An In Vitro Evaluation of the Red Cell Damage and Hemocompatibility of Different Central Venous Catheters.
The aim of our study was to evaluate the damaging impact of characteristics of the central venous catheters (CVCs) on red blood cells. CVCs from three different manufacturers were analyzed, including the presence of coating, tunnel geometry, length, lumen diameter, and number of lumens with two respective flow rates (33 mL/min and 500 mL/min). Blood cell damage was defined by analyzing microparticle (MP) and hematologic analysis. MPs were isolated by ultracentrifugation of erythrocyte concentrate and analyzed on a flow cytometer. Characteristics of catheters were not associated with blood cell damage at a low flow rate but showed an effect with a high flow rate. CVCs with a polyhexanide methacrylate coating have caused statistically less blood cell damage than noncoated CVCs. The length of lumens, diameter, and geometry of the tunnel showed no differences in blood cell damage. Meanwhile, the number of lumens was predicted to have a greater effect on the erythrocyte damage, which was revealed with the formation of MPs and hematological parameters. CVCs with five lumens caused significantly less damage to the blood cells than CVCs with a single lumen. Moreover, a high flow rate of 500 mL/min caused less damage to the blood cells than a low rate of 33 mL/min. Properties of CVCs are an important factor for quality patient care, especially when transfusing blood with high flow rates, as we want to provide a patient with high-quality blood with as few damaged cells as possible.


Pre-clinical evaluation of cysteamine bitartrate as a therapeutic agent for mitochondrial respiratory chain disease.
Cysteamine bitartrate is a US Food and Drug Administration-approved therapy for nephropathic cystinosis also postulated to enhance glutathione biosynthesis. We hypothesized this antioxidant effect may reduce oxidative stress in primary mitochondrial respiratory chain (RC) disease, improving cellular viability and organismal health. Here, we systematically evaluated the therapeutic potential of cysteamine bitartrate in RC disease models spanning three evolutionarily distinct species. These pre-clinical studies demonstrated the narrow therapeutic window of cysteamine bitartrate, with toxicity at millimolar levels directly correlating with marked induction of hydrogen peroxide production. Micromolar range cysteamine bitartrate treatment in Caenorhabditis elegans gas-1(fc21) RC complex I (NDUFS2-/-) disease invertebrate worms significantly improved mitochondrial membrane potential and oxidative stress, with corresponding modest improvement in fecundity but not lifespan. At 10 to 100 μm concentrations, cysteamine bitartrate improved multiple RC complex disease FBXL4 human fibroblast survival, and protected both complex I (rotenone) and complex IV (azide) Danio rerio vertebrate zebrafish disease models from brain death. Mechanistic profiling of cysteamine bitartrate effects showed it increases aspartate levels and flux, without increasing total glutathione levels. Transcriptional normalization of broadly dysregulated intermediary metabolic, glutathione, cell defense, DNA, and immune pathways was greater in RC disease human cells than in C. elegans, with similar rescue in both models of downregulated ribosomal and proteasomal pathway expression. Overall, these data suggest cysteamine bitartrate may hold therapeutic potential in RC disease, although not through obvious modulation of total glutathione levels. Careful consideration is required to determine safe and effective cysteamine bitartrate concentrations to further evaluate in clinical trials of human subjects with primary mitochondrial RC disease.


Cerebrovascular Risk in Restless Legs Syndrome: Intima-Media Thickness and Cerebral Vasomotor Reactivity: A Case-Control Study.
Although some studies have suggested an association between cardiovascular disease and restless legs syndrome (RLS), the mechanisms underlying this relationship remain unclear. The intima-media thickness (IMT) and vasomotor reactivity are two simple, non-invasive tools to investigate preclinical atherosclerosis and microangiopathy, respectively. The aims of this study were to evaluate carotid IMT and to explore vasomotor reactivity in idiopathic RLS (iRLS) patients. We enrolled 44 iRLS after exclusion of patients with secondary causes of RLS, history of vascular events, known uncontrolled vascular risk factors and other neurological disorders. Forty-four age and sex matched controls were therefore recruited. No significant differences in demographic data and vascular risk factors were found between the two groups. Carotid IMT was measured with a high-resolution B-mode ultrasound on the far-wall of common carotid artery, 10 mm and 30 mm to the carotid bulb. Vasomotor reactivity to hypo- and hypercapnia was assessed, by right middle cerebral artery transcranial Doppler, accordingly to the changes in peak systolic velocity, peak diastolic velocity and mean blood flow velocity. Mean IMT was significantly increased in patients with iRLS when measured immediately proximally to carotid bifurcation (0.73; sd=0.17), versus controls (0.65; sd=0.13); p=0.035. Patients showed higher cerebrovascular flow velocities (CBFVs) compared to controls. After multivariate analysis, age, hypertension and iRLS proved to be independent IMT predictors. Increased IMT and higher CBFVs in iRLS support the association of iRLS with vascular damage, possibly through enhanced atherogenesis and sympathetic hyperactivity. However, to clarify a causal relationship, further longitudinal assessment of these parameters is needed, trying to control all their physiological modifying factors.


A Multilevel Analysis of Neighborhood Socioeconomic Effect on Preterm Births in Georgia, USA.
This study estimates the neighborhood socioeconomic status (SES) effect on the risk of preterm birth (PTB) using multilevel regression (MLR) models. Birth data retrieved from year 2000 and 2010 Georgia Vital Records were linked to their respective census tracts. Principle component analysis (PCA) was performed on nine selected census variables and the first two principal components (Fac1 and Fac2) were used to represent the neighborhood-level SES in the MLR models. Two-level random intercept MLR models were specified using 122,744 and 112,578 live and singleton births at the individual level and 1613 and 1952 census tracts at the neighborhood level, for 2000 and 2010, respectively. After adjustment for individual level factors, Fac1, which represents disadvantaged SES, respectively generated an Odds Ratio of 1.056 (95% CI: 1.031-1.081) and 1.080 (95% CI: 1.056-1.105) for these two years, showing a modest but statistically significant effect on PTB. After adjusting for individual level factors and the census tract level factors, Intra-class correlation (ICC) was 1.2% and 1.4%, for year 2000 and 2010, respectively. The two IOR-80% intervals, 0.73-1.52 (year 2000) and 0.73-1.59 (year 2010) suggest large unexplained between census tract variation. The Median Odds Ratio (MOR) value of 1.21(year 2000) and 1.23 (year 2010) revealed that the un-modeled neighborhood effect was smaller than two individual-level predictor variables, race, and tobacco use but larger than the fixed effect of census tract-level predicting variable, Fac1 and all the other individual level factors. Overall, better census tract level SES was found to have a modest protective effect for PTB risk and the effects of the two examined years were similar. Large unexplained between census tract heterogeneity warrants more sophisticated MLR models to further investigate the PTB risk factors and their interactions at both individual and neighborhood levels.


Quantitation of endogenous thyroid hormone receptors alpha and beta during embryogenesis and metamorphosis in Xenopus laevis.
Greater than 90% of the endogenous thyroid hormone receptor proteins TR alpha and TR beta in tissues of Xenopus laevis comigrate with their respective in vitro synthesized counterparts, and these major components are not phosphorylated detectably. Maternally inherited TR alpha protein is stable through early embryogenesis during a time in which there is no detectable TR alpha mRNA synthesis. At stage 35 when TR alpha mRNA is first detectable, the inherited TR alpha protein is present at about 100 molecules/cell. TR alpha protein subsequently increases to levels of about 1500 and 6000 molecules/cell in tail and head regions, respectively, in stage 52 tadpoles. Even though TR alpha mRNA gradually increases during metamorphosis (from stage 52 to 62), TR alpha protein remains constant, suggesting strongly that post-transcriptional events control the ultimate levels of TR alpha protein. In contrast, there is no detectable TR beta protein (less than 100 molecules/cell) throughout embryogenesis until stage 52. Both TR beta mRNA and protein rise along with the increase in endogenous thyroid hormone, reaching a maximum at the climax of metamorphosis, when TR beta protein exceeds TR alpha protein in concentration. As with TR alpha protein, TR beta protein in tail is consistently about one-fourth that of TR beta protein in the head region. The number of TR alpha protein molecules in extracts of premetamorphic tadpoles and cultured cells grown in the absence of thyroid hormone fully accounts for all of the sites to which 125I-T3 bind. We interpret this to mean that TR alpha protein must be a necessary, if not sufficient, component in the pathway toward metamorphosis triggered by thyroid hormone and required for the phenomenon of competence in tissues and cells.


Change in lifestyle behaviors and medication use after a diagnosis of ductal carcinoma in situ.
Women with ductal carcinoma in situ (DCIS) of the breast represent a growing cancer survivor population with a diagnosis of uncertain malignant potential. These survivors face an absence of scientific guidelines regarding lifestyle changes that can help to prevent a breast cancer recurrence. In this first report from the Wisconsin In Situ Cohort (WISC) study, we examine how women are currently changing their lifestyle behaviors and medication use following a diagnosis of DCIS. At study entry (1997-2006), 1,959 subjects (78% of eligible) with DCIS were identified from the Wisconsin cancer registry and administered an interview assessing behaviors prior to diagnosis. Follow-up interviews were completed every 2 years after the initial interview, beginning in 2003 and continuing through 2006. After adjusting for age and calendar year, women were 2.2 kg (95% CI 1.4, 3.0) heavier, 35% (95% CI 20, 47) less likely to be a smoker, 19% (95% CI -1, 43) more likely to use non-steroidal anti-inflammatory drugs, and 57% (95% CI 26, 95) more likely to use antidepressants after a DCIS diagnosis compared to 1 year prior to diagnosis. Use of postmenopausal hormones decreased sharply (OR = 0.06; 95% CI 0.04, 0.09) following a DCIS diagnosis. These findings indicate that women make substantial changes in their behaviors after a DCIS diagnosis. This cohort will be further monitored to evaluate the association between these behaviors and health outcomes following DCIS.


Antibody response to pneumococcal capsular polysaccharides in children with acute otitis media.
To describe the antibody response to pneumococcal capsular polysaccharides in children <2 years of age with pneumococcal acute otitis media (AOM) caused by serotypes 6A, 6B, 11A, 14, 19F or 23F. These serotypes were commonly found in both nasopharyngeal carriage and AOM in children of the study population in Finland. Serum antibody concentrations to pneumococcal capsular polysaccharides of types 6B, 11A, 14, 19F and 23F were measured by enzyme immunoassay in acute and convalescent sera from children with AOM. Responses (at least 2-fold increase of antibody concentration) were relatively infrequent and varied with both the age of the child and the serotype of the Streptococcus pneumoniae isolated from the middle ear fluid. Children older than 12 months were more likely to have antibody responses than were younger children. Responses were seen only infrequently to types 6A, 6B or 19F (1 of 14, 1 of 9 and 2 of 25, respectively), more often to types 11A and 14 (2 of 8 and 3 of 8) and relatively frequently to type 23F (8 of 18). However, the convalescent antibody concentrations to type 23F were low and usually declined after the infection, whereas responders to 14 AOM had antibodies that persisted at a high concentration through the follow-up. The results emphasize the differences between Streptococcus pneumoniae serotypes in their immunogenicity and quantitative and qualitative differences of antibodies produced after infection.


Artificial urinary sphincter implantation using a bulbous urethral cuff: perioperative care.
Urinary incontinence is a common problem encountered by clinicians and nursing staff. An artificial genitourinary sphincter (AGUS) placed around the bulbar urethra or bladder neck has become one of the gold standards for treating urinary incontinence due to sphincteric incompetence. It is imperative not only for physicians but also the nursing staff to have a working knowledge of the mechanics, risks, benefits, and durability of the AGUS to provide care for those patients encountered with an AGUS.


Characterization of gonadotropin-releasing hormone (GnRH) binding to pituitary receptors in goldfish (Carassius auratus).
Goldfish pituitary gonadotropin-releasing hormone (GnRH) receptors were characterized by using a superagonist analog of teleost GnRH (tGnRH-A; [D-Arg6, Trp7, Leu8, Pro9-NHEt]-GnRH). Equilibrium binding of 125I-tGnRH-A to a goldfish pituitary membrane preparation was achieved after a 30-min incubation at 4 degrees C; binding was significantly reduced after increasing incubation temperature to 22 degrees C. Binding of the radioligand was a function of tissue concentration, with a linear correlation over the range of 0.5-2 pituitary per tube. Incubation of the pituitary membrane preparation with increasing concentrations of 125I-tGnRH-A indicated saturable binding at radioligand concentrations of 470 pM and above. The binding of 125I-tGnRH-A was found to be reversible after addition of the cold analog, and the dissociation curve could be resolved into two linear components; slower rates of dissociation of 125I-tGnRH-A were observed after the addition of excess unlabeled tGnRH than after the addition of tGnRH-A, indicating that the analog is more effective in displacing the label than the native peptide. Addition of the cold analog displaced bound 125I-GnRH-A, and Scatchard analysis suggested the presence of at least two classes of binding sites: a high-affinity/low-capacity site and a low-affinity/high-capacity site. Bound 125I-GnRH-A was displaced by tGnRH from both sites in parallel to that observed with tGnRH-A, indicating that both peptides bind to the same classes of binding sites; however, tGnRH-A had a greater affinity for the receptors than the native tGnRH. These results demonstrated the presence and provided characterization of GnRH receptors in goldfish pituitary.


Video-assisted repair of cervical lung hernia.
Lung hernia is an extremely rare condition and the treatments vary. We report a case of cervical lung hernia without any trauma. The patient underwent video-assisted repair with a satisfactory result.


[Urothelial carcinoma clear cell variant of the urinary bladder : a case report].
A 69-year-old woman who had been treated with hemodialysis for 16 months for chronic renal failure presented with a chief complaint of gross hematuria in March 2007. Cystoscopic examination revealed a non-papillary, pedunculated tumor located in the right wall of the urinary bladder. A transurethral resection of the bladder tumor (TURBT) was performed in September 2007. The pathological diagnosis was urothelial carcinoma clear cell variant. The patient is now apparently free of disease 20 months after TURBT. Urothelial carcinoma clear cell variant of the urinary bladder is a rare primary tumor, and this is the fourth case of urothelial carcinoma clear cell variant of the bladder to be reported.


[Autoimmune diseases in the context of pandemic COVID-19].
Patient suffering from autoimmune diseases (AID) typically have an increased risk of infection, which is attributed to the disease itself, but also to immunosuppressive drugs (IS) and comorbidities. During the current COVID-19 outbreak, the way to manage these diseases remains elusive. Limited data is currently available on AID and IS in the context of this new coronavirus infection. To date, there is no evidence to support an increase in complications of COVID-19 in these patients. In addition, certain drugs that are commonly used to treat AID could be part of the therapeutic arsenal used in COVID-19. The purpose of this article is to review the unique aspects of patients with AID during the COVID-19 outbreak.


A case of a thoracic mass negative on thoracentesis diagnosed by cryobiopsy from the visceral pleura.
Thoracoscopy under local anaesthesia is recommended for malignant tumours with negative pleural effusion cytology. Cryobiopsy from the visceral pleura by thoracoscopy under local anaesthesia can provide more diagnostic options for patients with thoracentesis-negative malignant effusions. Here we present the first case in which this technique was used. The patient had a pleural metastasis that could not be diagnosed even with rapid cytology of the parietal pleura biopsy. Indications, technical pitfalls, and safety tips are discussed.


Agonist-independent inactivation and agonist-induced desensitization of the G protein-activated K+ channel (GIRK) in Xenopus oocytes.
The G-protein-activated K+ channels of the GIRK (Kir 3) family are activated by Gbetagamma subunits of heterotrimeric Gi/Go proteins. Atrial GIRK currents evoked by acetylcholine (ACh)1 via muscarinic m2 receptors (m2R) display prominent desensitization. We studied desensitization of basal and ACh-evoked whole-cell GIRK currents in Xenopus oocytes. In the absence of receptor and/or agonist, the basal GIRK activity showed inactivation which was prominent when the preparation was bathed in a low-Na+, high-K+ extracellular solution (96 mM [K+]out and 2 mM [Na+]out) but did not occur in a normal physiological solution. Ion substitution experiments showed that this basal, agonist-independent inactivation was caused by the decrease in [Na+]out rather than by the increased [K+]out. We hypothesize that it reflects a depletion of intracellular Na+. ACh-evoked GIRK currents desensitized faster than the basal ones. The agonist-induced desensitization was present when the preparation was bathed in all solutions tested, independently of [Na+]out. A protein kinase C (PKC) activator inhibited the GIRK currents both in high and low [Na+]out, apparently mimicking agonist-induced desensitization; however, a potent serine/threonine protein kinase blocker, staurosporine, blocked only a minor part of desensitization. We conclude that basal inactivation and agonist-induced desensitization are separate processes, the former caused by changes in Na+ concentrations, and the latter by unknown factor(s) with only a minor contribution of PKC.


Field evaluation of bednets impregnated with deltamethrin for malaria control.
Trials were undertaken in a hypoendemic area of malaria in an area bordering Vietnam, in Napo County of Guangxi Zhuang Autonomous Region, China. The aim was to compare the relative cost effectiveness of DDT residual spraying and of bednets impregnated with deltamethrin in the malaria control program. The trials were divided into three subgroups: (1) two farming areas and one coal mining area with a total population > 20,000, where the trial consisted of mass bednets impregnated with deltamethrin 15 mg/m2 net surface once a year, (2) one farming area with a population of approximately 3,600 where DDT residual spraying at 2g/m2 was carried out twice a year in May and August; (3) one farming area and one coal mining village with a population of > 4,000 were used as a control. The malaria vector population consisted mainly of Anopheles minimus and An. anthropophagus with a small contribution from An. sinensis. After bednets were impregnated with deltamethrin the mosquitos resting on the surface of the bednets decreased significantly, although there was less effect on the total vector population. The results showed that malaria incidence decreased significantly both in areas where impregnated bednets were used and in areas where residual spraying was undertaken. The positive IFAT rates of residents who slept under impregnated bednets decreased significantly in farming areas, especially in that area where bednet impregnation as a vector control measure had been undertaken for two years, but there was no change in the IFAT rate in DDT sprayed or control areas.(ABSTRACT TRUNCATED AT 250 WORDS)


The value of the white precursor cell channel (WPC) on the Sysmex XN-1000 analyser in a specialist paediatric hospital.
Historically, haematology analyser flags for abnormal white blood cells (WBCs) show good sensitivity but lower specificity, causing unnecessary blood film reviews. While the WBC differential channel on Sysmex XE and XN instruments reports a combined flag for blasts/abnormal lymphocytes, the new white precursor cell channel (WPC) on the XN series has been introduced to separate this into a specific flag for either cell type or, if no abnormality, remove the flag entirely. To compare the efficiency of abnormal WBC flags from the XN WPC to our existing analyser and determine whether WPC can reduce false positive flags and blood films required. Abnormal WBC flags from the Sysmex XE-5000 and XN-1000 were compared to manual differential and blood film morphology on 300 K2EDTA samples from infants and children. The XN WPC flag for blasts was more sensitive and specific than flags indicating blasts on the XE-5000, with a reduction in false positives from 64% (XE) to 36% (XN). Overall efficiency of the WPC flag for abnormal lymphocytes was 94% vs 79% on the XE. WPC reduced false positive flags for blasts and abnormal lymphocytes on neonatal samples by 50%. Automatic reflex analysis by WPC correctly removed a false positive flag from the white cell differential channel on 46% of samples. Total abnormal WBC flags from XN WPC were less (73) than the XE-5000 (92). XN WPC demonstrated superior efficiency of abnormal WBC flags on paediatric samples, compared to the XE-5000, with greater sensitivity and specificity of flagging, reducing blood films for review.


The effects of 10 weeks of β-alanine supplementation on peak power, power drop, and lactate response in Korean national team boxers.
This study was designed to investigate the effects of β-alanine (BA) supplementation on peak power, power drop, and lactate response in elite male amateur boxers. Nineteen male Korean national team boxers were divided into groups with either BA (n=9) or placebo (PL, n=10) supplementation. BA consumed 4.9-5.4 g/day of BA with training for 10 weeks and PL took PL in a similar manner. Physical fitness and lactate changes in sparring were measured before and after the 10-week intervention. Significant interactions (P<0.05) were shown for lower body peak power (P=0.049) and upper body power drop (P=0.042). Positive effects for the BA group were shown for lower body peak power (Cohen d=0.72; 95% confidence interval [CI], 0.09-1.35) and the maintenance of upper body power output (d=-0.91; 95% CI, -1.61 to -0.17). These findings suggest that Korean national amateur boxers who consumed BA demonstrated differential responses following a training intervention in specific physical fitness when compared to boxing athletes who consumed a PL.


Fibromyalgia in Older Individuals.
Fibromyalgia (FM) is a condition of chronic widespread pain (CWP) that can occur throughout the life cycle and is likely underrecognized in older patients. FM is associated with considerable suffering and reduction in quality of life and may occur as a unique condition, but in older patients is most likely to be associated with another medical illness. Understood mechanistically to be a sensitization of the nervous system, recently identified as nociplastic pain, FM is accepted as a valid medical illness that requires a positive diagnosis and directed treatments. The cornerstone of treatments for FM are nonpharmacologic interventions, with the understanding that medications provide only modest benefit for most patients, and with particular concern about adverse effects in older patients. If FM is not recognized, treatments may be misdirected to the other medical condition, with failure to address FM symptoms, leading to overall poor outcome. In contrast, new complaints in older patients should not immediately be attributed to FM, and physicians should be vigilant to ensure that onset of a new illness is not ignored. As FM is most often a lifelong condition, patients should be encouraged to identify their own personal strategies that can attenuate symptoms, especially when symptoms flare. Continued life participation should be the outcome goal.


Chromatographic analysis of polysaccharides extracted from Chinese Indocalamus tesselatus.
Three fractions of polysaccharides that have high biological activity were extracted from Chinese bamboo leaves (Indocalamus Tesselatus) with 0.9% sodium chloride, 1% ammonium oxalate and 85% ethanol. Their component sugars were investigated by paper chromatography developed with n-butanol-pyridine-water (6:4:3 by vol), and by gas chromatography with crown ester stationary-phase-fused silica capillary column. Their monosaccharide compositions are different from each other with glucuronic acid, galactose, arabinose, mannose, glucose, and xylose (52, 18, 9, 7 and 6%, respectively) in one fraction, only glucuronic acid in another and glucuronic acid, fucose, arabinose and xylose (36, 31, 24 and 9%, respectively) in the third fraction. The protection effect of two polysaccharides against experimental hepatitis was studied in mice.


Isolation of H and Leb active human salivary mucins and structures of their sugar chains with the H and Ley (Y) determinants.
Blood-group active mucins were prepared from an OLeb donor's saliva. Salivary proteins were carboxamidomethylated and fractioned on Sepharose CL-4B in the presence of sodium dodecyl sulfate. H and Leb active mucins were eluted at the void volume and were purified by Sepharose CL-2B chromatography. The purified mucins comprised 71% carbohydrate, 26% protein and 3.1% sulfate (by weight). Sugar chains of the mucins were released by alkaline borohydride treatment and fractionated by serial liquid chromatography. Three neutral oligosaccharides were isolated and submitted to sugar analysis and 1H-nuclear magnetic resonance spectroscopy. The following structures are proposed: Fuc alpha 1-2Gal beta 1-3[Fuc alpha 1-2Gal beta 1-4GlcNAc beta 1-6]GalNAcol, Fuc alpha 1-2Gal beta 1-3[Fuc alpha 1-2Gal beta 1-4[Fuc alpha 1-3]GlcNAc beta 1-6]GalNAcol and Fuc alpha 1-2Gal beta 1-3GlcNAc beta 1-3GalNAcol, which contain the H type 2, Ley (Y) and H type 1 determinants, respectively. The present report provides the first identification of some antigenic structures in human blood-group active salivary mucins.


Induction of Sp1 in differentiating human embryonal carcinoma cells triggers transcription of the fibronectin gene.
Cells of the human embryonal carcinoma line NEC14 proliferate as densely packed clusters consisting of small, polygonal stem cells and do not express a detectable level of fibronectin (FN). Upon induction of differentiation by treatment with N,N'-hexamethylene bisacetamide (HMBA), the level of FN mRNA increased steeply within 24 h and FN began to be accumulated, along with the organization of actin filaments in the cells. The FN promoter elements required for the activation were analyzed in reference to a cluster of GC boxes by using the chloramphenicol acetyltransferase (CAT) gene fused to 5' sequential-deletion derivatives of the promoter and promoters carrying base substitutions in the GC boxes. Among four GC boxes, GC boxes 2 and 3 had the greatest effect on promoter activation, and base substitutions in these GC boxes resulted in 80% reduction in promoter activity. The pattern of DNA-protein complex formation with these GC boxes changed drastically after induction of differentiation. The extract prepared from undifferentiated NEC14 cells formed fast-migrating complexes (UnD complexes), while the extract prepared from NEC14 cells treated with HMBA for 24 h formed slow-migrating complexes containing Sp1. Both complexes were formed predominantly with GC box 2. Base substitutions within the GC boxes completely abolished the formation of both UnD and Sp1 complexes. Consistent with these changes, the Sp1 level increased steeply within 24 h. Induction of Sp1 expression in NEC14 cells effectively stimulated the promoter activity of the transfected FN promoter-CAT constructs. These results indicate that activation of the FN promoter in differentiating NEC14 cells occurs by the steep induction of Sp1, which prevents an undifferentiated cell factor from binding to the Sp1 sites.


Iodine prophylaxis in the aftermath of the Chernobyl accident in the area of Sejny in north-eastern Poland.
The aim of our study was the estimation of the effects, possible side-effects and immunological reactions after the mass iodine prophylaxis following the Chernobyl nuclear disaster. 1457 subjects, aged 6-55 yrs, filled in our questionnaires and in 1191 of them the titres of antithyroid antibodies (TA) including ATMA--Anti-Human Thyroid Membrane Antibodies and TGAb--Anti-Thyreoglobulin Antibodies were estimated. Our study revealed that the prophylaxis, recommended in Poland only for children and teenagers was widely used (more than 90%) and relatively safe. No serious side effects, especially in pregnant women consuming KI were reported. The frequency and mean titres of TA were similar in adults who took and did not take stable iodine. The incidence of TA could be connected with many different factors including iodine deficiency, endemy of goitre as well as iodine prophylaxis. The reason of this phenomenon may be explained in long-term population studies.


Involvement of the cocaine-amphetamine regulated transcript peptide (CART 55-102) in the modulation of rat immune cell activity.
Cocaine-amphetamine regulated transcript peptides (CART) belong to a neuropeptide family expressed in the central nervous system, especially in the hypothalamus, and also in peripheral tissues. The physiological functions of CART include modulation of pituitary hormone release, regulation of body weight, and the control of feeding behavior and metabolic activity. The reciprocal relationships between CART and immune system function have to be established. Therefore, in the present study we aimed to investigate the influence of CART, administered intracerebroventricularly (icv), on selected immune parameters and pituitary-adrenal axis hormone secretion in the rat. In rats submitted to icv infusion of CART or artificial cerebrospinal fluid (aCSF, control) selected immune parameters: splenocyte proliferation (spontaneous and mitogen-stimulated) and peritoneal leukocyte (PTL) activity (spontaneous and phorbol myristate acetate (PMA)-stimulated) were examined 60 and 120 min after treatment. The direct effect of CART on splenocytes in culture in vitro was also examined. Concentration of adrenocorticotrophic hormone (ACTH) and corticosterone was also measured in serum of control and CART infused rats. Splenocytes isolated 60 min after CART infusion exhibited a decreased, albeit non-significant, ability to proliferate spontaneously and were unable to answering to the mitogenic stimulation. This effect was not seen 120 min after CART treatment, which restored splenocyte proliferation decreased by aCSF infusion. CART addition in vitro did not influence proliferation of splenocytes from control rats. Spontaneous activity of peritoneal leukocytes was not modified by CART infusion. PMA-stimulated PTL activity was significantly decreased in aCSF-infused rats 120 min after treatment and CART infusion antagonized this effect. Non-significant increase in serum cortisol after 60 min followed by a significant decrease after 120 min with no change in ACTH concentration was found. The immunomodulatory activity of icv-infused CART appears to consist in the creation of a short-lasting immunosuppressive internal milieu, followed by the immunostimulatory one. This first effect was most probably due to the activation of the HPA axis and/or other immunosuppressive peptides, but not through the direct action of CART on immune cells. Thus, CART appears to be short-lasting and indirect modulator of immunity.


Bedside diagnostics in dermatology: Viral, bacterial, and fungal infections.
Viral, bacterial, and fungal infections are frequently encountered in clinical practice, resulting in numerous cutaneous manifestations. Although diagnosis of these infections has changed over time because of technological advancements, such as polymerase chain reaction, bedside diagnostic techniques still play an important role in diagnosis and management, enabling rapid and low-cost diagnosis and implementation of appropriate therapies. This 2-part article will review both common and infrequent uses of bedside diagnostic techniques that dermatologists can incorporate into daily practice. This article examines the utility of bedside tests for the diagnosis of viral, bacterial, and fungal infections. The second article in this series reviews the use of bedside diagnostics for parasitic and noninfectious disorders.


Pyroptotic cells externalize eat-me and release find-me signals and are efficiently engulfed by macrophages.
Pathogenic intracellular bacteria often hijack macrophages for their propagation. The infected macrophages release IL-1β and IL-18 and simultaneously commit suicide, which is called pyroptosis; both responses require caspase-1. Here, we found that pyroptotic cells induced by microbial infection were efficiently engulfed by human monocytic THP-1-cell-derived macrophages or mouse peritoneal macrophages. This engulfment was inhibited by the D89E mutant of milk fat globule (MFG) epidermal growth factor (EGF) factor 8 (MFG-E8; a phosphatidylserine-binding protein) that has been shown previously to inhibit phosphatidylserine-dependent engulfment of apoptotic cells by macrophages, suggesting that the engulfment of pyroptotic cells by macrophages was also phosphatidylserine dependent. Using a pair of cell lines that respectively exhibited pyroptosis or apoptosis after muramyl dipeptide treatment, we showed that both pyroptotic and apoptotic cells bound to a T-cell immunoglobulin and mucin domain-containing 4 (Tim4; another phosphatidylserine-binding protein)-coated plate, whereas heat-killed necrotic cells did not, indicating that phosphatidylserine was externalized in pyroptosis and apoptosis but not in accidental necrosis. Macrophages engulfed apoptotic cells most efficiently, followed by pyroptotic and then heat-killed necrotic cells. Pyroptotic cells also released a macrophage attractant(s), 'find-me' signal, whose activity was diminished by apyrase that degrades nucleoside triphosphate to nucleoside monophosphate. Heat-killed necrotic cells and pyroptotic cells released ATP much more efficiently than apoptotic cells. These results suggest that pyroptotic cells, like apoptotic cells, actively induce phagocytosis by macrophages using 'eat-me' and find-me signals. Based on these results, a possible role of coordinated induction of pyroptosis and inflammatory cytokine production is discussed.


Microencapsulation improves inhibitory effects of transplanted olfactory ensheathing cells on pain after sciatic nerve injury.
Olfactory bulb tissue transplantation inhibits P2X2/3 receptor-mediated neuropathic pain. However, the olfactory bulb has a complex cellular composition, and the mechanism underlying the action of purified transplanted olfactory ensheathing cells (OECs) remains unclear. In the present study, we microencapsulated OECs in alginic acid, and transplanted free and microencapsulated OECs into the region surrounding the injured sciatic nerve in rat models of chronic constriction injury. We assessed mechanical nociception in the rat models 7 and 14 days after surgery by measuring paw withdrawal threshold, and examined P2X2/3 receptor expression in L4-5 dorsal root ganglia using immunohistochemistry. Rats that received free and microencapsulated OEC transplants showed greater withdrawal thresholds than untreated model rats, and weaker P2X2/3 receptor immunoreactivity in dorsal root ganglia. At 14 days, paw withdrawal threshold was much higher in the microencapsulated OEC-treated animals. Our results confirm that microencapsulated OEC transplantation suppresses P2X2/3 receptor expression in L4-5 dorsal root ganglia in rat models of neuropathic pain and reduces allodynia, and also suggest that transplantation of microencapsulated OECs is more effective than transplantation of free OECs for the treatment of neuropathic pain.


The HLA system in primary open angle glaucoma and in patients with pseudoexfoliation of the lens capsule (exfoliation or fibrillopathia epitheliocapsularis).
HLA typing of eighteen antigens in 43 patients with pseudoexfoliation of the lens capsule, 45 patients with capsular glaucoma, and 43 patients with primary open angle glaucoma was performed. No significant deviation from a control series of 45 subjects was found.


Modification of the cholesterol efflux properties of human serum by enrichment with phospholipid.
To investigate the importance of phospholipid in promoting cholesterol efflux from cells, phospholipid multilamellar vesicles were incubated with normal human serum and the efflux ability of these lipid-modified sera was tested. When incubated under appropriate conditions, both dimyristoylphosphatidylcholine (DMPC) and bovine brain sphingomyelin (BBSM) were shown to combine with components of human serum to form new protein:lipid complexes and to markedly enhance the ability of serum to promote efflux of cholesterol from Fu5AH cells. In particular, the high density lipoprotein (HDL) particles were altered in their composition and electrophoretic properties and the alpha-migrating species, which were reactive with antibodies to apo-A-I, were converted to larger, pre-beta-migrating particles, similar in electrophoretic properties to pre beta(2)-HDL. DMPC, but not BBSM, also generated particles with mobility similar to pre beta(2)-HDL; These species were demonstrably different from the discoidal complexes formed by reaction of DMPC with purified apoA-I. However, no change in cholesterol efflux potential was observed when serum was mixed with phospholipids that failed to interact or when cells were incubated with phospholipid multilamellar vesicles alone. To further identify the components of serum that become altered in their efflux potential after reaction with phospholipid, isolated lipoprotein fractions were incubated with DMPC or BBSM and it was found that only interaction with HDL caused enhancement of cholesterol efflux. In summary, cholesterol removal from the Fu5AH cells by serum can be promoted by adding phospholipid under conditions where new HDL-like complexes can be formed between the phospholipid and serum components, most notably apolipoprotein A-I.


[Immune reactions of the conjunctiva and cornea following severe chemical burns of the outer eye].
Conjunctival and corneal tissue samples from nine patients with severe changes after acid and alkali burns were examined immunohistochemically. Epithelial HLA Class II antigen expression was present in three of five conjunctival samples, and all of the antigen markers examined in the conjunctival stroma could be stained. HLA Class II expression was demonstrated in four out of five corneal tissue samples. The results show that in Grade III or IV chemical burns, with the diverse changes of the external eye that they produce, immunological and possibly also autoimmune mechanisms can influence the clinical course.


Androgens and male fertility.
Androgens play a crucial role in the development of male reproductive organs such as the epididymis, vas deferens, seminal vesicle, prostate and the penis. Furthermore, androgens are needed for puberty, male fertility and male sexual function. High levels of intratesticular testosterone, secreted by the leydig cells, are necessary for spermatogenesis. Intratesticular testosterone is mainly bound to androgen binding protein and secreted into the seminiferous tubules. Inside the sertoli cells, testosterone is selectively bound to the androgen receptor and activation of the receptor will result in initiation and maintenance of the spermatogenic process and inhibition of germ cell apoptosis. The androgen receptor is found in all male reproductive organs and can be stimulated by either testosterone or its more potential metabolite dihydrotestosterone. Severe defects of the androgen receptor may result in abnormal male sexual development. More subtle modulations can be a potential cause of male infertility. Treatment of an infertile man with testosterone does improve spermatogenesis, since exogenous administrated testosterone and its metabolite estrogen will suppress both GnRH production by the hypothalamus and Luteinising hormone production by the pituitary gland and subsequently suppress testicular testosterone production. Also, high levels of testosterone are needed inside the testis and this can never be accomplished by oral or parenteral administration of androgens. Suppression of testosterone production by the leydig cells will result in a deficient spermatogenesis, as can be seen in men taking anabolic-androgenic steroids. Suppression of spermatogenesis by testosterone administration is also the basis for the development of a male contraceptive. During cytotoxic treatment or irradiation suppression of intratesticular testosterone production cells may prevent irreversible damage to the spermotogonial stem cells.


Improved flux limits for neutrinos with energies above 10(22) eV from observations with the Westerbork Synthesis Radio Telescope.
Particle cascades initiated by ultrahigh energy neutrinos in the lunar regolith will emit an electromagnetic pulse with a time duration of the order of nanoseconds through a process known as the Askaryan effect. It has been shown that in an observing window around 150 MHz there is a maximum chance for detecting this radiation with radio telescopes commonly used in astronomy. In 50 h of observation time with the Westerbork Synthesis Radio Telescope array we have set a new limit on the flux of neutrinos, summed over all flavors, with energies in excess of 4x10(22) eV.


Evaluation of reduced-tillering (tin) wheat lines in managed, terminal water deficit environments.
Small or shrivelled wheat kernels (screenings) that reduce crop value are commonly produced in terminal drought environments. The aim of this study was to establish whether the incorporation of the tiller inhibition (tin) gene would contribute to maintenance of kernel weight and reductions in screenings under terminal water deficit. Five Silverstar near-isogenic lines contrasting in high and low tiller potential and their recurrent Silverstar parent were established at two plant densities under managed terminal water deficit (mild and severe) and irrigated conditions. With irrigation (grain yield of 5.6 t ha(-1)), kernels of all lines weighed ~31 mg, with restricted-tillering (R-tin) lines producing an average 15% lower grain yield. Under both mild and severe terminal water deficit (4.1 t ha(-1) and 2.8 t ha(-1)), free-tillering lines had relatively high screenings ranging from 11.9% to 16.2%. Compared with free-tillering lines, R-tin lines maintained large kernel weight (~29 mg kernel(-1)) and had 29% and 51% fewer screenings under the two stresses, and a significantly greater (+11%) grain yield under mild stress. Higher kernel weights in tin lines were realized even with the greater kernel number per spike. The higher kernel weight of the R-tin lines under stress conditions was associated with greater anthesis biomass and increased stem water-soluble carbohydrates, ensuring more assimilate for later translocation to filling grain. The incorporation of the tin gene into genetic material adapted to the target environments provides scope for improvement in both grain yield and kernel weight, and a reduction in screenings in terminal water deficit environments.


A bipedal mammalian model for spinal cord injury research: The tammar wallaby.
Background: Most animal studies of spinal cord injury are conducted in quadrupeds, usually rodents. It is unclear to what extent functional results from such studies can be translated to bipedal species such as humans because bipedal and quadrupedal locomotion involve very different patterns of spinal control of muscle coordination. Bipedalism requires upright trunk stability and coordinated postural muscle control; it has been suggested that peripheral sensory input is less important in humans than quadrupeds for recovery of locomotion following spinal injury. Methods: We used an Australian macropod marsupial, the tammar wallaby (Macropuseugenii), because tammars exhibit an upright trunk posture, human-like alternating hindlimb movement when swimming and bipedal over-ground locomotion. Regulation of their muscle movements is more similar to humans than quadrupeds. At different postnatal (P) days (P7-60) tammars received a complete mid-thoracic spinal cord transection. Morphological repair, as well as functional use of hind limbs, was studied up to the time of their pouch exit. Results: Growth of axons across the lesion restored supraspinal innervation in animals injured up to 3 weeks of age but not in animals injured after 6 weeks of age. At initial pouch exit (P180), the young injured at P7-21 were able to hop on their hind limbs similar to age-matched controls and to swim albeit with a different stroke. Those animals injured at P40-45 appeared to be incapable of normal use of hind limbs even while still in the pouch. Conclusions: Data indicate that the characteristic over-ground locomotion of tammars provides a model in which regrowth of supraspinal connections across the site of injury can be studied in a bipedal animal. Forelimb weight-bearing motion and peripheral sensory input appear not to compensate for lack of hindlimb control, as occurs in quadrupeds. Tammars may be a more appropriate model for studies of therapeutic interventions relevant to humans.


ESTABLISHING GENETIC CORRELATIONS INVOLVING PARASITE VIRULENCE.
For many host-parasite interactions, virulence is necessarily affected by population densities, transmission biology of the parasite, and proliferation of the parasite at the expense of its host. Attempts to experimentally demonstrate genetic correlations involving virulence therefore need to employ protocols controlling for variation in the number of successful infections (i.e., the end-point of transmissibility). If protocols are not controlled, then correlations may be spurious, as appears to be the case in recent experimental studies by Ebert (1994) and Ebert and Magnin (1997). There is a need to explore the modes of the evolution of each of the many sequential steps in nonsymbiotic and symbiotic phases of host-parasite associations and the implication of such evolution for overall virulence. I argue that it is the interdependence of these sequential steps (and not overall virulence) that should be at the center of attempts to establish genetic correlations.


Maleic anhydride and acetylene plasma copolymer surfaces for SPR immunosensing.
We report on the successful application of carboxyl-rich plasma polymerized (PP) films as a matrix layer for bioreceptor immobilization in surface plasmon resonance (SPR) immunosensing. Composition and chemical properties of the carboxyl-rich PP films deposited from a mixture of maleic anhydride and acetylene were investigated. Changes in the films stored in air, water, and buffer were studied and the involved chemical changes were described. Performance in SPR immunosensing was evaluated on interactions of human serum albumin (HSA) with a specific monoclonal antibody. The comparison with the mixed self-assembled monolayer of mercaptoundecanoic acid and mercaptohexanol (MUA/MCH) and one of the most widely used surfaces for SPR, the 2D and 3D carboxymethylated dextran (CMD), was presented to show the efficacy of plasma polymerized matrix layers for biosensing. The PP film-based SPR immunosensor provided a similar detection limit of HSA (100 ng/mL) as MUA/MCH- (100 ng/mL) and 3D CMD (50 ng/mL)-based sensors. However, the response levels were about twice higher in case of the PP film-based immunosensor than in case of MUA/MCH-based alternative. The PP film surfaces had similar binding capacity towards antibody as the 3D CMD layers. The response of PP film-based sensor towards HSA was comparable to 3D CMD-based sensor up to 2.5 μg/mL. For the higher concentrations (> 10 μg/mL), the response of PP film-based immunosensor was lower due to inaccessibility of active sites of the immobilized antibody inside the flat PP film surface. We have demonstrated that due to its high stability and cost-effective straightforward preparation, the carboxyl-rich PP films represent an efficient alternative to self-assembled monolayers (SAM) and dextran-based layers in label-free immunosensing. Graphical abstract.


Transition of the pregnancy rate of bisected bovine embryos after co-transfer with trophoblastic vesicles prepared from in vivo-cultured in vitro-fertilized embryos.
Bisected bovine embryos were co-transferred with trophoblastic vesicles (TVs). These TVs were prepared by dissection of conceptuses that were collected by uterine flushing after culture for seven days in the uterus following transfer of embryos derived by in vitro fertilization (IVF). Pregnancy diagnoses were performed twice, between Day 26 and Day 43 and between Day 38 and Day 73 post-estrus by ultrasonography. The pregnancy rate was significantly increased at first pregnancy diagnosis when demi-embryos were transferred with TVs (66.7%, 16/24) compared with the control group (34.5%, 10/29) (P < 0.05). Three losses occurred in the co-transfer group between the first and second pregnancy diagnosis. The final pregnancy rates according to delivered offspring were 41.7% (10/24) and 27.6% (8/29), respectively. There were no statistically significant differences between the pregnant and non-pregnant groups with regard to the average diameter of the TVs measured before transfer at three points during the gestation period. The birth weight and gestation lengths of the offspring were almost the same for the co-transfer and control groups. In the co-transfer group, the genetic identities of calves from the separated embryos were not affected by the TVs, as confirmed by parental blood type testing. Delivered offspring in co-transferred groups showed normal morphology. In conclusion, the present study indicates that co-transfer of TVs prepared from conceptuses cultured in vivo following transfer of IVF embryos enhances the fertility of demi-embryos during the early stages of pregnancy, as has similarly been shown in previous research for those prepared from in vivo embryos.


Patient stories, narratives of resistance and the ethics of humane care: a la recherche du temps perdu.
Over the past 20 years, the study of patient-physician relationships and the discourse of clinical encounters has become a major area of inquiry. Interest in these topics is closely linked to an ethic of humane care, reflecting widespread criticism of a technocratic approach to clinical practice. A narrow framing of this approach led to the neglect of a category of patients' socially grounded accounts of their illness and treatment experiences referred to as 'narratives of resistance'. The problem addressed in this article is the lack of connection, in research and practice, between an approach grounded in a humane care ethic and an ethic of social justice that addresses the impact of social and economic inequalities on rates of illness and access to health care. The aim is to open up a critical discussion among researchers, health care educators, practitioners and patients about relationships between ethical standpoints, clinical practice and research.


Clinical utility of hypo- and hyperpigmentation of skin in diffuse cutaneous systemic sclerosis.
Cutaneous involvement is an early manifestation of systemic sclerosis (SSc). Localized areas of 'salt and pepper skin' (S&P) may develop. We hypothesize that S&P skin occurs frequently in diffuse cutaneous (dc) SSc which can be used in its early diagnosis and may correlate with joint contractures. Sixty-five patients were recruited for this study. The demographic profiles of SSc were ascertained from hospital records. These patients fulfilled the 2013 American College of Rheumatology/European League Against Rheumatism classification criteria. Patients were examined for skin pigmentary changes, modified Rodnan skin score (mRSS), telengiectasias, calcinosis, arthritis and joint contractures and pruritus. Sixty-five patients (59 female) were recruited with median age of 62.87 years. Forty-four had limited cutaneous SSc, 16 dcSSc, five had scleroderma overlap syndrome. Multivariate stepwise logistic regression indicated that mRSS severity and the presence of contractures were independently (P < 0.05) associated with dcSSc. The strong positive association between S&P and mRSS severity may explain the non-significance of S&P in this analysis. If mRSS severity is not included in the logistic regression analysis, the presence of contractures and S&P (odds ratio = 15.1) show significant (P < 0.01) independent associations with the dcSSc subtype. S&P skin and pruritus were similar in patients with Scl-70 and anti-RNA polymerase antibodies. Anti-centromere antibodies were negatively associated with S&P (χ2 = 7.89, P = 0.005). Our study demonstrates strong association of S&P skin with dcSSc (69%), increased risk of pruritus and contractures. Its presence can be used as another clinical tool to diagnose dcSSc in early stages. Observing for S&P skin changes does not require much training.


[Effect of exogenous administration of steroids during the 1st month of pregnancy in sows and gilts on certain characteristics of the litter].
Four trials were performed to evaluate the influence of i. m. application of progesterone and estrogens to 444 experimental and control sows and gilts, kept mostly under large-scale production conditions, during the first month of pregnancy, as exerted on the number of born piglets, live born piglets and on the average birth weight of the live born piglet in litter. In the first trial the experimental sows and gilts on the 16th and 17th days of gravidity were applied 25 mg of progesterone (P4) and 12.5 micrograms of estrone (E1), in the second trial in the same phase of gravidity 25 mg of P4 with 1.25 micrograms of estradiol-17 beta (E2) and in the third trial in the identical gestation phase only 12.5 micrograms of E2 per head/day. In the fourth trial, experimental animals were given 0.55 mg of P4 with 0.275 micrograms of E1 per one kg of live weight, for six subsequent days between the 25th and 30th day of gravidity. In all trials the control animals were administered, under the same conditions, only the vehicle. None of the performed trials proved a significant difference in the studied parameters of litters of experimental and control animals (P greater than 0.05). The application of 25 mg of P4 with 1.25 micrograms of E2 on the 16th and 17th days of gravidity per sow/day under the conditions of small-scale production with the extension of the evaluated criteria of litter quality had no significant influence (P greater than 0.5) on the above-mentioned indicators, nor on the average birth weight of the whole litter and on the average number and weight of piglets of experimental sows at weaning. These results document that the verified application of hormonal substances in the above-mentioned phases of the first month of sow and gilt gravidity do not represent, in view of the intensive reproduction of pigs, a suitable procedure.


Dual regulation of tumor necrosis factor-α on myosin light chain phosphorylation in vascular smooth muscle.
We previously demonstrated that inhibitor κB kinase 2 (IKK2) is a myosin light chain kinase (MLCK). In the present study, we assess whether the prototypical activator of IKK2 tumor necrosis factor-α (TNF-α) regulates the MLCK activity of IKK2 and thus MLC phosphorylation in vascular smooth muscle cells (VSMCs). Kinase activity assay revealed that TNF-α downregulated the MLCK activity of IKK2 in human VSMCs (HVSMCs). However, Western blot analysis did not demonstrate a significant effect of TNF-α on MLC phosphorylation in HVSMCs, and myograph analysis did not reveal a significant effect of TNF-α on the contraction of the aorta from Sprague-Dawley rats and C57Bl/6j mice, suggesting a dual regulation of MLC phosphorylation by TNF-α. Confirming this notion, TNF-α significantly increased MLC phosphorylation in IKK2(-/-) but not wild-type cells. Furthermore, our results show that TNF-α increased GTP-bound RhoA and MLC phosphatase subunit MYPT1 phosphorylation and markedly reduced MLC phosphorylation in the presence of Rho-kinase inhibitor Y-27632, suggesting that downregulation of MLCK activity of IKK2 by TNF-α is antagonized by simultaneous RhoA/Rho-kinase activation. These results indicate that TNF-α dually regulates MLC phosphorylation through both IKK2 and RhoA/Rho-kinase pathways.


Structural basis for binding of the plasmid ColIb-P9 antisense Inc RNA to its target RNA with the 5'-rUUGGCG-3' motif in the loop sequence.
The sequence 5'-rUUGGCG-3' is conserved within the loop regions of antisense RNAs or their targets involved in replication of various prokaryotic plasmids. In IncIalpha plasmid ColIb-P9, the partially base paired 21-nucleotide loop of a stem-loop called structure I within RepZ mRNA contains this hexanucleotide sequence, and comprises the target site for the antisense Inc RNA. In this report, we find that the base pairing interaction at the 5'-rGGC-3' sequence in the hexanucleotide motif is important for interaction between Inc RNA and structure I. In addition, the 21-base loop domain of structure I is folded tighter than predicted, with the hexanucleotide sequence at the top. The second U residue in the sequence is favored for Inc RNA binding in a base-specific manner. On the other hand, the upper domain of the Inc RNA stem-loop is loosely structured, and maintaining the loop sequence single-stranded is important for the intermolecular interaction. Based on these results, we propose that a structural feature in the loop I domain, conferred probably by the conserved 5'-rUUGGCG-3' sequence, favors binding to a complementary, single-stranded RNA. This model also explains how the RepZ mRNA pseudoknot, described in the accompanying paper (Asano, K., and Mizobuchi, K. (1998) J. Biol. Chem. 273, 11815-11825) is formed specifically with structure I. A possible conformation adopted by the 5'-rUUGGCG-3' loop sequence is discussed.


Accelerometer-Based Gait Recognition by Sparse Representation of Signature Points With Clusters.
Gait, as a promising biometric for recognizing human identities, can be nonintrusively captured as a series of acceleration signals using wearable or portable smart devices. It can be used for access control. Most existing methods on accelerometer-based gait recognition require explicit step-cycle detection, suffering from cycle detection failures and intercycle phase misalignment. We propose a novel algorithm that avoids both the above two problems. It makes use of a type of salient points termed signature points (SPs), and has three components: 1) a multiscale SP extraction method, including the localization and SP descriptors; 2) a sparse representation scheme for encoding newly emerged SPs with known ones in terms of their descriptors, where the phase propinquity of the SPs in a cluster is leveraged to ensure the physical meaningfulness of the codes; and 3) a classifier for the sparse-code collections associated with the SPs of a series. Experimental results on our publicly available dataset of 175 subjects showed that our algorithm outperformed existing methods, even if the step cycles were perfectly detected for them. When the accelerometers at five different body locations were used together, it achieved the rank-1 accuracy of 95.8% for identification, and the equal error rate of 2.2% for verification.


MetaCoMET: a web platform for discovery and visualization of the core microbiome.
A key component of the analysis of microbiome datasets is the identification of OTUs shared between multiple experimental conditions, commonly referred to as the core microbiome. We present a web platform named MetaCoMET that enables the discovery and visualization of the core microbiome and provides a comparison of the relative abundance and diversity patterns between subsets of samples within a microbiome dataset. MetaCoMET provides an efficient and interactive graphical interface for analyzing each subset defined by the union or disjunction of groups within the Venn diagram, and includes a graphical taxonomy summary, alpha diversity metrics, Principal Coordinate analysis, abundance-based heatmaps, and a chart indicating the geographic distribution of each sample. MetaCoMET is a user-friendly and efficient web platform freely accessible at http://probes.pw.usda.gov/MetaCoMET or http://aegilops.wheat.ucdavis.edu/MetaCoMET CONTACT: email@example.com information: Supplementary data are available at Bioinformatics online.


Identification of critical residues in the amino terminal domain of the human NR2B subunit involved in the RO 25-6981 binding pocket.
N-Methyl-d-aspartate (NMDA) receptors play key roles in both physiological processes, particularly synaptic plasticity, and in neuropathological states such as epilepsy and acute neurodegeneration. R-(R*,S*)-alpha-(4-Hydroxyphenyl)-beta-methyl-4-(phenyl-methyl)-1-piperidine propanol (RO 25-6981), is a high-affinity and selective blocker of NMDA receptors containing the NR2B subunit. Using site-directed mutagenesis, [3H]RO 25-6981 binding, Xenopus oocyte voltage-clamp recordings, and molecular modeling, we have identified several critical residues involved in the RO 25-6981 binding site within the N-terminal LIVBP-like domain of the human NR2B subunit. Two mutations, NR2B(D101A) and NR2B(F176A), resulted in a complete loss of [3H]RO 25-6981 binding and also abolished the high-affinity RO 25-6981-mediated inhibition of NMDA-induced currents. The mutation NR2B(T233A) led to a marked reduction in binding affinity by 13-fold. Mutations F182A, D104A, or K234A had a more moderate influence on the binding affinity (KD values increased by 8-, 7-, and 6-fold, respectively). In a three-dimensional model of the NR2B LIVBP-like domain based on the X-ray crystal structure of the amino-terminal domain of the mGlu1 receptor, the critical residues are located in the central cleft where interaction with RO 25-6981 may stabilize the closed structure of the domain. Our results suggest that the three amino acids Asp-101, Phe-176, and Thr-233 are important molecular determinants for the high-affinity binding of RO 25-6981 to the LIVBP-like domain of human NR2B. A possible binding mode for RO 25-6981 is proposed.


In vivo and in vitro surface changes in a highly cross-linked polyethylene.
Highly cross-linked, ultrahigh-molecular-weight polyethylenes (UHMWPEs) were developed to reduce UHMWPE wear in arthroplasty. These UHMWPEs have manifested an improvement in the wear resistance. Examination of the first retrievals revealed surface features not usually observed on conventional retrievals. A flattening of the machining marks is evident, together with the presence of ripples with microfissures. These ripples were investigated in vitro and on retrievals having a follow-up of up to 15 months. The examinations of all specimens showed that the ripples may be described as folds with microcracks. The depth of the microcracks extended to a maximum of 5 microm and is independent of the mode of loading (up to 27 million cycles). Because of the extreme wear resistance of these UHMWPEs, the folds accumulate on the surface of components.


Impact of kinship matrices on genetic gain and inbreeding with optimum contribution selection in a genomic dairy cattle breeding program.
Genomic selection has increased genetic gain in dairy cattle, but in some cases it has resulted in higher inbreeding rates. Therefore, there is need for research on efficient management of inbreeding in genomically-selected dairy cattle populations, especially for local breeds with a small population size. Optimum contribution selection (OCS) minimizes the increase in average kinship while it maximizes genetic gain. However, there is no consensus on how to construct the kinship matrix used for OCS and whether it should be based on pedigree or genomic information. VanRaden's method 1 (VR1) is a genomic relationship matrix in which centered genotype scores are scaled with the sum of 2p(1-p) where p is the reference allele frequency at each locus, and VanRaden's method 2 (VR2) scales each locus with 2p(1-p), thereby giving greater weight to loci with a low minor allele frequency. We compared the effects of nine kinship matrices on genetic gain, kinship, inbreeding, genetic diversity, and minor allele frequency when applying OCS in a simulated small dairy cattle population. We used VR1 and VR2, each using base animals, all genotyped animals, and the current generation of animals to compute reference allele frequencies. We also set the reference allele frequencies to 0.5 for VR1 and the pedigree-based relationship matrix. We constrained OCS to select a fixed number of sires per generation for all scenarios. Efficiency of the different matrices were compared by calculating the rate of genetic gain for a given rate of increase in average kinship. We found that: (i) genomic relationships were more efficient than pedigree-based relationships at managing inbreeding, (ii) reference allele frequencies computed from base animals were more efficient compared to reference allele frequencies computed from recent animals, and (iii) VR1 was slightly more efficient than VR2, but the difference was not statistically significant. Using genomic relationships for OCS realizes more genetic gain for a given amount of kinship and inbreeding than using pedigree relationships when the number of sires is fixed. For a small genomic dairy cattle breeding program, we recommend that the implementation of OCS uses VR1 with reference allele frequencies estimated either from base animals or old genotyped animals.


Ab initio phonon properties of half-Heusler NiTiSn, NiZrSn and NiHfSn.
A theoretical investigation of phonon properties from first-principles calculations is carried out for the half-Heusler compounds NiXSn, [Formula: see text], Zr and Hf. The crystal structures are optimised via ab initio calculations within the framework of density functional theory. The phonon properties are retrieved from harmonic and anharmonic interatomic force constants calculations using the finite size displacements method and many-body perturbation theory. A solution to the linearized phonon Boltzmann transport equation is then used to compute the ab initio thermal conductivities. For X = Ti, Zr and Hf, we found 15.4, 13.3 and 15.8 W m(-1) K(-1) at 300 K, respectively. Thanks to a spectral analysis of the velocities and lifetimes we were able appreciate the differences in the thermal conductivities between the three compounds under study. Our results provide insights to understand the behaviour of the thermal conductivity and therefore to improve the thermoelectric figure of merit for such materials.


Behavioral characteristics and neural mechanisms mediating performance in a rodent version of the Balloon Analog Risk Task.
The tendency for some individuals to partake in high-risk behaviors (eg, substance abuse, gambling, risky sexual activities) is a matter of great public health concern, yet the characteristics and neural bases of this vulnerability are largely unknown. Recent work shows that this susceptibility can be partially predicted by laboratory measures of reward seeking under risk, including the Balloon Analog Risk Task. Rats were trained to respond on two levers: one of which (the 'add lever') increased the size of a potential food reward and a second (the 'cash-out lever') that led to delivery of accrued reward. Crucially, each add-lever response was also associated with a risk that the trial would fail and no reward would be delivered. The relative probabilities that each add-lever press would lead to an addition food pellet or to trial failure (risk) were orthogonally varied. Rats exhibited a pattern of responding characteristic of incentive motivation and risk aversion, with a subset of rats showing traits of high-risk taking and/or suboptimal responding. Neural inactivation studies suggest that the orbitofrontal cortex supports greater reward seeking in the presence or absence of risk, whereas the medial prefrontal cortex is required for optimization of patterns of responding. These findings provide new information about the neural circuitry of decision making under risk and reveal new insights into the biological determinants of risk-taking behaviors that may be useful in developing biomarkers of vulnerability.


[Granulocyte-colony stimulating factor producing tumor with high serum interleukin-6].
A 53-year-old man admitted to our hospital because of fever and chest abnormal shadow. Chest X-ray and computed tomography (CT) scan revealed large tumor on right upper lobe. Serum interleukin (IL)-6 and granulocyte-colony stimulating factor (G-CSF) were high. Right upper lobectomy and chest wall resection was performed. Histological diagnosis was large cell carcinoma. Immunohistological examination of lung tumor cells showed positive staining for G-CSF in only 1% of them. We diagnosed that tumor was G-CSF producing tumor and we thought that tumor produced IL-6.


Theoretical studies on the structural and spectroscopic properties of an iminocoumarin-based probe and its metal complexation: an implication for a fluorescence probe.
To understand the sensing behaviors of molecular fluorescent probes, an N,N-di(picolyl)aminoethyl-iminocoumarin probe (L) and its complexation with metal(II) ions (ML, M = Mg, Ca, Zn, Cd and Hg) were examined by relativistic density functional theory (DFT). Four stable conformational isomers (labeled as g1, g2, a1 and a2) for each of them have been optimized, except for CaL having only three without the g2 isomer. All of these structures have been confirmed by frequency calculations. In the aqueous solution, the a2 isomer of the L probe was calculated to be the most stable, while the g1 isomer turns out to be energetically favorable upon binding with metal ions. At these isomeric geometries, the experimentally obtained absorption was well reproduced by calculations of time-dependent DFT (TD-DFT) and a conductor-like polarized continuum model (CPCM). A slight red-shifting from L (508 nm) to ML (516-528 nm) was found. This is due to the metal affinity that stabilizes the LUMOs of ML greater than the HOMOs. Singlet excited-state structures of L and ML (M = Zn, Cd and Hg) were fully optimized using the TD-DFT approach, giving more relaxed geometries than their respective ground-state ones. Their fluorescent emissions in the aqueous solution were calculated to be 543 and 551-560 nm, respectively, agreeing with experimental values of 543 nm for L and 558 nm for ZnL. The present study also presents theoretical support for a sensing mechanism of photo-induced charge transfer of the L probe that was proposed in the previous experiment.


Utilization of commercial laboratory results in management of hyperandrogenism in women.
To compare results from testosterone radioimmunoassay kits commonly used by commercial laboratories as well as their reference ranges and to analyze the scientific literature for ranges of serum testosterone levels in normal women and those with hyperandrogenism. We reviewed quality assurance reports of various testosterone ligand challenges from four groups of laboratories and summarized testosterone data from 17 published reports about normal women and 14 studies of hyperandrogenic women. A significant variability was demonstrated between the radioimmunoassay kits at all concentrations (for example, a sample with a mean testosterone level of 96.1 ng/dL was reported by some laboratories as containing 71.8 ng/dL and by others as 123.4 ng/dL). All laboratories provide essentially the same "reference range" (approximately 10 to 90 ng/dL) but do not report how the range was established. The scientific literature clearly shows a significant separation in serum testosterone levels between normal (that is, not hyperandrogenic) and hyperandrogenic women. Most hyperandrogenic women had testosterone levels >50 ng/dL, whereas most normal control subjects had levels <40 ng/dL. Thus, most of these women with hyperandrogenism would have been considered to have normal testosterone levels if the reference ranges of commercial laboratories were used. These data illustrate the difficulty that physicians face when they are required to use different commercial laboratories to measure serum testosterone levels. We propose that (1) reference ranges be established on a clinically defined population for each hormone and method used, (2) laboratory reports include information about method and reference range population, and (3) physicians be allowed to choose which laboratories are used for their patients' hormone determinations, for consistency of results.


Targeting glycosylation pathways and the cell cycle: sugar-dependent activity of butyrate-carbohydrate cancer prodrugs.
Short-chain fatty acid (SCFA)-carbohydrate hybrid molecules that target both histone deacetylation and glycosylation pathways to achieve sugar-dependent activity against cancer cells are described in this article. Specifically, n-butyrate esters of N-acetyl-D-mannosamine (But4ManNAc, 1) induced apoptosis, whereas corresponding N-acetyl-D-glucosamine (But4GlcNAc, 2), D-mannose (But5Man, 3), or glycerol (tributryin, 4) derivatives only provided transient cell cycle arrest. Western blots, reporter gene assays, and cell cycle analysis established that n-butyrate, when delivered to cells via any carbohydrate scaffold, functioned as a histone deacetylase inhibitor (HDACi), upregulated p21WAF1/Cip1 expression, and inhibited proliferation. However, only 1, a compound that primed sialic acid biosynthesis and modulated the expression of a different set of genes compared to 3, ultimately killed the cells. These results demonstrate that the biological activity of butyrate can be tuned by sugars to improve its anticancer properties.


Results of combination chemotherapy of non-Hodgkin's lymphoma.
The analysis of the results of CVP and MOPP chemotherapy in 80 patients with advanced stages of non-Hodgkin's lymphoma shows that 36 achieved a complete remission. Twenty-five percent of all patients remain free of disease for periods ranging from 4 months to over 7 years, with a projected median duration of complete remissions of 3 1/2 years. Well differentiated and nodular histology were positive determinants for survival, confirming the overall clinical validity of the Rappaport classification system for the non-Hodgkin's lymphomata. However, it has been demonstrated that it is possible to achieve a complete remission using combination chemotherapy even in the most clinically aggressive histological subgroups, and that these responses can be correlated with an extended survival. An analysis of patterns of relapse from complete remission in patients with non-Hodgkin's lymphoma treated with combination chemotherapy has demonstrated differences between histological subgroups. Patients with diffuse histiocytic lymphoma who achieved a complete remission after 6 months of treatment without maintenance have remained disease-free, whereas those with nodular poorly differentiated lymphocytic and diffuse well differentiated lymphocytic lymphomata demonstrate a pattern of continuous late recurrence. The initial sites of relapse from complete remission in lymphocytic lymphoma were lymph nodes and bone marrow which were involved before treatment.


Evidence of the late lignification of the G-layer in Simarouba tension wood, to assist understanding how non-G-layer species produce tensile stress.
To recover verticality after disturbance, angiosperm trees produce 'tension wood' allowing them to bend actively. The driving force of the tension has been shown to take place in the G-layer, a specific unlignified layer of the cell wall observed in most temperate species. However, in tropical rain forests, the G-layer is often absent and the mechanism generating the forces to reorient trees remains unclear. A study was carried out on tilted seedlings, saplings and adult Simarouba amara Aubl. trees-a species known to not produce a G-layer. Microscopic observations were done on sections of normal and tension wood after staining or observed under UV light to assess the presence/absence of lignin. We showed that S. amara produces a cell-wall layer with all of the characteristics typical of G-layers, but that this G-layer can be observed only as a temporary stage of the cell-wall development because it is masked by a late lignification. Being thin and lignified, tension wood fibres cannot be distinguished from normal wood fibres in the mature wood of adult trees. These observations indicate that the mechanism generating the high tensile stress in tension wood is likely to be the same as that in species with a typical G-layer and also in species where the G-layer cannot be observed in mature cells.


[The composition of ascites lymphocytes in high-grade serous ovarian cancer and its correlation with clinical features].
Objective: To explore the composition of ascites lymphocytes in high-grade serous ovarian carcinoma (HGSOC) and its correlation with clinical features. Methods: A total of 59 newly-diagnosed HGSOC patients, aged (58±11) years old, who were treated at the Department of Obstetrics and Gynecology of Peking University Third Hospital from July 2019 to December 2020 were included, collecting ascites and peritoneal irrigation fluid respectively. Detect the proportion of T, B, NK cell and its subpopulations by flow cytometry, and analyze its correlation with the clinical characteristics of patients. Results: Among 59 patients, 48 patients (81.4%) had ascites, and 11 patients (18.6%) had no ascites. Compared with the peritoneal irrigation fluid, the CD3+T (70.2%±15.6% vs 78.1%±6.7%, P=0.014), CD8+T (38.3%±11.2% vs 47.7%±10.1%, P=0.014) and CD16-CD56bright NK [2.0% (0.8%, 3.6%) vs 4.2% (1.5%, 7.1%), P=0.026] cells were significantly decreased in the ascites, while the CD16+CD56dim NK cells was significantly increased [6.8% (2.8%, 15.7%) vs 2.6% (1.6%, 4.3%), P=0.008]. In patients with ascites volume ≥1 000 ml, CD16-CD56bright NK cells were significantly increased than those ascites volume<1 000 ml [3.1% (1.2%, 3.9%) vs 0.8% (0.4%, 2.3%), P=0.002]. Age was significantly positively correlated with the level of CD4+T cells (r=0.332, P=0.021) and the ratio of CD4+T/CD8+T (r=0.379, P=0.008) in ascites. In ascites from patients with poor response to treatment, the proportion of CD4+T cells was increased significantly than those with better response (64.7%±4.2% vs 48.3%±11.7%, P=0.002). Conclusion: The formation of ascites in newly diagnosed high-grade serous epithelial ovarian cancer patients would affect the lymphocyte composition in the abdominal cavity microenvironment, which is related to the patient's age, cancer progression and treatment response.


Capnocytophaga: a review of the literature.
Capnocytophaga species are normal mouth flora but can be opportunistic pathogens causing juvenile peridontitis and bacteremia in the compromised host. Indole-negative fusiforms isolated anaerobically or in the presence of increased CO2 can presumptively be identified as Capnocytophaga species.


Iron Reduces M1 Macrophage Polarization in RAW264.7 Macrophages Associated with Inhibition of STAT1.
Iron metabolism in inflammation has been mostly characterized in macrophages exposed to pathogens or inflammatory conditions. The aim of this study is to investigate the cross-regulatory interactions between M1 macrophage polarization and iron metabolism. Firstly, we characterized the transcription of genes related to iron homeostasis in M1 RAW264.7 macrophages stimulated by IFN-γ. The molecular signature of M1 macrophages showed high levels of iron storage (ferritin), a low level of iron export (ferroportin), and changes of iron regulators (hepcidin and transferrin receptors), which favour iron sequestration in the reticuloendothelial system and are benefit for inflammatory disorders. Then, we evaluated the effect of iron on M1 macrophage polarization. Iron significantly reduced mRNA levels of IL-6, IL-1β, TNF-α, and iNOS produced by IFN-γ-polarized M1 macrophages. Immunofluorescence analysis showed that iron also reduced iNOS production. However, iron did not compromise but enhanced the ability of M1-polarized macrophages to phagocytose FITC-dextran. Moreover, we demonstrated that STAT1 inhibition was required for reduction of iNOS and M1-related cytokines production by the present of iron. Together, these findings indicated that iron decreased polarization of M1 macrophages and inhibited the production of the proinflammatory cytokines. The results expanded our knowledge about the role of iron in macrophage polarization.


Cryptococcus-Epithelial Interactions.
The fungal pathogen, Cryptococcus neoformans, causes devastating levels of morbidity and mortality. Infections with this fungus tend to be predominantly in immunocompromised individuals, such as those with HIV. Infections initiate with inhalation of cryptococcal cells and entry of the pathogen into the lungs. The bronchial epithelial cells of the upper airway and the alveolar epithelial cells of the lower airway are likely to be the first host cells that Cryptococcus engage with. Thus the interaction of cryptococci and the respiratory epithelia will be the focus of this review. C. neoformans has been shown to adhere to respiratory epithelial cells, although if the role of the capsule is in aiding or hindering this adhesion is debatable. The epithelia are also able to react to cryptococci with the release of cytokines and chemokines to start the immune response to this invading pathogen. The activity of surfactant components that line this mucosal barrier towards Cryptococcus and the metabolic and transcriptional reaction of cryptococci when encountering epithelial cells will also be discussed.


Lymphoepithelial cyst of the pancreas. Report of a case diagnosed by fine needle aspiration biopsy.
Lymphoepithelial cysts of the pancreas are very rare lesions. To the best of our knowledge, 24 cases are reported in the literature. A 64-year-old male presented with gastrointestinal complaints. A computed tomography scan of the abdomen disclosed a 55 x 30-mm, homogeneous mass anterior to and contiguous with the pancreas. Fine needle aspiration biopsy (FNAB) showed threads of tissue composed of stratified squamous epithelium with subepithelial lymphocytic infiltrate and keratinous material. This case illustrates that FNAB is a rapid and reliable technique that can be used as the first diagnostic step in cases of cystic lesions of the pancreas. To our knowledge, this is the third case reported of this entity diagnosed with FNAB.


Formalin can alter the intracellular localization of some transcription factors in Saccharomyces cerevisiae.
Indirect immunofluorescence (IF) microscopy is a frequently used method to determine intracellular protein localization. It is especially useful for low abundance proteins, for example the GATA-factors (Gln3, Gat1) which activate nitrogen catabolite repression (NCR)-sensitive transcription. Limiting nitrogen or treating cells with Tor pathway inhibitor, rapamycin, elicits nuclear GATA-factor localization and increased NCR-sensitive transcription, whereas excess nitrogen restricts these proteins to the cytoplasm and decreases transcription. The initial step of the IF procedure is formalin-fixation that quenches cellular activity and fixes protein locations via cross-linking. We find that under some conditions, formalin itself can influence GATA-factor localization. With low formalin (0.8% or 1.6%), Gat1-Myc(13) became more nuclear, and with higher concentrations (5.6%), it became more cytoplasmic. Gln3-Myc(13) localization, on the other hand, did not respond to low formalin, but became more cytoplasmic at the higher concentration. Interestingly, the high concentration of formalin had no demonstrable effect when the GATA factors were completely nuclear, i.e. after rapamycin (Gat1-Myc(13)) or Msx (Gln3-Myc(13)) treatment. These effects are most likely elicited by polyoxymethylene glycols, which significantly increase the osmolarity of the medium (0.5-2). We suggest that varying degrees of osmotic stress and transcription factor movement in response to it can occur after the beginning of fixation but before proteins become immobilized.


Re-innervation of fast and slow twitch muscle following nerve crush at birth.
1. The frequency of miniature end-plate potentials (m.e.p.p.s) was significantly greater in the fast twitch extensor digitorum longus muscle (extensor) than in the slow twitch soleus, even though end-plate surface area was greater for fibres in the latter muscle. 2. Crush of the sciatic nerve at birth did not prevent the appearance of this difference in m.e.p.p. frequency. However, the frequency of the potentials in the re-innervated muscles was less than normal, even though the regenerated neuromuscular junction was qualitatively normal in morphology. 3. Though the re-innevated muscles were differentiated with respect to twitch time course, the extensor muscle was more responsive than normal to the contracture-inducing action of caffeine. 4. The Z line of the re-innervated extensor muscle was similar to that of the normal soleus in thickness. 5. Resting potential, passive electrical properties and action potential generating mechanism of the sarcolemma were normal. 6. Since the re-innervated muscles lacked muscle spindles, a role of sensory feed-back in the function of the neuromuscular junction as well as the neutrotrophic regulation of muscle is discussed.


Effect of platelet lysate on human cells involved in different phases of wound healing.
Platelets are rich in mediators able to positively affect cell activity in wound healing. Aim of this study was to characterize the effect of different concentrations of human pooled allogeneic platelet lysate on human cells involved in the different phases of wound healing (inflammatory phase, angiogenesis, extracellular matrix secretion and epithelialization). Platelet lysate effect was studied on endothelial cells, monocytes, fibroblasts and keratinocytes, in terms of viability and proliferation, migration, angiogenesis, tissue repair pathway activation (ERK1/2) and inflammatory response evaluation (NFκB). Results were compared both with basal medium and with a positive control containing serum and growth factors. Platelet lysate induced viability and proliferation at the highest concentrations tested (10% and 20% v/v). Whereas both platelet lysate concentrations increased cell migration, only 20% platelet lysate was able to significantly promote angiogenic activity (p<0.05 vs. control), comparably to the positive control. Both platelet lysate concentrations activated important inflammatory pathways such as ERK1/2 and NFκB with the same early kinetics, whereas the effect was different for later time-points. These data suggest the possibility of using allogeneic platelet lysate as both an alternative to growth factors commonly used for cell culture and as a tool for clinical regenerative application for wound healing.


Personal exposure to ultrafine particles: Two-level statistical modeling of background exposure and time-activity patterns during three seasons.
Personal exposure to air pollution is associated with time- and location-specific factors including indoor and outdoor air pollution, meteorology and time activities. Our investigation aims at the description and identification of factors determining personal exposure to particle number concentration (PNC) in everyday situations. Ten volunteers recorded their personal exposure to PNC and kept an activity diary in three different seasons besides stationary measurements of ambient air pollution and meteorology. Background exposure to PNC was modelled using the most predictive variables. In a second step, the effects of the activities were calculated adjusted for the background exposure. The average personal PNC level was highest in winter and was three times higher than the mean stationary PNC level while staying indoors and two times higher while staying outdoors. Personal indoor PNC levels were significantly increased during the use of candles, cooking and the occurrence of smell of food. High stationary outdoor PNC levels and low dew point temperatures were associated with increased personal outdoor PNC levels. Times spent in public transport were associated with lower personal PNC levels than other times spent in transportation. Personal PNC levels in everyday situations exhibited a large variability because of seasonal, microenvironment-specific and activity-specific influences.


Cardiovascular autonomic neuropathy and other complications in type 1 diabetes.
This study deals with cardiovascular autonomic neuropathy (CAN) in type 1 diabetic patients and its association with other complications. We searched for CAN in 684 patients (age, 47 +/- 12 years; diabetes duration, 22 +/- 11 years) by cardiovascular responses to deep breathing and standing. Patients considered as positive had laboratory evaluation: "Ewing" tests (deep breathing, Valsalva, stand test, hand grip); heart rate variability (HRV) [low frequency (LF) and high frequency (HF) power] and spontaneous baroreflex slope (SBS). Logistic regression was used to identify the combination of patient characteristics, including other complications, most associated with CAN severity according to Ewing Score (ES 0-5). 66.2% presented no significant abnormality (ES 0-0.5), 21.5 % had mild abnormalities (ES 1-2), and 12.3% had confirmed autonomic failure (ES > 2). Decrease in LF, HF and SBS was highly correlated to CAN severity. In the stepwise regression, age, retinopathy, nephropathy, bladder dysfunction, erectile dysfunction, peripheral neuropathy and hypertension remained correlated with CAN, whereas digestive neuropathy, BMI and HbA1c were excluded. Despite a small number of events, we found a significant association between coronary disorders and CAN severity. Simple bedside tests can detect CAN. HRV and SBS provide additional elements on CAN severity. Diabetes duration did not discriminate sufficiently patients with CAN. The association with retinopathy is in favor of the role of poor glycemic control in CAN development. This study shows the interest of CAN detection and the need to look for extracardiac autonomic neuropathy and silent myocardial ischemia in patients with confirmed CAN.


Geneious! Simplified genome skimming methods for phylogenetic systematic studies: A case study in Oreocarya (Boraginaceae).
As systematists grapple with how to best harness the power of next-generation sequencing (NGS), a deluge of review papers, methods, and analytical tools make choosing the right method difficult. Oreocarya (Boraginaceae), a genus of 63 species, is a good example of a group lacking both species-level resolution and genomic resources. The use of Geneious removes bioinformatic barriers and makes NGS genome skimming accessible to even the least tech-savvy systematists. • A combination of de novo and reference-guided assemblies was used to process 100-bp single-end Illumina HiSeq 2000 reads. A subset of 25 taxa was used to test the suitability of genome skimming for future systematic studies in recalcitrant lineages like Oreocarya. • The nuclear ribosomal cistron, the plastome, and 12 mitochondrial genes were recovered from all 25 taxa. All data processing and phylogenomic analyses were performed in Geneious. We report possible future multiplexing levels and published low-copy nuclear genes represented within de novo contigs. • Genome skimming represents a much-improved primary data collection over PCR+Sanger sequencing when chloroplast DNA (cpDNA), nuclear ribosomal DNA (nrDNA), and mitochondrial DNA (mtDNA) are the target sequences. This study details methods that plant systematists can employ to study their own taxa of interest.


Ocular Signs and Ocular Comorbidities in Monoclonal Gammopathy: Analysis of 80 Subjects.
To examine the ocular signs of monoclonal gammopathy and to evaluate ocular comorbidities in subjects with monoclonal gammopathy. Patients and Methods. We analyzed patients from two large referral hematology centers in Budapest, diagnosed and/or treated with monoclonal gammopathy between 1997 and 2020. As a control group, randomly selected individuals of the same age group, without hematological disease, have been included. There were 160 eyes of 80 patients (38.75% males; age 67.61 ± 10.48 (range: 38-85) years) with monoclonal gammopathy and 86 eyes of 43 control subjects (32.56% males; age 62.44 ± 11.89 (range 37-86) years). The hematological diagnosis was MGUS in 9 (11.25%), multiple myeloma in 61 (76.25%), smoldering myeloma in 6 (7.50%), and amyloidosis or Waldenström macroglobulinemia in 2 cases (2.50%-2.50%). Before detailed ophthalmic examination with fundoscopy, 42 subjects with gammopathy (52.50%) and all controls filled the Ocular Surface Disease Index (OSDI) questionnaire. The OSDI score and best-corrected visual acuity (BCVA) were significantly worse in subjects with monoclonal gammopathy than in controls (p=0.02; p=0.0005). Among gammopathy subjects, we observed potential corneal immunoglobulin deposition in 6 eyes of 4 (3.75%) patients. Ocular surface disease (p=0.0001), posterior cortical cataract (p=0.01), and cataract (p=0.0001) were significantly more common among gammopathy subjects than in controls (χ 2 test). Ocular surface disease and cataract are more common, and BCVA is worse in patients with monoclonal gammopathy than in age-matched controls. Therefore, and due to the potential ocular signs and comorbidities of monoclonal gammopathy, we suggest a regular, yearly ophthalmic checkup of these patients to improve their quality of life.


Effect of metoclopramide on orthostatic hypotension in diabetes mellitus: failure to demonstrate the role of vasoconstrictive hormones.
Effect of metoclopramide (MCP), a dopaminergic antagonist, on orthostatic hypotension associated with diabetes mellitus, was investigated to prove its plausible role of renin-angiotensin-aldosterone and catecholamine activation by comparative study in 6 diabetics with orthostatic hypotension (OH) and 9 diabetics without OH. With MCP treatment, drop of systolic blood pressure (SBP) on standing was significantly improved. However, the postural responses of plasma renin activity (PRA), plasma aldosterone and norepinephrine were not enhanced by MCP treatment. Moreover, MCP treatment did not modify the vascular reactivity to the exogenously infused angiotensin II and norepinephrine. Thus, it would be interesting to speculate that MCP can improve SBP through its suppressive effect on some depressor substance(s) such as bradykinin which liberates exaggeratively in diabetic OH.


Stroke Risk Factors Beyond the CHA₂DS₂-VASc Score: Can We Improve Our Identification of "High Stroke Risk" Patients With Atrial Fibrillation?
The prevention of stroke and other thromboembolic events plays a crucial role in the management of patients with atrial fibrillation. Not all patients with atrial fibrillation are equal in terms of thromboembolic risk; therefore, not all will benefit from oral anticoagulation treatment. The general principle is that the expected benefit of anticoagulation in reduction of thromboembolic risk must exceed the expected harm caused by possible bleeding. Some guidelines have focused on a categorical approach to stroke prevention, with a focus on identifying patients at high risk for oral anticoagulation. Various current guidelines recommend assessment of stroke risk using the CHADS2 or CHA2DS2-VASc scores to initially detect patients at low risk who require no antithrombotic therapy. However, the scores do not incorporate all possible risk factors causing a high thromboembolic risk. Factors such as impaired renal function, obstructive sleep apnea, and echocardiographic and biochemical or coagulation parameters can also predict adverse thromboembolic events. The present review aims to describe biomarkers whether blood, urine, imaging (cardiac or cerebral), or clinical that go beyond the CHA2DS2-VASc score and potentially aid stroke risk assessment. Although useful in some cases, the presented parameters should be perhaps used to further refine initial identification of patients at low risk, after which effective stroke prevention can be offered to those with ≥1 additional stroke risk factors.