Ozone containment through selective mitigation measures on precursors of volatile organic compounds.
Abatement of volatile organic compounds (VOCs) ozone reduction is usually carried out by reducing the total amount of VOCs without considering reactivity between different species. This study incorporates the concept of maximum incremental reactivity (MIR) and speciation profiles into the industrial emission inventory of Taiwan to target organic species from industrial sources with the greatest ozone formation potentials (OFPs). These high OFP sources/species are then mitigated to assess the O3 reduction amount (ΔO3) with Community Multiscale Air Quality (CMAQ) modeling under VOC-limited conditions. The objective is to minimize the number of target sources/species and their tonnage while achieving maximum O3 reduction. This approach is referred to as the Selective Precursor Mitigation (SPM). A case study of a high ozone episode (September 4-10, 2020) was chosen for illustration, during which a relatively stagnant atmospheric condition with minimal transboundary ozone occurred. A series of scenarios to target the highest OFP chemicals/industries for mitigation are compared for the achievable max. ΔO3, areas affected (area coverage), and reduction efficiency. For instance, by reducing the ten leading industry classes with the island's highest OFPs (OFPind), up to 19 % of max. 1-h ΔO3 can be expected. If, however, the same tonnage of VOCs as that of OFPind is distributed to all industries without considering the reactivity, called the overall mitigation (OM), comparable results to those of OFPind were found, but the number of sources needed to be managed with OM would increase by nearly three times (29,662 for OM vs. 11,981 for OFPind). Further reducing the management scale by only zooming in the ten highest OFP chemicals within the ten leading OFP industries (OFPsp) would result in relatively limited area coverage. Still, major ozone hot spots could be alleviated. Although the domain is set on the island of Taiwan, the SPM approach is universally applicable to other regions worldwide to gain the maximum ozone reduction effect at a minimized societal cost.


Identification and architecture of a putative secretion tube across mycobacterial outer envelope.
Tuberculosis-causing mycobacteria have thick cell-wall and capsule layers that are formed from complex structures. Protein secretion across these barriers depends on a specialized protein secretion system, but none has been reported. We show that Mycobacterium tuberculosis Rv3705c and its homologous MSMEG_6251 in Mycobacterium smegmatis are tube-forming proteins in the mycobacterial envelope (TiME). Crystallographic and cryo-EM structures of these two proteins show that both proteins form rotationally symmetric rings. Two layers of TiME rings pack together in a tail-to-tail manner into a ring-shaped complex, which, in turn, stacks together to form tubes. M. smegmatis TiME was detected mainly in the cell wall and capsule. Knocking out the TiME gene markedly decreased the amount of secreted protein in the M. smegmatis culture medium, and expression of this gene in knocked-out strain partially restored the level of secreted protein. Our structure and functional data thus suggest that TiME forms a protein transport tube across the mycobacterial outer envelope.


Ascorbic acid inhibition of cytochrome P450-catalyzed uroporphyrin accumulation.
Previous studies on the mechanism of the uroporphyria caused by polyhalogenated aromatic hydrocarbons have indicated a key role of cytochrome P450 of the 1A subfamily in catalyzing uroporphyrinogen (UROgen) oxidation. Here we report that ascorbic acid (ASC) inhibits UROgen oxidation in primary cultures of chick embryo hepatocytes and hepatic microsomes from chickens and mice. In hepatocyte cultures, 0.15 mM ASC totally prevented the accumulation of uroporphyrin (URO) induced by treatment of cells with the combination of 3,4,3',4'-tetrachlorobiphenyl (TCB) and 2-propyl-2-isopropylacetamide (PIA), but had no effect on the induction of protoporphyrin accumulation by PIA and desferrioxamine. However, addition of 5-aminolevulinic acid (ALA) to cultures treated with PIA plus TCB decreased the ability of ASC to prevent URO accumulation, suggesting that the effectiveness of ASC was dependent on the intracellular concentration of ALA or its metabolites. Similarly, when chick hepatocyte cultures were treated with TCB plus exogenous ALA to produce URO accumulation, the effectiveness of ASC was also less than when ALA was produced endogenously. Under this condition, addition of piperonyl butoxide, a P450 inhibitor, increased ASC inhibition of URO accumulation. ASC competitively inhibited the oxidation of UROgen by hepatic microsomes from chicks or mice treated with 3-methylcholanthrene (MC) with Ki for ASC being about 0.1 mM. ASC prevented formation of a 500-nm absorbing compound, probably tetrahydrouroporphyrin, the first intermediate in UROgen oxidation. These results are consistent with ASC preventing URO accumulation in hepatocytes by competitive inhibition of the first step of UROgen oxidation and suggest a new physiological role of ASC, that of maintaining UROgen in the reduced state.


Chronic Wounds: Innovations in Diagnostics and Therapeutics.
A major global health issue is the existence of chronic wounds. Appropriate diagnosis and treatment is essential to promote wound healing and prevent further complications. Traditional methods for treatment and diagnosis of chronic wounds have shown to be of limited effectiveness. Therefore, there is a need for the development of diagnostic and therapeutic innovations in chronic wound care. This mini-review aims to provide insight in the current knowledge of the wound healing process and the deficiencies encountered in chronic wounds, which provides a basis for the development of innovations in chronic wound care. Furthermore, promising diagnostic and therapeutic innovations will be highlighted. Literature was searched for recent articles (=<10 years) describing the current knowledge about the wound healing process and chronic wounds. The most promising diagnostic and therapeutic innovations were gathered from articles published in the past 5 years. Wound healing is a well-organized process consisting of four phases: coagulation, inflammation, proliferation and wound remodelling. Chronic wounds often stagnate in the inflammatory phase and/or experience an impaired proliferative phase. This mini-review has demonstrated that increased knowledge about the processes involved in wound healing has paved the way for the development of new diagnostic tools and treatments for chronic wounds. Increased knowledge about bacterial invasion and infection in has encouraged researchers to develop diagnostic tools to help clinicians detect these phenomena appropriately and in time. Other researchers have shown that they are able to design/extract biochemical compounds that intervene in the disrupted healing processes in chronic wounds.


Fluorescent staining of nuclear envelope coated with heparin.
Naturally and artificially devitalized cells of various origin can be loaded with heparin and subsequently stained with DAPI, a newer fluorochrome (4',6-diamidino-2-phenylindole), leading to a bright yellow fluorescence of the nuclear envelope. Depending on the cell type it is also possible to coat the outer cell membrane with heparin or other sulphated mucopolysaccharides and to achieve similar staining phenomena with DAPI.


Obese mouse fat cell-derived extracellular vesicles transport miR-99a-5p to mitigate the proliferation and migration of non-small cell lung cancer cells.
Fat cells-derived extracellular vesicles (FC-EVs) play a role in regulating the tumor microenvironment in cancers by transporting RNAs. MicroRNAs (miRNAs) are vital regulators of cancer development. This study was conducted to explore the role of FC-EVs in the proliferation and migration of non-small cell lung cancer (NSCLC) cells, providing targets for NSCLC treatment. The obese mouse model was established via high-fat diet (HFD), followed by separation and characterization of FC-EVs (HFD-EVs). The levels of miR-99a-5p, precursor-miR-99a-5p, and heparan sulfate-glucosamine 3-sulfotransferase 3B1 (HS3ST3B1) were measured by RT-qPCR or Western blot assay. Cell proliferation and migration were evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide and wound healing assays. The expression of Cy3-labeled miR-99a-5p in A549 cells (one NSCLC cell line) was observed via confocal microscopy. The binding of miR-99a-5p to HS3ST3B1 was analyzed by the dual luciferase assay. Rescue experiments were performed to confirm the role of HS3ST3B1 in NSCLC cells. miR-99a-5p was upregulated in adipose tissues, FCs, and HFD-EVs. HFD-EVs mitigated the proliferation and migration of NSCLC cells. HFD-EVs transported miR-99a-5p into A549 cells, which upregulated miR-99a-5p expression and inhibited HS3ST3B1 expression in A549 cells. HS3ST3B1 overexpression reversed the inhibition of HFD-EVs on the proliferation and migration of NSCLC cells. HFD-EVs transported miR-99a-5p into NSCLC cells and inhibited HS3ST3B1, thereby inhibiting proliferation and migration of NSCLC cells.


Interaction study between levofloxacin and omeprazole using urinary pharmacokinetic data.
The objective of this study was to observe the drug interaction between levofloxacin and omeprazole using urinary excretion data. Levofloxacin tablet and omeprazole capsule were administered separately as well as in combination in fasting condition with a wash out period of two weeks after each administration. Urine was collected at different time intervals of 0, 0-2, 2-4, 4-8, 8-12, 12-24, 24-36 and 36-48 hr post-dose and analyzed using a validated HPLC with UV detection. Different pharmacokinetic parameters for both drugs were determined using non-compartmental method. The maximum rate of excretion (R(max)) of levofloxacin was not decreased significantly when co-administered with omeprazole (p<0.05). Similarly no significant difference (p = 0.350) was observed for Rmax of omeprazole when co-administered with levofloxacin. Again the fraction of levofloxacin excreted (f(e)/f) was not changed significantly (p = 0.953) due to the co-administration of omeprazole. Similarly fraction of omeprazole excreted (f(e)/f) also remained unaffected (p = 0.672) when co-administered with levofloxacin. No significant change was observed for the area under the rate of excretion versus midpoint of time interval curve from zero to 48 hours (AURC(0-48)) for levofloxacin and omeprazole (p = 0.816 and 0.792 respectively) when administered separately and co-administered with each other. The study clearly revealed that levofloxacin and omeprazole do not undergo any kind of interactions when administered together. So it can be concluded that these two drugs can be prescribed together to achieve optimum therapeutic activity.


[Allergic granulomatosis and angiitis associated with intestinal perforation and eye movement disorder].
A perforation in the small intestine and one and a half syndrome were observed in 45-year-old male with allergic granulomatosis and angiitis (AGA). The patient was first admitted to our hospital in 1988 with complaints of bronchial asthma and an abnormal density on his chest X ray. The diagnosis of PIE syndrome was made based on marked peripheral blood eosinophilia. He was successfully treated with prednisolone and had been well on the maintenance therapy of small doses of it. In May 1990, peripheral neuropathy occurred in his bilateral lower legs and it was followed by nausea and vomiting. The laboratory findings on admission showed leukocytosis (19,300/mm3) with 43% eosinophilia and elevated blood level of IgE (3000 IU/ml). On the tenth day of admission, ileus symptom and subsequently a perforation of the small intestine were observed. The mesenterial specimens obtained during the emergency operation showed the findings compatible with AGA. Remission of AGA was induced by the treatment with 60 mg prednisolone and 100 mg cyclophosphamide per day following methylprednisolone pulse therapy. He was well on maintenance prednisolone until June 1991, when he suddenly developed one and a half syndrome. Although cranial CT scan and MRI showed no abnormal findings, cyclophosphamide therapy was resumed and prednisolone was increased because microangiopathy due to AGA was thought to be the cause of this central nervous system symptom. The one and a half syndrome improved with this therapy. This case suggests that the long term treatment with prednisolone and cyclophosphamide would be indicated in the severe AGA with close monitoring of the symptoms and peripheral blood eosinophilia.


High cervical spinal cord stimulation for unstable angina pectoris.
Twelve patients with grade IV unstable angina were treated with high cervical spinal cord stimulation (SCS). All patients had been previously treated with maximal tolerated doses of beta-blockers and calcium antagonists and had no possibility of revascularization surgery or intraluminal angioplasty. Multipolar electrodes were implanted using a percutaneous technique, and the parameters of stimulation were pulse width 0.1 ms, frequency 120 Hz, intensity that caused no unpleasant paresthesias, with two periods of 4 h 'on' daily. After a mean follow-up of 9.8 +/- 8.2 months the clinical results show a significant reduction in the number of angina attacks (p less than 0.01) and in their grade (p less than 0.001). In the treadmill ergometric test, a significant increase in the time until angina and double product (p less than 0.01) was observed. We conclude that the SCS is a very low-risk technique that significantly increases the quality of life of patients with unstable angina.


Determination of fexofenadine in tablets by capillary electrophoresis in free solution and in solution with cyclodextrins as analyte carriers.
Capillary electrophoresis (CE) methods for the determination of fexofenadine (FEX) in commercial pharmaceuticals were developed. It was demonstrated that FEX could be effectively analyzed in free solution cationic CE at low pH. Another analytical approach studied was based on cyclodextrin (CD) modified CE where highly charged CD derivatives served as analyte carriers. In this way, the separation range was spread to physiological pH region and a CE analysis of FEX, present actually in its zwitterionic form, could be accomplished. Several parameters affecting the separations were studied, including the type and concentration of carrier ion, counterion, analyte carrier, and pH of the buffer. The methods based on the free solution CE and CD-modified CE were compared each other, validated, and applied for the determination of FEX in tablets.


Coronal/Hemicoronal Approach - A Gateway to Craniomaxillofacial Region.
The coronal incision with its various modifications provides the most versatile approach to various areas in the craniomaxillofacial region coupled with excellent exposure. The aesthetic advantage of a hidden scar in the hairline, accounts for its continued popularity. The aim of this study was to review the surgical anatomy, technique and problems of post-operative morbidity pertinent to coronal approach in various clinical situations such as craniofacial trauma, tumour resections and reconstructive craniofacial procedures. In this study, ten patients who presented to Oral and maxillofacial surgery department with various craniofacial problems requiring the use of coronal and hemicoronal approach for treatment were evaluated over a period of two years. Five patients needed coronal approach and another five underwent the surgical procedures through hemicoronal approach. This was an observational study. It was observed that a well-planned and carefully designed coronal/hemicoronal incision with strict adherence to surgical principles posed minimal complication during surgery as well as post-operatively. None of the patients developed infection or heamatoma in the postoperative period. Sensory nerve deficits along the distribution of supraorbital nerve was observed in four patients of bicoronal approach and three patients of hemicoronal approach which completely resolved at the end of six months. Motor nerve weakness was observed in four patients in immediate postoperative period which gradually improved. But it persisted in one patient even after six months who had pathology of temporo-orbital region. All the patients had transient alopecia along the line of incision which improved at the end of six months. No other significant disadvantages or complications were noted. This approach offers widest accessibility and visibility to the entire upper and middle one third of the face in less than twenty minutes as observed in our study. The postoperative complications are minimal, minor and outweigh the advantages for surgical treatment in any given clinical situation as observed in this study. This proves the brilliance of coronal approach in solving an array of surgical problems pertinent to craniomaxillofacial region with superior aesthetic outcomes. ZMC-Zygomatico maxillary complex, NOE-Nasoethmoidal complex, LF-Lefort, ORIF-open reduction internal fixation.


Pulmonary complications of feeding tubes: a new technique of insertion and monitoring malposition.
To confirm the value of a new technique that will ensure safe introduction of feeding tubes. Two case reports; an anatomical and physiologic description of deglutition; and a case study. A level 2 regional referral centre. Thirteen anesthetized adult patients and 7 awake subjects, comprising patients scheduled to undergo elective surgery, medical staff and health care volunteers. Airway sampling for carbon dioxide with capnography in 13 anesthetized adults with the tip of the feeding tube in the pharynx, in the esophagus and in the trachea, and airway sampling for carbon dioxide from the pharynx and esophagus in 7 awake subjects during introduction of the feeding tube. Fluoroscopic monitoring of the position of the tip of the feeding tube during introduction in two patients and two volunteers. Carbon dioxide levels at the tip of the feeding tube during introduction. In all patients, with the tube either in the trachea or pharynx, a normal capnogram was displayed. When the tube was introduced into the esophagus no capnogram curve was seen, indicating the absence of carbon dioxide. With the subject lying down during introduction, the weighted tube followed the posterior pharyngeal wall to the upper esophageal sphincter. Positioning of the patient lying down with the head flexed and capnographic measurement of carbon dioxide levels from the tip of the feeding tube during insertion is a safe, accurate and cost-effective method for the introduction of feeding tubes.


Intraspinal epidermoid cyst.
An intraspinal epidermoid cyst occurring in the lumbo-sacral region of a 38-year old man with congenital spina bifida occulta is described. Relevant literature on the subject is reviewed.


Roles of Ileal ASBT and OSTα-OSTβ in Regulating Bile Acid Signaling.
In addition to their classical role as detergents, bile acids function as signaling molecules to regulate gastrointestinal physiology, carbohydrate and lipid metabolism, and energy expenditure. The pharmacodynamic potential of bile acids is dependent in part on the tight pharmacokinetic control of their concentration and metabolism, properties governed by their hepatic synthesis, enterohepatic cycling, and biotransformation via host and gut microbiota-catalyzed pathways. Key Messages: By altering the normal cycling and compartmentalization of bile acids, changes in hepatobiliary or intestinal transport can affect signaling and lead to the retention of cytotoxic hydrophobic bile acids and cell injury. This review discusses advances in our understanding of the intestinal transporters that maintain the enterohepatic cycling of bile acids, signaling via bile acid-activated nuclear and G protein receptors, and mechanisms of bile acid-induced cell injury. Dysregulated expression of the Asbt and Ostα-Ostβ alters bile acid signaling via the gut-liver farnesoid X receptor-fibroblast growth factor 15/19 axis and may contribute to other bile acid-regulated metabolic and cell injury pathways.


Coalescent methods are robust to the simultaneous effects of long branches and incomplete lineage sorting.
It is well known that species with elevated substitution rates can give rise to disproportionately long branches in the species tree. This combination of long and short branches can contribute to long-branch artifacts (LBA). Despite efforts to remedy LBA via increased taxon sampling and methodological improvements in gene tree estimation, it remains unclear how long and short branches affect species tree estimation in the presence of incomplete lineage sorting (ILS). Here, we examine the combined influence of long external and short internal branches on concatenation and coalescent methods using both simulated and empirical data. Our results demonstrate that the presence of long and short branches alone does not obviously confound the consistency of concatenation and coalescent methods. However, when long external and short internal branches occur simultaneously with high ILS, concatenation methods can be misled, especially when two of these long branches are sister lineages. In contrast, coalescent methods are more robust under these circumstances. This is particularly relevant because this topological pattern also characterizes numerous ancient rapid radiations across the tree of life. Because short internal branches can increase the potential for ILS and gene tree discordance, our results collectively suggest that coalescent methods are more likely to infer the correct species tree in cases of ancient rapid radiations where long external and short internal branches are in close phylogenetic proximity.


Comparison of assessment and management of suicidal risk for acute psychiatric assessment between two state sponsored hospitals in England and Italy.
The risk of suicide is one of the most important risk factors looked into for acute psychiatric assessments that influences the management plan. The prevalence of suicide is on a rise across European countries; as a consequence, the different countries have created specific guidelines and policies in order to prevent suicides in the acute settings. These guidelines are based on both different cultural aspects as well as the different organization of the mental health system in the different countries. This paper wants to present the comparison between the guidelines of two European countries, England and Italy, in order to evaluate the systems, understand differences and common contact points. The different European countries could learn one from the other and a European shared point of view may be a way forward to create better understanding and preventing the risk of suicide across the population.


Structural determination of Rickettsia lipid A without chemical extraction confirms shorter acyl chains in later-evolving Spotted Fever Group pathogens.
Rickettsiae are Gram-negative obligate intracellular parasites of numerous eukaryotes. Human pathogens of the Transitional Group (TRG), Typhus Group (TG), and Spotted Fever Group (SFG) rickettsiae infect blood-feeding arthropods, have dissimilar clinical manifestations, and possess unique genomic and morphological attributes. Lacking glycolysis, rickettsiae pilfer numerous metabolites from host cytosol to synthesize peptidoglycan and lipopolysaccharide (LPS). For LPS, O-antigen immunogenicity varies between SFG and TG pathogens; however, lipid A proinflammatory potential is unknown. We previously demonstrated that R. akari (TRG), R. typhi (TG), and R. montanensis (SFG) produce lipid A with long 2' secondary acyl chains (C16 or C18) compared to short 2' secondary acyl chains (C12) in R. rickettsii (SFG) lipid A. To further probe this structural heterogeneity and estimate a time point when shorter 2' secondary acyl chains originated, we generated lipid A structures for two additional SFG rickettsiae ( R. rhipicephali and R. parkeri ) utilizing Fast Lipid Analysis Technique adopted for use with tandem mass spectrometry (FLAT n ). FLAT n allowed analysis of lipid A structure directly from host cell-purified bacteria, providing substantial improvement over lipid A chemical extraction. FLAT n -derived structures indicate SFG rickettsiae diverging after R. rhipicephali evolved shorter 2' secondary acyl chains. Bioinformatics analysis of Rickettsia LpxL late acyltransferases revealed discrete active sites and hydrocarbon rulers for long versus short 2' secondary acyl chain addition. While the significance of different lipid A structures for diverse Rickettsia pathogens is unknown, our success using FLAT n will facilitate determining how structural heterogeneity impacts interactions with host lipid A receptors and overall inflammatory potential. Deforestation, urbanization, and homelessness lead to spikes in Rickettsioses. Vector-borne human pathogens of Transitional Group (TRG), Typhus Group (TG), and Spotted Fever Group (SFG) rickettsiae differ by clinical manifestations, immunopathology, genome composition, and morphology. We previously showed that lipid A (or endotoxin), the membrane anchor of Gram-negative bacterial lipopolysaccharide (LPS), structurally differs in R. rickettsii (later-evolving SFG) relative to R. montanensis (basal SFG), R. typhi (TG), and R. akari (TRG). As lipid A structure influences recognition potential in vertebrate LPS sensors, further assessment of Rickettsia lipid A structural heterogeneity is needed. Here, we sidestepped the difficulty of ex vivo lipid A chemical extraction by utilizing FLAT n , a new procedure for generating lipid A structures directly from host cell-purified bacteria. These data confirm later-evolving SFG pathogens synthesize structurally distinct lipid A. Our findings impact interpreting immune responses to different Rickettsia pathogens and utilizing lipid A adjuvant or anti-inflammatory properties in vaccinology.


A fully integrated standalone portable cavity ringdown breath acetone analyzer.
Breath analysis is a promising new technique for nonintrusive disease diagnosis and metabolic status monitoring. One challenging issue in using a breath biomarker for potential particular disease screening is to find a quantitative relationship between the concentration of the breath biomarker and clinical diagnostic parameters of the specific disease. In order to address this issue, we need a new instrument that is capable of conducting real-time, online breath analysis with high data throughput, so that a large scale of clinical test (more subjects) can be achieved in a short period of time. In this work, we report a fully integrated, standalone, portable analyzer based on the cavity ringdown spectroscopy technique for near-real time, online breath acetone measurements. The performance of the portable analyzer in measurements of breath acetone was interrogated and validated by using the certificated gas chromatography-mass spectrometry. The results show that this new analyzer is useful for reliable online (online introduction of a breath sample without pre-treatment) breath acetone analysis with high sensitivity (57 ppb) and high data throughput (one data per second). Subsequently, the validated breath analyzer was employed for acetone measurements in 119 human subjects under various situations. The instrument design, packaging, specifications, and future improvements were also described. From an optical ringdown cavity operated by the lab-set electronics reported previously to this fully integrated standalone new instrument, we have enabled a new scientific tool suited for large scales of breath acetone analysis and created an instrument platform that can even be adopted for study of other breath biomarkers by using different lasers and ringdown mirrors covering corresponding spectral fingerprints.


Transcriptional and epigenetic regulation of follicular T-helper cells and their role in autoimmunity.
T-follicular helper (Tfh) cells are a specialized subset of T cells that provide help to B cells and promote the formation of germinal centers (GCs). Tfh cells transmit important signals to B cells that drive class switch recombination, somatic hyper-mutation, the generation of high-affinity antibodies, immunological memory and their differentiation into plasma cells or memory B cells in the GCs. Tfh-cell differentiation is regulated by the coordinated functions of distinct cytokines, including interleukin (IL)-6, IL-21, IL-12, IL-23, IL-2, IL-7 and transforming growth factor-β (TGF-β), as well as transcription factors, including B-cell lymphoma 6 protein (Bcl-6), Signal transducers and activators of transcription (STAT)1, STAT3, STAT4, B-cell activating transcription factor (Batf), interferon regulatory factor 4 (IRF4), v-maf avian musculoaponeurotic fibrosarcoma oncogene homolog (C-Maf), T-cell-specific transcription factor 1 (TCF-1) and Achaete-scute homolog 2 (Acl2), which have been shown to form a complex transcriptional network. In addition, increasing evidence indicates that epigenetic regulations, such as DNA methylation, histone modifications and non-coding RNA regulations, also coordinately control the differentiation and function of Tfh cells along with transcription factors. Furthermore, abnormalities in the regulation of Tfh cells have been associated with several autoimmune diseases, such as systemic lupus erythematosus (SLE), multiple sclerosis (MS) and rheumatoid arthritis (RA). Herein, this review aims to summarize the coordinate regulation and interaction of transcription factors, cytokines and epigenetic modifications that control Tfh-cell differentiation as well as the mechanism of dysregulation of Tfh cells in pathogenesis of autoimmune diseases, which highlight potential therapeutic targets.


Pancreatic α Cell-Derived Glucagon-Related Peptides Are Required for β Cell Adaptation and Glucose Homeostasis.
Pancreatic α cells may process proglucagon not only to glucagon but also to glucagon-like peptide-1 (GLP-1). However, the biological relevance of paracrine GLP-1 for β cell function remains unclear. We studied effects of locally derived insulin secretagogues on β cell function and glucose homeostasis using mice with α cell ablation and with α cell-specific GLP-1 deficiency. Normally, intestinal GLP-1 compensates for the lack of α cell-derived GLP-1. However, upon aging and metabolic stress, glucose tolerance is impaired. This was partly rescued with the DPP-4 inhibitor sitagliptin, but not with glucagon administration. In isolated islets from these mice, glucose-stimulated insulin secretion was heavily impaired and exogenous GLP-1 or glucagon rescued insulin secretion. These data highlight the importance of α cell-derived GLP-1 for glucose homeostasis during metabolic stress and may impact on the clinical use of systemic GLP-1 agonists versus stabilizing local α cell-derived GLP-1 by DPP-4 inhibitors in type 2 diabetes.


[Remodeling of the left ventricle following myocardial infarction: physiopathology and prevention].
The degree of left ventricular dilatation occurring after myocardial infarction is a major prognostic factor. Left ventricular remodeling is characterized by early expansion of the infarcted region and predominantly excentric hypertrophy of the noninfarcted myocardium, resulting in left ventricular cavity dilatation. Remodeling is most pronounced in large anterior infarctions. Recent evidence indicates that remodeling is amenable to treatment. Several randomized studies have demonstrated that long-term administration of converting enzyme inhibitors not only reduces left ventricular enlargement but also lowers morbidity and mortality after myocardial infarction.


Intravesical injection of botulinum toxin type a may be an effective treatment option for autonomic dysreflexia in patients with high-level spinal cord injury.
To evaluate the efficacy of intravesical injection of botulinum toxin type A (BTX-A) for neurogenic detrusor overactivity (DO) in reducing the frequency and severity of autonomic dysreflexia (AD). A cross-sectional nonrandomized trial with before (baseline) and after (follow-up) assessments. A single spinal cord injury (SCI) rehabilitation center in China. Twenty-five patients with SCI at or above T6 and a history of AD who underwent urodynamic studies (UDS) before and 3 months after BTX-A injection. Received bladder injection treatment wtih 200 U BTX-A. The maximum detrusor pressure(Pdetmax) and voume at first DO(VFIDC), baseline and overall maximum systolic blood pressure (SBP) during UDS, and scores of Incontinence Specific Quality of Life Instrument (IQoL) were recorded before and 3 months after the injection. The change in SBP (ΔSBP) from baseline to maximum SBP during UDS was calculated to assess the severity. The frequency of AD was recorded using ambulatory blood pressure monitoring during a 24 h period before and 3 months after the injection. BTX-A injection decreased the Pdetmax and increased the VFIDC and mean urine volume per catheterization increased. The maximum SBP and the ΔSBP during UDS decreased significantly decreased after the injection (151.44 ± 13.92 vs 133.32 ± 9.20 mmHg and 49.44 ± 12.81 vs 33.08 ± 9.11 mmHg respectively, P < 0.05). The frequency of bladder-related ADs (i.e. performed a clean intermittent catheterization or leakage) during a 24-h period significantly decreased from 11.04 ± 1.81-7.88 ± 2.15 (P < 0.001). BTX-A decreases the severity of SBP increase and the number of AD episodes 3 months after intravesical injection.


Regulation of hepatic cytochrome P-450 during infectious disease.
During episodes of infectious disease the mixed function oxidase system is depressed and the capacity of the liver to metabolize drugs can be compromised in both animals and humans. The depression that occurs during viral infections is mediated via the production of interferon. This action of interferon requires the synthesis of an intermediate protein(s) yet to be identified. Using an oligonucleotide probe for a unique sequence in cytochrome P-450LA omega we have now shown that the mRNA for this isozyme is depressed following the administration of interferon inducers. The magnitude in the loss of mRNA corresponds to the magnitude of the loss in the levels of this isozyme. This depression is observed within 6 h of interferon exposure. It is concluded that the decrease in drug metabolism during viral infections is caused by an interferon-mediated loss in mRNA and subsequent cytochrome P-450 synthesis in the liver.


Energy transfer among dyes on particulate solids.
Absorption and fluorescence properties of methylene blue (MB), a well-known singlet molecular oxygen photosensitizer, and its mixtures with pheophorbide-a (Pheo) sorbed on microgranular cellulose are studied, with emphasis on radiative and nonradiative energy transfer from Pheo to MB. Although pure MB builds up dimeric species on cellulose even at 2 x 10(-8) mol g(-1), addition of 2.05 x 10(-7) mol g(-1) Pheo largely inhibits aggregation up to nearly 10(-6) mol g(-1) MB. At the same time, the absorption spectrum of monomeric MB in the presence of Pheo differs from the spectrum in pure cellulose. Both effects reveal a strong influence of Pheo on the medium properties. A model relying entirely on experimental data is developed, through which energy transfer efficiencies can be calculated for thin and thick layers of dye-loaded cellulose. At the largest concentration of MB assuring no dye aggregation, nonradiative energy transfer efficiencies reach a maximum value of nearly 40%. This value is quite high, taking into account the low fluorescence quantum yield of Pheo, Phi = 0.21, and results from the existence of high local concentrations of the acceptor within the supporting material. These results show that large energy transfer rates can exist in a system devoid of any special molecular organization.


[CONSTRUCTION OF TISSUE ENGINEERED COMPOSITE WITH THERMOSENSITIVE COLLAGEN HYDROGEL IN DYNAMIC CULTURE SYSTEM].
To explore the morphological and functional features of tissue engineered composite constructed with bone mesenchymal stem cells (BMSCs) as seeding cells, thermosensitive collagen hydrogel (TCH) and poly-L-lactic acid (PLLA) as the extracellular matrix (ECM) scaffolds in the dynamic culture system. BMSCs were separated from long bones of Fischer 344 rat, and cultured; and BMSCs at the 3rd generation were seeded on the ECM scaffold constructed with braided PLLA fiber and TCH. The BMSCs-ECM scaffold composite was cultured in the dynamic culture system which was designed by using an oscillating device at a frequency of 0.5 Hz and at swing angle of 70 degrees (experimental group), and in the static culture system (control group) for 7 days. The general observation and scanning electron microscopy (SEM) observation were performed; total DNA content was.measured at 0, 1, 3, and 7 days. PLLA was surrounded by collagen to form translucent gelatiniform in 2 groups; and compact membrane developed on the surface of PLLA. SEM observation showed that BMSCs had high viability and were fusiform in shape with microvilli on the surface of cells, and arranged in line; collagen and cells filled in the pores of PLLA fiber in the experimental group. The cells displayed a flat shape on the surface; there were less cells filling in the pores of PLLA fiber in the control group. At 1, 3, and 7 days, total DNA content in the experimental group was significantly higher than that in control group (P < 0.05). The total DNA content were increased gradually with time in 2 groups, showing significant difference between at 0 day and at 7 days (P < 0.05). The ECM constructed with TCH and PLLA has good biocompatibility. The dynamic cultivation system can promote the cell proliferation, distribution, and alignment on the surface of the composite, so it can be used for tissue engineered composite in vitro.


Ethological studies in the ant Tetramorium caespitum Mayr. II. Interspecific relationships.
The structure of relationships between the neighboring species of ants is characterized by great variability. What happens when particular individuals meet - depends upon considerable differences between individual behaviors of ants and a number of variable factors determining the actual emotional state of the workers. From these factors, it is the general structure of forces between the nests that changes slowest, and may be responsible for reaching temporary stabilization of the interspecific relationships. Invariable hierarchical structure between the species occurs very rarely: when there is an enormous supremacy of one of the species.


Overexpression of pulmonary extracellular superoxide dismutase attenuates endotoxin-induced acute lung injury.
Superoxide is produced by activated neutrophils during the inflammatory response to stimuli such as endotoxin, can directly or indirectly injure host cells, and has been implicated in the pathogenesis of acute lung injury (ALI)/acute respiratory distress syndrome (ARDS). We wished to determine the potential for pulmonary overexpression of the extracellular isoform of superoxide dismutase (EC-SOD) to reduce the severity of endotoxin-induced lung injury. Animals were randomly allocated to undergo intratracheal instillation of (1) surfactant alone (vehicle); (2) adeno-associated virus (AAV) vectors containing a null transgene (AAV-null); and (3) adeno-associated virus vectors containing the EC-SOD transgene (AAV-EC-SOD) and endotoxin was subsequently administered intratracheally. Two additional groups were randomized to receive (1) vehicle or (2) AAV-EC-SOD, and to undergo sham (vehicle) injury. The severity of the lung injury was assessed in all animals 24 h later. Endotoxin produced a severe lung injury compared to sham injury. The AAV vector encoding EC-SOD increased lung EC-SOD concentrations, and enhanced the antioxidant capacity of the lung. EC-SOD overexpression decreased the severity of endotoxin-induced ALI, reducing the decrement in systemic oxygenation and lung compliance, decreasing lung permeability and decreasing histologic injury. EC-SOD attenuated pulmonary inflammation, decreased bronchoalveolar lavage neutrophil counts, and reduced interleukin-6 and CINC-1 concentrations. The AAV vector itself did not contribute to inflammation or to lung injury. Pulmonary overexpression of EC-SOD protects the lung against endotoxin-induced ALI.


Severe epistaxis after nasogastric tube insertion requiring arterial embolisation.
A 53-year-old dialysis patient was admitted with symptoms of a respiratory tract infection, abdominal pain and vomiting. She aspirated and required intubation. A nasogastric tube was placed with slight difficulty and the patient developed severe epistaxis. The bleeding could not be controlled with mechanical pressure and nasal packing. Angiography revealed extravasation from a pseudoaneurysm arising from the inferior pharyngeal branch of the ascending pharyngeal trunk. The vessel was successfully embolised with cessation of bleeding. We emphasise that even a seemingly easy procedure like insertion of a nasogastric tube, can lead to a life-threatening complication.


Effects of Lipid Lowering Drugs on Arterial Stiffness: One More Way to Reduce Cardiovascular Risk?
Arterial stiffness (AS) is considered an independent predictor of cardiovascular disease (CVD) events. Among lipid lowering drugs, statins have a beneficial effect on AS, independent of their hypolipidaemic effect. Based on 3 meta-analyses and other studies, this effect is compound- and doserelated. Potent statins at high doses are more effective than less powerful statins. Ezetimibe (± statin) also seems to decrease AS in patients with dyslipidaemia. Fibrates have no effect on AS. Proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors have data that beneficially affect all AS risk factors, suggesting a beneficial effect on artery compliance. However, there is no direct measurement of their effect on AS indices. In patients with dyslipidaemia, prescribing high dose statins (± ezetimibe) will not only decrease low-density lipoprotein cholesterol levels but also improve AS (in addition to other effects). This effect on AS may contribute to the observed reduction in vascular events.


Integrating the life course into MCH service delivery: from theory to practice.
To describe the efforts of a community-based maternal and child health coalition to integrate the life course into its planning and programs, as well as implementation challenges and results of these activities. Jacksonville-Duval County has historically had infant mortality rates that are significantly higher than state and national rates, particularly among its African American population. In an effort to address this disparity, the Northeast Florida Healthy Start Coalition embraced the life course approach as a model. This model was adopted as a framework for (1) community needs assessment and planning; (2) delivery of direct services, including case management, education and support in the Magnolia Project, its federal Healthy Start program; (3) development of community collaborations, education and awareness; and, (4) advocacy and grass roots leadership development. Implementation experience as well as challenges in transforming traditional approaches to delivering maternal and child health services are described. Operationalizing the life course approach required the Coalition to think differently about risks, levels of intervention and the way services are organized and delivered. The organization set the stage by using the life course as a framework for its required local planning and needs assessments. Based on these assessments, the content of case management and other key services provided by our federal Healthy Start program was modified to address not only health behaviors but also underlying social determinants and community factors. Individual interventions were augmented with group activities to build interdependence among participants, increasing social capital. More meaningful inter-agency collaboration that moved beyond the usual referral relationships were developed to better address participants' needs. And finally, strategies to cultivate participant advocacy and community leadership skills, were implemented to promote social change at the neighborhood-level. Transforming traditional approaches to delivering maternal and child health services and sustaining change is a long and laborious process. The Coalition has taken the first steps; but its efforts are far from complete. Based on the agency's initial implementation experience, three areas presented particular challenges: staff, resources and evaluation. The life course is an important addition to the MCH toolbox. Community-based MCH programs should assess how a life course approach can be incorporated into existing programs to broaden their focus, and, potentially, their impact on health disparities and birth outcomes. Some areas to consider include planning and needs assessment, direct service delivery, inter-agency collaboration, and community leadership development. Continued disparities for people of color, despite medical advances, demand new interventions that purposefully address social inequities and promote advocacy among groups that bear a disproportionate burden of infant mortality. Successful transformation of current approaches requires investment in staff training to garner buy-in, flexible resources and the development of new metrics to measure the impact of the life course approach on individual and programmatic outcomes.


Indirect Popliteal Artery Transections in Revision Total Knee Arthroplasty A Case Report.
Popliteal artery injury in total knee arthroplasty (TKA) is rare but a devastating complication with consequence from transient bleeding to death. We report a case of 83-yearold woman suffering from an unusual early postoperative indirect popliteal artery transection in revision TKA for an infected knee. This injury was triggered by surgical correction of stiffened and distorted soft tissue, which resulted in excessive tensioning and eventual indirect transection of the artery. To avoid this devastating complication, early elimination of infection concomitant with perioperative maintenance of the normal anatomy of the knee is important.


Faecal incontinence after anal dilatation.
We have studied 10 patients with faecal incontinence occurring after anal dilatation. Abnormalities were found in the external and sphincter in clinical and manometric tests in five patients and in single fibre EMG tests in four patients. Internal anal sphincter function was impaired in eight patients. These findings have implications for prevention of faecal incontinence after this procedure.


Functional relevance of radiographic spinal progression in axial spondyloarthritis: results from the GErman SPondyloarthritis Inception Cohort.
The aim of the study was to investigate the functional relevance of the development of structural damage in the spine in patients with early axial spondyloarthritis (axSpA). Altogether, 210 patients with early axSpA (symptom duration ⩽10 years) who completed a 2-year clinical and radiographic follow-up in the GErman SPondyloarthritis Inception Cohort were included. An association between structural damage in the spine [modified Stoke AS Spine Score (mSASSS)] and functional status (the BASFI) or spinal mobility (the BASMI) was assessed in a longitudinal linear mixed model analysis; both unstandardized (β) and standardized (βstand) regression coefficients were calculated. There was an association between mSASSS and BASFI: β = 0.05 (95% CI: 0.03, 0.08) and βstand = 0.20 (95% CI: 0.11, 0.59) adjusted for disease activity parameters (the BASDAI and CRP), the presence of definite radiographic sacroiliitis and sex. An association between mSASSS and BASMI was stronger: β = 0.08 (95% CI: 0.05, 0.11) and βstand = 0.41 (95% CI: 0.25, 0.57) adjusted for the same parameters. These data indicate that, over time, an increase of 20 or 12 mSASSS points would be responsible for an increase of one BASFI or one BASMI point, respectively. Disease activity (BASDAI) also showed a significant association with BASFI [β = 0.79 (95% CI: 0.71, 0.86) and βstand = 0.71 (95% CI: 0.63, 0.77)] and BASMI [β = 0.22 (95% CI: 0.15, 0.30) and βstand = 0.28 (95% CI: 0.18, 0.37)]. Structural damage in the spine and disease activity are both determinants of the functional status and spinal mobility in early axSpA.


Histomorphological Study on Number of Acini of the Prostate Gland of Bangladeshi Cadaver.
The prostate is made up of 30 to 50 compound tubuloalveolar glands that are embedded in a framework of fibromuscular tissue and arranged in three concentric groups (mucosal, submucosal and main prostatic gland). The prostate consists of peripheral, central and transitional zone accounting for approximately 70%, 20% and 5% of the glandular substances. Benign prostatic hyperplasia affects the transitional zone and carcinoma of the prostate affects the peripheral zone. The glandular tissue consists of numerous acini with frequent internal papillae. Follicular epithelium is variable but predominantly columnar and either single-layered or pseudostratified. This cross sectional descriptive study was conducted in Department of Anatomy, Mymensingh Medical College, Mymensingh, Bangladesh to find out the difference in number of acini of the prostate gland of Bangladeshi people in relation to age. The present study was performed on 67 postmortem human prostate gland collected from the morgue in the Department of Forensic Medicine, Mymensingh Medical College by non random purposive sampling technique. The specimens were collected from Bangladeshi cadaver of age ranging from 10 to 80 years. All the specimens were grouped into three categories: Group A (up to 18 years), Group B (19 to 45 years) and Group C (above 45 years) according to age. Dissection was performed following standard autopsy techniques. In the present study, total 60 slides were made for histological study from both central and peripheral zone of the prostate which were examined under low power objectives. The number of the acini of the prostate gland were counted and recorded. The mean number of the acini of the prostate gland was 16.45, 43.54 and 42.45 in Group A, B and C respectively in central zone and 30.08, 51.35 and 44.16 in Group A, B and C respectively in peripheral zone of the prostate. Variance analysis shows that mean differences were highly significant between Group A & B and Group C & A and non significant between Group B & C in central zone of the prostate. Mean differences were non significant among all three groups in peripheral zone of the prostate. The number of the acini of prostate gland was increased up to certain age limit but decreased in extreme age. In statistical analysis, differences between age groups were analyzed by using students unpaired 't' test. The present study will help to increase the information pool on the number of the acini of the prostate gland of Bangladeshi people at different age group.


Impact of Alcohol on Glycemic Control and Insulin Action.
Alcohol has profound effects on tissue and whole-body fuel metabolism which contribute to the increased morbidity and mortality in individuals with alcohol use disorder. This review focuses on the glucose metabolic effects of alcohol, primarily in the muscle, liver and adipose tissue, under basal postabsorptive conditions and in response to insulin stimulation. While there is a relatively extensive literature in this area, results are often discordant and extrapolating between models and tissues is fraught with uncertainty. Comparisons between data generated in experimental cell and animals systems will be contrasted with that obtained from human subjects as often times results differ. Further, the nutritional status is also an important component of the sometimes divergent findings pertaining to the effects of alcohol on the regulation of insulin and glucose metabolism. This work is relevant as the contribution of alcohol intake to the development or exacerbation of type 2 diabetes remains ill-defined and a multi-systems approach is likely needed as both alcohol and diabetes affect multiple targets within the body.


The natural history of humeral intracondylar fissure: an observational study of 30 dogs.
To determine the risk of condylar fracture, or of needing to have a transcondylar screw placed, and to identify risk factors in a cohort of dogs with humeral intracondylar fissure (also known as incomplete ossification of the humeral condyle) that was initially managed non-surgically. A retrospective owner survey of dogs diagnosed with humeral intracondylar fissure as an incidental finding and managed non-surgically with a minimum of two years follow-up. Body weight, age, estimated fissure size, gender and contralateral fracture at the time of diagnosis were evaluated as potential risk factors for the development of a humeral condylar fracture or for having a transcondylar screw placed. Data were available for 30 dogs (34 elbows). Six humeral condyles with a mean fissure size of 50% fractured at a mean of 14 months after diagnosis. A transcondylar screw was placed across two humeral condyles with fissure sizes of 60 and 100% at 11 and 17 months. No risk factors were identified for fracture/screw placement. For those cases that did not fracture or have a screw placed mean fissure size was 52% and mean follow-up time was 56 months (range 29 to 79 months). Eighteen percent of cases progressed to fracture and 24% in total required surgery. This information allows clinicians and owners to make an informed decision regarding surgery when faced with a dog with humeral intracondylar fissure identified as an incidental finding.


Neuro-Immunity Controls Obesity-Induced Pain.
The prevalence of obesity skyrocketed over the past decades to become a significant public health problem. Obesity is recognized as a low-grade inflammatory disease and is linked with several comorbidities such as diabetes, circulatory disease, common neurodegenerative diseases, as well as chronic pain. Adipocytes are a major neuroendocrine organ that continually, and systemically, releases pro-inflammatory factors. While the exact mechanisms driving obesity-induced pain remain poorly defined, nociceptor hypersensitivity may result from the systemic state of inflammation characteristic of obesity as well as weight surplus-induced mechanical stress. Obesity and pain also share various genetic mutations, lifestyle risk factors, and metabolic pathways. For instance, fat pads are often found hyper-innervated and rich in immune cell types of multiple origins. These immunocytes release cytokines, amplifying nociceptor function, which, in turn, via locally released neuropeptides, sustain immunocytes' function. Here, we posit that along with mechanical stress stemming from extra weight, the local neuro-immune interplay occurring within the fat pads maintains the state of chronic low-grade inflammation and heightens sensory hypersensitivity. Overall, stopping such harmful neuro-immune crosstalk may constitute a novel pathway to prevent obesity-associated comorbidities, including neuronal hypersensitivity.


Urinary balantidiasis: diagnosis at a glance by urine sediment examination.
A 56-year-old Caucasian man with non-Hodgkin's lymphoma, who had previously been treated with prolonged intensive chemotherapy, was hospitalized for an acute and reversible kidney injury of multifactorial origin. The urinary sediment examination, performed daily, demonstrated the presence of renal tubular cells and renal tubular cell casts. Surprisingly, it also showed the presence of trophozoites of the protozoan Balantidium coli, which were identified on the basis of its characteristic morphology and rapid movements across the slide, and transient leukocyturia. The patient was asymptomatic, his medical history was negative for gastrointestinal disease, and no Balantidium coli was found in the feces. In spite of this, due to the previous chemotherapy, the patient was treated with oral metrodinazole. Only one other case with Balantidium coli in the urine sediment has been described so far and this paper stresses the importance of the examination of the urinary sediment.


Aerosol-based efficient delivery of azithromycin to alveolar macrophages for treatment of respiratory infections.
The efficacy of aerosol-based delivery of azithromycin (AZM) for the treatment of respiratory infections caused by pathogenic microorganisms infected in alveolar macrophages (AMs) was evaluated by comparison with oral administration. The aerosol formulation of AZM (0.2 mg/kg) was administered to rat lungs using a Liquid MicroSprayer(®). The oral formulation of AZM (50 mg/kg) was used for comparison. Time-courses of concentrations of AZM in AMs following administration were obtained, and then the therapeutic availability (TA) was calculated. In addition, the area under the concentrations of AZM in AMs - time curve/minimum inhibitory concentration at which 90% of isolates ratio (AUC/MIC90) were calculated to estimate the antibacterial effects in AMs. The TA of AZM in AMs following administration of aerosol formulation was markedly greater than that following administration of oral formulation. In addition, the AUC/MIC90 of AZM in AMs was markedly higher than the effective values. This indicates that the aerosol formulation could be useful for the treatment of respiratory infections caused by pathogenic microorganisms infected in AMs. This study suggests that aerosolized AZM is an effective pulmonary drug delivery system for the treatment of respiratory infections.


Turkish Version Study of "Parkinson's Disease Quality of Life Questionnaire" (PDQL).
This study aimed to evaluate Turkish transcultural adaptation, reliability, and validity of "Parkinson's Disease Quality of Life Questionnaire" (PDQL), which has been developed to assess the quality of life of patients with Parkinson's disease. Eighty-nine patients who were included in the study were diagnosed with Parkinson's disease. They were presented to the outpatient clinic of the Istanbul Faculty of Medicine, Physical Medicine and Rehabilitation department between July 2005 and April 2008. The Turkish translated version of PDQL (PDQL-TR), "Unified Parkinson's Disease Rating Scale" (UPDRS), and Hoehn & Yahr (HY) were used as the main outcome measures. The calculated Pearson correlation coefficient for PDQL, H&Y, and UPDRS ranged between -0.36 and -0.80. Cronbach's alpha coefficients of the PDQL-TR complete questionnaire and subdomains were higher than 0.80. The absolute value of item-total score correlation coefficients by Pearson fluctuated between 0.40 (question 4-total score) and 0.88 (question 26-total score), whereas the subdomain-total score correlation coefficients ranged between 0.81 and 0.96. PDQL-TR showed high stability according to the ICC results, ranging between 0.75 (Emotional Function) and 0.88 (Parkinsonian Symptoms). The psychometric features of PDQL-TR are satisfactory, and our results are consistent with the previous literature findings. PDQL-TR is a valid and reliable tool to measure health-related quality of life in patients with Parkinson's disease.


The complexity of implementation factors in professional pharmacy services.
Implementation of professional pharmacy services is a complex process, in which multi-level factors interact and influence implementation process and outcomes at various levels or domains. In this paper the terms that have been used to describe the factors that influence implementation of evidence-based services and their domains are presented. The complex 'cause-and-effect' interactions by which implementation factors appear to interact throughout the implementation process are also discussed. Identifying and understanding these complex and causal relationships between different implementation factors, represents a key process in the implementation of any service, in order to assist in the development of tailored implementation strategies. Future research should be directed to gain an understanding of the nature of individual implementation factors, their cause-and-effect interactions, and their relationships. Implementation strategies are less likely to succeed unless these are identified and targeted to the causes identified when designing and planning an implementation strategy.


A neural network technique for remeshing of bone microstructure.
Today, there is major interest within the biomedical community in developing accurate noninvasive means for the evaluation of bone microstructure and bone quality. Recent improvements in 3D imaging technology, among them development of micro-CT and micro-MRI scanners, allow in-vivo 3D high-resolution scanning and reconstruction of large specimens or even whole bone models. Thus, the tendency today is to evaluate bone features using 3D assessment techniques rather than traditional 2D methods. For this purpose, high-quality meshing methods are required. However, the 3D meshes produced from current commercial systems usually are of low quality with respect to analysis and rapid prototyping. 3D model reconstruction of bone is difficult due to the complexity of bone microstructure. The small bone features lead to a great deal of neighborhood ambiguity near each vertex. The relatively new neural network method for mesh reconstruction has the potential to create or remesh 3D models accurately and quickly. A neural network (NN), which resembles an artificial intelligence (AI) algorithm, is a set of interconnected neurons, where each neuron is capable of making an autonomous arithmetic calculation. Moreover, each neuron is affected by its surrounding neurons through the structure of the network. This paper proposes an extension of the growing neural gas (GNN) neural network technique for remeshing a triangular manifold mesh that represents bone microstructure. This method has the advantage of reconstructing the surface of a genus-n freeform object without a priori knowledge regarding the original object, its topology, or its shape.


Inferring space from time: On the relationship between demography and environmental suitability in the desert plant O. rastrera.
Demographic analyses and ecological niche modeling (ENM) are two popular tools that address species persistence in relation to environmental conditions. Classic demography provides detailed information about the mechanisms that allow a population to grow or remain stable at a local scale, while ENM infers distributions from conditions suitable for species persistence at geographic scales by relating species' occurrences with environmental variables. By integrating these two tools, we may better understand population processes that determine species persistence at a geographic scale. To test this idea, we developed a model that relates climate to demography of the cactus Opuntia rastrera using 15 years of data from one locality. Using this model we determined the geographic area where populations would have positive growth rates given its climatic conditions. The climate-dependent demographic model showed poor performance as a distribution model, but it was helpful in defining some mechanisms that determine species' distributions. For instance, high rainfall had a negative impact on the population growth rate by increasing mortality. Rainy areas to the west of the distribution of O. rastrera were identified as unsuitable both by our climate-dependent demographic model and by a popular ENM algorithm (MaxEnt), suggesting that distribution is constrained by excessive rains due to high mortality. Areas projected to be climatically suitable by MaxEnt were not related with higher population growth rates. Instead, we found a strong correlation between environmental distance to the niche centroid (center of the niche hypervolume, where optimal conditions may occur) and population growth rate, meaning that the niche centroid approach is helpful in finding high-fitness areas.


[Influence of nutrient medium composition on the production of capsule polysaccharide by Streptococcus pneumonia 19A serotype].
Evaluate accumulation of capsule polysaccharide by Streptococcus pneumoniae 19A strain in semisynthetic nutrient medium including various amino acid sources. Comparative evaluation of the production of capsule polysaccharide by the strain belonging to one of the most widespread S. pneumoniae serotype (19A) was performed by using rocket immunoelectrophoresis. The bacteria were cultivated in semisynthetic liquid nutrient media of varying composition. Among 4 sources of nitrogen (aminopeptide, acid and pancreatic hydrolysate of casein, soy peptone) added to salt nutrient medium supplemented with glucose and vitamins, casein and soy peptone were shown to promote the maximum synthesis of capsule polysaccharide independently of the cultivation time. Supplementation of the medium with sulfates of iron, zinc and manganese, as well as pH decrease to acid values significantly reduced the level of capsule polysaccharide in the culture liquid. The maximum growth of bacteria was observed at 11 hours after the start of cultivation in a 10 L volume fermenter in semisynthetic nutrient medium with soy peptone. Accumulation of capsule polysaccharide in the culture liquid continued to the end of the observation period (24 hours) and by the end of the process reached 193 mcg/ml. Further study of influence of vitamins, carbohydrates, CO2 concentration on the synthesis of high molecular capsule polysaccharide by bacteria belonging to various pneumococcus serotypes is reasonable.


Halvade-RNA: Parallel variant calling from transcriptomic data using MapReduce.
Given the current cost-effectiveness of next-generation sequencing, the amount of DNA-seq and RNA-seq data generated is ever increasing. One of the primary objectives of NGS experiments is calling genetic variants. While highly accurate, most variant calling pipelines are not optimized to run efficiently on large data sets. However, as variant calling in genomic data has become common practice, several methods have been proposed to reduce runtime for DNA-seq analysis through the use of parallel computing. Determining the effectively expressed variants from transcriptomics (RNA-seq) data has only recently become possible, and as such does not yet benefit from efficiently parallelized workflows. We introduce Halvade-RNA, a parallel, multi-node RNA-seq variant calling pipeline based on the GATK Best Practices recommendations. Halvade-RNA makes use of the MapReduce programming model to create and manage parallel data streams on which multiple instances of existing tools such as STAR and GATK operate concurrently. Whereas the single-threaded processing of a typical RNA-seq sample requires ∼28h, Halvade-RNA reduces this runtime to ∼2h using a small cluster with two 20-core machines. Even on a single, multi-core workstation, Halvade-RNA can significantly reduce runtime compared to using multi-threading, thus providing for a more cost-effective processing of RNA-seq data. Halvade-RNA is written in Java and uses the Hadoop MapReduce 2.0 API. It supports a wide range of distributions of Hadoop, including Cloudera and Amazon EMR.


A Probabilistic Digital Twin for Leak Localization in Water Distribution Networks Using Generative Deep Learning.
Localizing leakages in large water distribution systems is an important and ever-present problem. Due to the complexity originating from water pipeline networks, too few sensors, and noisy measurements, this is a highly challenging problem to solve. In this work, we present a methodology based on generative deep learning and Bayesian inference for leak localization with uncertainty quantification. A generative model, utilizing deep neural networks, serves as a probabilistic surrogate model that replaces the full equations, while at the same time also incorporating the uncertainty inherent in such models. By embedding this surrogate model into a Bayesian inference scheme, leaks are located by combining sensor observations with a model output approximating the true posterior distribution for possible leak locations. We show that our methodology enables producing fast, accurate, and trustworthy results. It showed a convincing performance on three problems with increasing complexity. For a simple test case, the Hanoi network, the average topological distance (ATD) between the predicted and true leak location ranged from 0.3 to 3 with a varying number of sensors and level of measurement noise. For two more complex test cases, the ATD ranged from 0.75 to 4 and from 1.5 to 10, respectively. Furthermore, accuracies upwards of 83%, 72%, and 42% were achieved for the three test cases, respectively. The computation times ranged from 0.1 to 13 s, depending on the size of the neural network employed. This work serves as an example of a digital twin for a sophisticated application of advanced mathematical and deep learning techniques in the area of leak detection.


The fate of bacteria in frozen red cells.
Units of blood were intentionally contaiminated with suspensions of either Aerobacter aerogenes, Escherichia coli, Bacillus subtilis, Enterobacter cloacae, Pseudomonas aeruginosa, Sarcina lutea, Seratia marcesens, Staphylococcus epidermidis, Streptococcus faecalis, Paracolabacterum aerogenoides (Enterobacter hafriae), Mima polymorpha or Acinetobacter- calcoaceticus. When inoculation was made prior to glycerolization, the subsequent glycerolization, freezing, thawing, and deglycerolization resulted in roughly a two log reduction in the number of bacteria. When inoculation was made with a final concentration of between 10(1) and 10(5) organisms per milliliter, immediately following deglycerolization or following washing without glycerolization and freezing, no increase in the number of bacteria was seen after 72 hours storage at 4 C. Three of the 12 organisms studied decreased in number during 72 hours of storage. These data suggest that the current 24-hour limit on the post-thaw storage of frozen red blood cells may be unnecessarily restrictive.


Slow sodium-dependent potential oscillations contribute to ectopic firing in mammalian demyelinated axons.
Ectopic action potentials can arise at regions of axonal demyelination, and are believed to contribute to a range of symptoms in patients with demyelinating conditions such as multiple sclerosis. The mechanism(s) by which the ectopic impulses are generated are uncertain. We have previously shown that such ectopic activity can result from inward potassium currents. Paradoxically, the potassium channel blocking agent 4-aminopyridine (4AP) can sometimes cause ectopic activity in demyelinating lesions. To study this phenomenon we have made intra-axonal recordings of ectopic activity in demyelinated axons, both in the presence and absence of 5 mM 4AP. 4AP promoted a pattern of firing which was observed, albeit less frequently, in demyelinated axons in the absence of this drug, namely trains of single impulses, or trains of short, high-frequency bursts of impulses. When recorded close to the demyelinated lesion, the impulses were generated by an underlying, almost sinusoidal oscillation of the membrane potential. This oscillation was abolished by the sodium channel blocking agent tetrodotoxin (0.1-1 microM). We conclude that the ectopic spiking induced by 4AP is generated by membrane potential oscillations associated with the site of demyelination. The sodium-dependent current underlying these oscillations, together with the prolonged inward potassium currents which we have previously described, may contribute to the generation of ectopic discharges in a range of disorders of myelinated axons.


Association of intronic repetition of SLC26A4 gene with Hashimoto thyroiditis disease.
Intronic microsatellites repeats were implicated in the pathogenic mechanisms of several diseases. SLC26A4 gene, involved in the genetic susceptibility of autoimmune thyroid disease (AITD), harbours large non-coding introns. Using the tandem repeat finder (TRF) Software, two new polymorphic microsatellite markers, rs59736472 and rs57250751, located at introns 10 and 20, respectively, were identified. A case-control design including 308 patients affected with AITD (146 GD, 90 HT and 72 PIM) and 212 unmatched healthy controls were performed for each marker (rs59736472, D7S2459 and rs57250751). Furthermore, we used PHASE 2.0 version to reconstruct haplotypes, Kolmogorov-Smirnov (KS) and the Clump analysis program for multivariate analysis. The fluorescent genotyping revealed three alleles (106,112 and 115 bp) for rs57250751 and 12 alleles for both D7S2459 and rs59736472 ranging from 134 to 156 bp and from 144 to 168 bp, respectively. The case-control analysis confirmed the positive association of D7S2459 with Hashimoto thyroiditis (HT) disease previously reported. Moreover, a significant association was found only with rs59736472 and HT disease. Haplotype-specific analysis showed that the 140-148-115 haplotype may increase the risk of HT (χ2=9.8, 1 df, P=0.0017, OR=2.07, IC [1.27-3.36]). Consequently, considering the number of repetitions of both D7S2459 and rs59736472, we found 15 alleles ranging from 45 to 59 repetitions. The case-control analysis showed a significant association of the 55 repetition with HT disease (χ2=6.32, 1 df, p c=0.012, OR=1.74, IC [1.1-2.76]). In conclusion, we suggest the association of longer alleles of intron 10 of SLC26A4 gene with HT disease.


[Effect of occupational hazard factor type of acetylenogen dust on occupational health risk assessment results].
To explore the differences of health hazards between acetylenogen dust as two different occupational hazards of "particles not otherwise regulated" and "calcium carbide". A calcium carbide production enterprise in northwest China was selected. The occupational health survey was used to obtain the spatial distribution and population distribution of calcium carbide dust. Occupational health testing was performed to determine the time-weighted average exposure concentration(C_(TWA)) and excursion limits of calcium carbide dust in the workplace and the main exposure positions. The Singapore semi-quantitative risk matrix was used for occupational risk assessment. The "Classification of Occupational Disease Hazards in the Workplace Part 1: Productive Dust"(GBZ/T 229.1-2010) was used for the operation classification. The calcium carbide dust was regarded as two different occupational hazards: "particles not otherwise regulated" and "calcium carbide", the differences of health hazards between which were compared. PC-TWA of calcium carbide was performed at 2.28 mg/m~3. The heat amount of CaC_2 and CaO released after reacting with water was compared by using Hess' s law, and the reaction rates of the two reactions are compared. The concentrations of calcium carbide dust in the workplace of belt inspection position and the calcium carbide crusher was higher(14.3-15.7 mg/m~3), and the concentrations of the driver in the calcium carbide loader was higher(13.8 mg/m~3). The result of the risk assessment show that the health risks of calcium carbide dust in various positions are basically low risk. Only when the type of damage was "calcium carbide", the risk of the driver of calcium carbide loader increase to medium. The operation classification result show that the hazard classification of calcium carbide dust in each post is basically relatively harmless. Only the type of damage was "calcium carbide", the classification of the driver of the calcium carbide crusher loader increased to moderately hazardous operations. As for the "calcium carbide", the operation classification index of calcium carbide were higher than that of "particles not otherwise regulated". CaC_2 reacting with water released more heat than that of equal amount CaO. In the current production process of calcium carbide dust, except for the crushing section, its occupational health risk was low, but the health risk level of calcium carbide as "calcium carbide" was higher, indicating that the evaluation of calcium carbide dust as "particles not otherwise regulated" may underestimate its occupational hazard.


Dual-specificity phosphatase 6 (DUSP6): a review of its molecular characteristics and clinical relevance in cancer.
Mitogen-activated protein kinases (MAPKs) are the main regulators of cellular proliferation, growth, and survival in physiological or pathological conditions. Aberrant MAPK signaling plays a pivotal role in carcinogenesis, which leads to development and progression of human cancer. Dual-specificity phosphatase 6 (DUSP6), a member of the MAPK phosphatase family, interacts with specifically targeted extracellular signal-regulated kinase 1/2 via negative feedback regulation in the MAPK pathway of mammalian cells. This phosphatase functions in a dual manner, pro-oncogenic or tumor-suppressive, depending on the type of cancer. To date, the tumor-suppressive role of DUSP6 has been demonstrated in pancreatic cancer, non-small cell lung cancer, esophageal squamous cell and nasopharyngeal carcinoma, and ovarian cancer. Its pro-oncogenic role has been observed in human glioblastoma, thyroid carcinoma, breast cancer, and acute myeloid carcinoma. Both roles of DUSP6 have been documented in malignant melanoma depending on the histological subtype of the cancer. Loss- or gain-of-function effects of DUSP6 in these cancers highlights the significance of this phosphatase in carcinogenesis. Development of methods that use the DUSP6 gene as a therapeutic target for cancer treatment or as a prognostic factor for diagnosis and evaluation of cancer treatment outcome has great potential. This review focuses on molecular characteristics of the DUSP6 gene and its role in cancers in the purview of development, progression, and cancer treatment outcome.


Relationship of hormone dependency to estrogen receptor and adenosine 3',5'-cyclic monophosphate-binding proteins in rat mammary tumors.
Estrogen-binding activity and the binding activity of adenosine 3',5'-cyclic monophosphate (cAMP) were assayed in the cytosol of biopsy specimens from 70 rat mammary tumors. The response of these tumors to host ovariectomy was followed. The hormone dependency of the tumors was closely related to the relative concentrations of estrogen receptor (ER) and cAMP-binding protein (CR) in the tumor cytosol. When ER and CR were expressed as the ER/CR ratio, 36 of 38 (95%) hormone-dependent tumors had ratios of 35 X 10(-3) or more, whereas 31 of 32 (97%) hormone-independent tumors had ratios of less than 35 X 10(-3). When ER alone was measured, hormone dependency could be correctly predicted in only 60% of the tumors because the ER range for individual tumors of both types greatly overlapped. Ratios of ER/CR were also elevated in normal estrogen target tissues as compared to those in normal estrogen nontarget tissues. Thus determination of the ER/CR ratio in tumor cytosols appeared to be a more reliable method of predicting hormone dependency than was determination of ER alone.


Influence of titanium on in vitro fibroblast-Porphyromonas gingivalis interaction in peri-implantitis.
Titanium wear particles have been found in peri-implant tissues, but their role in the pathogenesis of peri-implantitis remains unclear. We aimed to determine the in vitro inflammatory responses of peri-implant granulation tissue fibroblasts (PIGFs) to titanium particles alone and in the presence of viable Porphyromonas gingivalis. Peri-implant granulation tissue fibroblasts were challenged either with TiO2 particles, P. gingivalis or a combination of TiO2 particles and P. gingivalis. Gene expression and protein production of pro-inflammatory mediators by PIGFs were measured with PCR and ELISA, respectively. Higher doses of TiO2 were toxic to PIGFs and in sub-toxic doses, TiO2 caused an increase in gene expression of tumour necrosis factor (TNF)-A and increased protein production of TNF-α, interleukin (IL)-6 and IL-8. A challenge with P. gingivalis alone induced gene expression of TNF-A, IL-1β, IL-6 and IL-8. A combined challenge with TiO2 and P. gingivalis caused a stronger increase in gene expression of TNF-A and protein production of TNF-α and MCP-1 than P. gingivalis alone. TiO2 particles and P. gingivalis, individually, can induce pro-inflammatory responses in PIGFs. Furthermore, TiO2 particles and viable P. gingivalis further enhance gene expression and production of TNF-α by PIGFs. Therefore, Ti wear particles in the peri-implant tissues in combination with P. gingivalis infection may contribute to the pathogenesis of peri-implantitis by enhancing the inflammation in peri-implant tissues.


The Addition of ViriditecTM Aqueous Ozone to Peracetic Acid as an Antimicrobial Spray Increases Air Quality While Maintaining Salmonella Typhimurium, Non-pathogenic Escherichia coli, and Campylobacter jejuni Reduction on Whole Carcasses.
Currently, the most utilized antimicrobial in processing facilities is peracetic acid, PAA; however, this chemical is increasingly recognized as a hazard to human health. Preliminary evidence suggests that ozone, when introduced in a specific manner, can reduce the noxious nature of PAA. Therefore, the objective of the current study was to evaluate the efficacy of TetraClean Systems aqueous ozone, O3, in combination with PAA as an antimicrobial spray on whole chicken carcasses. This trial used 70 whole hen carcasses (7 treatments; 10 replications) that were inoculated in a 400 mL cocktail containing Salmonella, Escherichia coli, and Campylobacter (107 CFU/mL) and allowed to adhere for 60 min at 4°C for a final concentration of 105 to 106 CFU/g. The experimental 5 s (4×) spray treatments included: a no treatment negative control, TW; TW + O3 (10 ppm), TW + PAA (50 ppm), TW + PAA (500 ppm), TW + O3 + PAA (50 ppm), and TW + O3 + PAA (500 ppm). During treatment application, ambient PAA vapor was measured with a ChemDAQ Safecide PAA vapor sensor. After treatment, carcasses were immediately rinsed in 400 mL of nBPW for 2 min. Following rinsing, the dot method was utilizing for enumeration with 10 μL of rinsate being serially diluted, plated on XLD and mCCDA agar, and incubated aerobically at 37°C for 24 h or microaerophilically at 42°C for 48 h. Log-transformed counts were analyzed using ANOVA in JMP 14.0. Means were separated using Tukey's HSD when P ≤ 0.05. There was a significant treatment effect among Salmonella, E. coli, and Campylobacter counts, and a significant treatment effect among ambient PAA (P < 0.05). TW + O3 + PAA (500 ppm), reduced Salmonella significantly compared to TW (5.71 and 6.30 log CFU/g). Furthermore, TW + PAA (500 ppm), reduced the presence of E. coli significantly compared to TW or no treated control (5.57 and 6.18 log CFU/g). Also, TW + PAA (50 ppm), TW + PAA (500 ppm), and TW + O3 + PAA (500 ppm) significantly reduced Campylobacter compared to carcasses not treated (4.80, 4.81, and 4.86 log CFU/g). Lastly, the addition of ozone significantly reduced the ambient PAA when O3 was added to 500 ppm of PAA, as TW + O3 + PAA (500 ppm) produced less ambient PAA than TW + PAA (500 ppm) (0.052 and 0.565 ppm). In conclusion, the addition of ozone to PAA may demonstrated the ability to effectively reduce ambient PAA, thus increasing employee safety.


[A case of advanced gastric cancer with obstructive jaundice that responded to TS-1/CPT-11 combination therapy after percutaneous transhepatic cholangio drainage].
TS-1/CPT-11 combination therapy was carried out in a case of advanced gastric cancer with liver and lymph node metastases and obstructive jaundice after percutaneous transhepatic cholangio drainage (PTCD). Regression of the primary carcinoma and reduction in size of metastases were observed. Grade 1 fatigue and grade 2 neutropenia were noted as adverse reactions to the treatment. TS-1/CPT-11 combination therapy was useful in this case of advanced gastric cancer with liver and lymph node metastases.


Mechanistic study of beta-substituent effects on the mechanism of ketone reduction by SmI(2).
The rate constants for the reduction of 2-butanone, methylacetoacetate, N, N-dimethylacetoacetamide, and a series of 4'- and 2'-substituted acetophenone derivatives by SmI(2) were determined in dry THF using stopped-flow absorption decay experiments. Activation parameters for the electron-transfer processes in each series of compounds were determined by a temperature-dependence study over a range of 30 to 50 degrees C. Two types of reaction pathways are possible for these electron-transfer processes. One proceeds through coordination (Scheme 1) while the other involves chelation (Scheme 2). The results described herein unequivocally show that both coordination and chelation provide highly ordered transition states for the electron-transfer process but the presence of a chelation pathway dramatically increases the rate of the reduction of these substrates by SmI(2). The ability of various functional groups to promote a chelated reaction pathway plays a crucial role in determining the rate of the reaction. Among the 2'-substituted acetophenone series, the presence of a fluoro, amino, or methoxy substituent enhances the rate of reduction compared to the 4'-analogues. In particular, the presence of a 2'-fluoro substituent on acetophenone provides a highly ordered transition state and considerably enhances the rate of ketone reduction.


Inhibition of Microbiota-dependent Trimethylamine N-Oxide Production Ameliorates High Salt Diet-Induced Sympathetic Excitation and Hypertension in Rats by Attenuating Central Neuroinflammation and Oxidative Stress.
Excessive dietary salt intake induces neuroinflammation and oxidative stress in the brain, which lead to sympathetic excitation, contributing to hypertension. However, the underlying mechanisms remain elusive. Accumulating evidence reveals that trimethylamine-N-oxide (TMAO), a gut microbiota-derived metabolite, is implicated in the pathogenesis of multiple cardiovascular diseases. The present study sought to determine whether central TMAO is elevated and associated with neuroinflammation and oxidative stress in the brain after long-term high salt (HS) diet intake and, if so, whether inhibition of TMAO generation ameliorates HS-induced sympathetic excitation and hypertension. Sprague-Dawley rats were fed either a HS diet or a normal salt (NS) diet and simultaneously treated with vehicle (VEH) or 1.0% 3,3-Dimethyl-1-butanol (DMB, an inhibitor of trimethylamine formation) for 8 weeks. HS + VEH rats, compared with NS + VEH rats, had elevated TMAO in plasma and cerebrospinal fluid (CSF), increased blood pressure (BP), and increased sympathetic drive as indicated by the BP response to ganglionic blockade and plasma norepinephrine levels. HS-induced these changes were attenuated by DMB, which significantly reduced TMAO in plasma and CSF. Neuroinflammation as assessed by proinflammatory cytokine expression and NF-κB activity and microglial activity, and oxidative stress as measured by NAD(P)H oxidase subunit expression and NAD(P)H activity and reactive oxygen species (ROS) production in the hypothalamic paraventricular nucleus (PVN) were increased in HS + VEH rats but were decreased by DMB. DMB had no effects on above measured parameters in NS rats. The results suggest that long-term HS diet intake causes elevation in TMAO in the circulation and brain, which is associated with increased neuroinflammation and oxidative stress in the PVN, an important cardiovascular regulatory center. Inhibition of TMAO generation ameliorates HS-induced sympathetic excitation and hypertension by reducing neuroinflammation and oxidative stress in the PVN.


Genetic basis of sleep bruxism and sleep apnea-response to a medical puzzle.
Sleep bruxism (SB) and obstructive sleep apnea (OSA) are co-occurring sleep conditions. The study aimed to evaluate the association of selected single-nucleotide polymorphisms (SNPs) occurring within the genes of the serotonin and dopamine pathways in SB and OSA and investigate the relationship between them. The study group included 100 Caucasian patients. SB and OSA were diagnosed in 74 and 28 patients, respectively. In addition, 125 unrelated Caucasian healthy blood donors served as randomly selected controls to enable comparison of polymorphisms. The following SNPs were analyzed: rs2770304 and rs6313 within the serotonin receptor encoding gene (HTR2A), rs4680 polymorphism of the catechol-O-methyltransferase (COMT) gene, and rs686 within the dopamine receptor (DRD1) encoding gene. The prevalence of the DRD1 rs686 G variant (GG homozygosity) was found to be high in the study group compared to the control group. Bruxism episode index (BEI) was found to be significantly increased in the HTR2A rs6313 TT homozygotes compared to the heterozygous patients. Moreover, within a group of the HTR2A rs2770304 TT homozygous cases, a statistically significant correlation was observed between BEI and apnea-hypopnea index. These results indicate that DRD1 rs686 may potentially affect predisposition to SB, that HTR2A rs6313 SNP may be involved in SB pathogenesis, and that HTR2A rs2770304 polymorphism might contribute to the association between SB and OSA. This suggests a possible genetic contribution to the etiology of primary SB.


Formation of trans-activation competent HIV-1 Rev:RRE complexes requires the recruitment of multiple protein activation domains.
The HIV-1 Rev trans-activator is a nucleocytoplasmic shuttle protein that is essential for virus replication. Rev directly binds to unspliced and incompletely spliced viral RNA via the cis-acting Rev Response Element (RRE) sequence. Subsequently, Rev oligomerizes cooperatively and interacts with the cellular nuclear export receptor CRM1. In addition to mediating nuclear RNA export, Rev also affects the stability, translation and packaging of Rev-bound viral transcripts. Although it is established that Rev function requires the multimeric assembly of Rev molecules on the RRE, relatively little is known about how many Rev monomers are sufficient to form a trans-activation competent Rev:RRE complex, or which specific activity of Rev is affected by its oligomerization. We here analyzed by functional studies how homooligomer formation of Rev affects the trans-activation capacity of this essential HIV-1 regulatory protein. In a gain-of-function approach, we fused various heterologous dimerization domains to an otherwise oligomerization-defective Rev mutant and were able to demonstrate that oligomerization of Rev is not required per se for the nuclear export of this viral trans-activator. In contrast, however, the formation of Rev oligomers on the RRE is a precondition to trans-activation by directly affecting the nuclear export of Rev-regulated mRNA. Moreover, experimental evidence is provided showing that at least two protein activation domains are required for the formation of trans-activation competent Rev:RRE complexes. The presented data further refine the model of Rev trans-activation by directly demonstrating that Rev oligomerization on the RRE, thereby recruiting at least two protein activation domains, is required for nuclear export of unspliced and incompletely spliced viral RNA.


Inhibition of RNA synthesis in Neurospora crassa hyphae treated with picolinic acid.
Picolinic acid, a metal-chelating agent, blocks the nuclei of germinating conidia of Neurospora crassa in the G1 phase. The addition of picolinic acid to exponentially growing cultures of N. crassa in glucose medium causes an immediate inhibition of growth. Concentrations higher than 50 mM completely inhibit growth of mycelia, whereas lower concentrations (20-30 mM) give only a partial inhibition. Picolinic acid, added at the concentration of 20 mM, immediately blocks the accumulation of stable RNA, while protein accumulation continues at an unchanged rate for at least 30 min and only thereafter is slightly inhibited. DNA accumulation rapidly slows down after the addition of the drug. The level of polyadenylated RNA decreases quickly after the addition of picolinic acid, and the rate of its synthesis also appears to be inhibited. Picolinic acid greatly affects the metabolism of stable RNA in N. crassa: both processing and transcription of rRNA are sequentially inhibited with a moderate accumulation of precursor rRNA. It is concluded that the effects of picolinic acid on the nuclear division cycle in Neurospora may be related to the requirement of specific gene products to enter the S phase, whose synthesis is inhibited following the reduction of polyadenylated RNA synthesis.


Congenital hyperekplexia: five sporadic cases.
We report fives sporadic cases of hyperekplexia or startle disease characterized by a highly exaggerated startle reflex and tonic attacks. Affected neonates suffer from prolonged periods of stiffness and are at risk for sudden death from apnea. An early diagnosis is needed. Sudden loud sounds, unexpected tactile stimuli or percussion at the base of the nose can also elicit excessive jerking or tonic attack. The diagnosis of hyperekplexia is a purely clinical one. A defect of the alpha1 subunit of inhibitory glycine receptor (GLRA1) has been observed in the dominant form with a mutation in the chromosome 5. Clonazepam is effective and decreases the severity of the symptoms. The disease tends to improve after infancy and the psychomotor development is normal. The major form of "hyperekplexia" should be considered whenever one is confronted with neonatal hypertonicity associated with paroxysmal tonic manifestations (without electroencephalography anomalies). the diagnosis of hyperekplexia should be evaluated in any neonate with tonic attacks without evident cause.


Coordinated modifications in mesophyll conductance, photosynthetic potentials and leaf nitrogen contribute to explain the large variation in foliage net assimilation rates across Quercus ilex provenances.
Leaf dry mass per unit area (LMA) has been suggested to negatively affect the mesophyll conductance to CO2 (gm), the most limiting factor for photosynthesis per unit leaf area (AN) in many evergreens. Several anatomical traits (i.e., greater leaf thickness and thicker cell walls) constraining gm could explain the negative scaling of gm and AN with LMA across species. However, the Mediterranean sclerophyll Quercus ilex L. shows a major within-species variation in functional traits (greater LMA associated with higher nitrogen content and AN) that might contrast the worldwide trends. The objective of this study was to elucidate the existence of variations in other leaf anatomical parameters determining gm and/or biochemical traits improving the capacity of carboxylation (Vc,max) that could modulate the relationship of AN with LMA across this species. The results revealed that gm was the most limiting factor for AN in all the studied Q. ilex provenances from Spain and Italy. The within-species differences in gm can be partly attributed to the variation in several leaf anatomical traits, mainly cell-wall thickness (Tcw), chloroplast thickness (Tchl) and chloroplast exposed surface area facing intercellular air spaces (Sc/S). A positive scaling of gm and AN with Vc,max was also found, associated with an increased nitrogen content per area. A strong correlation of maximum photosynthetic electron transport (Jmax) with AN further indicated a coordination between the carboxylase activity and the electron transport chain. In conclusion, we have confirmed the strong ecotypic variation in the photosynthetic performance of individual provenances of Q. ilex. Thus, the within-species increases found in AN for Q. ilex with increasing foliage robustness can be explained by a synergistic effect among anatomical (at the subcellular and cellular level) and biochemical traits, which markedly improved gm and Vc,max.


Stimulation of cytoplasmic DNA sensing pathways in vitro and in vivo.
In order to efficiently stimulate an innate immune response, DNA must be of sufficient length and purity. We present a method where double stranded DNA (dsDNA) which has the requisite characteristics to stimulate the cytoplasmic DNA sensing pathways can be generated cheaply and with ease. By the concatemerization of short, synthetic oligonucleotides (which lack CpG motifs), dsDNA can be generated to be of sufficient length to activate the cytosolic DNA sensing pathway. This protocol involves blunt end ligation of the oligonucleotides in the presence of polyethylene glycol (PEG), which provides an environment for efficient ligation to occur. The dsDNA concatemers can be used, following purification by phenol/chloroform extraction, to simulate the innate immune response in vitro by standard transfection protocols. This DNA can also be used to stimulate innate immunity in vivo by intradermal injection into the ear pinna of a mouse, for example. By standardizing the concatemerization process and the subsequent stimulation protocols, a reliable and reproducible activation of the innate immune system can be produced.