[Changes in expression of autophagy-related factors during acute contusion repair of skeletal muscle].
To explore the possible biological mechanism of skeletal muscle contusion repair through researching the changes in expression of autophagy-related genes and proteins in SD rats with acute skeletal muscle contusion. Six rats were randomly selected as the control group from 30 male SD rats, acute skeletal muscle contusion model were established in the remaining 24 rats with self-made hitter, then the model rats were randomly divided into 4 groups (3 d, 5 d, 7 d, 14 d groups, n=6). On the 3rd, 5th, 7th and 14th day after injury, injured gastrocnemius of each group was harvested. The morphological and the ultra-microstructure changes of gastrocnemius after injury were observed by HE staining and transmission electron microscope (TEM) respectively. The relative protein levels of (LC3-Ⅱ) and P62 of each group were observed by Western blot. The relative mRNA levels of atg7, atg10, atg12, atg16L1 of each group were observed by RTPCR. The results of HE staining showed that compared with the control group, the inflammation reached its peak on the 5th day after injury, new muscle fibers were clearly observed in 7 d group. The results of TEM showed that, compared with the control group, oncotic mitochondria could be clearly seen in the 3 d, 5 d, 7 d groups. Also, the Z line changed from disappearing to drift thickening, sarcoplasmic reticulum dilatation gradually improved, there was no evident difference between the 14 d group and the control group, suggesting that the damage has preliminarily healed. The results of Western blot showed that the expressions of LC3-Ⅱand P62 were increased at first and then decreased. The expression of LC3-Ⅱwas markedly up-regulated in the 3 d, 5 d, 7 d groups compared with the control group and the 14 d group (P<0.01). Similarly, compared with the control group, the expression of P62 reached its peak on the 3rd day after injury (P<0. 01), and returned to normal level on the 14th day. The results of RT-PCR showed that the expression of atg10 mRNA in the natural recovery group of 3 d, 5 d, 7 d, 14 d was firstly decreased and then increased, the atg10 mRNA was markedly down-regulated in the 3 d, 5 d, 7 d groups compared with the control group and the 14 d group (P<0. 01). The expression of atg7, atg12, atg16L1 mRNA was generally increased at first and then decreased, it was markedly up-regulated in the 3 d, 5 d, 7 d groups compared with the control group and the 14 d group (P<0.01, P<0.05, P<0.01). The above results indicate that the autophagy is involved in repair of skeletal muscle injury by its autophagyrelated factors,regularly changes after contusion, and the rate of damage repair may be related to the level of autophagy.


Entitlements, coping mechanisms and indicators of access to food: wollo region, ethiopia, 1987-88.
Vulnerability to famine and traditional responses to food insecurity in Wollo Region, Ethiopia are described. The timeliness of anthropometric and socio-economic indicators of access to food is then assessed, using data collected in Wollo by Save the Children Fund during 1987-88, a period of drought and subsequent food insecurity. The movements of different indicators are then examined for evidence of correspondence at sub-district level. The author concludes that although anthropometric status does not respond as early as crop yield or grain price, a deterioration in anthropometric status is detectable at a stage when livestock and migration indicators show little or no change and mortality rate remains unaffected. At sub-district level, changes in different indicators are not well-correlated.


[Fatal accidents and non-fatal injuries amongst seamen in Iceland 2001-2005].
This study aims at getting a comprehensive view of the incidence, nature and circumstances leading up to injuries in Icelandic waters in 2001-2005. The NOMESCO database at Landspitali University Hospital registers prospectively accidents at sea treated in the Emergency Department, including circumstances leading to the accident, vessel type, experience, task being performed, weather conditions etc. The Icelandic Marine Accident Investigation Board provided data on fatal injuries, and additional information on injuries at sea was collected from the Social Insurance Administration (SIA). Of 17 fatal accidents occurring in 2001-2005 14 were work related which amounts to 54/100.000 seamen/year. The SIA received 1787 injury reports (7% of registered seamen), 826 sought assistance at Landspitali, 52 were admitted with an average injury severity score of 5.5 (1-16) and no ensuing fatalities. Most accidents occurred on fishing vessels (87%), 51% thereof on trawlers. Experienced fishermen are most commonly injured, working on deck in daylight and stable weather. Fatalities have declined steadily in the last two decades, yet 7% of registered fishermen sustain injuries each year. These occur amongst experienced fishermen in good external conditions, which calls for revision of safety procedures on board.


Correlates of fractures in elderly, diabetic outpatients.
It is widely recognized that patients presenting diabetes are at increased risk for fractures. In a retrospective case-control study, 101 cases were selected from medical charts of outpatients older than 70 with diabetes mellitus and a fracture within the past 5 years. Glycosylated hemoglobin (HbA1c) had been measured within 4 months around the assessment. Each case was matched for sex and age with one control, diabetic patient with no fracture. HbA1c level was similar in both groups. Patients with fractures presented significantly lower BMIs than controls, and had a higher rate of declared osteoporosis and comorbidity. A small number of cases were using vitamin D supplements while more were treated with benzodiazepine, opiates and Selective serotonin reuptake inhibitors (SSRI). This study suggests that, rather than the tight control of blood glucose, other factors such as medication and comorbidity could be associated with fracture risk in elderly diabetics.


Assessing and increasing patient panel size in the public sector.
Panel management is a central component of the primary care medical home, but faces numerous challenges in the safety net setting. In the San Francisco Department of Public Health, many of our community-based primary care clinics have difficulty accommodating all patients seeking care. We evaluated patient panel size in our 7 clinics providing cradle-to-grave primary care services to more than 25,000 active patients. We adjusted panel size for age, gender, diagnoses, homelessness, and substance abuse; set related policies; and assessed the effects on our clinics. On the basis of our previous data and targets set by other safety net providers, we established a minimum of 1125 patients per full-time paid primary care provider (ie, full-time equivalent [FTE]) in April 2009. We calculated the target panel size each clinic would have if all their providers reached the minimum panel size goal and compared it with the panel size attained by the clinic. Nine months after establishing panel size policy, providers reached 82% of the aggregate target panel size. Five of the 7 clinics increased their adjusted panel size per FTE in the range of 2% to 23%. Two data-oriented and innovative clinics with some of the highest adjusted panel sizes per FTE largely maintained their panel size. Two clinics that had the lowest adjusted panel size per FTE realized a 23% and 8% respective gain; both clinics reduced barriers to new patient appointments. Two clinics acquired new providers and experienced a concomitant drop in panel size per FTE while the new clinicians expanded their panels. One of these clinics had difficulty managing high no-show rates and creating effective appointment templates. Routine data generation, review of data with administrators and providers, data-driven policies and panel size standards, and interventions to bolster team-based care are important tools for increasing capacity at our primary care clinics.


Perception of labour pain among the Yoruba ethnic group in Nigeria.
In Nigeria, it is generally assumed that labour is well tolerated and pain relief is not usually considered an important part of intra-partum care. This prospective study was carried out to assess mothers' perception of labour pain and determine any factor that may influence it. During the period of study, 281 women who delivered at Wesley Guild Hospital Ilesa, Nigeria were interviewed within 2 h of delivery to assess the severity of labour pain and desire for analgesia. Perception of labour pain was assessed using a 3-point verbal rating. The majority (68.3%) of women described labour pain as severe with only 5.3% describing it as mild. More than 86% of the women would want the pain relieved. Perception of pain was not influenced by age, parity and educational level. Management of pain in labour should form an important part of intra-partum care as is the case in developed countries.


Field desorption mass spectrometry in structural studies of polyene macrolide antibiotics: isolation and early identification of a pentaene macrolide antibiotic.
Isolation of a pentaene macrolide antibiotic (NSC-277813) from Streptomyces griseus (FCRC-21) fermentation broth is described. Using field desorption mass spectrometry and high resolution field desorption mass spectrometry on the intact and derivatized antibiotic and degradation products, the antibiotic was identified as fungichromin. The application of field desorption mass spectrometry in the identification of polyene antibiotics is discussed.


Chemoenzymatic synthesis of pyrrolo[2,1-b]quinazolinones: lipase-catalyzed resolution of vasicinone.
A facile synthesis of bronchodilatory pyrrolo[2,1-b]quinazoline alkaloids by azidoreductive cyclization strategy employing TMSCl-NaI and bakers' yeast is described. Both the chemical and enzymatic methods are mild and take place at room temperature in good yields. Further, synthesis and resolution of vasicinone has been carried out by employing different lipases. It has been observed that lipase PS provides acetate of (S)-vasicinone in 98% ee.


Association of ADAM33 gene polymorphisms with allergic asthma.
Asthma results from the interaction between genetic and environmental factors. ADAM33 gene on chromosome 20p13 is associated with asthma and airway hyperresponsiveness. This is a case-control study, where four SNPs S1 (rs3918396), T1 (rs2280091), T2 (rs2280090), V4 (rs2787094) of ADAM33 gene have been assessed in patients with allergic asthma and normal controls (95 patients and 86 normal). Blood samples of these participants have been genotyped by PCR and the RFLP method. There was no association between asthmatic patients and polymorphisms of alleles, genotypes and haplotypes of the ADAM33 gene. When categorizing the asthmatic patients in severe, moderate and mild groups, associations in the subcategories of asthmatic patients were found. There were associations between polymorphisms of C allele of T1 SNP with severe asthmatic patients and G allele of V4 SNP with moderate asthmatics respectively (P=0.006, P=0.01). There was a significant association between sensitivity to mite and polymorphism of C allele of T1 SNP (P=0.02). Besides, there was a significant association between sensitivity to weeds and genotype GG of V4 SNP (P=0.05). Polymorphisms of ADAM33 gene might be associated with severe asthma and sensitivity to aeroallergens in northeast of Iran, but further studies are needed to determine the polymorphisms in this area and other regions of our country.


WWP1: a versatile ubiquitin E3 ligase in signaling and diseases.
WW domain-containing E3 ubiquitin protein ligase 1 (WWP1) is a multifunction protein containing an N-terminal C2 domain, four tandem WW domains for substrate binding, and a C-terminal catalytic HECT domain for ubiquitin transferring. WWP1 has been suggested to function as the E3 ligase for several PY motif-containing proteins, such as Smad2, KLF5, p63, ErbB4/HER4, RUNX2, JunB, RNF11, SPG20, and Gag, as well as several non-PY motif containing proteins, such as TβR1, Smad4, KLF2, and EPS15. WWP1 regulates a variety of cellular biological processes including protein trafficking and degradation, signaling, transcription, and viral budding. WWP1 has been implicated in several diseases, such as cancers, infectious diseases, neurological diseases, and aging. In this review article, we extensively summarize the current knowledge of WWP1 with special emphasis on the roles and action of mechanism of WWP1 in signaling and human diseases.


Diphenylparabanic acid as a synthon for the synthesis of α-diketones and α-ketocarboxylic acids.
Diphenylparabanic acid was found to react with >2 equiv of organolithiums at -78 °C to effectively give the corresponding symmetrical α-diketones. However, upon treatment with 1 equiv of organolithium, the parabanic acid gave mainly 5-substituted 5-hydroxyimidazolidine-2,4-diones. On the other hand, Grignard reagents were less reactive toward the parabanic acid at low temperature, and selectively gave the corresponding 5-hydroxyimidazolidine-2,4-diones even if more than 1 equiv of the reagents was used. A tandem process in which the parabanic acid was first reacted with a Grignard reagent and then reacted in one-pot with an organolithium effectively gave the unsymmetrical α-diketone. 5-Substituted 5-hydroxyimidazolidine-2,4-diones were useful as versatile precursors for preparing α-ketocarboxylic acids as well as unsymmetrical α-diketones.


MyD88 as a therapeutic target for inflammatory lung diseases.
Myeloid differentiation primary response protein 88 (MyD88) is a critical adaptor protein involved in Toll-like and IL-1 receptor family signaling controlling innate immune responses and inflammation. Genetic deletion of MyD88 function results in profound suppression of inflammation and reduced resistance of the host to pathogens indicating non-redundant roles of MyD88. The TIR domain is critical for MyD88 dimerization and signaling for TLR and IL-1R family receptor. Areas covered: Emerging evidence suggests that chemical disruption of the TIR domain attenuates cell activation and inhibits in vivo MyD88-dependent inflammation. We review the development of MyD88 dimerization disruptors as a novel therapeutic approach of respiratory diseases with a focus on COPD. Expert opinion: There is a proof of concept that therapeutic targeting of MyD88 is feasible and first preclinical data are highly promising. This opens a great opportunity to treat exacerbations of COPD and other chronic respiratory diseases. However, extensive preclinical investigations and risk analyses are required with carefully evaluation of reduced host resistance and opportunistic infections.


[Morphology of the placenta in relation to glycemia status in pregnancy in diabetes mellitus].
Morphological investigations in 569 placentas of diabetic women have been performed. The histological results of 300 placentas from diabetic women delivered between 1969 and 1973 were compared to 269 placentas of diabetic women from the time period 1979 to 1983. The latest time period was characterised by normoglycemic metabolic control throughout pregnancy. Besides the morphological findings clinical results especially the perinatal mortality and morbidity during the two time periods are discussed. Morphological changes in the placenta of diabetic women are depending one the quality of metabolic control during pregnancy, independing on the White classes. Normoglycemia reduces placental maturation disorders, perinatal mortality and morbidity.


Characterization of Anthocyanins in Perilla frutescens var. acuta Extract by Advanced UPLC-ESI-IT-TOF-MSn Method and Their Anticancer Bioactivity.
The anthocyanin extract from a domestic Perilla cultivar (Perilla frutescens var. acuta) were isolated and characterized with high mass accuracy and multi-dimensional fragmentation by means of ultra-performance liquid chromatography (UPLC) and electrospray ionization-ion trap-time of flight mass spectrometry analysis (ESI-IT-TOF-MSn). The new developed and applied LC-MS method focused on in-depth screening of anthocyanin compounds with similar structures which also provided a new approach of anthocyanin characterization without the use of external standards. Selective detection of interested anthocyanins was achieved utilizing extracted ion chromatogram (EIC) analysis, while MSn spectra were recorded to allow identification of the anthocyanin based on characteristic fragmentation patterns. Seven anthocyanins including one feruloyl (Cyanidin 3-O-feruloylglucoside-5-O-glucoside), two caffeoyl (Cyanidin 3-O-caffeoylglucoside-5-O-glucoside, Cyanidin 3-O-caffeoylglucoside-5-O-malonylglucoside) and four coumaroyl substituted anthocyanins (Cis-shisonin, Malonyl-cis-shisonin, Shisonin, and Malonyl-shisonin) were identified. Annexin-V FITC/PI flow cytometric assay was performed to analyze the influence of anthocyanin extract of P. frutescens var. acuta on cell apoptosis. The results suggested that Perilla anthocyanins can induce Hela cell apoptosis by a dose dependent manner.


Publication games: In the web of reciprocity.
The present processes of research assessment, i.e. focusing on one or a few, related, scientometrics, foster questionable authorship practices, like gifting authorship to non-contributing people. An especially harmful one of these unethical practices is the formation of publication cartels, where authors offer gift authorship to each other reciprocally. Here, by developing a simple model and a simulation of the publication process I investigate how beneficial cartels can be and what measure can be used to restrict them. My results indicate that publication cartels can significantly boost members' productivity even if paper counts are weighted by the inverse of author number (the 1/n rule). Nevertheless, applying the 1/n rule generates conflicts of interest both among cartel members themselves and between cartel members and non-members which might lead to the self-purification of the academic publishing industry.


[Primary pleural lymphoma: a rare complication of tuberculosis pleural sequelae].
We report two cases of malignant lymphoma of B phenotype occurring after therapeutic pneumothorax for tuberculosis. In both cases, outcome was fatal without time for specific treatment. Mainly reported in Japan, this pathology seems to be less frequent in western countries. As for B phenotype lymphoma associated with immunodeficiency, association with Epstein Barr virus is reported. Definite diagnosis is difficult and requires surgical biopsy. Prognosis remains poor with a survival ranging from 3 to 6 month.


Constitutive equations for the flow behavior of entangled polymeric systems: application to star polymers.
A semimicroscopic derivation is presented of equations of motion for the density and the flow velocity of concentrated systems of entangled polymers. The essential ingredient is the transient force that results from perturbations of overlapping polymers due to flow. A Smoluchowski equation is derived that includes these transient forces. From this, an equation of motion for the polymer number density is obtained, in which body forces couple the evolution of the polymer density to the local velocity field. Using a semimicroscopic Ansatz for the dynamics of the number of entanglements between overlapping polymers, and for the perturbations of the pair-correlation function due to flow, body forces are calculated for nonuniform systems where the density as well as the shear rate varies with position. Explicit expressions are derived for the shear viscosity and normal forces, as well as for nonlocal contributions to the body force, such as the shear-curvature viscosity. A contribution to the equation of motion for the density is found that describes mass transport due to spatial variation of the shear rate. The two coupled equations of motion for the density and flow velocity predict flow instabilities that will be discussed in more detail in a forthcoming publication.


Application and comparison of high-speed countercurrent chromatography and high performance liquid chromatography in preparative enantioseparation of α-substitution mandelic acids.
Preparative enantioseparations of α-cyclopentylmandelic acid and α-methylmandelic acid by high-speed countercurrent chromatography (HSCCC) and high performance liquid chromatography (HPLC) were compared using hydroxypropy-β-cyclodextrin (HP-β-CD) and sulfobutyl ether-β-cyclodextrin (SBE-β-CD) as the chiral mobile phase additives. In preparative HPLC the enantioseparation was achieved on the ODS C18 reverse phase column with the mobile phase composed of a mixture of acetonitrile and 0.10 mol L-1 phosphate buffer at pH 2.68 containing 20 mmol L-1 HP-β-CD for α-cyclopentylmandelic acid and 20 mmol L-1 SBE-β-CD for α-methylmandelic acid. The maximum sample size for α-cyclopentylmandelic acid and α-methylmandelic acid was only about 10 mg and 5 mg, respectively. In preparative HSCCC the enantioseparations of these two racemates were performed with the two-phase solvent system composed of n-hexane-methyl tert.-butyl ether-0.1 molL-1 phosphate buffer solution at pH 2.67 containing 0.1 mol L-1 HP-β-CD for α-cyclopentylmandelic acid (8.5:1.5:10, v/v/v) and 0.1 mol L-1 SBE-β-CD for α-methylmandelic acid (3:7:10, v/v/v). Under the optimum separation conditions, total 250 mg of racemic α-cyclopentylmandelic acid could be completely enantioseparated by HSCCC with HP-β-CD as a chiral mobile phase additive in a single run, yielding 105-110 mg of enantiomers with 95-98% purity and 85-90% recovery. But, no complete enantioseparation of α-methylmandelic acid was achieved by preparative HSCCC with either of the chiral selectors due to their limited enantioselectivity. In this paper preparative enantioseparation by HSCCC and HPLC was compared from various aspects.


Epidemiology of acquired brain injury (ABI) in adults: prevalence of long-term disabilities and the resulting needs for ongoing care in the region of Flanders, Belgium.
In order to plan services for long-term care of those with acquired brain injury (ABI), the prevalence of ABI was investigated by using a postal inquiry sent to a representative sample of general practitioners, of whom 121 cooperated and registered 186 cases. The corresponding estimated prevalence of adults with ABI-related disabilities is 183 per 100 000 population. The large majority remained at home without professional assistance. One in five had resumed education or regained full employment, but over half spent their day at home with no specific activity. The need for specific care at home was present in almost half of the patients. One-quarter had employment-related needs, and one in five had living needs for specialized accommodation or supported living. In conclusion, this study primarily revealed unfulfilled needs in the area of day services aiming at a maximal social and professional reintegration.


Isolation and characterization of cDNA clones corresponding to two different human apoC-III alleles.
We have recently reported that the human apolipoprotein A-I (apoA-I) and apolipoprotein C-III (apoC-III) genes are physically linked and that the presence of a DNA insertion in the apoA-I gene is correlated with apoA-I-apoC-III deficiency in patients with premature atherosclerosis. In addition, the presence of a polymorphic restriction endonuclease site (SacI) in the 3' noncoding region of apoC-III mRNA has been correlated with hypertriglyceridemia in humans. In this study, we report the isolation and characterization of cDNA clones containing the entire apoC-III mRNA coding sequence. The nucleotide-derived apoC-III amino acid sequence indicates that the apoC-III primary translational product contains a 20 amino acid N-terminal extension, which conforms with the general properties of known signal peptides, and is highly homologous to the recently reported rat apoC-III signal peptide. The DNA-derived apoC-III amino acid sequence differs from the previously reported apoC-III amino acid sequence at four amino acid residues. More specifically, at positions +32, +33, +37, +39, the DNA sequence predicts Glu, Ser, Gln, Ala, respectively, while the previously reported sequence specifies Ser, Gln, Ala, Gln, respectively. Finally, isolation and characterization of apoC-III cDNA clones, with or without the polymorphic SacI restriction site, indicated that the apoC-III nucleotide sequence corresponding to the Sac+ and Sac- clones differs at three nucleotide sites; however, the amino acid sequence specified by the Sac+ and Sac- alleles is identical.


Withdrawal-induced antihypertensive effect of vasopressin in Doca-salt hypertension.
Arterial pressure and heart rate were recorded in three groups of conscious unrestrained rats before, during and after a three-hour intravenous infusion of arginine vasopressin (AVP; 20 ng/kg/min). Before infusion of AVP, basal arterial pressure was 152 +/- 3.2 mm Hg for the Doca-salt hypertensive group, 117 +/- 3.2 mm Hg for the uninephrectomized control group, and 111 +/- 2.1 mm Hg for the sham-operated control group. The changes in arterial pressure at each of the time points during the three-hour infusion of AVP were not significantly different among the three groups. Cessation of the AVP infusion was associated with a dramatic and prolonged fall of arterial pressure below preinfusion basal levels in the Doca-salt hypertensive group: pressure was 58 +/- 5.0 mm Hg below basal levels within 60 minutes of stopping the infusion and was still 45 +/- 5.6 mm Hg below four hours later. Pressure in this group was 107 +/- 5.7 mm Hg five hours after stopping the infusion. In contrast, the fall in pressure in the two control groups was small. Five hours after stopping the infusion, pressure was 108 +/- 2.3 mm Hg in the uninephrectomized group and 102 +/- 1.7 mm Hg in the sham-operated group. Thus, five hours after stopping the infusion, blood pressures among the three groups were not significantly different. The Doca-salt hypertensive group had been rendered normotensive.(ABSTRACT TRUNCATED AT 250 WORDS)


Prioritization of chemicals according to the degree of hazard in the aquatic environment.
Chemicals designated as "priority pollutants" or "toxics" have received special attention recently because the discharge of these compounds into public water is to be restricted to the maximum possible with little regard to water quality or economics. The selection of many of the 129 priority cemicals was not based on an objective scientific assessment of the exposure and effect data. In fact, for some compounds, including acenaphthene and 4-chlorophenyl-phenyl ether, the necessary data for listing were non-existent. As an alternative to arbitrarily listing or delisting chemicals for the purpose of prioity control, this paper suggests a promising scientific approach to selecting priority chemicals based on the principles of hazard assessment for chemicals in the aquatic environment. According o the hypothesis, the highest priority chemicals are those with the least margin of safety, defined as the gap between the no-observable-effect concentrations and the ambient exposure concentrations. The no-observable-effect concenrations are based on the results of chronic or sensitive life stage tests with aquatic organisms and the acceptable daily intake rate for fish eates. The ambient exposure concentrations are levels either measured in fish and water, or roughly estimated from a simple nomogram that requires only two of the following three factors: environmental release rate, ratio of dissipation to bioconcentration potential, or ambient residues in fish. THE CHEMICALS STUDIED TO ILLUSTRATE THIS APPROACH TO PRIORITIZING CHEMICALS BASED ON HAZARD ASSESSMENT ARE: polychlorinated biphenyls, di-2-ethylhexyl phthalate, linear alkylbenzene sulfonate, and pentachlorophenol.


Dietary fatty acid supplementation alters stress reactivity and performance in man.
Certain dietary polyunsaturated fatty acids, gamma linolenic (18:3n-6) and eicosapentaenoic (20:5n-3) acid, attenuate cardiovascular reactivity to stress in rats. To study their effects on cardiovascular reactivity to acute stress in man, 30 men were randomly assigned to one of three groups and given 28 day supplements of borage oil (containing 18:3n-6), fish oil (containing 20:5n-3), or olive oil (placebo). Reactivity to the Stroop colour-word conflict test was assessed prior to and following treatment. Borage oil alone attenuated blood pressure and heart rate responses to stress, increased skin temperature, and improved task performance. These data suggest that diet may be used to alter stress reactivity in man.


The Difference of Volatile Compounds in Female and Male Buds of Trichosanthes anguina L. Based on HS-SPME-GC-MS and Multivariate Statistical Analysis.
Trichosanthes anguina L. (family Cucurbitaceae) is a monoecious and diclinous plant that can be consumed as a vegetable and has anti-inflammatory and antioxidant effects. The chemical composition and content of volatile compounds in female and male buds of T. anguina were explored by headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) technology combined with multivariate statistical analysis. The results showed that the content of the volatile compounds was different between female and male buds. 2,2,6-trimethyl-6-vinyltetrahydro-2H-pyran-3-ol and 2,2,6-trimethyl-6-vinyldihydro-2H-pyran-3(4H)-one were the main volatile compounds in both female and male buds. Based on the multivariate statistical analysis of orthogonal projections to latent structures discriminant analysis (OPLS-DA) and t-test, the content of seven compounds was significantly different between female and male buds. The content of three compounds in male buds was higher than that in female, i.e., (E)-4,8-dimethyl-1,3,7-nonatriene, 1,5,9,9-tetramethyl-1,4,7-cycloundecatriene, and (E)-caryophyllene. Conversely, the content of (Z)-4-hexen-1-ol, (Z)-3-hexenyl benzoate, (Z)-3-hexenyl salicylate, and 2-hexen-1-ol in female buds was higher than that in male buds. This is the first report on the difference in the volatile compounds between female and male buds of T. anguina, which enriches the basic research on the monoecious and diclinous plant and provides a reference for the study of plant sex differentiation.


Amygdala norepinephrine levels after training predict inhibitory avoidance retention performance in rats.
Previous findings indicate that footshock and several drugs that modulate memory consolidation alter norepinephrine (noradrenaline) release in the amygdala, as assessed by in vivo microdialysis and high-performance liquid chromatography. Such findings suggest that norepinephrine release in the amygdala may be critical for regulating memory consolidation. The present study was the first to examine the relationship between norepinephrine release in the amygdala assessed after inhibitory avoidance training and 24-h retention performance within individual animals. Norepinephrine levels increased to > 300% of pretraining baseline 30 min after training and remained elevated for 2 h. In individual rats, the increase in norepinephrine levels after training correlated highly with 24-h retention performance. These findings indicate that the degree of activation of the noradrenergic system within the amygdala in response to a novel, emotionally arousing experience predicts the extent of long-term memory for that experience.


Analysis of multifrequency langevin composite ultrasonic transducers.
The multimode coupled vibration of Langevin composite ultrasonic transducers with conical metal mass of large cross-section is analyzed. The coupled resonance and anti-resonance frequency equations are derived and the effective electromechanical coupling coefficient is analyzed. The effect of the geometrical dimensions on the resonance frequency, the anti-resonance frequency, and the effective electromechanical coupling coefficient is studied. It is illustrated that when the radial dimension is large compared with the longitudinal dimension, the vibration of the Langevin transducer becomes a multifrequency multimode coupled vibration. Numerical methods are used to simulate the coupled vibration; the simulated results are in good agreement with those from the analytical results. Some Langevin transducers of large cross-section are designed and manufactured and their resonance frequencies are measured. It can be seen that the resonance frequencies obtained from the coupled resonance frequency equations are in good agreement with the measured results. It is expected that by properly choosing the dimensions, multifrequency Langevin transducers can be designed and used in ultrasonic cleaning, ultrasonic sonochemistry, and other applications.


Functional Analysis of Autophagy-Related Gene ATG12 in Potato Dry Rot Fungus Fusarium oxysporum.
Autophagy is an intracellular process in all eukaryotes which is responsible for the degradation of cytoplasmic constituents, recycling of organelles, and recycling of proteins. It is an important cellular process responsible for the effective virulence of several pathogenic plant fungal strains, having critical impacts on important crop plants including potatoes. However, the detailed physiological mechanisms of autophagy involved in the infection biology of soil-borne pathogens in the potato crop needs to be investigated further. In this study, the autophagy-related gene, FoATG12, in potato dry rot fungus Fusarium oxysporum was investigated by means of target gene replacement and overexpression. The deletion mutant ∆FoATG12 showed reduction in conidial formation and exhibited impaired aerial hyphae. The FoATG12 affected the expression of genes involved in pathogenicity and vegetative growth, as well as on morphology features of the colony under stressors. It was found that the disease symptoms were delayed upon being inoculated by the deletion mutant of FoATG12 compared to the wild-type (WT) and overexpression (OE), while the deletion mutant showed the disease symptoms on tomato plants. The results confirmed the significant role of the autophagy-related ATG12 gene in the production of aerial hyphae and the effective virulence of F. oxysporum in the potato crop. The current findings provid an enhanced gene-level understanding of the autophagy-related virulence of F. oxysporum, which could be helpful in pathogen control research and could have vital impacts on the potato crop.


Activation of the cAMP/CREB/inducible cAMP early repressor pathway suppresses andrographolide-induced gene expression of the pi class of glutathione S-transferase in rat primary hepatocytes.
Andrographolide (Ap) is a bioactive compound in Andrographis paniculata that is a Chinese herb. The pi class of glutathione S-transferase (GSTP) is one kind of phase II detoxification enzyme. Here we show that induction of GSTP protein and mRNA expression in rat primary hepatocytes by Ap was inhibited by forskolin and a variety of cAMP analogues. The inhibitory effect of the cAMP analogues was partially blocked by pretreatment with H89. In the presence of Ap, forskolin, or both, the expression of phospho-cAMP response element-binding protein (CREB) was increased. Ap alone had no effect on inducible cAMP early repressor (ICER) mRNA expression; however, Ap played a potentiating role in forskolin-induced ICER mRNA expression. An EMSA and immunoprecipitation assay showed that ICER binding to cAMP-response element (CRE) was increased in cells cotreated with Ap and forskolin for 3 and 8 h. Taken together, these results suggest that ICER is likely to be involved in the suppression of Ap-induced GSTP expression caused by the increase of cAMP in rat primary hepatocytes.


Efficacy review of antipsychotics.
Antipsychotic drugs have been available for more than 40 years. The invention of neuroleptics changed the overall treatment of schizophrenia dramatically. Nevertheless, the discussion on the importance and role of antipsychotic therapy still goes on. In the public as well as in the media, there remains a great deal of scepticism about the role of psychopharmacotherapy in the treatment of schizophrenia. The best way to handle this is an evidence-based approach. The efficacy of neuroleptic treatment has been proven in over 150 double-blind, controlled clinical trials. The second generation of antipsychotic drugs show similar antipsychotic properties as the classical agents, but with fewer side-effects such as extrapyramidal symptoms (EPS). The atypical antipsychotic agents have improved the quality of therapy and can also improve and support other aspects of treatment. When used with other treatment strategies, atypical antipsychotic agents could improve the overall outcome of this usually chronic disease. It is now necessary to implement these strategies as effectively as possible.


Does Depressive Affect Mediate the Relationship between Self-Care Capacity and Nutritional Status Among Rural Older Adults? : A Structural Equation Modeling Approach.
This study examined the relationships of self-care capacity and depressive affect on nutritional status and whether depressive affect mediated the relationship of self-care capacity on nutritional status. A convenience sample of 171 rural community-dwelling older adults, 65 years and above, participated. Structural equation modeling (SEM) was conducted to test a mediation model. The hypothesized SEM model was supported with adequate fit (χ2 (1) = 1.87, p = 0.17; CFI = 0.94; RMSEA = 0.07; SRMR = 0.03). SEM analysis revealed a significant positive direct effect of self-care capacity on nutritional status (γ = 0.14, p = 0.042). Significant negative direct effects were observed for self-care capacity on depressive affect (γ = -0.15, p = 0.027) and for depressive affect on nutritional status (β = -0.27, p < 0.01). Depressive affect was also observed to partially mediate the relationship of self-care capacity on nutrition status (γ = 0.04, p = 0.046). Findings highlight the importance of emotional well-being on rural older adults' nutritional status, particularly those with decreased ability to engage in self-care practices.


Structural insights into a novel Ca2+-independent PL-6 alginate lyase from Vibrio OU02 identify the possible subsites responsible for product distribution.
Alginate lyases have a wide range of industrial applications, such as oligosaccharide preparation, medical treatment, and bioconversion. Therefore, the discovery and characterization of novel alginate lyases are extremely important. PL-6 alginate lyases are classified into two groups: those with a single domain or two domains. However, only one structure of a two-domain alginate lyase has been determined to date. In this study, we characterized a novel single-domain PL-6 alginate lyase (named AlyF). According to the biochemical analysis, AlyF possesses unique features compared with other PL-6 enzymes, including (1) a Ca2+-independent catalytic mechanism and (2) a PolyG-specific cleavage specificity that predominantly produces trisaccharides. The structures of AlyF and its complexes described here reveal the structural basis for these unique features and substrate binding mechanisms, which were further confirmed using mutagenesis. More importantly, we determined the possible subsites specifying the predominantly trisaccharide products of AlyF, which may facilitate the rational design of AlyF for potential applications in preparing a single alginate oligomer.


Maintenance of lung myeloperoxidase activity in proestrus females after trauma-hemorrhage: upregulation of heme oxygenase-1.
Previous studies showed that females in the proestrus stage of the reproductive cycle maintain organ functions after trauma-hemorrhage. However, it remains unknown whether the female reproductive cycle is an important variable in the regulation of lung injury after trauma-hemorrhage and, if so, whether the effect is mediated via upregulation of heme oxygenase (HO)-1. To examine this, female Sprague-Dawley rats during diestrus, proestrus, estrus, and metestrus phases of the reproductive cycle or 14 days after ovariectomy underwent soft tissue trauma and then hemorrhage (mean blood pressure 40 mmHg for 90 min followed by fluid resuscitation). At 2 h after trauma-hemorrhage or sham operation, lung myeloperoxidase (MPO) activity and intercellular adhesion molecule (ICAM)-1, cytokine-induced neutrophil chemoattractant (CINC)-1, CINC-3, and HO-1 protein levels were measured. Plasma 17beta-estradiol concentration was also determined. The results indicated that trauma-hemorrhage increased lung MPO activity and ICAM-1, CINC-1, and CINC-3 levels in ovariectomized females. These parameters were found to be similar to sham-operated animals in proestrus female rats subjected to trauma-hemorrhage. Lung HO-1 protein level in proestrus females was increased significantly compared with female rats subjected to trauma-hemorrhage during diestrus, estrus, and metestrus phases of the reproductive cycle and ovariectomized rats. Furthermore, plasma 17beta-estradiol level was highest in proestrus females. Administration of the HO inhibitor chromium mesoporphyrin prevented the attenuation of shock-induced lung damage in proestrus females. Thus these findings suggest that the female reproductive cycle is an important variable in the regulation of lung injury following trauma-hemorrhage and that the protective effect in proestrus females is likely mediated via upregulation of HO-1.


Effect of surgical castration on risk factors for arteriosclerosis of patients with prostate cancer.
To analyze the effect of castration on risk factors for arteriosclerosis of patients with prostate cancer. Thirty patients with primary regional prostate adenocarcinoma limited to the prostate theca were selected in this study. Serum levels of testosterone (T), free testosterone (FT), dehydroepiandrosterone (DHEA), sex hormone-binding globulin (SHBG), prostatic specific antigen (PSA), triglyceride (TG), total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), apoprotein alpha(1) (APOalpha(1)) and apoprotein beta (APObeta), insulin, plasma fibrinopeptide A (FPA), plasminogen activator inhibitor-1 (PAI-1) and fibrinogen were determined just prior to, 1 week and 1, 4 and 8 months after castration. T, FT and PSA decreased significantly 1 week after castration (21.12 +/- 15.11 ng/ml vs 383.9 +/- 62.6 ng/ml, P < 0.001; 4.08 +/- 3.29 pmol/L vs 34.11 +/- 11.59 pmol/L, P < 0.001; 14.34 +/- 7.77 ng/ml vs 23.51 +/- 6.57 ng/ml, P = 0.001, respectively) and continued to decrease until reaching their lowest levels 8 months after castration. DHEA and SHBG did not undergo any changes. TG, fasting insulin and glucose, 2-hour insulin and glucose levels were significantly elevated 1 month after castration (1.84 +/- 0.61 mmol/L vs 1.30 +/- 0.40 mmol/L, P < 0.05; 18.16 +/- 5.57 mU/L vs 9.47 +/- 3.81 mU/L, P < 0.05; 4.77 +/- 0.66 mmol/L vs 3.92 +/- 0.34 mmol/L, P < 0.05; 65.52 +/- 14.78 mU/L vs 36.94 +/- 17.12 mU/L, P < 0.01; 6.98 +/- 0.79 mmol/L vs 6.01 +/- 0.23 mmol/L, P = 0.001, respectively). TC, LDL-C, FPA and PAI-1 levels were elevated 4 months after castration (6.56 +/- 0.99 mmol/L vs 5.29 +/- 0.75 mmol/L, P < 0.01; 4.09 +/- 0.86 mmol/L vs 3.04 +/- 0.15 mmol/L, P < 0.01; 3.39 +/- 1.67 nmol/L vs 1.48 +/- 0.50 nmol/L, P < 0.01; 27.02 +/- 5.98 ng/ml vs 21.78 +/- 3.16 ng/ml, P < 0.05, respectively), continuing to increase after that point. Insulin sensitive index (ISI) decreased significantly 1 month after surgery (-4.42 +/- 0.36 vs -3.50 +/- 0.39, P < 0.001), and continued to decrease from that point forward. HDL-C, APOalpha(1), APObeta and fibrinogen remained at pre-operative levels. There was a negative linear correlation between FT and TG, TC, LDL-C, PAI-1, FPA, fasting insulin and glucose, 2-hour insulin and glucose (r = -0.311, -0.384, -0.385, -0.339, -0.353, -0.381, -0.303, -0.460 and -0.395, respectively; P < 0.05). A similar phenomenon occurred with T (r = -0.308, -0.309, -0.356, -0.320, -0.430, -0.453, -0.435, -0.483 and -0.512, respectively; P < 0.05). T and FT were positively associated with ISI (r = 0.555 and 0.501; P < 0.001). At 8 months follow-up of the study subjects, we found that lower androgen levels have adverse effects on lipid metabolism, coagulative function and insulin sensitivity, related to arteriosclerosis in men.


[Diabetes mellitus in the Siberian region].
The problem of diabetes mellitus in Siberia is of utmost medico-social importance. The patterns of diabetes and abnormal carbohydrate metabolism prevalence in the Siberian and, in particular, Northern regions are related to the significant population migration to these areas. A higher incidence of carbohydrate metabolism disorders and risk factors among the migrant compared to the native population is revealed. Evaluation of diabetes prevalence, morbidity, risk factors, and of the hormonal spectrum in the Siberian residents would allow for planning and implementing further preventive actions in the framework of the multifactor prophylaxis of chronic non-infectious diseases.


How can measurement, monitoring, modeling and control advance cell culture in industrial biotechnology?
This report highlights the potential of measurement, monitoring, modeling and control (M(3) C) methodologies in animal and human cell culture technology. In particular, state-of-the-art of M(3) C technologies and their industrial relevance of existing technology are addressed. It is a summary of an expert panel discussion between biotechnologists and biochemical engineers with both academic and industrial backgrounds. The latest ascents in M(3) C are discussed from a cell culture perspective for industrial process development and production needs. The report concludes with a set of recommendations for targeting M(3) C research toward the industrial interests. These include issues of importance for biotherapeutics production, miniaturization of measurement techniques and modeling methods.


[Risk factors for mental health problems and complicated grief in bereaved families of motor vehicle accident victims].
This study examined the risk factors for mental health problems and complicated grief in bereaved families using a nationwide sample of 453 Japanese adults who had lost a family member to a motor vehicle accident within three years. The results indicate that 31.0% of participants had K6 scores > 13 and 61.0% had ICG (Inventory of Complicated Grief) scores > 26. A higher K6 score was associated with secondary victimization and support seeking, whereas a higher ICG score was associated with the death of a child. Dispute over the liability for the accident and the resulting anxiety, measured by the Japanese version of ECR (Experiences in Close Relationships), were common predictors of higher K6 and ICG scores. The results suggest that complicated grief is more dependent on the circumstances of the death, whereas mental health problems are more affected by a participant's coping after the death, implying that effective support and interventions are necessary for mental health problems and complicated grief after a violent death.


Expression of the green fluorescent protein in the oligodendrocyte lineage: a transgenic mouse for developmental and physiological studies.
We generated a transgenic mouse expressing the enhanced green fluorescent protein (EGFP) under the control of the 2'-3'-cyclic nucleotide 3'-phosphodiesterase (CNP) promoter. EGFP(+) cells were visualized in live tissue throughout embryonic and postnatal development. Immunohistochemical analysis in brain tissue and in sciatic nerve demonstrated that EGFP expression was restricted to cells of the oligodendrocyte and Schwann cell lineages. EGFP was also strongly expressed in "adult" oligodendrocyte progenitors (OPs) and in gray matter oligodendrocytes. Fluorescence-activated cell sorting allowed high-yield purification of EGFP(+) oligodendrocyte-lineage cells from transgenic brains. Electrophysiological patch clamp recordings of EGFP(+) cells in situ demonstrated that OP cells displayed large outward tetraethylammonium (TEA)-sensitive K(+) currents and very small inward currents, whereas mature oligodendrocytes were characterized by expression of large inward currents and small outward K(+) currents. The proliferation rate of EGFP(+) cells in developing white matter decreased with the age of the animals and was strongly inhibited by TEA. Oligodendrocyte development and physiology can be studied in live tissue of CNP-EGFP transgenic mice, which represent a source of pure EGFP(+) oligodendrocyte-lineage cells throughout development.


[Classification and pathogenesis of cutaneous paraneoplastic syndromes].
The associations between skin conditions and malignant tumours are reviewed and classified in two groups: indirect associations (predisposing genetic factor or carcinogenic agent), and direct associations with parallel evolutions corresponding to the true paraneoplastic syndromes. Occasional associations are also mentioned. The cutaneous paraneoplastic syndromes can be classified according to their pathogenic mechanisms, although these are mostly hypothetical, allowing those of secretory, immunological, deficient and neurovascular origins to be distinguished.


Purification and properties of human melanoma cell tissue factor.
Tissue factor (TF) is a transmembrane glycoprotein that acts as a receptor for nonactivated and activated factor VII (FVII) and triggers the coagulation cascade. TF plays an important role in hemostasis, but may also have noncoagulation functions in vascular development, angiogenesis, and tumor cell metastasis. In tumor cells, analysis of the role of TF has been hampered by the lack of purified TF. In this study, TF antigen was identified on human A375 malignant melanoma cells using flow cytometry. We further purified TF apoprotein 2,000-fold to homogeneity from A375 melanoma cells using immunoaffinity chromatography. On SDS-polyacrylamide gel electrophoresis under reduction, purified TF apoprotein gave two major protein bands corresponding to molecular weights of 53 and 34 to 36 KD. The identity of these forms of TF was confirmed by Western blotting using a polyclonal antibody against human brain TF. Under reduction, the TF antibody bound with a monomeric form of TF (53 KD), and without reduction, to several forms of TF (34 to 128 KD). Preliminary carbohydrate analysis suggested that TF is a glycoprotein and contains about 22% total carbohydrates. The coagulant activity of the purified apoprotein was reconstituted by the addition of phospholipids. The effects of varying concentrations (0 to 8 microg) of polyclonal antibodies to TF and FVII on TF procoagulant activity were studied. Both antibodies inhibited more than 70% of the procoagulant activity of TF in an FX activation assay. The complex formation between purified TF apoprotein and FVIIa was demonstrated by using an enzyme-linked immunosorbent assay. TF formed a complex with FVIIa in a concentration-dependent and saturable manner. We conclude that in human melanoma cells, TF occurs in monomeric and heterodimeric forms and appears to have similar properties as reported for TF from other sources.


Long-Term Efficacy, Visual Performance and Patient Reported Outcomes with a Trifocal Intraocular Lens: A Six-Year Follow-up.
(1) Background: To evaluate the efficacy at 6 years postoperative after the implantation of a trifocal intraocular lens (IOL) AT Lisa Tri 839MP. The secondary objective was to evaluate the contrast sensitivity defocus curve (CSDC), light distortion analysis (LDA), and patient reported outcomes (PROs). (2) Methods: Sixty-two subjects participated in phone call interviews to collect data regarding a visual function questionnaire (VF-14), a patient reported spectacle independence questionnaire (PRSIQ), and questions related to satisfaction and decision to be implanted with the same IOL. Thirty-seven of these subjects were consecutively invited to a study visit for measurement of their visual acuity (VA), CSDC, and LDA. (3) Results: The mean monocular distance corrected VA was -0.05, 0.08, and 0.05 logMAR at far and distances of 67 cm and 40 cm, respectively. These VAs were significantly superior to those reported in previous literature (p < 0.05). The total area under the CSDC was 2.29 logCS/m-1 and the light distortion index 18.82%. The mean VF-14 score was 94.73, with 19.4% of subjects requiring spectacles occasionally for near distances, and 88.9% considering the decision of being operated again; (4) Conclusions: Long-term AT LISA Tri 839MP IOL efficacy results were equal or better than those reported 12 months postoperatively in previous studies. The spectacle independence and satisfaction rates were comparable to those reported in short-term studies.


Co-infection with Campylobacter and rotavirus in less than 5 year old children with acute gastroenteritis in Nepal during 2017-2018.
Diarrhoea, although easily curable, is a global cause of death for a half million children every year. Rotavirus and Campylobacter are the most common etiological agents of diarrhoea in children less than 5 years of age. However, in Nepal, these causative agents are not routinely examined for the diagnosis and treatment. The main objective of this study was to determine Campylobacter co-infection associated with rotavirus diarrhoea in children less than 5 years of age. A cross-sectional study was conducted at Kanti Children's Hospital (KCH), Kathmandu, Nepal from November 2017 to April 2018. A total of 303 stool specimens from children affected with diarrhoea were processed to detect rotavirus using a rapid rotavirus antigen detection test kit, and Campylobacter by microscopy, culture and biochemical tests. Antibiotic susceptibility tests of Campylobacter isolates were performed according to European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines 2015. Of 303 samples, 91 (30.0%) were positive for co-infection with rotavirus and Campylobacter. Rotavirus mono-infection was detected in 61 (20.1%), and Campylobacter mono-infection was detected in 81 (26.7%) samples. Patient's age, month of infection, untreated water and frequent soil contact were the major risk factors for infections. Clinical features such as > 9 loose motions per day, fever, vomiting, mild to moderate dehydration, diarrhea persisting 6-9 days and presence of mucus in stool were significant (p < 0.05) clinical features, and were more severe in coinfection compared to mono-infections in multivariate analysis. The study shows a high rate of rotavirus and Campylobacter coinfection in children with diarrhoea. Diagnosis based management of diarrhoeal cases can guide the specific treatment.


Adolescent psychiatric inpatients' self-reported reasons for cutting themselves.
To ascertain (1) whether male and female adolescent (13-17 years old) psychiatric inpatients endorse comparable reasons for cutting themselves and (2) whether these reasons are correlated with selected psychosocial characteristics of the adolescents, self-reported depression, and hopelessness, the Self-Injury Motivation Scale II (SIMS-II), the Beck Depression Inventory-II, and the Beck Hopelessness Scale were administered to 19 (38%) male and 31 (62%) female adolescents who had cut themselves. Independent t tests found that none of the SIMS-II subscale scores was differentiated by sex, but the Beck Depression Inventory-II total score was significantly correlated with the SIMS-II total, Affect Modulation, Desolation, and Punitive Duality subscale scores. The results are discussed as indicating that male and female adolescent inpatients endorse comparable reasons for cutting themselves and that self-reported depression is positively associated with the number and intensity of different motivations for cutting oneself.


Chicken chemokine receptors in T cells isolated from lymphoid organs and in splenocytes cultured with concanavalin A.
Chemokine receptors guide immune cells to specific organs during health and disease. The mRNA content of the chemokine receptors CCR2, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CXCR4, CXCR5, and CX3CR1 in CD4(+) cells (T-helper cells) isolated from blood, bursa, cecal tonsil, spleen, and thymus and in CD8(+) cells (T-cytotoxic cells) isolated from blood, cecal tonsil, spleen, and thymus were investigated. The CD4(+) cells isolated from thymus had the highest amount of CCR7 and CCR8 mRNA. The CD4(+) cells isolated from bursa, cecal tonsil, and thymus had the highest amount of CCR5 mRNA. The CD4(+) cells isolated from cecal tonsils had the highest amount of CCR9 mRNA. The CD4(+) cells isolated from bursa and thymus had the highest amount of CXCR5 mRNA. The CD8(+) cells isolated from cecal tonsil had the highest mRNA amount of all receptors studied except CCR9 and CX3CR1. The CD4(+) cells treated with concanavalin A had increased CCR2, CCR4, CCR7, CCR8, and CXCR5 mRNA amounts at 24 h of stimulation. The CD8(+) cells treated with concanavalin A had increased CCR4 mRNA at 72 h, increased CCR6 mRNA at 24 h, and decreased CCR8 and CXCR4 mRNA at 24 h of stimulation.


Geminivirus C2 protein might be the key player for geminiviral co- option of SCF-mediated ubiquitination.
Viruses are obligate intracellular parasites, and need to create a suitable cell environment for viral propagation to complete their life cycle. In order to achieve this, viruses must usurp or interfere with the cellular machinery. Ubiquitination, a post-translational modification that controls numerous cellular processes, has proven to be a common target for viruses. Recently, geminivirus C2 protein has been shown to interact with the CSN complex and disrupt its activity over CULLIN1, interfering with the function of the CULLIN1-based SCF ubiquitin E3 ligases. Interestingly, overexpression of a given F-box protein may circumvent the general SCF malfunction caused by C2. This result raises the tantalizing idea that geminiviruses might be not only hampering, but also redirecting the activity of SCF complexes, thus co-opting the SCF-mediated ubiquitination pathway. We hypothesize that the mechanism of C2-facilitated co-option of SCF-mediated ubiquitination might not be exclusive for geminiviruses, but rather a common strategy for viruses.


Prediction of graft viability from non-heart-beating donor pigs using hepatic microdialysate hypoxanthine levels.
Non-heart-beating donors (NHBDs) are not yet acceptable in orthotopic liver transplantation (OLTX) because of the high frequency of primary graft nonfunction. In this study, we aimed to develop a new predictive method of graft viability in OLTX from NHBDs. (1) Pigs were subjected to 15 min of hepatic ischemia and reperfusion (I/R). (2) Porcine OLTX was performed using grafts obtained from NHBDs subjected to in situ warm ischemia (0, 30, and 60 min). During both operations, hepatic hypoxanthine levels were measured by a microdialysis method. In the I/R model, hypoxanthine accumulated during ischemia and decreased after reperfusion, whereas marked xanthine and uric acid production were observed after reperfusion. In the NHBDs model, all of the 0-min group, 6 of 13 pigs in the 30-min group, and 1 of 6 pigs in the 60-min group survived more than 7 days. Significant increases of hypoxanthine levels were seen dependent on warm ischemic time. In the 30-min group, hypoxanthine levels were significantly higher in the pigs that died than in those that survived, and correlated with aspartate aminotransferase, lactate dehydrogenase, and adenosine triphosphate levels of recipients. Histological examination revealed that graft injury was severe in the pigs with higher hypoxanthine levels. It is suggested that hepatic microdialysate hypoxanthine levels during warm ischemia in NHBDs were correlated with graft viability in the recipient. By using of this technique, prediction of the graft viability may be possible during donor operation.


[Contribution of PCR for detection and identification of intestinal microsporidia in HIV-infected patients].
Intestinal microsporidiosis are among the most frequent opportunistic diseases in immunocompromised subjects. This study aimed to evaluate the contribution of PCR for a better detection and species identification of microsporidia in stool specimens of HIV-infected patients. Stool samples obtained from 119 HIV-infected Tunisian subjects were screened for intestinal microsporidiosis by light microscopy using Weber's modified Trichrome stain and by a PCR method using universal primers V1/PMP2 which amplified a common fragment of the small subunit rRNA gene of microsporidia. The obtained PCR products were then sequenced using an ABI PRISM 377 DNA sequencer. The results showed a better sensitivity of PCR in the detection of microsporidia with an infection rate of 14.3% significantly higher than that of 6.7% obtained by light microscopy (p=0.03). As previously described, intestinal microsporidiosis was associated with low CD4 cell counts; 23.9% infection rate in patients having CD4 cell count under 200/mm(3) against 5.6% in patients with higher CD4 cell count (p=0.008). The sequencing of 15 out of the 17 positive PCR products has confirmed in all cases the species identified based on the PCR fragment size i.e., 250pb for Enterocytozoon bieneusi (seven cases) and about 270pb for Encephalitozoon intestinalis (nine cases); one case revealed a double infection. PCR proved to be more effective than classical Trichrome stain for the diagnosis of intestinal microsporidiosis. Moreover, the ability of PCR to identify the species involved could also be useful for cases management.


Physiological studies of biosynthetic indole excretion in Bacillus alvei.
Bacillus alvei excretes indole during early exponential growth in acid-hydrolyzed casein medium. l-Threonine is the amino acid responsible for "early" indole excretion, and the amount of indole excreted is directly related to the amount of l-threonine in the medium. "Early-indole" excretion can be prevented by the continuous addition of serine (3.1 mumoles per ml per hr) or by substituting a mutant with an impaired ability to degrade serine. The addition of serine to a culture during the period of indole excretion halts the excretion and stimulates indole utilization. Threonine is a competitive inhibitor of serine (K(i) = 0.6 m) in the tryptophan synthetase B reaction. The internal tryptophan concentration increases during the period of indole excretion, suggesting that threonine acts by increasing the activity of the tryptophan pathway. This view is supported by experiments demonstrating that anthranilic acid and indoleacrylic acid also stimulate indole excretion. A metabolic explanation is offered and discussed.


Two-color fluorescent (near-infrared and visible) triphasic perfluorocarbon nanoemuslions.
Design and development of a new formulation as a unique assembly of distinct fluorescent reporters with nonoverlapping fluorescence spectra and a F19 magnetic resonance imaging agent into colloidally and optically stable triphasic nanoemulsion are reported. Specifically, a cyanine dye-perfluorocarbon (PFC) conjugate was introduced into the PFC phase of the nanoemulsion and a near-infrared dye was introduced into the hydrocarbon (HC) layer. To the best of our knowledge, this is the first report of a triphasic nanoemulsion system where each oil phase, HC, and PFC are fluorescently labeled and formulated into an optically and colloidally stable nanosystem. Having, each oil phase separately labeled by a fluorescent dye allows for improved correlation between in vivo imaging and histological data. Further, dual fluorescent labeling can improve intracellular tracking of the nanodroplets and help assess the fate of the nanoemulsion in biologically relevant media. The nanoemulsions were produced by high shear processing (microfluidization) and stabilized with biocompatible nonionic surfactants resulting in mono-modal size distribution with average droplet size less than 200 nm. Nanoemulsions demonstrate excellent colloidal stability and only moderate changes in the fluorescence signal for both dyes. Confocal fluorescence microscopy of macrophages exposed to nanoemulsions shows the presence of both fluorescence agents in the cytoplasm.


Diagnosis and treatment of recurrent high-grade astrocytoma.
High-grade gliomas represent a significant source of cancer-related death, and usually recur despite treatment. In this analysis of current brain tumor medicine, we review diagnosis, standard treatment, and emerging therapies for recurrent astrocytomas. Difficulties in interpreting radiographic evidence, especially with regard to differentiating between tumor and necrosis, present a formidable challenge. The most accurate diagnoses come from tissue confirmation of recurrent tumor, but a combination of imaging techniques, such as magnetic resonance spectroscopy imaging, may also be relevant for diagnosis. Repeat resection can prolong life, but repeat irradiation of the brain poses serious risks and results in necrosis of healthy brain tissue; therefore, reirradiation is usually not offered to patients with recurrent tumors. We describe the use of conventional radiotherapy, intensity-modulated radiotherapy, brachytherapy, radiosurgery, and photodynamic therapy for recurrent high-grade glioma. The use of chemotherapy is limited by drug distribution and toxicity, but the development of new drug-delivery techniques such as convection-enhanced delivery, which delivers therapeutic molecules at an effective concentration directly to the brain, may provide a way to reduce systemic exposure to cytotoxic agents. We also discuss targeted therapies designed to inhibit aberrant cell-signaling pathways, as well as new experimental therapies such as immunotherapy. The treatment of this devastating disease has so far been met with limited success, but emerging knowledge of neuroscience and the development of novel therapeutic agents will likely give patients new options and require the neuro-oncology community to redefine clinical trial design and strategy continually.


Fentanyl decreases catecholamine metabolism measured by in vivo voltammetry in the rat locus coeruleus.
The objective of this study was to investigate under controlled conditions the effects of fentanyl on the rat locus coeruleus catechol oxidation current. Using differential normal pulse voltammetry combined with electrochemically treated carbon fiber electrodes to measure the catechol oxidation current, catecholamine metabolism can be reliably monitored. Male Sprague-Dawley rats weighing 500-600 g had carbon fiber electrodes implanted into the locus coeruleus under halothane - O2 - air anesthesia with controlled ventilation and muscle relaxation. Experiments consisted of four groups of rats given the following treatments: (A) saline (n = 6); (B) fentanyl, 10 micrograms.kg-1 i.v. (n = 6); (C) naloxone, 800 micrograms.kg-1 i.v. followed 2 min later by fentanyl, 10 micrograms.kg-1 (n = 5); (D) clonidine, 200 micrograms.kg-1 i.p. (n = 6). There was no significant change in the catechol oxidation current following saline. Fentanyl produced a significant (ANOVA, p less than 0.05) decrease in the catechol oxidation current (maximum 32 min postinjection was 75.8 +/- 4.6% of baseline). This decrease was prevented by a prior injection of naloxone. Clonidine produced a significant decrease in catechol oxidation current (maximum 40 min postinjection was 54.1 +/- 7.0% of baseline). Systolic blood pressure was significantly decreased following clonidine and there were no significant changes in arterial blood gases throughout the experiments. The alpha 2-adrenergic agonist clonidine and the opioid fentanyl produced a decrease in locus coeruleus catechol oxidation current measured by in vivo voltammetry, which monitors catecholamine turnover.


[Anti-aging medicine, a science-based, essential medicine].
Anti-aging medicine is booming. It enters more and more in the programs of universities. Its hormone and nutritional tests and therapies rely on numerous scientific studies, including double-blind placebo-controlled randomized studies. Its methods are often innovative to obtain more information or benefits with greater safety. The fundamental purpose of anti-aging medicine is to optimize health and the quality of life, and through this, make the physical appearance more youthful. Well-chosen and well-dosed these treatments should not increase the risk of age-related diseases such as cancer and cardiovascular diseases, but on the contrary decrease it by the preventive aspect of the treatments. Opponents to anti-aging medicine fail in collecting valid scientific arguments. Their insistence on maintaining a society of elderly people looking and feeling as elderly people rather than actively participating in searching for ways to attenuate aging is harmful to all who follow them, to themselves in the first place.


Prolongation of the P-300 latency associated with hydrogen sulfide exposure.
Three patients exposed to hydrogen sulfide developed persistent cognitive impairment, as suggested by the P-300 event-related potential and measured by neuropsychological testing. Routine neurological and physical examinations were unremarkable, although the patients were sufficiently impaired so as to be unable to work. The P-300 event-related potential and neuropsychological testing proved to be important in the detection of cognitive dysfunction following acute hydrogen sulfide exposure. The three patients with neurocognitive dysfunction were acutely exposed to hydrogen sulfide. The incidents occurred independently and under different circumstances. Each patient was evaluated at the Northern California Occupational Health Clinic, San Francisco, examined neurologically and neuropsychologically, and evaluated with a P-300 event-related potential. Each patient had persistent neurological symptoms, neuropsychological deficit, and abnormally prolonged P-300 latencies.


Neonatal Sepsis and Associated Factors Among Newborns in Woldia and Dessie Comprehensive Specialized Hospitals, North-East Ethiopia, 2021.
Neonatal sepsis is a clinical illness characterized by infection-related signs and symptoms in the first month of life, with or without bacteremia. Septicemia, meningitis, pneumonia, arthritis, osteomyelitis, and urinary tract infections are all examples of systemic illnesses that can affect newborns. Hence, the main aim of this study was to assess the prevalence and factors associated with neonatal sepsis among newborns in Woldia and Dessie Comprehensive Specialized Hospitals, northeast Ethiopia, from January 1 to July 30, 2021. This institution-based cross-sectional study was conducted from January 1 to July 30, 2021, on 344 randomly selected neonates who visited the hospital. A systematic random sampling technique was used to select samples, and data were collected using a pre-tested standardized questionnaire. For data entry and analysis, Epi Data version 4.1 and SPSS version 24 applications were used, respectively. The goodness-of-fit was tested by the Hosmer-Lemeshow statistic test. Bivariable and multivariable binary logistic regressions were used to identify associated factors at a 95% confidence interval. Significance was considered at p-value <0.05. In this study, the prevalence of neonatal sepsis was 79.4% (95% CI: 75.2-83.6%). Maternal UTI/STI history [AOR: 3.1; 95% CI (1.5-7.1)], gestational age <37 weeks [AOR: 4.4; 95% CI (1.0-8.9)], PROM [AOR: 4.9; 95% CI (2.5-6.8)], and new-born resuscitation history [AOR: 2.3; 95% CI (1.5-4.3)] were all significantly associated with neonatal sepsis. This study indicates that the proportion of neonatal sepsis is high. A history of maternal PROM, being a preterm neonate, a history of maternal UTI/STIs, and having received resuscitation at birth were identified as risk factors for neonatal sepsis.


Assessing the public's views in research ethics controversies: deliberative democracy and bioethics as natural allies.
In a liberal democracy, policy decisions regarding ethical controversies, including those in research ethics, should incorporate the opinions of its citizens. Eliciting informed and well-considered ethical opinions can be challenging. The issues may not be widely familiar and they may involve complex scientific, legal, historical, and ethical dimensions. Traditional surveys risk eliciting superficial and uninformed opinions that may be of dubious quality for policy formation. We argue that the theory and practice of deliberative democracy (DD) is especially useful in overcoming such inadequacies. We explain DD theory and practice, discuss the rationale for using DD methods in research ethics, and illustrate in depth the use of a DD method for a longstanding research ethics controversy involving research based on surrogate consent. The potential pitfalls of DD and the means of minimizing them as well as future research directions are also discussed.


The spindle position checkpoint requires positional feedback from cytoplasmic microtubules.
The objective of mitosis is to provide a copy of the genome to each progeny of a cell division. This requires the separation of duplicate chromatids by the spindle apparatus and the delivery of one set of chromosomes to each of the daughter cells. In budding yeast, the fidelity of chromosome delivery depends on the spindle position checkpoint, which prolongs mitosis until one end of the anaphase spindle arrives in the bud. Here we tested the hypothesis that the activity of the spindle position checkpoint depends on persistent interactions between cytoplasmic microtubules and the mother-bud neck, the future site of cytokinesis. We used laser ablation to disrupt microtubule interactions with the bud neck, and we found that loss of microtubules from the neck leads to mitotic exit in a majority of checkpoint-activated cells. Our findings suggest that cytoplasmic microtubules are used to monitor the location of the spindle in the dividing cell and, in the event of positioning errors, relay a signal to inhibit mitotic exit until the spindle is appropriately positioned.


A far ultraviolet polarization analyzer for rocket use.
A rocket version of a far uv polarization analyzer has been constructed to measure the polarization of resonantly scattered sunlight. The rotating, eight-plate, LiF polarization analyzer has principal transmittances which vary from k(1) = 0.088 and k(2) = 0.0037 at H Lyman-alpha to k(1) = 0.623 and k(2) = 0.075 at 2500 A. The region from 1200 A to 2000 A is divided into four intervals by using filters of CaF(2), BaF(2), and Al(2)O(3) in conjunction with the solar blindness of the CsI photocathode. The calculated polarizations, which are a direct consequence of the Zeeman effect, are given for some of the uv multiplets which could contribute to the uv day airglow in planetary atmospheres. Departures from the calculated polarizations may be due to the effects of multiple scattering, collisional excitation, and hyperfine structure. The observed polarization of the sum of the lines which constitute a multiplet is dependent upon the relative population (a measure of the temperature) of the ground states of the multiplet.


Targeting the Adenosinergic Axis in Chronic Lymphocytic Leukemia: A Way to Disrupt the Tumor Niche?
Targeting adenosine triphosphate (ATP) metabolism and adenosinergic signaling in cancer is gaining momentum, as increasing evidence is showing their relevance in tumor immunology and biology. Chronic lymphocytic leukemia (CLL) results from the expansion of a population of mature B cells that progressively occupies the bone marrow (BM), the blood, and peripheral lymphoid organs. Notwithstanding significant progress in the treatment of these patients, the cure remains an unmet clinical need, suggesting that novel drugs or drug combinations are needed. A unique feature of CLL is its reliance on micro-environmental signals for proliferation and cell survival. We and others have shown that the lymphoid niche, an area of intense interactions between leukemic and bystander non-tumor cells, is a typically hypoxic environment. Here adenosine is generated by leukemic cells, as well as by cells of myeloid origin, acting through autocrine and paracrine mechanisms, ultimately affecting tumor growth, limiting drug responses, and skewing the immune cells towards a tolerant phenotype. Hence, understanding the mechanisms through which this complex network of enzymes, receptors, and metabolites functions in CLL, will pave the way to the use of pharmacological agents targeting the system, which, in combination with drugs targeting leukemic cells, may get us one step closer to curing these patients.


Importance of recognizing and managing delirium in intensive care unit.
Delirium is an acute and fluctuating change in mental status, with inattention and altered levels of consciousness. It is a common comorbidity in intensive care units (ICU), resulting in delayed withdrawal of mechanical ventilation, prolonged length of stay in ICU, increased ICU mortality and impaired long-term cognitive function of the survivors. Narcotic or psychoactive medication is one of the major risk factors that contribute to ICU delirium. Surveys conducted in several countries indicated that delirium in ICU was inadequately monitored, underdiagnosed and lacked standardized treatment. In order to improve the prevention and treatment of ICU delirium, it is imperative that the ICU professionals should enrich their knowledge about this comorbidity, familiarize themselves with its screening and management, as well as standardize the administration of narcotic and psychoactive medications.


Efficient fabrication of ordered alumina through-hole membranes using a TiO2 protective layer prepared by atomic layer deposition.
Ordered alumina through-hole membranes were obtained by a combination of the anodization of Al, formation of a TiO2 protective layer, and subsequent etching. Two-layered anodic porous alumina materials composed of TiO2-coated and noncoated alumina were prepared by the combination of the anodization of Al and the formation of a TiO2 protective layer by atomic layer deposition (ALD). The obtained two layers of anodic porous alumina have different solubilities because the TiO2 thin layer formed by ALD acts as a protective layer that prevents the dissolution of the alumina layer during wet etching of the sample in an etchant. After the selective dissolution of the bottom layer of porous alumina without the TiO2 layer, an ordered alumina through-hole membrane could be detached from the Al substrate. This process allows the repeated preparation of ordered alumina through-hole membranes from a single Al substrate. By this process, ordered alumina through-hole membranes with large interhole distances could also be obtained. The obtained alumina through-hole membrane can be used in various applications.


Physician-Perceived Predictive Factors for the Effectiveness of Drugs for Treating Cancer Dyspnea: Results of a Nationwide Survey of Japanese Palliative Care Physicians.
Background: Dyspnea is a common and distressing symptom in patients with advanced cancer. Opioids, benzodiazepines, and corticosteroids are commonly prescribed pharmacological treatments for cancer dyspnea. Objective: The objective of this survey was to investigate physician-perceived predictive factors for the effectiveness of opioids, benzodiazepines, and corticosteroids in treating cancer dyspnea. Design: This study involves a nationwide survey using self-report questionnaires. Setting/Subjects: Random sampling selected 268 Japanese certified palliative care physicians in Japan. Measurements: We inquired about the 12 physician-perceived predictive factors for the effectiveness of drugs (opioids, benzodiazepines, and corticosteroids) in treating cancer dyspnea. Results: The frequently selected physician-perceived predictive factors for the effectiveness of opioids were tachypnea, respiratory effort, opioid naive, Eastern Cooperative Oncology Group Performance Status 0-2, multiple lung tumors, dry cough, pleural effusion, and pleural lesion. Benzodiazepines were predicted to be effective against dyspnea in patients with depression and severe anxiety. Meanwhile, corticosteroids were predicted to be effective against dyspnea in patients with lymphangitis carcinomatosa, superior vena cava syndrome, major airway obstruction, and audible wheezing. Japanese palliative care physicians anticipate that different drug classes will be effective for treating dyspnea in patients with specific factors. Conclusions: Japanese palliative care physicians expect that different drugs will be effective for dyspnea in patients with specific predictive factors. Future prospective studies are required to assess the effectiveness of each drug class against specific dyspnea.


Effect of cyanocobalamin (vitamin B12) on paraquat-induced brain injury in mice.
The goal of this study was to evaluate the neuroprotective effects of vit B12 on paraquat-induced neurotoxicity. Thirty-six male mice were randomly divided into six groups. Three groups were treated intraperitoneally with paraquat (10 mg/kg) twice a week (with a 3-day interval) for 3 weeks. Normal saline, vit B12 (1 mg /kg), or vit C (50 mg/kg) was injected 30 min before paraquat administration. Other groups only received normal saline (control), vit B12, or vit C in the same protocol. Motor performance and coordination were assayed by challenging beam traversal, pole, open field, and rotarod tests. The hippocampus and serum samples were isolated to evaluate the oxidative stress (GSH and ROS), apoptosis (caspase 3), and inflammatory markers (TNF-α and IL-1β). Administration of paraquat leads to induction of motor deficits, which were improved by treatment with vit B12. In addition, vit B12 could prevent oxidative damage, apoptosis, and inflammation caused by paraquat. It seems that vit B12 could be a novel therapeutic agent in the management of paraquat induced-neurotoxicity.


Prevention of angiotensin II-induced cardiac remodeling by angiotensin-(1-7).
Cardiac remodeling, which typically results from chronic hypertension or following an acute myocardial infarction, is a major risk factor for the development of heart failure and, ultimately, death. The renin-angiotensin system (RAS) has previously been established to play an important role in the progression of cardiac remodeling, and inhibition of a hyperactive RAS provides protection from cardiac remodeling and subsequent heart failure. Our previous studies have demonstrated that overexpression of angiotensin-converting enzyme 2 (ACE2) prevents cardiac remodeling and hypertrophy during chronic infusion of angiotensin II (ANG II). This, coupled with the knowledge that ACE2 is a key enzyme in the formation of ANG-(1-7), led us to hypothesize that chronic infusion of ANG-(1-7) would prevent cardiac remodeling induced by chronic infusion of ANG II. Infusion of ANG II into adult Sprague-Dawley rats resulted in significantly increased blood pressure, myocyte hypertrophy, and midmyocardial interstitial fibrosis. Coinfusion of ANG-(1-7) resulted in significant attenuations of myocyte hypertrophy and interstitial fibrosis, without significant effects on blood pressure. In a subgroup of animals also administered [d-Ala(7)]-ANG-(1-7) (A779), an antagonist to the reported receptor for ANG-(1-7), there was a tendency to attenuate the antiremodeling effects of ANG-(1-7). Chronic infusion of ANG II, with or without coinfusion of ANG-(1-7), had no effect on ANG II type 1 or type 2 receptor binding in cardiac tissue. Together, these findings indicate an antiremodeling role for ANG-(1-7) in cardiac tissue, which is not mediated through modulation of blood pressure or altered cardiac angiotensin receptor populations and may be at least partially mediated through an ANG-(1-7) receptor.


[Psoriasis therapy with retinoid].
Retinoids are effective in all forms of psoriasis and can be used alone or in combination with other therapeutic measures (anthralin, corticoids, PUVA, SUP). The effect of retinoids on psoriasis, in the final analysis still open to question, is discussed with regard to three parameters of psoriatic epidermis (enhanced proliferation, disturbance in differentiation, leucotaxis).


Deprescribing guidelines: An international symposium on development, implementation, research and health professional education.
Deprescribing is a clinically important and feasible innovation that ensures medication efficacy, reduces harms, and mitigates polypharmacy. It involves reducing doses or stopping medications that are not useful, no longer needed, or which may be causing harm. It may also involve changing to a safer agent or using non-pharmacological approaches for care instead. Clinical guidelines combined with behaviour changes (of health care providers (HCPs), the public, and health care decision-makers) are needed to integrate deprescribing into routine practice. Using rigorous international standards, the Bruyère Research Institute Deprescribing Guidelines research team validated a ground-breaking deprescribing guideline methodology and developed or co-developed 5 evidence-based deprescribing guidelines. In March 2018, the team hosted an international symposium convening HCPs, researchers, public agencies, policymakers, and patient advocates in Ottawa, Ontario, Canada. This 3-day symposium aimed to facilitate knowledge exchange amongst guideline developers, users, and the public; initiate partnerships and collaborations for new deprescribing guideline recommendations and effectiveness research; and to continue work on HCP deprescribing education activities. An interprofessional planning committee developed an overall agenda, and small groups worked on session objectives and formats for different components: methods for rigorous deprescribing guideline development, implementation experiences, research/evaluation experiences and educational needs. Through a series of keynote speakers, panel discussions, and small working groups, the symposium provided a forum for participants to meet one another, learn about their different experiences with deprescribing guidelines, and develop collaborations for future initiatives. One hundred thirty participants, from 10 countries and representing over 100 institutions and organizations took part. Symposium proceedings are presented in this issue of RSAP for sharing with the wider community engaged in the care of patients with problematic polypharmacy.


The role of unfolded protein response in differentiation of mammary epithelial cells.
The accumulation of misfolded proteins in the ER provokes ER stress by increasing the demand for energy, chaperones, and other proteins that are needed to fold client proteins or to degrade unfoldable secretory cargo. This stress activates a signaling network called the unfolded protein response (UPR). However, recent accumulated data suggested that the UPR also provides important signals for regulating cell differentiation and maturation. However, the relationship between UPR and mammary gland development has not been fully elucidated. To define the involvement of the UPR in mammary gland development, mammary glands were collected from non-pregnant mice, at days 5, 10 and 15 of pregnancy, at days 1 and 7 of lactation, and the expression patterns of UPR-related genes were determined by real-time PCR. We found that the mRNA expression of ATF4 and XBP1 significant increased during pregnancy. Moreover, we found that both ATF4 and XBP1 proteins are expressed in mammary epithelial cells by immunohistological analysis. In order to know the role of ATF4 and XBP1 in the differentiation of mammary epithelial cell, we performed gene knockdown experiment using HC11 cells. We found that ATF4 or XBP1 knockdown suppressed the mRNA expression of beta-casein and lactogenic hormone receptor in differentiating HC11 cells. Our results demonstrate that XBP1 and ATF4, which are UPR-related transcription factors, directly or indirectly participate in cell differentiation mechanisms through the regulation of the expression of lactogenic hormone receptors in mouse mammary epithelial cells.


A Noninvasive Miniaturized Transcutaneous Oxygen Monitor.
Transcutaneous monitoring is a noninvasive method to continuously measure the partial pressures of oxygen and carbon dioxide that diffuse through the skin and correlate closely with changes in blood gases. However, the contemporary commercially available electrochemical-based technology requires a heating mechanism and a bulky, corded, and expensive sensing unit. This study aims to demonstrate a prototype noninvasive, miniaturized monitor that uses luminescence-based technology to measure the partial pressure of transcutaneous oxygen, a surrogate of the partial pressure of arterial oxygen. To be able to build a robust measurement system, we conducted experiments to understand the temperature and humidity dependence of oxygen-sensitive platinum-porphyrin films. We performed a detailed analysis of both intensity and lifetime measurement techniques. To verify the performance, we tested the prototype in a small ex-vivo experiment involving three healthy human volunteers. We measured variations in the partial pressure of transcutaneous oxygen values due to pressure-induced arterial and venous occlusions on the volunteers' fingertips. The system resolves changes in the partial pressure of oxygen from 0 to 418 mmHg in the lab bench-top testing, covering the medically relevant range of 50-150 mmHg. Under fixed humidity, temperature, and the partial pressure of oxygen conditions, the sensor shows a 2% drift over 60 hours. The prototype consumes 9 mW of power from a 2.2 V external DC power supply.


Muscarinic potentiation of GABA(A) receptor currents is gated by insulin signaling in the prefrontal cortex.
Cholinergic neurotransmission and insulin signaling in cognitive areas, such as the prefrontal cortex (PFC), play a key role in regulating learning and memory. However, the cellular mechanisms by which this regulation occurs are unclear. Because GABAergic inhibition in the PFC controls the timing of neuronal activity during cognitive operations, we examined the potential regulation of GABA transmission by cholinergic and insulin signaling in PFC pyramidal neurons. Activation of muscarinic acetylcholine receptors (mAChRs) with carbachol produced an enhancement of GABA(A) receptor currents in acutely dissociated cells after a short treatment with insulin. Inhibiting phosphoinositide-3 kinase (PI3K), a downstream target of insulin signaling, eliminated this effect as well as the carbachol-induced enhancement of GABAergic miniature IPSC amplitudes in PFC slices. The muscarinic potentiation of GABA(A) currents was blocked by PKC inhibitors, broad-spectrum protein tyrosine kinase inhibitors, and specific inhibitors of the nonreceptor tyrosine kinase Src. Additionally, muscarinic receptors in PFC slices activated PKC and the focal adhesion kinase Pyk2 (a potential molecular link between PKC and Src) in a PI3K-dependent manner. Together, our results show that mAChR activation in PFC pyramidal neurons enhances GABA(A) receptor functions through a PKC-dependent, Src-mediated signaling cascade that is gated by an insulin/PI3K pathway. Given the significance of GABAergic transmission in regulating PFC functions, our results provide a novel mechanism for understanding the role of cholinergic systems and insulin signaling in learning and memory.


The DNA restriction endonuclease of Escherichia coli B. II. Further studies of the structure of DNA intermediates and products.
The DNA intermediates and final products formed by the Type I restriction endonuclease, EcoB, were further characterized. DNA cleaved on only one strand (hemi-restricted DNA) contains gaps of approximately 70-100 nucleotides, while the fully restricted products contain 3'-single-stranded tails averaging approximately 70-100 nucleotides for each strand cleaved. The gaps and tails are formed with the release of an equal number of nucleotides as small oligonucleotides that are soluble in acid. After purification, neither the hemi-restricted nor the fully restricted DNAs are cleaved again by EcoB. There is no apparent specificity for which strand of a duplex is initially cleaved by EcoB, nor is there specificity with respect to the composition of the 3'-terminal nucleotide formed on the DNA or the 3'- or 5'-terminal nucleotides of the acid-soluble oligonucleotides released during DNA cleavage. The structure formed at the 5' terminus of the DNA product which blocks phosphorylation by T4 polynucleotide kinase remains unknown, but its removal with phage lambda exonuclease allows at least some reutilization of recognition sites by EcoB as well as phosphorylation of the newly formed 5' termini. To explain the complex mechanism of this enzyme, it is suggested that the unidentified 5'-tails prevent wasteful rerestriction from occurring, whereas the 3'-single-stranded tails create DNA which, when nonhomologous to chromosomal DNA, cannot be rescued because such tails are not substrate for DNA polymerases. However, when homologous chromosomal DNA exists, the randomly cleaved large fragments with these tails can easily be assimilated by recA-mediated genetic recombination, thus stimulating DNA exchange between related organisms.


Influence of Psychological Disorders on the Functional Outcomes in the Survivors of Ischemic Stroke.
Stroke has the greatest disabling impact of any chronic disease. The survivors of stroke, experience an average of 2.38 stroke related emotional, and behavioral components. In this study we decided to find out the frequency of psychological disorders and its association with impairment of daily living. We assessed the association of psychological symptoms after stroke and study their impact on physical functional recovery. This is a hospital based observational cross-sectional study to assess the impact of early psychological symptoms on outcomes for patients with new or recurrent stroke. All subjects were evaluated in detail as per the case report for neurologic manifestations, psychological symptoms, stroke risk factors, complications and comorbidities. Relevant clinical information were recorded using the Duke Severity of Illness Scale. General Health Questionnaire 28 (GHQ28), Modified Barthel Index along with statistical tests like Chi square test was used in the study. Depression is the most prevalent psychological disorder as assessed by GHQ28 and Present State Examination in survivors of acute ischemic stroke. In survivors of acute ischemic stroke diabetes mellitus is most significantly associated with impairment of daily living. Hypertension, diabetes mellitus, obesity, smoking and dyslipidemia were found to be the most important risk factors in the present study. Hypertension was the most prevalent risk factor in these cases. But diabetes was found to be statistically significantly associated with impairment of daily living in the survivors of acute ischemic stroke. Present study entitled as Influence of psychological disorders on the functional outcomes in the survivors of acute ischemic stroke included 50 cases of survivors of acute ischemic stroke. Male to female ratio was 12:13. Mean ages in cases was 66.96 years. Physical disability was present in 24% patients. There was statistically significant prevalence of psychological disorders in cases with physical impairment than those without physical impairment p value 0.001. Psychological disorders in stroke survivors are associated with physical disability in survivors of acute ischemic stroke. Present study also found association of diabetes with physical disability in cases of survivors of acute ischemic stroke.


Design, implementation, and evaluation of a novel curriculum to teach transurethral resection of the prostate (TURP): a 3-year experience of urology simulation bootcamp course.
To present the three-year experience of the multi-component TURP module at Urology Simulation Bootcamp Course (USBC) and demonstrate trainee's competence progression and satisfaction. During the USBC, a 4-h TURP module was developed and consisted of (a) familiarisation and assembly of resectoscope instrument, (b) didactic lecture on TURP operative techniques and postoperative complications, (c) learning hands-on resection on validated simulators [Samed, GmBH, Dresden, Germany; TURP Mentor™, Simbionix, Israel], and (d) practicing clot evacuation using the Ellik bladder Evacuator. Trainee's level of instrument knowledge, operative competence, and confidence were assessed pre- and post-course. Trainee's feedback was also collected. One hundred thirty trainees participated in the USBC between 2016 and 2018. Eighty-seven percent of trainees scored themselves as 1-3 (low confidence in resection) on a 5-point Likert scale. Trainees significantly improved in their ability to perform resectoscope assembly for resection, coagulation and incision by 33.6% (p < 0.001), 28.1% (p < 0.001) and 34.0% (p < 0.001), respectively. There was a significant improvement in scores in itemised technical skill on the TURP simulator following completion of the TURP module (Mean difference = 3.4 points, 95% CI 2-4, p < 0.001). Ninety-one percent of trainees agreed that the TURP module was useful for their development in urological training. Our results demonstrated that it is feasible to develop and implement a focussed module for teaching TURP with significant improvement in learning. Trainee feedback suggests that they were highly satisfied with the teaching provided and models used. This style of training can be implemented for other common surgical procedures.


Specificity and distribution of alpha antigens of Mycobacterium avium-intracellulare, Mycobacterium scrofulaceum, and related species of mycobacteria.
Specificity of antigenic determinants in alpha antigens of Mycobacterium avium, M. intracellulare, M. scrofulaceum, M. gordonae, and M. szulgai was investigated by the agar gel diffusion technique using the respective absorbed antialpha serum on a total of 225 strains classified into 13 species of slowly growing mycobacteria. The specific antigenic determinants in alpha antigen of M. scrofulaceum, M. gordonae, and M. szulgai were species specific, whereas those of M. avium and M. intracellulare were M. avium-M. intracellulare complex specific. Alpha antigens were found to be stable. These results suggest that the absorbed serums of antialpha- M. avium-intracellulare, -M. scrofulaceum, -M. gordonae, and -M. szulgai are useful for serologic identification of these species of slowly growing mycobacteria.