. Fatty acids can also signal to intracellular pathways to trigger a broad range of cellular responses. Oleic acid is an abundant monounsaturated omega-9 fatty acid that impinges on different biological processes, but the mechanisms of action are not completely understood. Here, we report that oleic acid stimulates the cAMP/protein kinase A pathway and activates the SIRT1-PGC1α transcriptional complex to modulate rates of fatty acid oxidation. In skeletal muscle cells, oleic acid treatment increased intracellular levels of cyclic adenosine monophosphate (cAMP) that turned on protein kinase A activity. This resulted in SIRT1 phosphorylation at Ser-434 and elevation of its catalytic deacetylase activity. A direct SIRT1 substrate is the transcriptional coactivator peroxisome proliferator-activated receptor γ coactivator 1-α (PGC1α), which became deacetylated and hyperactive after oleic acid treatment. Importantly, oleic acid, but not other long chain fatty acids such as palmitate, increased the expression of genes linked to fatty acid oxidation pathway in a SIRT1-PGC1α-dependent mechanism. As a result, oleic acid potently accelerated rates of complete fatty acid oxidation in skeletal muscle cells. These results illustrate how a single long chain fatty acid specifically controls lipid oxidation through a signaling/transcriptional pathway. Pharmacological manipulation of this lipid signaling pathway might provide therapeutic possibilities to treat metabolic diseases associated with lipid dysregulation.


The complete mitochondrial genome of Pseudochauhanea macrorchis (Monogenea: Chauhaneidae) revealed a highly repetitive region and a gene rearrangement hot spot in Polyopisthocotylea.
The complete mitochondrial genome of Pseudochauhanea macrorchis was determined and compared with other monogenean mitochondrial genomes from GenBank. The circular genome was 15,031 bp in length and encoded 36 genes (12 protein-coding genes, two ribosomal RNAs, and 22 transfer RNAs) typically found in flatworms. Structures of the mitochondrial genome were mostly concordant with that known for Microcotyle sebastis and Polylabris halichoeres, but also contained two noted features-a gene rearrangement hot spot and the highly repetitive region (HRR) in major non-coding region (NCR). The gene rearrangement hot spot located between the cox3 and nad5 genes, including a cluster of tRNA genes, nad6 gene and one major NCR. The HRR seemed to be a unique feature of the polyopisthocotylean mitochondrial genomes. In conclusion, the present study provided new molecular data for future studies of the comparative mitochondrial genomics and also served as a resource of markers for the studies of species populations and monogenean phylogenetics.


The Quality and Content of Internet-Based Information on Orthopaedic Sports Medicine Requires Improvement: A Systematic Review.
To evaluate the quality and content of internet-based information available for some of the most common orthopaedic sports medicine terms. A search of the PubMed, Embase, and Cochrane databases following PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-analyses) guidelines was performed. All English-language literature published from 2010 to 2020 discussing information quality pertaining to orthopaedic sports medicine terms was included. Outcomes included the search engines used, number and type of websites evaluated, platform, and quality scoring metrics. Descriptive statistics are presented. This review includes 21 studies. Of these, 3 evaluated both the upper and lower extremity. Twelve focused on either the upper or lower extremity, most commonly rotator cuff tears (3 of 12) and/or anterior cruciate ligament pathologies (7 of 12). The most common engines were Google (18 of 21), Bing (16 of 21), Yahoo (16 of 21), YouTube (3 of 21), Ask (3 of 21), and AOL (2 of 21). The average number of media files assessed per study was 87 ± 55. Website quality was assessed with DISCERN (7 of 21), Flesch-Kincaid (9 of 21), Health on the Net (7 of 21), and/or Journal of the American Medical Association Benchmark (7 of 21) scores. YouTube was evaluated with Journal of the American Medical Association Benchmark scores (1.74 ± 1.00). Image quality was reported in 2 studies and varied with search terminology. The results of this systematic review suggest that physicians should improve the quality of online information and encourage patients to access credible sources when conducting their own research. Doctors can and should play an active role in closing the gap between the level of health literacy of their patients and that of most common online resources.


Modifying interpretation biases: Effects on symptomatology, behavior, and physiological reactivity in social anxiety.
The present study investigated the effects of computerized interpretation training and cognitive restructuring on symptomatology, behavior, and physiological reactivity in an analogue social anxiety sample. Seventy-two participants with elevated social anxiety scores were randomized to one session of computerized interpretation training (n = 24), cognitive restructuring (n = 24), or an active placebo control condition (n = 24). Participants completed self-report questionnaires focused on interpretation biases and social anxiety symptomatology at pre and posttraining and a speech task at posttraining during which subjective, behavioral, and physiological measures of anxiety were assessed. Only participants in the interpretation training condition endorsed significantly more positive than negative interpretations of ambiguous social situations at posttraining. There was no evidence of generalizability of interpretation training effects to self-report measures of interpretation biases and symptomatology or the anxiety response during the posttraining speech task. Participants in the cognitive restructuring condition were rated as having higher quality speeches and showing fewer signs of anxiety during the posttraining speech task compared to participants in the interpretation training condition. The present study did not include baseline measures of speech performance or computer assessed interpretation biases. The results of the present study bring into question the generalizability of computerized interpretation training as well as the effectiveness of a single session of cognitive restructuring in modifying the full anxiety response. Clinical and theoretical implications are discussed.


Optogenetic ion pumps differ with respect to the secondary pattern of K+ redistribution.
We recently reported that strong activation of the optogenetic chloride pump, halorhodopsin leads to a secondary redistribution of K+ ions into the cell, through tonically open, "leak" K+ channels. Here we show that this effect is not unique to halorhodopsin but is also seen with activation of another electrogenic ion pump, archaerhodopsin. The two opsins differ however in the size of the rebound rise in extracellular potassium, [K+ ]o , after the end of activation, which is far larger with halorhodopsin than for archaerhodopsin activation. Multiple linear regression modeling indicates that the variance in the postillumination surge in [K+ ]o was explained both by the size of the preceding, illumination-induced drop in [K+ ]o and also by the type of opsin. These data provide additional support for the hypothesis that intense chloride-loading of cells, as occurs naturally following intense bursts of GABAergic synaptic bombardment, or artificially following halorhodopsin activation, is followed by extrusion of both Cl- and K+ coupled together. We discuss this with respect to the pattern of [K+ ]o rise that occurs at the onset of seizure-like events.


Safety of Atrovent® CFC-free inhaler: respiratory events reported from an observational cohort study in England.
The aim of the study was to identify any unexpected clinical events associated with starting the new CFC-free formulation of Atrovent® MDI in general practice in England. An active surveillance cohort study was conducted with a focus on selected clinical events, including respiratory symptoms, in past users of Atrovent® CFC MDI ('switchers') and Atrovent® naïve users. Incidence density rate ratios (with 99% confidence intervals) for events occurring in the first 3 months of exposure (risk period-ID1-3 ) compared to 3 months prior to starting treatment (reference period-IDR ) were calculated. The cohort consisted of 13 211 patients (median age 70 years, 50.1% female; 63.5% prior users of Atrovent® CFC MDI ('switchers')). Common respiratory events occurred at higher rates after starting treatment than before for switchers, for example lower respiratory tract infection (LRTI) [ID1 /IDR = 1.45 (99% CI: 1.17, 1.81)] and worsening asthma [ID1 /IDR = 1.58 (99% CI: 1.00, 2.51)]. Of these events only LRTI was significant for Atrovent® naïve patients [ID1 /IDR = 1.42 (99% CI: 1.04, 1.95)]. The results of this study suggest effect modification of risk as a result of prior Atrovent® CFC MDI use. Overall, Atrovent® CFC-free MDI appeared to be reasonably well tolerated in the immediate postmarketing period and the safety profile appeared similar to that of the CFC formulation.


[Generators of pathologically enhanced excitation as determinant structures in the spinal cord].
Using tetanus toxin as a tool to disturb the inhibition, enhanced excitation generators were created in the anterior horns of lumbar segments of the left and right sides of albino rat spinal cord. The generators worked at different regimens: the "left-side" generator originating under conditions of a more prolonged toxin action, on being activated by trigger stimulation, produced first tonic and then intermittent activity or periodic spontaneous discharges; the "right-side" generator produced only tonic activity. After the inhibition of one of the generators by glycine the other continued working in its regimen. Activation of one of the generators was followed by inhibition of the effect of the other. In case of separate activation of one of the generators all pools of the spinal and supraspinal motor neurons reproduced the activity pattern of the generator working at that moment. Thus, the latter played the role of the hyperactive structure determining the system behaviour, i.e. the determinant. The evidence obtained is analyzed from the point of view of the general concept on the role of the determinant structures in the nervous system activity and the theory of generator mechanisms of the neuropathological syndromes characterized by the system hyperactivity.


Modeling tumor-host interactions of chronic lymphocytic leukemia in xenografted mice to study tumor biology and evaluate targeted therapy.
Chronic lymphocytic leukemia (CLL) cells depend on microenvironmental factors for proliferation and survival. In particular, the B-cell receptor (BCR) and nuclear factor- κB (NF-κB) pathways are activated in the lymph node (LN) microenvironment. Thus, model systems mimicking tumor-host interactions are important tools to study CLL biology and pathogenesis. We investigated whether the recently established NOD/scid/γc(null) (NSG) mouse xenograft model can recapitulate the effects of the human microenvironment. We assessed, therefore, tumor characteristics previously defined in LN-resident CLL cells, including proliferation, and activation of the BCR and NF-κB pathways. We found that the murine spleen (SP) microenvironment supported CLL cell proliferation and activation to a similar degree than the human LN, including induction of BCR and NF-κB signaling in the xenografted cells. Next, we used this model to study ibrutinib, a Bruton's tyrosine kinase inhibitor in clinical development. Ibrutinib inhibited BCR and NF-κB signaling induced by the microenvironment, decreased proliferation, induced apoptosis and reduced the tumor burden in vivo. Thus, our data demonstrate that the SP of xenografted NSG mice can, in part, recapitulate the role of the human LN for CLL cells. In addition, we show that ibrutinib effectively disrupts tumor-host interactions essential for CLL cell proliferation and survival in vivo.


Preparation of amino-modified cellulose aerogels and adsorption on typical diclofenac sodium contaminant.
A new functional cellulose aerogel (Cell@PEI) with high adsorption efficiency was prepared for the removal of diclofenac sodium (DCF) by ammonification cross-linked polyethyleneimine (PEI) with the surface of cellulose. The fabricated Cell@PEI adsorbent was characterized using field emission scanning electron microscopy (FESEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), etc. The results demonstrated that the Cell@PEI exhibited a distinct three-dimensional cell structure and was rich in functional groups, i.e., -OH, C = O, -NH2, and C = C. The Cell@PEI presented a stable crystal structure and large specific surface area (241.41 m2·g-1), which was approximately 42 times as much as bare cellulose aerogel (5.82 m2·g-1). In addition, a series of adsorption experiments showed that the adsorbent had good adsorption performance for DCF with a maximum adsorption capacity of 294.12 mg·g-1. Furthermore, the adsorption of DCF on Cell@PEI was well corresponded with the Langmuir isotherm and pseudo-second-order adsorption model. Thermodynamic study proved that adsorption was a spontaneous exothermic reaction. Moreover, the introduction of PEI into Cell@PEI aerogel enhanced the electrostatic interaction and hydrogen bonding, promoting DCF adsorption. Importantly, the Cell@PEI aerogel could be reused up to five times desorbed by NaOH (0.5 mol/L). Considering the above results, the fabricated aerogel material can be applied to remove organic pollutants.


Potential growth and control of Escherichia coli O157:H7 in soft hispanic type cheese.
This study examined growth and control of two enterohemorrhagic Escherichia coli serotype O157:H7 strains in a soft Hispanic type cheese (Queso Fresco). Cheese was made in the laboratory using a commercial procedure and after inoculation it was stored under vacuum at temperatures ranging from 8 to 30 degrees C. The minimum temperature that allowed growth of E. coli in the unmodified normal cheese (pH 6.6) was 10 degrees C. No growth of either strains was observed during a 2 month storage period at 8 degrees C. Accumulated data from the growth studies were used for development of models relating square root of 1/LT (LT = lag time) and specific growth rate, to temperature. The effect of selected antimicrobials (potassium sorbate, sodium benzoate, and sodium lactate) added to milk or cheese on the growth and survival of these pathogens at various storage temperatures was also evaluated. Addition of sodium benzoate (0.3%) to cheese (pH 6.6) or addition of potassium sorbate (0.3%) to cheese (pH 6) made from milk acidified to pH 5.9 with propionic acid, had a significant impact on delaying or preventing growth of the pathogens.


High-throughput RNAi screen for essential genes and drug synergistic combinations in colorectal cancer.
Metastatic colorectal cancer is a leading cause of cancer death. However, current therapy options are limited to chemotherapy, with the addition of anti-EGFR antibodies for patients with RAS wild-type tumours. Novel drug targets, or drug combinations that induce a synergistic response, would be of great benefit to patients. The identification of genes that are essential for cell survival can be undertaken using functional genomics screens. Furthermore, performing such screens in the presence of a targeted agent would allow the identification of combinations that result in a synthetic lethal interaction. Here, we present a dataset containing the results of a large scale RNAi screen (815 genes) to detect essential genes as well as synergistic combinations with targeted therapeutic agents using a panel of 27 colorectal cancer cell lines. These data identify genes that are essential for colorectal cancer cell survival as well as synthetic lethal treatment combinations using novel computational approaches. Moreover, this dataset could be utilised in combination with genomic profiling to identify predictive biomarkers of response.


Lipid profile in children with insulin dependent diabetes mellitus.
To evaluate serum lipids in children with type 1 diabetes mellitus and its relation to glycemic control, in comparison with matched controls. Basrah Maternity and Children Hospital, Basrah, Iraq. Lipid profile was studied in 32 children (17 males and 15 females) with type 1 diabetes mellitus from October 1999 to September 2000. Seventy seven non-diabetic children (35 males and 42 females) admitted with acute illness in the same hospital were selected as controls. The age and BMI was similar in both groups. The mean total cholesterol, triglycerides, HDL-C, LDL-C, VLDL-C and LDL-C/HDL-C ratio were significantly higher in diabetic children compared to control group. There was no significant difference in the levels of Apo O and Apo B in both groups. Children with type 1 diabetes should be screened for serum lipids as significant lipid abnormalities are related to glycemic control. Both can be improved with dietary management and insulin therapy.