Genomic heterogeneity underlies multidrug resistance in Pseudomonas aeruginosa: A population-level analysis beyond susceptibility testing.
Pseudomonas aeruginosa is a persistent and difficult-to-treat pathogen in many patients, especially those with Cystic Fibrosis (CF). Herein, we describe a longitudinal analysis of a series of multidrug resistant (MDR) P. aeruginosa isolates recovered in a 17-month period, from a young female CF patient who underwent double lung transplantation. Our goal was to understand the genetic basis of the observed resistance phenotypes, establish the genomic population diversity, and define the nature of sequence evolution over time. Twenty-two sequential P. aeruginosa isolates were obtained within a 17-month period, before and after a double-lung transplant. At the end of the study period, antimicrobial susceptibility testing, whole genome sequencing (WGS), phylogenetic analyses and RNAseq were performed in order to understand the genetic basis of the observed resistance phenotypes, establish the genomic population diversity, and define the nature of sequence changes over time. The majority of isolates were resistant to almost all tested antibiotics. A phylogenetic reconstruction revealed 3 major clades representing a genotypically and phenotypically heterogeneous population. The pattern of mutation accumulation and variation of gene expression suggested that a group of closely related strains was present in the patient prior to transplantation and continued to change throughout the course of treatment. A trend toward accumulation of mutations over time was observed. Different mutations in the DNA mismatch repair gene mutL consistent with a hypermutator phenotype were observed in two clades. RNAseq performed on 12 representative isolates revealed substantial differences in the expression of genes associated with antibiotic resistance and virulence traits. The overwhelming current practice in the clinical laboratories setting relies on obtaining a pure culture and reporting the antibiogram from a few isolated colonies to inform therapy decisions. Our analyses revealed significant underlying genomic heterogeneity and unpredictable evolutionary patterns that were independent of prior antibiotic treatment, highlighting the need for comprehensive sampling and population-level analysis when gathering microbiological data in the context of CF P. aeruginosa chronic infection. Our findings challenge the applicability of antimicrobial stewardship programs based on single-isolate resistance profiles for the selection of antibiotic regimens in chronic infections such as CF.


Dietary influence on the insulin function in the epididymal fat cell of the Wistar rat. I. Effect of type of fat.
For at least 3-4 weeks, young male Wistar rats were fed semisynthetic diets containing sunflowerseed oil, palm oil, olive oil, linseed oil, cocoa butter and coconut oil as dietary fat. The type of dietary fat had little effect on body weight, epididymal fat pad weight and on the diameter of fat cells isolated from the epididymal fat pads, but the rats fed the linseed oil-containing diet had a lower epididymal fat pad weight. The fatty acid composition of the triacylglycerol fractions of the fat pads correlated well with those of the dietary fats. The correlations with the fatty acid composition of the phospholipids of the fat pads were less pronounced. High responses to insulin in the epididymal fat cells were obtained with sunflowerseed oil, linseed oil and olive oil, whereas low responses were found for cocoa butter, palm oil or coconut oil. Rather than the amount of polyunsaturated fatty acids or the ratio of polyunsaturated to saturated fatty acids, the amount of saturated fatty acids with 12, 14 or 16 carbon atoms appeared to be the most important parameter in determining the maximal insulin response. A negative correlation was found between the amount of saturated fatty acids in the diet and the extent of insulin response. The modulating effects of the dietary type of fat on the response to insulin cannot be fully explained by changes in the number of insulin receptors on the fat cell surface as determined by insulin binding but must, at least partially, be ascribed to postreceptor effects.


Rubella vaccine in postpubertal women. Experience in Western Washington State.
Nearly two thousand nonimmune, postpubertal females given rubella vaccine in western Washington state in 1970 to 1974 were followed up for acute reactions, inadvertent pregnancy onset, and (776 women) seroresponse. Low-level prevaccination immunity appears to explain most of the apparent vaccine failure (11.6%). The 27 vaccine-complicated pregnancies identified resulted in 17 apparently normal infants (nine from nonimmune mothers). The remaining ten were terminated by abortion (four in nonimmune women), but no abortus yielded rubella virus. Frequency of post-vaccination complaints (largely joint-related) varied with age (higher in those over 25 years), with vaccine (higher after HPV-77-DE-5 than after Cendehill strain virus), with stage of menstrual cycle when vaccine was given, and with method of contraception.


Genistein inhibits Na+/Ca2+ exchange activity in primary rat cortical neuron culture.
We have examined the possible regulatory effect of tyrosine kinase activity on Ca2+ transport observed in the cultured rat cortical neurons. Na+/Ca2+ exchange was studied using cells cultured for various time periods. A nearly two fold increase in Ca2+ uptake was seen when comparing 3 day and 9 day cultures. Western blot analysis also showed a two fold increase in Na+/Ca2+ exchanger (NCX1) protein levels as cells matured in culture. To study the effect of genistein (a specific tyrosine kinase inhibitor) cells were incubated with 100 microM genistein (in 1% DMSO) for 1 hour before the assay of Na+/Ca2+ exchange activity. There was a significant decrease of Ca2+ uptake in genistein treated neurons (control: 4.596+/-0.205 nmol/mg protein/15 min, n=12; genistein: 1.420+/-0.131 nmol/mg protein/15 min, n=12, mean+/-S.E. P<0.001). Daidzein, an inactive analog of genistein and phorbol myristate acetate (PMA), a PKC activator were without effect. The results suggest that as cells mature in culture, Na+/Ca2+ exchange capacity increases, as a result of greater protein expression. Exposure to genistein inhibited Ca2+ uptake suggesting that the exchanger may be modulated by tyrosine phosphorylation.


[Examination of late pulmonary toxicity in children treated for malignancies].
The present investigation was based on a survey in 2005, in which the authors found pulmonary function abnormalities in survivors of childhood cancer, who were treated with anticancer therapy. The purpose of the present study was to follow-up childhood cancer survivors and detect late pulmonary toxicity. Lung function test was performed with spirometry in 26 survivors participated in this study (10 females and 16 males; mean age, 19.4 years at the time of the second follow-up evaluation). The average time periods from treatment until the first and second follow-up evaluation were 4.5 and 10 years, respectively. The authors found 14 patients with pathological pulmonary function tests results at the time of the first follow-up evaluation, from which 7 patients had obstructive, 5 patients had mixed and 2 patients had restrictive abnormalities. However, there were only 6 patients who had abnormal pulmonary function at the time of the second follow-up evaluation (2 patients with obstructive and 4 patients with restrictive pulmonary function tests (p<0.05). Restrictive pulmonary disorder was detected in only small part of the treated patients. The obstructive pulmonary abnormalities caused by the treatment showed an improving tendency over time.


Carbon dioxide laser enucleation of polypoid vocal cords.
Polypoid vocal cords have routinely been treated by endoscopic vocal cord stripping, often-times resulting in prolonged hoarseness postoperatively. Submucosal CO2 laser enucleation of the polypoid tissue, with preservation of a mucosal flap on the medial edge of the cord, has proved to be a valuable improvement. The surgical procedure is described and results are presented which suggest that voice quality is better earlier than is the case after vocal cord stripping.


DNA hybridization of cervical tissues.
The increasing frequency of cervical neoplasia among younger women and the increased invasiveness of these tumors has led to a considerable growth in research into this disease. Conventional methods (epidemiology, cytology, and immunology), while being extremely useful, also have significant limitations. Recent advances in techniques for the manipulation of DNA now make it possible to analyze tissues for the presence of viral genomes. This review introduces these techniques and describes their application to the search for herpes simplex virus and human papillomavirus sequences in cervical tissue. The significance of the findings both for the mechanism of transmission of the disease, and also the consequences for early detection and hence more successful treatment, are also discussed.


NK cells eliminate Epstein-Barr virus bound to B cells through a specific antibody-mediated uptake.
Epstein Barr virus (EBV) causes a highly prevalent and lifelong infection contributing to the development of some malignancies. In addition to the key role played by T cells in controlling this pathogen, NK cells mediate cytotoxicity and IFNγ production in response to EBV-infected B cells in lytic cycle, both directly and through antibody (Ab)-dependent activation. We recently described that EBV-specific Ab-dependent NK cell interaction with viral particles (VP) bound to B cells triggered degranulation and TNFα secretion but not B cell lysis nor IFNγ production. In this report we show that NK cell activation under these conditions reduced B cell transformation by EBV. NK cells eliminated VP from the surface of B cells through a specific and active process which required tyrosine kinase activation, actin polymerization and Ca2+, being independent of proteolysis and perforin. VP were displayed at the NK cell surface before being internalized and partially shuttled to early endosomes and lysosomes. VP transfer was encompassed by a trogocytosis process including the EBV receptor CD21, together with CD19 and CD20. Our study reveals a novel facet of the antibody-dependent NK cell mediated response to this viral infection.


Cystatin C as a Potential Blood Biomarker for Sarcoidosis: A Case Report.
Sarcoidosis is a multi-organ medical condition that is characterized by the formation of granulomas. We aimed to identify a correlation between each sarcoidosis blood biomarker and cystatin C (Cys-C) in sarcoidosis patients. We report a case of a 60-year-old man with sarcoidosis. The correlation between his Cys-C and each blood biomarker level and that between each blood biomarker and serum creatinine levels were determined using linear regression. Serum Cys-C correlated with each blood biomarker of sarcoidosis, while creatinine did not. These findings suggest that Cys-C is a potential blood biomarker for sarcoidosis.


Preparatory attention in visual cortex.
Top-down attention is the mechanism that allows us to selectively process goal-relevant aspects of a scene while ignoring irrelevant aspects. A large body of research has characterized the effects of attention on neural activity evoked by a visual stimulus. However, attention also includes a preparatory phase before stimulus onset in which the attended dimension is internally represented. Here, we review neurophysiological, functional magnetic resonance imaging, magnetoencephalography, electroencephalography, and transcranial magnetic stimulation (TMS) studies investigating the neural basis of preparatory attention, both when attention is directed to a location in space and when it is directed to nonspatial stimulus attributes (content-based attention) ranging from low-level features to object categories. Results show that both spatial and content-based attention lead to increased baseline activity in neural populations that selectively code for the attended attribute. TMS studies provide evidence that this preparatory activity is causally related to subsequent attentional selection and behavioral performance. Attention thus acts by preactivating selective neurons in the visual cortex before stimulus onset. This appears to be a general mechanism that can operate on multiple levels of representation. We discuss the functional relevance of this mechanism, its limitations, and its relation to working memory, imagery, and expectation. We conclude by outlining open questions and future directions.


Clinical variants of pityriasis rosea.
Pityriasis rosea (PR) is a common erythemato-squamous dermatosis which almost always, is easily diagnosed. Mostly the disease presents in its classical form. However, clinical dermatology is all about variations and PR is not an exception. Variants of the disease in some cases may be troublesome to diagnose and confuse clinicians. Prompt diagnosis and treatment of the condition becomes necessary to avoid unnecessary investigations. We hereby review and illustrate atypical presentations of the disease, including diverse forms of location and morphology of the lesions, the course of the eruption, and its differential diagnoses.


Suppression of the purgative action of rhein anthrone, the active metabolite of sennosides A and B, by indomethacin in rats.
Rhein anthrone (12.48 mg kg-1) produces watery and mucoid diarrhoea approximately 20 min after intracaecal administration to rats. Pretreatment with the prostaglandin (PG) biosynthesis inhibitor indomethacin (10 mg kg-1, i.p.) only delayed and did not completely block the onset of the induced diarrhoea. Rhein anthrone stimulated PGE2 release into the rat colonic lumen and the increased release was depressed by indomethacin. Rhein anthrone also accelerated large intestinal transit and this acceleration could be partly inhibited by indomethacin, which was probably responsible for the delay in the onset of diarrhoea. Indomethacin prevented the enhanced water, K+ and mucus secretion and the reduced Na+ absorption in the colon which were induced by rhein anthrone. The net water secretion could not be reversed to net absorption and the mucus secretion was only slightly depressed by indomethacin. Thus, our findings suggest that other mechanisms, together with the PG-dependent mechanism, are involved in the purgative action of rhein anthrone in rats.


Pharmacological treatment of ischaemic heart disease. Monotherapy vs combination therapy.
Non-invasive treatment continues to be the mainstay of anti-ischaemic therapy. The imbalance between myocardial oxygen supply and demand provides the basis for myocardial ischaemia, which may present as symptomatic, asymptomatic or, in most cases, a mixture of the two. Both symptomatic and silent ischaemia are major prognostic indicators in patients with coronary artery disease and treatment should therefore be directed towards both the amelioration of symptoms and the resolution of the signs of ischaemia. Anti-ischaemic therapy may decrease oxygen demand, increase myocardial oxygen supply, or both. Interventional therapies, such as percutaneous transluminal coronary angioplasty and bypass grafting, improve supply but do not alter demand. Drug therapy is associated with a variety of effects upon both supply and demand, depending upon the agent used. Nitrates alter both myocardial oxygen supply and demand, while beta-blockers decrease myocardial oxygen demand. Calcium channel blockers reduce after load and myocardial contractility and, thus, lower oxygen demand, while the coronary artery relaxation that occurs in response to their use acts to increase supply. The use of combination therapy is considered by many to be the most rational approach to the treatment of myocardial ischaemia, in that it allows maximal reduction in demand and increase in supply.


Downregulation of HMGA2 by the pan-deacetylase inhibitor panobinostat is dependent on hsa-let-7b expression in liver cancer cell lines.
Inhibitors of protein deacetylases represent a novel therapeutic option for cancer diseases due to their effects on transcriptional regulation by interfering with histones acetylation and on several other cellular pathways. Recently, their ability to modulate several transcription factors and, interestingly, also co-factors, which actively participate in formation and modulation of transcription complexes was shown. We here investigate whether HMGA2 (High Mobility Group AT-2 hook), a nuclear non-histone transcriptional co-factor with known oncogenic properties, can be influenced by the novel pan-deacetylase inhibitor panobinostat (LBH589) in human hepatocellular carcinoma models. Panobinostat strongly downregulated HMGA2 in HepG2 and Hep3B cells; this effect was mediated by transcriptional upregulation and promotion of the maturation of the tumorsuppressor miRNA hsa-let-7b, which could inhibit HMGA2 expression via RNA interference pathways. siRNA knockdown of HMGA2 or transfection of hsa-let-7b mimicking oligonucleotides confirmed the role of HMGA2 in regulating cell proliferation and apoptosis in liver cancer cell lines. Co-incubation with panobinostat showed an additive effect on inhibition of cell proliferation using an impedance-based real-time cell analyzer. Treatment of HepG2 xenografts with panobinostat also led to a downregulation of HMGA2 in vivo. These findings show that pan-deacetylase inhibitors also modulate other signaling pathways and networks than histone modifications to influence cell fate.


Embryonic chicken cornea and cartilage synthesize type IX collagen molecules with different amino-terminal domains.
We have analyzed embryonic chicken cornea for the presence of type IX collagen mRNA and protein. Using RNA transfer blot analysis, we demonstrate that alpha 1(IX) and alpha 2(IX) mRNAs are expressed by corneal epithelial cells at the time that the primary stromal components are synthesized. The levels of the mRNAs decrease with increasing developmental age and are barely detectable at day 11 of development. In contrast, type IX collagen protein is detectable by immunofluorescence at days 5 and 6 and undetectable by day 8. Using probes specific for alpha 1(IX) and alpha 2(IX) mRNAs, we demonstrate that the size of alpha 2(IX) mRNA is the same in cornea as in chondrocytes, the major source of type IX collagen. However, the alpha 1(IX) mRNA is about 700 nucleotides shorter in the cornea than in cartilage because the corneal form of the mRNA does not contain the 5' region that encodes the non-triple-helical amino-terminal globular domain of cartilage type IX collagen. Therefore, corneal type IX collagen must lack this domain. This structural modulation of an extracellular matrix protein is likely to contribute to the functional differences between cartilage matrix and the early corneal stroma, both of which are rich in type II collagen.


Mycobacterium avium-Mycobacterium intracellular complex-induced suppression of T-cell proliferation in vitro by regulation of monocyte accessory cell activity.
Heat-killed whole Mycobacterium avium-Mycobacterium intracellulare complex (MAC) and its lipid component impaired the capacity of human peripheral blood mononuclear cells to proliferate in vitro in response to concanavalin A (ConA), purified protein derivative of tuberculin (PPD), and to a lesser degree, phytohemagglutinin stimulation. Inhibition by MAC was not contingent upon prior exposure of the donor to MAC or other mycobacteria and occurred with lymphocytes from tuberculin-negative as well as -positive subjects. The suppression was not due to the toxicity of MAC. The suppression by MAC was not blocked by indomethacin. Adherent cell depletion and cell mixing experiments with T cells indicated that monocytes and not T cells were a major contributor to the immunosuppression observed. However, neither interleukin-1 production nor the expression of HLA-DR (Ia antigen) by monocytes was suppressed by MAC treatment. On the other hand, treatment of monocytes with MAC or MAC-derived lipid resulted in significant decreases in CD11b, a member of the leukocyte function-associated molecule-1 and LeuM3 (CD14) molecule. Anti-CD18 (beta-chain of the leukocyte function-associated molecule-1 family) monoclonal antibody had suppressive effects on ConA- and PPD- but not phytohemagglutinin-induced in vitro lymphocyte blastogenesis. We suggest that MAC and MAC-derived lipid suppress the ConA- and PPD-induced T-cell proliferations by blocking the expression of accessory molecules on the surfaces of monocytes which might be involved in nonspecific monocyte-T-cell interactions and not by inhibiting either monocyte Ia antigen expression or interleukin-1 production by monocytes.


A Genome-wide Analysis of Human Pluripotent Stem Cell-Derived Endothelial Cells in 2D or 3D Culture.
A defined protocol for efficiently deriving endothelial cells from human pluripotent stem cells was established and vascular morphogenesis was used as a model system to understand how synthetic hydrogels influence global biological function compared with common 2D and 3D culture platforms. RNA sequencing demonstrated that gene expression profiles were similar for endothelial cells and pericytes cocultured in polyethylene glycol (PEG) hydrogels or Matrigel, while monoculture comparisons identified distinct vascular signatures for each cell type. Endothelial cells cultured on tissue-culture polystyrene adopted a proliferative phenotype compared with cells cultured on or encapsulated in PEG hydrogels. The proliferative phenotype correlated to increased FAK-ERK activity, and knockdown or inhibition of ERK signaling reduced proliferation and expression for cell-cycle genes while increasing expression for "3D-like" vasculature development genes. Our results provide insight into the influence of 2D and 3D culture formats on global biological processes that regulate cell function.


Cognitive Decline in Alzheimer's Disease: Limited Clinical Utility for GWAS or Polygenic Risk Scores in a Clinical Trial Setting.
Introduction: Alzheimer's disease (AD) is a progressive and irreversible neurological disease. The genetics and molecular mechanisms underpinning differential cognitive decline in AD are not well understood; the genetics of AD risk have been studied far more assiduously. Materials and Methods: Two phase III clinical trials measuring cognitive decline over 48 weeks using Alzheimer's Disease Assessment Scale-cognitive subscale (ADAS-cog, n = 2060) and Clinical Dementia Rating-Sum of Boxes (CDR-SB, n = 1996) were retrospectively genotyped. A Genome-Wide Association Study (GWAS) was performed to identify and replicate genetic variants associated with cognitive decline. The relationship between polygenic risk score (PRS) and cognitive decline was tested to investigate the predictive power of aggregating many variants of individually small effect. Results: No loci met candidate gene or genome-wide significance. PRS explained a very small percentage of variance in rates of cognitive decline (ADAS-cog: 0.54%). Conclusions: These results suggest that incorporating genetic information in the prediction of cognitive decline in AD currently appears to have limited utility in clinical trials, consistent with small effect sizes estimated elsewhere. If AD progression is more heritable soon after disease onset, genetics may have more clinical utility.


Genome assembly anchored QTL map of bovine chromosome 14.
Bovine chromosome 14 (BTA14) has been widely explored for quantitative trait loci (QTL) and genes related to economically important traits in both dairy and beef cattle. We reviewed more than 40 investigations and anchored 126 QTL to the current genome assembly (Btau 4_0). Using this anchored QTL map, we observed that, in dairy cattle, the region spanning 0 - 10 Mb on BTA14 has the highest density QTL map with a total of 56 QTL, mainly for milk production traits. It is very likely that both somatic cell score (SCS) and clinical mastitis share some common QTL in two regions: 61.48 Mb - 73.84 Mb and 7.86 Mb - 39.55 Mb, respectively. As well, both ovulation rate and twinning rate might share a common QTL region from 34.16 Mb to 65.38 Mb. However, there are no common QTL locations in three pregnancy related phenotypes: non-return rate, pregnancy rate and daughter pregnancy rate. In beef cattle, the majority of QTL are located in a broad region of 15 Mb - 45 Mb on the chromosome. Functional genes, such as CRH, CYP11B1, DGAT1, FABP4 and TG, as potential candidates for some of these QTL, were also reviewed. Therefore, our review provides a standardized QTL map anchored within the current genome assembly, which would enhance the process of selecting positional and physiological candidate genes for many important traits in cattle.


The effect of in vivo rotator cuff muscle contraction on glenohumeral joint translation: An ultrasonographic and electromyographic study.
The proposed stabilizing mechanism of rotator cuff muscles is to limit excessive humeral head translation. However, an accurate measurement of glenohumeral joint translation in vivo has been challenging. We aimed to measure the effect of rotator cuff muscle contraction on glenohumeral joint translation using real time ultrasound (RTUS) and electromyography. Twenty healthy adults with no history of shoulder pathology were recruited. Six intramuscular electrodes were inserted in the rotator cuff muscles (supraspinatus, upper and lower infraspinatus, teres minor, upper and lower subscapularis). Anterior and posterior glenohumeral translations were measured in testing conditions (with and without translation force, with and without isometric internal and external rotation), in two positions (shoulder neutral, abduction) and views (anterior, posterior). There was reduced glenohumeral translation with rotator cuff muscle contraction in the neutral anterior (F2,38=17.8, p<0.01), neutral posterior (F1.6,31.0=44.3, p<0.01) and abducted posterior (F1.5,28.8=5.2, p<0.02) positions. There were also differences between the amount of translation limited by anterior and posterior rotator cuff muscles in response to anterior and posterior translation forces (p<0.05), indicating that their activity was, to a certain extent, direction specific. For example, in both neutral and abducted positions, contraction of the posterior rotator cuff muscles, infraspinatus and teres minor, appeared to tether anterior translation of the humeral head. Our results confirm that the rotator cuff functions as a stabilizer of the glenohumeral joint by limiting humeral head translation and this is likely to be in a direction-specific manner.


Low oxygen treatment prior to cold storage decreases the incidence of bitter pit in 'Golden Reinders' apples.
The effect of subjecting 'Golden Reinders' apples to a low O(2) pre-treatment (LOT; 1-2% O(2)) was evaluated as a strategy to decrease the rate of bitter pit (BP) incidence after standard cold storage (ST). Immediately after harvest, apples were stored for 10 days at 20 degrees C under low O(2). Thereafter, apples were cold-stored (0-4 degrees C) for 4 months and changes were monitored in terms of BP incidence, fruit quality traits and mineral element concentrations. After 4 months cold storage, LOT apples presented a 2.6-fold decrease in the rate of BP incidence (14%) versus the values obtained for standard cold-stored fruits (37% BP incidence). LOT increased flesh firmness, total soluble solids and titratable acidity as compared to the quality traits determined for cold-stored fruits. Lower cortex Ca and Mg concentrations as compared to ST apples were determined in association with LOT, 2 months after cold storage. Application of a LOT prior to cold storage may be a promising strategy to reduce the incidence of BP and preserve fruit quality, which should be further investigated.


[High-performance liquid chromatography-mass spectrometry for determining olmesartan in human plasma].
To establish a simple and rapid HPLC-MS method for determining the contents of olmesartan in human plasma. Plasma were precipitated with trifluoroacetic acid, then analyzed on an HyPurity C(18) column (150 mm 2.1 mm, 5 microm). Samples at 40 degrees celsius;. The mobile phase consisted of water-methanol- acetonitrile(14:60:26) with a flow rate of 0.22 ml/min. The lower limit of qualification was 25 microg/L. The calibration curve was linear over the range of 25-3200 microg/L (r=0.9998), with the intra-day and inter-day RSD less than 15%. The method is sensitive, rapid and suitable for the study of pharmacokinetics and bioavailability of olmesartan.


Correction of complete maxillary crossbite with severe crowding using Hyrax expansion and fixed appliance.
An 18-year-old Thai man who presented with a secondary cleft palate, maxillary hypoplasia and severe crowding was treated by rapid maxillary expansion and fixed orthodontic appliances. Initial assessment found skeletal Class III malrelationship and dental Class II malocclusion with anterior and bilateral posterior crossbites. Camouflage orthodontic treatment was planned using a rapid maxillary expansion appliance and correcting crowding with extraction all four premolar teeth. A Hyrax appliance and vertical loop arch wire were placed for maxillary arch expansion of 9.5 mms at first molars and canines, and 5.5 mms at the premolars and obtained positive overjet. Both acceptable skeletal and soft tissue relationships and satisfactory occlusion were produced. After 14 months of postoperative follow-up, the occlusal result was stable and no skeletal reversals could be detected.


Behaviors that discriminate ADHD in children and adolescents: primary symptoms, symptoms of comorbid conditions, or indicators of functional impairment?
The objectives of this study were to evaluate whether behaviors that differentiate children and adolescents with ADHD from those without are related to the primary diagnostic criteria (i.e., inattention and impulsivity-hyperactivity), symptoms of comorbid conditions, functional impairment, or a combination, and to determine whether behaviors that discriminate are consistent between the key developmental stages of childhood and adolescence. Parents and teachers rated children and adolescents with a formal diagnosis of ADHD and those without any mental, physical, or emotional disorders on the multidimensional Behavior Assessment System for Children (2nd ed., BASC-2). Results indicated that behaviors that discriminate children and adolescents with ADHD from those without were associated with primary symptoms, symptoms of comorbid conditions, and indicators of functional impairment. Teachers rated atypicality as the strongest discriminator of children with ADHD from those without and learning problems as the strongest discriminator of adolescents. Parents rated hyperactivity as the strongest discriminator for children and attention problems for adolescents. Activities of daily living was the third strongest discriminator of adolescents as rated by parents. Findings supported the need for a multidimensional view of ADHD that adds emphasis to issues of functional impairment in routine, daily activities important to success in school and interpersonally. Findings showed that conceptualizing and assessing behaviors demonstrated by children and adolescents with ADHD as a whole instead of only equating a diagnosis of ADHD to hyperactivity, inattention, and impulsivity will establish a foundation for understanding, interpreting, and addressing students' academic and behavioral needs across the home and school.


Modulation of gene expression by selected fatty acids in human breast cancer cells.
The effect of linoleic acid (LNA, n-6) and eicosopentanoic acid (EPA, n-3) was investigated on the parental MCF-7 cells and those transfected with v-H-ras. Transfection of v-H-ras oncogene renders the normally estrogen dependent MCF-7 cells estrogen independent. The effects of LNA and EPA in both the cell lines were measured by their ability to enhance the steady state cytoplasmic mRNA levels of 1-8 and 2-5 A, two genes whose transcription is enhanced in various cells in response to interferons (IFNs). The modulatory effect of these fatty acids on the level of an oncogene HER-2/neu was also investigated. Our results indicate that both the fatty acids induce mRNA 1-8 in parental MCF-7 cells but not in the ras transfected cell line, although the gene is induced in both cell lines in response to IFNs. mRNA 2-5A was not induced by LNA or EPA in either of the cell lines. HER-2/neu levels were enhanced by EPA in MCF-7-ras cells. Our data provide evidence to support the concept that selected, nutritionally relevant, fatty acids can regulate gene expression in vitro. These fatty acids can also induce second messenger signals similar to the ones generated by IFNs.


Availability of body magnesium during magnesium deficiency.
Extensive experimental and clinical studies in animals and humans during the past 25 years have yielded considerable data regarding the availability of internal magnesium (Mg) stores for the maintenance of extracellular and critical tissue concentrations of Mg during periods of Mg deprivation. The bulk of body Mg is present in bone and skeletal muscle, and it is these two sites that provide essentially all the available Mg. In the rat, approximately 15% of bone Mg (equivalent to 1.5 mmol/kg body weight) can be lost, whereas less than one tenth of that amount is available from skeletal muscle. In the human, up to 35% of bone Mg can be lost, but the mean loss in extant studies is 18% (equivalent to 1.2 mmol/kg body weight). In contrast to the rat, up to 40% of human skeletal-muscle Mg can be lost with an average loss of 15% (equivalent to 0.45 mmol/kg). In the human, the bone and skeletal-muscle Mg pools can provide an average of 1.7 mmol/kg body weight equivalent to 15% of total body Mg. Release of Mg from these stores appears to depend on the presence of hypomagnesemia which may also result in small, significant and potentially adverse Mg losses from certain vital organs such as the heart, kidney and brain. The liver and other organs appear not to lose Mg despite Mg deprivation although intracellular Mg shifts of importance cannot be ruled out. These data indicate that the body reserve to combat Mg depletion is not designed to protect the extracellular Mg pool and certain critical organs from Mg deficiency. A continuous optimal intake of Mg is needed for good nutrition and health.


The role of cystatin-C and fetuin-A in the determination of early atherosclerotic risk in psoriasis patients.
Psoriasis is associated with an increased risk of atherosclerosis. The objective of this study is evaluate the relationship between carotid intima-media thickness (CIMT) serum and cystatin-C, fetuin-A levels in determining the increased risk of subclinical atherosclerosis in psoriasis patients. In this study, age and gender compatible 80 psoriasis patients and 78 healthy individuals were included. For both groups, serum cystatin-C, fetuin-A, high-sensitivity C-reactive protein (hs-CRP), oxidized low-density lipoprotein (ox-LDL), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglyceride, total cholesterol, and creatinine levels were recorded and carotid intima-media thickness (CIMT) was measured via B-mode ultrasonography by cardiology department. In binary comparisons between patient and control groups, cystatin-C, fetuin-A, hs-CRP, and CIMT values were higher in psoriasis patients and there was a statistically significant difference (P < .05). In control group, serum HDL-C level was statistically significant higher (P < .05). There was no statistically significant difference in hs-CRP, ox-LDL, LDL-C, triglyceride, total cholesterol, and creatinine levels between the groups (P > .05). Our study supported that psoriasis is a risk factor for development of atherosclerosis and we think that cystatin-C can be used as an important marker in determining subclinical atherosclerosis in patients with psoriasis.


A labile inhibitor blocks immunoglobulin kappa-light-chain-gene transcription in a pre-B leukemic cell line.
The murine pre-B leukemic cell line 70Z/3 contains both an unrearranged immunoglobulin kappa-light-chain gene and a functionally rearranged but silent kappa-light-chain gene. Mitogenic stimulation of growing 70Z/3 cells with bacterial lipopolysaccharide (LPS) activates kappa-light-gene transcription and results in a 10- to 20-fold increase in cytoplasmic kappa-light-chain mRNA. The induction of kappa gene expression by LPS was probed by using an inhibitor of protein synthesis. Concomitant treatment of 70Z/3 cells with LPS and cycloheximide failed to block kappa-light-chain mRNA accumulation, indicating that new protein synthesis is not required for the activation of kappa-gene expression. Treatment of 70Z/3 cells with cycloheximide alone resulted in kappa-mRNA induction equivalent to those produced by LPS alone. The kappa mRNA synthesized in the presence of cycloheximide was intact and able to direct the synthesis of kappa light chains. Nuclear transcription assays revealed that cycloheximide, like LPS, activated kappa-gene transcription. These findings indicate that the trans-acting factors necessary for kappa-light-chain-gene transcription are present in pre-B cells, but their activity is blocked by short-lived inhibitory proteins.


Thrombin regulation of synaptic plasticity: implications for physiology and pathology.
Thrombin, a serine protease involved in the coagulation cascade has been recently shown to affect neuronal function following blood-brain barrier breakdown. Several lines of evidence have shown that thrombin may exist in the brain parenchyma under normal physiological conditions, yet its role in normal brain functions and synaptic transmission has not been established. In an attempt to shed light on the physiological functions of thrombin and Protease Activated Receptor 1 (PAR1) in the brain, we studied the effects of thrombin and a PAR1 agonist on long term potentiation (LTP) in mice hippocampal slices. Surprisingly, different concentrations of thrombin affect LTP through different molecular routes converging on PAR1. High thrombin concentrations induced an NMDA dependent, slow onset LTP, whereas low concentrations of thrombin promoted a VGCCs, mGluR-5 dependent LTP through activated Protein C (aPC). Remarkably, aPC facilitated LTP by activating PAR1 through an Endothelial Protein C Receptor (EPCR)-mediated mechanism which involves intracellular calcium stores. These findings reveal a novel mechanism by which PAR1 may regulate the threshold for synaptic plasticity in the hippocampus and provide additional insights into the role of this receptor in normal and pathological conditions.


Incidence of cancers following orthotopic liver transplantation in a single center: comparison with national cancer incidence rates for England and Wales.
The incidence of de novo cancers is increased in liver allograft recipients but there are few data assessing the extent of the increased risk compared with a matched population. A retrospective study of 1,778 adults transplanted between January 1982 and March 2004, followed for a median of 65 months. The observed cancer incidence was compared with age-, sex-, and calendar year-matched expected cancer rates in England and Wales population. In all, 141 (7.9%) developed a new cancer. There was an increase in the incidence of all tumors compared with that expected (Standardized Incidence Ratio (SIR) 207, 95% CI 174-244, P < 0.001); the greatest increase was seen in lymphoid tumors (SIR 1026, 95% CI 608-1621, P < 0.001), skin cancers (SIR 580, 95% CI 432-763, P < 0.001), and cancer of the large bowel (SIR 496, 95% CI 290-774, P < 0.001). Large bowel cancer was more common in those patients with ulcerative colitis than those without (SIR 2727 vs. 347) and in older patients. Females had a greater risk of lung cancer than males (SIR 336 vs. 56). There is an increased incidence of tumors following liver transplantation. Although the absolute risk of cancer is low, we found that the increase in risk is greater in the younger aged recipients than the older ones. Increased awareness of colon cancer is needed especially in older patients and those with ulcerative colitis. There should be awareness for the high lung cancer incidence in females. Increased surveillance for breast and cervical cancer is not necessary.


Chromosome microarrays in human reproduction.
Chromosome microarray (CMA) testing allows automatic and easy identification of large chromosomal abnormalities detectable by conventional cytogenetics as well as the detection of submicroscopic chromosomal imbalances. A PubMed search was performed in order to review the current use of CMA testing in the field of human reproduction. Articles discussing the use of CMA in the preimplantation setting, ongoing pregnancies, miscarriages and patients with reproductive disorders were considered. A high rate of concordance between conventional methods of detecting chromosomal abnormalities [e.g. fluorescence in situ hybridization (FISH), karyotyping] and CMA was reported in the prenatal setting with CMA providing more comprehensive and detailed results as it investigates the whole genome at higher resolution. In preimplantation genetic screening, CMA is replacing FISH and the selection of embryos based on CMA has already resulted in live births. For ongoing pregnancies and miscarriages, CMA eliminates tissue culture failures and artifacts and allows a quick turnaround time. The detection of submicroscopic imbalances [or copy number variants (CNVs)] is beneficial when the imbalance has a clear clinical consequence but is challenging for previously undescribed imbalances, particularly for ongoing pregnancies. Recurrent CNVs have been documented in patients with reproductive disorders; however, the application of CMA in this field is still limited. CMA enhances reproductive medicine as it facilitates better understanding of the genetic aspects of human development and reproduction and more informed patient management. Further clinical validation of CMA in the prenatal setting, creation of practice guidelines and catalogs of newly discovered submicroscopic imbalances with clinical outcomes are areas that will require attention in the future.


Effects of epidural anesthesia on incidence of venous thromboembolism following joint replacement.
Deep vein thrombosis (DVT) and pulmonary embolism are serious complications following hip and knee replacement surgery. Both surgical procedures have significantly high postoperative rates of DVT. In total hip replacement, the rate of proximal thrombi tends to be higher. However, total knee replacement produces a high rate of calf thrombi and a higher overall rate of thrombosis. In a search for innovative ways to reduce the risk of thrombosis, three types of anesthesia were examined: general anesthesia, epidural anesthesia, and hypotensive epidural anesthesia in which a controlled hypotension is provided. The hypotension commonly exists with epidural anesthesia also, unless blood pressure is maintained with an agent such as ephedrine. When blood pressure is allowed to drop in a controlled fashion, hypotensive anesthesia is effected.


Initial insights into bacterial succession during human decomposition.
Decomposition is a dynamic ecological process dependent upon many factors such as environment, climate, and bacterial, insect, and vertebrate activity in addition to intrinsic properties inherent to individual cadavers. Although largely attributed to microbial metabolism, very little is known about the bacterial basis of human decomposition. To assess the change in bacterial community structure through time, bacterial samples were collected from several sites across two cadavers placed outdoors to decompose and analyzed through 454 pyrosequencing and analysis of variable regions 3-5 of the bacterial 16S ribosomal RNA (16S rRNA) gene. Each cadaver was characterized by a change in bacterial community structure for all sites sampled as time, and decomposition, progressed. Bacteria community structure is variable at placement and before purge for all body sites. At bloat and purge and until tissues began to dehydrate or were removed, bacteria associated with flies, such as Ignatzschineria and Wohlfahrtimonas, were common. After dehydration and skeletonization, bacteria associated with soil, such as Acinetobacter, were common at most body sites sampled. However, more cadavers sampled through multiple seasons are necessary to assess major trends in bacterial succession.


An epidemiologic investigation of alcohol-dependent schizophrenics.
Most clinicians agree that alcoholism is frequent in schizophrenic patients. However, little is known about the clinical or familial characteristics of this group. We compared alcoholic schizophrenics and nonalcoholic schizophrenics with respect to sociodemographic and clinical characteristics and the rate of various psychiatric illnesses among their first-degree relatives. The only difference in the sociodemographic characteristics was a higher proportion of males among the alcoholic schizophrenics. Clinically, the alcoholic schizophrenics were more likely to report experiencing hallucinations, depressive episodes, manic episodes (females only) and multiple substance abuse (males only). In addition, the male alcoholic schizophrenics were younger at first hospitalization than the male nonalcoholic schizophrenics. This effect was reversed for females. The relatives of alcoholic schizophrenics were 2.6 times more likely to be alcoholic than the relatives of the nonalcoholic schizophrenics. The morbidity for other psychiatric disorders was similar in the two groups.


Green leaf volatiles, fire and nonanoic acid activate MAPkinases in the model grass species Lolium temulentum.
Previously it has been shown that mechanical wounding, salinity and heat activated a 46 kDa and 44 kDa mitogen-activated protein kinases (MAPKs) in forage related grasses. Forage and turf related grasses are utilized in diverse environments where they are routinely subjected to herbicides and exposed to fire and volatiles after cutting, however very little is known concerning the perception or molecular responses to these different stresses or compounds. In the model grass species Lolium temulentum (Lt), a 46 kDa mitogen-activated protein kinase (MAPK) was activated in the leaves within 5 min and a 44 kDa MAPK 15 min after exposure to green leaf volatiles released from grass clippings. When the tips of leaves of Lt plants were scorched by fire, the 46 kDa MAPK and 44 kDa MAPK were rapidly activated within 5 min and 20 min respectively in the treated leaf, and 15 min systemically in an adjacent untreated tiller after exposure to fire. Nonanoic acid (pelargonic acid), a component in herbicides used on grasses, activated a 46 kDa MAPK in the treated leaves within 5 min of exposure and 15 min in systemic tissues. At concentrations normally used in the herbicides, nonanoic acid was found to only weakly activate the 44 kDa MAPK after an hour in treated leaves, but strongly activated it in the systemic tillers 30 min after treatment. Acetic acid, HCl and NaOH also were found to activate these MAPKs in treated tillers. The rapid activation of these MAPKs to a wide range of stress stimuli, suggest that these MAPKs play a role in the perception and response to these stresses and compounds. The activation of the MAPK by green leaf volatiles indicates a role for these compounds in wound signaling in grasses.


Detection and Classification of Diabetic Macular Edema with a Desktop-Based Code-Free Machine Learning Tool.
To evaluate the effectiveness of the Lobe application, a machine learning (ML) tool that can be used on a personal computer without requiring coding expertise, in the recognition and classification of diabetic macular edema (DME) in spectral-domain optical coherence tomography (SD-OCT) scans. A total of 695 cross-sectional SD-OCT images from 336 patients with DME and 200 OCT images of 200 healthy controls were included. Images with DME were classified into three main types: diffuse retinal edema (DRE), cystoid macular edema (CME), and cystoid macular degeneration (CMD). To develop the ML model, we used the desktop-based code-free Lobe application, which includes a pre-trained ResNet-50 V2 convolutional neural network and is available free of charge. The performance of the trained model in recognizing and classifying DME was evaluated with 41 DRE, 28 CMD, 70 CME, and 40 normal SD-OCT images that were not used in the training. The developed model showed 99.28% sensitivity and 100% specificity for class-independent detection of DME. Sensitivity and specificity by labels were 87.80% and 98.57% for DRE, 96.43% and 99.29% for CME, and 95.71% and 95.41% for CMD, respectively. To our knowledge, this is the first evaluation of the effectiveness of Lobe with ophthalmological images, and the results indicate that it can be used with high efficiency in the recognition and classification of DME from SD-OCT images by ophthalmologists without coding expertise.


1-Benzoyl-3-methyl-3-pentyl-thio-urea.
Two independent mol-ecules comprise the asymmetric unit of the title compound, C(14)H(20)N(2)OS. These differ in the relative orientations of the pentyl chains [C-C-C-C torsion angles = -176.7 (3) and 176.4 (3)°]. Significant twists are evident in each mol-ecule, the dihedral angles formed between the thio-urea and amide residues being 53.47 (17) and 55.81 (17)°. In the crystal, each mol-ecule self-associates via a centrosymmetric eight-membered {⋯HNC=S}(2) synthon, and these are connected into a supra-molecular chain along [110] via C-H⋯O contacts. Disorder is noted for one of the independent mol-ecules in that two orientations (50:50) were resolved for its benzene ring.


Characterizing Single Polymeric and Protein Nanoparticles with Surface Plasmon Resonance Imaging Measurements.
Near-infrared surface plasmon resonance imaging (SPRI) microscopy is used to detect and characterize the adsorption of single polymeric and protein nanoparticles (PPNPs) onto chemically modified gold thin films in real time. The single-nanoparticle SPRI responses, Δ%RNP, from several hundred adsorbed nanoparticles are collected in a single SPRI adsorption measurement. Analysis of Δ%RNP frequency distribution histograms is used to provide information on the size, material content, and interparticle interactions of the PPNPs. Examples include the measurement of log-normal Δ%RNP distributions for mixtures of polystyrene nanoparticles, the quantitation of bioaffinity uptake into and aggregation of porous NIPAm-based (N-isopropylacrylamide) hydrogel nanoparticles specifically engineered to bind peptides and proteins, and the characterization of the negative single-nanoparticle SPRI response and log-normal Δ%RNP distributions obtained for three different types of genetically encoded gas-filled protein nanostructures derived from bacteria.


Heteroleptic dipyrrin/bipyridine complexes of ruthenium(II).
The synthesis and characterization of a series of heteroleptic dipyrrinato/2,2'-bipyridine complexes of ruthenium(II) are reported. Spectroscopic analysis, including resonance Raman, indicates that the complexes are only weakly emissive and that the dipyrrin and Ru --> bipyridine (metal-to-ligand charge transfer) chromophores are uncoupled.


Treatment of outflow tract obstruction due to benign prostatic hyperplasia with the pollen extract, cernilton. A double-blind, placebo-controlled study.
Whilst prostatectomy remains the "gold standard" for the treatment of outflow tract obstruction due to benign prostatic hyperplasia, medical treatment--if only for symptomatic relief--appears to be an attractive alternative. Most of the pharmacological agents in use block the hormonal or the sympathetic neurological pathways that influence prostate growth and function. All of these drugs are known to have side effects. Sixty patients with outflow obstruction due to benign prostatic hyperplasia (BPH) were entered into a double-blind, placebo-controlled study to evaluate the effect of a 6-month course of the pollen extract, Cernilton. There was a statistically significant subjective improvement with Cernilton (69% of the patients) compared with placebo (30%). There was a significant decrease in residual urine in the patients treated with Cernilton and in the antero-posterior (A-P) diameter of the prostate on ultrasound. However, differences in respect of flow rate and voided volume were not statistically significant. It is concluded that Cernilton has a beneficial effect in BPH and may have a place in the treatment of patients with mild or moderate symptoms of outflow obstruction.


Image-Based Food Classification and Volume Estimation for Dietary Assessment: A Review.
A daily dietary assessment method named 24-hour dietary recall has commonly been used in nutritional epidemiology studies to capture detailed information of the food eaten by the participants to help understand their dietary behaviour. However, in this self-reporting technique, the food types and the portion size reported highly depends on users' subjective judgement which may lead to a biased and inaccurate dietary analysis result. As a result, a variety of visual-based dietary assessment approaches have been proposed recently. While these methods show promises in tackling issues in nutritional epidemiology studies, several challenges and forthcoming opportunities, as detailed in this study, still exist. This study provides an overview of computing algorithms, mathematical models and methodologies used in the field of image-based dietary assessment. It also provides a comprehensive comparison of the state of the art approaches in food recognition and volume/weight estimation in terms of their processing speed, model accuracy, efficiency and constraints. It will be followed by a discussion on deep learning method and its efficacy in dietary assessment. After a comprehensive exploration, we found that integrated dietary assessment systems combining with different approaches could be the potential solution to tackling the challenges in accurate dietary intake assessment.


Cardiac lymph and contractility of the heart.
The transport mechanism of cardiac lymph with reference to the contractility of the heart was studied experimentally using a dye technique. An augmentation of cardiac contractility with or without hypoxia of the heart equally accelerated the transportation of cardiac lymph. The short staining time of the "cardiac node" correlated well to the augmented contractility of the heart and the long to the decreased contractility. Similar result was obtained in the dogs with surgical block and pacing at a rate of 100/min. This experiment suggests the importance of dynamical observation of cardiac lymph.


An examination of the moderating role of growth mindset in the relation between social stress and externalizing behaviors among adolescents.
Experiencing elevated stress increases the risk of further difficulties in mental health, including externalizing behaviors in adolescents. There is a need for understanding the factors that help adolescents mitigate social stress to prevent problematic externalizing behaviors. Growth mindset has been found to be a promising protective factor in adolescent development including mental health. This study aimed to examine the potential buffering role of growth mindset of thoughts-emotion-and-behaviors in the context of two types of social stress (i.e., family and peer) to reduce externalizing behaviors among adolescents. The sample is composed of 399 adolescents (age range 14-18 years, M = 16.22, SD = 1.21; 56.4% female, 42.3% male, 1.3% gender-nonconforming or variant) from a large, urban, diverse high school in the United States. Adolescent self-reports were administered at one time. Moderation analyses revealed that growth mindset of thoughts, emotion, and behaviors moderate the association between family stress and externalizing behaviors. The magnitude of this association weakened as the level of growth mindset increased, supporting the buffering effect of the moderator. Neither the moderation effect of growth mindset nor the main effects of peer stress and growth mindset were significant in predicting externalizing behaviors. Taken together, the findings suggested that growth mindset has protective effects in reducing externalizing behaviors when adolescents experience family stress. Future directions and implications are discussed.


[Long-term effect of acupuncture on quality of life in patients with early stage of stroke].
To study the long-term effects of acupuncture on quality of life in patients with early stage of stroke. Eighty patients were randomly assigned to the acupuncture group and the control group, 40 in each. All received conventional Western medical treatment, but for patients in the acupuncture group, acupuncture was applied additionally immediately from 48 h after patients' life signs were stabilized. Curative effects were evaluated with scores of SF-36, function independent measure (FIM) and NIHSS. Effect evaluated immediately after acupuncture operation showed that all the scores in the acupuncture group, except score of recognition function in FIM, and scores of body pain and physical function in SF-36, were improved more significantly than those in the control group (P < 0.05). The effectiveness improved continuously in the 6-month period after stroke in both groups, but more superiority was shown in the acupuncture group in aspects of emotional function, social function and mental health by SF-36 (P < 0.05). Early applying acupuncture could raise the physical, social, and psychological functions of stroke patients. Although the effects might decrease to some extent later, acupuncture still showed markedly effect on patients' psychological and social functions as well as on their life satisfaction.


Effect of rapeseed meal supplementation to gestation diet on reproductive performance, blood profiles and milk composition of sows.
This experiment evaluated the effect of dietary supplementation levels of rapeseed meal (RSM) in gestation diets on reproductive performance, blood profiles, milk composition of sows, and growth of their progeny. A total of 55 mixed-parity sows (Yorkshire×Landrace; average parity = 3.82) with an initial body weight (BW) of 193.0 kg were used in this experiment. Sows were allotted to one of 5 treatments at breeding based on BW and backfat thickness in a completely randomized design. Treatments consisted of dietary RSM supplementation levels (0%, 3%, 6%, 9%, and 12%) in gestation diets. During lactation all sows were fed a common lactation diet with no RSM supplementation. Body weight, backfat thickness, litter size, lactation feed intake, and milk composition of sows, and growth of their progeny were not different among dietary treatments. In blood profiles, a quadratic increase (Quadratic, p<0.05) in serum triiodothyronine (T3) concentration and a linear increase (Linear, p<0.01) in serum thyroxine (T4) concentration were observed at d 110 of gestation as dietary RSM supplementation levels increased. However, serum T3 and T4 concentrations in lactating sows and their piglets were not affected by RSM supplementation of gestation diets. Concentrations of serum total cholesterol and low density lipoprotein cholesterol in sows were not influenced by dietary treatments, whereas serum glucose level in sows decreased linearly at d 110 of gestation (Linear, p<0.05) by increasing dietary RSM supplementation in gestation diets. The RSM could be supplemented to gestation diets up to 12% with no detrimental effects on reproductive performance and growth of their progeny. However, increasing supplementation levels of RSM in gestation diets may increase serum T3 and T4 concentrations and decrease serum glucose concentration of sows in late gestation.


Connective tissue activation by synovial fluids and synovial-tissue extracts of rheumatoid arthritis patients.
Prompted by Castor's investigations (4) on a connective-tissue-activating peptide (CTAP) we investigated the stimulative effect of synovial fluids and synovial-tissue extracts on the synthesis of collagen by incubated embryonic-chick tendon cells. The stimulative effect was greater with synovial-tissue extracts from more severe cases than with samples from patients having milder forms of the disease. There was no correlation between the stimulation by synovial fluids and synovial-tissue extracts from the same patient. The stimulative activity was lost at dialysis. A slight stimulation in the incorporation of glucosamine was also observed. Treatment of the patients with gold, chloroquine or steroids decreased the stimulating capacity. These effects seem to depend on factors different from those described by Castor. The differerences in the stimulant activity of samples from various groups cannot be entirely due to glutamine, which is one of the limiting nutrients of the embryonic-chick tendon cells.


Solubility of Pharmaceutical Ingredients in Natural Edible Oils.
Natural edible oils (NEOs) are common excipients for lipid-based formulations. Many of them are complex mixtures comprising hundreds of different triglycerides (TGs). One major challenge in developing lipid-based formulations is the variety in NEO compositions affecting the solubility of active pharmaceutical ingredients. In this work, solubilities of indomethacin (IND), ibuprofen (IBU), and fenofibrate (FFB) in soybean oil and in coconut oil were measured via differential scanning calorimetry, high-performance liquid chromatography, and Raman spectroscopy. Furthermore, this work proposes an approach that mimics NEOs using one key TG and models the API solubilities in these NEOs based on perturbed-chain statistical associating fluid theory (PC-SAFT). Key TGs were determined using the 1,2,3-random hypothesis, and PC-SAFT parameters were estimated via a group-contribution method. Using the proposed approach, the solubility of IBU and FFB was modeled in soybean oil and coconut oil. Furthermore, the solubilities of five more APIs (IND, cinnarizine, naproxen, griseofulvin, and felodipine) were modeled in soybean oil. All modeling results were found in very good agreement with the experimental data. The influence of different NEO kinds on API solubility was examined by comparing FFB and IBU solubilities in soybean oil and refined coconut oil. PC-SAFT was thus found to allow assessing the batch-to-batch consistency of NEO batches in silico.


Urolithiasis in HIV-positive patients treated with atazanavir.
Among protease inhibitors, atazanavir has not been associated with urolithiasis in clinical studies. We describe 11 cases of atazanavir-associated urolithiasis in patients with human immunodeficiency virus (HIV) infection. Patients with low water intake, high urinary pH, and a prior history of urinary stones may have a higher risk of atazanavir-associated urine crystallization.


gp330 and RAP: the Heymann nephritis antigenic complex.
It is apparent that significant progress has been made in the characterization of gp330 in the years that have elapsed since its initial identification as the nephritogenic antigen of Heymann nephritis. However, there are still many gaps in our knowledge and we do not yet have a full picture of the molecular events leading to the formation of immune deposits in glomerular capillaries. Moreover, we still do not have direct information on the normal function(s) of gp330 and RAP and their trafficking in renal and other epithelia. The availability of the yolk sac and other cell lines that express gp330 and RAP together with the identification of the functional domains of RAP should greatly facilitate experimental studies designed to elucidate these problems. Progress will also be greatly facilitated in the future when the complete amino-acid sequence of gp330 becomes available, making possible further structural studies. It is our hope that new knowledge obtained on the molecular mechanisms of HN will provide insights into the molecular pathogenesis of human membranous nephropathy and will provide a strategy for the design of appropriate treatments to interrupt the process.


Tracking of serum lipoprotein (a) concentration and its contribution to serum cholesterol values in children from 7 to 36 months of age in the STRIP Baby Study. Special Turku Coronary Risk Factor Intervention Project for Babies.
We investigated the tracking phenomenon of serum lipoprotein (a) concentrations and assessed the impact of serum concentration of lipoprotein (a) cholesterol on total cholesterol concentrations in children from 7 to 36 months of age. Serum samples for lipoprotein (a) and cholesterol determinations at 7, 13, 24 and 36 months were prospectively obtained from 430 children. Serum lipoprotein (a) was determined using immunoradiometric assay. A strong correlation was observed between lipoprotein (a) concentrations at 7 and 36 months of age (r = 0.88, P < 0.001). Seventy-eight per cent to 86% of the children in the lowest and highest lipoprotein (a) quintiles at 13 months remained in the respective quintiles at 36 months. The average contribution of lipoprotein (a) cholesterol to total cholesterol varied from 0.5% to 3.2% (individual variation 0.13-32.39%) depending on the type of milk received and the age of the children. At 7 months the contribution was 0.44% in breast-fed and 0.93% in formula-fed infants (P < 0.0001). The tracking phenomenon of serum lipoprotein (a) concentrations is strong already in early childhood. The contribution of lipoprotein (a) cholesterol to serum total cholesterol concentration should be taken into account when the changes in serum cholesterol levels are interpreted in the first year of life.


Aldehyde Oxidase Mediated Metabolism in Drug-like Molecules: A Combined Computational and Experimental Study.
Aldehyde oxidase (AOX) is an important drug-metabolizing enzyme. However, the current in vitro models for evaluating AOX metabolism are sometimes misleading, and preclinical animal models generally fail to predict human AOX-mediated metabolism. In this study, we report a combined computational and experimental investigation of drug-like molecules that are potential aldehyde oxidase substrates, of which multiple sites of metabolism (SOMs) mediated by AOX and their preferences for the reaction can be unambiguously identified. In addition, the proposed strategy was used to evaluate the metabolism of newly designed c-Met inhibitors, and a success switch-off of AOX metabolism was observed. Overall, this study provide useful information to guide lead optimization and drug discovery based on AOX-mediated metabolism.


[Robotic cardiac surgery].
Cardiac surgery has evolved towards less traumatic procedures. Minimal invasive cardiac surgery is a well-established technic offering patients the same level of excellence accomplished with open surgery. Technological evolution has lead to robotic assistance for minimal invasive cardiac surgery, helping surgeons to perform even better in cardiac repairs or reconstructions.


Pyrethroid resistance associated with M918 L mutation and detoxifying metabolism in Aphis gossypii from Bt cotton growing regions of China.
The cotton aphid, Aphis gossypii (Glover), is a destructive pest that affects numerous crops throughout the world. Pyrethroid resistance has become endemic in A. gossypii populations in the cotton growing areas of China due to wide- spread application of insecticides. To assess the extent of pyrethroid resistance, bioassays were conducted on field populations collected from several cotton cultivation areas from 2010 to 2015. The frequency of a known resistance-associated sodium channel mutation (M918 L) in A. gossypii was evaluated and the bioassay of bifenthrin with or without the synergist was performed to illuminate the mechanisms underlying resistance to pyrethroids. The field populations exhibited very high levels of resistance to both beta-cypermethrin and deltamethrin. Pretreatment with synergists, DEF and PBO, significantly increased the toxicity of bifenthrin to cotton aphid populations collected from Bt cotton fields in China. Further, 96.8-100% of individuals with the M918 L mutation (including both RR and RS individuals) were observed in various populations, and only 2.8-3.2% of individuals with wild-type homozygotes (SS) were detected. The mutation M918 L in the voltage-gated sodium channel along with detoxifying metabolism was contributed to the pyrethroid resistance in the field populations of Aphis gossypii from cotton growing regions of China. And insecticides with different modes of action should be recommended for the control of A. gossypii in the future. © 2017 Society of Chemical Industry.


Interaction between chlorhexidine digluconate and EDTA.
The combination of chlorhexidine and EDTA produces a white precipitate. The aim of this study was to determine if the precipitate involves the chemical degradation of chlorhexidine. The precipitate was produced and redissolved in a known amount of dilute trifluoroacetic acid. The amount of chlorhexidine and EDTA present in the dissolved precipitate was determined by reverse-phase high performance liquid chromatography (HPLC) with ultraviolet detection at 288 nm. More than 90% of the precipitate's mass was found to be EDTA or chlorhexidine. The remainder is suspected to be water, gluconate, and sodium. Parachloroaniline, a potentially carcinogenic decomposition product of chlorhexidine, was not detected in the precipitate (the limit of detection was 1%). The molar ratio of chlorhexidine to EDTA in the precipitate was about 1.6 to 1. Based on the results, chlorhexidine forms a salt with EDTA rather than undergoing a chemical reaction.


Core-Shell-Structured Electrorheological Fluid with a Polarizability-Tunable Nanocarbon Shell for Enhanced Stimuli-Responsive Activity.
The incorporation of nanocarbon-based materials into electrorheological fluids has been shown to be an effective means of improving the electrorheological (ER) response. However, the mechanism of the sp2/sp3-hybridized carbon structure and high ER response is still under investigation. Herein, barium titanate@nanocarbon shell (BTO@NCs) composites are proposed and prepared by introducing carbonized polydopamine (C-PDA) into a shell. When the polymerization time of dopamine is tuned, the shell thickness, surface polar functional groups, and sp2/sp3-hybridized carbon can be effectively controlled. The maximum yield stress of the BTO@NCs-24 h ER fluid reaches 2.5 kPa under an electric field of 4 kV mm-1, which is attributed to the increased content of sp3 C-OH and oxygenous functional groups within the shell, resulting in a rapidly achievable polarization. Furthermore, the SiO2@NCs and TiO2@NCs ER fluids are also prepared with enhanced ER behavior in these phenomena, confirming an approach to high-performance ER fluids based on nanocarbon composites.


Thermal safety considerations for implantable micro-coil design.
Micro magnetic stimulation of the brain via implantable micro-coils is a promising novel technology for neuromodulation. Careful consideration of the thermodynamic profile of such devices is necessary for effective and safe designs.Objective.We seek to quantify the thermal profile of bent wire micro-coils in order to understand and mitigate thermal impacts of micro-coil stimulation.Approach. In this study, we use fine wire thermocouples and COMSOL finite element modeling to examine the profile of the thermal gradients generated near bent wire micro-coils submerged in a water bath during stimulation. We tested a range of stimulation parameters previously reported in the literature such as voltage amplitude, stimulus frequency, stimulus repetition rate and coil wire materials.Main results. We found temperature increases ranging from <1 °C to 8.4 °C depending upon the stimulation parameters tested and coil wire materials used. Numerical modeling of the thermodynamics identified hot spots of the highest temperatures along the micro-coil contributing to the thermal gradients and demonstrated that these thermal gradients can be mitigated by the choice of wire conductor material and construction geometry.Significance. ISO standard 14708-1 designates a thermal safety limit of 2 °C temperature increase for active implantable medical devices. By switching the coil wire material from platinum/iridium to gold, our study achieved a 5-6-fold decrease in the thermal impact of coil stimulation. The thermal gradients generated from the gold wire coil were measured below the 2 °C safety limit for all stimulation parameters tested.


Nucleotide sequence of the leukotoxin genes of Pasteurella haemolytica A1.
A 4.4-kilobase-pair DNA fragment coding for the leukotoxin of Pasteurella haemolytica A1 has been isolated, and its nucleotide sequence has been determined. Two open reading frames, designated lktC and lktA, coding for proteins of 19.8 and 101.9 kilodaltons, respectively, were identified. Expression of the two genes in minicell-labeling experiments resulted in the production of the predicted proteins LKTC and LKTA. By using an antiserum against the soluble antigens of P. haemolytica A1 in Western blot (immunoblot) analysis of total cellular proteins from the Escherichia coli clones, LKTA was identified as an additional antigenic protein. Results from subcloning of the DNA fragment suggested that expression from both lktC and lktA is required for leukotoxin activity, indicating that the leukotoxin of P. haemolytica A1 is encoded by two genes. A comparison of the organization and the DNA sequence of the leukotoxin genes with those of the E. coli alpha-hemolysin genes showed a significant degree of homology between the two loci. This analysis suggested that the leukotoxin genes of P. haemolytica A1 and the E. coli alpha-hemolysin genes may have evolved from a common ancestor and that the two toxins may share similar activities or functional domains or both.


PROGNOSTIC FACTORS IN PATIENTS WITH PRESSURE SORES IN A UNIVERSITY HOSPITAL IN SOUTHERN BRAZIL.
Despite advances in medical care, patients who are hospitalized or have spinal cord injuries often develop pressure sores. The objective of this study was to describe the epidemiological characteristics of pressure sores and evaluate factors associated with recurrence and cure. In this historical cohort study, clinical and laboratory data were collected from medical records between 1997 and 2016. Sixty individuals with pressure ulcers were included; mean patient age was 38.1±16.5 (37.0) years, 83.3% were men, and 86.8% identified as white. Most patients (85.1%) had paraplegia, amputation, or trauma of the lower limbs with motor sequelae; the remainder (14.9%) were quadriplegic. Most (78.3%) underwent surgery, and the mean follow-up time was 1.8±2.5 years. The lesions were cured in 25 patients; they recurred in 25% of the patients, and recurrence was seen to be associated with the location of the lesions. Patients with recurrent lesions had more medical consultations and a longer treatment time. Individuals whose ulcers had healed had fewer lesions, higher body mass index (BMI), and a higher proportion of these patients underwent surgery. BMI and location and number of lesions are prognostic factors. Level of Evidence IV, Case Series.


Replication, recombination and packaging of amplicon DNA in cells infected with the herpes simplex virus type 1 alkaline nuclease null mutant ambUL12.
The alkaline nuclease (AN) encoded by gene UL12 of herpes simplex virus type 1 (HSV-1) is essential for efficient virus replication but its role during the lytic cycle remains incompletely understood. Inactivation of the UL12 gene results in reductions in viral DNA synthesis, DNA packaging, egress of DNA-containing capsids from the nucleus and ability of progeny virions to initiate new cycles of infection. Mechanistically, AN has been implicated in resolving branched structures in HSV-1 replicative intermediates prior to encapsidation, and promoting DNA strand-exchange. In this study, amplicons (bacterial plasmids containing functional copies of a virus replication origin and packaging signal) were used to analyse further the defects of the UL12 null mutant ambUL12. When ambUL12 was used as a helper virus both replication and packaging of the transfected amplicon were reduced in comparison with cells infected with wild-type (wt) HSV-1, and to extents similar to those previously observed for genomic ambUL12 DNA. By using amplicons differing at a specific restriction endonuclease site it was demonstrated that replicating molecules exhibit high frequency intermolecular recombination in both wt- and mutant-infected cells. Surprisingly, in the absence of the UL12 product, amplicons lacking a functional encapsidation signal were packaged. Moreover, these packaged molecules could be serially propagated indicating that they had been incorporated into functional virions. This difference in packaging specificity between wt HSV-1 and ambUL12 might indicate that replicative intermediates accumulating in the absence of AN contain an increased incidence of structures that can serve for the initiation of DNA packaging.


Unspecific binding capacity of androgen receptors in the rat uterus.
Rat uterine cytosol receptor homogenate binds estradiol and testosterone alike. Saturation and displacement studies were done with these two hormones. Testosterone displaced only testosterone while estradiol displaced estradiol as well as testosterone. The experiments call attention to the unspecific binding capacity of rat uterine androgen receptors.


Design of Cationic Lipids with Acetal Linkers: Conformational Preferences, Hydrolytic Stability, and High Drug-Loading Abilities.
Lipids are key constituents of numerous biomedical drug delivery technologies. Here, we present the design, synthesis and biophysical characterizations of a library of cationic lipids containing an acetal residue in their linker region. These cationic acetal lipids (CALs) were conveniently prepared through a trans-acetalization protocol from commercially available precursors. NMR studies highlighted the conformational rigidity at the acetal residue and the high hydrolytic stability of these CALs. Fluorescence anisotropy studies revealed that the CAL with a pyridinium headgroup (CAL1) formed highly cohesive vesicular aggregates in water. These structural and self-assembly features of the CAL1 allowed up to 196 % w/w loading of curcumin (Cur) as a representative hydrophobic drug. A reconstitutable formulation of Cur was obtained as a result, which could deliver the drug inside mammalian cells with very high efficiency. The hemocompatibility and cytocompatibility of CAL1 was significantly enhanced by creating a coating of polydopamine (PDA) onto its vesicular assemblies to produce hybrid lipid-polymer nanocapsules. This work demonstrates rapid access to the useful synthetic lipid formulations with high potential in drug and gene delivery applications.


Subcellular distribution of cyclic adenosine 3',5'-monophosphate-binding protein and estrogen receptors in control pituitaries and estrogen-induced pituitary tumors.
Binding of cyclic adenosine 3',5'-monophosphate (cAMP) and steroid receptors was studied in cytoplasmic and nuclear fractions of pituitaries from castrated rats, in rats subjected to acute (60 min) or short-term (4 days) estradiol (E2) treatment, and in diethylstilbestrol-induced pituitary tumors (DES-T). E2 receptors were primarily in nuclear extracts in all animals that were given estrogens, whereas cytosolic receptors were low to absent. Contrarily, castrated rats showed high quantities of cytoplasmic receptor but little in nuclear sites. The progestin receptor was induced only in 4-day E2-treated rats and in DES-T. cAMP binding was stimulated in cytosol from 4-day E2-treated rats and in DES-T, but a significant reduction in binding was also noted in nuclear extracts from DES-T. Scatchard analysis for the cytosolic cAMP-binding activity demonstrated a two-component system, and the increased cAMP binding obtained in DES-T seemed to be caused by an increase in the low-affinity, high-capacity binder [regulatory type II (RII) subunit of protein kinase]. Suggestion of the preferential estrogenic induction of RII was also obtained by DEAE-cellulose chromatography, which provided separation of RI and RII subunits. The results suggest that sustained estrogenization leads to induction of cytosolic cAMP-binding protein and increased levels of nuclear E2 receptor. In DES-T, this effect resulted in an inverse subcellular distribution of both binding proteins, which may be related to abnormal growth of the pituitary, as has been postulated for hormone-dependent mammary tumors.


Relapse of Wegener's granulomatosis in the first trimester of pregnancy: a case report.
The association of Wegener's granulomatosis and pregnancy is rare and poses unique therapeutic challenges, particularly when active disease presents in early pregnancy. We describe a 22-yr-old woman who recovered successfully from her initial episode of Wegener's granulomatosis with a standard course of treatment with prednisolone and cyclophosphamide. Two and a half years later, she presented with relapse during the first trimester of pregnancy (primigravida). Since the clinical features suggested mild disease, she was started on prednisolone at a dose of 1 mg/kg/day, to which she seemed to respond very well for 3 months. Unfortunately, she had a spontaneous abortion at 5 months of gestation while on 25 mg/day of prednisolone. At this time, her disease flared further, with clinically manifest lung disease which was not part of her initial presentation. She was treated with another course of oral cyclophosphamide and prednisolone, and a remission was achieved in 4 months. There are no agreed guidelines on the treatment of Wegener's granulomatosis during pregnancy. In this report, the therapeutic issues are discussed against the background of the available literature.


The effect of physiotherapy on standing balance in chronic anterior cruciate ligament insufficiency.
Body sway movements in the frontal plane in a single-limbed stance test were used to assess postural control in 26 patients with chronic anterior cruciate ligament insufficiency. The injured and the noninjured legs were tested before the patients were committed to physiotherapy for 3 to 6 months. Follow-up tests were made after 3, 12, and 36 months. Significant disturbance of the balance of both legs was found before training, compared with a reference group of normal subjects. Values of the noninjured leg were normalized after 3 months of training, but the injured leg still showed an increased body sway. Normal balance parameters on the injured side were found at examination after 12 months. Follow-up examination after 36 months proved persistent normalization of the single-limbed balance on both sides.


Effectiveness of a Skin Care Program With a Cream Containing Ceramide C and a Personalized Training for Secondary Prevention of Hand Contact Dermatitis.
Background/Objectives: The aim of our study was to investigate the effectiveness of personalized training on skin protection associated with the regular use of ceramide-containing cream (CC) versus other creams (OC) for improving hand contact dermatitis. Methods: We performed a double-center randomized trial that enrolled workers with hand dermatitis. All workers received personalized training. The intervention was 3 times per day application of the study emollient. The control arm used an emollient of choice without ceramide, as needed. The primary outcome was improvement in hand dermatitis at 1 and 3 months of follow-up. Results: In total, 102 patients with hand dermatitis were enrolled in this study. Improvement in dermatitis was found in 40%, 52.5%, 50%, and 63% of OC and CC, at the first and second follow-ups, respectively. The use of CC was significantly associated with an improvement in dermatitis (odds ratios 2.6; 95% confidence intervals 1.30-5.2), analyzed using generalized equation estimation during the follow-up. Conclusion: Our study demonstrated that an educational personalized intervention could improve the signs and symptoms in patients with hand dermatitis, and the use of a CC resulted in a significantly better outcome during the 3 months of follow-up.


Management of sudden sensorineural hearing loss - a national survey.
The management of Sudden Sensorineural Hearing loss is a controversial issue and there is no universally agreed protocol in its management The aim of this study was to ascertain the referral trends, treatment strategies, and outcome of treatment adopted by the Consultant Otorhinolarygologists in this country. A postal questionnaire was sent to all 32 Consultants in the Republic of Ireland, which achieved 21(65.62%) responses. Patients were referred to 13(61.9%) respondents within two weeks of the onset of symptoms and 3(14.28%) Consultants were referred patients within twenty four hours of the onset of symptoms. All specialists routinely performed full blood count and erythrocyte sedimentation rate, 15(71.42%) routinely assess fasting blood sugar and 5(23.80%) investigate fasting lipids and perform a syphilis screen. MR imaging to outrule vestibular schwanoma is performed by 11(52.38%) respondents. When treated within two weeks of the symptoms 6(28.57%) of the respondents were optimistic about recovery of hearing upon treatment whereas 2(9.52%) considered recovery of useful hearing less likely if treated more than two weeks after onset The triple therapy with oral vasodilators, oral steroids, and Carbogen inhalation was favoured by 9(42.85%). Oral steroid was the sole therapeutic modality by 4(19.04%). The opinion of Consultants was that the referral trend from the primary care physician is appropriate and 20(95.23%). Consultants considered an optimistic outcome if treated within two weeks. The investigations and management protocol for sudden sensorineural hearing loss is not uniform which is consistent with the treatment strategies world wide.


Radiation therapy dose and androgen deprivation therapy in localized prostate cancer: a meta-regression of 5-year outcomes in phase III randomized controlled trials.
While multiple randomized trials have evaluated the benefit of radiation therapy (RT) dose escalation and the use and prolongation of androgen deprivation therapy (ADT) in the treatment of prostate cancer, few studies have evaluated the relative benefit of either form of treatment intensification with each other. Many trials have included treatment strategies that incorporate either high or low dose RT, or short-term or long-term ADT (STADT or LTADT), in one or more trial arms. We sought to compare different forms of treatment intensification of RT in the context of localized prostate cancer. Using preferred reporting items for systemic reviews and meta-analyses (PRISMA) guidelines, we collected over 40 phases III clinical trials comparing different forms of RT for localized prostate cancer. We performed a meta-regression of 40 individual trials with 21,429 total patients to allow a comparison of the rates and cumulative proportions of 5-year overall survival (OS), prostate cancer-specific mortality (PCSM), and distant metastasis (DM) for each treatment arm of every trial. Dose-escalation either in the absence or presence of STADT failed to significantly improve any 5-year outcome. In contrast, adding LTADT to low dose RT significantly improved 5-year PCSM (Odds ratio [OR] 0.34, 95% confidence interval [CI] 0.22-0.54, p < 0.001) and DM (OR 0.35, 95% CI 0.20-0.63. p < 0.001) over low dose RT alone. Adding STADT also significantly improved 5-year PCSM over low dose RT alone (OR 0.55, 95% CI 0.41-0.75, p < 0.001). While limited by between-study heterogeneity and a lack of individual patient data, this meta-analysis suggests that adding ADT, versus increasing RT dose alone, offers a more consistent improvement in clinical endpoints.


The role of calcium in the initiation of fast axonal transport.
Incubation of neuronal cell bodies in a calcium-free medium depresses the amount, but not the rate, of fast axonal transport of [3H]protein. Under these conditions, which do not affect protein synthesis or general energy metabolism, less protein appears to be loaded onto the transport system. Depression of transport also is seen when cell bodies are exposed to medium containing Co2+; selective exposure of axons to this medium has no effect on transport. These findings have led to the concept of an initiation phase of fast axonal transport that comprises the events by which selected proteins are transferred from their polysomal sites of synthesis to the transport system. The divalent cation specificity of the Ca2+ requirement, and its occurrence subsequent to Golgi apparatus-associated glycosylation, suggest that proteins destined for fast axonal transport are routed through the soma in a manner similar to that of secretory proteins and integral membrane proteins in nonneural cells. This analogy is pursued to consider a scheme whereby Golgi-derived vesicles deliver fast-transported proteins to the axonal smooth endoplasmic reticulum. Possible roles of Ca2+ in the formation and exocytotic fusion of such vesicles are considered.


Proteomic profiling uncovered the cytosolic superoxide dismutase BsSOD1 associated with plant defence in the herbal orchid Bletilla striata.
The herbal orchid Bletilla striata (Thunb.) Rchb.f. has a long cultivation history and has been widely used in medicines and cosmetics. The fungal infection leaf blight (LB) seriously threatens B. striata cultivation. Here, we systemically collected wild B. striata accessions and isolated the accessions with strong resistance against LB. We carried out proteomic profiling analysis of LB-resistant and LB-susceptible accessions, and identified a large number of differentially expressed proteins with significant gene ontology enrichment for 'oxidoreductase activity.' Of the proteins identified in the reactive oxygen species signalling pathway, the protein abundance of the Cu-Zn superoxide dismutase BsSOD1 and its gene expression level were higher in LB-resistant accessions than in LB-susceptible lines. Transient expression of the dismutase fused with yellow fluorescent protein determined that its subcellular localisation is in the cytoplasm. Our study provides new insights into the molecular markers associated with fungal infection in B. striata.


Gut Microbial Diversity Assessment of Indian Type-2-Diabetics Reveals Alterations in Eubacteria, Archaea, and Eukaryotes.
Diabetes in India has distinct genetic, nutritional, developmental and socio-economic aspects; owing to the fact that changes in gut microbiota are associated with diabetes, we employed semiconductor-based sequencing to characterize gut microbiota of diabetic subjects from this region. We suggest consolidated dysbiosis of eubacterial, archaeal and eukaryotic components in the gut microbiota of newly diagnosed (New-DMs) and long-standing diabetic subjects (Known-DMs) compared to healthy subjects (NGTs). Increased abundance of phylum Firmicutes (p = 0.010) and Operational Taxonomic Units (OTUs) of Lactobacillus (p < 0.01) were observed in Known-DMs subjects along with the concomitant graded decrease in butyrate-producing bacterial families like Ruminococcaceae and Lachnospiraceae. Eukaryotes and fungi were the least affected components in these subjects but archaea, except Methanobrevibacter were significantly decreased in them. The two dominant archaea viz. Methanobrevibacater and Methanosphaera followed opposite trends in abundance from NGTs to Known-DMs subjects. There was a substantial reduction in eubacteria, with a noticeable decrease in Bacteroidetes phylum (p = 0.098) and an increased abundance of fungi in New-DMs subjects. Likewise, opportunistic fungal pathogens such as Aspergillus, Candida were found to be enriched in New-DMs subjects. Analysis of eubacterial interaction network revealed disease-state specific patterns of ecological interactions, suggesting the distinct behavior of individual components of eubacteria in response to the disease. PERMANOVA test indicated that the eubacterial component was associated with diabetes-related risk factors like high triglyceride (p = 0.05), low HDL (p = 0.03), and waist-to-hip ratio (p = 0.02). Metagenomic imputation of eubacteria depict deficiencies of various essential functions such as carbohydrate metabolism, amino acid metabolism etc. in New-DMs subjects. Results presented here shows that in diabetes, microbial dysbiosis may not be just limited to eubacteria. Due to the inter-linked metabolic interactions among the eubacteria, archaea and eukarya in the gut, it may extend into other two domains leading to trans-domain dysbiosis in microbiota. Our results thus contribute to and expand the identification of biomarkers in diabetes.


Gene expression profiling of circulating CD133+ cells of hepatocellular carcinoma patients associated with HCV infection.
Identifying the genetic expression profile of CD133+ cells from HCC patients compared to CD133+ cells from healthy volunteers that may contribute in hepatocarcinogenesis process. Circulating CD133+ cells were sorted from the peripheral blood of HCC patients as well as from healthy volunteers using magnetic activated cell sorting. The differential expression profile of stem cell related genes was performed using the Stem Cell PCR profiling assay. Data analysis of stem cells related genes in CD133+ cells of the HCC group compared to the control group showed that; CCND2, COL1A1, CTNNA1, DLL3, JAG1, KRT15, MYC, NOTCH2, T and TERT were up-regulated (fold change=80, 68.6, 6.67, 7.22, 3.8, 15.2, 14.5, 105.6, 26.6 and 99 respectively while only CD3D was down-regulated (fold change=0.055) in HCC patients. However, after application of Beferroni correction to adjust P-value; KRT15 was the only gene that was significantly over expressed in CD133+ cells of HCC compared to control group (P-value=0.012). KRT15 can be used to differentiate between circulating CD133+ cells from HCC group and control group. However, further study may be needed to confirm on the protein level.


Physicochemical characterization of Remsima.
Remsima (infliximab) was recently approved as the world's first biosimilar monoclonal antibody (mAb) in both the European Union and Korea. To achieve this, extensive physicochemical characterization of Remsima in relation to Remicade was conducted in order to demonstrate the highly similar properties between the two molecules. A multitude of state-of-the-art analyses revealed that Remsima has identical primary as well as indistinguishable higher order structures compared with the original product. Monomer and aggregate contents of Remsima were also found to be comparable with those of Remicade. In terms of charge isoforms, although Remsima was observed to contain slightly less basic variants than the original antibody, the difference was shown to be largely due to the presence of C-terminal lysine. On the other hand, this lysine was found to be rapidly clipped inside serum in vitro and in vivo, suggesting it has no effect on the biological potency or safety of the drug. Analysis of the glycan contents of the antibodies showed comparable glycan types and distributions. Recent results of clinical studies have further confirmed that the two antibody products are highly similar to each other. Based on this research as well as previous clinical and non-clinical comparability studies, Remsima can be considered as a highly similar molecule to Remicade in terms of physicochemical properties, efficacy, and safety for its final approval as a biosimilar product to Remicade.


Optimizing Mask Ventilation: Literature Review and Development of a Conceptual Framework.
Mask ventilation is lifesaving, especially in cases of difficult intubation. Many publications have offered distinct techniques for optimizing mask ventilation. This article reviews currently available difficult mask ventilation literature and theory. We divide difficult mask ventilation into 3 broad categories based on etiology: inadequate mask seal, increased airway resistance, and decreased respiratory compliance. Published strategies for overcoming difficulty are presented and organized by etiology.


Embouchure Dysfunction in Air Force Band Brass Musicians.
Occupational injuries and medical problems in musicians are well described, but relatively less attention has been paid to orofacial and embouchure-related problems in professional brass players. This study addressed embouchure-related problems in Air Force Band members, a population of musicians with an intense practice and performance schedule. A survey was developed and distributed via the Air Force Survey Office to 599 active-duty Air Force Band members and 201 Air National Guard members. The survey assessed practice patterns, practice and performance venues, and presence of symptoms suggesting embouchure dysfunction. Responses were obtained from 167 Air Force Band brass players. Of the 157 responding to the question about embouchure dysfunction, 42% reported having experienced an embouchure problem at some point in the past, and 53% of those respondents reported that they were currently experiencing an embouchure problem. Forty-one percent of those with embouchure problems cited practice venues that were not conducive to effective and efficient practice at the time their embouchure problems began, and 48% of those with embouchure problems reported having to overblow in rehearsal at the time their problems began. Embouchure disorders were reported in a large proportion of Air Force Band brass survey respondents, and specific concerns related to practice venues and the need to overblow in practice settings suggest factors suitable to remediation and preventive strategies.