A uniform-shear rate microfluidic bioreactor for real-time study of proplatelet formation and rapidly-released platelets.
Platelet transfusions, with profound clinical importance in blood clotting and wound healing, are entirely derived from human volunteer donors. Hospitals rely on a steady supply of donations, but these methods are limited by a 5-day shelf life, the potential risk of contamination, and differences in donor/recipient histocompatibility. These challenges invite the opportunity to generate platelets ex vivo. Although much progress has been made in generating large numbers of culture-derived megakaryocytes (Mks, the precursor cells to platelets), stimulating a high percentage of Mks to undergo platelet release remains a major challenge. Recent studies have demonstrated the utility of shear forces to enhance platelet release from cultured Mks. In this study, we performed a computational fluid dynamics (CFD) analysis of several published platelet microbioreactor systems, and used the results to develop a new 7-µm slit bioreactor-with well-defined flow patterns and uniform shear profiles. This uniform-shear-rate bioreactor (USRB-7µm) permits real-time visualization of the proplatelet (proPLT) formation process and the rapid-release of individual platelet-like-particles (PLPs), which has been observed in vivo, but not previously reported for platelet bioreactors. We showed that modulating shear forces and flow patterns had an immediate and significant impact on PLP generation. Surprisingly, using a single flow instead of dual flows led to an unexpected six-fold increase in PLP production. By identifying particularly effective operating conditions within a physiologically relevant environment, this USRB-7µm will be a useful tool for the study and analysis of proPLT/PLP formation that will further understanding of how to increase ex vivo platelet release. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1614-1629, 2017.


Oil droplet interaction with suspended sediment in the seawater column: influence of physical parameters and chemical dispersants.
The interaction of dispersed oil droplets with large diameter suspended particulate materials (SPM) has been little studied. In the current study, particle size, oil characteristics and chemical dispersant significantly influence the adsorption of oil droplets to SPM in seawater. Sediments with a smaller particulate size (clay) approaching that of the oil droplets (2-20 μm) adsorbed more oil per gram than sediments with large particle size (sand). Heavier, more polar oils with a high asphaltene content adsorbed more efficiently to SPM than lighter, less polar oils. A decrease in the smaller, more water soluble oil components in the sediment adsorbed oil was observed for all oil types. Addition of chemical dispersant decreased the adsorption of oil droplets to suspended carbonate sand in an exponential-like manner. No change in the relative distribution of compounds adsorbed to the sediment was observed, indicating dispersants do not alter the dissolution of compounds from oil droplets.


Fatty Acids and Effects on In Vitro and In Vivo Models of Liver Steatosis.
Fatty liver, or steatosis, is a condition of excess accumulation of lipids, mainly under form of triglycerides (TG), in the liver, and it is the hallmark of non-alcoholic fatty liver disease (NAFLD). NAFLD is the most common liver disorder world-wide and it has frequently been associated with obesity, hyperlipidemia and insulin resistance. Free fatty acids (FA) are the major mediators of hepatic steatosis; patients with NAFLD have elevated levels of circulating FA that correlate with disease severity. Steatosis is a reversible condition that can be resolved with changed behaviors, or that can progress towards more severe liver damages such as steatohepatitis (NASH), fibrosis and cirrhosis. In NAFLD, FA of exogenous or endogenous origin accumulate in the hepatocytes and trigger liver damages. Excess TG are stored in cytosolic lipid droplets (LDs) that are dynamic organelles acting as hubs for lipid metabolism. In the first part of this review, we briefly reassumed the main classes of FA and their chemical classification as a function of the presence and number of double bonds, their metabolic pathways and effects on human health. Then, we summarized the main genetic and diet-induced animal models of NAFLD, as well as the cellular models of NAFLD. In recent years, both the diet-induced animal models of NAFLD as well as the cellular models of NAFLD have found ever more application to investigate the mechanisms involved in NAFLD, and we referred to their advantages and disadvantages.


Evaluation of equine digital Starling forces and hemodynamics during early laminitis.
A carbohydrate overload model was used in 8 horses to evaluate Starling forces and hemodynamics of the digit during the prodromal stage of acute laminitis. A pump-perfused extracorporeal digital preparation was used to evaluate blood flow, arterial pressure, venous pressure, capillary pressure, isogravimetric capillary filtration coefficient, osmotic reflection coefficient, and vascular compliance. From these data, pre- and postcapillary resistances and pre- to postcapillary resistance ratios were determined. Vascular and tissue oncotic pressures were estimated from plasma and lymph protein concentrations, respectively. The osmotic reflection coefficient, an estimation of capillary permeability, was determined by means of the lymph protein wash-down technique. Using the collected data, tissue pressure in the digit was calculated by use of the Starling equation. In the isolated digit, mean isogravimetric capillary pressure was 55.13 mm of Hg, mean plasma and lymph oncotic pressures were 22.29 mm of Hg and 7.2 mm of Hg, respectively, the mean osmotic reflection coefficient was 0.66, the mean capillary filtration coefficient was 0.003 ml/min/mm of Hg/100 g, and mean interstitial fluid pressure was 44.82 mm of Hg. The high capillary pressure appeared to be caused by high vascular resistance from the venous side, predisposing to enhanced capillary filtration and interstitial fluid accumulation.


Medication management of depression in the United States and Ontario.
To compare rates of contact for mental problems and receipt of appropriate antidepressant medication management for persons in the general population with major depression in the United States and Ontario, Canada. Survey using the U.S. National Comorbidity Survey and the Mental Health Supplement of the Ontario Health Survey. All persons with major depression as described in DSM-III-R in the previous 12 months, from a multistage random sample of persons aged 21 to 54 years living in households in the United States (n = 574) and Ontario (n = 250) in 1990. Self-reported contact with general medical or mental health specialty providers for mental problems and appropriate medication management, defined as a combination of antidepressant medication use and four or more visits to any provider within the previous 12 months, were the main outcome measures. The proportion of depressed persons receiving appropriate management was lower in the United States than in Ontario (7.3% vs 14.9% in Ontario, adjusted odds ratio [AOR] 95% CI 0.4; 95% confidence interval [CI] 0.2, 0.8). This difference was largely the result of fewer Americans than Canadians having any mental health care from general medical physicians (9.6% in the United States vs 25.8% in Ontario; AOR 0.3; 95% CI 0.1, 0.5) rather than from specialty providers (20.8% in the United States vs 28.9% in Ontario; AOR 0.7; 95% CI 0.4, 1.1). These between-country differences were much greater for the poor than for those with higher incomes. The Ontario-United States AOR of making contact with either type of clinical provider was 7.5 (95% CI 2.7, 20.7) for lowest-income persons but 2.1 (95% CI 0.3, 5.6) for highest-income persons. The proportions of depressed recipients of any mental health care who received appropriate management were similar between countries (23.9% in the United States vs 27.7% in Ontario; AOR 0.8; 95% CI 0.3, 1.7). Most persons with depression in the United States and Ontario do not receive appropriate medication management. The rate of appropriate medication management in the United States relative to Ontario is lower largely because there is less contact with general medical physicians for mental problems, especially for the poor. Economic barriers, rather than knowledge and attitudinal factors, appear to explain this difference.


Invasion of Salmonella enteritidis in avian intestinal epithelial cells in vitro is influenced by short-chain fatty acids.
Fermentation reactions in the caeca of chickens, the predominant place for Salmonella colonization, result in high concentrations of short-chain fatty acids (SCFA). Thus Salmonella bacteria are in close contact with SCFA during their life cycle. A study was carried out to analyse the effects of SCFA on invasion of Salmonella enteritidis in an avian intestinal epithelial cell line. Preincubation of S. enteritidis for 4 h in growth media supplemented with various concentrations of propionate or butyrate resulted in decreased invasion compared to bacteria, preincubated in nonsupplemented media, and to bacteria, preincubated in media supplemented with formate or acetate. Incubation of the S. enteritidis bacteria in media supplemented with mixtures of SCFA mimicking the in vivo caecal concentrations resulted in increased invasion compared with butyrate-exposed bacteria, but equal invasion compared with nonexposed bacteria. Increasing the butyrate concentration in these mixtures did not modify invasion compared with the original mixtures.


The kinetics of proteinase K digestion of linear prion polymers.
Transmissible spongiform encephalopathies such as scrapie are caused by a protein-only infectious agent, known as a prion. It is not clear how a protein can be capable of replicating itself, and the mechanism remains controversial. One influential model hypothesizes that prions are nucleated, macroscopically linear polymers. We investigated the theoretical kinetics of this model and derived predictions which could be used to test the model. In the model, the polymerization and depolymerization rates are independent polymer size. This leads to an exponential size distribution at equilibrium. In agreement with a prediction stemming from this size distribution, the average size of PrP-res polymers was proportional to the square root of the concentration of PrP-res in a published study of in vitro conversion. Prion digestion by proteinase K (PK) is predicted to be biphasic. The second phase of digestion should be virtually independent of the PK concentration and should depend on the initial size distribution of prion polymers. For initially equilibrated polymers with an exponential size distribution, phase two digestion is exponential at a predicted rate. This rate varies in a defined way with the concentration used for equilibration and with other parameters which affect the average polymer size.


The unrecognized potential of pet cats for studying aging and age-related diseases.
Old cats develop chronic diseases similar to diseases in older people. One-fourth of American households own cats, and almost half are more than 7 years old. Cats share the same environment and are exposed to many of the same chemical stresses. In addition, genomic diversity and population stratification are similar to that occurring in people. With these comparative features, the aging cat represents a geroscience model to investigate the pathogenesis and therapeutic interventions for aging. However, cats are generally not recognized as a translational model for aging research mainly because of the lack of knowledge and appreciation within the scientific community. In addition, cat owners are not aware of any research programs designed to enhance healthy aging in their pets because none exist. Much work is needed to inform and educate the scientific community as well as cat owners about the power of aging cats as a transformative model to investigate aging and age-related diseases that will benefit both human and feline health.


CaO-based CO2 sorbents: from fundamentals to the development of new, highly effective materials.
The enormous anthropogenic emission of the greenhouse gas CO2 is most likely the main reason for climate change. Considering the continuing and indeed growing utilisation of fossil fuels for electricity generation and transportation purposes, development and implementation of processes that avoid the associated emissions of CO2 are urgently needed. CO2 capture and storage, commonly termed CCS, would be a possible mid-term solution to reduce the emissions of CO2 into the atmosphere. However, the costs associated with the currently available CO2 capture technology, that is, amine scrubbing, are prohibitively high, thus making the development of new CO2 sorbents a highly important research challenge. Indeed, CaO, readily obtained through the calcination of naturally occurring limestone, has been proposed as an alternative CO2 sorbent that could substantially reduce the costs of CO2 capture. However, one of the major drawbacks of using CaO derived from natural sources is its rapidly decreasing CO2 uptake capacity with repeated carbonation-calcination reactions. Here, we review the current understanding of fundamental aspects of the cyclic carbonation-calcination reactions of CaO such as its reversibility and kinetics. Subsequently, recent attempts to develop synthetic, CaO-based sorbents that possess high and cyclically stable CO2 uptakes are presented.


Effectiveness of the trivalent MF59 adjuvated influenza vaccine in preventing hospitalization due to influenza B and A(H1N1)pdm09 viruses in the elderly in Italy, 2017 - 2018 season.
Background: Evidence on influenza vaccine effectiveness (VE) in preventing mortality and morbidity in the elderly is weak. Our aim was to measure the VE against severe outcomes in the elderly. Methods: We conducted a multicentre hospital-based test-negative design (TND) case-control study, during the 2017/18 season, in four Italian hospitals. The study population included individuals aged ≥65 years hospitalized with Severe Acute Respiratory Infections (SARI). Patients were classified as cases and controls based on the results of the PCR influenza testing. We estimated VE by virus subtypes and specific VE for the trivalent adjuvanted vaccine (TIVadj). Results: 502 patients with SARI were enrolled: 118 (23.5%) tested positive (cases) and 384 (76.5%) tested negative (controls) for influenza. The adjusted VE of 48.5% for all vaccines was comparable to the adjusted VE for the TIVadj vaccine (48.3%). Adjusted VE for the TIVadj vaccine was 67.5% for A(H1N1)pdm09 and 44.5% for B viruses. Conclusion: We show a moderate adjusted VE of the TIVadj against all viruses, a good adjusted VE against A(H1N1)pdm09 strains and a moderate adjusted VE against B strains, despite a mismatch between the B circulating lineage and the lineage included in the vaccine. This is likely due to the cross-protection among B strains induced by the TIVadj in elderly patients.


Comparison of Propofol, Etomidate, and Thiopental in Anesthesia for Electroconvulsive Therapy: A Randomized, Double-blind Clinical Trial.
This study aimed to compare the effects of propofol, thiopental, and etomidate, which are routinely used in anesthesia for electroconvulsive therapy (ECT), on the cardiovascular system, seizure variables, recovery, cognitive functions, and response to treatment. Male patients hospitalized at the Seventh Psychiatry Clinics of the Bakırköy Teaching Hospital for Psychiatry, Neurology, and Neurosurgery who were treated with ECT were investigated prospectively. The effects on cardiovascular system parameters (heart rate, blood pressure, and blood oxygenation), seizure variables (duration and intensity of seizure), and recovery variables were recorded at every session, on prespecified time points, and the findings of the first session were used in this evaluation. In addition, clinical responses to treatment were evaluated with tests of cognitive functions before and after a course of ECT. Adverse effects were recorded. The sociodemographic characteristics of the 3 treatment groups were similar. There were no significant differences among the groups in terms of effects on cardiovascular system variables, seizure variables, and cognitive functions. The clinical response to ECT was good in all groups, without any significant differences. Propofol, etomidate, and thiopental are associated with similar safety and efficacy profiles.


Green technologies--assumption of economic recovery.
Green technologies include implementation of technological projects in the field of environmental protection through all associated components, such as: waters, soil, air or biodiversity. Hence, such projects potentially become a driving force of new economic momentum in the conditions of post-crisis recovery. In addition, the support of this segment by the institutions of the European Union, either in terms of organization through the establishment of rules and systems for monitoring and control of environmental protection measures, and most importantly in terms of finances, by supporting the development of infrastructure for environmental protection, is today an indisputable category. The aim of the research is to show the potential of green technologies in the initiation of economic activities based on content analysis of the collected literature, as well as to determine the correlation between green technologies and environmental protection and the measures for the reduction of the impact of energy sector on the greenhouse gas emissions.


Medicinal Herbs Used in Traditional Management of Breast Cancer: Mechanisms of Action.
Background: Breast cancer is one of the principal causes of death among women and there is a pressing need to develop novel and effective anti-cancer agents. Natural plant products have shown promising results as anti-cancer agents. Their effectiveness is reported as decreased toxicity in usage, along with safety and less recurrent resistances compared with hormonal targeting anti-cancer agents. Methods: A literature search was conducted for all English-language literature published prior to June 2020. The search was conducted using electronic databases, including PubMed, Embase, Web of Science, and Cochrane Library. The search strategy included keywords such as breast cancer, herbs, anti-cancer biologically active components, clinical research, chemotherapy drugs amongst others. Results: The literature provides documented evidence of the chemo-preventative and chemotherapeutic properties of Ginseng, garlic (Allium sativum), Black cohosh (Actaea racemose), Tumeric (Curcuma longa), Camellia sinenis (green tea), Echinacea, Arctium (burdock), Flaxseed (Linum usitatissimum) and Black Cumin (Nigella sativa). Conclusions: The nine herbs displayed anti-cancer properties and their outcomes and mechanisms of action include inhibition of cell proliferation, angiogenesis and apoptosis as well as modulation of key intracellular pathways. However, more clinical trials and cohort human studies should be conducted to provide key evidence of their medical benefits.


Improving smoking cessation after myocardial infarction by systematically implementing evidence-based treatment methods.
We compared the odds of smoking cessation at 2-months post-myocardial infarction (MI), before and after implementing routines optimizing use of evidence-based smoking cessation methods, with start during admission. The following routines were implemented at six Swedish hospitals: cardiac rehabilitation nurses offering smokers consultation during admission, optimizing nicotine replacement therapy and varenicline prescription, and contacting patients by telephone during the 1st week post-discharge. Using logistic regression, odds for smoking cessation at 2-months before (n smokers/n admitted = 188/601) and after (n = 195/632) routine implementation were compared. Secondary outcomes included adherence to implemented routines and assessing the prognostic value of each routine on smoking cessation. After implementation, a larger proportion of smokers (65% vs. 54%) were abstinent at 2-months (OR 1.60 [1.04-2.48]). Including only those counselled during admission (n = 98), 74% were abstinent (2.50 [1.42-4.41]). After implementation, patients were more often counselled during admission (50% vs. 6%, p < 0.001), prescribed varenicline (23% vs. 7%, p < 0.001), and contacted by telephone post-discharge (18% vs. 2%, p < 0.001). Being contacted by telephone post-discharge (adjusted OR 2.74 [1.02-7.35]) and prescribed varenicline (adjusted OR 0.39 [0.19-0.83]) predicted smoking cessation at 2-months. In conclusion, readily available methods for aiding smoking cessation can be implemented effectively in routine practice, with beneficial effects for post-MI patients.


Magnetic Plasmon Networks Programmed by Molecular Self-Assembly.
Nanoscale manipulation of magnetic fields has been a long-term pursuit in plasmonics and metamaterials, as it can enable a range of appealing optical properties, such as high-sensitivity circular dichroism, directional scattering, and low-refractive-index materials. Inspired by the natural magnetism of aromatic molecules, the cyclic ring cluster of plasmonic nanoparticles (NPs) has been suggested as a promising architecture with induced unnatural magnetism, especially at visible frequencies. However, it remains challenging to assemble plasmonic NPs into complex networks exhibiting strong visible magnetism. Here, a DNA-origami-based strategy is introduced to realize molecular self-assembly of NPs forming complex magnetic architectures, exhibiting emergent properties including anti-ferromagnetism, purely magnetic-based Fano resonances, and magnetic surface plasmon polaritons. The basic building block, a gold NP (AuNP) ring consisting of six AuNP seeds, is arranged on a DNA origami frame with nanometer precision. The subsequent hierarchical assembly of the AuNP rings leads to the formation of higher-order networks of clusters and polymeric chains. Strong emergent plasmonic properties are induced by in situ growth of silver upon the AuNP seeds. This work may facilitate the development of a tunable and scalable DNA-based strategy for the assembly of optical magnetic circuitry, as well as plasmonic metamaterials with high fidelity.


[The criteria of cardiac muscle tissue viability under experimental and clinical conditions].
Comparative analysis of cardiac muscular tissue reactivity in myocardial hypertrophy development in experimental and clinical conditions was performed. Stereotyped reactions to overload were distinguished at intracellular, cellular and tissue levels. Cardiac muscular tissue vital activity was estimated through criteria of "chronos", "morphos", "quantum" and "turnover". It was shown that disruption of genetically determined correlation between processes of proliferation, differentiation and death of cardiomyocytes was a common regularity of cardiomyogenesis in formation of myocardial hypertrophy.


[Evaluation of the Extra Short Musculoskeletal Function Assessment questionnaire XSMFA-D in patients with musculoskeletal disorders and surgical or medical in-patient treatment].
The present study had the objective of evaluating the psychometric characteristics of the shortened 16-item version of the German Short Musculoskeletal Function Assessment questionnaire (XSMFA-D), which was designed for routine assessment of functional capacity in patients with various orthopaedic disorders treated either surgically or medically. A total of 382 patients from seven different samples with either osteoarthritis of the knee, osteoarthritis of the hip, rheumatoid arthritis or rotator cuff tear treated either in surgical hospitals or rehabilitation settings were assessed both before and after treatment. The XSMFA-D was compared with both questionnaires measuring similar constructs and widely accepted indicators of health status in musculoskeletal disorders. Psychometric characteristics were computed. The internal consistency (Cronbach's alpha) exceeded 0.90 in most cases for the function index and was between 0.80 and 0.90 in most cases for the bother index of the XSMFA-D. Retest-reliability was between 0.80 and 0.95 for the function index and between 0.60 and 0.92 for the bother index. Correlations between the XSMFA-D subscales and the other questionnaires were substantial, showing construct validity. Criterion validity was also demonstrated as significant relationships with accepted external parameters such as function tests, judgements by physicians, pain self-ratings by patients and disease severity scores were found. Sensitivity to change was as high as for the other assessment instruments used in this study. The XSMFA-D, a very short assessment instrument, proved to be practical, reliable, valid and sensitive to change among the various patient samples studied. It may be recommended for the assessment of patients' state as well as the evaluation of treatments.


The Use of Macro, Micro, and Trace Elemental Profiles to Differentiate Commercial Single Vineyard Pinot noir Wines at a Sub-Regional Level.
The compositional authentication of wine is of great interest, as the geographic origin of the grapes is often associated with quality, uniqueness, and authenticity. Previous elemental fingerprinting studies mainly discriminated wines from different countries or regions within a country. Here, we report the use of element profiles to distinguish commercial Pinot noir wines from five sub-regional appellations or neighborhoods within one American viticultural area (AVA). Fifty-three single cultivar wines were collected over two harvests and analyzed using microwave plasma-atomic emission spectroscopy (MP-AES) and inductively coupled plasma-mass spectrometry (ICP-MS). Of 62 monitored elements that were quantified with fully validated methods, 24 and 32 elements differed significantly across the neighborhoods and vintages, respectively (p < 0.05). Targeted canonical variate analysis (CVA) explained 85%-90% of the variance ratio across the two vintages, indicating persistent and stable elemental fingerprints of wines at a sub-regional level. A sixth, newly founded neighborhood was correctly grouped separately from the others using a Soft Independent Modeling of Class Analogy (SIMCA), indicating the potential of elemental fingerprints for wine authenticity.


Evaluation and management of joint pain.
The purpose of this article was to provide a review for orthopaedic nurses and nurse practitioners who evaluate, manage, and care for patients with joint pain. Joint pain is a common complaint evaluated by primary care providers. The causation of joint pain is complicated to identify because of an extensive range of differential diagnosis. The history and physical examination are crucial components in evaluating and managing joint pain. The primary care provider uses clinical factors such as patient demographics, presence of inflammation, acute/chronic duration, extra-articular manifestations, pattern of joint involvement, and disease chronology. Many rheumatologic laboratory tests are nonspecific, but aspiration of the joint with synovial fluid analysis may provide diagnostic clues, especially to differentiate infection versus inflammation. Primary care providers utilize both pharmacologic and nonpharmacologic regimens to manage acute and chronic joint pain.


MicroRNA-138 regulates hypoxia-induced endothelial cell dysfunction by targeting S100A1.
The Ca(2+) sensor S100A1 is essential for proper endothelial cell (EC) nitric oxide (NO) synthase (eNOS) activation. S100A1 levels are greatly reduced in primary human microvascular ECs subjected to hypoxia, rendering them dysfunctional. However mechanisms that regulate S100A1 levels in ECs are unknown. Here we show that ECs transfected with a S100A1-3' untranslated region (UTR) luciferase reporter construct display significantly reduced gene expression when subjected to low oxygen levels or chemical hypoxia. Bioinformatic analysis suggested that microRNA -138 (MiR-138) could target the 3'UTR of S100A1. Patients with critical limb ischemia (CLI) or mice subjected to femoral artery resection (FAR) displayed increased MiR-138 levels and decreased S100A1 protein expression. Consistent with this finding, hypoxia greatly increased MiR-138 levels in ECs, but not in skeletal muscle C2C12 myoblasts or differentiated myotubes or primary human vascular smooth muscle cells. Transfection of a MiR-138 mimic into ECs reduced S100A1-3 'UTR reporter gene expression, while transfection of an anti MiR-138 prevented the hypoxia-induced downregulation of the reporter gene. Deletion of the 22 nucleotide putative MiR-138 target site abolished the hypoxia-induced loss of reporter gene expression. Knockdown of Hif1-α mediated by siRNA prevented loss of hypoxia-induced reporter gene expression. Conversely, specific activation of Hif1-α by a selective prolyl-hydroxylase inhibitor (IOX2) reduced reporter gene expression even in the absence of hypoxia. Finally, primary ECs transfected with a MiR-138 mimic displayed reduced tube formation when plated onto Matrigel matrix and expressed less NO when stimulated with VEGF. These effects were reversed by gene transfer of S100A1 using recombinant adenovirus. We conclude that hypoxia-induced MiR-138 is an essential mediator of EC dysfunction via its ability to target the 3'UTR of S100A1.


Cell proliferation and apoptosis in the exocrine pancreas of azaserine-treated rats and N-nitrosobis(2-oxopropyl)amine-treated hamsters.
Altered cell proliferation is an important characteristic of neoplastic development, hence the quantitation of cell growth in normal and (pre)neoplastic pancreatic tissue will provide useful information on the process (and modulation) of pancreatic tumour promotion in experimental animals. Cell proliferation was estimated by means of BrdU-incorporation in normal pancreatic tissue and in putative preneoplastic pancreatic tissue of azaserine-treated rats and of N-nitrosobis(2-oxopropyl)amine (BOP)-treated hamsters at 2, 6, 12, 26 and 52 weeks post-treatment. Furthermore, the effects of diets high in vegetable oil or fish oil on the BrdU labeling index, the Ki-67 labeling index as well as the apoptotic labeling index were determined in azaserine-induced atypical acinar cell foci (AACF) in the pancreas of rats. In saline- and azaserine-treated rats, the BrdU labeling index (LI) in normal pancreatic acinar cells showed a similar time-related decrease. In azaserine-treated rats, the BrdU-LI in AACF was significantly higher than in normal acinar tissue. The BrdU, Ki-67 and apoptotic labeling indices in pancreatic AACF of rats fed diets high in vegetable oil versus fish oil were 24.8+/-1.7 vs 15.5+/-1.2 (BrdU; P<0.05), 9.9+/-0.8 vs 6.5+/-0.5 (Ki-67; P<0.001) and 1.31+/-0.12 vs 0.97+/-0.11 (apoptosis; P<0.05), respectively. In hamsters, up to 26 weeks after BOP-treatment the BrdU-LI in acinar cells was significantly higher than in saline-treated animals. In BOP-treated hamsters, ductal cells showed a significantly increased LI 2 weeks post-treatment, whereas the LI in centroacinar cells had significantly increased 2 and 26 weeks post-treatment. In hamster pancreas the LI was significantly higher in tubular ductal complexes than in cystic ductal complexes and was highest in borderline lesions. It is concluded that (i) determination of BrdU-LI provides a reliable parameter to discriminate between putative preneoplastic lesions with a high or low growth potential, hence with a high or low potential to develop into ultimate carcinomas, (ii) BOP causes increased DNA synthesis in normal acinar cells, leading to increased risk of additional DNA damage, implicating the acinar cell to be putatively involved in the development of ductular adenocarcinomas in the hamster pancreas and (iii) quantitation of cell proliferation and apoptosis may be useful to determine the enhancing or inhibitory effects of life-style factors on pancreatic carcinogenesis.


Breaking bad news issues: a survey among radiation oncologists.
Discussion of bad news and resuscitation in terminal cancer is an important but difficult and often neglected issue in day-to-day oncology practice. We interviewed 35 radiation oncologists using an indigenous 15-item questionnaire on their beliefs about breaking bad news and resuscitation to terminal cancer patients. Most responders had an oncology experience of three to seven years (20/35). Thirty-two were comfortable discussing cancer diagnosis, prognosis and life expectancy-related issues. A similar number believed all cancer-related information should be disclosed, while only four believed in imparting all information in one visit. All agreed that disclosing sensitive information did not affect survival. When requested by relatives to withhold truth from patients, 11 said they would not comply, 22 agreed to tell the truth only if asked and two agreed to avoid difficult questions. Twenty responders denied having been adequately trained in breaking bad news and were keen on dedicated classes or sessions in this area of practice. Most (33/35) believed that Indian patients were keen on knowing their diagnosis and prognosis. Although all agreed to the importance of discussing resuscitation, only 17 believed patients should be involved. Majority (20/35) agreed that the issue needs to be discussed while the patient was conscious. Patients with unsalvageable disease were deemed unsuitable for aggressive resuscitation by 30 responders while the rest believed it should be offered to all. Most (21/35) admitted to feeling depressed after breaking bad news though only seven felt disclosure was more stressful than untruthful statements. Only four knew of a law regarding resuscitation in cancer. Observing the widely varied beliefs and practices for disclosing bad news, it is recommended that such training be a regular part of medicine curriculum, especially in the Oncology setting.


Mass media campaigns designed to support new pictorial health warnings on cigarette packets: evidence of a complementary relationship.
In Australia, introduction of pictorial health warnings on cigarette packets was supported by a televised media campaign highlighting illnesses featured in two of the warning labels--gangrene and mouth cancer. Two studies examined whether the warnings and the television advertisements complemented one another. Population telephone surveys of two cross-sections of adult smokers measured changes in top-of-mind awareness of smoking-related health effects from before (2005; n=587) to after the pack warnings were introduced (2006; n=583). A second study assessed cognitive and emotional responses and intentions to quit after smokers watched one of the campaign advertisements, comparing outcomes of those with and without prior pack warning exposure. Between 2005 and 2006, the proportion of smokers aware that gangrene is caused by smoking increased by 11.2 percentage points (OR=23.47, p=0.000), and awareness of the link between smoking and mouth cancer increased by 6.6 percentage points (OR=2.00, p=0.006). In contrast, awareness of throat cancer decreased by 4.3 percentage points, and this illness was mentioned in the pack warnings but not the advertisements. In multivariate analyses, smokers who had prior exposure to the warnings were significantly more likely to report positive responses to the advertisements and stronger post-exposure quitting intentions. Television advertisements and pictorial health warnings on cigarette packets may operate in a complementary manner to positively influence awareness of the health consequences of smoking and motivation to quit. Jurisdictions implementing pictorial warnings should consider the benefits of supportive mass media campaigns to increase the depth, meaning and personal relevance of the warnings.


Streamlining the Assessment of Patient-Reported Outcomes in Weight Loss and Body Contouring Patients: Applying Computerized Adaptive Testing to the BODY-Q.
The BODY-Q is a widely used patient-reported outcome measure of surgical outcomes in weight loss and body contouring patients. Reducing the length of the BODY-Q assessment could overcome implementation barriers in busy clinics. A shorter BODY-Q could be achieved by using computerized adaptive testing, a method to shorten and tailor assessments while maintaining reliability and accuracy. In this study, the authors apply computerized adaptive testing to the BODY-Q and assess computerized adaptive testing performance in terms of item reduction and accuracy. Parameters describing the psychometric properties of 138 BODY-Q items (i.e., questions) were derived from the original validation sample (n = 734). The 138 items are arranged into 18 scales reflecting Appearance, Quality of Life, and Experience of Care domains. The authors simulated 1000 administrations of the computerized adaptive testing until a stopping rule, reflecting assessment accuracy of standard error less than 0.55, was met. The authors describe the reduction of assessment length in terms of the mean and range of items administered. The authors assessed accuracy by determining correlation between full test and computerized adaptive testing scores. The authors ran 54 simulations. Mean item reduction was 36.9 percent (51 items; range, 48 to 138 items). Highest item reduction was achieved for the Experience of Care domain (56.2 percent, 22.5 items). Correlation between full test scores and the BODY-Q computerized adaptive test scores averaged 0.99. Substantial item reduction is possible by using BODY-Q computerized adaptive testing. Reduced assessment length using BODY-Q computerized adaptive testing could reduce patient burden while preserving the accuracy of clinical patient-reported outcomes for patients undergoing weight loss and body contouring operations.


Z-form DNA specific binding geometry of Zn(II) meso-tetrakis(N-methylpyridinium-4-yl)porphyrin probed by linear dichroism spectroscopy.
Zn(II) meso-tetrakis(N-methylpyridinium-4-yl)porphyrin (ZnTMPyP) produced a unique linear dichroism (LD) spectrum when forming a complex with Z-form poly[d(G-C)(2)]. The spectrum was characterized by a large positive wavelength-dependent LD signal in the Soret absorption region. The magnitudes of LD in both the DNA and Soret band increased as the [porphyrin]/[DNA base] ratio increased and were larger by 20-40 times compared to the negative LD of the ZnTMPyP bound to B-form poly[d(G-C)(2)] and poly[d(A-T)(2)]. The angles calculated from LD were respectively 49° and 42° for B(x) and B(y) transitions of the porphyrin with respect to the local helix axis of Z-form poly[d(G-C)(2)]. The appearance of the unique LD spectrum for the Z-form poly[d(G-C)(2)] complex was accompanied by a bisignate circular dichroism spectrum in the Soret region, whose magnitude was proportional to the square of the porphyrin concentration, suggesting a stacking interaction between Z-form poly[d(G-C)(2)]-bound ZnTMPyP with other bound ZnTMPyP. From these observations, a conceivable binding mode of ZnTMPyP to Z-form poly[d(G-C)(2)] complex was proposed, in which ZnTMPyP binds at the major groove or across the groove. In contrast with Z-form poly[d(G-C)(2)], ZnTMPyP binds to poly[d(A-T)(2)] in a monomeric manner with the angles of 57° and 59° for the two porphyrin's transition moments with respect to the local polynucleotide helix axis. The polarized spectral properties of ZnTMPyP bound to B-form poly[d(G-C)(2)] coincide with the intercalated nonmetallic TMPyP, namely, a negative CD signal in the Soret band and a negative wavelength-dependent reduced LD signal, with a magnitude larger than that in the DNA absorption region in spite of its axial ligands.


Introduction of nursing informatics in the nursing baccalaureate program at the American University of Beirut.
The rapid expansion of computer use for various nursing activities has made computer technology an important part of the curriculum in many schools of nursing across the world. Computer technology has become an alternative tool for teaching practice skills to students in a variety of educational settings and disciplines. Information technology (IT) is not yet well recognized in the Lebanese nursing curricula. This article traces the events of planning, developing, integrating, and evaluating nursing informatics on the Bachelor of Science in Nursing (BSN) Program of the School of Nursing (SON), at The American University of Beirut (AUB), Lebanon.


Transgelin, a p53 and PTEN-Upregulated Gene, Inhibits the Cell Proliferation and Invasion of Human Bladder Carcinoma Cells in Vitro and in Vivo.
Transgelin (TAGLN/SM22-α) is a regulator of the actin cytoskeleton, affecting the survival, migration, and apoptosis of various cancer cells divergently; however, the roles of TAGLN in bladder carcinoma cells remain inconclusive. We compared expressions of TAGLN in human bladder carcinoma cells to the normal human bladder tissues to determine the potential biological functions and regulatory mechanisms of TAGLN in bladder carcinoma cells. Results of RT-qPCR and immunoblot assays indicated that TAGLN expressions were higher in bladder smooth muscle cells, fibroblast cells, and normal epithelial cells than in carcinoma cells (RT-4, HT1376, TSGH-8301, and T24) in vitro. Besides, the results of RT-qPCR revealed that TAGLN expressions were higher in normal tissues than the paired tumor tissues. In vitro, TAGLN knockdown enhanced cell proliferation and invasion, while overexpression of TAGLN had the inverse effects in bladder carcinoma cells. Meanwhile, ectopic overexpression of TAGLN attenuated tumorigenesis in vivo. Immunofluorescence and immunoblot assays showed that TAGLN was predominantly in the cytosol and colocalized with F-actin. Ectopic overexpression of either p53 or PTEN induced TAGLN expression, while p53 knockdown downregulated TAGLN expression in bladder carcinoma cells. Our results indicate that TAGLN is a p53 and PTEN-upregulated gene, expressing higher levels in normal bladder epithelial cells than carcinoma cells. Further, TAGLN inhibited cell proliferation and invasion in vitro and blocked tumorigenesis in vivo. Collectively, it can be concluded that TAGLN is an antitumor gene in the human bladder.


There is more to biological behavior than causation and control.
Biorobots may model the causation of relatively simple behaviors, but many animal behaviorists are concerned with complex cognitive traits and their evolution. Biorobotics seems limited in its ability to model cognition and to provide evolutionary explanations. Also, if robots could model complex traits, such as theory of mind, underdetermination could be problematic. Underdetermination is also a challenge for comparative psychologists.


Co-expression of the hepatitis B surface and core antigens using baculovirus multiple expression vectors.
The hepatitis B (HB) virus DNA sequences coding for the pre-core (preC) or C antigens (HBpcAg, HBcAg) have been inserted into the baculovirus plasmid transfer vector, pAcYM1, such that the HB viral sequences are under the control of the polyhedrin promoter of Autographa californica nuclear polyhedrosis virus (AcNPV). Spodoptera frugiperda cells infected with either of the derived recombinant plasmids in the presence of infectious AcNPV DNA yielded recombinant, polyhedrin-negative viruses that expressed high levels of the respective HBpcAg or HBcAg (representing approx. 5 to 10% and approx. 40% of the stained cellular proteins, respectively). The particulate 27 nm HBcAgs have been purified to homogeneity from infected cell extracts by density gradient centrifugation. Dual expression transfer vectors containing the HBcAg gene sequences and the coding sequences of the HB viral S antigen (HBsAg), each gene under the control of its own copy of the polyhedrin promoter, have also been constructed and used to derive recombinant viruses. The recombinant with the HB C and S genes expressed high levels of the HBcAg (approx. 40% of the cellular proteins) and low levels of the HBsAg (approx. 2% of the stained cellular proteins). Dual expression, occluded, recombinant baculoviruses that make HBsAg, as well as the AcNPV polyhedrin protein, have been prepared that are highly infectious for Trichoplusia ni caterpillars, allowing reproducible preparation of the antigen in larvae. Using radioimmunoassays (RIAs) and ELISAs, the recombinant HBcAg (RIA) and HBsAg (ELISA) have been used to identify human antibodies to HB virus with results that compare favourably with the data obtained with non-recombinant antigens.


Characterizing COVID-19 and Influenza Illnesses in the Real World via Person-Generated Health Data.
The fight against COVID-19 is hindered by similarly presenting viral infections that may confound detection and monitoring. We examined person-generated health data (PGHD), consisting of survey and commercial wearable data from individuals' everyday lives, for 230 people who reported a COVID-19 diagnosis between March 30, 2020, and April 27, 2020 (n = 41 with wearable data). Compared with self-reported diagnosed flu cases from the same time frame (n = 426, 85 with wearable data) or pre-pandemic (n = 6,270, 1,265 with wearable data), COVID-19 patients reported a distinct symptom constellation that lasted longer (median of 12 versus 9 and 7 days, respectively) and peaked later after illness onset. Wearable data showed significant changes in daily steps and prevalence of anomalous resting heart rate measurements, of similar magnitudes for both the flu and COVID-19 cohorts. Our findings highlight the need to include flu comparator arms when evaluating PGHD applications aimed to be highly specific for COVID-19.


Identification, pathological, and genomic characterization of novel goose reovirus associated with liver necrosis in geese, China.
Since 2021, a novel strain of goose reovirus (GRV) has emerged within the goose farming industry in Guangdong province, China. This particular viral variant is distinguished by the presence of white necrotic foci primarily localized in the liver and spleen, leading to substantial economic losses for the poultry industry. However, the etiology, prevalence and genomic characteristics of the causative agent have not been thoroughly investigated. In this study, we conducted an epidemiological inquiry employing suspected GRV samples collected from May 2021 to September 2022. The macroscopic pathological and histopathological lesions associated with GRV-infected clinical specimens were examined. Moreover, we successfully isolated the GRV strain and elucidated the complete genome sequence of the isolate GD21/88. Through phylogenetic and recombination analysis, we unveiled that the GRV strains represent a novel variant resulting from multiple reassortment events. Specifically, the μNS, λC, and σNS genes of GRV were found to have originated from chicken reovirus, while the σA gene of GRV exhibited a higher degree of similarity with a novel duck reovirus. The remaining genes of GRV were traced back to Muscovy duck reovirus. Collectively, our findings underscore the significance of GRV as a pathogenic agent impacting the goose farming industry. The insights gleaned from this study contribute to a more comprehensive understanding of the epidemiology of GRV in Southern China and shed light on the genetic reassortment events exhibited by the virus.


Prevalence and phenotypic characterization of carbapenem-resistant Klebsiella pneumoniae strains recovered from sputum and fecal samples of ICU patients in Zhejiang Province, China.
To understand the prevalence and transmission of carbapenem-resistant Klebsiella pneumoniae (CRKP) in ICU patients in Zhejiang Province, China, and determined the genetic and phenotypic characteristics of these CRKP strains. A total of 202 ICU patients from eight tertiary hospitals were recruited and 55 non-duplicate CRKP strains were collected during July and August in 2017. These strains were subjected to determination of MICs, carriage of carbapenemase genes and tet(A) variants, PFGE, MLST and virulence potential using G. mellonella larvae infection model. A total of 55 CRKP strains were recovered from 42 patients, representing a carriage rate of 20.8%. CRKP strains were recovered from both the intestinal and respiratory tract of 13 patients. Importantly, strains isolated from sputum and fecal samples often displayed identical PFGE profiles, suggesting that CRKP may also colonize the respiratory tract. The most dominant ST type of these CRKP strains was ST11, accounting for 78% (43/55) of the test strains. The majority of CRKP strains were resistant to multiple antibiotics, with the exception of tigecycline and ceftazidime/avibactam. Interestingly, 32 strains were found to harbor the tet(A) variant, which is known to confer reduced tigecycline susceptibility. Assessment of the virulence potential of these CRKP strains by string test showed that results were negative for 53 of the 55 test strains. However, further assessment of virulence potential using a G. mellonella larvae infection model showed that CRKP isolated from sputum consistently exhibited a higher virulence level than strains recovered from fecal samples. CRKP is highly prevalent in ICU patients in Zhejiang Province with strains isolated from respiratory exhibiting higher virulence potential than those from GI tract. These data provide essential insight into development of new infection control measures to halt the transmission of CRKP in clinical settings.


Intracellular pH distribution as a cell health indicator in Saccharomyces cerevisiae.
Internal pH regulation is vital for many cell functions, including transport mechanisms and metabolic enzyme activity. More specifically, transport mechanisms are to a wide degree governed by internal pH distributions. We introduce the term standard deviation of the intracellular pH (s.d.(pH(int))) to describe the internal pH distributions. The cellular pH distributional response to external stress such as heat has not previously been determined. In this study, the intracellular pH (pH(i)) and the s.d.(pH(int)) of Saccharomyces cerevisiae cells exposed to supralethal temperatures were measured using fluorescence ratio imaging microscopy (FRIM). An exponential decline in pH(i) was observed after an initial small decline. For the first time, we report the use of FRIM for determining in vivo plasma membrane proton permeability coefficients in yeast. Furthermore, the exponential decay of pH(i) and the rupture of the cell plasma membrane, as measured by propidium iodide staining, at 70°C were not simultaneous but were separated by a significant temporal difference. Finally, a nonlinear relationship between the pH(i) and s.d.(pH(int)) was found; i.e. the s.d.(pH(int)) was significantly more sensitive to supralethal temperatures than pH(i). s.d.(pH(int)) is therefore proposed as an early health/vitality indicator in S. cerevisiae cells exposed to heat stress.


Costs and effectiveness of ultrasonography and limited computed tomography for diagnosing appendicitis in children.
A protocol of ultrasonography (US) followed by computed tomography with rectal contrast (CTRC) has been shown to be 94% accurate in the diagnosis of acute appendicitis in children. To evaluate the changes in patient management and costs of a protocol using US and CTRC in the evaluation of appendicitis in children. Prospective cohort study of 139 children between 3 and 21 years of age who had equivocal clinical findings for acute appendicitis seen in the emergency department of a large, urban pediatric teaching hospital between July 1998 and December 1998. Children with equivocal clinical presentations for acute appendicitis were prospectively evaluated with US. Patients with positive findings for acute appendicitis went directly to the operating room. Patients with negative or equivocal findings on US underwent CTRC. Surgical management plans were recorded before imaging, after US, and after CTRC. Surgical management plans before and after the imaging protocol as well as total hospital direct and indirect costs incurred or saved by each change in management were determined. Costs were obtained through the hospital's cost database and by ratios of costs to charges. Of the 139 children, the protocol resulted in a beneficial change in management in 86 children (61.9%), no change in management in 50 children (36.0%) and an incorrect change in management in 3 children (2.1%). US alone resulted in a beneficial change in management decision in 12/31 children (38.7%), while US followed by CTRC resulted in a beneficial change in management in 74/108 children (68.5%). The protocol resulted in a total cost savings of $78 503.99 or $565/patient. A protocol of US followed by CTRC in children with negative or equivocal US examinations results in a high rate of beneficial change in management as well as in total cost savings in children with equivocal clinical presentations for suspected appendicitis.


Why and when social support predicts older adults' pain-related disability: a longitudinal study.
Pain-related social support has been shown to be directly associated with pain-related disability, depending on whether it promotes functional autonomy or dependence. However, previous studies mostly relied on cross-sectional methods, precluding conclusions on the temporal relationship between pain-related social support and disability. Also, research on the behavioral and psychological processes that account for such a relationship is scarce. Therefore, this study aimed at investigating the following longitudinally: (1) direct effects of social support for functional autonomy/dependence on pain-related disability, (2) mediating role of physical functioning, pain-related self-efficacy, and fear, and (3) whether pain duration and pain intensity moderate such mediating processes. A total of 168 older adults (Mage = 78.3; SDage = 8.7) participated in a 3-month prospective design, with 3 moments of measurement, with a 6-week lag between them. Participants completed the Formal Social Support for Autonomy and Dependence in Pain Inventory, the Brief Pain Inventory, the 36-SF Health Survey, behavioral tasks from the Senior Fitness Test, the Pain Self-Efficacy Questionnaire, and the Tampa Scale for Kinesiophobia. Moderated mediation analyses showed that formal social support for functional dependence (T1) predicted an increase in pain-related disability (T3), that was mediated by self-reported physical functioning (T2) and by pain-related self-efficacy (T2) at short to moderate pain duration and at low to moderate pain intensity, but not at higher levels. Findings emphasized that social support for functional dependence is a risk factor for pain-related disability and uncovered the "why" and "when" of this relationship. Implications for the design of social support interventions aiming at promoting older adults' healthy aging despite chronic pain are drawn.


Corneal epithelial basement membrane changes in diabetes.
The corneas of 12 eye donors with maturity-onset diabetes were obtained, and the corneal epithelial basement membranes were studied by transmission electron microscopy. Similar tissue was obtained from 12 nondiabetic eye donors who were matched for age (within 2 years) and race. The mean thickness of the corneal epithelial basement membrane in nondiabetic patients was 0.33 micrometer (+/- 0.11 S.D.), which gives a normal range of 0.11 to 0.55 micrometer. None of the nondiabetic basement membranes lay outside this range. The basement membranes of four of the 12 diabetics exceeded this thickness. No race or sex difference was seen in basement membrane thickness, nor was a clear trend seen with age. Multilaminated basement membranes were seen in eight diabetic patients and six nondiabetic patients. Multilamination was more clearly related to basement membrane thickness than to the presence or absence of diabetes.


Isolation and partial characterization of a 30 kDa adhesin from Paracoccidioides brasiliensis.
The virulence of Paracoccidioides brasiliensis can be attenuated or lost after long periods of repeated subculturing and reestablished after animal inoculation. Only one adhesin (gp43) has been described until now, among the various identified components of P. brasiliensis, and gp43 shows adhesion to laminin. Thus, the present study was designed to isolate and characterize factors putatively related to the capacity of this fungus to adhere to the host by comparing P. brasiliensis samples, taken before and after animal inoculation. The two samples differed in their pattern of adhesion and invasion. The sample recently isolated from animals (Pb18b) demonstrated a greater capacity to adhere and to invade the Vero cells than the one subcultured in vitro (Pb18a). Extract from Pb18b also showed higher levels of protein expression than that from Pb18a, when two-dimensional electrophoresis gels were compared. A protein species of 30 kDa, pI 4.9, was more evident in the Pb18b extract and had properties of adhesin. Laminin, but none of the other extracellular matrix (ECM) components, such as fibronectin, collagen I and IV, bound specifically to the P. brasiliensis 30 kDa protein. The roles of 30 kDa and gp43 in cellular interactions were investigated and the adhesion of P. brasiliensis yeast cells was intensively inhibited by pre-treatment of epithelial cells with 30 kDa protein and gp43. Thus, this study presents evidence that adhesion capacity could be related to virulence, and that a 30 kDa adhesin accumulated differentially in samples with different levels of pathogenicity. This protein and its adhesion characteristics are being published for the first time and may be related to the virulence of P. brasiliensis.


Fluorescent single-stranded DNA binding protein as a probe for sensitive, real-time assays of helicase activity.
The formation and maintenance of single-stranded DNA (ssDNA) are essential parts of many processes involving DNA. For example, strand separation of double-stranded DNA (dsDNA) is catalyzed by helicases, and this exposure of the bases on the DNA allows further processing, such as replication, recombination, or repair. Assays of helicase activity and probes for their mechanism are essential for understanding related biological processes. Here we describe the development and use of a fluorescent probe to measure ssDNA formation specifically and in real time, with high sensitivity and time resolution. The reagentless biosensor is based on the ssDNA binding protein (SSB) from Escherichia coli, labeled at a specific site with a coumarin fluorophore. Its use in the study of DNA manipulations involving ssDNA intermediates is demonstrated in assays for DNA unwinding, catalyzed by DNA helicases.


Cell swelling-induced insulin secretion from INS-1E cells is inhibited by extracellular Ca2+ and is tetanus toxin resistant.
Cell swelling-induced insulin secretion represents an alternative pathway of stimulation of insulin secretion. INS-1E rat tumor beta cells do not release insulin in response to cell swelling in presence of Ca(2+) despite a good response to glucose challenge and appropriate increase in cell volume. Surprisingly, perifusion with Ca(2+)-depleted medium showed distinct secretory response of INS-1E cells to hypotonicity. Objective of this study was further characterization of the role of Ca(2+) in secretory process in INS-1 and INS-1E cell lines. Ca(2+) depleted hypotonic medium with 10 muM BAPTA/AM (intracellular chelator) induced insulin secretion from both types of cells. We demonstrated expression of L-type Ca(2+) channel Ca(v)1.2 and non-L-type Ca(2+) channels Ca(v)2.1 (P/Q-type), Ca(v)2.2 (N-type), and Ca(v)3.1 (T-type) in both cell lines. Inhibition of L type channel with nifedipine and/or P/Q type with omega-agatoxin IVA enabled distinct response to hypotonic medium also in INS-1E cells. Tetanus toxin (TeTx) in medium containing Ca(2+) and a group of calcium channel blockers inhibited hypotonicity-induced insulin secretion from INS-1 cells but not from INS-1E cells. Hypotonicity-induced insulin secretion from INS-1E cells is inhibited by extracellular Ca(2+), does not require intracellular Ca(2+) and is TeTx resistant.


Unexpected Roles of Interstitially Doped Lithium in Blue and Green Light Emitting Y2O3:Bi3+: A Combined Experimental and Computational Study.
To enhance the photoluminescence of lanthanide oxide, a clear understanding of its defect chemistry is necessary. In particular, when yttrium oxide, a widely used phosphor, undergoes doping, several of its atomic structures may be coupled with point defects that are difficult to understand through experimental results alone. Here, we report the strong enhancement of the photoluminescence (PL) of Y2O3:Bi3+ via codoping with Li+ ions and suggest a plausible mechanism for that enhancement using both experimental and computational studies. The codoping of Li+ ions into the Y2O3:Bi3+ phosphor was found to cause significant changes in its structural and optical properties. Interestingly, unlike previous reports on Li+ codoping with several other phosphors, we found that Li+ ions preferentially occupy interstitial sites of the Y2O3:Bi3+ phosphor. Computational insights based on density functional theory calculations also indicate that Li+ is energetically more stable in the interstitial sites than in the substitutional sites. In addition, interstitially doped Li+ was found to favor the vicinity of Bi3+ by an energy difference of 0.40 eV in comparison to isolated sites. The calculated DOS showed the formation of a shallow level directly above the unoccupied 6p orbital of Bi3+ as the result of interstitial Li+ doping, which may be responsible for the enhanced PL. Although the crystallinity of the host materials increased with the addition of Li salts, the degree of increase was minimal when the Li+ content was low (<1 mol %) where major PL enhancement was observed. Therefore, we reason that the enhanced PL mainly results from the shallow levels created by the interstitial Li+.


Mechanism analysis of a novel natural cationic modified dextran flocculant and its application in the treatment of blue algal blooms.
Blue algae, a type of harmful microalgae, are responsible for causing harmful algal blooms that result in severe environmental issues. To address this problem, a biopolysaccharide-based flocculant was developed for treating blue algae blooms. This flocculant was created by modifying high molecular weight dextran using the natural cationic monomer betaine (Dex-Bet), making it environmentally friendly. Various techniques were used to characterize the prepared Dex-Bet flocculant, including infrared spectroscopy (FTIR), nuclear magnetic resonance hydrogen spectroscopy (1H NMR), X-ray diffraction spectroscopy (XRD), field emission scanning electron microscopy (FESEM), and thermogravimetric analysis (TGA). The effectiveness of the Dex-Bet flocculant was evaluated using kaolin-simulated wastewater. The results showed that the treated supernatant had a transmittance of up to 98.25 %. Zeta potential analysis revealed that the main mechanisms of flocculation were charge neutralization, charge patching, and adsorption bridging. The application of Dex-Bet in treating blue-green algae resulted in a maximum removal rate of 98.2 %. This study provides a potential flocculant for blue algae bloom treatment.


[Reduction of the drowning risk for young children, but increased risk for children of recently immigrated non-Westerners].
To describe differences in the risk of drowning of young children (under 10 years) in the Netherlands according to ethnicity and relevant trends since 996. Retrospective. We analysed the causes of death data for all 266 children aged 0 to 10 years who died of drowning between 1996 and 2005. Information for the cause of death was obtained from the cause of death data of Statistics Netherlands. Data about the size and composition of the population at risk (age, sex and ethnicity) were obtained from the municipal population registers. Young children's risk of drowning has decreased by about one-third since 1996. This decrease took place among native Dutch children and children of the major ethnic groups, notably Turkish, Moroccan and Surinamese. However for children of recently immigrated parents of non-Western ethnicity, mainly asylum seekers, the risk of drowning was 4 to 8 times higher than that of native children aged 3 to 10 years. Timely health education directed at newly arrived families with children could be an important measure to help them cope with the hazards of living in a water-rich environment such as the Netherlands. The education should point out the necessity of increased supervision of the youngest children and improved swimming skills for the slightly older ones.


The ubiquitin-proteasome system in neurodegeneration.
Impairment of the ubiquitin-proteasome system (UPS) has been implicated in the pathogenesis of a wide variety of neurodegenerative disorders, including Alzheimer's, Parkinson's, and Huntington's diseases. The most significant risk factor for the development of these disorders is aging, which is associated with a progressive decline in UPS activity and the accumulation of oxidatively modified proteins. To date, no therapies have been developed that can specifically up-regulate this system. In the neurodegenerative brain, dysfunction of the UPS has been associated with the deposition of ubiquitinated protein aggregates and widespread disruption of the proteostasis network. Recent research has identified further evidence of impairment in substrate ubiquitination and proteasomal degradation, which could contribute to the loss of cellular proteostasis in neurodegenerative disease. Novel strategies for activation of the UPS by genetic manipulation and treatment with synthetic compounds have also recently been identified. Here, we discuss the specific roles of the UPS in the healthy central nervous system and establish how dysfunctional components can contribute to neurotoxicity in the context of disease. Knowledge of the UPS components that are specifically or preferentially involved in neurodegenerative disease will be critical in the development of targeted therapies which aim at limiting the accumulation of misfolded proteins without gross disturbance of this major proteolytic pathway.


Kinetics and mechanism of the general-acid-catalyzed ring-closure of the malondialdehyde-DNA adduct, N2-(3-oxo-1-propenyl)deoxyguanosine (N2OPdG-), to 3-(2'-Deoxy-beta-D-erythro-pentofuranosyl)pyrimido[1,2-alpha]purin- 10(3H)-one (M1dG).
3-(2'-Deoxy-beta-D-erythro-pentofuranosyl)pyrimido[1,2-alpha]purin-10(3H)-one (M1dG) is the major product of the reaction of deoxyguanosine with malondialdehyde (MDA). M1dG blocks replication by DNA polymerases in vitro and is mutagenic in vivo. M1dG reacts with hydroxide to form the N2-(3-oxo-1-propenyl)deoxyguanosine anion (N2OPdG-). This reaction is pH-dependent and reverses under neutral and acidic conditions to form M1dG. Here we describe the kinetics and mechanism of the ring-closure reaction in both the nucleoside and oligonucleotides. Kinetic analysis of absorbance and fluorescence changes demonstrates that ring-closure is biphasic, leading to the rapid formation of an intermediate that slowly converts to M1dG in a general-acid-catalyzed reaction. The dependence of the rate of the rapid phase on pH reveals the pKa for protonated N2OPdG is 6.9. One-dimensional 1H NMR and DQF-COSY experiments identified two distinct intermediates, N2OPdG-H and 8-hydroxy-6,7-propenodeoxyguanosine (HO-Prene-dG), that are formed upon acidification of N2OPdG-. Characterization of ring-closure in single-stranded and in melted duplex oligonucleotides shows M1dG formation is also acid-catalyzed in single-stranded oligonucleotides and that the denaturation of an oligonucleotide duplex enhances ring-closure. This work details the complexity of ring-closure in the nucleoside and oligonucleotides and provides new insight into the role of duplex DNA in catalyzing ring-opening and ring-closing of M1dG and N2OPdG.


Complementopathies and precision medicine.
The renaissance of complement diagnostics and therapeutics has introduced precision medicine into a widened field of complement-mediated diseases. In particular, complement-mediated diseases (or complementopathies) with ongoing or published clinical trials of complement inhibitors include paroxysmal nocturnal hemoglobinuria, cold agglutinin disease, hemolytic uremic syndrome, nephropathies, HELLP syndrome, transplant-associated thrombotic microangiopathy, antiphospholipid antibody syndrome, myasthenia gravis, and neuromyelitis optica. Recognizing that this field is rapidly expanding, we aim to provide a state-of-the-art review of (a) current understanding of complement biology for the clinician, (b) novel insights into complement with potential applicability to clinical practice, (c) complement in disease across various disciplines (hematology, nephrology, obstetrics, transplantation, rheumatology, and neurology), and (d) the potential future of precision medicine. Better understanding of complement diagnostics and therapeutics will not only facilitate physicians treating patients in clinical practice but also provide the basis for future research toward precision medicine in this field.


Male-biased dispersal in a fungus-gardening ant symbiosis.
For nearly all organisms, dispersal is a fundamental life-history trait that can shape their ecology and evolution. Variation in dispersal capabilities within a species exists and can influence population genetic structure and ecological interactions. In fungus-gardening (attine) ants, co-dispersal of ants and mutualistic fungi is crucial to the success of this obligate symbiosis. Female-biased dispersal (and gene flow) may be favored in attines because virgin queens carry the responsibility of dispersing the fungi, but a paucity of research has made this conclusion difficult. Here, we investigate dispersal of the fungus-gardening ant Trachymyrmex septentrionalis using a combination of maternally (mitochondrial DNA) and biparentally inherited (microsatellites) markers. We found three distinct, spatially isolated mitochondrial DNA haplotypes; two were found in the Florida panhandle and the other in the Florida peninsula. In contrast, biparental markers illustrated significant gene flow across this region and minimal spatial structure. The differential patterns uncovered from mitochondrial DNA and microsatellite markers suggest that most long-distance ant dispersal is male-biased and that females (and concomitantly the fungus) have more limited dispersal capabilities. Consequently, the limited female dispersal is likely an important bottleneck for the fungal symbiont. This bottleneck could slow fungal genetic diversification, which has significant implications for both ant hosts and fungal symbionts regarding population genetics, species distributions, adaptive responses to environmental change, and coevolutionary patterns.


[Influence of titanium dioxide on growth of Azotobacter vinelandii IMV V-7076].
Adding highly dispersed titanium dioxide particles of the culture medium has a stimulatory effect on growth of Azotobacter vinelandii. The maximum effect was observed when using 5 g/l and 10 g/l concentrations of this component. Under these conditions the number of grown bacteria is several times more than in the control variants. It was shown that the stimulatory effect could not be the consequence of saccharose sorption on the surface of particles of the dispersed compound. Possibly, the contact interaction of bacterial cell with titanium dioxide causes an increase of cell wall permeability which leads to an increase of substrates transport to the cell.


Stabilization of amphipathic alpha-helical and beta-helical conformations in synthetic peptides in the presence and absence of ionic interactions.
The role of ionic interactions in stabilizing amphipathic alpha-helices was studied in the synthetic peptide Ac-NLEELKKKLEELKG-NH2 (NLEKG14), potentially stabilized by attraction between complementary ions in successive turns of the helix, and in the peptide Ac-NLEELEEELEELEG-NH2 (NLEG14), in which no side-chain ionic attractions are possible. At a pH below the pKa of glutamate, NLEG14 had a higher helix content than NLEKG14. At pH 3 to pH 10, the helicity of NLEKG14 did not change, whereas NLEG14 was converted to random coil at pH 7. The role of ionic interactions in stabilizing the conformation of beta-structures was studied in the synthetic peptides Ac-KLKLKLELELELG-NH2 (KLEG13) and Ac-ELELELELELELG-NH2 (ELG13). At a pH below the pKa of glutamate, ELG13 had a higher beta-content than KLEG13, as judged by their dichroic spectra, but at higher pH, ELG13 was converted to random coil, whereas KLEG13 retained a predominantly beta-conformation. At pH 7, high NaCl concentration produced a significant increase in the alpha-helix content of NLEKG14, converted NLEG14 from random coil to alpha-helix and converted ELG13 from random coil to beta-conformation. Overall, the results demonstrate that ionic attraction between side-chains plays a lesser role than hydrogen bonding and hydrophobic effects in stabilizing the alpha- and beta-conformations exhibited by these amphipathic peptides.


RNA degradation in Saccharomyces cerevisae.
All RNA species in yeast cells are subject to turnover. Work over the past 20 years has defined degradation mechanisms for messenger RNAs, transfer RNAs, ribosomal RNAs, and noncoding RNAs. In addition, numerous quality control mechanisms that target aberrant RNAs have been identified. Generally, each decay mechanism contains factors that funnel RNA substrates to abundant exo- and/or endonucleases. Key issues for future work include determining the mechanisms that control the specificity of RNA degradation and how RNA degradation processes interact with translation, RNA transport, and other cellular processes.


Cardiovascular Outcome Trials in Type 2 Diabetes: What Do They Mean for Clinical Practice?
IN BRIEF Cardiovascular disease is the leading cause of morbidity and mortality in people with diabetes, and deaths from heart disease are two to four times higher among adults with type 2 diabetes. Trials such as the U.K. Prospective Diabetes Study, ACCORD (Action to Control Cardiovascular Risk in Diabetes), ADVANCE (Action in Diabetes and Vascular Disease: Preterax and Diamicron MR Controlled Evaluation), and VADT (Veteran's Affairs Diabetes Trial) produced mixed findings regarding whether intensive glycemic control results in improved cardiovascular (CV) outcomes for patients with diabetes. In response to concerns, including the CV safety of the thiazolidinedione rosiglitazone, the U.S. Food and Drug Administration and subsequently the European Medicines Agency issued guidance that trials should be conducted to prove that antihyperglycemic agents have acceptable CV risk profiles. In this article, the authors review the study designs and results of CV outcomes trials conducted with sodium-glucose cotransporter 2 inhibitors and glucagon-like peptide 1 receptor agonists and discuss how these may affect clinical practice.


The Ross procedure in children: preoperative haemodynamic manifestation has significant effect on late autograft re-operation.
The Ross procedure is the aortic valve-replacement procedure of choice in children. Nonetheless, late autograft re-operation for dilatation and/or valve regurgitation is of concern. We examined whether preoperative haemodynamic manifestation (e.g., stenosis, regurgitation and mixed aortic valve disease) affected late re-operation risk. Medical records of 227 children who underwent the Ross procedure (1991-2004) were reviewed. Competing-risks methodology determined time-related prevalence and associated factors for two mutually exclusive end-states after the Ross procedure: (1) death prior to subsequent autograft re-operation and (2) autograft re-operation, with the remainder of patients being alive and free from subsequent autograft re-operation. There were 162 male patients (71%) in this study. Median age at surgery was 12.1 years (range: 1 week-18 years). The haemodynamic aortic valve dysfunction was primarily stenosis (n=40, 18%), primarily regurgitation (n=109, 48%) and mixed disease (n=78, 35%). Underlying pathology was rheumatic fever (n=104, 46%), congenital heart disease (n=113, 50%) and endocarditis (n=8, 3%). Competing-risks analysis showed that, at 10 years following the Ross procedure, ∼5% of patients had died, 16% had undergone autograft re-operation with aortic valve replacement and 79% were alive and free from autograft re-operation. Ten-year freedom from autograft re-operation for patients with preoperative stenosis, regurgitation and mixed disease was 97%, 69% and 93%, respectively, (p<0.001 for regurgitation vs others). Risk factors for increased risk of autograft re-operation were rheumatic fever (parameter estimates (PEs): 2.09 ± 0.75, p=0.006), and earlier year of surgery (PE: 0.20 ± 0.06, p=0.001). Ten-year freedom from homograft replacement was 81% and was not dependent on haemodynamic manifestation (PE: -0.16 ± 0.38, p=0.68). Significant factors for homograft replacement included fresh homografts (PE: 2.2 ± 0.63, p=0.01) and annular enlargement (PE: 1.11 ± 0.3, p=0.01). Ten-year freedom from cardiac re-operation other than auto-/homograft was 85%, higher in patients with preoperative aortic regurgitation (PE: 1.01 ± 0.42, p=0.02). Concomitant cardiac surgery was a significant factor for late cardiac re-operation other than auto-/homograft replacement (PE: 1.79 ± 0.39, p<0.001). The Ross procedure in children is associated with excellent survival. Late autograft re-operation may be required; however, it is more common in children with preoperative aortic regurgitation, especially those with rheumatic fever. Better patient selection in later era has mitigated the risk of autograft re-operation. Continued improved candidate selection, along with modifications in autograft implantation and root/sinotubular stabilisation techniques, may further decrease late autograft failure.


A novel iron chelator in combination with a P-selectin antagonist prevents ischemia/reperfusion injury in a rat liver model.
Hepatic ischemia/reperfusion (I/R) injury is associated with early and late graft failure after liver transplantation. A major mechanism is leukocyte adhesion to endothelium followed by release of reactive oxygen intermediates. We examined whether desferriexochelin 772SM (D-Exo), a lipid soluble iron chelator that prevents hydroxyl radical formation, can enhance the capacity of recombinant P-selectin glycoprotein ligand immunoglobulin (rPSGL-Ig), a glycoprotein that binds to P-selectin and inhibits neutrophil adhesion, to protect against I/R injury in an ex vivo rat liver model. Rat livers were harvested and stored for 6 hr at 4 degrees C in University of Wisconsin solution and then perfused with oxygenated whole blood for 2 hr. Three groups were studied (n=6 rats/group): an untreated control group; a group that received 0.4 mg/kg rPSGL-Ig intraportally at the time of harvest; and a group that received 0.4 mg/kg rPSGL-Ig plus 1 micromol D-Exo intraportally both at the time of harvest and at the onset of reperfusion. Liver portal venous blood flow was assessed during perfusion, and at the end of each experiment, liver samples were collected for blinded histological evaluation and biochemical analyses. Livers treated with D-Exo + rPSGL-Ig had significantly higher blood flow than livers treated with rPSGL-1Ig alone (P<0.05), and both treatment groups had higher blood flow than controls (P<0.001). Production of carbonyl proteins, a protein oxidation product, was significantly reduced in the D-Exo + rPSGL-1Ig group (P<0.02 vs. controls), but not in the rPSGL-Ig alone group. Total reduced glutathione was significantly higher than controls in the D-Exo + rPSGL-Ig group (P<0.001 vs. controls), but not in the rPSGL-Ig alone group, indicating less oxidative stress in the D-Exo-treated group. Production of malondialdehyde, an index of lipid peroxidation, was significantly less than controls in both treatment groups (P<0.03). Histopathological findings paralleled these results with Banffs scores of 3.3+/-0.5, 1.8+/-0.4, and 1.3+/-0.5 in the control, rPSGL-Ig alone, and D-Exo plus rPSGL-Ig groups, resp. rPSGL-Ig provides partial protection against I/R injury to ex vivo rat livers; however, the addition of D-Exo substantially increases protection by reducing oxidative injury. These findings may have clinical relevance in preventing the consequences of I/R injury after liver transplantation.


Site-specific pausing of deoxyribonucleic acid synthesis catalyzed by four forms of Escherichia coli DNA polymerase III.
Sites on an fd DNA template which terminate synthesis catalyzed by each of four forms of Escherichia coli DNA polymerase III have been identified at single nucleotide resolution. Results were obtained by comparing the products made by forms of DNA polymerase III with products generated from the same 3'-terminus by using the dideoxynucleotide sequencing method, on high-resolution polyacrylamide gel electrophoresis. Each form of DNA polymerase III generates products of distinct lengths ending at a limited number of preferred sites of synthesis termination. The addition of auxiliary subunits to the DNA polymerase III core form of the enzyme has a distinct functional effect on primer elongation and specificity of polymerase pausing. Most sites (65%) can be correlated to positions of potential secondary structure in the template arising via local hydrogen-bonding interactions. The proximity of polymerase pausing to sites adjacent to hairpin stems was related to the size of the enzyme since the smaller core form of DNA polymerase III generally paused at sites which were closer to the base of these structures than the larger holoenzyme. The occurrence of termination sites is markedly affected by the inclusion of spermidine or Escherichia coli single-stranded DNA binding protein in the reaction mixtures. Additionally, a nucleotide composition specificity of pause sites has been observed.


Selective Targeting of Nanobody-Modified Gold Nanoparticles to Distinct Cell Types.
Nanobody-modified gold nanoparticles were used to explore their ability to achieve selective targeting in vitro and in vivo to distinct cell type(s), based on the specificity of the nanobody that was installed. We developed conjugation methods that exploit click chemistry for octahedral ∼50 nm gold nanoparticles and chiral ∼180 nm gold nanoparticles. We determined that each of these particles could be modified with ∼75 and ∼330 nanobodies, respectively. Particle-bound nanobodies retain their antigen binding capacity. After conjugation of the mouse Class II MHC-specific nanobody VHH7 to chiral gold nanoparticles, selective targeting of Class II MHC-positive cell types was observed in vitro by fluorometric assays and by dark-field microscopy. Upon installation of the positron emission tomography (PET) isotopes 89Zr or 64Cu on nanobody-modified gold nanoparticles and retro-orbital injection of the radiolabeled particles, we observed accumulation predominantly in the liver and to a far lesser extent in the spleen, regardless of the size of the gold nanoparticles and the identity of the attached nanobody. We observed a striking difference in the distribution of radioisotope-labeled gold nanoparticles by changing the route of administration to intraperitoneal delivery. Significantly reduced accumulation in the liver and spleen was observed by intraperitoneal injection of nanoparticles. In the case of nanobody-modified gold nanoparticles injected intraperitoneally, prominent and persistent signals from the parathymic lymph nodes were observed in the PET/computed tomography images.


Expression of prostate-specific membrane antigen in tumor-associated neovasculature of renal neoplasms.
Prostate-specific membrane antigen (PSMA) is highly expressed in prostate cancer cells. Recently, PSMA has been found in the neovasculature in association with other solid malignant tumors, including clear cell renal carcinoma (RCC). We studied the expression of PSMA in different primary renal tumors. A tissue microarray was constructed from 60 normal kidney, 21 clear cell RCC (CCRCC), 20 papillary RCC (PRCC), 16 chromophobe RCC, 19 oncocytoma, 14 transitional cell carcinoma, and 19 angiomyolipoma (AML) specimens. This tissue microarray was then immunostained for a vascular endothelial marker CD34 and PSMA. PSMA expression in CD34-positive tumor-associated neovasculature was scored according to the staining intensity and the percentage of vessels. Only diffuse strong or weak, or focal strong PSMA staining was graded as positive. PSMA was expressed in the proximal tubules of the normal kidney and in the tumor-associated vasculature in the renal tumors. Positive PSMA staining was detected in 76.2% of CCRCC, 31.2% of chromophobe RCC, 52.6% of oncocytoma, 21.4% of transitional cell carcinoma, and 0% of PRCC and AML specimens. Its expression was greater in CCRCC than PRCC, chromophobe RCC, transitional cell carcinoma, and AML (P <0.001), but was not significantly different from the expression in oncocytoma (P = 0.79). PSMA expression did not correlate with the pathologic stage in CCRCC. PSMA is differentially expressed in the tumor-associated neovasculature in different renal tumors. It is most commonly detected in CCRCC and rarely detectable in PRCC and AML. This finding suggests that antibodies against PSMA may potentially be used as a diagnostic marker and therapeutic target for renal neoplasms.


Assessment of regional lung recruitment and derecruitment during a PEEP trial based on electrical impedance tomography.
To investigate whether electrical impedance tomography (EIT) is capable of monitoring regional lung recruitment and lung collapse during a positive end-expiratory pressure (PEEP) trial. Experimental animal study of acute lung injury. Six pigs with saline-lavage-induced acute lung injury. An incremental and decremental PEEP trial at ten pressure levels was performed. Ventilatory, gas exchange, and hemodynamic parameters were automatically recorded. EIT and computed tomography (CT) scans of the same slice were simultaneously taken at each PEEP level. A significant correlation between EIT and CT analyses of end-expiratory gas volumes (r=0.98 up to 0.99) and tidal volumes (r=0.55 up to r=0.88) could be demonstrated. Changes in global and regional tidal volumes and arterial oxygenation (PaO2/FiO2) demonstrated recruitment/derecruitment during the trial, but at different onsets. During the decremental trial, derecruitment first occurred in dependent lung areas. This was indicated by lowered regional tidal volumes measured in this area and by a decrease of PaO2/FiO2. At the same time, the global tidal volume still continued to increase, because the increase of ventilation of the non-dependent areas was higher than the loss in the dependent areas. This indicates that opposing regional changes might cancel each other out when combined in a global parameter. EIT is suitable for monitoring the dynamic effects of PEEP variations on the regional change of tidal volume. It is superior to global ventilation parameters in assessing the beginning of alveolar recruitment and lung collapse.


A Dose-escalation Study of Recombinant Human Interleukin-18 in Combination With Ofatumumab After Autologous Peripheral Blood Stem Cell Transplantation for Lymphoma.
Interleukin-18 (IL-18) is an immunostimulatory cytokine that augments antibody-dependent cellular cytotoxicity mediated by human natural killer cells against antibody-coated lymphoma cells in vitro and that has antitumor activity in animal models. Ofatumumab is a CD20 monoclonal antibody with activity against human B-cell lymphomas. A phase I study of recombinant human (rh) IL-18 given with ofatumumab was undertaken in patients with CD20 lymphoma who had undergone high-dose chemotherapy and autologous peripheral blood stem cell transplantation. Cohorts of 3 patients were given intravenous infusions of ofatumumab 1000 mg weekly for 4 weeks with escalating doses of rhIL-18 as a intravenous infusion weekly for 8 consecutive weeks. Nine male patients with CD20 lymphomas were given ofatumumab in combination with rhIL-18 at doses of 3, 10, and 30 μg/kg. No unexpected or dose-limiting toxicities were observed. The mean reduction from predose levels in the number of peripheral blood natural killer cells after the first rhIL-18 infusion was 91%, 96%, and 97% for the 3, 10, and 30 μg/kg cohorts, respectively. Serum concentrations of interferon-γ and chemokines transiently increased following IL-18 dosing. rhIL-18 can be given in biologically active doses by weekly infusions in combination with ofatumumab after peripheral blood stem cell transplantation to patients with lymphoma. A maximum tolerated dose of rhIL-18 plus ofatumumab was not determined. Further studies of rhIL-18 and CD20 monoclonal antibodies in B-cell malignancies are warranted.


Evidence for Integrin - Venus Kinase Receptor 1 Alliance in the Ovary of Schistosoma mansoni Females Controlling Cell Survival.
In metazoan integrin signaling is an important process of mediating extracellular and intracellular communication processes. This can be achieved by cooperation of integrins with growth factor receptors (GFRs). Schistosoma mansoni is a helminth parasite inducing schistosomiasis, an infectious disease of worldwide significance for humans and animals. First studies on schistosome integrins revealed their role in reproductive processes, being involved in spermatogenesis and oogenesis. With respect to the roles of eggs for maintaining the parasite´s life cycle and for inducing the pathology of schistosomiasis, elucidating reproductive processes is of high importance. Here we studied the interaction of the integrin receptor Smβ-Int1 with the venus kinase receptor SmVKR1 in S. mansoni. To this end we cloned and characterized SmILK, SmPINCH, and SmNck2, three putative bridging molecules for their role in mediating Smβ-Int1/SmVKR1 cooperation. Phylogenetic analyses showed that these molecules form clusters that are specific for parasitic platyhelminths as it was shown for integrins before. Transcripts of all genes colocalized in the ovary. In Xenopus oocytes germinal vesicle breakdown (GVBD) was only induced if all members were simultaneously expressed. Coimmunoprecipitation results suggest that a Smβ-Int1-SmILK-SmPINCH-SmNck2-SmVKR1 complex can be formed leading to the phosphorylation and activation of SmVKR1. These results indicate that SmVKR1 can be activated in a ligand-independent manner by receptor-complex interaction. RNAi and inhibitor studies to knock-down SmILK as a representative complex member concurrently revealed effects on the extracellular matrix surrounding the ovary and oocyte localization within the ovary, oocyte survival, and egg production. By TUNEL assays, confocal laser scanning microscopy (CLSM), Caspase-3 assay, and transcript profiling of the pro-apoptotic BCL-2 family members BAK/BAX we obtained first evidence for roles of this signaling complex in mediating cell death in immature and primary oocytes. These results suggest that the Smβ-Int1/SmVKR1 signaling complex is important for differentiation and survival in oocytes of paired schistosomes.


Roles of nitric oxide synthases in platelet-activating factor-induced intestinal necrosis in rats.
To examine the role of constitutive and inducible nitric oxide synthases (cNOS and iNOS) in platelet-activating factor (PAF)-induced shock and intestinal injury. Prospective, randomized, controlled experimental study. Hospital research laboratory. Young adult male Sprague-Dawley rats were anesthetized and studied. Rats were injected with PAF, either alone or after the following pretreatments: a) selective iNOS inhibitors aminoguanidine or S-methylisothiourea; b) 3-morpholinosydnonimine, a NO donor; c) S-methylisothiourea + 3-morpholinosydnonimine; and d) antineutrophil antibody (to deplete neutrophils). Blood pressure, hematocrit, white blood cell counts, intestinal injury, and intestinal cNOS and iNOS activities were assessed. We found that: a) cNOS is the predominant NOS in the intestine and its activity is inversely correlated to the level of tissue injury; b) there is a time-dependent increase in cNOS activity in sham-operated animals, which was abolished by PAF; c) Western blotting and immunohistochemistry showed iNOS present in the normal intestine, localizing mainly in crypt cells; d) iNOS inhibitors attenuated PAF-induced injury in animals with high cNOS activity, but had no protective effect in animals with low cNOS activity; e) 3-morpholinosydnonimine, alone or together with S-methylisothiourea, alleviated PAF-induced injury; and f) neutrophil depletion blocked the suppressive effect of PAF on cNOS and prevented injury. We conclude that cNOS and iNOS play different roles in PAF-induced intestinal injury. Caution should be exerted concerning potential therapeutic uses of iNOS inhibitors.


Innervation of the trapezius muscle: a study in patients undergoing neck dissections.
Twenty-four patients with surgical section of the accessory nerve and/or its cervical contribution(s) were examined from 1 to 156 months after surgery, and compared to twenty controls. Thirteen patients had a classical neck dissection; seven had the whole length of the accessory nerve preserved but not the cervical plexus contributions. Four had the upper half of the accessory nerve sectioned, but with preservation of both the lower half and its cervical contributions. Clinical and electrophysiological studies of the three portions of the trapezius suggested the existence of an undescribed motor nerve supply to the trapezius and of a motor input from the cervical plexus contributions via the accessory nerve. The former is also supported by an anatomical study.


A new method for selection of angular aqueous plexus cells from porcine eyes: a model for Schlemm's canal endothelium.
The authors sought to develop a technique for isolating and culturing angular aqueous plexus (AAP) cells from more plentiful porcine eyes. AAP is an analogue of Schlemm's canal. Cells were differentially selected with puromycin, a toxin often used to select brain microvascular endothelial cells based on the expression of P-glycoprotein (P-gp), a multidrug resistance efflux pump. Trabecular meshwork containing AAP was dissected and pooled from fresh porcine eyes, digested in collagenase I, washed, filtered, and cultured for 8 days in a gelatin-coated plastic flask. Cells were then selected by exposure to 4 μg/mL puromycin for 2 days in the culture medium. Cells were fixed and immunostained for P-gp, ICAM II, von Willebrand factor (vWF), VE-cadherin, and α-smooth muscle actin (α-SMA). Histology of the limbus showed that the dissection was limited to the trabecular meshwork region, including the AAP. Before puromycin treatment, cells appeared heterogeneous and polygonal, suggestive of a mixed population. More than 90% of the cells were removed by puromycin, leaving a population that appeared uniformly cobblestone-like when grown to confluence and that was contact inhibited. Puromycin-selected cells stained positively for the endothelial markers ICAM II, vWF, and VE-cadherin but negatively for α-SMA, consistent with staining patterns in whole tissue. Based on marker expression, morphology, and behavior in culture, puromycin-selected cells from porcine outflow tissues are AAP endothelial cells. Thus, porcine eyes can provide a plentiful alternative cell source for studying Schlemm's canal biology related to ocular hypertension.


Stem cell factor is selectively secreted by arterial endothelial cells in bone marrow.
Endothelial cells (ECs) contribute to haematopoietic stem cell (HSC) maintenance in bone marrow, but the differential contributions of EC subtypes remain unknown, owing to the lack of methods to separate with high purity arterial endothelial cells (AECs) from sinusoidal endothelial cells (SECs). Here we show that the combination of podoplanin (PDPN) and Sca-1 expression distinguishes AECs (CD45- Ter119- Sca-1bright PDPN-) from SECs (CD45- Ter119- Sca-1dim PDPN+). PDPN can be substituted for antibodies against the adhesion molecules ICAM1 or E-selectin. Unexpectedly, prospective isolation reveals that AECs secrete nearly all detectable EC-derived stem cell factors (SCF). Genetic deletion of Scf in AECs, but not SECs, significantly reduced functional HSCs. Lineage-tracing analyses suggest that AECs and SECs self-regenerate independently after severe genotoxic insults, indicating the persistence of, and recovery from, radio-resistant pre-specified EC precursors. AEC-derived SCF also promotes HSC recovery after myeloablation. These results thus uncover heterogeneity in the contribution of ECs in stem cell niches.


Potential mechanisms underlying the cardiovascular benefits of sodium glucose cotransporter 2 inhibitors: a systematic review of data from preclinical studies.
There is growing evidence from Phase III randomized clinical trials of the cardiovascular benefits of sodium glucose cotransporter 2 (SGLT2) inhibitors in patients with diabetes mellitus. It is hypothesized that these benefits are mediated by mechanisms other than glucose control. To address this, we performed a systematic review of data from preclinical studies examining the direct cardioprotective effects of SGLT2 inhibitors. Medline, EMBASE, CINAHL, and International Pharmaceutical Abstracts databases were searched for preclinical studies that examined the potential cardioprotective effects of SGLT2 inhibitors. Submission documents to the US Food and Drug Administration, European Medicines Agency, and Japanese Pharmaceutical and Medical Devices Agency for the registration of SGLT2 inhibitors were also reviewed. A total of 36 reports were included in the final analysis. The potential direct cardiovascular benefits of SGLT2 inhibitors include: augmentation of signal transducer and activator of transcription 3; inhibition of sodium hydrogen exchange; reduction of atherosclerosis; modulation of natriuretic peptides; vasodilation; modulation of sympathetic tone; and reduction of inflammation, oxidative stress, endoplasmic reticulum stress, and cardiac glucose uptake via down-regulation of SGLT1 expression. There are a number of mechanisms by which SGLT2 inhibitors may exert cardiovascular benefits beyond glycaemic control.