Suppression of autoimmune disease and of massive lymphadenopathy in MRL/Mp-lpr/lpr mice lacking tyrosine kinase Fyn (p59fyn).
MRL/Mp-lpr/lpr (MRL/lpr) mice suffer from a generalized autoimmune disease that includes autoantibody production and glomerulonephritis and develop massive lymphadenopathy characterized by an expanded population of CD4- CD8- B220+ T cells that is derived from autoreactive T cells in the periphery. Some of us previously reported that these atypical T cells overexpressed a gene for tyrosine kinase p59fyn (Fyn). To define the role of Fyn in the renal disease and lymphadenopathy in MRL/lpr mice, we have generated Fyn-deficient MRL/lpr mice whose fyn gene is replaced by the gene for beta-galactosidase. Fyn-deficient MRL/lpr mice developed markedly limited disease and lived more than twice as long as the conventional MRL/lpr mice. In the mutant mice, the production of IgG3 anti-DNA autoantibody was significantly (p < 0.005%) reduced, and glomerular deposits of IgG3 and C3 were remarkably diminished. Ag receptor-mediated proliferative responses of Fyn-deficient splenic T cells were markedly impaired. The mutant mice showed delayed accumulation of the atypical CD4- CD8- B220+ T cells that exhibited a significantly lower activity of ZAP-70 compared with those in the conventional MRL/lpr mice. These data demonstrated that Fyn is involved as a positive regulator in the disease of MRL/lpr mice. Fyn provides a signal for both the expansion of autoreactive T cells and the production of IgG3 anti-DNA autoantibody by B cells. Thus, manipulation of Fyn may improve systemic autoimmune disease in humans.


Cholesterol Emboli Co-Existing with Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis in a 76-Year-Old Woman.
Anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis injures small vessels and causes severe systemic organ injury. Main target antigens of ANCA are myeloperoxidase and proteinase 3. ANCA strongly associates with the development and progression of the vasculitis. Its manifestations include rapidly progressive glomerulonephritis, interstitial pneumonitis, alveolar hemorrhage, purpura, and neurological disorder. Most patients with ANCA-associated vasculitis in Japan are elderly and have atherosclerotic risk factors. Cholesterol emboli are systemic vascular inflammation triggered by cholesterol crystals. Cholesterol emboli cause kidney dysfunction and ischemia of the intestines, brain, heart, skin, and peripheral nerves. Diabetes mellitus, hypertension, hyperlipidemia, and history of cardiovascular diseases are risk factors of the development of cholesterol emboli. We report a case of ANCA-associated vasculitis coexisting with cholesterol emboli. A 76-year-old woman was diagnosed with ANCA-associated interstitial pneumonitis. She rapidly developed progressive glomerulonephritis, purpura, and peripheral sensory nerve disorder. A kidney biopsy revealed that renal dysfunction was caused by vasculitis of the interlobular arteries and cholesterol emboli. A skin biopsy revealed that purpura was caused by cholesterol emboli. Glucocorticoid and statin therapies were administered. Thereafter, the renal function and other symptoms improved and stabilized. The representative symptoms of ANCA-associated vasculitis and cholesterol emboli are closely similar, and it is difficult to distinguish between these diseases when they coexist. Because the background characteristics of patients with ANCA-associated vasculitis and risk factors of cholesterol emboli overlap, at the time of diagnosing ANCA-associated vasculitis, clinicians should consider the possibility of cholesterol emboli coexistence.


Phosphoenolpyruvate carboxylase identified as a key enzyme in erythrocytic Plasmodium falciparum carbon metabolism.
Phospoenolpyruvate carboxylase (PEPC) is absent from humans but encoded in the Plasmodium falciparum genome, suggesting that PEPC has a parasite-specific function. To investigate its importance in P. falciparum, we generated a pepc null mutant (D10(Δpepc) ), which was only achievable when malate, a reduction product of oxaloacetate, was added to the growth medium. D10(Δpepc) had a severe growth defect in vitro, which was partially reversed by addition of malate or fumarate, suggesting that pepc may be essential in vivo. Targeted metabolomics using (13)C-U-D-glucose and (13)C-bicarbonate showed that the conversion of glycolytically-derived PEP into malate, fumarate, aspartate and citrate was abolished in D10(Δpepc) and that pentose phosphate pathway metabolites and glycerol 3-phosphate were present at increased levels. In contrast, metabolism of the carbon skeleton of (13)C,(15)N-U-glutamine was similar in both parasite lines, although the flux was lower in D10(Δpepc); it also confirmed the operation of a complete forward TCA cycle in the wild type parasite. Overall, these data confirm the CO2 fixing activity of PEPC and suggest that it provides metabolites essential for TCA cycle anaplerosis and the maintenance of cytosolic and mitochondrial redox balance. Moreover, these findings imply that PEPC may be an exploitable target for future drug discovery.


An efficient and recyclable enzyme catalytic system constructed through the synergy between biomimetic mineralization and polyamine-salt aggregate assembly.
In the present study, an efficient and easily recyclable enzyme catalytic system based on nanoparticle-assembled microcapsules (NAMCs) is constructed by using both biomimetic mineralization and polyamine-salt aggregate assembly. Specifically, enzyme-encapsulated silica nanoparticles (NPs) are prepared by polyethyleneimine (PEI, a typical cationic polymer)-induced silicification in the presence of enzymes. The as-acquired NPs are then spontaneously adsorbed and assembled onto the surfaces of poly (allylamine hydrochloride) (PAH)-citrate microaggregates, which are synthesized through citrate-induced PAH aggregation. After continuous stirring, the internal parts of the PAH-citrate microaggregates are disassembled, and the NAMCs-based enzyme catalytic system is finally acquired. During the continuous stirring process, the PAH moieties on the surfaces of PAH-citrate microaggregates play two crucial roles: (1) adsorbing NPs onto the surfaces of PAH-citrate microaggregates by electrostatic interactions; and (2) catalyzing the subsequent condensation of the Si-OH groups on the surfaces of two adjacent NPs to form Si-O-Si networks, thus generating an intact capsule wall. In this process, the citrate moieties in the internal parts of PAH-citrate microaggregates are released into the bulk aqueous solution, and this release causes the disassembly of PAH-citrate microaggregates, thus generating the capsule lumen. The resultant enzyme catalytic system exhibits high activity and stability, and particularly, easy recyclability for converting formaldehyde into methanol using NADH as the cofactor. More specifically, this system displays an NADH conversion of ∼92.7%, and can be nearly completely recycled using low-speed centrifugation (<3000 rpm), which ensures that the NADH conversion is maintained almost unaltered after reusing the system 10 times. This study will be useful for the facile construction of diverse catalytic systems with high efficiency and excellent recyclability.


The climatic rhythm and blooms of cyanobacteria in a tropical reservoir in São Paulo, Brazil.
The present study sought to develop a methodology to analyse water quality based on the concepts and methods of climate and climatology. Accordingly, we attempted to relate hydro- and limnometeorological techniques and methodologies to a rhythmic analysis technique developed within the context of the Brazilian geographical climatology. Our goal was to assess and analyse cyanobacterial blooms, the main index of water quality for the reservoirs of the "Alto Tietê" Basin and, consequently, the Metropolitan Region of São Paulo, an area of high environmental complexity due to its high degree of development and high population density. The meteorological data used were collected by the Institute of Astronomy, Geophysics and Atmospheric Sciences at the University of São Paulo meteorological station, and the limnological data were collected through the Hydrological Monitoring System implemented by SABESP in the Billings and Guarapiranga Reservoirs and the laboratory of the same entity. The rhythmic and integrated analysis showed that the process of cyanobacterial blooms is dependent on a combination of meteorological factors as temperature and wind intensity that may disrupt the stability of the reservoir, providing the conditions necessary for the development of cyanobacteria during the stabilisation process. The pace of the Atlantic Polar Front Entrance during the winter in São Paulo is a limiting factor for the growth of cyanobacteria because of their high frequency, thus maintaining the balance of the reservoir throughout this period. The weather types those could cause a instability in the water column were: Cold Front entrance (66.67%), conflict between masses (22.22%) and the Tropical Instability Line (11.11%). The possibility for prevention and forecasting periods advise when these reservoirs should not be used, mainly with regard to recreational activities.


Identification and evaluation of outcome measurement instruments in pyoderma gangrenosum: a systematic review.
Pyoderma gangrenosum (PG) is a rare autoinflammatory skin condition that causes tissue destruction and subsequent painful ulcers. To date, there are no core domains or instruments for assessing PG severity in clinical trials, and current treatment paradigms rely on outcome measurements that have not been well characterized in the literature. To perform two systematic reviews that (i) identify the outcome measurement instruments used in PG clinical trials and their corresponding domains and (ii) identify any associated validation studies and evaluate their measurement properties and methodological quality. We systematically searched the MEDLINE and Embase databases for PG outcome measurement instruments. We also systematically searched for PG instrument validation studies. We evaluated the measurement properties and methodological quality of validation studies using the 2018 COSMIN Risk of Bias checklist. In total, seven clinical trials were included. These studies utilized a total of 20 different instruments, including 11 physician-reported instruments, eight patient-reported instruments and one composite instrument. Among these, 85% of the instruments lacked any validation data. Of the remaining three validated instruments (speed of healing, physician global assessment and resolution of inflammation), methodological quality was not available for half of the COSMIN categories. We identified 17 non-validated outcome measurement instruments used in PG clinical trials. We conclude that PG validation studies are required for existing instruments, and new instruments need to be developed to inform the consensus process for the development of a core outcome set for PG. What is already known about this topic? Pyoderma gangrenosum (PG) is a rare autoinflammatory skin condition that has been characterized by multiple outcome measurement instruments in clinical trials. However, there is no consensus on the most validated and appropriate outcome measurement instruments. What does this study add? This study identifies and evaluates 20 unique outcome measurement instruments for PG in the literature. Of these 20, 17 lack any instrument validation data, highlighting the need for future studies. What are the clinical implications of this work? Despite the current use of several outcome measurement instruments, future studies should explore the validation surrounding these instruments, as no instruments can currently be recommended.


Blocking human fear memory with the matrix metalloproteinase inhibitor doxycycline.
Learning to predict threat is a fundamental ability of many biological organisms, and a laboratory model for anxiety disorders. Interfering with such memories in humans would be of high clinical relevance. On the basis of studies in cell cultures and slice preparations, it is hypothesised that synaptic remodelling required for threat learning involves the extracellular enzyme matrix metalloproteinase (MMP) 9. However, in vivo evidence for this proposal is lacking. Here we investigate human Pavlovian fear conditioning under the blood-brain barrier crossing MMP inhibitor doxycyline in a pre-registered, randomised, double-blind, placebo-controlled trial. We find that recall of threat memory, measured with fear-potentiated startle 7 days after acquisition, is attenuated by ~60% in individuals who were under doxycycline during acquisition. This threat memory impairment is also reflected in increased behavioural surprise signals to the conditioned stimulus during subsequent re-learning, and already late during initial acquisition. Our findings support an emerging view that extracellular signalling pathways are crucially required for threat memory formation. Furthermore, they suggest novel pharmacological methods for primary prevention and treatment of posttraumatic stress disorder.


[Removal of lipoprotein complexes of cholesterol from plasma using silochromes].
Industrial silicates, silochromes, proved to be effective sorbents of cholesterol lipoprotein complexes in blood in vitro. Silochrome SH-1.5 with a diameter of pores of 180-200 nm and a capacity of 12-14 mg of cholesterol per 1 g of the sorbent exhibited the best efficiency in isolation of the complexes. At the same time, maximal unspecific binding of blood plasma proteins was observed in the systems with silochrome SH-3 with a diameter of pores of 60 nm. By use of silochrome SH-1.5 for plasmosorption the content of cholesterol was decreased from 3,600 mg/l down to 2,150 mg/l, i.e. by 40%, in blood plasma of a rabbit with experimental hypercholesterolemia.


Regulation of pathways degrading aromatic substrates in Pseudomonas putida. Enzymic response to binary mixtures of substrates.
1. Induction constants (K(ind)) and repression constants (K(rep)), which are a measure of the affinity of the inducers or repressors for the induction systems, were measured for mandelate, benzoate and p-hydroxybenzoate in Pseudomonas putida. 2. From these results, the enzymic response of the organism to media containing pairs of these substrates was predicted. Nitrogen-limited chemostats, operated at high growth rates, were used to investigate these predictions in cells grown first on one aromatic substrate with the second added later. 3. In general, the values of K(ind) and K(rep) predicted quite accurately the response to substrate mixtures. Thus, in the presence of mandelate and either benzoate or p-hydroxybenzoate, the enzymes of mandelate metabolism were repressed almost completely, and the bacteria were fully induced for the alternative substrate (benzoate or p-hydroxybenzoate), which was preferentially utilized for growth. When benzoate and p-hydroxybenzoate were the two substrates in the mixture, the enzymes for metabolism of the latter were strongly repressed and growth took place mainly on benzoate. 4. The enzymic response to mixed substrates did not result in the metabolism of the better growth substrate, but in the substrate requiring the synthesis of fewer enzymes. Thus benzoate is used in preference to mandelate although the latter supports a faster growth rate. It is nevertheless considered that, with our present knowledge of the natural habitat of the organism, it is impossible to decide whether protein economy or growth rate was the factor determining the evolution of this control system.


Importance of N- and C-terminal regions of gastrin-Gly for preferential binding to high and low affinity gastrin-Gly receptors.
G17-Gly has been shown to stimulate the growth of DLD-1 human colon cancer cells in a biphasic manner via high and low affinity receptors. In the current study, the existence of heterogeneous receptor populations for G17-Gly on the HT-29 human colon cancer cell line was investigated. The effect of either N- or C-terminal peptide truncation on receptor binding and cell growth stimulation was also explored. [Leu15]G17-Gly bound to both high (nM) and low (microM) affinity sites on HT-29 cells. The peptide stimulated cell growth in a dose-dependent and biphasic manner with maximal stimulation at 10(-9) M peptide concentration, suggesting that, as in the case of DLD-1 cells, it is the high affinity receptor which is responsible for the growth-promoting effects. In contrast, G17(1-12) stimulated the growth of HT-29 cells in a sigmoidal fashion with an EC50 of 4.6x10(-9) M. Sequential N-terminal truncation of [Leu15]G17-Gly results in decreased binding to the high affinity G17-Gly receptor on DLD-1 cells. [Leu15]G17(11-17)Gly bound to the low affinity G17-Gly receptor with an affinity similar to that of the full sequence peptide but was unable to displace the radioligand from high affinity sites. G17(1-6)-NH2 was unable to displace [3H]G17-Gly from either site. These results suggest that the important residues for binding to the low affinity receptor are in the C-terminal region of the peptide while those required for interaction with the high affinity receptor lie further towards the N-terminus.


Liver metastasis of endometrial stromal sarcoma.
Resection of liver metastases from gynaecological tumours is uncommon. Endometrial stromal sarcomas (ESS) are low incidence gynecological tumours which can originate in previous sites of endometriosis or following metaplasia of the pelvic peritoneal wall, and which are exceptionally associated with liver metastasis. We present a 68-year-old woman with a ESS and metachronic liver metastasis treated by liver resection. There is very little literature on clinical management about liver metastasis from ESS, but extrapolating the data obtained with liver metastasis from sarcomas and uterine tumours, we should recommend resection as this is considered a resectable extrauterine metastasis. In cases with more sites of extrauterine disease, liver resection should also be performed if the other sites are resectable.


Identifying Patient Access Barriers for Tumor Necrosis Factor Alpha Inhibitor Treatments in Rheumatoid Arthritis in Five Central Eastern European Countries.
Although there is a significant utilization gap of biologic medicines in the EU, many studies estimate equity in patient access to biopharmaceuticals only based on their availability on the national list of reimbursed medicines. Hidden access barriers may facilitate financial sustainability of pharmaceuticals in less affluent EU countries; however, they have rarely been documented in scientific publications. Our objective was to explore these access barriers for tumor necrosis factor (TNF) alpha inhibitors in rheumatoid arthritis (RA) in five Central and Eastern European countries. A detailed interview guide was developed based on multi-stakeholder workshops and a targeted literature review. In each participant country 3-3-3-3 interviews with payers, rheumatologists, patients/patient representatives, and industry representatives were conducted. Responses were aggregated at a country level and validated by primary investigators in each country. Limited number of RA centers and consequently significant travelling time and cost for patients in distant geographical areas, uneven budget allocation among centers, limited capacity of nurses, narrowed patient population in national financial protocols compared to international clinical guidelines in initiating or continuing biologics, high administrative burden in prescribing biologics and limited health literacy of patients were the most relevant barriers to timely patient access in at least three participant countries. Assessing only the availability of TNF alpha inhibitors on the national list of reimbursed medicines provides limited information about real-world patient access to these medicines. Revealing hidden access barriers may contribute to initiate policy actions which could reduce inequity in patient access.


Environmental impacts of synthetic fishing nets from manufacturing to disposal: A case study of Türkiye in life cycle perspective.
Fishing nets have a limited lifespan, and they are considered waste when worn or damaged beyond repair. If the end-of-life fishing nets (EOLFNs) are not adequately managed, they can be wasted in landfills or discarded into the sea inappropriately. Thus, encouraging the recycling of fishing nets is an effective way to reduce waste and promote a circular economy. This study examines the environmental impact of all processes of synthetic fishing nets used in various fishing operations (gillnet, trammel net and cast net) from a life-cycle perspective. The functional unit is a 1 ton fishing net. The disposal method represents two different scenarios, incineration and recycling. It has been observed that the disposal of EOLFNs by recycling rather than incineration offers significant environmental gains in all impact categories. The findings offer a promising solution to reduce the environmental impacts of EOLFNs by appropriately intervening in the effective recycling of fishing nets.


Hip arthrodesis: a procedure for the new millennium?
Advanced hip arthrosis in the young patient represents a difficult problem. Using currently available techniques, total hip arthroplasty (THA) in this patient population likely would result in multiple revision surgeries, usually at the expense of bone in the proximal femur and acetabulum. For noninflammatory, monoarticular hip arthritis, arthrodesis remains an excellent reconstruction option. Arthrodesis can provide pain relief and return to function in the majority of patients indicated for the procedure. Current techniques provide acceptable fusion rates. If possible, techniques that protect the abductor mechanism and limit deformity of the pelvis should be used. Patient education is an important part of the treatment process. The position of the limb is important to early satisfaction and durability of the fusion. Long-term difficulties after hip fusion are related to pain in contiguous joints. Conversion arthroplasty, if necessary, can result in favorable outcomes. Familiarity with the techniques and outcomes of hip arthrodesis are important for the hip surgeon to understand.


Clinical application of growth factors and cytokines in wound healing.
Wound healing is a complex and dynamic biological process that involves the coordinated efforts of multiple cell types and is executed and regulated by numerous growth factors and cytokines. There has been a drive in the past two decades to study the therapeutic effects of various growth factors in the clinical management of nonhealing wounds (e.g., pressure ulcers, chronic venous ulcers, diabetic foot ulcers). For this review, we conducted an online search of Medline/PubMed and critically analyzed the literature regarding the role of growth factors and cytokines in the management of these wounds. We focused on currently approved therapies, emerging therapies, and future research possibilities. In this review, we discuss four growth factors and cytokines currently being used on and off label for the healing of wounds. These include granulocyte-macrophage colony-stimulating factor, platelet-derived growth factor, vascular endothelial growth factor, and basic fibroblast growth factor. While the clinical results of using growth factors and cytokines are encouraging, many studies involved a small sample size and are disparate in measured endpoints. Therefore, further research is required to provide definitive evidence of efficacy.


A Four-Phase Model of Transdisciplinary Team-Based Research: Goals, Team Processes, and Strategies.
The complexity of social and public health challenges has led to burgeoning interest and investments in cross-disciplinary team-based research, and particularly in transdisciplinary (TD) team-based research. TD research aims to integrate and ultimately extend beyond discipline-specific concepts, approaches, and methods to accelerate innovations and progress toward solving complex real-world problems. While TD research offers the promise of novel, wide-reaching and important discoveries, it also introduces unique challenges. In particular, today's investigators are generally trained in unidisciplinary approaches, and may have little training in, or exposure to, the scientific skills and team processes necessary to collaborate successfully in teams of colleagues from widely disparate disciplines and fields. Yet these skills are essential to maximize the efficiency and effectiveness of TD team-based research. In the current article we propose a model of TD team-based research that includes four relatively distinct phases: development, conceptualization, implementation, and translation. Drawing on the science of team science (SciTS) field, as well as the findings from previous research on group dynamics and organizational behavior, we identify key scientific goals and team processes that occur in each phase and across multiple phases. We then provide real-world exemplars for each phase that highlight strategies for successfully meeting the goals and engaging in the team processes that are hallmarks of that phase. We conclude by discussing the relevance of the model for TD team-based research initiatives, funding to support these initiatives, and future empirical research that aims to better understand the processes and outcomes of TD team-based research.


Enhanced Excited-State Proton Transfer via a Mixed Methanol-Water Molecular Bridge of 1-Naphthol-3,6-disulfonate in Methanol-Water Mixtures.
Steady-state and time-resolved fluorescence techniques were used to study the excited-state proton transfer (ESPT) from an irreversible photoacid, 1-naphthol-3,6-disulfonate (1NP36DS), to methanol-water mixtures. We found that at χwater = 0.3 the ESPT rate constant is higher by a factor of 10 that in neat methanol. TD-DFT calculations show that a mixed molecular bridge of two methanol molecules and one water molecule enables the ESPT from the 1-OH to the 3-sulfonate. The RO-(S1) state is stable by -2.5 kcal/mol in comparison to the ROH(S1) state. We compare the ESPT rate constants of a reversible photoacid, 8-hydroxy-1,3,6-pyrenetrisulfonate (HPTS), in the same methanol-water mixtures. At χwater ≈ 0.3 the ESPT rate constant of HPTS increased by only 15%. We explain the large difference of the ESPT rate of 1NP36DS by the formation of a water bridge or a mixed methanol-water bridge from 1-OH to one of the sulfonates and the absence of such a bridge in HPTS. The water or mixed methanol-water bridge of 1NP36DS enhances the ESPT rate in methanol-water mixtures of low water mole ratio.


Evaluation of the Reinforcing Strength of Phendimetrazine Using a Progressive-Ratio Schedule of Reinforcement in Rhesus Monkeys.
Stimulant abuse is a persistent public health problem with no Food and Drug Administration-approved pharmacotherapy. Although monoamine-releasing drugs such as d-amphetamine can decrease cocaine self-administration in human and animal laboratory studies, their potential for abuse limits clinical utility. "Abuse-deterrent" formulations of monoamine releasers, such as prodrugs, hold greater clinical promise if their abuse potential is, as theorized, lower than that of cocaine. In these studies, we determined the reinforcing strength of phendimetrazine (PDM), a prodrug for the amphetamine-like monoamine releaser phenmetrazine; both drugs have been shown to decrease cocaine self-administration in laboratory animals. To date, no study has directly compared PDM (Schedule III) with cocaine (Schedule II) under progressive-ratio (PR) schedules of reinforcement, which are better suited than fixed-ratio schedules to directly compare reinforcing strength of drugs. Dose-response curves for cocaine (saline, 0.001-0.3 mg/kg per injection) and PDM (0.1-1.0 mg/kg per injection) were generated in six cocaine-experienced male rhesus monkeys during 4-hour sessions with a 20-minute limited hold (LH). Under these conditions, the maximum number of injections was not significantly different between cocaine and PDM. The reinforcing strength of doses situated on the peaks of the cocaine and PDM dose-effect curves were redetermined with a 60-minute LH. The mean number of injections increased for both drugs, but not for saline. Cocaine presentations resulted in significantly higher peak injections than PDM with a 60-minute LH, which is consistent with the lower scheduling of PDM. These results support PDM as Schedule III and highlight the importance of schedule parameters when comparing reinforcing strength of drugs using a PR schedule of reinforcement. SIGNIFICANCE STATEMENT: One strategy for reducing cocaine use is to identify a treatment that substitutes for cocaine but has lower abuse potential. In a rhesus monkey model of drug abuse, this study compared the reinforcing strength of cocaine and phendimetrazine, a drug that has been shown to decrease cocaine use in some studies.


Observational and ecological studies of dietary advanced glycation end products in national diets and Alzheimer's disease incidence and prevalence.
Considerable evidence indicates that diet is an important risk-modifying factor for Alzheimer's disease (AD). Evidence is also mounting that dietary advanced glycation end products (AGEs) are important risk factors for AD. This study strives to determine whether estimated dietary AGEs estimated from national diets and epidemiological studies are associated with increased AD incidence. We estimated values of dietary AGEs using values in a published paper. We estimated intake of dietary AGEs from the Washington Heights-Inwood Community Aging Project (WHICAP) 1992 and 1999 cohort studies, which investigated how the Mediterranean diet (MeDi) affected AD incidence. Further, AD prevalence data came from three ecological studies and included data from 11 countries for 1977-1993, seven developing countries for 1995-2005, and Japan for 1985-2008. The analysis used dietary AGE values from 20 years before the AD prevalence data. Meat was always the food with the largest amount of AGEs. Other foods with significant AGEs included fish, cheese, vegetables, and vegetable oil. High MeDi adherence results in lower meat and dairy intake, which possess high AGE content. By using two different models to extrapolate dietary AGE intake in the WHICAP 1992 and 1999 cohort studies, we showed that reduced dietary AGE significantly correlates with reduced AD incidence. For the ecological studies, estimates of dietary AGEs in the national diets corresponded well with AD prevalence data even though the cooking methods were not well known. Dietary AGEs appear to be important risk factors for AD.


Comparison of drug survival on adalimumab, etanercept, golimumab and infliximab in patients with axial spondyloarthritis.
To compare drug survival in patients with axial spondyloarthritis treated with different TNF inhibitors in standard dosage. Patients fulfilling the Assessment in SpondyloArthritis international Society classification criteria for axial spondyloarthritis in the Swiss Clinical Quality Management cohort were included in this study if a first TNF inhibitor on standard dosage was started after recruitment and if a baseline visit was available. Drug maintenance up to drug discontinuation or dose escalation was compared between TNF inhibitors with multiple adjusted Cox proportional hazards models and multiple imputation for missing baseline covariate data. A total of 966 patients were included (adalimumab 344, etanercept 237, golimumab 214, infliximab 171). Patients on certolizumab (n = 18) were excluded. Patients starting golimumab had lower disease activity as well as better physical function and quality of life in comparison to patients starting another drug. A higher proportion of patients starting infliximab had a history of extra-articular manifestations. Drug dosage was more often escalated during follow-up in patients treated with infliximab than with subcutaneously administered agents. However, no significant differences in time up to drug discontinuation or dose escalation were observed in multiple adjusted analyses if treatment was initiated after 2009, when all 4 TNF inhibitors were available: hazard ratio for infliximab versus etanercept 1.16 (95% confidence interval 0.80; 1.67), p = 0.44, for golimumab versus etanercept 0.80 (0.58; 1.10), p = 0.17 and for adalimumab versus etanercept 0.93 (0.69; 1.26), p = 0.66. In axial spondyloarthritis, drug survival with standard doses of different TNF inhibitors is comparable.


Effects of organoselenium compounds on induction of mouse forestomach tumors by benzo(a)pyrene.
The selenium analogues of three known inhibitors of chemical carcinogenesis were synthesized and the compounds were tested for their ability to inhibit the induction of forestomach tumors in mice by benzo(a)pyrene. Groups of female CD-1 mice were given NIH-07 diet, or NIH-07 diet to which one of the following test compounds had been added: p-methoxyphenol (30 mumol/g diet and 3.3 mumol/g diet); p-methoxybenzeneselenol (3.3 mumol/g diet); benzylthiocyanate (0.045 mumol/g diet); benzylselenocyanate (0.045 mumol/g diet); phenothiazine (3.8 mumol/g diet); and phenoselenazine (3.8 mumol/g diet). The test compounds were administered for 1 week prior to treatment with benzo(a)pyrene, during the 4 weeks of benzo(a)pyrene treatment, and for 1 week after benzo(a)pyrene treatment. Twelve weeks later the mice were sacrificed and forestomach tumors were counted and confirmed histologically as papillomas. p-Methoxyphenol was the most effective inhibitor and was the only one which significantly reduced both the percentage of tumor-bearing animals and the number of forestomach tumors per animal. At the 3.3-mumol/g diet, p-methoxyphenol reduced the number of tumors per animal from 3.3 to 0.8 (P less than 0.0003). p-Methoxybenzeneselenol reduced the number of tumors per animal from 3.3 to 2.0 (P less than 0.05). Benzylthiocyanate showed no significant inhibitory effect, but benzylselenocyanate reduced the number of tumors per animal from 3.3 to 1.7 (P less than 0.01). Phenothiazine significantly enhanced the number of tumors per animal from 3.3 to 6.5 (P less than 0.004). Phenoselenazine had no effect on tumor induction. The results of this study indicate that two synthetic organoselenium compounds, p-methoxybenzeneselenol and benzylselenocyanate, are effective inhibitors of mouse forestomach tumorigenesis induced by benzo(a)pyrene.


Dose-dependent pharmacokinetics of amphotericin B lipid complex in rabbits.
Amphotericin B lipid complex (ABLC) was recently approved by the Food and Drug Administration for treatment of patients with invasive fungal infections who are intolerant of or refractory to conventional amphotericin B therapy. Little is known, however, about the pharmacokinetics of this new antifungal compound. We therefore investigated the pharmacokinetics of ABLC in comparison with those of conventional desoxycholate amphotericin B (DAmB) in rabbits. The pharmacokinetics of DAmB in a rabbit model were similar to those previously reported in humans. The pharmacokinetics of ABLC differed substantially from those of DAmB. Plasma amphotericin B levels following ABLC administration were 10 times lower than those following administration of an equal dosage of DAmB. The levels of ABLC in whole blood were approximately 40 times greater than those in plasma. The ABLC model differed from the DAmB model by (i) a dose- and time-dependent uptake and return between the plasma compartment and apparent cellular components of the blood-sediment compartment and (ii) time-dependent tissue uptake and return to plasma from serially connected compartments. Following infusion of ABLC, there was a nonlinear uptake into the apparent cellular components of the blood-sediment compartment. This uptake was related to the reciprocal of the integral of the total amount of drug infused (i.e., the more drug infused the greater the fractional uptake between 0.5 and 5 mg/kg of body weight for ABLC). The transfer of drug from plasma to the cellular components of the blood-sediment compartment resulted in initial uptake followed by rapid redistribution back to the plasma. The study describes a detailed model of the pharmacokinetics of ABLC and characterizes a potential role of the cellular components of the blood-sediment compartment in the distribution of this new antifungal compound in tissue.


The Melbourne Diabetes Prevention Study (MDPS): study protocol for a randomized controlled trial.
Worldwide, type 2 diabetes (T2DM) prevalence has more than doubled over two decades. In Australia, diabetes is the second highest contributor to the burden of disease. Lifestyle modification programs comprising diet changes, weight loss and moderate physical activity, have been proven to reduce the incidence of T2DM in high risk individuals.As part of the Council of Australia Governments, the State of Victoria committed to develop and support the diabetes prevention program 'Life! Taking action on diabetes' (Life!) which has direct lineage from effective clinical and implementation trials from Finland and Australia. The Melbourne Diabetes Prevention Study (MDPS) has been set up to evaluate the effectiveness and cost-effectiveness of a specific version of the Life! program. We intend to recruit 796 participants for this open randomized clinical trial; 398 will be allocated to the intervention arm and 398 to the usual care arm. Several methods of recruitment will be used in order to maximize the number of participants. Individuals aged 50 to 75 years will be screened with a risk tool (AUSDRISK) to detect those at high risk of developing T2DM. Those with existing diabetes will be excluded. Intervention participants will undergo anthropometric and laboratory tests, and comprehensive surveys at baseline, following the fourth group session (approximately three months after the commencement of the intervention) and 12 months after commencement of the intervention, while control participants will undergo testing at baseline and 12 months only.The intervention consists of an initial individual session followed by a series of five structured-group sessions. The first four group sessions will be carried out at two week intervals and the fifth session will occur eight months after the first group session. The intervention is based on the Health Action Process Approach (HAPA) model and sessions will empower and enable the participants to follow the five goals of the Life! program. This study will determine whether the effect of this intervention is larger than the effect of usual care in reducing central obesity and cardiovascular risk factors and thus the risk of developing diabetes and cardiovascular disease. Also it will evaluate how these two options compare economically. Australian New Zealand Clinical Trials Registry ACTRN12609000507280.


Initiating and Implementing Social Determinants of Health Data Collection in Community Health Centers.
Successfully incorporating social determinants of health (SDH) screening into clinic workflows can help care teams provide targeted care, appropriate referrals, and other interventions to address patients' social risk factors. However, integrating SDH screening into clinical routines is known to be challenging. To achieve widespread adoption of SDH screening, we need to better understand the factors that can facilitate or hinder implementation of effective, sustainable SDH processes. The authors interviewed 43 health care staff and professionals at 8 safety net community health center (CHC) organizations in 5 states across the United States; these CHCs had adopted electronic health record (EHR)-based SDH screening without any external implementation support. Interviewees included staff in administrative, quality improvement, informatics, front desk, and clinical roles (providers, nurses, behavioral health staff), and community health workers. Interviews focused on how each organization integrated EHR-based SDH screening into clinic workflows, and factors that affected adoption of this practice change. Factors that facilitated effective integration of EHR-based SDH screening were: (1) external incentives and motivators that prompted introduction of this screening (eg, grant requirements, encouragement from professional associations); (2) presence of an SDH screening advocate; and (3) maintaining flexibility with regard to workflow approaches to optimally align them with clinic needs, interests, and resources. Results suggest that it is possible to purposefully create an environment conducive to successfully implementing EHR-based SDH screening. Approaching the task of implementing SDH screening into clinic workflows as understanding the interplay of context-dependent factors, rather than following a step-by-step process, may be critical to success in primary care settings.


[Adverse outcome screening in hospitalizations of the Brazilian Unified Health System].
To assess the frequency of screening for potential adverse outcomes in hospitalizations of the Brazilian Unified Health System. A retrospective study, including all hospital admissions of adults in medical clinics (n = 3,565,811) and surgical clinics (n = 2,614,048) in Brazil in 2007. The Hospital Information System was used as a source of information. The measurement of adverse events was based on screening for eleven clinical conditions, as defined by previous international studies, recorded in the secondary diagnosis field. We performed bivariate and multivariate analysis to investigate associations between adverse events, death (dependent variable) and other variables such as age, use of the intensive care unit and performance of surgery. The frequency obtained for both clinic types was 3.6 potential adverse events per 1,000 admissions, with a greater frequency in medical clinics (5.3 per 1,000) than in surgery clinics (1.3 per 1,000). There were differences in the profile of hospital admissions between the two clinics: medical clinics were characterized by a predominance of older adults, longer average length of stay, higher mortality rate and lower total cost of hospitalization. The most common potential adverse outcome was hospital-acquired pneumonia. Cardiac arrest had a higher risk of death (OR= 5.76) compared to other potential adverse outcomes. Increased cost for hospitalizations was associated with sepsis. The conditions used as the screening criteria were associated with greater odds of death even after the introduction of variables such as use of intensive care and surgery. The high frequency of adverse outcomes in hospital admissions indicates a need to develop monitoring strategies and to improve quality of care for improved patient safety.


Superinduction of c-fos by nerve growth factor in the presence of peripherally active benzodiazepines.
Alterations in proto-oncogene expression after stimulation of rat pheochromocytoma (PC12) cells by nerve growth factor (NGF) have been investigated. A specific stimulation of c-fos messenger RNA and protein was detected 30 minutes after treatment. This induction was enhanced more than 100-fold in the presence of peripherally active benzodiazepines. The effect was specific as very little change was observed in the levels of c-rasHa, c-rasKi, c-myc, and N-myc messenger RNA's. Under the conditions used here, NGF treatment ultimately results in neurite outgrowth, with a reduction or cessation of cell division. Thus, stimulation of the c-fos gene in this system appeared to be associated with differentiation and not with cellular proliferation. The effect of benzodiazepines was stereospecific and represents a novel action of these compounds at the level of gene expression.


Predicting Factors of Depression, Antidepressant Use and Positive Response to Antidepressants in Perinatal and Postpartum Women.
In the United States, there is a disparity in knowledge of nationwide depression prevalence, the antidepressant use and the antidepressant responses during perinatal/postpartum periods. This study investigated the predicting factors of depression, antidepressant use and positive antidepressant response during the perinatal/postpartum periods. The 2007-2012 National Health and Nutrition Examination Surveys (NHANES) were combined to identify adult pregnant women, those within the 18-month postpartum period (n=492) and their depression statuses via demographics, health care accessibility, antidepressant use and illicit drug use information. The characteristics of different study groups were compared (depression versus no-depression groups, antidepressant users versus non-antidepressant users, and antidepressant responders versus antidepressant non-responders). Multivariable logistic regression analysis was used to predict factors of perinatal depression (PND)/ postpartum depression (PPD), antidepressant use and antidepressant positive response in PND/PPD. PND/PPD individuals had higher rates of mental health visits. No predicting factor for developing PND/PPD was shown. Antidepressant users were significantly older with insurance and recent health checkups/ mental visits. Being below the poverty level and having some health care accessibility are predictors for being on antidepressants. Recent non-illicit drug use is a predictor for PND/PPD symptom improvement while on antidepressants. The group of those with social-economic disadvantages was more likely to be on antidepressants for PND/PPD. Illicit drug users were less likely to show improvement with antidepressants. The safety and efficacy of antidepressant use during this period is controversial. More studies need to focus on the barriers involving antidepressant treatments, the safety and outcomes of antidepressants for PND/PPD management.


Phlegmonous gastritis associated with the acquired immunodeficiency syndrome/pre-acquired immunodeficiency syndrome.
Unusual infections in the immunocompromised host often present a diagnostic dilemma to the physician. We describe a 29-year-old Haitian man with acquired immunodeficiency syndrome/pre-acquired immunodeficiency syndrome who died of phlegmonous gastritis.


Effects of cone opening, initial moisture content and multiple extrusion on the protein quality of extruded soybean using the Brady Crop Cooker.
In the present study, three cone openings (0.133; 0.106, and 0.080 cm) and three initial moisture content values (9%, 15% and 21%) were used as treatments to evaluate their effects on the protein quality of full-fat soybean flour, extruded in the Brady Crop Cooker. The specific volume, protein and oil contents as well as available lysine content characteristic of the final product, were not affected by the treatments used. Processing temperatures, however, decreased when the initial moisture content of the material was increased. The nitrogen solubility index was affected by the cone opening but not by the moisture content of the material. With respect to the trypsin inhibitors content, the increase in the initial moisture content in soybeans gave conflicting results. At the 21% moisture treatment, the amounts of trypsin inhibitors were higher than those present in the raw material; a similar effect was also observed with urease activity. At the other two moisture contents (9 and 15%) the amounts of trypsin inhibitors and urease activity were decreased by heat treatment, mainly at the 9% moisture level, which were related to the cone opening of the extruder. PER values in rats were influenced by the moisture content and were not affected by the cone opening. Results obtained in the biological assays with chicks, both for weight gain and conversion efficiency, were favored by a decrease in cone opening. Nevertheless, the increase in the moisture content induced a decrease in weight gain at the 5- and 8-week periods, without affecting the conversion efficiency. The effect of consecutive passes of the material through the extruder was also studied. The product obtained with two extrusions presented a good biological value, probably as a consequence of the low values in the trypsin inhibitors and urease activities. When the material was extruded three times, results proved to be poor, due to a reduction to significant low levels of available lysine content--which becomes limiting--, and nitrogen solubility index of the full-fat soybean flour.


Perinatal Immune Activation Produces Persistent Sleep Alterations and Epileptiform Activity in Male Mice.
Increasing evidence suggests a role for inflammation in neuropsychiatric conditions, including autism spectrum disorder (ASD). Previous work in rodents has established that immune activation during critical developmental periods can cause phenotypes that reproduce core features of ASD, including decreased social interaction, aberrant communication, and increased repetitive behavior. In humans, ASD is frequently associated with comorbid medical conditions including sleep disorders, motor hyperactivity, and seizures. Here we use a 'two-hit' immune-activation paradigm to determine whether perinatal immune activation can also produce these comorbid features in mice. In this paradigm, we treated timed-pregnant mice with polyinosinic:polycytidylic acid (Poly I:C), which simulates a viral infection, on gestational day 12.5 according to an established maternal immune activation regimen. A subset of the offspring also received a second 'hit' of lipopolysaccharide (LPS), which simulates a bacterial infection, on postnatal day 9. At 6 weeks of age, mice were implanted with wireless telemetry transmitters that enabled continuous measurements of electroencephalography (EEG), electromyography (EMG), locomotor activity, and subcutaneous temperature. Effects at 7 and 12 weeks of age were compared. Both prenatal Poly I:C and postnatal LPS produced changes in locomotor activity and temperature patterns, increases in slow-wave sleep, and shifts in EEG spectral power, several of which persisted at 12 weeks of age. Postnatal LPS also produced persistent increases in spontaneous bursts of epileptiform activity (spike-wave discharges) that occurred predominantly during sleep. Our findings demonstrate that early-life immune activation can lead to long-lasting physiologic perturbations that resemble medical comorbidities often seen in ASD and other neuropsychiatric conditions.


Assessing evidence supporting redistribution of pressure for pressure ulcer prevention: a review.
The formation and underlying causes of pressure ulcers (PUs) are quite complex, with multiple influencing factors. However, by definition pressure ulcers cannot form without loading, or pressure, on tissue. Clinical interventions typically target the magnitude and/or duration of loading. Pressure magnitude is managed by the selection of support surfaces and postural supports as well as body posture on supporting surfaces. Duration is addressed via turning and weight shifting frequency as well as with the use of dynamic surfaces that actively redistribute pressure on the body surfaces. This article shows that preventative interventions must be targeted to both magnitude and duration and addresses the rationale behind several common clinical interventions--some with more scientific evidence than others.


Requirement for the alternative pathway as well as C4 and C2 in complement-dependent hemolysis via the lectin pathway.
Mannan-binding lectin (MBL) is a C1q-like molecule opsonic for several micro-organisms. MBL can activate C4, C2, and later acting complement components in the presence of serine proteases similar to but distinct from C1r and C1s via the lectin pathway of complement activation. We report here that mannan-coated MBL-sensitized erythrocytes are lysed via the lectin pathway in human serum-Mg-EGTA. The surprising occurrence of MBL-initiated lysis in the absence of calcium contrasts with the calcium requirement for C1q-initiated activation of C4 and C2. C2 is required, and lysis is significantly enhanced when indicator cells presensitized with C4 and then coated with mannan (EAC4-M) are used. The alternative pathway also is required, since lysis is lost when either factor D or factor B is removed and is restored upon reconstitution with the purified protein. Even though MBL is a C-type lectin, it is retained on mannan-coated erythrocytes in the absence of calcium. This contrasts with the absence of calcium-independent retention on mannan immobilized on polystyrene plates or beads, and helps explain the MBL-initiated hemolysis in Mg-EGTA. These investigations show that the alternative pathway as well as C4 and C2 of the classical pathway are required for complement-dependent hemolysis via the lectin pathway and provide a method for assay of lectin pathway-mediated complement activity in human serum that should be useful in unraveling the molecular interactions of this pathway.


Bifidobacterium population analysis in the infant gut by direct mapping of genomic hybridization patterns: potential for monitoring temporal development and effects of dietary regimens.
A bifidobacterial mixed-species microarray platform was used in a proof-of-principle study to address the composition and development of bifidobacteria in DNA extracted from faecal samples. These were collected in a time-course of 2 years since birth and derived from human infants that were breastfed, standard formula-fed or received a prebiotic formula during their weaning period. A set of over 50 samples was analysed, testifying for the throughput of the designed platform for multiple genome hybridizations. The generated data revealed that faecal samples of breastfed infants contained a high abundance of genomic DNA homologous to Bifidobacterium breve. In contrast, faecal samples from standard formula-fed infants lacked detectable amounts of this B. breve DNA but contained genes with high similarity to B. longum. Remarkably, infants that received breastmilk and later a prebiotic formula consisting of a standard formula milk containing a mixture of specific galacto- and fructo-oligosaccharides, continued to harbour a B. breve-dominant faecal population. One infant that received standard formula in combination with the additional B. lactis Bb12 culture, contained significant amounts of faecal DNA belonging to Bb12 but only during the period of ingestion. The microarray platform showed sufficient sensitivity to analyse the B. breve group at the strain level. Overall, the B. breve populations observed in the faecal samples of the studied infants showed a stable composition over time and were unique per infant. In conclusion, our results show the applicability of comparative genome hybridization to study bifidobacterial populations in infant faecal samples without the use of any amplification step.


[Kinetics of sodium current decay during normal axon membrane repolarization and in the presence of scorpion toxin].
Decay of sodium currents in repolarization ("tail current") was studied in from axonal membrane. The decay in the membrane repolarization to -40 divided by -60 mV has two exponential components: fast and slow. The fraction of the slow component in the total "tail current" (theta M) decreases as the repolarization potential (Vp) becomes more negative; at Vp more negative than -80 mV "tail" follows practically one-exponential time course. When lengthening the test pulse (at the given Vp) the fraction of the fast component in the "tail" decreases quicker than that of the slow component, following approximately the kinetics of inactivation during the tests pulse. Scorpion toxin treatment results in slowing down "tail" kinetics mainly at the expense of increasing the fraction of the slow component. A kinetic diagram assuming two open state for the channel is suggested. A hypothesis is advanced that scorpion toxin, DDT and trinitrophenol have a common "site" to interact with the gating mechanism of the sodium channel.


Regulation of brown adipose tissue lipogenesis by thyroid hormone and the sympathetic nervous system.
Thyroid hormone regulates lipogenesis differently in rat liver and brown adipose tissue (BAT). In the hypothyroid state, lipogenesis is suppressed in liver but enhanced in BAT. Here we investigated the mechanisms underlying increased lipogenesis in hypothyroid BAT. Housing the animals at 28 degrees C decreased lipogenesis in hypothyroid BAT to euthyroid levels. Denervation resulted in a 90% reduction in lipogenesis in hypothyroid BAT such that levels were lower than in euthyroid tissue. Thyroid hormone treatment of hypothyroid rats stimulated fatty acid synthesis in denervated BAT, as in liver, but decreased it in intact BAT. Steady-state levels of mRNA encoding acetyl-CoA carboxylase, fatty-acid synthase, and spor 14 were measured in similar animals by Northern analysis. The expression of these mRNAs mirrored the lipogenic data, showing that both thyroid hormone and the sympathetic nervous system work at a pretranslational level in this tissue. These data suggest that the increased BAT lipogenesis found with hypothyroidism is mediated by the sympathetic nervous system to counter the reduction in metabolic rate in these animals.


Effects of captopril on the development of tolerance to guanethidine.
Guanethidine is a sympatholytic antihypertensive drug used for the treatment of severe or resistant hypertension. Previous clinical studies have documented the development of tolerance to guanethidine in humans and suggested that fluid retention or increased sensitivity of arterioles to endogenous catecholamines may be responsible for the tolerance development. In this study we investigated the role of the renin-angiotensin system in the development of tolerance to guanethidine in one-kidney, one-clip renovascular and spontaneously hypertensive rats (SHR) using the angiotensin-converting enzyme inhibitor captopril. In the one-kidney model, captopril (30 mg/(kg X day), orally) had no effect on the development of tolerance to guanethidine (50 mg/(kg X day), i.p.) when the drugs were coadministered for 2 wk. However, captopril enhanced the day 1 blood pressure-lowering effect and prevented the development of tolerance to guanethidine in male SHR. These results suggest that the renin-angiotensin system plays a major role in the development of tolerance to guanethidine in SHR but not in one-kidney, one-clip renal hypertensive rats. Thus, the etiology of the hypertensive state may dictate which physiological compensatory mechanisms are activated after antihypertensive drug administration.


Restoration of competency to stand trial: a training program for persons with mental retardation.
This article describes the development and use of a formal training tool for restoration of competency in clients with mental retardation who are incompetent to stand trial. The program was developed at Eleanor Slater Hospital within the Rhode Island Department of Mental Health, Retardation and Hospitals. This article describes the development of The Slater Method, the training tool format, the procedure for use of the Slater Method, and the duration of treatment to restore competency to stand trial in clients with mental retardation. Although the developmentally disabled population is not limited to persons with mental retardation, we have used the phrase mental retardation instead of developmentally disabled because the judicial system more commonly uses mental retardation.


Identifying teaching in wild animals.
After a long period of neglect, the study of teaching in nonhuman animals is beginning to take a more prominent role in research on social learning. Unlike other forms of social learning, teaching requires knowledgeable individuals to play an active role in facilitating learning by the naive. Casting aside anthropocentric requirements for cognitive mechanisms assumed to underpin teaching in our own species, researchers are now beginning to discover evidence for teaching across a wide range of taxa. Nevertheless, unequivocal evidence for teaching remains scarce, with convincing experimental data limited to meerkats, pied babblers, and tandem-running ants. In this review, our aim is to stimulate further research in different species and contexts by providing conceptual and methodological guidelines for identifying teaching, with a focus on natural populations. We begin by highlighting the fact that teaching is a form of cooperative behavior that functions to promote learning in others and show that consideration of these key characteristics is critical in helping to identify suitable targets for future research. We then go on to discuss potential observational, experimental, and statistical techniques that may assist researchers in providing evidence that the criteria that make up the accepted operational definition of teaching have been met. Supplemental materials for this article may be downloaded from http://lb.psychonomic-journals.org/content/supplemental.


Vectorial effect on the evolution of fractional-order vector vortex beams in a strongly nonlocal nonlinear medium.
Propagation of a vector vortex optical field (VVOF) with both fractional order of polarization topological charge $m$m and fractional order of vortex topological charge $n$n with spatially variant states of polarization (SoP) in a strongly nonlocal nonlinear medium (SNNM) is studied. The optical field always evolves reciprocally with a cycle of stretch and shrink in a SNNM with dark stripes forming at $z=t\pi {z_p}$z=tπzp ($t$t denotes an integer number, and ${{z}_p}$zp is a parameter that depends on the initial power of the VVOF and the material constant associated with the response function), as a result from the coherent superposition of the vortices with different order of topological charges and weighting coefficients. In particular, the conversions between linear and circular polarization components occur during propagation, and the converted SoP distributions in different propagation distances depend closely on the topological charges and the initial powers. The evolutions of the Stokes parameters of the fractional-order VVOF (FO-VVOF) during propagation in a SNNM show that the spatial distributions of different polarization components are closely related to the topological charges, the initial powers and the propagation distances, implying that the FO-VVOF can be regarded as a superposition of two different fractional-order vortices with orthogonal circular polarization components. These results provide new strategies on tailoring polarization states in a structured optical field with fractional topological charges.


Management of etonogestrel subdermal implant-related bleeding.
To determine removal rates for unacceptable bleeding in etonogestrel subdermal implant (ESI) users treated with reassurance alone as well as in those receiving reassurance plus medical interventions including doxycycline and ibuprofen, and to compare removal rates between ESI users treated with reassurance alone with those receiving the medical interventions listed above in addition to reassurance. Medical records of 391 women receiving ESI for contraception within the period June 2007-April 2011 were abstracted for patient characteristics and interventions including reassurance, doxycycline, and ibuprofen. Removal rates were compared between intervention groups using the chi2 test and multivariable logistic regression. Contact for bleeding occurred in 128 women (32.7%). Of those, 83 women (21.2% of entire cohort) requested removalfor bleeding (median, 381 days; range, 29-1,078 days). Of 53 women receiving reassurance alone, 40 (75.5%) requested removal, whereas fewer (15 of 33 [45.5%]) women receiving reassurance plus doxycycline requested removal (p = 0.005), even after adjusting for prior contraception use and body mass index at ESI insertion (p = 0.006). Of 10 women receiving ibuprofen and 19 women receiving doxycycline plus ibuprofen, 7 (70.0%) and 11(57.9%), respectively, requested removal for bleeding. Reassurance plus doxycycline is associated with lower removal rates for ESI-related bleeding than is reassurance alone.


T Cell Responses Induced by Attenuated Flavivirus Vaccination Are Specific and Show Limited Cross-Reactivity with Other Flavivirus Species.
Members of the flavivirus genus share a high level of sequence similarity and often circulate in the same geographical regions. However, whether T cells induced by one viral species cross-react with other related flaviviruses has not been globally addressed. In this study, we tested pools of epitopes derived from dengue (DENV), Zika (ZIKV), Japanese encephalitis (JEV), West Nile (WNV), and yellow fever (YFV) viruses by intracellular cytokine staining (ICS) using peripheral blood mononuclear cells (PBMCs) of individuals naturally exposed to DENV or immunized with DENV (TV005) or YF17D vaccine. CD8 T cell responses recognized epitopes from multiple flaviviruses; however, the magnitude of cross-reactive responses was consistently severalfold lower than those to the autologous epitope pools and was associated with lower expression of activation markers such as CD40L, CD69, and CD137. Next, we characterized the antigen sensitivity of short-term T cell lines (TCL) representing 29 different individual epitope/donor combinations. TCL derived from DENV monovalent vaccinees induced CD8 and CD4 T cells that cross-reacted within the DENV serocomplex but were consistently associated with >100-fold-lower antigen sensitivity for most other flaviviruses, with no cross-recognition of YFV-derived peptides. CD8 and CD4 TCL from YF17D vaccinees were associated with very limited cross-reactivity with any other flaviviruses and in five out of eight cases >1,000-fold-lower antigen sensitivity. Overall, our data suggest limited cross-reactivity for both CD4 and CD8 T cell responses between flaviviruses and have implications for understanding immunity elicited by natural infection and strategies to develop live attenuated vaccines against flaviviral species.IMPORTANCE The envelope (E) protein is the dominant target of neutralizing antibodies for dengue virus (DENV) and yellow fever virus (YFV). Accordingly, several DENV vaccine constructs use the E protein in a live attenuated vaccine format, utilizing a backbone derived from a heterologous flavivirus (such as YF) as a delivery vector. This backbone comprises the nonstructural (NS) and capsid (C) antigens, which are dominant targets of T cell responses. Here, we demonstrate that cross-reactivity at the level of T cell responses among different flaviviruses is very limited, despite high levels of sequence homology. Thus, the use of heterologous flavivirus species as a live attenuated vaccine vector is not likely to generate optimal T cell responses and might thus impair vaccine performance.


[Diagnosis of malaria in non-endemic countries : value, limitations and complementarity of existing methods].
The biological diagnosis of malaria is urgent to avoid rapid and fatal outcome. Every year in France, 5,000 imported malaria cases are observed. Thin stained blood smear microscopical examination remains the reference method of diagnosis; however its performance is linked to the professional competence of the biologists. Thus easier methods have been developed (QBC test). Some of them, limited to the diagnosis of malaria due to Plasmodium falciparum do not require highly skilled personal to perform or interpret (antigen detection on dipsticks, tests Parasight or cardboard, ICT Malaria Pf), but limitations and errors occurred. These different tests must be complementary methods of traditional diagnosis. In association with microscopical examinations, they provide rapid and efficient diagnosis of malaria in non-endemic areas. Relying on our experience, the best association is: QBC + thin blood smear and depending of results antigen detection (ParaSight F, ICT Malaria Pf).


CD98 modulates integrin beta1 function in polarized epithelial cells.
The type II transmembrane protein CD98, best known as the heavy chain of the heterodimeric amino acid transporters (HAT), is required for the surface expression and basolateral localization of this transporter complex in polarized epithelial cells. CD98 also interacts with beta1 integrins resulting in an increase in their affinity for ligand. In this study we explored the role of the transmembrane and cytoplasmic domains of CD98 on integrin-dependent cell adhesion and migration in polarized renal epithelial cells. We demonstrate that the transmembrane domain of CD98 was sufficient, whereas the five N-terminal amino acids of this domain were required for CD98 interactions with beta1 integrins. Overexpression of either full-length CD98 or CD98 lacking its cytoplasmic tail increased cell adhesion and migration, whereas deletion of the five N-terminal amino acids of the transmembrane domain of CD98 abrogated this effect. CD98 and mutants that interacted with beta1 integrins increased both focal adhesion formation and FAK and AKT phosphorylation. CD98-induced cell adhesion and migration was inhibited by addition of phosphoinositol 3-OH kinase (PI3-K) inhibitors suggesting these cell functions are PI3-K-dependent. Finally, CD98 and mutants that interacted with beta1, induced marked changes in polarized renal epithelial cell branching morphogenesis in collagen gels. Thus, in polarized renal epithelial cells, CD98 might be viewed as a scaffolding protein that interacts with basolaterally expressed amino acid transporters and beta1 integrins and can alter diverse cellular functions such as amino acid transport as well as cell adhesion, migration and branching morphogenesis.


Regulatory sequences and virus-cell interactions of JC virus.
A set of recombinant DNAs, representing several isolates of JCV, has been constructed. Some of these cloned DNAs transform nonpermissive primary hamster brain cells. Analysis of the early viral mRNAs and proteins isolated from these transformants revealed similarities with the large and small T messages and proteins of SV40. The region of the JCV genome from 0.63 to 0.73 map units was sequenced by the Maxam-Gilbert technique. This segment of DNA specifies several regulatory elements and the amino-terminal portion of the early viral proteins. Comparisons with the analogous regions in the polyomaviruses SV40 and BKV, confirm the close evolutionary relationship of these three viruses. Similarities include palindromic and symmetrical sequences near the origins of DNA replication, binding sites for the large T proteins, and AT-rich region (the Goldberg-Hogness or TATA box), and a large tanden duplication or triplication to the late side of the replication origin (however, these sequences differ). Homology between the sequences coding for the early proteins is also evident (27 of the first 33 amino acids are shared). Of greater interest are features of the JCV genome which differ from those of other polyomaviruses. Absent in JCV and BKV are sequences which resemble the third T antigen-binding site of SV40. In addition, a set of sequences present in JCV and BKV DNA (33 nucleotides in JCV, 22 nucleotides in BKV), and located near a 17 base pair palindrome shared by all three viruses, is missing in SV40 DNA. Another sequence, GGGXGGAG, which is repeated several times in many polyomaviruses and adenoviruses, and which is thought to play a role in DNA replication and/or transcription, is not found in the JCV sequence presented. Finally, the tandem repeat of JCV, unlike those of BKV and SV40, includes the Goldberg-Hogness sequence.


[Chemotherapy for urological cancer : usefulness and feasibility of current protocols].
In this symposium, we discussed the usefulness and feasibility of current chemotherapy protocols for urological cancers including renal cell carcinoma, urothelial cancer, prostate cancer and testicular cancer. (1) Renal cell carcinoma : Although molecular targeted therapy is now becoming a standard therapy for metastatic renal cell carcinoma, the current situation is somehow confusing due to unknown adverse effects and lack of guidelines for indication of each compound. (2) Urothelial cancer : gemcitabine and cisplatin (GC) therapy is taking over methotrexate, vinblastine, doxorubicin and cisplatin (MVAC) therapy because of less toxicity. We focused on a novel biomarker, hENT1, to predict the anti tumor effect of GC therapy. (3)Prostate cancer : The survival benefit of docetaxel for castration resistant prostate cancer was proven by large clinical trials. We dealt with uracil and tegafur (UFT) therapy which has been widely used in Japan. (4)Testicular cancer : It is needles to say that BEP therapy was established as a standard induction therapy for metastatic testicular cancer. However, regarding salvage chemotherapy, it is still controversial whether vinblastine, ifosphamide and platinum (VIP), paclitaxel, ifosphamide and platinum (TIP) and high dose chemotherapy should be selected.


Effects of tobacco and distraction on sensory and slow cortical evoked potentials during task performance.
Sensory and slow cortical evoked potentials (EP) of female smokers were investigated under two conditions in separate sham and real smoking sessions: a non-distraction (ND) condition consisting of a constant fore-period S1-S2, i.e., a single-choice reaction time situation with a tone (S1)/light (S2) key press sequence, and a task distraction (TD) condition identical to ND with the addition of a short-term memory task consisting of four letters presented within the S1-S2 interval with the requirement that subjects repeat after key press to the light. Two individual EP peaks, N1 and P2, were involved in the amplitude analysis of S1 and S2 stimuli; in addition, the amplitude of the slow contingent negative variation (CNV) developing between S1 and S2 was analyzed. Tobacco significantly altered the amplitude of P2 in response to S1 and CNV amplitudes depending on the pre-experimental expired air carbon monoxide (CO) levels of the smokers. Smokers exhibiting low pre-experimental CO levels evidenced tobacco-induced P2 reductions and CNV increments during the ND condition while subjects exhibiting high pre-experimental CO levels displayed similar cortical changes but in the TD conditions. The results are discussed in relation to the role of nicotine and to improved attention and information processing.


Loss of epidermal hypoxia-inducible factor-1α accelerates epidermal aging and affects re-epithelialization in human and mouse.
In mouse and human skin, HIF-1α is constitutively expressed in the epidermis, mainly in the basal layer. HIF-1α has been shown to have crucial systemic functions: regulation of kidney erythropoietin production in mice with constitutive HIF-1α epidermal deletion, and hypervascularity following epidermal HIF-1α overexpression. However, its local role in keratinocyte physiology has not been clearly defined. To address the function of HIF-1α in the epidermis, we used the mouse model of HIF-1α knockout targeted to keratinocytes (K14-Cre/Hif1a(flox/flox)). These mice had a delayed skin phenotype characterized by skin atrophy and pruritic inflammation, partly mediated by basement membrane disturbances involving laminin-332 (Ln-332) and integrins. We also investigated the relevance of results of studies in mice to human skin using reconstructed epidermis and showed that HIF-1α knockdown in human keratinocytes impairs the formation of a viable reconstructed epidermis. A diminution of keratinocyte growth potential, following HIF-1α silencing, was associated with a decreased expression of Ln-322 and α6 integrin and β1 integrin. Overall, these results indicate a role of HIF-1α in skin homeostasis especially during epidermal aging.


Phosphorylation of CPAP by Aurora-A Maintains Spindle Pole Integrity during Mitosis.
CPAP is required for centriole elongation during S/G2 phase, but the role of CPAP in mitosis is incompletely understood. Here, we show that CPAP maintains spindle pole integrity through its phosphorylation by Aurora-A during mitosis. Depletion of CPAP induced a prolonged delay in mitosis, pericentriolar material (PCM) dispersion, and multiple mitotic abnormalities. Further studies demonstrated that CPAP directly interacts with and is phosphorylated by Aurora-A at serine 467 during mitosis. Interestingly, the dispersal of the PCM was effectively rescued by ectopic expression of wild-type CPAP or a phospho-mimic CPAP-S467D mutant, but not a non-phosphorylated CPAP-S467A mutant. Finally, we found that CPAP-S467D has a low affinity for microtubule binding but a high affinity for PCM proteins. Together, our results support a model wherein CPAP is required for proper mitotic progression, and phosphorylation of CPAP by Aurora-A is essential for maintaining spindle pole integrity.


[Tobacco and morphology: pulmonary diseases].
Tobacco is implicated in multisystemic carcinogenesis through more than fifty identified carcinogenic metabolites that produce mutations responsible for alterations in cell cycle, immune response and endocrine regulation. Is one of nine risk factors identified in one third of cancer deaths together with obesity, sedentary, alcohol consumption, sexual promiscuity, drug addiction, and open and closed air contamination. Answering for cardiovascular diseases as the first cause of death in civilized world, tobacco is also pointed as the major factor implicated in the development of COPD (chronic obstructive pulmonary disease), RB-ILD (respiratory bronchiolitis and interstitial lung disease), DIP (desquamative interstitial pneumonia), bronchiolitis and bronchiolocentric interstitial fibrosis, Langerhans cells histiocytosis, eosinophilic pneumonia, sarcoidosis, epidermoid metaplasia in respiratory epithelium and lung cancer. The chronic tobacco induced inflammatory state is the basis for the acquisition of genetic alterations dependent on the tobacco contaminants.


Recent insights into the pathogenesis and therapeutic targets of chronic liver diseases.
Viral hepatitis, alcohol-associated liver disease (ALD) and non-alcoholic fatty liver disease (NAFLD) are the three major causes of chronic liver diseases, which account for approximately 2 million deaths per year worldwide. The current direct-acting antiviral drugs and vaccinations have effectively reduced and ameliorated viral hepatitis infection, but there are still no effective drug treatments for ALD, NAFLD and liver cancer due to the poor understanding of their pathogenesis. To better understand the pathogenesis, the fifth Chinese American Liver Society/Society of Chinese Bioscientists in America Hepatology Division Annual Symposium, which was held virtually on 21-22 October 2022, focused on the topics related to ALD, NAFLD and liver cancer. Here, we briefly highlight the presentations that focus on the current progress in basic and translational research in ALD, NAFLD and liver cancer. The roles of non-coding RNA, autophagy, extrahepatic signalling, macrophages, etc in liver diseases are deliberated, and the application of single-cell RNA sequencing in the study of liver disease is also discussed.


[DNA compact form. 8. X-ray diffraction study of DNA compact particles, formed in solutions containing polyethylene glycol].
A comparative X-ray study of DNA compact particles, formed in PEG-containing solution from native DNA and from DNA molecules with altered secondary structure was carried out. Low-angle reflections, present in X-ray patterns of compact particles (in powder form) from native DNA, correspond to spacings of 84, 42 and 35 A, while wide-angle reflections correspond to spacings of 12.8; 8.4, 6.0, 4.5, 3.4 A. Low-angle reflections at 84 and 42 A are present also in X-ray patterns of compact particles, formed from DNA molecules with altered secondary structures. These two reflections are believed to be the results of an ordering of DNA molecules within the compact particles. The main features of this ordering appear in the first approximation to be independent on DNA secondary structure. The CD spectra of all types of compact particles, mentioned above, were also studied. It has been shown that the intense negative band (lambda approximately 270 nm) in a CD spectrum appeared only in the case of compact particles, formed from native DNA molecules. The nature of the revealed correlation between the 35 A reflection and the CD negative band is discussed. Data presented in the paper allow one to suppose that the 35 A reflection in X-ray patterns and the CD negative band result from specific interactions between double-stranded DNA fragments spatially brought together in compact particles. Such type of interaction is believed to be characteristic only of native DNA molecules.


Replaced right hepatic artery arising from the gastroduodenal artery: a rare and challenging anatomical variant of the Whipple procedure.
Accurate assessment of the vascular anatomy is a prerequisite of any pancreatic resection, since an unnoticed arterial injury in the context of a complex resection such as Whipple procedure, can seriously jeopardize patient's safety. This article aims to describe an infrequent anatomic variant of a replaced right hepatic artery originating directly from the gastroduodenal artery and its potential implications for duodenopancreatectomy, as the gastroduodenal artery is routinely divided. We present here two different cases of this arterial abnormality identified during a Whipple procedure and its implications in each different setting. Preoperative identification of anatomical variations is essential for proficient surgical planning. Nevertheless, when detected during surgery, an meticulous dissection of the hepatoduodenal ligament is required to identify all the vascular relations in order to avoid irreversible damage.


Ultra-rapid preparation of total genomic DNA from isolates of yeast and mould using Whatman FTA filter paper technology - a reusable DNA archiving system.
Conventional methods for purifying PCR-grade fungal genomic DNA typically require cell disruption (either physical or enzymatic) coupled with laborious organic extraction and precipitation stages, or expensive column-based technologies. Here we present an easy and extremely rapid method of preparing yeast and mould genomic DNAs from living cultures using Whatman FTA filter matrix technology. Aqueous suspensions of yeast cells or hyphal fragments and conidia (in the case of moulds) are applied directly (or after freeze-thawing) to dry FTA filters. Inoculated filters are then subjected to brief microwave treatment, to dry the filters and inactivate the organisms. Filter punches are removed, washed rapidly, dried and placed directly into PCR reactions. We show that this procedure inactivated all of the 38 yeast and 75 mould species tested, and generated PCR-grade DNA preparations in around 15 minutes. A total of 218 out of 226 fungal isolates tested liberated amplifiable DNA after application to FTA filters. Detection limits with yeast cultures were approximately 10 colony-forming units per punch. Moreover, we demonstrate that filter punches can be recovered after PCR, washed and used in fresh PCR reactions without detectable cross-contamination. Whatman FTA technology thus represents a cheap, ultra-rapid method of fungal genomic DNA preparation, and also potentially represents a powerful fungal DNA archiving and storage system.


Doctor-patient relationship: a covenant of trust.
The goals of medicine as a profession dedicated to healing and caring of the sick in a dignified manner depend very much on a stable and trusting doctor-patient relationship. In the last decade, rapid changes in the healthcare delivery system and socio-political climate have resulted in considerable strain on this relationship. What is needed is a reiteration of the fiduciary obligation of the physician and the central role of the patient in the relationship. Concrete steps and changes at the institutional and individual levels need to be taken to preserve the element of trust in the patient-physician covenant. Only then can the medical profession retain the moral dimension of its role in society.


NMR-based discovery of lead inhibitors that block DNA binding of the human papillomavirus E2 protein.
The E2 protein is required for the replication of human papillomaviruses (HPVs), which are responsible for anogenital warts and cervical carcinomas. Using an NMR-based screen, we tested compounds for binding to the DNA-binding domain of the HPV-E2 protein. Three classes of compounds were identified which bound to two distinct sites on the protein. Biphenyl and biphenyl ether compounds containing a carboxylic acid bind to a site near the DNA recognition helix and inhibit the binding of E2 to DNA. Benzophenone-containing compounds which lack a carboxylic acid group bind to the beta-barrel formed by the dimer interface and exhibit negligible effects on the binding of E2 to DNA. Structure-activity relationships from the biphenyl and biphenyl ether compounds were combined to produce a compound [5-(3'-(3",5"-dichlorophenoxy)-phenyl)-2,4-pentadienoic acid] with an IC50 value of approximately 10 microM. This compound represents a useful lead for the development of antiviral agents that interfere with HPV replication and further illustrates the usefulness of the SAR by NMR method in the drug discovery process.


Isolation of koala retroviruses from koalas in Japan.
Koala retrovirus (KoRV) is considered to be associated with leukemia, lymphoma and immunodeficiency-like diseases in koalas. We therefore conducted a pilot study of KoRV infection in five Queensland koalas in Kobe Municipal Oji Zoo. By polymerase chain reaction to detect partial env and pol genes of KoRV in genomic DNA isolated from whole blood and feces, all five koalas were found to be positive for KoRV proviruses. We succeeded in culturing koala lymphocytes from less than 1 ml blood for over 14 days in the presence of recombinant human interleukin-2. By coculturing the lymphocytes with human embryonic kidney (HEK) 293T cells, we isolated KoRVs from all five koalas. We designated these isolates as strains OJ-1 to OJ-5. By electron microscopy, we observed C-type retroviral particles in HEK 293T cells chronically infected with KoRV strain OJ-4. This is the first report on the isolation of KoRV from koalas in a Japanese zoo.


Reflective writing as a tool for assessing teamwork in bioscience: insights into student performance and understanding of teamwork.
To ensure a modern bioscience curriculum that responds to the current needs of stakeholders, there is a need to embed a range of generic capabilities that enables graduates to succeed in and contribute to a rapidly changing world, as well as building strong bioscience skills and knowledge. The curriculum must also prepare students for a rapidly evolving competitive work place and align with the needs of industry. This creates a challenge, how do we develop generic capabilities without losing discipline content. This report analyses teamwork projects embedded in an undergraduate Biotechnology degree designed to promote teamwork skills along with a deeper understanding of the underpinning biochemistry. Student reflective writing was used to capture students' understanding and experience of teamwork as well as provide insight into their metacognition. The analysis demonstrates that 73% of Year 3 and 93% of Year 4 students were capable of learning about teamwork through reflective writing. While the importance of frequent high quality communication was a common theme, evidence suggests that many students were unsophisticated in their use of communication software. The analysis also highlighted the depth of metacognition that underpins successful team function and the significant weaknesses in self-insight some students possess. These findings challenge assumptions regarding student capacity for leadership and the ability of some students to contribute to successful team outcomes. It is essential for the design of teamwork experiences to fully understand the competencies that underlie teamwork, the metacognitive processes required, and ensure that assessments are fair and measure individual academic performance.


Zones of approach for craniofacial resection: minimizing facial incisions for resection of anterior cranial base and paranasal sinus tumors.
Anterior cranial base tumors are surgically resected with combined craniofacial approaches that frequently involve disfiguring facial incisions and facial osteotomies. The authors outline three operative zones of the anterior cranial base and paranasal sinuses in which tumors can be resected with three standard surgical approaches that minimize transfacial incisions and extensive facial osteotomies. The zones were defined by performing dissections on 10 cadaveric heads and by evaluating radiographic images of patients with anterior cranial base tumors. The three approaches performed on each cadaver were transbasal, transmaxillary, and extended transsphenoidal. Three zones of approach were defined for accessing tumors of the anterior cranial base, nasal cavity, and paranasal sinuses. Zone 1 is exposed by the transbasal approach, which is limited anteriorly by the supraorbital rim, posteriorly by the optic chiasm and clivus, inferiorly by the palate, and laterally by the medial orbital walls. This approach allows access to the entire anterior cranial base, nasal cavity, and the majority of maxillary sinuses. The limitation imposed by the orbits results in a blind spot in the superolateral extent of the maxillary sinus. Zone 2 is exposed by a sublabial maxillotomy approach and accesses the entire maxillary sinus, including the superolateral blind spot and the ipsilateral anterior cavernous sinus. However, access to the anterior cranial base is limited. Zone 3 is exposed by the transsphenoidal approach. This approach accesses the midline structures but is limited by the lateral nasal walls and intracavernous carotid arteries. An extended transsphenoidal approach allows further exposure to the anterior cranial base, clivus, or cavernous sinuses. The use of the endoscope facilitates tumor resection in the nasal cavity and paranasal sinuses. The operative zones outlined offer minimally invasive craniofacial approaches to accessing lesions of the anterior cranial base and paranasal sinuses, obviating facial incisions and facial osteotomies. Case illustrations demonstrating the approach selection paradigm are presented.


The production and molecular properties of the zinc beta-lactamase of Pseudomonas maltophilia IID 1275.
The production and purification of a tetrameric zinc beta-lactamase from Pseudomonas maltophilia IID 1275 were greatly improved. Three charge variants were isolated by chromatofocusing. The subunits each contain two atomic proportions of zinc and (in two of the variants) one residue of cysteine. The thiol group is not required for activity, nor does it appear to bind to the metal. Replacement of zinc by cobalt, cadmium or nickel takes place at a measurable rate, and gives enzymes that are less active than the zinc enzyme. The properties of this enzyme differ from those of the other known zinc beta-lactamase, beta-lactamase II from Bacillus cereus. The amino acid sequence of the N-terminal 32 residues was determined; there is no similarity to the N-terminal sequences of other beta-lactamases.


Reaction monitoring via benchtop nuclear magnetic resonance spectroscopy: A practical comparison of on-line stopped-flow and continuous-flow sampling methods.
The ability for nuclear magnetic resonance (NMR) spectroscopy to provide quantitative, structurally rich information makes this spectroscopic technique an attractive reaction monitoring tool. The practicality of NMR for this type of analysis has only increased in the recent years with the influx of commercially available benchtop NMR instruments and compatible flow systems. In this study, we aim to compare 19 F NMR reaction profiles acquired under both on-line continuous-flow and stopped-flow sampling methods, with modern benchtop NMR instrumentation, and two reaction systems: a homogeneous imination reaction and a biphasic activation of a carboxylic acid to acyl fluoride. Reaction trends with higher data density can be acquired with on-line continuous-flow analyses, and this work highlights that representative reaction trends can be acquired without any correction when monitoring resonances with a shorter spin-lattice relaxation time (T1 ), and with the used flow conditions. On-line stopped-flow analyses resulted in representative reaction trends in all cases, including the monitoring of resonances with a long T1 , without the need of any correction factors. The benefit of easier data analysis, however, comes with the cost of time, as the fresh reaction solution must be flowed into the NMR system, halted, and time must be provided for spins to become polarized in the instrument's external magnetic field prior to spectral measurement. Results for one of the reactions were additionally compared with the use of a high-field NMR.


Epigenetic modulation of macrophage polarization- perspectives in diabetic wounds.
Diabetes is a chronic metabolic disorder that poses a global burden to healthcare. Increasing incidence of diabetes-related complications in the affected population includes a delay in wound healing that often results in non-traumatic limb amputations. Owing to the intricacies of the healing process and crosstalk between the multitude of participating cells, the identification of hyperglycaemia-induced changes at both cellular and molecular levels poses a challenge. Macrophages are one of the key participants in wound healing and continue to exert functional changes at the wound site since the time of injury. In the present review, we discuss the role of these cells and their aberrant functions in diabetic wounds. We have extensively studied the process of macrophage polarization (MP) and its modulation through epigenetic modifications. Data from both pre-clinical and clinical studies on diabetes have co-related hyperglycaemia induced changes in gene expression to an increased incidence of diabetic complications. Hyperglycaemia and oxidative stress, create an environment prone to changes in the epigenetic code, that is manifested as an altered inflammatory gene expression. Here, we have attempted to understand the different epigenetic modulations that possibly contribute towards dysregulated MP, resulting in delayed wound healing.


Fish immune response to Myxozoan parasites.
Myxozoan parasites are responsible for important economic losses among fisheries and aquaculture industries, and hence the high interest in studying the immune response of fish against them. The most important data available concerning the immune response of fish against myxosporeans are reviewed, with emphasis on the different innate and adaptive immune mechanisms, their relationship with natural and acquired resistance and the strategies to control and prevent myxosporoses. Cellular effectors (lymphocytes, granulocytes, phagocytes, non-specific cytotoxic cells, rodlet cells) and humoral factors (lysozyme, peroxidades, antiproteases, complement, specific antibodies) have been examined for several myxosporoses, and some immune relevant genes have been studied. This information will be crucial for the future development of vaccines and other preventive strategies such as immunomodulation and selection of disease-resistant strains


Effects of tranexamic acid on coagulofibrinolytic markers during the early stage of severe trauma: A propensity score-matched analysis.
Tranexamic acid (TXA) reduces the risk of bleeding trauma death without altering the need for blood transfusion. We examined the effects of TXA on coagulation and fibrinolysis dynamics and the volume of transfusion during the early stage of trauma. This subanalysis of a prospective multicenter study of severe trauma included 276 patients divided into propensity score-matched groups with and without TXA administration. The effects of TXA on coagulation and fibrinolysis markers immediately at (time point 0) and 3 hours after (time point 3) arrival at the emergency department were investigated. The transfusion volume was determined at 24 hours after admission. TXA was administered to the patients within 3 hours (median, 64 minutes) after injury. Significant reductions in fibrin/fibrinogen degradation products and D-dimer levels from time points 0 to 3 in the TXA group compared with the non-TXA group were confirmed, with no marked differences noted in the 24-hour transfusion volumes between the 2 groups. Continuously increased levels of soluble fibrin, a marker of thrombin generation, from time points 0 to 3 and high levels of plasminogen activator inhibitor-1, a marker of inhibition of fibrinolysis, at time point 3 were observed in both groups. TXA inhibited fibrin(ogen)olysis during the early stage of severe trauma, although this was not associated with a reduction in the transfusion volume. Other confounders affecting the dynamics of fibrinolysis and transfusion requirement need to be clarified.


Addiction research centres and the nurturing of creativity; the Research Institute on Addictions, University at Buffalo.
The Research Institute on Addictions (RIA) was established in 1970 as a research component of the New York State Department of Mental Hygiene. After three decades of serving as a research component of New York State agencies concerned with alcohol and substance abuse, RIA was legislatively transferred to the University at Buffalo in 1999. Today, RIA's cadre of senior research scientists are engaged individually and collaboratively on a multitude of addictions-related studies. The majority of the Institute's ongoing research studies relate to one or more of the following seven broad research domains: causes and consequences of alcohol, marijuana and other drug use; biological and neuroscience; gambling behavior; gender-related studies; dissemination and professional training; treatment; and youth, families and relationships. In this paper, an overview of the structure of the Institute is provided, along with a description of the organizational and scientific culture at RIA. Further information about the Institute, its scientists and its activities can be found at http://www.ria.buffalo.edu.