Functional identification of dendritic cells in the teleost model, rainbow trout (Oncorhynchus mykiss).
Dendritic cells are specialized antigen presenting cells that bridge innate and adaptive immunity in mammals. This link between the ancient innate immune system and the more evolutionarily recent adaptive immune system is of particular interest in fish, the oldest vertebrates to have both innate and adaptive immunity. It is unknown whether dendritic cells co-evolved with the adaptive response, or if the connection between innate and adaptive immunity relied on a fundamentally different cell type early in evolution. We approached this question using the teleost model organism, rainbow trout (Oncorhynchus mykiss), with the aim of identifying dendritic cells based on their ability to stimulate naïve T cells. Adapting mammalian protocols for the generation of dendritic cells, we established a method of culturing highly motile, non-adherent cells from trout hematopoietic tissue that had irregular membrane processes and expressed surface MHCII. When side-by-side mixed leukocyte reactions were performed, these cells stimulated greater proliferation than B cells or macrophages, demonstrating their specialized ability to present antigen and therefore their functional homology to mammalian dendritic cells. Trout dendritic cells were then further analyzed to determine if they exhibited other features of mammalian dendritic cells. Trout dendritic cells were found to have many of the hallmarks of mammalian DCs including tree-like morphology, the expression of dendritic cell markers, the ability to phagocytose small particles, activation by toll-like receptor-ligands, and the ability to migrate in vivo. As in mammals, trout dendritic cells could be isolated directly from the spleen, or larger numbers could be derived from hematopoietic tissue and peripheral blood mononuclear cells in vitro.


Development of gastric dysplasia in pernicious anaemia: a clinical and endoscopic follow up study of 80 patients.
The development of gastric dysplasia and neoplasia in patients with pernicious anaemia has been evaluated in a prospective clinical and endoscopic follow up study. After initial screening of 80 patients between 1978 and 1980, one patient underwent total gastrectomy for a gastric malignancy and 12 were kept under surveillance and underwent endoscopy at a mean interval of 14 months. In the remaining 67 patients further investigation was attempted six to seven years after the initial investigation. The continuous follow up study identified carcinoids in one patient and an adenoma with severe dysplasia in a further patient. The grade and site of mucosal dysplasia differed from one investigation to the next, but no overall progression was observed. Of the 79 patients, eight had died by the time of the reinvestigation, two of unknown cause and six of causes unrelated to pernicious anaemia. In 38 of the remaining 71 patients, clinical data only were available, with no evidence of new gastric malignancy. In 27 patients it was possible to perform an upper gastrointestinal endoscopy, when no change in the degree of dysplasia was detected. In six patients no follow up information was obtainable. In conclusion, patients with pernicious anaemia should be investigated by upper gastrointestinal endoscopy soon after diagnosis. Polyps should be removed wherever possible. In the presence of severe mucosal dysplasia or polyps that are not removed, frequent reinvestigation should be performed, provided the patient is eligible for gastric surgery. In the remaining patients, follow up endoscopies at about five year intervals would seem sufficient.


Differential effects of microRNAs miR-21, miR-99 and miR-145 on lung regeneration and inflammation during recovery from influenza pneumonia.
In a mouse model of influenza pneumonia, we previously documented that proliferating alveolar type II (AT2) cells are the major stem cells involved in early lung recovery. Profiling of microRNAs revealed significant dysregulation of specific ones, including miR-21 and miR-99a. Moreover, miR-145 is known to exhibit antagonism to miR-21. This follow-up study investigated the roles of microRNAs miR-21, miR-99a, and miR-145 in the murine pulmonary regenerative process and inflammation during influenza pneumonia. Inhibition of miR-21 resulted in severe morbidity, and in significantly decreased proliferating AT2 cells due to impaired transition from innate to adaptive immune responses. Knockdown of miR-99a culminated in moderate morbidity, with a significant increase in proliferating AT2 cells that may be linked to PTEN downregulation. In contrast, miR-145 antagonism did not impact morbidity nor the proliferating AT2 cell population, and was associated with downregulation of TNF-alpha, IL1-beta, YM1, and LY6G. Hence, a complex interplay exists between expression of specific miRNAs, lung regeneration, and inflammation during recovery from influenza pneumonia. Inhibition of miR-21 and miR-99a (but not miR-145) can lead to deleterious cellular and molecular effects on pulmonary repair and inflammatory processes during influenza pneumonia.


NLRP3 inflammasome is activated in fibromyalgia: the effect of coenzyme Q10.
Fibromyalgia (FM) is a prevalent chronic pain syndrome characterized by generalized hyperalgesia associated with a wide spectrum of symptoms such as fatigue and joint stiffness. Diagnosis of FM is difficult due to the lack of reliable diagnostic biomarkers, while treatment is largely inadequate. We have investigated the role of coenzyme Q10 (CoQ10) deficiency and mitochondrial dysfunction in inflammasome activation in blood cells from FM patients, and in vitro and in vivo CoQ10 deficiency models. Mitochondrial dysfunction was accompanied by increased protein expression of interleukin (IL)-1β, NLRP3 (NOD-like receptor family, pyrin domain containing 3) and caspase-1 activation, and an increase of serum levels of proinflammatory cytokines (IL-1β and IL-18). CoQ10 deficiency induced by p-aminobenzoate treatment in blood mononuclear cells and mice showed NLRP3 inflammasome activation with marked algesia. A placebo-controlled trial of CoQ10 in FM patients has shown a reduced NLRP3 inflammasome activation and IL-1β and IL-18 serum levels. These results show an important role for the NLRP3 inflammasome in the pathogenesis of FM, and the capacity of CoQ10 in the control of inflammasome. These findings provide new insights into the pathogenesis of FM and suggest that NLRP3 inflammasome inhibition represents a new therapeutic intervention for the disease.


[Impact of slow-release oral morphine on drug abusing habits in Austria].
A well-established possibility to treat opiate addiction is the participation in opiate maintenance treatment programmes. For this purpose the opioids methadone and buprenorphine have been evaluated and are used nowadays in many countries. However, since 1998 also the use of slow-release oral morphine (SROM) has been legally permitted in Austria. Our data show that these morphine preparations are frequently abused and are dominating the black market in the meantime. Especially the intravenous consumption of SROM goes along with highly dangerous side effects that exceed the risks of needle sharing alone. Special galenics are supposed to ensure a 24 h effect of the otherwise quickly metabolised morphine. If dissolved and injected, insoluble contents such as talcum cause microembolisms, leading to severe damages of the inner organs. Furthermore, SROM, i.e. a drug prescribed by physicians, has been proved to be the main responsible substance in most drug related deaths since its permission and has nearly replaced heroin. Forensic physicians play a major role in the profound examination of these cases, including extensive toxicological analyses and interpretation of results. For instance, a differentiation between a recent morphine and heroin consumption is certainly possible, provided appropriate methods are used. A reliable estimation of the current situation of drug abusing habits is a premise for adequate therapeutic offers and preventive measures. Thus, well-founded and comparable data have to be collected. To facilitate data report a standardized report form has been developed that includes an obligatory statement regarding morphine or heroin consumption. This should help to enlighten the ongoing discussion on the role of SRM in drug abuse cases. Our results indicate that the prescription of SROM in opiate maintenance therapy has to be handled very strictly and should be reserved for special patients only. A slackening of the Austrian law concerning SROM is therefore objected.


Crystallization and identification of an assembly defect of recombinant antenna complexes produced in transgenic tobacco plants.
A chimeric Lhcb gene encoding light-harvesting chlorophyll a/b-binding protein (LHCII) was expressed in transgenic tobacco plants. To separate native from recombinant LHCII, the protein was extended by six histidines at its C terminus. Recombinant LHCII was isolated by detergent-mediated monomerization of pure trimers followed by affinity-chromatography on Ni(2+)-NTA-agarose (NTA is nitrilotriacetic acid). Elution with imidazole yielded recombinant monomers that formed trimers readily after dilution of the detergent without further in vitro manipulations. LHCII subunits showed the typical chlorophyll a/b ratio at all steps of purification indicating no significant loss of pigments. Transgenic tobacco overexpressed amounts of recombinant protein that corresponded to about 0.7% of total LHCII. This yield suggested that expression in planta might be an alternative to the expression of eukaryotic membrane proteins in yeast. Recombinant LHCII was able to form two-dimensional crystals after addition of digalactolipids, which diffracted electrons to 3.6-A resolution. LHCII carrying a replacement of Arg-21 with Gln accumulated to only 0.004% of total thylakoid proteins. This mutant was monomeric in the photosynthetic membrane probably due to the deletion of the phosphatidylglycerol binding site and was degraded by the plastidic proteolytic system. Exchange of Asn-183 with Leu impaired LHCII biogenesis in a similar way presumably due to the lack of a chlorophyll a binding site.


Simultaneous heart and kidney transplantation in patients with end-stage heart and renal failure.
Combined simultaneous organ transplantation has become more common as selection criteria for transplantation have broadened. Broadening selection criteria is secondary to improved immunosuppression and surgical techniques. The kidney is the most common extrathoracic organ to be simultaneously transplanted with the heart. A series of 13 patients suffering from both end-stage heart and renal failure underwent 14 simultaneous heart and kidney transplantations at Temple University Hospital between 1990 and 1999. This is the largest series reported from a single center. Three patients died during the initial hospitalization for an in-hospital mortality of 21%. Of 10 patients who left the hospital, 1-year survival was 100% and 2-year survival 75%. One patient required retransplant for rejection within the first year. Overall mortality at 1 and 2 years was 25 and 41%, respectively. Four out of nine (44%) patients greater than 5 years post-transplant were alive. Of the 10 patients who left the hospital, 66% were alive at 5 years. One patient succumbed to primary nonfunction of the cardiac allograft, while the four other deaths were secondary to bacterial or fungal sepsis. The patient's racial backgrounds were equally divided between African-American and white. These results are similar to those reported in a United Network of Organ Sharing Database (UNOS) registry analysis of 84 simultaneous heart and kidney transplants that found 1- and 2-year survival to be 76 and 67%, respectively. Simultaneous heart and kidney transplantation continues to be a viable option for patients suffering from failure of these two organ systems, although the results do not match those of heart transplant alone.


[Late results of mitral recommissurotomy].
Late results of 394 operations for mitral restenosis (or 90.6% of discharged cases) were reviewed, the average duration of follow-up being 5.6 +/- 0.6 years. Late results were good in 41.9%, satisfactory in 27.2% and poor in 19.5% of the patients. Long-term mortality was 11.4%. Moderate valvular regurgitation did not significantly affect the outcome of surgery. Long-term outcomes were shown to deteriorate progressively with the increment of valvular calcinosis. The comparative actuarial analysis in long-term outcomes of mitral commissurotomies and recommissurotomies showed no significant variations. It is emphasized that mitral recommissurotomy is a promising approach to surgical treatment of mitral restenosis.


Predictive value of urinary and serum biomarkers in young children with febrile urinary tract infections.
Early predictive biomarkers for the diagnosis and management of febrile urinary tract infections (UTIs) can be valuable diagnostic tools in children. The study cohort comprised 73 pediatric patients with febrile UTIs [46 with acute pyelonephritis (APN) and 27 with lower UTIs] and 56 healthy children. Urine neutrophil gelatinase-associated lipocalin (uNGAL) and kidney injury molecule-1 (uKIM-1) levels and serum cystatin C (sCysC) levels were measured. The uNGAL/creatinine (Cr) and uKIM-1/Cr levels were higher in the UTI group than in the controls (P < 0.05). uNGAL/Cr and sCysC levels were higher in patients with APN than in those with lower UTIs (P < 0.05). uNGAL/Cr levels in both the APN and UTI groups decreased following the administration of antibiotics compared to those before treatment (P < 0.05). The uNGAL/Cr level was correlated with serum levels of white blood cells, C-reactive protein, CysC and with uKIM-1/Cr (P < 0.05). uKIM-1/Cr was also correlated with sCysC (P < 0.05). Receiver operating curve analyses showed good diagnostic profiles of uNGAL/Cr and uKIM-1/Cr for identifying UTIs [area under the curve (AUC) 0.9 and 0.66, respectively) and of uNGAL/Cr and sCysC for predicting APN (AUC 0.78 and 0.72, respectively). Our results suggest that uNGAL, uKIM-1 and sCysC levels may be useful for predicting and managing febrile UTIs in children.


Is continuous subglottic suctioning cost-effective for the prevention of ventilator-associated pneumonia?
To establish whether continuous subglottic suctioning (CSS) could be cost-effective. Cost-benefit analysis, based on a hypothetical replacement of conventional ventilation (CV) with CSS. A surgical intensive care unit (SICU) of a tertiary care university hospital in France. All consecutive patients receiving ventilation in the SICU in 2006. Efficacy data for CSS were obtained from the literature and applied to the SICU of our hospital. Costs for CV and CSS were provided by the hospital pharmacy; costs for ventilator-associated pneumonia (VAP) were obtained from the literature. The cost per averted VAP episode was calculated, and a sensitivity analysis was performed on VAP incidence and on the number of tubes required for each patient. At our SICU in 2006, 416 patients received mechanical ventilation for 3,487 ventilation-days, and 32 VAP episodes were observed (7.9 episodes per 100 ventilated patients; incidence density, 9.2 episodes per 10,000 ventilation-days). Based on the hypothesis of a 29% reduction in the risk of VAP with CSS than CV, 9 VAP episodes could have been averted. The additional cost of CSS for 2006 was estimated to be €10,585.34. The cost per averted VAP episode was €1,176.15. Assuming a VAP cost of €4,387, a total of 3 averted VAP episodes would neutralize the additional cost. For a low VAP incidence of 6.6%, the cost per averted VAP would be €1,323. If each patient required 2 tubes during ventilation, the cost would be €1,383.69 per averted VAP episode. Replacement of CV with CSS was cost-effective even when assuming the most pessimistic scenario of VAP incidence and costs.


Organic electronic ratchets doing work.
The possibility to extract work from periodic, undirected forces has intrigued scientists for over a century—in particular, the rectification of undirected motion of particles by ratchet potentials, which are periodic but asymmetric functions. Introduced by Smoluchowski and Feynman to study the (dis)ability to generate motion from an equilibrium situation, ratchets operate out of equilibrium, where the second law of thermodynamics no longer applies. Although ratchet systems have been both identified in nature and used in the laboratory for the directed motion of microscopic objects, electronic ratchets have been of limited use, as they typically operate at cryogenic temperatures and generate subnanoampere currents and submillivolt voltages. Here, we present organic electronic ratchets that operate up to radio frequencies at room temperature and generate currents and voltages that are orders of magnitude larger. This enables their use as a d.c. power source. We integrated the ratchets into logic circuits, in which they act as the d.c. equivalent of the a.c. transformer, and generate enough power to drive the circuitry. Our findings show that electronic ratchets may be of actual use.


Fine mapping and characterization of candidate lung tumor resistance genes for the Par2 locus on mouse chromosome 18.
In a number of recent studies, a lung tumor resistance locus designated either Par2 or Pas7 was mapped to distal chromosome 18 in crosses between susceptible A/J and more resistant BALB/c mice. This locus is important in that it accounts for as much as 60% of the difference in lung tumor susceptibility between the A/J and BALB/c mice, both of which contain the susceptible allele of Kras2, a marker and strong candidate for the major lung tumor susceptibility gene on mouse chromosome 6. We have now fine-mapped the Par2 locus by using congenic mice that were constructed by placing part of chromosome 18 from the susceptible A/J onto the genetic background of lung tumor-resistant BALB/c mice. After 7 generations of backcrossing, N7 mice that carried 28 cM of the A/J quantitative trait locus (QTL) region were crossed to the BALB/c to generate the N8 generation. Congenic strains (N8) that contain various QTL regions were generated. N9 mice, generated from N8 males x 3 BALB/c females, were genotyped in the region of the Par2 locus and treated with an initiating dose of urethane and allowed to form lung tumors over 6 months. The mice were killed and the lung tumors counted. With this cross the Par2 locus was narrowed to a 6-cM region. Potential candidate genes in this region include Smad4, Smad2, and Dcc. Previously, we excluded Smad4 and Smad2 as candidates for Par2 based on the lack of functional polymorphism(s) and differential expression in lungs from A/J and BALB/c mice. In this study, no polymorphism of the coding sequence of Dcc was observed between A/J and BALB/c mice. Further fine mapping and positional cloning are required for the identification of the Par2 gene.


Targeting of Wolbachia endobacteria in Litomosoides sigmodontis: comparison of tetracyclines with chloramphenicol, macrolides and ciprofloxacin.
Endobacteria of the genus Wolbachia in filarial nematodes are related to Rickettsiaceae and can be depleted by tetracycline antibiotics. This depletion blocks female worm development as well as early embryogenesis, in contrast to the currently used microfilaricidal ivermectin which blocks only the last stage of embryogenesis. Since targeting Wolbachia is becoming an area of research for the treatment of human filariases, it was investigated if antibiotics other than tetracyclines are able to deplete Wolbachia from filariae. BALB/c mice infected with the rodent filaria Litomosoides sigmodontis were treated with erythromycin, chloramphenicol or ciprofloxacin. All drugs were well resorbed and resulted in serum levels clearly above breakpoint levels for bacteria susceptible to the respective antibiotic. However, contrary to tetracycline, none of these antibiotics depleted Wolbachia or altered worm development and fertility, as evidenced by immunohistology, immunoelectron microscopy and semiquantitative PCR.


Adopting a surgical safety checklist could save money and improve the quality of care in U.S. hospitals.
Use of the World Health Organization's Surgical Safety Checklist has been associated with a significant reduction in major postoperative complications after inpatient surgery. We hypothesized that implementing the checklist in the United States would generate cost savings for hospitals. We performed a decision analysis comparing implementation of the checklist to existing practice in U.S. hospitals. In a hospital with a baseline major complication rate after surgery of at least 3 percent, the checklist generates cost savings once it prevents at least five major complications. Using the checklist would both save money and improve the quality of care in hospitals throughout the United States.


Quality of Pregnancy Dating and Obstetric Interventions During Labor: Retrospective Database Analysis.
The correct dating of pregnancy is critical to support timely decisions and provide obstetric care during birth. The early obstetric ultrasound assessment before 14 weeks is considered the best reference to assist in determining gestational age (GA), with an accuracy of ±5 to 7 days. However, this information is limited in many settings worldwide. The aim of this study is to analyze the association between the obstetric interventions during childbirth and the quality of GA determination, according to the first antenatal ultrasound assessment, which assisted the calculation. This is a hospital-based cohort study using medical record data of 2113 births at a perinatal referral center. The database was separated into groups and subgroups of analyses based on the reference used by obstetricians to obtain GA at birth. Maternal and neonatal characteristics, mode of delivery, oxytocin augmentation, and forceps delivery were compared between groups of pregnancies with GA determination at different reference points: obstetric ultrasound assessment 14 weeks, 20 weeks, and ≥20 weeks or without antenatal ultrasound (suboptimal dating). Ultrasound-based GA information was associated with outcomes between the interest groups using chi-square tests, odds ratios (OR) with 95% CI, or the Mann-Whitney statistical analysis. The chance of nonspontaneous delivery was higher in pregnancies with 14 weeks ultrasound-based GA (OR 1.64, 95% CI 1.35-1.98) and 20 weeks ultrasound-based GA (OR 1.58, 95% CI 1.31-1.90) when compared to the pregnancies with ≥20 weeks ultrasound-based GA or without any antenatal ultrasound. The use of oxytocin for labor augmentation was higher for 14 weeks and 20 weeks ultrasound-based GA, OR 1.41 (95% CI 1.09-1.82) and OR 1.34 (95% CI 1.04-1.72), respectively, when compared to those suboptimally dated. Moreover, maternal blood transfusion after birth was more frequent in births with suboptimal ultrasound-based GA determination (20/657, 3.04%) than in the other groups (14 weeks ultrasound-based GA: 17/1163, 1.46%, P=.02; 20 weeks ultrasound-based GA: 25/1456, 1.71%, P=.048). Cesarean section rates between the suboptimal dating group (244/657, 37.13%) and the other groups (14 weeks: 475/1163, 40.84%, P=.12; 20 weeks: 584/1456, 40.10%, P=.20) were similar. In addition, forceps delivery rates between the suboptimal dating group (17/657, 2.6%) and the other groups (14 weeks: 42/1163, 3.61%, P=.24; 20 weeks: 46/1456, 3.16%, P=.47) were similar. Neonatal intensive care unit admission was more frequent in newborns with suboptimal dating (103/570, 18.07%) when compared with the other groups (14 weeks: 133/1004, 13.25%, P=.01; 20 weeks: 168/1263, 13.30%, P=.01), excluding stillbirths and major fetal malformations. The present analysis highlighted relevant points of health care to improve obstetric assistance, confirming the importance of early access to technologies for pregnancy dating as an essential component of quality antenatal care.


[Behavior of calcium, phosphorus and magnesium in rats subjected to malnutrition in different stages of the gestational period].
An attempt was made to point out the alterations in serum calcium, phosphorus and magnesium, during gestation in female rats fed ad libitum normal diets (protein content, 21%), a protein-calorie deficient diet (protein content, 1%) in a quantity equal to half of that given to normal rats. As the data revealed, in the control group of pregnant rats a significant increase in calcium on the 17th and 19th days was observed, as well as a decrease at the end of gestation. The values remained at higher levels in this group than those detected in the control non-pregnant rats. The above-mentioned increase was not observed in the undernourished pregnant rats, which showed an evident decrease. The behavior of phosphorus in the control pregnant rats was similar to calcium behavior, but this "ion" had not suffered the influence of malnutrition when isolatedly imposed. With regard to magnesium, we noticed that throughout the experiment serum concentration was not influenced either by malnutrition or by gestation. Malnutrition itself was the greatest responsible factor as judged by the differences observed among the groups.


The impact of the Hazelwood coal mine fire smoke exposure on asthma.
In 2014, a fire at an open cut coal mine in South-eastern Australia burned for about 6 weeks. Residents of the adjacent town were exposed to high levels of fine particulate matter (PM2.5) during this period. Three and a half years after the event, this study aimed to investigate potential long-term impacts of short-term exposure to coal mine fire smoke on asthma. A cross-sectional analysis was undertaken in a group of exposed participants with asthma from Morwell (n = 165) and a group of unexposed participants with asthma from the control town of Sale (n = 64). Exposure was determined by modeled PM2.5 data for the mine fire period. Respiratory symptoms were assessed with a validated respiratory health questionnaire and symptom severity score. Asthma control was assessed with a validated questionnaire. Lung function testing included spirometry, bronchodilator response, and fraction of exhaled nitric oxide. There was no evidence that exposed participants had more severe asthma symptoms, worse lung function, or more eosinophilic airway inflammation than unexposed participants. However, there was some evidence that Morwell participants had more uncontrolled than well-controlled asthma, compared to the participants from Sale (adjusted relative risk ratio 2.71 95% CI: 1.02, 7.21, p = .046). Three and a half years after exposure, coal mine fire smoke did not appear to be associated with more severe asthma symptoms or worse lung function but might be associated with poorer asthma control.


Flexible transparent displays based on core/shell upconversion nanophosphor-incorporated polymer waveguides.
Core/shell (C/S)-structured upconversion nanophosphor (UCNP)-incorporated polymer waveguide-based flexible transparent displays are demonstrated. Bright green- and blue-emitting Li(Gd,Y)F4:Yb,Er and Li(Gd,Y)F4:Yb,Tm UCNPs are synthesized via solution chemical route. Their upconversion luminescence (UCL) intensities are enhanced by the formation of C/S structure with LiYF4 shell. The Li(Gd,Y)F4:Yb,Er/LiYF4 and Li(Gd,Y)F4:Yb,Tm/LiYF4 C/S UCNPs exhibit 3.3 and 2.0 times higher UCL intensities than core counterparts, respectively. In addition, NaGdF4:Yb,Tm/NaGdF4:Eu C/S UCNPs are synthesized and they show red emission via energy transfer and migration of Yb3+ → Tm3+ → Gd3+ → Eu3+. The C/S UCNPs are incorporated into bisphenol A ethoxylate diacrylate which is used as a core material of polymer waveguides. The fabricated stripe-type polymer waveguides are highly flexible and transparent (transmittance > 90% in spectral range of 443-900 nm). The polymer waveguides exhibit bright blue, green, and red luminescence, depending on the incorporated UCNPs into the polymer core, under coupling with a near infrared (NIR) laser. Moreover, patterned polymer waveguide-based display devices are fabricated by reactive ion etching process and they realize bright blue-, green-, and red-colored characters under coupling with an NIR laser.


Toward a Standard Protocol for Micelle Simulation.
In this paper, we present protocols for simulating micelles using dissipative particle dynamics (and in principle molecular dynamics) that we expect to be appropriate for computing micelle properties for a wide range of surfactant molecules. The protocols address challenges in equilibrating and sampling, specifically when kinetics can be very different with changes in surfactant concentration, and with minor changes in molecular size and structure, even using the same force field parameters. We demonstrate that detection of equilibrium can be automated and is robust, for the molecules in this study and others we have considered. In order to quantify the degree of sampling obtained during simulations, metrics to assess the degree of molecular exchange among micellar material are presented, and the use of correlation times are prescribed to assess sampling and for statistical uncertainty estimates on the relevant simulation observables. We show that the computational challenges facing the measurement of the critical micelle concentration (CMC) are somewhat different for high and low CMC materials. While a specific choice is not recommended here, we demonstrate that various methods give values that are consistent in terms of trends, even if not numerically equivalent.


Rotatable reagent cartridge for high-performance microvalve system on a centrifugal microfluidic device.
Recently, microfluidic lab-on-a-CD (LabCD) has attracted attentions of researchers for its potential for pumpless, compact, and chip-inclusive on-site bioassay. To control the fluids in the LabCD, microvalves such as capillary, hydrophobic, siphon, and sacrificial valves have been employed. However, no microvalve can regulate more than one channel. In a complicated bioassay with many sequential mixing, washing, and wasting steps, thus, an intricate fluidic network with many microchannels, microvalves, and reservoirs is required, which increases assay costs in terms of both system development and chip preparation. To address this issue, we developed a rotatable reagent cartridge (RRC), which was a column-shaped tank and has several rooms to store different reagents. By embedding and rotating the RRC in the LabCD with a simple mechanical force, only the reagent in the room connected to the following channel was injected. By regulating the angle of the RRC to the LabCD, conservation and ejection of each reagent could be switched. Our developed RRC had no air vent hole, which was achieved by the gas-permeable gap between the bottle and cap parts of the RRC. The RRC could inject 230 nL-10 μL of reagents with good recoveries more than 96%. Finally, an enzymatic assay of L-lactate was demonstrated, where the number of valves and reservoirs were well minimized, significantly simplifying the fluidic system and increasing the channel integratability. Well quantitative analyses of 0-100 μM L-lactate could easily be carried out with R(2) > 0.999, indicating the practical utility of the RRC for microfluidic bioanalysis.


Four Aromatic Intradiol Ring Cleavage Dioxygenases from Aspergillus niger.
Ring cleavage dioxygenases catalyze the critical ring-opening step in the catabolism of aromatic compounds. The archetypal filamentous fungus Aspergillus niger previously has been reported to be able to utilize a range of monocyclic aromatic compounds as sole sources of carbon and energy. The genome of A. niger has been sequenced, and deduced amino acid sequences from a large number of gene models show various levels of similarity to bacterial intradiol ring cleavage dioxygenases, but no corresponding enzyme has been purified and characterized. Here, the cloning, heterologous expression, purification, and biochemical characterization of four nonheme iron(III)-containing intradiol dioxygenases (NRRL3_02644, NRRL3_04787, NRRL3_05330, and NRRL3_01405) from A. niger are reported. Purified enzymes were tested for their ability to cleave model catecholate substrates, and their apparent kinetic parameters were determined. Comparisons of kcat/Km values show that NRRL3_02644 and NRRL3_05330 are specific for hydroxyquinol (1,2,4-trihydroxybenzene), and phylogenetic analysis shows that these two enzymes are related to bacterial hydroxyquinol 1,2-dioxygenases. A high-activity catechol 1,2-dioxygenase (NRRL3_04787), which is phylogenetically related to other characterized and putative fungal catechol 1,2-dioxygenases, was also identified. The fourth enzyme (NRRL3_01405) appears to be a novel homodimeric Fe(III)-containing protocatechuate 3,4-dioxygenase that is phylogenetically distantly related to heterodimeric bacterial protocatechuate 3,4-dioxygenases. These investigations provide experimental evidence for the molecular function of these proteins and open the way to further investigations of the physiological roles for these enzymes in fungal metabolism of aromatic compounds.IMPORTANCE Aromatic ring opening using molecular oxygen is one of the critical steps in the degradation of aromatic compounds by microorganisms. While enzymes catalyzing this step have been well-studied in bacteria, their counterparts from fungi are poorly characterized despite the abundance of genes annotated as ring cleavage dioxygenases in fungal genomes. Aspergillus niger degrades a variety of aromatic compounds, and its genome harbors 5 genes encoding putative intracellular intradiol dioxygenases. The ability to predict the substrate specificities of the encoded enzymes from sequence data are limited. Here, we report the characterization of four purified intradiol ring cleavage dioxygenases from A. niger, revealing two hydroxyquinol-specific dioxygenases, a catechol dioxygenase, and a unique homodimeric protocatechuate dioxygenase. Their characteristics, as well as their phylogenetic relationships to predicted ring cleavage dioxygenases from other fungal species, provide insights into their molecular functions in aromatic compound metabolism by this fungus and other fungi.


Characterization of a tobacco Bright Yellow 2 cell line expressing the tetracycline repressor at a high level for strict regulation of transgene expression.
Manipulating the expression of a transgene in transient and stable transformed cells is a requirement for many functional analyses. We have investigated the use of the tetracycline-dependent gene expression system developed by Gatz et al. (1992) in tobacco (Nicotiana tabacum L. cv Bright Yellow 2 [BY2]) cells, the most widely used plant cell culture. We have selected a BY2 cell line, named BY2-tetracycline repressor (tetR) 17, which expresses the tetR at a high level, and have evaluated the capacity of this cell line to suppress the expression of a green fluorescent protein reporter gene under the control of the "Triple-Op" promoter in the absence of tetracycline in a large number of independent transformants. The ability to induce the expression of green fluorescent protein after treatment by anhydrotetracycline in the same transformants was also analyzed. BY2-tetR17 cells were demonstrated to be excellent recipient cells for recovery of clonal cell lines with a highly controlled regulation of the introduced transgene.


A rare cause of bile duct obstruction in adolescence: Neuroendocrine tumor.
Extrahepatic bile duct is one of the rare places for neuroendocrine tumors which comprise 0.2-2% off all neuroendocrine tumors of gastrointestinal tract. The aim of this paper is to report a case of a 16-year-old boy with a neuroendocrine tumor of extra hepatic bile duct. Laboratory and imaging findings is supported obstructive jaundice. After a pre-operative detailed evaluation, the common bile and common hepatic duct with gallbladder were resected and Roux-en-Y hepaticojejunostomy was performed. After a 36 months follow up, no locoregional recurrence or metastatic disease was observed. Pre-operative diagnosis of neuroendocrine tumors are extremely difficult and often confused with cholangiocarcinoma. Treatment modality of this rare entity depends on the location of the tumor and aggressive surgery is still the best chocie of treatment. It should be kept in mind that disease-free survival for patients with neuroendocrine tumor depends on success of surgery.


Differential control of peripheral circadian rhythms by suprachiasmatic-dependent neural signals.
Although dependent on the integrity of a central pacemaker in the suprachiasmatic nucleus of the hypothalamus (SCN), endogenous daily (circadian) rhythms are expressed in a wide variety of peripheral organs. The pathways by which the pacemaker controls the periphery are unclear. Here, we used parabiosis between intact and SCN-lesioned mice to show that nonneural (behavioral or bloodborne) signals are adequate to maintain circadian rhythms of clock gene expression in liver and kidney, but not in heart, spleen, or skeletal muscle. These results indicate that the SCN regulates expression of circadian oscillations in different peripheral organs by diverse pathways.


Positional effects on exophthalmometer readings in Graves' eye disease.
Forty-eight patients with Graves' eye disease, half of whom had extraocular muscle restriction, and 45 normal adult subjects underwent exophthalmometry while in the upright and the supine positions to determine whether the difference in measurement due to position would serve to differentiate the groups. A previous report claimed that the upright reading would be greater than the supine reading for normal subjects and patients with Graves' eye disease without clinical extraocular muscle involvement, whereas those with extraocular muscle restriction would show little change in their exophthalmometer readings. Our study showed an increase in exophthalmometer readings in going from the upright to the supine position for normal subjects and both groups of patients with Graves' eye disease. Furthermore, this test did not serve to differentiate these three groups.


Mental health inequalities among gay and bisexual men in England, Scotland and Wales: a large community-based cross-sectional survey.
Sexual minorities suffer worse mental health than the sexual majority but little is known about differences in mental health within sexual minorities. We aimed to describe inequality in mental health indicators among gay and bisexual men. Using multi-channel community-based opportunistic sampling we recruited 5799 eligible men aged 16 years and over, living in England, Scotland and Wales and who were sexually attracted to other men, to a self-completion Internet health survey. Mental health indicators (depression (PHQ-9), anxiety (GAD-7), suicide attempt and self-harm) were examined for independent associations across common axes of inequality (age, ethnicity, migrancy, education, income, cohabitation and living in London). Mental ill-health was common: 21.3% were depressed and 17.1% anxious, while 3.0% had experienced attempted suicide and 6.5% had self-harmed within the last 12 months. All four indicators were associated with younger age, lower education and lower income. Depression was also associated with being a member of visible ethnic minorities and sexual attraction to women as well as men. Cohabiting with a male partner and living in London were protective of mental health. Community interventions to increase mental health among gay and bisexual men should be designed to disproportionately benefit younger men and those living on lower incomes.


Placental Membrane Provides Improved Healing Efficacy and Lower Cost Versus a Tissue-Engineered Human Skin in the Treatment of Diabetic Foot Ulcerations.
Aseptically processed dehydrated human amnion and chorion allograft (dHACA) (AmnioBand) has shown great promise in the treatment of recalcitrant diabetic foot ulcers (DFUs) when compared with standard wound care but has not yet been compared to any other tissue forms used in treating DFUs. The hypothesis was to conduct a randomized controlled trial in which dHACA was compared to one of the earliest and most commonly accepted tissue-engineered skin substitutes (TESS) (Apligraf) in the treatment of nonhealing DFUs over a period of 12 weeks to assess the superiority of healing. Following a 2-week screening period during which subjects with DFUs were treated with collagen alginate dressing, 60 subjects were randomized at 5 sites to receive either dHACA or TESS applied weekly, with weekly follow-up for up to 12 weeks. The mean time to heal within 6-week time period for the dHACA group was 24 days (95% CI, 18.9-29.2) versus 39 days (95% CI, 36.4-41.9) for the TESS group; the mean time to heal at 12 weeks was 32 days (95% CI, 22.3-41.0) for dHACA-treated wounds versus 63 days (95% CI, 54.1-72.6) for TESS-treated wounds. The proportion of wounds healed at study completion (12 weeks) was 90% (27/30) for the dHACA group versus 40% (12/30) for the TESS group. The mean product cost for the dHACA group was significantly lower than that for the TESS group [dHACA: $2,200 (median: $1,300); TESS: $7,900 (median: $6,500)]. The mean wastage (%) at 12 weeks was also significantly lower for the dHACA group than that for the TESS group (36% vs 95%). It was concluded that aseptically processed dHACA heals diabetic foot wounds more reliably, statistically significantly faster than and at significantly lower cost than TESS.


[Coronary surgery--arterial grafts instead of venous grafts?].
Recurrence of symptoms and the need for repeat interventions remains a clinical challenge following coronary artery bypass surgery, despite excellent early results. Saphenous vein graft failure has been identified as a main contributing factor to unsatisfactory long-term results. The use of multiple arterial grafts instead of venous grafts appears to be a promising treatment modality. This article describes our own experience with arterial revascularisation and gives a critical review of the literature. Extended arterial revascularisation was performed in 30 patients at our institution between 1998 and 2001. Clinical follow-up was performed in all patients; re-angiography was done in six patients. 23 patients were operated on with bilateral mammary arteries; 11 patients received radial artery grafts. The median follow up was 12 months. All patients are alive, none suffered a new myocardial infarction. Our results are in accordance with the published literature. Arterial grafting is a valuable tool in the armamentarium of modern coronary artery bypass surgery. Large randomised trials are required to clarify the role of extended arterial grafting in routine coronary bypass surgery.


Structural and mechanical adaptations of right ventricle free wall myocardium to pressure overload.
Right ventricular (RV) failure in response to pulmonary hypertension (PH) is a severe disease that remains poorly understood. PH-induced pressure overload leads to changes in the RV free wall (RVFW) that eventually results in RV failure. While the development of computational models can benefit our understanding of the onset and progression of PH-induced pressure overload, detailed knowledge of the underlying structural and biomechanical events remains limited. The goal of the present study was to elucidate the structural and biomechanical adaptations of RV myocardium subjected to sustained pressure overload in a rat model. Hemodynamically confirmed severe chronic RV pressure overload was induced in Sprague-Dawley rats via pulmonary artery banding. Extensive tissue-level biaxial mechanical and histomorphological analyses were conducted to assess the remodeling response in the RV free wall. Simultaneous myofiber hypertrophy and longitudinal re-orientation of myo- and collagen fibers were observed, with both fiber types becoming more highly aligned. Transmural myo- and collagen fiber orientations were co-aligned in both the normal and diseased state. The overall tissue stiffness increased, with larger increases in longitudinal vs. circumferential stiffness. The latter was attributed to longitudinal fiber re-orientation, which increased the degree of anisotropy. Increased mechanical coupling between the two axes was attributed to the increased fiber alignment. Interestingly, estimated myofiber stiffness increased while the collagen fiber stiffness remained unchanged. The increased myofiber stiffness was consistent with clinical results showing titin-associated increased sarcomeric stiffening observed in PH patients. These results further our understanding of the underlying adaptive and maladaptive remodeling mechanisms and may lead to improved techniques for prognosis, diagnosis, and treatment for PH.


Epigenetics and Mitochondrial Stability in the Metabolic Memory Phenomenon Associated with Continued Progression of Diabetic Retinopathy.
Retinopathy continues to progress even when diabetic patients try to control their blood sugar, but the molecular mechanism of this 'metabolic memory' phenomenon remains elusive. Retinal mitochondria remain damaged and vicious cycle of free radicals continues to self-propagate. DNA methylation suppresses gene expression, and diabetes activates DNA methylation machinery. Our aim was to investigate the role of DNA methylation in continued compromised mitochondrial dynamics and genomic stability in diabetic retinopathy. Using retinal endothelial cells, incubated in 20 mM glucose for four days, followed by 5 mM glucose for four days, and retinal microvessels from streptozotocin-induced diabetic rats in poor glycemia for four months, followed by normal glycemia for four additional months, DNA methylation of mitochondrial fusion and mismatch repair proteins, Mfn2 and Mlh1 respectively, was determined. Retinopathy was detected in trypsin-digested microvasculature. Re-institution of good glycemia had no beneficial effect on hypermethylation of Mfn2 and Mlh1 and retinal function (electroretinogram), and the retinopathy continued to progress. However, intervention of good glycemia directly with DNA methylation inhibitors (Azacytidine or Dnmt1-siRNA), prevented Mfn2 and Mlh1 hypermethylation, and ameliorated retinal dysfunction and diabetic retinopathy. Thus, direct regulation of DNA methylation can prevent/reverse diabetic retinopathy by maintaining mitochondrial dynamics and DNA stability, and prevent retinal functional damage.


Lycium barbarum polysaccharides restore adverse structural remodelling and cardiac contractile dysfunction induced by overexpression of microRNA-1.
MicroRNA-1 (miR-1) stands out as the most prominent microRNA (miRNA) in regulating cardiac function and has been perceived as a new potential therapeutic target. Lycium barbarum polysaccharides (LBPs) are major active constituents of the traditional Chinese medicine based on L. barbarum. The purpose of this study was to exploit the cardioprotective effect and molecular mechanism of LBPs underlying heart failure. We found that LBPs significantly reduced the expression of myocardial miR-1. LBPs improved the abnormal ECG and indexes of cardiac functions in P-V loop detection in transgenic (Tg) mice with miR-1 overexpression. LBPs recovered morphological changes in sarcomeric assembly, intercalated disc and gap junction. LBPs reversed the reductions of CaM and cMLCK, the proteins targeted by miR-1. Similar trends were also obtained in their downstream effectors including the phosphorylation of MLC2v and both total level and phosphorylation of CaMKII and cMyBP-C. Collectively, LBPs restored adverse structural remodelling and improved cardiac contractile dysfunction induced by overexpression of miR-1. One of the plausible mechanisms was that LBPs down-regulated miR-1 expression and consequently reversed miR-1-induced repression of target proteins relevant to myocardial contractibility. LBPs could serve as a new, at least a very useful adjunctive, candidate for prevention and therapy of heart failure.


[Detection of anti-Azotobacter antibodies in sera from tumor patients].
Employing the agar-gel double immunodiffusion technique, the authors describe the presence of antibodies to a protein fraction existing in the supernatant of Azotobacter chroococcum cultures in a high percentage of blood sera from cancer patients. The small number of false positive results makes them think of the convenience of increasing the sensitivity of the procedure to valorate its usefulness for the early diagnosis of malignant tumors.


Calcium supplementation does not alter lipid oxidation or lipolysis in overweight/obese women.
Based on cell culture and studies in mice, increased dietary calcium appears to stimulate lipolysis and could possibly reduce body adiposity through hormonal influences on adipocyte calcium uptake. In this study, we investigated the effects of 1,500 mg supplemental calcium daily for 3 months on hormones regulating calcium and energy metabolism and rates of lipid oxidation and lipolysis in overweight women. Fifteen overweight (BMI > 25 kg/m(2)) premenopausal women were supplemented with 1,500 mg of calcium, as CaCO(3), per day for 3 months while maintaining their usual diets and activity levels. Baseline and endpoint measurements were obtained after the subjects consumed a standardized 25% fat diet for 4 days. Lipid oxidation was measured by indirect calorimetry, lipolysis by infusion of deuterated glycerol, and body fat by dual-energy X-ray absorptiometry. Urinary calcium, circulating levels of hormones involved in energy and lipid metabolism (insulin, leptin, and adiponectin) or calcium metabolism (25(OH)D, 1,25(OH)(2)D), and parathyroid hormone (PTH)) were also measured. Urinary levels of calcium (P = 0.005) increased and 1,25(OH)(2)D declined (P = 0.03). However other parameters, including body weight, body fat, PTH, insulin, leptin, adiponectin, 25(OH)D, as well as rates of lipid oxidation and lipolysis were not altered by calcium supplementation. Calcium supplementation for 3 months increased urinary calcium excretion, decreased circulating levels of 1,25(OH)(2)-D, but had no effect on rates of lipid oxidation or lipolysis, in these overweight women.


Impact of a history of diabetes mellitus on quality of life after coronary artery bypass grafting.
To describe the improvement in various aspects of quality of life (QoL) after coronary artery bypass grafting (CABG), in relation to a previous history of diabetes mellitus. All patients from western Sweden who underwent CABG between 1988 and 1991 without simultaneous valve surgery. Patients were approached with three questionnaires: The Physical Activity Score, the Nottingham Health Profile and the Psychological General Well-being Index prior to surgery and 3 months, 1 and 2 years thereafter. All three questionnaires already showed a significant improvement in QoL after 3 months, remaining at a similar level 1 and 2 years after the operation. In terms of Physical Activity Score improvement was of similar magnitude in diabetic and non-diabetic patients. In terms of the Psychological General Well-Being Index significant and similar improvements were found in diabetic and non-diabetic patients at each evaluation. In terms of the Nottingham Health Profile there was a significant improvement both in diabetic and non-diabetic patients 3 months, 1 and 2 years after the operation. However, improvement was more marked in diabetic than in non-diabetic patients at each evaluation. For 3 months, 1 and 2 years after CABG various aspects of QoL as estimated with three different instruments, improved significantly both in diabetic and in non-diabetic patients compared with the situation prior to the operation. However, the three instruments differed somewhat. Thus, whereas in the Physical Activity Score, diabetic patients tended to improve less markedly than non-diabetic patients, the opposite was found in the Nottingham Health Profile.


Udder firmness as a possible indicator for clinical mastitis.
Swelling of the mammary gland is an important sign to detect clinical mastitis (CM) in dairy cows. The overall objective of this study was to evaluate if udder firmness can be used as a cow-side indicator for mastitis and to evaluate how CM affects firmness within 14 d after diagnosis. A dynamometer was used to objectively determine udder firmness before and after milking in 45 cows with CM and 95 healthy cows. Udder firmness of both hind quarters was measured daily on 3 locations (upper, middle, lower measuring point) from the day of mastitis diagnosis until d 7 and again on d 14. Firmness of the middle measuring point was highest before and after milking in all cows. Udder firmness before milking was similar in quarters without and with CM. Subsequently, we concentrated on firmness measured on the middle point after milking. After milking, quarters with CM were firmer than healthy quarters. An increase of firmness of a quarter with mastitis did not affect firmness of the healthy neighboring quarter, nor did firmness of all healthy quarters differ. One firmness value per cow [i.e., Δ firmness (difference in udder firmness between both hind quarters)] was used for all further calculations. After fitting a generalized mixed model, CM affected Δ firmness in all cases. In multiparous cows, Δ firmness was also affected by continuous milk yield per day and DIM. Firmness thresholds for detection of CM were calculated using receiver operation characteristic curves. The threshold for detection of CM using Δ firmness was 0.282 kg (area under the curve: 0.722, sensitivity: 64.3%, specificity: 89.7%) and 0.425 kg (area under the curve: 0.817, sensitivity: 62.5%, specificity: 96.7%) in primiparous cows and multiparous cows, respectively. Linear mixed-model ANOVA were used to evaluate how CM affects udder firmness within 14 d after diagnosis. Cows with CM had a higher Δ firmness compared with cows without CM throughout the 14 d after the mastitis diagnoses. Parity had an effect on Δ firmness. Depending on systemic signs of sickness, mastitic cows were divided into cows having mild to moderate (n = 21) or severe mastitis (n = 24). Cows with severe mastitis suffered from a firmer udder on all measuring days. Bacteriological cure was defined based on 2 milk samples taken at 7 and 14 d after enrollment. An effect of parity and bacteriological cure on Δ firmness of cows with CM did not exist within the 14 d. Cows not clinically cured showed an increased Δ firmness of 0.560 kg compared with cured cows. In conclusion, udder firmness can be a useful indicator for CM. Further research is warranted to evaluate if udder firmness can be used as a predictor for the prognosis of a CM or the cure of inflammation.


Global epidemiology of early-onset liver cancer attributable to specific aetiologies and risk factors from 2010 to 2019.
Considering its emergence as a public health concern worldwide, with potential spatial-temporal heterogeneities, we aimed to determine the global burden of early-onset liver cancer attributable to aetiologies and concomitant risk factors. We used data from the Global Burden of Diseases Study 2019 to determine age-standardised disability-adjusted life-year (DALY) rates for early-onset liver cancer by aetiologies and the population DALYs attributable to concomitant risk factors between 2010 and 2019. We also calculated estimated annual percentage changes (EAPCs) to measure temporal trends. There were 2.9 million DALYs related to early-onset liver cancer globally in 2019. East Asia contributed over half of DALYs, which increased annually by 1.23% (95% confidence interval (CI) = 0.71, 1.76) between 2010 and 2019. Non-alcoholic steatohepatitis was the only growing aetiology. The proportion of DALYs attributed to metabolic risks increased by 22.50% (95% CI = 14.33, 38.13), while behavioral risks remained stable. Obesity surpassed smoking as the most prevalent nondeterministic aetiological risk factor from 2010 to 2019, while the population DALY attributable to hepatitis B combined with obesity increased by 29.93% (95% CI = 8.49, 60.77) in the same period, making it the principal joint contributor. Early-onset liver cancer poses considerable disability and continues to increase in many regions, especially in East Asia. Metabolic risk factors, particularly when hepatitis B and obesity coexist, are the fastest-growing contributors to this type of cancer. More targeted interventions are imperative to curb the growing burden of early-onset liver cancer due to metabolic risks.


Transnational Tobacco Companies and New Nicotine Delivery Systems.
While the public health community has focused on the harm-reduction potential of new nicotine delivery systems (NNDSs) and, conversely, their potential for impeding overall efforts to prevent and reduce tobacco use, limited analysis has been conducted on the role of leading transnational tobacco companies (TTCs) in this rapidly growing market. Following aborted efforts during the 1980s and 1990s to develop reduced-risk products, TTCs have heavily invested in selected NNDS products since 2010 via acquisitions and internal research and development. This article catalogs and analyzes the patterns of investment in NNDSs by leading TTCs over time, and identifies differences in the companies' approaches to NNDS product acquisition and development in specific markets globally. This analysis raises important questions regarding the companies' intent, which appears to be to sustain, rather than replace, cigarette sales, and to increase their influence and credibility with respect to NNDS policy and regulation. We identify the need for greater public health vigilance and research to understand and respond to the increasingly significant role of NNDSs in TTCs' global business strategies, to ensure that NNDSs advance, rather than hinder, tobacco control efforts.


A rare case of rynopharyngeal melanoma.
Primary mucosal melanomas (MM) of the head and neck region constitute 0.5-2% of all malignant melanomas. The rynopharynx is a region that is less often involved by malignant melanomas. Because most of mucosal melanotic lesions are painless in their early stages, the diagnosis is unfortunately often delayed until symptoms resulting from ulceration, growth, and/or bleeding are noted. Here, we document the rare case of a malignant rynopharynx melanoma of a 43 year old woman. Its treatment and the pertinent literature are discussed. No complication was recorded in the post-operative period and no further surgery was performed. The follow up showed no recurrence in the same position and with the same characteristics, even after six years. Mucosal melanomas are aggressive tumours and the prognosis in these patients is poor. Clinicians must use treatment strategies that provide functional benefit, so as to maintain quality of life without excessive toxicity.


PERMANENT GENETIC RESOURCES: Isolation and characterization of 12 microsatellite markers in the middle-spotted woodpecker (Dendrocopos medius).
Twelve polymorphic microsatellite loci were developed for Dendrocopos medius. Polymorphism was assessed for 27 individuals from the southwesternmost population of this woodpecker species. The number of alleles per locus ranged from three to seven, with observed heterozygosity values from 0.444 to 0.852. Genotypic frequencies conformed to Hardy-Weinberg equilibrium, and no evidence for linkage disequilibrium was observed. Multilocus genotypes resulting from this set of markers will be useful to determine genetic diversity and differentiation within and among habitat patches inhabited by D. medius. Three of the loci were polymorphic for Picoides articus.


Surrogate data approaches to assess the significance of directed coherence: application to EEG activity propagation.
This paper addresses the topic of evaluating the significance of frequency domain measures of causal coupling in multivariate time series through generation of surrogate data. The considered approaches are the traditional Fourier Transform (FT) algorithm and a new causal FT (CFT) algorithm for surrogate data generation. Both algorithms preserve the FT modulus of the original series; differences are in the phase relationships, that are completely destroyed for FT surrogates and imposed after switching off the link over the considered causal direction for CFT surrogates. The ability of the algorithms to assess causality in the frequency domain was tested using the directed coherence as discriminating parameter. Evaluation on simulated multivariate linear processes and application over multichannel EEG recordings showed that the utilization of CFT surrogates improves specificity of the test for nonzero spectral causality, as FT surrogates may attribute to a direct coupling the presence of indirect connectivity patterns.


Cystic fibrosis: A changing landscape.
Cystic fibrosis is an autosomal recessive genetic disorder that causes a lifetime of debilitating and potentially fatal complications affecting the lungs and other organ systems. Over 1,700 gene mutations that cause this rare disorder have been identified. This article describes the current treatment landscape for adults with CF, including the 2019 FDA approval of a breakthrough triple-drug combination therapy that may significantly improve the quality of life for an estimated 90% of patients with CF.


The impact of ultrasound-based antenatal screening strategies to detect vasa praevia in the United Kingdom: An exploratory study using decision analytic modelling methods.
The objective of this exploratory modelling study was to estimate the effects of second-trimester, ultrasound-based antenatal detection strategies for vasa praevia (VP) in a hypothetical cohort of pregnant women. For this, a decision-analytic tree model was developed covering four discrete detection pathways/strategies: no screening; screening targeted at women undergoing in-vitro fertilisation (IVF); screening targeted at women with low-lying placentas (LLP); screening targeted at women with velamentous cord insertion (VCI) or a bilobed or succenturiate (BL/S) placenta. Main outcome measures were the number of referrals to transvaginal sonography (TVS), diagnosed and undiagnosed cases of VP, overdetected cases of VCI, and VP-associated perinatal mortality. The greatest number of referrals to TVS occurred in the LLP-based (2,083) and VCI-based screening (1,319) pathways. These two pathways also led to the highest proportions of pregnancies diagnosed with VP (VCI-based screening: 552 [78.9% of all pregnancies]; LLP-based: 371 [53.5%]) and the lowest proportions of VP leading to perinatal death (VCI-based screening: 100 [14.2%]; LLP-based: 196 [28.0%]). In contrast, the IVF-based pathway resulted in 66 TVS referrals, 50 VP diagnoses (7.1% of all VP pregnancies), and 368 (52.6%) VP-associated perinatal deaths which was comparable to the no screening pathway (380 [54.3%]). The VCI-based pathway resulted in the greatest detection of VCI (14,238 [99.1%]), followed by the IVF-based pathway (443 [3.1%]); no VCI detection occurred in the LLP-based or no screening pathways. In conclusion, the model results suggest that a targeted LLP-based approach could detect a substantial proportion of VP cases, while avoiding VCI overdetection and requiring minimal changes to current clinical practice. High-quality data is required to explore the clinical and cost-effectiveness of this and other detection strategies further. This is necessary to provide a robust basis for future discussion about routine screening for VP.


Children's perception of visual and auditory ambiguity and its link to executive functions and creativity.
The phenomenon of perceptual bistability provides insights into aspects of perceptual processing not normally accessible to everyday experience. However, most experiments have been conducted in adults, and it is not clear to what extent key aspects of perceptual switching change through development. The current research examined the ability of 6-, 8-, and 10-year-old children (N = 66) to switch between competing percepts of ambiguous visual and auditory stimuli and links between switching rate, executive functions, and creativity. The numbers of switches participants reported in two visual tasks (ambiguous figure and ambiguous structure from motion) and two auditory tasks (verbal transformation and auditory streaming) were measured in three 60-s blocks. In addition, inhibitory control was measured with a Stroop task, set shifting was measured with a verbal fluency task, and creativity was measured with a divergent thinking task. The numbers of perceptual switches increased in all four tasks from 6 to 10 years of age but differed across tasks in that they were higher in the verbal transformation and ambigous structure-from-motion tasks than in the ambigous figure and auditory streaming tasks for all age groups. Although perceptual switching rates differed across tasks, there were predictive relationships between switching rates in some tasks. However, little evidence for the influence of central processes on perceptual switching was found. Overall, the results support the notion that perceptual switching is largely modality and task specific and that this property is already evident when perceptual switching emerges.


How the carotid body works: different strategies and preparations to solve different problems.
This is a review of the different experimental approaches developed to solve the problems in our progress towards a comprehensive understanding of how arterial chemoreceptors operate. An analysis is performed oi the bases, advantages and limits of the following preparations: studies of ventilatory reflexes originated from carotid bodies (CBs) in the entire animal; recordings of CB chemosensory discharges in situ; CB preparations perfused in situ; CB explants in oculo; CB explants in ovo; CB preparations incubated in vitro; CB preparations superfused in vitro; CB preparations perfused and superfused in vitro: CB tissue slices in vitro; cells acutely dissociated from CBs; CB cells in tissue culture; petrosal ganglia superfused in vitro; petrosal ganglion cells in tissue culture; and co-cultures of CB and sensory ganglion cells. A brief historical account is given of the passage from one preparation to the next one. Emphasis is placed on personal experience with the different preparations whenever possible. Examples are given of the importance of selecting the appropriate experimental preparation for solving each particular theoretical problem. In fact, brilliant ideas on how the CB works have been unproductive until finding the adequate experimental approach to explore the validity of such ideas.


Inference from single occasion capture experiments using genetic markers.
Accurate estimation of the size of animal populations is an important task in ecological science. Recent advances in the field of molecular genetics researches allow the use of genetic data to estimate the size of a population from a single capture occasion rather than repeated occasions as in the usual capture-recapture experiments. Estimating the population size using genetic data also has sometimes led to estimates that differ markedly from each other and also from classical capture-recapture estimates. Here, we develop a closed form estimator that uses genetic information to estimate the size of a population consisting of mothers and daughters, focusing on estimating the number of mothers, using data from a single sample. We demonstrate the estimator is consistent and propose a parametric bootstrap to estimate the standard errors. The estimator is evaluated in a simulation study and applied to real data. We also consider maximum likelihood in this setting and discover problems that preclude its general use.


Design and implementation of a virtual world training simulation of ICU first hour handover processes.
Nursing training for an Intensive Care Unit (ICU) is a resource intensive process. High demands are made on staff, students and physical resources. Interactive, 3D computer simulations, known as virtual worlds, are increasingly being used to supplement training regimes in the health sciences; especially in areas such as complex hospital ward processes. Such worlds have been found to be very useful in maximising the utilisation of training resources. Our aim is to design and develop a novel virtual world application for teaching and training Intensive Care nurses in the approach and method for shift handover, to provide an independent, but rigorous approach to teaching these important skills. In this paper we present a virtual world simulator for students to practice key steps in handing over the 24/7 care requirements of intensive care patients during the commencing first hour of a shift. We describe the modelling process to provide a convincing interactive simulation of the handover steps involved. The virtual world provides a practice tool for students to test their analytical skills with scenarios previously provided by simple physical simulations, and live on the job training. Additional educational benefits include facilitation of remote learning, high flexibility in study hours and the automatic recording of a reviewable log from the session. To the best of our knowledge, we believe this is a novel and original application of virtual worlds to an ICU handover process. The major outcome of the work was a virtual world environment for training nurses in the shift handover process, designed and developed for use by postgraduate nurses in training.


Field Measurement of Effective Leaf Area Index using Optical Device in Vegetation Canopy.
Leaf area index (LAI) is an essential canopy variable describing the amount of foliage in an ecosystem. The parameter serves as the interface between green components of plants and the atmosphere, and many physiological processes occur there, primarily photosynthetic uptake, respiration, and transpiration. LAI is also an input parameter for many models involving carbon, water, and the energy cycle. Moreover, ground-based in situ measurements serve as the calibration method for LAI obtained from remote sensing products. Therefore, straightforward indirect optical methods are necessary for making precise and rapid LAI estimates. The methodological approach, advantages, controversies, and future perspectives of the newly developed LP 110 optical device based on the relation between radiation transmitted through the vegetation canopy and canopy gaps were discussed in the protocol. Furthermore, the instrument was compared to the world standard LAI-2200 Plant Canopy Analyzer. The LP 110 enables more rapid and more straightforward processing of data acquired in the field, and it is more affordable than the Plant Canopy Analyzer. The new instrument is characterized by its ease of use for both above- and below-canopy readings due to its greater sensor sensitivity, in-built digital inclinometer, and automatic logging of readings at the correct position. Therefore, the hand-held LP 110 device is a suitable gadget for performing LAI estimation in forestry, ecology, horticulture, and agriculture based on the representative results. Moreover, the same device also enables the user to take accurate measurements of incident photosynthetically active radiation (PAR) intensity.


Automating and Extending Comprehensive Two-Dimensional Gas Chromatography Data Processing by Interfacing Open-Source and Commercial Software.
Comprehensive two-dimensional gas chromatography (GC×GC) is a powerful analytical tool for both nontargeted and targeted analyses. However, there is a need for more integrated workflows for processing and managing the resultant high-complexity datasets. End-to-end workflows for processing GC×GC data are challenging and often require multiple tools or software to process a single dataset. We describe a new approach, which uses an existing underutilized interface within commercial software to integrate free and open-source/external scripts and tools, tailoring the workflow to the needs of the individual researcher within a single software environment. To demonstrate the concept, the interface was successfully used to complete a first-pass alignment on a large-scale GC×GC metabolomics dataset. The analysis was performed by interfacing bespoke and published external algorithms within a commercial software environment to automatically correct the variation in retention times captured by a routine reference standard. Variation in 1tR and 2tR was reduced on average from 8 and 16% CV prealignment to less than 1 and 2% post alignment, respectively. The interface enables automation and creation of new functions and increases the interconnectivity between chemometric tools, providing a window for integrating data-processing software with larger informatics-based data management platforms.


The Doa4 deubiquitinating enzyme is functionally linked to the vacuolar protein-sorting and endocytic pathways.
The Saccharomyces cerevisiae DOA4 gene encodes a deubiquitinating enzyme that is required for rapid degradation of ubiquitin-proteasome pathway substrates. Both genetic and biochemical data suggest that Doa4 acts in this pathway by facilitating ubiquitin recycling from ubiquitinated intermediates targeted to the proteasome. Here we describe the isolation of 12 spontaneous extragenic suppressors of the doa4-1 mutation; these involve seven different genes, six of which were cloned. Surprisingly, all of the cloned DID (Doa4-independent degradation) genes encode components of the vacuolar protein-sorting (Vps) pathway. In particular, all are class E Vps factors, which function in the maturation of a late endosome/prevacuolar compartment into multivesicular bodies that then fuse with the vacuole. Four of the six Did proteins are structurally related, suggesting an overlap in function. In wild-type and several vps strains, Doa4-green fluorescent protein displays a cytoplasmic/nuclear distribution. However, in cells lacking the Vps4/Did6 ATPase, a large fraction of Doa4-green fluorescent protein, like several other Vps factors, concentrates at the late endosome-like class E compartment adjacent to the vacuole. These results suggest an unanticipated connection between protein deubiquitination and endomembrane protein trafficking in which Doa4 acts at the late endosome/prevacuolar compartment to recover ubiquitin from ubiquitinated membrane proteins en route to the vacuole.


Resolution of glucose intolerance in long-term high-fat, high-sucrose-fed mice.
The high-fat, high-sucrose diet (HFSD)-fed C57Bl/6 mouse is a widely used model of prediabetes. However, studies typically implement a relatively short dietary intervention lasting between 4 and 16 weeks; as a result, little is known about how a long-term HFSD influences the metabolic profile of these mice. Therefore, the aim of this investigation was to examine the effects of consuming a HFSD for 42 weeks on the development of hyperinsulinaemia and glucose intolerance in male C57Bl/6 mice. Two cohorts of HFSD mice were studied at independent institutes and they underwent an oral glucose tolerance test (OGTT) with measures of plasma insulin and free fatty acids (FFA). Age-matched chow-fed control mice were also studied. The HFSD-fed mice were hyperinsulinaemic and grossly obese, being over 25 g heavier than chow-fed mice, which was due to a marked expansion of subcutaneous adipose tissue. This was associated with a 3-fold increase in liver lipid content. Glucose tolerance, however, was either the same or better than control mice due to the preservation of glucose disposal as revealed by a dynamic stable isotope-labelled OGTT. In addition, plasma FFAs were suppressed to lower levels in HFSD mice during the OGTT. In conclusion, we have made the paradoxical observation that long-term HFSD feeding results in the resolution of glucose intolerance in the C57Bl/6 mouse. Mechanistically, we propose that the gross expansion of subcutaneous adipose tissue increases the glucose disposal capacity of the HFSD-fed mouse, which overcomes the prevailing insulin resistance to improve glucose tolerance.


[CT-stereotaxic punctures, aspiration and drainage of deep space-occupying processes in the brain (cystic tumors, hematomas and abscesses)].
The article analyses the clinical material concerning 68 patients with various deeply-seated space-occupying processes of the brain (cystic tumors, multiple abscesses, intracerebral hematomas) which were subjected to stereotaxic aspiration on the basis of the findings of computed tomography (CT-stereotaxic punctures). A significant and immediate effect with rapid regression of local and general cerebral symptoms was recorded in 38 patients with cystic intracerebral tumors and craniopharyngiomas. All the patients tolerated the operation well and in none of them were complications encountered. The precision, reliability and safety of CT-stereotaxic punctures--is the basis for such operations as intra-tissue beta-therapy of craniopharyngiomas with radiocolloids, evacuation of intracerebral hematomas, and drainage of multiple brain abscesses. Being an obvious palliative surgical measure, CT-stereotaxic aspiration of deeply-seated cystic tumors is a method of choice in cases in which a direct operation is contraindicated and in patients who are in a grave condition.


Pediatric Idiopathic Intracranial Hypertension Presenting With Sensorineural Hearing Loss.
To present the rare case of a young boy with idiopathic intracranial hypertension presenting with bilateral sensorineural hearing loss developing over several months. This was accompanied by headaches, otalgia, tinnitus, and vertigo. Furthermore, we aim to provide a concise review on this matter, as this report represents the second case in literature of pediatric idiopathic intracranial hypertension presenting with hearing loss. Workup of a 9-year-old boy with bilateral sensorineural hearing loss, including (among others) physical examination, audiometry, diagnostic imaging, and lumbar puncture. Physical examination including fundoscopy as well as imaging showed no abnormalities. At presentation, pure tone audiometry revealed bone conduction thresholds of about 30 dB HL in both ears. Two months later, this declined to about 35 dB HL in both ears. Lumbar puncture revealed an increased intracranial pressure. The boy was thus diagnosed with idiopathic intracranial hypertension. After the lumbar puncture, the otological complaints gradually resolved, and the hearing normalized (bone conduction thresholds of 0-5 dB HL). Although rare, sensorineural hearing loss in the pediatric population together with otalgia, tinnitus, and vertigo can be due to idiopathic intracranial hypertension and as such can be reversible.


Decreased acute rejection in kidney transplant recipients is associated with decreased chronic rejection.
To determine whether a recent decrease in the rate of acute rejection after kidney transplantation was associated with a decrease in the rate of chronic rejection. Single-institution and multicenter retrospective analyses have identified acute rejection episodes as the major risk factor for chronic rejection after kidney transplantation. However, to date, no study has shown that a decrease in the rate of acute rejection leads to a decrease in the rate of chronic rejection. The authors studied patient populations who underwent transplants at a single center during two eras (1984-1987 and 1991-1994) to determine the rate of biopsy-proven acute rejection, the rate of biopsy-proven chronic rejection, and the graft half-life. Recipients who underwent transplantation in era 2 had a decreased rate of biopsy-proven acute rejection compared with era 1 (p < 0.05). This decrease was associated with a decreased rate of biopsy-proven chronic rejection for both cadaver (p = 0.0001) and living donor (p = 0.08) recipients. A trend was observed toward increased graft half-life in era 2 (p = NS). Development of immunosuppressive protocols that decrease the rate of acute rejection should lower the rate of chronic rejection and improve long-term graft survival.


Microbial Leakage at Morse Taper Conometric Prosthetic Connection: An In Vitro Investigation.
To evaluate in vitro the sealing capability at the prosthetic connection interface of 2 conometric systems. Two conometric systems with the same design and different material were used, for a total of 24 samples. Each sample was assembled by a tapered abutment and respective coping. In group A, the copings were made of gold, whereas in group B they were made of PEEK. Three μL of mix bacterial suspension (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and Fusobacterium nucleatum species) was inoculated into the abutment screw hole, and the coping was inserted on the abutment. Samples were immersed into culture tubes and incubated for 24, 48, and 72 hours into anaerobic conditions. Visual evaluation of turbidity was performed at each time point. Qualitative-quantitative assessment using real-time polymerase chain reaction was performed at 72 hours. Any difference between the groups was checked by means of Fisher exact test. Microbial leakage occurred in both groups, and there was no statistically significant difference between groups. Microbial concentration resulted in a presence inferior to 1 × 10 copies/μL in all positive assemblies. Because of the low bacterial count, it can be concluded that a minimal bacterial infiltration may be allowed by conometric interfaces for prosthetic connection.


Rhodium-Catalyzed Asymmetric Functionalization of Quinoxalinium Salts.
We report a rhodium-catalyzed asymmetric addition of aryl and alkenyl boronic acids to quinoxalinium salts that generates dihydroquinoxalines with high enantioselectivity. Functionalization of the reaction products, dihydroquinoxaline, allows the preparation of tetrahydroquinoxalines with various substitution patterns.


Association of IGF-I and IGF-II with myofiber regeneration in vivo.
This study examined expression of insulinlike growth factor (IGF) in the myofibers and nonmyofibrillar structures of murine soleus muscle following contraction-induced damage. Identifying the cellular sources of this myogenic growth factor could improve muscle rehabilitation strategies. Immunohistochemical analysis of muscle sections indicated that the number of myofibers expressing both IGF-I and IGF-II increased significantly at 4, 7, and 10 days following injury, compared with control. Muscle spindles and vascular tissue expressed only IGF-II, and staining intensity did not change following injury. The number of fibers expressing developmental myosin heavy chain increased significantly at 7 and 10 days postinjury, and these usually coexpressed IGF. No IGF-specific staining of interstitial/inflammatory cells was observed. Therefore, expression of IGF after mechanically induced fiber damage occurs exclusively within regenerating fibers without supplemental delivery of IGF to the tissue by inflammatory cells or changes in constitutive expression of IGF-II in vascular tissue.


Prevention of Recurrent Childhood Caries with Probiotic Supplements: A Randomized Controlled Trial with a 12-Month Follow-Up.
The aim of this study was to evaluate the effect of drops containing probiotic bacteria on the recurrence of dental caries in preschool children. The study employed a randomized, placebo-controlled, double-blinded design with two parallel arms. 38 preschool children were enrolled after comprehensive restorative treatment under general anesthesia or conscious sedation (baseline), and they were followed up after 6 and 12 months. Parents of children in the test group were instructed to give 5 daily drops containing two strains of Limosilactobacillus reuteri (DSM 17938 and ATCC PTA 5289) at bedtime. The placebo drops were identically composed but lacked bacteria. The duration of the intervention was 12 months. The primary endpoint was recurrence of new caries lesions on subject level (yes/no), and secondary endpoints were presence of dental plaque and gingivitis. We found high rate of recurrent moderate and extensive lesions after 12 months (67%) but there were no significant differences between the groups. We observed no beneficial effects on dental plaque or gingival inflammation. The findings were however uncertain and inconclusive due to lack of power, a consequence of the COVID-19 pandemic. ClinTrials.gov Identifier: (NCT04929340), June 18, 2021; retrospectively registered.


Intestinal parasites at the Late Bronze Age settlement of Must Farm, in the fens of East Anglia, UK (9th century B.C.E.).
Little is known about the types of intestinal parasites that infected people living in prehistoric Britain. The Late Bronze Age archaeological site of Must Farm was a pile-dwelling settlement located in a wetland, consisting of stilted timber structures constructed over a slow-moving freshwater channel. At excavation, sediment samples were collected from occupation deposits around the timber structures. Fifteen coprolites were also hand-recovered from the occupation deposits; four were identified as human and seven as canine, using fecal lipid biomarkers. Digital light microscopy was used to identify preserved helminth eggs in the sediment and coprolites. Eggs of fish tapeworm (Diphyllobothrium latum and Diphyllobothrium dendriticum), Echinostoma sp., giant kidney worm (Dioctophyma renale), probable pig whipworm (Trichuris suis) and Capillaria sp. were found. This is the earliest evidence for fish tapeworm, Echinostoma worm, Capillaria worm and the giant kidney worm so far identified in Britain. It appears that the wetland environment of the settlement contributed to establishing parasite diversity and put the inhabitants at risk of infection by helminth species spread by eating raw fish, frogs or molluscs that flourish in freshwater aquatic environments, conversely the wetland may also have protected them from infection by certain geohelminths.


Is the "South Asian Phenotype" Unique to South Asians?: Comparing Cardiometabolic Risk Factors in the CARRS and NHANES Studies.
In the context of rising obesity in South Asia, it is unclear whether the "South Asian phenotype"(described as high glucose, low high-density lipoprotein cholesterol, and high triglycerides at normal ranges of body weight) continues to be disproportionately exhibited by contemporary South Asians relative to other race/ethnic groups. We assessed the distinctiveness of the South Asian cardiometabolic profile by comparing the prevalence of combined high glucose, high triglycerides, and low high-density lipoprotein cholesterol (combined dysglycemia and dyslipidemia) in resident South Asians with 4 race/ethnic groups in the United States (Asians, black persons, Hispanics, and white persons) overall and by body mass index (BMI) category. South Asian data were from the 2010 to 2011 Center for Cardiometabolic Risk Reduction in South Asia Study, representative of Chennai and New Delhi, India and Karachi, Pakistan. U.S. data were from the 2011 to 2012 National Health and Nutrition Examination Survey, representative of the U.S. Combined dysglycemia and dyslipidemia was defined as fasting blood glucose ≥126 mg/dl and triglyceride/high-density lipoprotein cholesterol ratio >4. Logistic regression was used to estimate the relative odds and 95% confidence intervals of combined dysglycemia and dyslipidemia associated with each race/ethnic group (referent, U.S. white persons). Models were estimated among adults aged 20 to 79 years by sex and BMI category and accounted for age, education, and tobacco use. Data from 8,448 resident South Asians, 274 U.S. Asians, 404 U.S. black persons, 308 U.S. Hispanics, and 703 U.S. white persons without previously known diabetes were analyzed. In the normal body weight range of BMI 18.5 to 24.9 kg/m(2), the prevalence of combined dysglycemia and dyslipidemia among men and women, respectively, was 33% and 11% in resident South Asians, 15% and 1% in U.S. Asians, 5% and 2% in U.S. black persons, 11% and 2% in U.S. Hispanics, and 8% and 2% in U.S. white persons. Compared with U.S. whites persons, South Asians were more likely to present with combined dysglycemia and dyslipidemia at all categories of BMI for men and at BMI 18.5 to 29.9 for women in adjusted models. The most pronounced difference between South Asians and U.S. white persons was observed at normal weight (adjusted odds ratio: 4.98; 95% confidence interval: 2.46 to 10.07 for men) (adjusted odds ratio: 9.09; 95% confidence interval: 2.48 to 33.29 for women). Between 8% and 15% of U.S. men and 1% and 2% of U.S. women of diverse race/ethnic backgrounds exhibited dysglycemia and dyslipidemia at levels of body weight considered "healthy," consistent with the cardiometabolic profile described as the "South Asian Phenotype." Urban South Asians, however, were 5 to 9 times more likely to exhibit dysglycemia and dyslipidemia in the "healthy" BMI range compared with any other U.S. race/ethnic group.


Role of nitrosative stress in the pathogenesis of diabetic vascular dysfunction.
Here we overview the role of reactive nitrogen species (nitrosative stress) and associated pathways in the pathogenesis of diabetic vascular complications. Increased extracellular glucose concentration, a principal feature of diabetes mellitus, induces a dysregulation of reactive oxygen and nitrogen generating pathways. These processes lead to a loss of the vascular endothelium to produce biologically active nitric oxide (NO), which impairs vascular relaxations. Mitochondria play a crucial role in this process: endothelial cells placed in increase extracellular glucose respond with a marked increase in mitochondrial superoxide formation. Superoxide, when combining with NO generated by the endothelial cells (produced by the endothelial isoform of NO synthase), leads to the formation of peroxynitrite, a cytotoxic oxidant. Reactive oxygen and nitrogen species trigger endothelial cell dysfunction through a multitude of mechanisms including substrate depletion and uncoupling of endothelial isoform of NO synthase. Another pathomechanism involves DNA strand breakage and activation of the nuclear enzyme poly(ADP-ribose) polymerase (PARP). PARP-mediated poly(ADP-ribosyl)ation and inhibition of glyceraldehyde-3-phosphate dehydrogenase importantly contributes to the development of diabetic vascular complications: it induces activation of multiple pathways of injury including activation of nuclear factor kappa B, activation of protein kinase C and generation of intracellular advanced glycation end products. Reactive species generation and PARP play key roles in the pathogenesis of 'glucose memory' and in the development of injury in endothelial cells exposed to alternating high/low glucose concentrations.


The immunocytochemical localization of connexin 36 at rod and cone gap junctions in the guinea pig retina.
Connexin 36 (Cx36) is a channel-forming protein found in the membranes of apposed cells, forming the hexameric hemichannels of intercellular gap junction channels. It localizes to certain neurons in various regions of the brain including the retina. We characterized the expression pattern of neuronal Cx36 in the guinea pig retina by immunocytochemistry using specific antisera against Cx36 and green/red cone opsin or recoverin. Strong Cx36 immunoreactivity was visible in the ON sublamina of the inner plexiform layer and in the outer plexiform layer, as punctate labelling patterns. Double-labelling experiments with antibody directed against Cx36 and green/red cone opsin or recoverin showed that strong clustered Cx36 immunoreactivity localized to the axon terminals of cone or close to rod photoreceptors. By electron microscopy, Cx36 immunoreactivity was visible in the gap junctions as well as in the cytoplasmic matrices of both sides of cone photoreceptors. In the gap junctions between cone and rod photoreceptors, Cx36 immunoreactivity was only visible in the cytoplasmic matrices of cone photoreceptors. These results clearly indicate that Cx36 forms homologous gap junctions between neighbouring cone photoreceptors, and forms heterologous gap junctions between cone and rod photoreceptors in guinea pig retina. This focal location of Cx36 at the terminals of the photoreceptor suggests that rod photoreceptors can transmit rod signals to the pedicle of a neighbouring cone photoreceptor via Cx36, and that the cone in turn signals to corresponding ganglion cells via ON and OFF cone bipolar cells.


[Alteration of white rats brain tissue inducted by assessment of silver nanocomposite incapsulated in polymer matrix].
The paper present experimental materials of intragastric administration of silver nanoparticles encapsulated in polymer matrix of arabinogalactan by white outbred male rats. Animals were injected "pure" arabinogalactan and colloid silver solution containing silver macroform separately for comparison. Research provided data about status of brain tissue at the impact of these substances on organism. Histological analysis revealed the presence of a pathological process, character and intensity of which varied depending on the type of injected material. Pathological process under the influence of silver-arabinogalactan characterized by appearance in brain tissue of perivascular edema and development of acute inflammation in formation of glial scars, swelling of vascular bundles in sum.


Enhancement of mucosal innate and adaptive immunity following intranasal immunization of mice with a bovine adenoviral vector.
Nonhuman adenoviral (AdV) gene delivery platforms have significant value due to their ability to elude preexisting AdV vector immunity in most individuals. Previously, we have demonstrated that intranasal (IN) immunization of mice with BAd-H5HA, a bovine AdV type 3 (BAdV3) vector expressing H5N1 influenza virus hemagglutinin (HA), resulted in enhanced humoral and cell-mediated immune responses. The BAd-H5HA IN immunization resulted in complete protection following the challenge with an antigenically distinct H5N1 virus compared to the mouse group similarly immunized with HAd-H5HA, a human AdV type 5 (HAdV5) vector expressing HA. Here, we attempted to determine the activation of innate immune responses in the lungs of mice inoculated intranasally with BAd-H5HA compared to the HAd-H5HA-inoculated group. RNA-Seq analyses of the lung tissues revealed differential expression (DE) of genes involved in innate and adaptive immunity in animals immunized with BAd-H5HA. The top ten enhanced genes were verified by RT-PCR. Consistently, there were transient increases in the levels of cytokines (IL-1α, IL-1β, IL-5, TNF- α, LIF, IL-17, G-CSF, MIP-1β, MCP-1, MIP-2, and GM-CSF) and toll-like receptors in the lungs of the group inoculated with BAdV vectors compared to that of the HAdV vector group. These results demonstrate that the BAdV vectors induce enhanced innate and adaptive immunity-related factors compared to HAdV vectors in mice. Thus, the BAdV vector platform could be an excellent gene delivery system for recombinant vaccines and cancer immunotherapy.


Immunophenotypic analysis of myelodysplastic syndromes.
In contrast with hematologic malignancies in which the value of immunophenotypic studies is well established, information on the immunophenotypic characteristics of myelodysplastic syndromes (MDS) is scanty. The main goal of the present study was to explore the immunophenotypic differences between patients with MDS and normal individuals, including changes in distribution of cell lineages as well as phenotypic aberrations and blockades in cell maturation pathways. In MDS the proportion of bone marrow CD34+ cells was higher than in normal patients but the most immature progenitors (CD34+CD38-) were less represented. By contrast the proportion of myelomonocytic CD34+ cells was greater than in normal individuals, translating into an increased myeloid/non-myeloid CD34+ hematopoietic progenitor cell ratio. This suggests that in MDS, the majority of CD34+ cells are already committed to the myeloid lineage. Upon analyzing the granulo-monocytic differentiation pathway, MDS patients showed an increased proportion of monocytic cells with a decreased percentage of cells of neutrophil lineage, leading to a lower neutrophil/monocytic cell ratio. Maturational arrests in the monocytic but not in the neutrophil differentiation pathway were observed. In refractory anemia with excess blasts in transformation (RAEB-t) such blockades mainly occurred during the earliest stages of differentiation but in the other MDS subtypes they occurred in later stages. Phenotypic aberrations occurred in 90% of patients and a high proportion of cases showed >=2 aberrations. In summary, our results show that, in addition to an abnormal distribution of the bone marrow cell compartment, MDS patients frequently show aberrant phenotypes and maturational arrests. Some of these features may help in cases in which the diagnosis of MDS is questionable.


NMR imaging in the recognition of giant intracranial aneurysms.
Most giant aneurysms present clinically as intracranial mass lesions. Many contain an organized lamellated thrombus with only a small eccentric lumen remaining that can be opacified at angiography. Four patients with such aneurysms were studied using flow-dependent nuclear magnetic resonance scanning sequences. The aneurysms show a smooth rounded margin, and the thrombus within is clearly distinguishable from the blood flowing through the patent lumen, which gives a low signal. By using different scan sequences it is possible to prove that the area of low signal is indeed fluid in rapid motion, thus confirming the diagnosis.