Urethral catheters: can we reduce use?
Indwelling urinary catheters are the main cause of healthcare-associated urinary tract infections. It can be expected that reduction of the use of urinary catheters will lead to decreased numbers of urinary tract infection. The efficacy of an intervention programme to improve adherence to recommendations to reduce the use of urethral catheters was studied in a before-after comparison in ten Dutch hospitals. The programme detected barriers and facilitators and each individual facility was supported with developing their own intervention strategy. Outcome was evaluated by the prevalence of catheters, alternatives such as diapers, numbers of urinary tract infections, the percentage of correct indications and the duration of catheterization. The costs of the implementation as well as the catheterization were evaluated. Of a population of 16,495 hospitalized patients 3335 patients of whom 2943 were evaluable for the study, had a urethral catheter. The prevalence of urethral catheters decreased insignificantly in neurology (OR 0.93; 95% CI 0.77 - 1.13) and internal medicine wards (OR 0.97; 95% CI 0.83 - 1.13), decreased significantly in surgical wards (OR 0.84; 95% CI 0.75 - 0.96), but increased significantly in intensive care (IC) and coronary care (CC) units (OR 1.48; 95% CI 1.01 - 2.17). The use of alternatives was limited and remained so after the intervention. Duration of catheterization decreased insignificantly in IC/CC units (ratio after/before 0.95; 95% CI 0.78 - 1.16) and neurology (ratio 0.97; 95% CI 0.80 - 1.18) and significantly in internal medicine (ratio 0.81; 95% CI 0.69 - 0.96) and surgery wards (ratio 0.80; 95% CI 0.71 - 0.90). The percentage of correct indications on the day of inclusion increased from 50 to 67% (p < 0.0001). The prevalence of urinary tract infections in catheterized patients did not change. The mean cost saved per 100 patients was € 537. Targeted implementation of recommendations from an existing guideline can lead to better adherence and cost savings. Especially, hospitals which use a lot of urethral catheters or where catheterization is prolonged, can expect important improvements.


Knockdown of caveolin-1 by antisense oligonucleotides impairs angiogenesis in vitro and in vivo.
Knock-out of the gene coding for caveolin-1, the main organizer of caveolae, has not yet been performed. We devised a strategy to knock-down caveolin-1 gene expression using antisense oligodeoxynucleotides (ODNs). Seven ODNs, covering different regions of caveolin-1 mRNA, were screened by Western blot analysis of caveolin-1 levels. The most active and specific was found to reduce caveolin-1 protein levels by 70% at 1 microM concentration and its action, as demonstrated by a marked reduction (about 50%) in caveolin-1 mRNA levels, was due to a true antisense mechanism. In HUVEC treated with the active ODN, caveolae were undetectable by confocal and electron microscopy, while their number was not affected when cells were treated with a scrambled ODN. Using the fibrin gel 3 D angiogenesis test we established that the active (but not the scrambled) ODN strongly suppressed capillary-like tube formation. Moreover, an antisense tailored against chicken caveolin-1 mRNA, when tested using the chorio-allantoic membrane technique, dramatically reduced vessel formation at doses (10-20 microg) under which control ODNs were ineffective and devoid of toxicity. Thus, it is likely that caveolin-1 down regulation, followed by caveolae disruption, impairs angiogenesis in vitro and in vivo.


Effects of soybean isoflavones on the growth performance, intestinal morphology and antioxidative properties in pigs.
Soybean meal is rich in soybean isoflavones, which exhibit antioxidant, anti-inflammatory, antiviral and anticancer functions in humans and animals. This study was conducted to investigate the effects of soybean isoflavones on the growth performance, intestinal morphology and antioxidative properties in pigs. A total of 72 weaned piglets (7.45 ± 0.13 kg; 36 males and 36 females) were allocated into three treatments and fed corn-soybean meal (C-SBM), corn-soy protein concentrate (C-SPC) or C-SPC supplemented with equal levels of the isoflavones found in the C-SBM diet (C-SPC + ISF) for a 72-day trial. Each treatment had six replicates and four piglets per replicate, half male and half female. On day 42, one male pig from each replicate was selected and euthanized to collect intestinal samples. The results showed that compared to pigs fed the C-SPC diet, pigs fed the C-SBM and C-SPC + ISF diets had higher BW on day 72 (P < 0.05); pigs fed the C-SBM diet had significantly higher average daily gain (ADG) during days 14 to 28 (P < 0.05), with C-SPC + ISF being intermediate; pigs fed the C-SBM diet tended to have higher ADG during days 42 to 72 (P = 0.063), while pigs fed the C-SPC + ISF diet had significantly higher ADG during days 42 to 72 (P < 0.05). Moreover, compared to pigs fed the C-SPC diet, pigs fed the C-SBM diet tended to have greater villus height (P = 0.092), while pigs fed the C-SPC + ISF diet had significantly greater villus height (P < 0.05); pigs fed the C-SBM and C-SPC + ISF diets had significantly increased villus height-to-crypt depth ratio (P < 0.05). Compared with the C-SPC diet, dietary C-SPC + ISF tended to increase plasma superoxide dismutase activity on days 28 (P = 0.085) and 42 (P = 0.075) and reduce plasma malondialdehyde (MDA) content on day 42 (P = 0.089), as well as significantly decreased jejunal mucosa MDA content on day 42 (P < 0.05). However, no significant difference in the expression of tight junction genes among the three groups was found (P > 0.05). In conclusion, our results suggest that a long-term exposure to soybean isoflavones enhances the growth performance, protects the intestinal morphology and improves the antioxidative properties in pigs.


National and Local Strategies in the Netherlands for Obesity Prevention and Management in Children and Adolescents.
As in many other countries around the world, the Netherlands has a high prevalence of overweight and obesity in children. About 1 in 7 of children aged 2-19 years has either overweight or obesity. In this paper the national and local activities aimed at the prevention and management of obesity in children and adolescents in the Netherlands are reviewed. It is recommended to, nationally as well as locally, take an integrated-systems approach that tackles the obesogenic food environment as well as upstream and downstream determinants of obesity. Efforts should take a life course approach and be focused on promoting obesity prevention as well as improving the management of children who already have obesity. The national policies in the Netherlands rely heavily on self-regulation by stakeholders such as supermarkets, restaurants, and the food industry. Local policies and actions such as the whole-systems approach in Amsterdam are promising. Future directions include development of tools for the operationalization and evaluation of local systems approaches. Regulation by national and local governments is necessary to ensure a healthy food environment for children and their families, but health policies require intersectoral action. Key Messages: In the Netherlands many policies are in place or under development, especially at the municipal level (e.g., in Amsterdam), but more substantial action is urgently required.


The Clinical Utility of the Vulvar Pain Assessment Questionnaire: A Pilot Study.
The aim of the study was to document treatment-seeking experiences of women with chronic vulvar pain, comfort communicating about pain, and test the clinical utility of the screening version of the Vulvar Pain Assessment Questionnaire, screening version (VPAQscreen). Patients scheduled for an appointment with the Program in Vulvar Health at Oregon Health and Science University were invited to complete the VPAQscreen and answer descriptive questions about previous treatment-seeking experiences and communication with health care providers. Clinicians provided provisional diagnoses based on VPAQscreen summaries, final diagnoses based on gynecological examination, and commented on alignment with clinical observations. Patients gave feedback on the accuracy and helpfulness of the VPAQscreen summary, characteristics of the questions asked, and whether their comfort communicating increased. Participants reported previously seeing approximately 5 medical doctors and 2 other health care providers and perceived them as lacking knowledge of vulvar pain syndromes. Providers indicated that VPAQscreen summaries aligned with clinical presentations and suggested provisional diagnoses with more than 80% accuracy. Participants reported that VPAQscreen summaries were helpful and accurate in summarizing symptoms. Most reported that the number, range, and readability of VPAQscreen questions were good or excellent. More than half reported that completing the VPAQscreen increased comfort when speaking with their Oregon Health and Science University physician. Patients with vulvar pain often endure a lengthy process of consulting multiple clinicians before securing care. The VPAQscreen was more than 80% accurate in predicting diagnosis at this specialty clinic and was useful in assisting patients with expressing symptoms. The applicability of the VPAQscreen in general practice is unknown, although it shows promise.


Sequence heterogeneity of NS5A and core proteins of hepatitis C virus and virological responses to pegylated-interferon/ribavirin combination therapy.
Both host and viral factors have been implicated in influencing the response to pegylated-interferon/ribavirin (PEG-IFN/RBV) therapy for hepatitis C virus (HCV) infection. Among the viral factors, sequence heterogeneity within NS5A and core regions has been proposed. This study aimed to clarify the relationship between virological responses to PEG-IFN/RBV therapy and sequence heterogeneity within NS5A, including the IFN/RBV resistance-determining region (IRRDR), the interferon sensitivity-determining region (ISDR) and the core region. Pretreatment sequences of NS5A and the core regions were analyzed in 57 HCV-1b-infected patients who were to be treated with PEG-IFN/RBV. Of 40 patients infected with HCV having an IRRDR with four or more mutations (IRRDR ≥ 4), 28 (70%) patients achieved a sustained virological response (SVR). On the other hand, only 4 (24%) of 17 patients infected with HCV having an IRRDR with three or fewer mutations (IRRDR ≤ 3) achieved a SVR (P = 0.001). Similarly, 22 (71%) of 31 patients infected with HCV and having an ISDR with one or more mutations (ISDR ≥ 1) achieved a SVR while 10 (38%) of 26 patients infected with HCV and having an ISDR without any mutations (ISDR = 0) achieved a SVR (P = 0.014). As for the core region, there was significant correlation between a single mutation at position 70 (Gln(70) ) and non-SVR (P = 0.02). Notably, Gln(70) was more prominently associated with the null response (P = 0.0007). In conclusion, sequence heterogeneity within the IRRDR and ISDR, and a single point mutation at position 70 of the core region of HCV-1b are likely to be correlated with virological responses to PEG-IFN/RBV therapy.


Comparison of the efficacy of anti-diabetic medications as add-on to metformin in type 2 diabetes mellitus from a real-world database.
Metformin is recommended as a first-line drug in the guidelines of the treatment for type 2 diabetes mellitus. However, high-quality evidence from clinical trials directly comparing the degree of hypoglycemic effect of combination therapy of metformin and a hypoglycemic agent with a different mechanism of action with that of monotherapy of a hypoglycemic drug is lacking. We aimed to examine whether combination therapy of hypoglycemic agents with metformin showed antagonism, addition, or synergism compared to monotherapy with hypoglycemic agents other than metformin regarding hemoglobin A1c levels. This retrospective cohort study used a medical information database in Japan. Non-insulin anti-hyperglycemic agents with different mechanisms of action were classified into eight drug classes. A monotherapy cohort and a combination therapy added to the metformin cohort were defined. The change in hemoglobin A1c levels was evaluated to compare the treatment effect between the cohorts. A total of 13,359 patients with type 2 diabetes mellitus in the monotherapy cohort and 1,064 in the metformin combination therapy cohort were identified. A comparison of the change from baseline HbA1c level by drug class between the two cohorts showed a similar trend. Among those treated with dipeptidyl peptidase-4 inhibitor and sodium-glucose co-transporter-2 inhibitor, no clinically significant difference was observed between the two cohorts (0.00% and -0.07% for unadjusted, 0.15% and -0.03% for propensity score matching-adjusted, and 0.09% and -0.01% for inverse probability treatment weighting-adjusted analysis). According to the results of this study, the effect of dipeptidyl peptidase-4 inhibitor or sodium-glucose co-transporter-2 inhibitor added to metformin seems to be additive with respect to the reduction in hemoglobin A1c.


A survey of urban climate change experiments in 100 cities.
Cities are key sites where climate change is being addressed. Previous research has largely overlooked the multiplicity of climate change responses emerging outside formal contexts of decision-making and led by actors other than municipal governments. Moreover, existing research has largely focused on case studies of climate change mitigation in developed economies. The objective of this paper is to uncover the heterogeneous mix of actors, settings, governance arrangements and technologies involved in the governance of climate change in cities in different parts of the world. The paper focuses on urban climate change governance as a process of experimentation. Climate change experiments are presented here as interventions to try out new ideas and methods in the context of future uncertainties. They serve to understand how interventions work in practice, in new contexts where they are thought of as innovative. To study experimentation, the paper presents evidence from the analysis of a database of 627 urban climate change experiments in a sample of 100 global cities. The analysis suggests that, since 2005, experimentation is a feature of urban responses to climate change across different world regions and multiple sectors. Although experimentation does not appear to be related to particular kinds of urban economic and social conditions, some of its core features are visible. For example, experimentation tends to focus on energy. Also, both social and technical forms of experimentation are visible, but technical experimentation is more common in urban infrastructure systems. While municipal governments have a critical role in climate change experimentation, they often act alongside other actors and in a variety of forms of partnership. These findings point at experimentation as a key tool to open up new political spaces for governing climate change in the city.


Chemical compositions by using LC-MS/MS and GC-MS and biological activities of Sedum sediforme (Jacq.) Pau.
In this research, the chemical composition and biological activities of various extracts obtained from whole parts of Sedum sediforme (Jacq.) Pau were compared. The amounts of total phenolic and flavonoid components in crude extracts were determined by expression as pyrocatechol and quercetin equivalents, respectively. All of the extracts (petroleum ether, acetone, methanol, and water) obtained from S. sediforme showed strong antioxidant activity in four tested methods. Particularly, the IC50 values of the methanol extract, which was the richest in terms of total phenolic and flavonoid contents, were found to be lower than those of α-tocopherol and BHT in β-carotene bleaching (9.78 ± 0.06 μg/mL), DPPH free radical scavenging (9.07 ± 0.07 μg/mL), and ABTS cation radical scavenging (5.87 ± 0.03 μg/mL) methods. Furthermore, the methanol extract of S. sediforme showed higher inhibition activity than galanthamine against acetyl- and butyryl-cholinesterase enzymes. Also, acetone and methanol extracts exhibited moderate antimicrobial activity against Candida albicans. The main constituents of fatty acid and essential oil were identified as palmitic acid (C16:0) (28.8%) and α-selinene (20.4%), respectively, by GC-MS. In the methanol extract of S. sediforme, quercetin, rutin, naringenin, and protocatechuic, p-coumaric, caffeic, and chlorogenic acids were detected and quantified by LC-MS/MS. Results of the current study showed that the methanol extract of S. sediforme may also be used as a food supplement.


A core-shell-structured molecularly imprinted polymer on upconverting nanoparticles for selective and sensitive fluorescence sensing of sulfamethazine.
A core-shell structured molecularly imprinted polymer on upconverting nanoparticles (UCNPs@MIP) was synthesized for the fluorescence (FL) sensing of sulfamethazine (SMZ). Hexagonal UCNPs were synthesized by the solvothermal method, then coated with a thin silica shell and modified with vinyl groups. Finally, surface polymerization was initiated among the vinyl groups, the functional monomers and cross-linking agents by the initiator. The MIP synthesized by this procedure was anchored on the surface of UCNPs, possessed better site accessibility and lower transfer resistance for the target molecule compared to bulk imprinted materials. The obtained UCNPs@MIP showed good binding capacity, fast response, high selectivity and specificity to the SMZ. The FL intensity of the UCNPs@MIP decreased sensitively with the increasing concentration of SMZ in the range of 50-700 ng mL(-1), the detection limit was 34 ng mL(-1) (S/N = 3). The UCNPs@MIP was successfully applied to the detection of SMZ in chicken samples. Thanks to the unique near-infrared (NIR) excitation nature of UCNPs, the chicken meat only needed some simple extraction procedures before FL detection, no complex purifications were required. The average recoveries ranged from 96.01% to 98.90%, with relative standard deviations (RSDs) below 4.5%. This work offers a novel sensing system that combined the advantages of upconverting nanotechnology and molecularly imprinted technology.


High Voltages in Sliding Water Drops.
Water drops on insulating hydrophobic substrates can generate electric potentials of kilovolts upon sliding for a few centimeters. We show that the drop saturation voltage corresponds to an amplified value of the solid-liquid surface potential at the substrate. The amplification is given by the substrate geometry, the drop and substrate dielectric properties, and the Debye length within the liquid. Next to enabling an easy and low-cost way to measure surface- and zeta- potentials, the high drop voltages have implications for energy harvesting, droplet microfluidics, and electrostatic discharge protection.


Behavioral adjustment moderates the effect of neuroticism on brain volume relative to intracranial volume.
The present study examined whether the effect of neuroticism on brain structure is moderated by behavioral adjustment. Neuroticism is widely thought to be harmful to health. However, recent work using proinflammatory biomarkers showed that this effect depends on behavioral adjustment, the willingness and ability to adjust and cope with environmental contingencies, such as different opinions of others or unpredictable life situations. Here, we sought to extend this observation to "brain health" by testing total brain volume (TBV). Using a community sample of 125 Americans, we examined structural magnetic resonance imaging of the brain and quantified TBV. We tested whether the effect of neuroticism on TBV was moderated by behavioral adjustment, net of intracranial volume, age, sex, educational achievement, and race. Behavioral adjustment significantly moderated the effect of neuroticism on TBV, such that neuroticism was associated with lower TBV only when behavioral adjustment was low. There was no such effect when behavioral adjustment was high. The present findings suggest that neuroticism is not debilitating to those who constructively cope with stress. Implications are further discussed.


A computationally efficient framework for the simulation of cardiac perfusion using a multi-compartment Darcy porous-media flow model.
We present a method to efficiently simulate coronary perfusion in subject-specific models of the heart within clinically relevant time frames. Perfusion is modelled as a Darcy porous-media flow, where the permeability tensor is derived from homogenization of an explicit anatomical representation of the vasculature. To account for the disparity in length scales present in the vascular network, in this study, this approach is further refined through the implementation of a multi-compartment medium where each compartment encapsulates the spatial scales in a certain range by using an effective permeability tensor. Neighbouring compartments then communicate through distributed sources and sinks, acting as volume fluxes. Although elegant from a modelling perspective, the full multi-compartment Darcy system is computationally expensive to solve. We therefore enhance computational efficiency of this model by reducing the N-compartment system of Darcy equations to N pressure equations, and N subsequent projection problems to recover the Darcy velocity. The resulting 'reduced' Darcy formulation leads to a dramatic reduction in algebraic-system size and is therefore computationally cheaper to solve than the full multi-compartment Darcy system. A comparison of the reduced and the full formulation in terms of solution time and memory usage clearly highlights the superior performance of the reduced formulation. Moreover, the implementation of flux and, specifically, impermeable boundary conditions on arbitrarily curved boundaries such as epicardium and endocardium is straightforward in contrast to the full Darcy formulation. Finally, to demonstrate the applicability of our methodology to a personalized model and its solvability in clinically relevant time frames, we simulate perfusion in a subject-specific model of the left ventricle.


A miniaturized robotic platform for natural orifice transluminal endoscopic surgery: in vivo validation.
Natural orifice transluminal endoscopic surgery (NOTES) involves accessing the abdominal cavity via one of the body natural orifices for enabling minimally invasive surgical procedures. However, the constraints imposed by the access modality and the limited available technology make NOTES very challenging for surgeons. Tools redesign and introduction of novel surgical instruments are imperative in order to make NOTES operative in a real surgical scenario, reproducible and reliable. Robotic technology has major potential to overcome current limitations. The robotic platform described here consists of a magnetic anchoring frame equipped with dedicated docking/undocking mechanisms to house up to three modular robots for surgical interventions. The magnetic anchoring frame guarantees the required stability for surgical tasks execution, whilst dedicated modular robots provide the platform with adequate vision, stability and manipulation capabilities. Platform potentialities were demonstrated in a porcine model. Assessment was organized into two consecutive experimental steps, with a hybrid testing modality. First, platform deployment, anchoring and assembly through transoral-transgastric access were demonstrated in order to assess protocol feasibility and guarantee the safe achievement of the following experimental session. Second, transabdominal deployment, anchoring, assembly and robotic module actuation were carried out. This study has demonstrated the feasibility of inserting an endoluminal robotic platform composed of an anchoring frame and modular robotic units into a porcine model through a natural orifice. Once inserted into the peritoneal cavity, the platform provides proper visualization from multiple orientations. For the first time, a platform with interchangeable modules has been deployed and its components have been connected, demonstrating in vivo the feasibility of intra-abdominal assembly. Furthermore, increased dexterity employing different robotic units will enhance future system capabilities.


A review of the Nearctic Odontomachus (Hymenoptera: Formicidae: Ponerinae) with a treatment of the males.
The ant genus Odontomachus Latreille in the United States is reviewed. Six species are treated: O. brunneus (Patton), O. clarus Roger, O. desertorum Wheeler stat. nov., O. relictus Deyrup and Cover, O. ruginodis M.R. Smith, and O. haematodus (Linnaeus), a new record for North America. The spread of O. haematodus is documented, and its identity is clarified. The genus is diagnosed for species in the Nearctic region for all castes, and worker- and male-based keys are presented. The workers and males of all six species are described and figured, including the first male descriptions for O. haematodus and O. desertorum. This represents the first study of species-level variation in Odontomachus male genitalia, and one of the first of such studies of the Ponerinae for any biogeographic region. A discussion of the utility of the male sex for Odontomachus taxonomy is provided.


Degradation of p-toluenesulphonic acid via sidechain oxidation, desulphonation and meta ring cleavage in Pseudomonas (Comamonas) testosteroni T-2.
Pseudomonas (Comamonas) testosteroni T-2 completely converted p-toluenesulphonic acid (TS) or p-sulphobenzoic acid (PSB) to cell material, CO2 and sulphate, with growth yields of about 5 g protein (mol C)-1. PSB and sulphite were excreted as transient intermediates during growth in TS-salts medium. All reactions of a catabolic pathway involving sidechain oxidation and cleavage of the sulphonate moiety as sulphite were measurable in the soluble portion of cell extracts. Degradation of TS and PSB was inducible and apparently involved at least two regulons. TS was converted to p-sulphobenzyl alcohol in a reaction requiring NAD(P)H and 1 mol O2 (mol TS)-1. This alcohol was in an equilibrium (in the presence of NAD+) with p-sulphobenzaldehyde, which was converted to PSB in an NAD(P)+-dependent reaction. PSB was desulphonated to protocatechuic acid in a reaction requiring NAD(P)H and 1 mol O2 (mol PSB)-1. Experiments with 18 O2 confirmed involvement of a dioxygenase, because both atoms of this molecular oxygen were recovered in protocatechuate. Protocatechuate was converted to 2-hydroxy-4-carboxymuconate semialdehyde by a 4.5-dioxygenase.


Capacity for repeatable leukotriene generation after transient stimulation of mast cells and macrophages.
Leukotriene (LT) synthesis is initiated by the enzyme 5-lipoxygenase (5-LO). Prolonged cell stimulation causes the translocation of 5-LO to the nuclear envelope and the synthesis of LT, with subsequent inactivation and persistent membrane association of 5-LO. In this study, we examined whether persistent membrane association of 5-LO, as well as the inactivation of 5-LO, could be prevented by shortening the length of cell stimulation or by blocking LT synthesis. As expected, stimulation of rat basophilic leukaemia (RBL) cells, a mast cell model, or alveolar macrophages (AMs) with calcium ionophore for 15 min caused 5-LO translocation, LT generation and the inactivation and persistent membrane association of 5-LO. When RBL cells or AMs instead were stimulated for 0.5-5 min, translocation of 5-LO and synthesis of LT still occurred. However, after washing and resting, the 5-LO enzyme returned to its original intracellular distribution. Furthermore these cells showed a retained capacity for LT synthesis on subsequent re-stimulation. Similar results were obtained when cells were stimulated with either formyl peptide or zymosan, instead of ionophore. In contrast, blockade of LT synthesis during the initial stimulation, with the selective inhibitors zileuton or MK-886, did not inhibit 5-LO translocation, inactivation or persistent membrane association resulting from prolonged cell stimulation. We conclude that, in long-lived immune cells, 5-LO translocation is reversible when cell stimulation is short, but persistent after prolonged stimulation. In addition 5-LO remains active and LT synthetic capacity is retained after transient stimulation, whereas significant inactivation of 5-LO occurs after prolonged stimulation. Finally, results with LT synthesis inhibitors indicate that inactivation and persistent membrane association of 5-LO can result independently of 5-LO activation.


Ernest Henry Starling (1866-1927) on the formation and reabsorption of lymph.
Ernest Henry Starling laid the groundwork for our modern understanding of how the interstitial fluid, which he referred to as 'lymph', is regulated. Together with his colleague, William Bayliss, he provided the crucial insight into how fluid is driven out of the capillary to form interstitial fluid. That was to measure (estimate) the capillary pressure in different parts of the circulation and to relate changes in these pressures to altered lymph formation. In addressing how interstitial fluid re-enters the circulation, he was able to show that this occurs not only via the lymphatics, but also by re-entering the capillaries, mediated by the oncotic pressure of the plasma proteins. Starling's discoveries put to rest all notions that the processes of filtration and reabsorption of fluid are mediated by the 'vital activity' of cells. They could be explained entirely on the basis of physic-chemical forces. Based upon his insights from animal experiments, he was able to explain the genesis of edema (dropsy) in a number of disease states, including venous obstruction, cardiac disease and inflammatory conditions.


The Changing Face of Clinical Trials in Psoriatic Arthritis.
We will address current treatment and unmet needs in psoriatic arthritis (PsA), examine existing randomized controlled trials (RCTs), and consider options for new trial designs and challenges in their implementation. While therapeutic options for PsA have rapidly increased, there continues to be a need for clinical trials to test new therapies and establish optimal treatment strategies in order to improve the care for patients with PsA. In addition, more data is needed on how to select the best therapy for a given patient in clinical practice. Consideration of alternative outcome measures is also needed. Despite the rapid expansion in the number of therapy options available, there is still much to be learned about how to treat the individual patient with PsA.


SE33 locus as a reliable genetic marker for forensic DNA analysis systems
Background/aim: Genetic variation, an authentic tool of individual discrimination, is being used for forensic investigations worldwide. A missing result for even one out of 13-17 markers leads to an inconclusive report. Additional reliable markers are required to compensate such deficiencies. The SE33 locus has high genetic variability in different populations and is being used in forensic investigation systems in some countries. The purpose of the study was to assess the viability of use of the SE33 locus as a supportive marker for forensic DNA profiling. Materials and methods: Amplification of the SE33 locus was performed using the PowerPlex ES Monoplex System SE33 (Promega). After genotyping 204 Pakistani individuals, different genetic and forensic parameters for the SE33 locus were studied. Results: Genotyping of the SE33 locus revealed a total of 43 alleles including 3 novel alleles. Significant values of different forensic and genetic parameters including power of discrimination, power of exclusion, and polymorphism information content were observed. Conclusions: Addition of the SE33 locus in forensic DNA profiling may help to produce conclusive reports where results are inconclusive due to degraded evidence samples. The SE33 locus can confidently be used for Pakistani and neighboring populations having common ancestors from Iran to Central Asia, the Middle East, India and Turkey.


PCB elimination by yellow perch (Perca flavescens) during an annual temperature cycle.
The significance of temperature on aquatic species ecology and physiology is well recognized yet its effects on chemical bioaccumulation kinetics are less well understood under natural conditions. In this study, yellow perch were dosed with a polychlorinated biphenyl (PCB) mixture and allowed to depurate the chemicals over 1 year under an ambient temperature cycle characteristic of northern temperate latitudes. PCB elimination kinetics during the summer months at optimal water temperature for perch (23 degrees C) were similar to those observed in lab studies with other species reared at their optimal temperature. During the fall and winter seasons, however, elimination of only 11 PCB congeners of log K(ow) < or = 5.7 was observed and half-lives averaged > 1000 d for these PCBs. PCB elimination was again observed with the onset of spring temperatures but elimination rates averaged 2.6 times slower for readily metabolized congeners and 7.5 times slower for more persistent PCBs than observed during the summer. Bioenergetics modeling efforts predicted maximum values for respiration, fecal egestion, and growth rates during summer months but also predicted rapid declines in these chemical dilution processes during the fall and winter concurrent with changes in temperature. As temperature increased into the spring, bioenergetic rates were predicted to increase but only achieved approximately 85% of maximum rates predicted for summer peak temperatures. These results indicate that minimal chemical elimination occurs in perch when metabolic functioning falls to low maintenance levels during the fall and winter. These seasons encompass approximately 8 months of the year at northern temperate latitudes and therefore these patterns have significant consequences for understanding mechanisms of food-web biomagnification of hydrophobic organic chemicals in aquatic systems.


Opinion survey of analgesia for abdominal pain in Israeli emergency departments.
The long-standing and ongoing controversy regarding administration of analgesia to patients with acute abdominal pain prior to final diagnosis has not yet been resolved, despite considerable research. Consequently, wide variations in clinical practice exist. To determine the motives, attitudes and practices of emergency physicians, internists and surgeons in Israeli emergency departments regarding the administration of analgesia before diagnosis in patients with acute abdominal pain. Questionnaires were completed by 122 physicians in 21 EDs throughout Israel and the replies were analyzed. Most EDs did not have a clear policy on analgesia for undifferentiated abdominal pain, according to 65% of the responders. More internists (75%) than surgeons (54%) (P = 0.02) and more emergency physicians (81%) than all other physicians (60%) (P = 0.05) held this opinion. Most respondents (64%) supported administration of analgesia pre-diagnostically. Support for analgesia was significantly stronger among internists (75%) compared to surgeons (52%) (P = 0.03). Despite this wide support, most respondents (68%) indicated that analgesia was rarely or never given prediagnostically and, when it was, more surgeons (58%) than other physicians made that decision. Most internists (73%) and all surgeons reported that patients receive analgesia only after being examined by surgeons. Time allocated to the ED (part or full-time) significantly (P = 0.02) influenced decision-making, with 51% of part-time physicians vs. 25% of full-time opposing prompt administration of analgesia. Opinions on who should decide were divided according to medical specialty, with surgeons and internists almost opposed, as well as by physician age and percent of his/her time spent working in the ED. More surgeons than internists (P = 0.0005) reported that analgesia sometimes interfered with making a diagnosis. Most physicians (90%) stated that opiates impede diagnosis to some extent. However, 58% of them supported the administration of opiates, more or less frequently. Intramuscular diclofenac was the most preferred analgesic, followed by intravenous morphine and pethidine; individual preferences extended beyond the list of actually administered drugs. There is no consensus on the administration of analgesia for undiagnosed acute abdominal pain in EDs in Israel. Physicians' attitudes are influenced by training, experience, and percent of personal time allocated to work in the ED.


Inclusion Complexation of S-Nitrosoglutathione for Sustained Nitric Oxide Release from Catheter Surfaces: A Strategy to Prevent and Treat Device-Associated Infections.
S-Nitrosoglutathione (GSNO) is a nontoxic nitric oxide (NO)-donating compound that occurs naturally in the human body. The use of GSNO to deliver exogenous NO for therapeutic and protective applications is limited by the high lability of dissolved GSNO in aqueous formulations. In this paper, we report a host-guest chemistry-based strategy to modulate the GSNO reactivity and NO release kinetics for the design of anti-infective catheters and hydrogels. Cyclodextrins (CDs) are host molecules that are typically used to encapsulate hydrophobic guest molecules into their hydrophobic cavities. However, we found that CDs form inclusion complexes with GSNO, an extremely hydrophilic molecule with a solubility of over 1 M at physiological pH. More interestingly, the host-guest complexation reduces the decomposition reactivity of GSNO in the order of αCD > γCD > hydroxypropyl βCD. The lifetime of 0.1 M GSNO is increased to up to 15 days in the presence of CDs at 37 °C, which is more than twice the lifetime of free GSNO. Quantum chemistry calculations indicate that GSNO in αCD undergoes a conformational change that significantly reduces the S-NO bond distance and increases its stability. The calculated S-NO bond dissociation enthalpies of free and complexed GSNO well agree with the experimentally observed GSNO decomposition kinetics. The NO release from GSNO-CD solutions, compared to GSNO solutions, has suppressed initial bursts and extended durations, enhancing the safety and efficacy of NO-based therapies and device protections. In an example application as an anti-infective lock solution for intravascular catheters, the GSNO-αCD solution exhibits potent antibacterial activities for both planktonic and biofilm bacteria, both intraluminal and extraluminal environments, both prevention and treatment of infections, and against multiple bacterial strains, including a multidrug-resistant strain. In addition to solutions, the inclusion complexation also enables the preparation of GSNO hydrogels with enhanced stability and improved antibacterial efficacy. Since methods to suppress and control the GSNO decomposition rate are rare, this supramolecular strategy provides new opportunities for the formulation and application of this natural NO donor.


Glial cell line-derived neurotrophic factor (GDNF) is required for differentiation of pontine noradrenergic neurons and patterning of central respiratory output.
Genetic mutations affecting signaling by glial cell line-derived neurotrophic factor (GDNF) perturb development of breathing in mice and are associated with congenital central hypoventilation syndrome in humans. However, the role of GDNF in development of brainstem neurons that control breathing is largely unknown. The present study demonstrates that genetic loss of GDNF decreases the number of tyrosine hydroxylase (TH) neurons in the pontine A5 noradrenergic cell group, a major source of inhibitory input to the medullary respiratory pattern generator. This phenotype is associated with a significant increase in the frequency of central respiratory output recorded from the fetal medulla-spinal cord in vitro. In dissociate cultures of the A5 region from rat embryos, GDNF increases TH cell number and neurite growth without affecting total neuronal survival or proliferation of TH neurons. These effects of GDNF are inhibited by function blocking antibodies against endogenous brain-derived neurotrophic factor (BDNF), indicating that GDNF requires BDNF as a cofactor to stimulate differentiation of A5 neurons. Our findings demonstrate that GDNF is required for development of pontine noradrenergic neurons in vivo and indicate that defects in the A5 cell group may contribute to the effects of genetic disruption of GDNF signaling on respiratory control.


The influence of multiple trials and computer-mediated communication on collaborative and individual semantic recall.
Collaborative inhibition is a phenomenon where collaborating groups experience a decrement in recall when interacting with others. Despite this, collaboration has been found to improve subsequent individual recall. We explore these effects in semantic recall, which is seldom studied in collaborative retrieval. We also examine "parallel CMC", a synchronous form of computer-mediated communication that has previously been found to improve collaborative recall [Hinds, J. M., & Payne, S. J. (2016). Collaborative inhibition and semantic recall: Improving collaboration through computer-mediated communication. Applied Cognitive Psychology, 30(4), 554-565]. Sixty three triads completed a semantic recall task, which involved generating words beginning with "PO" or "HE" across three recall trials, in one of three retrieval conditions: Individual-Individual-Individual (III), Face-to-face-Face-to-Face-Individual (FFI) and Parallel-Parallel-Individual (PPI). Collaborative inhibition was present across both collaborative conditions. Individual recall in Recall 3 was higher when participants had previously collaborated in comparison to recalling three times individually. There was no difference between face-to-face and parallel CMC recall, however subsidiary analyses of instance repetitions and subjective organisation highlighted differences in group members' approaches to recall in terms of organisation and attention to others' contributions. We discuss the implications of these findings in relation to retrieval strategy disruption.


Micronutrient intake and breast cancer characteristics among postmenopausal women.
Few studies on micronutrients and postmenopausal breast cancer have examined the association with breast cancer characteristics. The aim of this study was to investigate the associations between vitamin C, vitamin E, folate and beta-carotene from diet and supplements and risk of postmenopausal breast cancer subtypes defined by histology (ductal/lobular), estrogen receptor (ER) and progesterone receptor (PGR) status. In a prospective cohort study of 26,224 postmenopausal women information on diet, supplements and lifestyle was collected through questionnaires. One thousand seventy-two cases were identified during follow-up. Incidence rate ratios of total breast cancers and breast cancer subtypes related to micronutrient intake were calculated using Cox proportional hazard analyses. This study found no association between overall breast cancer and any micronutrients, while some effects were shown when stratifying by breast cancer subtypes: dietary but not supplemental beta-carotene showed a protective effect against lobular breast cancer [incidence rate ratio (IRR): 0.72; 95% confidence interval (CI): 0.57-0.91]. Dietary vitamin E was associated with decreased risk of ER and PGR positive breast cancer (IRR: 0.50; 95% CI: 0.25-0.98) and dietary folate was associated with increased risk of ER and PGR positive breast cancer (IRR: 1.27; 95% CI: 1.03-1.95). This study found no effect of micronutrients on overall risk of postmenopausal breast cancer, but indicated possible effects of micronutrients in subgroups of breast cancer, with a potential beneficial effect of dietary beta-carotene in lobular breast cancer and dietary vitamin E in ER + PGR+ breast cancer and a potential harmful effect of dietary folate in ER+ PGR+ breast cancer.


Rapid qualitative urinary tract infection pathogen identification by SeptiFast real-time PCR.
Urinary tract infections (UTI) are frequent in outpatients. Fast pathogen identification is mandatory for shortening the time of discomfort and preventing serious complications. Urine culture needs up to 48 hours until pathogen identification. Consequently, the initial antibiotic regimen is empirical. To evaluate the feasibility of qualitative urine pathogen identification by a commercially available real-time PCR blood pathogen test (SeptiFast®) and to compare the results with dipslide and microbiological culture. Pilot study with prospectively collected urine samples. University hospital. 82 prospectively collected urine samples from 81 patients with suspected UTI were included. Dipslide urine culture was followed by microbiological pathogen identification in dipslide positive samples. In parallel, qualitative DNA based pathogen identification (SeptiFast®) was performed in all samples. 61 samples were SeptiFast® positive, whereas 67 samples were dipslide culture positive. The inter-methodological concordance of positive and negative findings in the gram+, gram- and fungi sector was 371/410 (90%), 477/492 (97%) and 238/246 (97%), respectively. Sensitivity and specificity of the SeptiFast® test for the detection of an infection was 0.82 and 0.60, respectively. SeptiFast® pathogen identifications were available at least 43 hours prior to culture results. The SeptiFast® platform identified bacterial DNA in urine specimens considerably faster compared to conventional culture. For UTI diagnosis sensitivity and specificity is limited by its present qualitative setup which does not allow pathogen quantification. Future quantitative assays may hold promise for PCR based UTI pathogen identification as a supplementation of conventional culture methods.


Subcortico-amygdala pathway processes innate and learned threats.
Behavioral flexibility and timely reactions to salient stimuli are essential for survival. The subcortical thalamic-basolateral amygdala (BLA) pathway serves as a shortcut for salient stimuli ensuring rapid processing. Here, we show that BLA neuronal and thalamic axonal activity in mice mirror the defensive behavior evoked by an innate visual threat as well as an auditory learned threat. Importantly, perturbing this pathway compromises defensive responses to both forms of threats, in that animals fail to switch from exploratory to defensive behavior. Despite the shared pathway between the two forms of threat processing, we observed noticeable differences. Blocking β-adrenergic receptors impairs the defensive response to the innate but not the learned threats. This reduced defensive response, surprisingly, is reflected in the suppression of the activity exclusively in the BLA as the thalamic input response remains intact. Our side-by-side examination highlights the similarities and differences between innate and learned threat-processing, thus providing new fundamental insights.


Near Infrared Light-Triggered Photocatalytic Decaging for Remote-Controlled Spatiotemporal Activation in Living Mice.
Spatiotemporal manipulation of biological processes in living animals using noninvasive, remote-controlled stimuli is a captivating but challenging endeavor. Herein, we present the development of a biocompatible photocatalytic technology termed CAT-NIR, which uses external near infrared light (NIR, 740 nm) to trigger decaging reactions in living mice. The Os(II) terpyridine complex was identified as an efficient NIR photocatalyst for promoting deboronative hydroxylation reactions via superoxide generation in the presence of NIR light, resulting in the deprotection of phenol groups and the release of bioactive molecules under living conditions. The validation of the CAT-NIR system was demonstrated through the NIR-triggered rescue of fluorophores, prodrugs as well as biomolecules ranging from amino acids, peptides to proteins. Furthermore, by combining genetic code expansion and computer-aided screening, CAT-NIR could regulate affibody binding to the cell surface receptor HER2, providing a selective cell tagging technology through external NIR light. In particular, the tissue-penetrating ability of NIR light allowed for facile prodrug activation in living mice, enabling noninvasive, remote-controlled rescue of drug molecules. Given its broad adaptability, this CAT-NIR system may open new opportunities for manipulating the functions of bioactive molecules in living animals using external NIR light with spatiotemporal resolution.


Absence of CTL responses to early viral antigens facilitates viral persistence.
CD8+ T cells are crucial for the control of intracellular pathogens such as viruses and some bacteria. Using lymphocytic choriomeningitis virus (LCMV) infection of mice--the prototypic arenavirus evolutionarily closely related to human Lassa fever and South American hemorrhagic fever viruses, we have shown previously that the kinetics of Ag presentation determine immunodominance of the LCMV-specific CTL response due to progressive exhaustion of LCMV nucleoprotein (NP)-specific CTL upon increasing viral load. In this study, we provide evidence that CTL against early LCMV NP-derived epitopes are more important in virus control than those against late glycoprotein-derived epitopes. We show that mice that are tolerant to all NP-derived T cell epitopes are severely compromised in their ability to control larger inocula of LCMV, supporting our hypothesis that CD8+ T cells specific for early viral Ags play a major role in acute virus control. Thus, the kinetics with which virus-derived T cell epitopes are presented has a strong impact on the efficacy of the antiviral immunity. This aspect should be taken into consideration for the development of vaccines.


Anemia in elderly hospitalized patients: prevalence and clinical impact.
Anemia is a common finding in elderly individuals. Several studies have shown a strong relationship between anemia, morbidity and mortality, suggesting anemia as a significant independent predictor of adverse outcome in elderly hospitalized patients. The pathophisiology of anemia in the elderly is not yet completely understood. Several mechanisms are involved. We investigated the prevalence of anemia in a cohort of 193 elderly patients admitted to the Internal Medicine Ward of Ca'Granda Policlinico Hospital along 6 months, and its relationship to comorbidities and to the length of hospitalization. Anemia was classified according to the WHO criteria. The majority of patients (48 %) had a mildmoderate, normocytic anemia; severe anemia was found in 8 out of 92 anemic patients. In a subgroup of patients erythropoietin was tested and resulted statistically higher if compared to non-anemic controls (p = 0.003). Considering the most common cause of anemia, nutritional deficiency, chronic renal disease and anemia of chronic disease were found respectively in 36, 15 and 25 % of cases. Unexplained anemia was diagnosed in 24 % of patients, according to the literature. Anemia was independently associated with increased length of hospital stay. Our study confirmed a high prevalence of anemia in elderly patients, and its association with a higher number of comorbidities and a longer stay. A correct clinical approach to anemia in elderly hospitalized patients is essential, considering its negative impact on patients' quality of life, and its social burden in term of healthcare needs and costs.


The promoter for C4-type mitochondrial aspartate aminotransferase does not direct bundle sheath-specific expression in transgenic rice plants.
For NAD-malic enzyme (NAD-ME)-type C4 photosynthesis, two types of aspartate aminotransferase (AAT) are involved. We examined the expression pattern of the Panicum miliaceum mitochondrial Aat gene (PmAat) and P. miliaceum cytosolic Aat gene (PcAat) in transgenic rice plants, which were specifically expressed in bundle sheath cells (BSCs) and mesophyll cells (MCs), respectively. Expression of a beta-glucuronidase (GUS) reporter gene under the control of the PcAat promoter was regulated in an organ-preferential and light-dependent manner in the transgenic rice plants. However, the PmAat promoter drove the GUS expression in all organs we tested without light dependency, and this non-preferential expression pattern was also observed in transgenic rice with introduction of the intact PmAat gene. The expression patterns of the rice counterpart Aat genes to PmAat or PcAat showed that the rice mitochondrial Aat (RmAat1) gene was expressed in all organs tested in a light-independent manner, while expression of the rice cytosolic Aat (RcAat1) gene showed an organ-preferential and light-dependent pattern. Taking these results together, we can generalize that the regulatory system of BSC-specific or light-dependent expression of mitochondrial Aat is not shared between P. miliaceum (C4) and rice (C3) and that the expression of the C4 genes introduced into rice mimics that of their counterpart genes in rice.


A Reconfigurable Sound Wave Decomposition Filterbank for Hearing Aids Based on Nonlinear Transformation.
Hearing impaired people have their own hearing loss characteristics and listening preferences. Therefore hearing aid system should become more natural, humanized and personalized, which requires the filterbank in hearing aids provides flexible sound wave decomposition schemes, so that patients are likely to use the most suitable scheme for their own hearing compensation strategy. In this paper, a reconfigurable sound wave decomposition filterbank is proposed. The prototype filter is first cosine modulated to generate uniform subbands. Then by non-linear transformation the uniform subbands are mapped to nonuniform subbands. By changing the control parameters, the nonlinear transformation changes which leads to different subbands allocations. It provides four different sound wave decomposition schemes without changing the structure of the filterbank. The performance of the proposed reconfigurable filterbank was compared with that of fixed filerbanks, fully customizable filterbanks and other existing reconfigurable filterbanks. It is shown that the proposed filterbank provides satisfactory matching performance as well as low complexity and delay, which make it suitable for real hearing aid applications.


A review of methodology and quantification in dental microwear analysis.
Dental microwear analysis is a method of inferring oral events (primarily food processing and aspects of masticatory biomechanics) from microscopic abrasion patterns retained on the enamel surfaces of teeth. Although some qualitative pattern differences may be easily distinguishable, most of the significant results produced thus far have derived from quantified studies of SEM images of occlusal enamel. It often goes unnoticed by readers of microwear reports who are not themselves specialists that microwear analysis is essentially a statistical method, not a visual one. In this review of current techniques and methods, several problems in current approaches are detailed. It is noted that feature definition can have significant effects on ultimate pattern differentiation. Sampling bias is also a major concern, as most microwear studies are carried out on samples which are very small. Compounding this are the effects of magnification level choices, and the effects of SEM instrumentation on feature visibility. Finally, the interpretation of pattern differences requires careful attention to comparisons of within-group and between-group variability.


HDAC2 regulates cell proliferation, cell cycle progression and cell apoptosis in esophageal squamous cell carcinoma EC9706 cells.
Increasing evidence has demonstrated that histone deacetylase 2 (HDAC2) participates in the regulation of a variety of biological processes in numerous tumors. However, the potential role of HDAC2 in the development and progression of esophageal squamous cell carcinoma (ESCC) remains elusive. Immunohistochemistry was utilized to detect the expression of HDAC2, Cell Counting Kit-8 was used to determine the cell proliferation, and flow cytometry was employed to investigate cell cycle and cell apoptosis. Finally, western blotting was employed to detect the protein expression of cyclin D1, p21, B cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax). The present study found that expression of HDAC2 protein in ESCC tissues was significantly increased compared with atypical hyperplasia tissues and normal esophageal mucosa (P<0.001). The expression of HDAC2 was not associated with the age or gender of patients (P>0.05), but was closely associated with the histological grade, invasion depth, tumor-node-metastasis stage and lymph node metastasis, respectively (all P<0.001). HDAC2 small interfering RNA effectively downregulated the expression of HDAC2 protein in ESCC EC9706 cells. Downregulation of HDAC2 expression evidently inhibited cell proliferation, arrested cell cycle at the G0/G1 phase and induced cell apoptosis in ESCC EC9706 cells, coupled with increased expression of p21 and Bax proteins and decreased expression of cyclin D1 and Bcl-2 proteins. Overall, the present findings suggest that HDAC2 may play an important role in the development and progression of ESCC and be considered as a novel molecular target for the treatment of ESCC.


Endobronchial tuberculosis--a report of 5 cases.
Endobronchial tuberculosis is not as well-known to internists as tuberculosis involving the lung parenchyma. Five cases with this condition are reported to illustrate the varied clinical manifestations. The presenting features of the 5 cases were lobar or lung collapse, unresolved pneumonia, dyspnoea and stridor. Bronchostenosis developed in 2 patients many years following chemotherapy, while stenosis of the trachea developed in one patient during chemotherapy. In another patient, the tuberculous granulation tissue simulated a lung cancer at bronchoscopy. Diagnosis can be difficult as endobronchial tuberculosis can occur in the absence of chest X-ray abnormality and sputum smear may also be negative for acid fast bacilli (AFB). Therefore, bronchoscopy should be done when the condition is suspected in a patient who has unexplained cough, wheezing, dyspnoea or haemoptysis. The modalities of treatment for fibrostenosis of a large airway include surgical resection followed by anastomosis, balloon dilatation, laser photoresection or a combination of both procedures.


Modulation of long-term depression by dopamine in the mesolimbic system.
Long-lasting adaptations in the mesolimbic dopamine (DA) system in response to drugs of abuse likely mediate many of the behavioral changes that underlie addiction. Recent work suggests that long-term changes in synaptic strength at excitatory synapses in the two major components of this system, the nucleus accumbens (NAc) and ventral tegmental area, may be particularly important for the development of drug-induced sensitization, a process that may contribute to addiction, as well as for normal response-reinforcement learning. Using whole-cell patch-clamp recording techniques from in vitro slice preparations, we have examined the existence and basic mechanisms of long-term depression (LTD) at excitatory synapses on both GABAergic medium spiny neurons in the NAc and dopaminergic neurons in the midbrain. We find that both sets of synapses express LTD but that their basic triggering mechanisms differ. Furthermore, DA blocks the induction of LTD in the midbrain via activation of D2-like receptors but has minimal effects on LTD in the NAc. The existence of LTD in mesolimbic structures and its modulation by DA represent mechanisms that may contribute to the modifications of neural circuitry that mediate reward-related learning as well as the development of addiction.


AIDS Clinical Research in Spain-Large HIV Population, Geniality of Doctors, and Missing Opportunities.
The first cases of AIDS in Spain were reported in 1982. Since then over 85,000 persons with AIDS have been cumulated, with 60,000 deaths. Current estimates for people living with HIV are of 145,000, of whom 20% are unaware of it. This explains the still high rate of late HIV presenters. Although the HIV epidemic in Spain was originally driven mostly by injection drug users, since the year 2000 men having sex with men (MSM) account for most new incident HIV cases. Currently, MSM represent over 80% of new yearly HIV diagnoses. In the 80s, a subset of young doctors and nurses working at Internal Medicine hospital wards became deeply engaged in attending HIV-infected persons. Before the introduction of antiretrovirals in the earlier 1990s, diagnosis and treatment of opportunistic infections was their major task. A new wave of infectious diseases specialists was born. Following the wide introduction of triple combination therapy in the late 1990s, drug side effects and antiretroviral resistance led to built a core of highly devoted HIV specialists across the country. Since then, HIV medicine has improved and currently is largely conducted by multidisciplinary teams of health care providers working at hospital-based outclinics, where HIV-positive persons are generally seen every six months. Antiretroviral therapy is currently prescribed to roughly 75,000 persons, almost all attended at clinics belonging to the government health public system. Overall, the impact of HIV/AIDS publications by Spanish teams is the third most important in Europe. HIV research in Spain has classically been funded mostly by national and European public agencies along with pharma companies. Chronologically, some of the major contributions of Spanish HIV research are being in the field of tuberculosis, toxoplasmosis, leishmaniasis, HIV variants including HIV-2, drug resistance, pharmacology, antiretroviral drug-related toxicities, coinfection with viral hepatitis, design and participation in clinical trials with antiretrovirals, immunopathogenesis, ageing, and vaccine development.


Use of plasmon coupling to reveal the dynamics of DNA bending and cleavage by single EcoRV restriction enzymes.
Pairs of Au nanoparticles have recently been proposed as "plasmon rulers" based on the dependence of their light scattering on the interparticle distance. Preliminary work has suggested that plasmon rulers can be used to measure and monitor dynamic distance changes over the 1- to 100-nm length scale in biology. Here, we substantiate that plasmon rulers can be used to measure dynamical biophysical processes by applying the ruler to a system that has been investigated extensively by using ensemble kinetic measurements: the cleavage of DNA by the restriction enzyme EcoRV. Temporal resolutions of up to 240 Hz were obtained, and the end-to-end extension of up to 1,000 individual dsDNA enzyme substrates could be simultaneously monitored for hours. The kinetic parameters extracted from our single-molecule cleavage trajectories agree well with values obtained in bulk through other methods and confirm well known features of the cleavage process, such as DNA bending before cleavage. Previously unreported dynamical information is revealed as well, for instance, the degree of softening of the DNA just before cleavage. The unlimited lifetime, high temporal resolution, and high signal/noise ratio make the plasmon ruler a unique tool for studying macromolecular assemblies and conformational changes at the single-molecule level.


A prospective analysis of laboratory tests and imaging studies to detect hepatic lesions.
A prospective study of the ability of laboratory tests and liver imaging tests to detect hepatic metastases was performed. Eighty patients at risk for hepatic metastases but without clinical evidence of disease were tested with 13 laboratory tests and three liver imaging tests. No single laboratory test had greater than 65% accuracy in the detection of hepatic lesions. No combination of the laboratory tests increased this accuracy. If the laboratory tests were used with one of the liver imaging tests, the accuracy was improved in some combinations to 76%. The CEA assay when analyzed in patients with colorectal primaries had an accuracy of 79%. The results show that the laboratory tests alone are not sufficiently accurate to detect liver metastases. Additional accuracy can be obtained by the combined use of a single liver imaging test and selected laboratory tests. Use of all the liver imaging tests and laboratory tests lowers the accuracy and increases the expense and thus is unnecessary.


Tissue-specific target analysis of disease-associated microRNAs in human signaling pathways.
MicroRNAs are a large class of post-transcriptional regulators that bind to the 3' untranslated region of messenger RNAs. They play a critical role in many cellular processes and have been linked to the control of signal transduction pathways. Recent studies indicate that microRNAs can function as tumor suppressors or even as oncogenes when aberrantly expressed. For more general insights of disease-associated microRNAs, we analyzed their impact on human signaling pathways from two perspectives. On a global scale, we found a core set of signaling pathways with enriched tissue-specific microRNA targets across diseases. The function of these pathways reflects the affinity of microRNAs to regulate cellular processes associated with apoptosis, proliferation or development. Comparing cancer and non-cancer related microRNAs, we found no significant differences between both groups. To unveil the interaction and regulation of microRNAs on signaling pathways locally, we analyzed the cellular location and process type of disease-associated microRNA targets and proteins. While disease-associated proteins are highly enriched in extracellular components of the pathway, microRNA targets are preferentially located in the nucleus. Moreover, targets of disease-associated microRNAs preferentially exhibit an inhibitory effect within the pathways in contrast to disease proteins. Our analysis provides systematic insights into the interaction of disease-associated microRNAs and signaling pathways and uncovers differences in cellular locations and process types of microRNA targets and disease-associated proteins.


Evaluation of an enzyme-linked immunosorbent assay based on crude Leishmania histone proteins for serodiagnosis of human infantile visceral leishmaniasis.
Human visceral leishmaniasis (VL) is routinely diagnosed by detecting IgG that specifically binds to Leishmania antigens. The enzyme-linked immunosorbent assay (ELISA) remains a widely used method. However, the biggest challenge remains the choice of antigen with the highest specificity and sensitivity. This study is aimed at assessing the diagnostic performances of crude Leishmania histone (CLH) protein-based ELISAs in Mediterranean VL patients. The CLH proteins were biochemically purified from promastigote nuclear extracts. Their reactivities were analyzed by Western blotting (WB) using rabbit polyclonal antibodies against Leishmania recombinant histones and sera from VL patients, respectively. Then, the diagnostic potential of CLH proteins was validated by the CLH-based ELISA using 42 infantile VL patients' sera and 70 control subjects. The CLH-based ELISA performance was compared to that of the soluble Leishmania antigen (SLA)- and the recombinant K39 (rK39)-based ELISAs. Analysis of the WB profile with the use of polyclonal antibodies confirmed the histone origin of low molecular mass proteins (12 to 16 kDa). All VL samples tested presented antibodies reacting against different antigen fractions; however, recognition patterns were different depending on the reactivity of each serum. CLH-based ELISA showed an excellent ability to discriminate between VL cases and healthy controls (97.6% sensitivity and 100% specificity). It had a diagnostic performance similar to that of rK39-based ELISA (97.6% sensitivity and 97.1% specificity, P = 0.5) and a better serodiagnosis accuracy than the SLA-based ELISA (85.7% sensitivity and 90% specificity, P < 0.05). Therefore, crude Leishmania histone extract could be a valuable antigen for clinical use.


Environmentally controlled dimorphic cycle in a fission yeast.
The fission yeast Schizosaccharomyces pombe shows bipolar growth and is a convenient model for studying cell polarity and polar growth. This paper shows that the related Schiz. japonicus var. japonicus can switch to unipolar growth and can exist in both yeast and mycelial phases. On solid media, the yeast phase is unstable and prone to switch to the mycelial form, which shows unipolar growth by tip elongation. The hyphae can colonize the body of the substrate (true mycelium) or just its surface (pseudo-mycelium). The yeast-to-mycelium transition and the growth of the mycelium are regulated by a nutritional gradient and are associated with extensive vacuolation. The mycelium can convert into arthroconidia or return to the yeast phase in response to environmental changes. These environmentally controlled morphological transitions make Schiz. japonicus var. japonicus an attractive model for the investigation of cell polarity and morphogenesis.


Structural dissimilarity from self drives neoepitope escape from immune tolerance.
T-cell recognition of peptides incorporating nonsynonymous mutations, or neoepitopes, is a cornerstone of tumor immunity and forms the basis of new immunotherapy approaches including personalized cancer vaccines. Yet as they are derived from self-peptides, the means through which immunogenic neoepitopes overcome immune self-tolerance are often unclear. Here we show that a point mutation in a non-major histocompatibility complex anchor position induces structural and dynamic changes in an immunologically active ovarian cancer neoepitope. The changes pre-organize the peptide into a conformation optimal for recognition by a neoepitope-specific T-cell receptor, allowing the receptor to bind the neoepitope with high affinity and deliver potent T-cell signals. Our results emphasize the importance of structural and physical changes relative to self in neoepitope immunogenicity. Considered broadly, these findings can help explain some of the difficulties in identifying immunogenic neoepitopes from sequence alone and provide guidance for developing novel, neoepitope-based personalized therapies.


Associations Between Variations in TPH1 , TPH2 and SLC6A4 Genes and Postpartum Depression: A Study in the Jordanian Population.
In this study, we investigated the association between tryptophan hydroxylase-1 (TPH1 ) (218A>C), tryptophan hydroxylase-2 ( TPH2 ) (1463G>A) and serotonin carrier family 6, member 4 (SLC6A4) [long (L) vs. short (S)] gene polymorphisms with post-partum depression (PPD) in women from Jordan. A total of 370 postpartum (130 depressed and 240 non depressed) women volunteered for the study. Genotyping was carried out using restriction fragment length polymorphism (RFLP) for TPH1 , amplification refractory mutation system (ARMS) for TPH2 and polymerase chain reaction (PCR) for SLC6A4 S and L. The Edinburgh postnatal depression scale was used to screen postpartum women. Both S and L alleles of SLC6A4 are common in Jordanian women (about 51.0 and 49.0%, respectively), while allele TPH1 -218C is more common (64.0%) than allele A (37.0%). Regarding TPH2 , allele A is absent from the examined women. None of the examined polymorphisms were found to be associated with PPD ( p >0.05). However, depression history, pregnancy problems and economic status were found to be significantly associated with PPD ( p <0.05). The results suggest that TPH1 , TPH2 and SLC6A4 S and L polymorphisms do not seem to be important in Jordan for predisposing to PPD.


The amount of sonic hedgehog in multiple sclerosis white matter is decreased and cleavage to the signaling peptide is deficient.
We have demonstrated that sonic hedgehog (Shh), vital for oligodendrocyte development, is present in both gray and white matter of normal human brain. Both the 45 kDa precursor protein and the 20 kDa N-terminal sonic hedgehog signaling portion (ShhN) were demonstrated by immunoblot and a partial purification has been achieved. In gray matter from brains of multiple sclerosis (MS) victims, the total amount of Shh was less than normals and the signaling 20 kDa protein was greatly reduced. In white matter homogenates, prepared from MS victims, only the 45 kDa precursor protein was found. None of the 20 kDa signaling protein was detected, suggesting that the 45 kDa signaling protein was not cleaved in the autocatalytic reaction carried out by the C-terminal portion. The 45 kDa protein and a small amount of the 20 kDa ShhN was detected in isolated MS myelin by Western blot, demonstrating some cleavage was possible. The cleavage of the 45 kDa protein was demonstrated in normal myelin in vitro, but not in myelin prepared from MS brain.


Efficacy of low-dose methotrexate treatment in birdshot chorioretinopathy.
To ascertain the effect of treatment with methotrexate (MTX) on the visual prognosis of birdshot chorioretinopathy (BSCR). Retrospective case series of 76 consecutive patients with HLA-A29-positive BSCR, of whom 46 were followed for at least 5 years and 18 for longer than 10 years. A review of the medical records of 76 patients with BSCR. Treatment regimens were subdivided into the following groups: 1) No systemic immunomodulatory treatment; 2) Treatment with systemic corticosteroids; and 3) Treatments which comprised MTX. First, we calculated eye-years for the different therapeutic regimens and second, we subdivided the patients according to their initial treatment regimen and assessed visual outcomes. Mean visual acuity increased over time in the MTX-treated patients; remained unchanged in patients on systemic corticosteroids and decreased in the patients without systemic treatment (yearly change in LogMar -0.020, -0.034 and 0.028 with P = 0.034, P = 0.71 and P = 0.006 respectively). In the group treated initially with MTX, VA gradually increased in contrast to the remaining groups of patients (P = 0.003). In this series, treatment comprising MTX showed better visual outcomes than the untreated patients and corticosteroid-based treatment regimens.


Amyloidoma of the gasserian ganglion.
We report an unusual case of an amyloidoma of the gasserian ganglion associated with trigeminal neuralgia. MR imaging showed a mass in Meckel's cave, which was isointense with surrounding tissue on T1-weighted images and hypointense on T2-weighted images. After contrast administration, the mass enhanced homogeneously, and thin cuts through the region showed involvement in the expected location of the gasserian ganglion and, more laterally and inferiorly, in the proximal part of V1. A review of the literature revealed that only one similar case has been reported since the advent of modern neuroimaging. Amyloidoma, although rare, may be considered as a rare differential diagnosis of a mass in this region.


Characterisation and potential diagnostic value of circulating matrix Gla protein (MGP) species.
Matrix γ-carboxyglutamate (Gla) protein (MGP) is an important local inhibitor of vascular calcification, which can undergo two post-translational modifications: vitamin K-dependent γ-glutamate carboxylation and serine phosphorylation. While carboxylation is thought to have effects upon binding of calcium-ions, phosphorylation is supposed to affect the cellular release of MGP. Since both modifications can be exerted incompletely, various MGP species can be detected in the circulation. MGP levels were measured with two commercially available competitive and two novel sandwich assays in healthy controls, in patients with rheumatic disease, aortic valve disease, and end-stage renal disease, as well as in volunteers after vitamin K supplementation (VKS) and treatment with vitamin K antagonists (VKA). Major differences were found between the MGP assays, including significantly different behaviour with regard to vascular disease and the response to VKA and VKS. The dual-antibody assay measuring non-phosphorylated, non-carboxylated MGP (dp-ucMGP) was particularly sensitive for these changes and would be suited to assess the vascular vitamin K status. We conclude that the different assays for particular circulating MGP species allows the assessment of various aspects of the MGP system.


Targeting the Sonic Hedgehog Signaling Pathway: Review of Smoothened and GLI Inhibitors.
The sonic hedgehog (Shh) signaling pathway is a major regulator of cell differentiation, cell proliferation, and tissue polarity. Aberrant activation of the Shh pathway has been shown in a variety of human cancers, including, basal cell carcinoma, malignant gliomas, medulloblastoma, leukemias, and cancers of the breast, lung, pancreas, and prostate. Tumorigenesis, tumor progression and therapeutic response have all been shown to be impacted by the Shh signaling pathway. Downstream effectors of the Shh pathway include smoothened (SMO) and glioma-associated oncogene homolog (GLI) family of zinc finger transcription factors. Both are regarded as important targets for cancer therapeutics. While most efforts have been devoted towards pharmacologically targeting SMO, developing GLI-targeted approach has its merit because of the fact that GLI proteins can be activated by both Shh ligand-dependent and -independent mechanisms. To date, two SMO inhibitors (LDE225/Sonidegib and GDC-0449/Vismodegib) have received FDA approval for treating basal cell carcinoma while many clinical trials are being conducted to evaluate the efficacy of this exciting class of targeted therapy in a variety of cancers. In this review, we provide an overview of the biology of the Shh pathway and then detail the current landscape of the Shh-SMO-GLI pathway inhibitors including those in preclinical studies and clinical trials.


Association of Oral Candida albicans with Severe Early Childhood Caries - A Pilot Study.
In early childhood, children are more susceptible to opportunistic microbial colonization in the oral cavity due to immature immune system and not fully established micro flora. The current literature proposes a probable role of Candida albicans, a fungus in the etiopathogenesis of dental caries. This study was conducted to compare the Candida albicans count in children with severe early childhood caries and caries free children. A cross-sectional study was conducted in 40 randomly selected healthy children between 12 to 71 months of age, who were divided into two groups based on the caries experience as Severe Early Childhood Caries (SECC) (dmfs ≥4) and caries free (dmfs = 0). The caries experiences (dmfs index) of the 40 children were recorded using visible light and diagnostic instruments. A 2ml sample of unstimulated whole saliva collected from the children was transported to the microbiology laboratory in universal containers and evaluated for Candida albicans count using the selective media. The data was statistically analyzed using SPSS software 17.0. Candida albicans was found in both the SECC group and caries free group. Median Candida albicans of the SECC group was numerically greater than the caries free group and this difference was highly statistically significant (p=0.012). In this present cross-sectional study, we found a 100% prevalence of Candida albicans in the saliva of the study children. There was a highly significant increase in Candida albicans count in SECC children compared to the caries free children.


Probing of the reactive center loop region of alpha-1-antitrypsin by mutagenesis predicts new type-2 dysfunctional variants.
Lung disease in alpha-1-antitrypsin deficiency (AATD) mainly results from insufficient control of the serine proteases neutrophil elastase (NE) and proteinase-3 due to reduced plasma levels of alpha-1-antitrypsin (AAT) variants. Mutations in the specificity-determining reactive center loop (RCL) of AAT would be predicted to minimally affect protein folding and secretion by hepatocytes but can impair anti-protease activity or alter the target protease. These properly secreted but dysfunctional 'type-2' variants would not be identified by common diagnostic protocols that are predicated on a reduction in circulating AAT. This has potential clinical relevance: in addition to the dysfunctional Pittsburgh and Iners variants reported previously, several uncharacterized RCL variants are present in genome variation databases. To prospectively evaluate the impact of RCL variations on secretion and anti-protease activity, here we performed a systematic screening of amino acid substitutions occurring at the AAT-NE interface. Twenty-three AAT variants that can result from single nucleotide polymorphisms in this region, including 11 present in sequence variation databases, were expressed in a mammalian cell model. All demonstrated unaltered protein folding and secretion. However, when their ability to form stable complexes with NE was evaluated by western blot, enzymatic assays, and a novel ELISA developed to quantify AAT-NE complexes, substrate-like and NE-binding deficient dysfunctional variants were identified. This emphasizes the ability of the RCL to accommodate inactivating substitutions without impacting the integrity of the native molecule and demonstrates that this class of molecule violates a generally accepted paradigm that equates circulating levels with functional protection of lung tissue.


SARS-CoV-2 Spike Protein Induces Hemagglutination: Implications for COVID-19 Morbidities and Therapeutics and for Vaccine Adverse Effects.
Experimental findings for SARS-CoV-2 related to the glycan biochemistry of coronaviruses indicate that attachments from spike protein to glycoconjugates on the surfaces of red blood cells (RBCs), other blood cells and endothelial cells are key to the infectivity and morbidity of COVID-19. To provide further insight into these glycan attachments and their potential clinical relevance, the classic hemagglutination (HA) assay was applied using spike protein from the Wuhan, Alpha, Delta and Omicron B.1.1.529 lineages of SARS-CoV-2 mixed with human RBCs. The electrostatic potential of the central region of spike protein from these four lineages was studied through molecular modeling simulations. Inhibition of spike protein-induced HA was tested using the macrocyclic lactone ivermectin (IVM), which is indicated to bind strongly to SARS-CoV-2 spike protein glycan sites. The results of these experiments were, first, that spike protein from these four lineages of SARS-CoV-2 induced HA. Omicron induced HA at a significantly lower threshold concentration of spike protein than the three prior lineages and was much more electropositive on its central spike protein region. IVM blocked HA when added to RBCs prior to spike protein and reversed HA when added afterward. These results validate and extend prior findings on the role of glycan bindings of viral spike protein in COVID-19. They furthermore suggest therapeutic options using competitive glycan-binding agents such as IVM and may help elucidate rare serious adverse effects (AEs) associated with COVID-19 mRNA vaccines, which use spike protein as the generated antigen.


The association of plasma lactate with incident cardiovascular outcomes: the ARIC Study.
We examined the association of plasma lactate at rest, a marker of oxidative capacity, with incident cardiovascular outcomes in 10,006 participants in the Atherosclerosis Risk in Communities (ARIC) Study visit 4 (1996-1998). We used Cox proportional-hazards models to estimate hazard ratios of incident coronary heart disease, stroke, heart failure, and all-cause mortality by quartiles of plasma lactate (Q1, ≤5.3 mg/dL; Q2, 5.4-6.6; Q3, 6.7-8.6; and Q4 ≥8.7). During a median follow-up time of 10.7 years, there were 1,105 coronary heart disease cases, 379 stroke cases, 820 heart failure cases, and 1,408 deaths. A significant graded relation between lactate level and cardiovascular events was observed in the demographically adjusted model (all P for trend < 0.001). After further adjustment for traditional and other potential confounders, the association remained significant for heart failure (Q4 vs. Q1: hazard ratio (HR) = 1.35, 95% confidence interval (CI): 1.07, 1.71) and all-cause mortality (HR = 1.27, 95% CI: 1.07, 1.51) (P for trend < 0.02 for these outcomes) but not for coronary heart disease (HR = 1.02, 95% CI: 0.84, 1.24) and stroke (HR = 1.26, 95% CI: 0.91, 1.75). The results for heart failure were robust across multiple subgroups, after further adjustment for N-terminal pro-B-type natriuretic peptide and after exclusion of participants with incident heart failure within 3 years. The independent associations of plasma lactate with heart failure and all-cause mortality suggest an important role for low resting oxidative capacity.


MHC-I Genotype Restricts the Oncogenic Mutational Landscape.
MHC-I molecules expose the intracellular protein content on the cell surface, allowing T cells to detect foreign or mutated peptides. The combination of six MHC-I alleles each individual carries defines the sub-peptidome that can be effectively presented. We applied this concept to human cancer, hypothesizing that oncogenic mutations could arise in gaps in personal MHC-I presentation. To validate this hypothesis, we developed and applied a residue-centric patient presentation score to 9,176 cancer patients across 1,018 recurrent oncogenic mutations. We found that patient MHC-I genotype-based scores could predict which mutations were more likely to emerge in their tumor. Accordingly, poor presentation of a mutation across patients was correlated with higher frequency among tumors. These results support that MHC-I genotype-restricted immunoediting during tumor formation shapes the landscape of oncogenic mutations observed in clinically diagnosed tumors and paves the way for predicting personal cancer susceptibilities from knowledge of MHC-I genotype.


Reprogramming the Local Lymph Node Microenvironment Promotes Tolerance that Is Systemic and Antigen Specific.
Many experimental therapies for autoimmune diseases, such as multiple sclerosis (MS), aim to bias T cells toward tolerogenic phenotypes without broad suppression. However, the link between local signal integration in lymph nodes (LNs) and the specificity of systemic tolerance is not well understood. We used intra-LN injection of polymer particles to study tolerance as a function of signals in the LN microenvironment. In a mouse MS model, intra-LN introduction of encapsulated myelin self-antigen and a regulatory signal (rapamycin) permanently reversed paralysis after one treatment during peak disease. Therapeutic effects were myelin specific, required antigen encapsulation, and were less potent without rapamycin. This efficacy was accompanied by local LN reorganization, reduced inflammation, systemic expansion of regulatory T cells, and reduced T cell infiltration to the CNS. Our findings suggest that local control over signaling in distinct LNs can promote cell types and functions that drive tolerance that is systemic but antigen specific.


18-Fluorodeoxyglucose Uptake by Positron Emission Tomography in Extraocular Muscles of Patients with and without Graves' Ophthalmology.
Objective. To compare 18-fluorodeoxyglucose (FDG) uptake by positron emission tomography (PET) in extraocular muscles (EOMs) of patients with Graves' ophthalmopathy (GO) versus patients without GO. Design. Prospective, observational, comparative, and cross-sectional study. Participants. Thirty-two eyes of patients with GO and seventy eyes of patients without GO. Methods. We prospectively included patients older than 18 years of age with and without GO. FDG-PET imaging study was performed; standardized unit value (SUVmax) was quantified in EOMs. Standard deviation and significant statistical difference (P < 0.05) were calculated. Results. Thirty-two eyes of sixteen patients of the GO group were included, with a mean age of 44.31 (20-71) years. Seventy eyes of thirty-five patients of the group without GO were included, with a mean age of 49.20 (24-77) years. EOMs average uptake of the groups with and without GO were 3.38 ± 1.31 and 1.89 ± 0.51 SUVmax (P < 0.05), respectively. Conclusion. FDG uptake was significantly increased in EOMs of patients with GO. PET gives valuable information and may be a helpful tool in detecting, localizing, and quantifying GO inflammation. Further research is needed to define the role of PET in detecting, grading, and following up GO in order to optimize treatment in the inflammatory stage.


Evaluation of the Potential of Collagen from Codfish Skin as a Biomaterial for Biomedical Applications.
Collagen is one of the most widely used biomaterials, not only due its biocompatibility, biodegradability and weak antigenic potential, but also due to its role in the structure and function of tissues. Searching for alternative collagen sources, the aim of this study was to extract collagen from the skin of codfish, previously obtained as a by-product of fish industrial plants, and characterize it regarding its use as a biomaterial for biomedical application, according to American Society for Testing and Materials (ASTM) Guidelines. Collagen type I with a high degree of purity was obtained through acid-extraction, as confirmed by colorimetric assays, SDS-PAGE and amino acid composition. Thermal analysis revealed a denaturing temperature around 16 °C. Moreover, collagen showed a concentration-dependent effect in metabolism and on cell adhesion of lung fibroblast MRC-5 cells. In conclusion, this study shows that collagen can be obtained from marine-origin sources, while preserving its bioactivity, supporting its use in biomedical applications.


The effect of curcumin (active substance of turmeric) on the acetic acid-induced visceral nociception in rats.
In the present study, the effect of chronic oral administration of curcumin in the presence or absence of morphine and noloxone was investigated on the visceral nociception induced by acetic acid in rats. Intraperitoneal injection of acetic acid (1 mL, 2%) produced contractions in the abdominal musculature (writhes). The latency time to the beginning of the first writhe was measured and the total number of writhes in the 1 h after acetic acid injection was counted. The latency time to the beginning of the first writhe was significantly (p < 0.05) increased and the number of writhes was significantly (p < 0.05) decreased by curcumin (20 and 40 mg kg(-1) body weight). The same results were obtained after subcutaneous injection of morphine (1 mg kg(-1) b.wt.). Naloxone at the dose of 1 mg kg(-1) body weight had no effect on pain intensity. Curcumin significantly (p < 0.05) enhanced the effect of morphine on the visceral pain responses, however did not reverse the effect of naloxone. Present data suggest that in the acetic acid-induced visceral nociception of rats, curcumin may produce an antinociceptive effect and the endogenous analgesic opioid system is involved in the curcumin-induced antinociception.


Molecular Dynamics Study on Structure, Vibrational Properties, and Transport Coefficients of Liquid Alumina.
The structure, vibrational density of states, and transport coefficients of liquid alumina were studied using molecular dynamics simulations. At the temperature of 2500 K, 3000 K, 3500 K, and 4000 K, systems with three different densities were constructed, respectively, including the configurations with densities of 2.81 g/cm3 and 3.17 g/cm3, and the relaxed ones with nearly zero pressure at each temperature. With the changes in temperature or density, the transformations on the structural, vibrational and transport properties were discussed. The Born-Mayer-Huggins type of atomic interactions was used, with newly optimized parameters. The analysis of the interatomic correlations indicated that the short-range order of liquid alumina was mainly constructed by AlO4 tetrahedra, also a certain number of AlO3 and AlO5 was present. Meanwhile, the structural transitions on the elemental units occurred as either the temperature or density increased. Two primary frequency bands were observed in each vibrational density of states spectrum, with the higher frequency bands produced by the O atom vibrations, and the lower frequency ones generated by the Al atom vibrations. Self-diffusion coefficients were estimated using the linear behavior of the mean-squared displacement at long time, while by using the Green-Kubo relation during equilibrium molecular dynamics simulations, thermal conductivities and viscosities were calculated. Significantly, the viscosity at 2500 K with a density of 2.81 g/cm3 was equal to 25.23 mPa s, which was very close to the experimental finding.


Plant phenology shifts under climate warming: a systematic review of recent scientific literature.
Climate warming-driven temporal shifts in phenology are widely recognised as the foremost footprint of global environmental change. In this regard, concerted research efforts are being made worldwide to monitor and assess the plant phenological responses to climate warming across species, ecosystems and seasons. Here, we present a global synthesis of the recent scientific literature to assess the progress made in this area of research. To achieve this, we conducted a systematic review by following PRISMA protocol, which involved rigorous screening of 9476 studies on the topic and finally selected 215 studies for data extraction. The results revealed that woody species, natural ecosystems and plant phenological responses in spring season have been predominantly studied, with the herbaceous species, agricultural ecosystems and other seasons grossly understudied. Majority of the studies reported phenological advancement (i.e., preponement) in spring, followed by also advancement in summer but delay in autumn. Methodology-wise, nearly two -third of the studies have employed direct observational approach, followed by herbarium-based and experimental approaches, with the latter covering least temporal depth. We found a steady increase in research on the topic over the last decade with a sharp increase since 2014. The global country-wide scientific output map highlights the huge geographical gaps in this area of research, particularly in the biodiversity-rich tropical regions of the developing world. Based on the findings of this global synthesis, we identify the current knowledge gaps and suggest future directions for this emerging area of research in an increasingly warming world.


How Mixed Relay Teams in Swimming Should Be Organized for International Championship Success.
The primary goal of the present research was to determine the order of swimmers on a mixed relay team that would ensure the best performance in the Fédération Internationale de Natation (FINA) World Championships held in Kazan (Russia, 2015), Budapest (Hungary, 2017), and Gwangju (South Korea, 2019). The data were obtained from database websites for the 4 × 100 m freestyle and 4 × 100 m medley official results, including 660 records from 188 entries of finals and 472 preliminary events. The results showed that the fastest swimmers (according to their best season times) were located primarily in the first or second positions of the freestyle relay. The most successful gender strategy for the 4 × 100 m freestyle (57 out of 82 observations) and for the 4 × 100 m medley (29 out of 83) relays was the order male-male-female-female, although no statistical differences were found (p = 0.79) for the medley relays. In the 4 × 100 m freestyle, the second (p = 0.002; β = 1.62) and third (p =0.003; β = 1.41) relay legs had a statistical effect on the total relay time, whereas in the 4 × 100 m medley, all four relay legs had a statistical effect (p < 0.001) on the final performance, the weight of the four strokes being different in heats with respect to the final round. Also, a later position of the first female swimmer or the consecutive position of two female swimmers in the team order significantly affected the relay performance in specific events. Mixed relay events appeared to present specific strategies in comparison to traditional male- or female-only relay lineups.


Different biodegradation potential and the impacted soil functions of epoxiconazole in two soils.
Epoxiconazole is an effective pesticide to control Fusarium head blight (FHB), and the application will increase. To investigate the ecotoxicity of epoxiconazole to soil microbiome, we carried out an indoor experiment in which soils from two main regions of wheat production in China (Nanjing and Anyang) were treated with epoxiconazole (0, 0.0625, 0.625, or 6.25 mg kg-1) and incubated for 90 days. Under epoxiconazole stress, for bacteria and fungi, the abundance was increased and the diversity and community were impacted. In Anyang soil, the half-life of epoxiconazole was short with more increased species (linear discriminant analysis effect size biomarkers) and more increased xenobiotics biodegradation pathways in epoxiconazole treatments. The increased species mostly due to high abundance in initial state and more positive connections of the species. Co-occurrences revealed that epoxiconazole tightened bacterial connection, and increased positive correlations in Anyang soil. The N transformation was influenced with increased nifH and amoA; and the contents of NH4+-N and NO3--N were also increased. The functions of C, S, and manganese metabolisms were also impacted by epoxiconazole. This work expands our understanding about epoxiconazole degradation and help us to properly assess the risk of epoxiconazole in soil.


Age-associated changes in protein oxidation and proteasome activities in rat brain: modulation by antioxidants.
The free radical theory of ageing postulates that age-associated neurodegeneration is caused by an imbalance between pro-oxidants and antioxidants resulting in oxidative stress. The current study showed regional variation in brain susceptibility to age-associated oxidative stress as shown by increased lipofuscin deposition and protein carbonyl levels in male rats of age 15-16 months compared to control ones (3-5 months). The hippocampus is the area most vulnerable to change compared to the cortex and cerebellum. However, proteasomal enzyme activity was not affected by age in any of the brain regions studied. Treatment with melatonin or coenzyme Q10 for 4 weeks reduced the lipofuscin content of the hippocampus and carbonyl level. However, both melatonin and coenzyme Q10 treatments inhibited beta-glutamyl peptide hydrolase activity. This suggests that these molecules can alter proteasome function independently of their antioxidant actions.