Oleanolic acid administration alleviates neuropathic pain after a peripheral nerve injury by regulating microglia polarization-mediated neuroinflammation.
Neuropathic pain caused by a peripheral nerve injury constitutes a great challenge in clinical treatments due to the unsatisfactory efficacy of the current strategy. Microglial activation-mediated neuroinflammation is a major characteristic of neuropathic pain. Oleanolic acid is a natural triterpenoid in food and medical plants, and fulfills pleiotropic functions in inflammatory diseases. Nevertheless, its role in neuropathic pain remains poorly elucidated. In the current study, oleanolic acid dose-dependently suppressed LPS-evoked IBA-1 expression (a microglial marker) without cytotoxicity to microglia, suggesting the inhibitory efficacy of oleanolic acid in microglial activation. Moreover, oleanolic acid incubation offset LPS-induced increases in the iNOS transcript and NO releases from microglia, concomitant with the decreases in pro-inflammatory cytokine transcripts and production including IL-6, IL-1β, and TNF-α. Simultaneously, oleanolic acid shifted the microglial polarization from the M1 phenotype to the M2 phenotype upon LPS conditions by suppressing LPS-induced M1 marker CD16, CD86 transcripts, and enhancing the M2 marker Arg-1 mRNA and anti-inflammatory IL-10 levels. In addition, the LPS-induced activation of TLR4-NF-κB signaling was suppressed in the microglia after the oleanolic acid treatment. Restoring this signaling by the TLR4 plasmid transfection overturned the suppressive effects of oleanolic acid on microglial polarization-evoked inflammation. In vivo, oleanolic acid injection alleviated allodynia and hyperalgesia in SNL-induced neuropathic pain mice. Concomitantly, oleanolic acid facilitated microglial polarization to M2, accompanied by inhibition in inflammatory cytokine levels and activation of TLR4-NF-κB signaling. Collectively, these findings confirm that oleanolic acid may ameliorate neuropathic pain by promoting microglial polarization from pro-inflammatory M1 to anti-inflammatory M2 phenotype via the TLR4-NF-κB pathway, thereby indicating its usefulness as therapeutic intervention in neuropathic pain.


Type 1 parathyroid hormone receptor (PTH1R) nuclear trafficking: regulation of PTH1R nuclear-cytoplasmic shuttling by importin-alpha/beta and chromosomal region maintenance 1/exportin 1.
The type 1 PTH/PTH-related peptide receptor (PTH1R) is a class B G protein-coupled receptor that demonstrates immunoreactivity in the nucleus as well as cytoplasm of target cells. Our previous studies on the PTH1R have shown that it associates with the importin family of transport regulatory proteins. To investigate the role of the importins in PTH1R nuclear import, we used small interfering (si)RNA technology to knock down the expression of importin-beta in the mouse osteoblast-like cell line, MC3T3-E1. Immunofluorescence microscopy as well as ligand blotting for PTH1R in nuclear fractions of importin-beta siRNA-treated cells demonstrated a decrease in nuclear localization of the PTH1R in comparison with control cells. Under normal culture conditions, PTH1R is present in both the nucleus and cytoplasm of cells. Serum starvation favors nuclear localization of PTH1R, whereas returning cells to serum or treatment with PTH-related peptide induced its cytoplasmic localization. To address the nuclear export of PTH1R, interactions between PTH1R and chromosomal region maintenance 1 (CRM1) were investigated. PTH1R and CRM1 coimmunoprecipitated from MC3T3-E1 cells, suggesting that CRM1 and PTH1R form a complex in vivo. After treatment with leptomycin B, a specific inhibitor of CRM1-mediated nuclear export, PTH1R accumulated in the nucleus. Taken together, our studies show that PTH1R shuttles from the nucleus to the cytoplasm under normal physiological conditions and that this nuclear-cytoplasmic transport is dependent upon importin-alpha/beta and CRM1.


Quantitative changes in fissure sealant six months after placement.
Earlier studies have reported extensive volumetric loss of fissure sealant shortly after placement. To determine the volume of fissure sealant lost at six months, we used a servohydraulic profilometric apparatus in combination with computer graphics. Twenty-two premolar teeth were selected, and baseline silicone impressions were made of each tooth. The teeth were sealed and the occlusion adjusted where necessary. Impressions were made immediately of sealed teeth and again at six months. Epoxy replicas were made from all impressions, and their surfaces digitized by sequential profilometry. Using a three-dimensional root mean square (RMS) goodness-of-fit computer program, before sealant, after sealant, and six months after sealant computer-graphic images were superimposed. The volume of sealant lost after six months for all premolar teeth was X = 0.23 mm3 (maxillary second premolar: N = 9, X = 0.29 mm3; maxillary first premolar: N = 5, X = 0.27 mm3; mandibular second premolar: N = 4, X = 0.20 mm3; mandibular first premolar: N = 4, X = 0.08 mm3). The volume differences between tooth groups were not significant as measured by analysis of variance (ANOVA). These volumes represented a 13.99% loss of applied sealant for all premolars (maxillary second premolar: 19.72%; maxillary first premolar: 15.37%; mandibular second premolar: 8.46%; mandibular first premolar: 6.37%). The area of wear and the depth of wear also were measured.


MYC rearrangement but not extra MYC copies is an independent prognostic factor in patients with mantle cell lymphoma.
Mantle cell lymphoma (MCL) with MYC rearrangement (MYC-R) is rare and little is known about the importance of MYC extra copies (EC) in the absence of MYC-R in MCL patients. This study includes 88 MCL patients with MYC tested by fluorescence in situ hybridization and/or conventional cytogenetics, including 27 with MYC-R, 21 with MYC-EC, and 40 with normal (NL) MYC. MCL patients with MYC-R more often had blastoid/pleomorphic morphology; a higher frequency of CD10, MYC, and simultaneous MYC and BCL2 expression; a higher level of MYC; and a higher Ki67 proliferation rate (p<0.05) than those without MYC-R. Although patients with MYC-R more frequently received aggressive chemotherapy (p=0.001), their overall survival (OS) was significantly shorter than those without MYC-R. Compared with patients with MYC/BCL2 double hit lymphoma (DHL), patients with MYC-R MCL had a similar OS but more commonly had bone marrow involvement, stage 4 disease, and a different immunophenotype. MCL patients with MYC-EC showed an OS intermediate between those with MYC-R and MYC-NL, either all or only blastoid/pleomorphic MCL patients included. Multivariate analysis showed that MYC-R, but not MYC-EC, had an independent and negative impact on OS. In conclusion, MYC-R but not MYC-EC showed a higher MYC expression and is an adverse prognostic factor for MCL patients. Although the OS of MCL patients with MYC-R is similar to that of MYC/BCL2 DHL patients, these groups have different clinicopathologic features supporting the retention of MCL with MYC-R in the category of MCL, as recommended in the revised World Health Organization classification.


Relationship between serum levels of IL-18 and IgG1 in patients with primary Sjögren's syndrome, rheumatoid arthritis and healthy controls.
Primary Sjögren's syndrome (SS) is characterized by inflammation in salivary and lachrymal glands, with a local predominance of Th1-like cytokines, as well as the pleiotropic cytokine interleukin (IL) 18. High serum levels of polyclonal IgG are common, with a subclass imbalance in which IgG1 is increased and IgG2 is normal or low. IL-18 is also of pathogenetic importance in rheumatoid arthritis. In the present study we looked for any relationship between serum IL-18 as well as transforming growth factor (TGF) beta1 versus IgA, IgM, and IgG subclass levels in SS (n = 16), rheumatoid arthritis (RA) (n = 15), and healthy controls (n = 15). SS was defined by the revised American-European classification criteria. IL-18 and TGF-beta1 were analyzed with enzyme immunoassays (EIA), and IgG1, IgG2 and IgG3 by single radial immunodiffusion. In the composite group of RA, SS and normal controls, IgG1 and IL-18 were related (R = 0.52, P = 0.0005). No relation was found neither between IL-18 versus IgG2, IgG3 or IgA, nor between serum TGF-beta1 versus any of the immunoglobulins. Since serum levels of IL-18 are related to serum IgG1, IL-18 may be of importance for IgG1 switch and/or release.


Microdecompression for lumbar spinal canal stenosis.
A description of the technique for lumbar microdecompression and a prospective study of the outcomes. To describe and analyze a technique that affords an excellent decompression while minimizing damage to surrounding tissues. Commonly used techniques of lumbar decompression that include bilateral takedown of paraspinal musculature and aggressive bony resection can result in significant iatrogenic sequelae. A less destructive alternative is needed. Unilateral limited takedown of multifidus was undertaken, and ipsilateral decompression performed. The contralateral side then was addressed under the midline structures with microscopic visualization--thereby preserving the supra-/interspinous ligament complex and the contralateral musculature. Thirty consecutive patients undergoing the procedure were analyzed prospectively and after a follow-up period by independent observers using a modified validated functional outcome score and patient satisfaction measures. The technique affords an excellent decompression while minimizing destruction to tissues not directly involved in the pathologic process. Functional outcome scores doubled, and 87% of patients reported high satisfaction rates. Lumbar microdecompression is a minimally invasive technique that appears to provide excellent functional outcomes.


Biomechanical evaluation of intramedullary nail versus tension band fixation for transverse olecranon fractures.
To compare the biomechanical stability of an alternative custom-designed intramedullary nail (IMN) fixation with that of traditional tension band wiring (TBW), the gold standard for stabilizing transverse olecranon fractures. Ex vivo biomechanical study. Biomechanical laboratory. Six pairs of elderly cadaver elbows. A simulated transverse olecranon fracture was created in each elbow after it had been denuded of most of its soft tissue. The right and left olecranons of each pair were alternately assigned to the IMN or TBW group. A tensile force was applied via the triceps tendon while the elbow was secured in 90 degree of flexion. Displacement at the simulated fracture segment was measured optically. Fixation failure was defined as >2 mm of fracture displacement. The differences in stiffness and maximum load to failure between the two treatments were analyzed for significance (P < 0.05) using a one-tailed paired t test. IMN fixation was significantly stiffer and stronger than TBW fixation. The locked IMN provided stronger and stiffer fixation than did TBW. Theoretically the IMN fixation would require less surgical exposure and would be expected to require fewer revisions than TBW fixation. IMN fixation warrants consideration as a clinical alternative to TBW.


Nanoconfined polymerization limits crack propagation in hysteresis-free gels.
Consecutive mechanical loading cycles cause irreversible fatigue damage and residual strain in gels, affecting their service life and application scope. Hysteresis-free hydrogels within a limited deformation range have been created by various strategies. However, large deformation and high elasticity are inherently contradictory attributes. Here we present a nanoconfined polymerization strategy for producing tough and near-zero-hysteresis gels under a large range of deformations. Gels are prepared through in situ polymerization within nanochannels of covalent organic frameworks or molecular sieves. The nanochannel confinement and strong hydrogen bonding interactions with polymer segments are crucial for achieving rapid self-reinforcement. The rigid nanostructures relieve the stress concentration at the crack tips and prevent crack propagation, enhancing the ultimate fracture strain (17,580 ± 308%), toughness (87.7 ± 2.3 MJ m-3) and crack propagation strain (5,800%) of the gels. This approach provides a general strategy for synthesizing gels that overcome the traditional trade-offs of large deformation and high elasticity.


Basis for experimental chemotherapy in lung cancer.
The in vitro chemotherapy sensitivity of five well-characterized lung cancer cell lines (two small cell and three non-small cell) was evaluated with six single agents (mitomycin C, vincristine, cisplatin, thiotepa, vinblastine, and etoposide [VP-16]) and two combination regimens (cisplatin plus mitomycin C and cisplatin plus etoposide [VP-16]). The results correlated closely with the clinical responsiveness of these tumor types. The small cell lung cancer (SCLC) lines were generally more sensitive than the non-small cell lung cancer (NSCLC) lines to vincristine, thiotepa, and etoposide (VP-16). Vinblastine was significantly less active than vincristine in SCLC lines and more active than vincristine in NSCLC lines. The two combinations that were tested demonstrated additive cytotoxicity. The data support the use of these cell lines for the selection of new anticancer agents and for the development of new treatment strategies.


Are European Blue Economy ambitions in conflict with European environmental visions?
We report the outcomes of a comprehensive study of the potential consequences of the implementation of the EU Maritime Spatial Planning Directive (MSPD) in Danish waters. The analyses are anchored in a framework developed in support of data-driven Ecosystem-Based Maritime Spatial Planning. The data for the models include not only human stressors but also information on the distribution of ecosystem components ranging from planktonic communities over benthic communities to fish, seabirds and marine mammals. We have established a baseline, based on state-of-the-art data sets, with respect to combined effects upon ecosystem components. Future scenarios for the developments in human stressors were estimated for 2030 and 2050 based on information on existing policies, strategies and plans and were compared to the baseline. In addition, we developed a scenario for implementation of the Marine Strategy Framework Directive (MSFD), i.e. working towards meeting the objectives of Good Environmental Status. Our results indicate that (1) combined human stressors will possibly increase in 2030 and 2050 compared to the baseline, (2) increased combined human stressors are likely to lead to a worsening of the environmental and ecological status sensu the Marine Strategy Framework Directive and the Water Framework Directive (WFD), and (3) the MSPD implementation process appears to conflict with the MSFD and WFD objectives. Accordingly, we are sceptical of claims of an untapped potential for Blue Growth in Danish marine waters.


Improved elongation factor-1 alpha-based vectors for stable high-level expression of heterologous proteins in Chinese hamster ovary cells.
Establishing highly productive clonal cell lines with constant productivity over 2-3 months of continuous culture remains a tedious task requiring the screening of tens of thousands of clonal colonies. In addition, long-term cultivation of many candidate lines derived in the absence of drug selection pressure is necessary. Expression vectors based on the elongation factor-1 alpha (EEF1A) gene and the dihydrofolate reductase (DHFR) selection marker (with separate promoters) can be used to obtain highly productive populations of stably transfected cells in the selection medium, but they have not been tested for their ability to support target gene amplification under gradually increasing methotrexate pressure. We have modified EEF1A-based vectors by linking the DHFR selection marker to the target gene in the bicistronic RNA, shortening the overall plasmid size, and adding an Epstein-Barr virus terminal repeat fragment (EBVTR) element. Presence of the EBVTR element increased the rate of stable transfection by the plasmid by 24 times that of the EBVTR-minus control and improved the rate of methotrexate-driven gene amplification. The mean expression level of the enhanced green fluorescent protein (eGFP) used herein as a model protein, increased up to eight-fold using a single round of amplification in the case of adherent colonies formation and up to 4.5-fold in the case of suspension polyclonal cultures. Several eGFP-expressing cell populations produced using vectors with antibiotic resistance markers instead of the DHFR marker were compared with each other. Stable transfection of Chinese hamster ovary (CHO) DG44 cells by the p1.2-Hygro-eGFP plasmid (containing a hygromycin resistance marker) generated highest eGFP expression levels of up to 8.9% of the total cytoplasmic protein, with less than 5% of the cell population being eGFP-negative. The p1.1 vector was very effective for stable transfection of CHO cells and capable of rapid MTX-driven target gene amplification, while p1.2-Hygro achieved similar eGFP expression levels as p1.1. The set of vectors we have developed should speed-up the process of generating highly productive clonal cell lines while substantially decreasing the associated experimental effort.


Evaluation of Prehospital Blood Products to Attenuate Acute Coagulopathy of Trauma in a Model of Severe Injury and Shock in Anesthetized Pigs.
Acute trauma coagulopathy (ATC) is seen in 30% to 40% of severely injured casualties. Early use of blood products attenuates ATC, but the timing for optimal effect is unknown. Emergent clinical practice has started prehospital deployment of blood products (combined packed red blood cells and fresh frozen plasma [PRBCs:FFP], and alternatively PRBCs alone), but this is associated with significant logistical burden and some clinical risk. It is therefore imperative to establish whether prehospital use of blood products is likely to confer benefit. This study compared the potential impact of prehospital resuscitation with (PRBCs:FFP 1:1 ratio) versus PRBCs alone versus 0.9% saline (standard of care) in a model of severe injury. Twenty-four terminally anesthetised Large White pigs received controlled soft tissue injury and controlled hemorrhage (35% blood volume) followed by a 30-min shock phase. The animals were allocated randomly to one of three treatment groups during a 60-min prehospital evacuation phase: hypotensive resuscitation (target systolic arterial pressure 80 mmHg) using either 0.9% saline (group 1, n = 9), PRBCs:FFP (group 2, n = 9), or PRBCs alone (group 3, n = 6). Following this phase, an in-hospital phase involving resuscitation to a normotensive target (110 mmHg systolic arterial blood pressure) using PRBCs:FFP was performed in all groups. There was no mortality in any group. A coagulopathy developed in group 1 (significant increase in clot initiation and dynamics shown by TEG [thromboelastography] R and K times) that persisted for 60 to 90 min into the in-hospital phase. The coagulopathy was significantly attenuated in groups 2 and 3 (P = 0.025 R time and P = 0.035 K time), which were not significantly different from each other. Finally, the volumes of resuscitation fluid required was significantly greater in group 1 compared with groups 2 and 3 (P = 0.0067) (2.8 ± 0.3 vs. 1.9 ± 0.2 and 1.8 ± 0.3 L, respectively). This difference was principally due to a greater volume of saline used in group 1 (P = 0.001). Prehospital PRBCs:FFP or PRBCs alone may therefore attenuate ATC. Furthermore, the amount of crystalloid may be reduced with potential benefit of reducing the extravasation effect and later tissue edema.


A membrane-bound eIF2 alpha kinase located in endosomes is regulated by heme and controls differentiation and ROS levels in Trypanosoma cruzi.
Translation initiation has been described as a key step for the control of growth and differentiation of several protozoan parasites in response to environmental changes. This occurs by the activation of protein kinases that phosphorylate the alpha subunit of the translation initiation factor 2 (eIF2α), which decreases translation, and in higher eukaryotes favors the expression of stress remedial response genes. However, very little is known about the signals that activate eIF2α kinases in protozoan parasites. Here, we characterized an eIF2α kinase of Trypanosoma cruzi (TcK2), the agent of Chagas' disease, as a transmembrane protein located in organelles that accumulate nutrients in proliferating parasite forms. We found that heme binds specifically to the catalytic domain of the kinase, inhibiting its activity. In the absence of heme, TcK2 is activated, arresting cell growth and inducing differentiation of proliferative into infective and non-proliferative forms. Parasites lacking TcK2 lose this differentiation capacity and heme is not stored in reserve organelles, remaining in the cytosol. TcK2 null cells display growth deficiencies, accumulating hydrogen peroxide that drives the generation of reactive oxygen species. The augmented level of hydrogen peroxide occurs as a consequence of increased superoxide dismutase activity and decreased peroxide activity. These phenotypes could be reverted by the re-expression of the wild type but not of a TcK2 dead mutant. These findings indicate that heme is a key factor for the growth control and differentiation through regulation of an unusual type of eIF2α kinase in T. cruzi.


Reflecting on Progress in and Establishing Benchmarks for Sex and Gender Health Education.
Sex and gender influence every aspect of human health, thus, sex- and gender-related topics should be incorporated in all aspects of health education curricula. Sex and gender health education (SGHE) is the rigorous, intersectional, data-driven integration of sex and gender into all elements of health education. A multisectoral group of thought leaders has collaborated to advance SGHE since 2012. This cross-sector collaboration to advance SGHE has been successful on several fronts, primarily developing robust interprofessional SGHE programs, hosting a series of international SGHE summits, developing sex- and gender-specific resources, and broadening the collaboration beyond medical education. However, other deeply entrenched challenges have proven more difficult to address, including accurate and consistent sex and gender reporting in research publications, broadening institutional support for SGHE, and the development and implementation of evaluation plans for assessing learner outcomes and the downstream effects of SGHE on patient care. This commentary reflects on progress made in SGHE over the first decade of the current collaboration (2012-2022), articulates a vision for next steps to advance SGHE, and proposes 4 benchmarks to guide the next decade of SGHE: (1) integrate sex, gender, and intersectionality across health curricula; (2) develop sex- and gender-specific resources for health professionals; (3) improve sex and gender reporting in research publications; and (4) develop evaluation plans to assess learner and patient outcomes.


Transit time homogenization in ischemic stroke - A novel biomarker of penumbral microvascular failure?
Cerebral ischemia causes widespread capillary no-flow in animal studies. The extent of microvascular impairment in human stroke, however, is unclear. We examined how acute intra-voxel transit time characteristics and subsequent recanalization affect tissue outcome on follow-up MRI in a historic cohort of 126 acute ischemic stroke patients. Based on perfusion-weighted MRI data, we characterized voxel-wise transit times in terms of their mean transit time (MTT), standard deviation (capillary transit time heterogeneity - CTH), and the CTH:MTT ratio (relative transit time heterogeneity), which is expected to remain constant during changes in perfusion pressure in a microvasculature consisting of passive, compliant vessels. To aid data interpretation, we also developed a computational model that relates graded microvascular failure to changes in these parameters. In perfusion-diffusion mismatch tissue, prolonged mean transit time (>5 seconds) and very low cerebral blood flow (≤6 mL/100 mL/min) was associated with high risk of infarction, largely independent of recanalization status. In the remaining mismatch region, low relative transit time heterogeneity predicted subsequent infarction if recanalization was not achieved. Our model suggested that transit time homogenization represents capillary no-flow. Consistent with this notion, low relative transit time heterogeneity values were associated with lower cerebral blood volume. We speculate that low RTH may represent a novel biomarker of penumbral microvascular failure.


Visible-Light Photocatalytic Overall Water Splitting on a B4C3/CxNy Z-Scheme Heterojunction: Role of Ultrafast Carrier Recombination-Transfer Kinetics.
Herein, combining density functional theory (DFT) calculations with nonadiabatic molecular dynamics (NAMD), we built a computational framework to rationally screen from a series of 2D conjugated carbon nitrides (CNs) to match with B4C3, resulting in the excellent direct Z-scheme photocatalyst (B4C3/C6N6) for overall water splitting (OWS). Studies on interface engineering and ultrafast dynamics of carrier recombination-transfer show that in the B4C3/C6N6 system, compared with the slower interlayer migration process of carriers, strong nonadiabatic coupling and longer quantum decoherence time accelerates weak carrier interlayer recombination on a subpicosecond time scale, enabling simultaneous triggering of hydrogen evolution reaction (HER) with ΔG = -0.23 eV and spontaneous oxygen evolution reaction (OER) in the absence of sacrificial or cocatalysts. In general, our work will promote the design of efficient direct Z-scheme photocatalysts from an ultrafast dynamics perspective.


Occupational exposure to chemical agents in the particleboard industry.
Most of the measurements concerned formaldehyde and wood dust. The other substances measured included terpenes, solvents, and heptachlor. Before 1975 the formaldehyde concentration regularly exceeded 2 ppm during many workphases. Considerable improvements in ventilation and the composition of glues have occurred since then, and today the exposure level is below 2 ppm during most workphases. High peak concentrations, 20-30 ppm at the highest, were characteristic of exposure in earlier years. The concentrations of wood dust have also decreased, eg, from over 5 mg/m3 to 1 mg/m3 or below during forming. These data have been used to evaluate past exposures in an epidemiologic study on cancer risks in the particleboard, plywood, and sawmill industries.


Piperlongumine promotes autophagy via inhibition of Akt/mTOR signalling and mediates cancer cell death.
The Akt/mammalian target of rapamycin (mTOR) signalling pathway serves as a critical regulator of cellular growth, proliferation and survival. Akt aberrant activation has been implicated in carcinogenesis and anticancer therapy resistance. Piperlongumine (PL), a natural alkaloid present in the fruit of the Long pepper, is known to exhibit notable anticancer effects. Here we investigate the impact of PL on Akt/mTOR signalling. We examined Akt/mTOR signalling in cancer cells of various origins including prostate, kidney and breast after PL treatment. Furthermore, cell viability after concomitant treatment with PL and the autophagy inhibitor, Chloroquine (CQ) was assessed. We then examined the efficacy of in vivo combination treatment using a mouse xenograft tumour model. We demonstrate for the first time that PL effectively inhibits phosphorylation of Akt target proteins in all tested cells. Furthermore, the downregulation of Akt downstream signalling resulted in decrease of mTORC1 activity and autophagy stimulation. Using the autophagy inhibitor, CQ, the level of PL-induced cellular death was significantly increased. Moreover, concomitant treatment with PL and CQ demonstrated notable antitumour effect in a xenograft mouse model. Our data provide novel therapeutic opportunities to mediate cancer cellular death using PL. As such, PL may afford a novel paradigm for both prevention and treatment of malignancy.


Comparison of nutritional indices and outcome in critically ill patients.
Seventeen critically ill patients were studied prospectively to determine the relationship of serum albumin, creatinine-height index (CHI) and weight-height index (WHI) to clinical outcome. Use of the conventional "normal values" as cut-offs failed to discriminate between survivors and nonsurvivors better than chance alone. A serum albumin less than 2.5 g/dl correctly separated 93% of the patients in terms of survival prognosis.


Genomics of natural history collections for understanding evolution in the wild.
A long-standing question in biology is how organisms change through time and space in response to their environment. This knowledge is of particular relevance to predicting how organisms might respond to future environmental changes caused by human-induced global change. Usually researchers make inferences about past events based on an understanding of current static genetic patterns, but these are limited in their capacity to inform on underlying past processes. Natural history collections (NHCs) represent a unique and critical source of information to provide temporally deep and spatially broad time-series of samples. By using NHC samples, researchers can directly observe genetic changes over time and space and link those changes with specific ecological/evolutionary events. Until recently, such genetic studies were hindered by the intrinsic challenges of NHC samples (i.e. low yield of highly fragmented DNA). However, recent methodological and technological developments have revolutionized the possibilities in the novel field of NHC genomics. In this Special Feature, we compile a range of studies spanning from methodological aspects to particular case studies which demonstrate the enormous potential of NHC samples for accessing large genomic data sets from the past to advance our knowledge on how populations and species respond to global change at multiple spatial-temporal scales. We also highlight possible limitations, recommendations and a few opportunities for future researchers aiming to study NHC genomics.


Prevention of more complications in patients with head trauma.
Head trauma and brain injuries are common causes of emergency admission and usually predispose multiple psychiatric complications. In turn, the traumas often occur after some psychiatric disorders. Therefore, the complicated interaction of these factors often causes difficulties in diagnosis and management of the patients. The side-effects of surgical and medical treatments may also complicate these processes more, as well. In this study, we present a case of a young patient with these complex factors and discuss the diagnosis and management.


Brief telephone intervention increases testing for osteoporosis in patients treated in emergency departments for wrist fractures.
Previous studies show that identification and treatment of osteoporosis in patients with minimal trauma fractures treated as outpatients are poor. Our aim was to test two interventions designed to increase rates of identification and treatment. This prospective, action research study, using explicit medical record review and scripted telephone interview, was conducted at emergency departments (ED) of three hospitals from April 2007 to February 2008. Participants were patients aged over 50 years who were treated as outpatients with a minimal trauma wrist fracture. Data collected included demographic and fracture details, bone density testing and osteoporosis-related medication change. There were two interventions staff education in ED and fracture clinic and information provided to patients by telephone by a research nurse. These interventions were applied to all patients sequentially. The outcome measure of interest was the proportion of patients who underwent bone density testing (DEXA scans) in the follow-up period, analysed by intervention (clinic or phone). One hundred and seventeen patients were studied. Eighty-six per cent were female; median age 64 years. Ten per cent (12/117) of the ED/clinic intervention group had undergone testing at follow up. At follow up after the telephone intervention 55% (65/117) had undergone testing (P < 0.001, chi(2)). Patients undergoing testing were significantly more likely to have an osteoporosis-related medication change (relative risk 6.8, 95% CI 2.8-17.9). A brief telephone intervention and provision of information pack significantly improved testing rates for osteoporosis after minimal trauma wrist fracture. An ED/clinic-based intervention resulted in low rates of testing. Treatment of clinical osteoporosis remains suboptimal.


Fundamental research and application of the specific fluidic behavior of mixed solvents in a microspace.
The author herein reviews a specific microfluidic behavior exhibited by mixed-solvent solutions in a microspace, coined as the tube radial distribution phenomenon (TRDP). The specific fluidic behavior was observed in the following solution systems: ternary water-hydrophilic/hydrophobic organic solvents, water-surfactant, water-ionic liquid, and fluorous/organic solvents. When the mixed homogeneous solutions were delivered into a microspace under certain conditions, the solvent molecules were radially distributed in the microspace, generating inner and outer phases with a kinetic liquid-liquid interface. The TRDP was fundamentally evaluated by fluorescence microscopy, phase diagrams construction, and the elution behaviors of solutes in a capillary tube. A TRDP-based capillary chromatography, referred to as tube radial distribution chromatography (TRDC), where the outer phase serves as a pseudo-stationary phase under laminar flow conditions, has been developed as one of the applications of TRDP. We have also investigated TRDP-based extraction, chemical reaction, and mixing processes, coined as tube radial distribution extraction (TRDE), tube radial distribution reaction (TRDR), and tube radial distribution mixing (TRDM), respectively. The concept and experimental findings regarding TRDP, TRDC, TRDE, TRDR, and TRDM are described in this review.


pERK, pAKT and p53 as tissue biomarkers in erlotinib-treated patients with advanced pancreatic cancer: a translational subgroup analysis from AIO-PK0104.
The role of pERK, pAKT and p53 as biomarkers in patients with advanced pancreatic cancer has not yet been defined. Within the phase III study AIO-PK0104 281 patients with advanced pancreatic cancer received an erlotinib-based 1st-line regimen. Archival tissue from 153 patients was available for central immunohistochemistry staining for pERK, pAKT and p53. Within a subgroup analysis, biomarker data were correlated with efficacy endpoints and skin rash using a Cox regression model. Fifty-five out of 153 patients were classified as pERKlow and 98 patients as pERKhigh; median overall survival (OS) was 6.2 months and 5.7 months, respectively (HR 1.29, p = 0.16). When analysing pERK as continuous variable, the pERK score was significantly associated with OS (HR 1.06, 95% CI 1.0-1.12, p = 0.05). Twenty-one of 35 patients were pAKTlow and 14/35 pAKThigh with a corresponding median OS of 6.4 months and 6.8 months, respectively (HR 1.03, p = 0.93). Four out of 50 patients had a complete loss of p53 expression, 20 patients a regular expression and 26 patients had tumors with p53 overexpression. The p53 status had no impact on OS (p = 0.91); however, a significant improvement in progression-free survival (PFS) (6.0 vs 1.8 months, HR 0.24, p = 0.02) and a higher rate of skin rash (84% vs 25%, p = 0.02) was observed for patients with a regular p53 expression compared to patients with a complete loss of p53. pERK expression may have an impact on OS in erlotinib-treated patients with advanced pancreatic cancer; p53 should be further investigated for its potential role as a predictive marker for PFS and skin rash. NCT00440167 (registration date: February 22, 2007).


Characterization of trans-septal activation during septal pacing: criteria for identification of intramural ventricular tachycardia substrate in nonischemic cardiomyopathy.
Identification of intramural basal-septal ventricular tachycardia (VT) substrate is challenging in nonischemic cardiomyopathy. We sought to (1) characterize normal/abnormal trans-septal right ventricular (RV) to left ventricular activation; (2) assess the effect of opposite RV pacing on left ventricular septal bipolar electrograms (EGMs); and (3) establish criteria for the identification of intramural septal VT substrate. Endocardial activation mapping and local EGM assessment of the left interventricular septum was performed during RV basal septal pacing in 40 patients undergoing VT ablation with no evidence of septal scar (group 1, n=14) and with septal scar (group 2, n=26) defined by low septal unipolar voltage (<8.3 mV) and delayed enhancement on cardiac MRI with/without abnormal bipolar voltage (<1.5 mV) in sinus rhythm. Left ventricular trans-septal activation time was prolonged in Group 2 compared with Group 1 (55.3±33.0 versus 25.7±8.8 ms; P=0.003). In 6 group 2 patients, left ventricular septal breakthrough was displaced to the scar border. During RV pacing, group 2 had fractionated (8.8%), late (2.8%), and split (5.7%) EGMs not seen in group 1. Trans-septal activation >40 ms (sensitivity 60%, specificity 100%; P<0.001) and EGM duration >95 ms during pacing (sensitivity 22%, specificity 91%; P<0.001) identified septal scar (13/26 pts). In patients with nonischemic cardiomyopathy, VT and septal scar, delayed transmural conduction time (>40 ms) and fractionated, late, split, and wide (>95 ms) bipolar EGMs during RV basal pacing identify intramural VT substrate. In select cases, the basal septum appears compartmentalized as the stimulated wavefront is rerouted to the scar border.


Cost-Effectiveness Analysis of Fibrinolysis vs Thoracoscopic Decortication for Early Empyema.
Surgical decortication is recommended by national guidelines for management of early empyema, but intrapleural fibrinolysis is frequently used as a first-line therapy in clinical practice. This study compared the cost-effectiveness of video-assisted thoracoscopic surgery (VATS) decortication with intrapleural fibrinolysis for early empyema. A decision analysis model was developed. The base clinical case was a 65-year-old man with early empyema treated either by VATS decortication or intrapleural tissue plasminogen activator and deoxyribonuclease. The likelihood of key outcomes occurring was derived from the literature. Medicare diagnosis-related groups and manufacturers' drug prices were used for cost estimates. Successful treatment was defined as complete or nearly complete resolution of empyema on imaging. Effectiveness was defined as health utility 1 year after empyema. Intrapleural tissue plasminogen activator and deoxyribonuclease were more cost-effective than VATS decortication for treating early empyema for the base clinical case. Surgical decortication had a slightly lower cost than fibrinolysis ($13,345 vs $13,965), but fibrinolysis had marginally higher effectiveness at 1 year (health utility of 0.80 vs 0.71). Therefore, fibrinolysis was the more cost-effective option. Sensitivity analyses found that fibrinolysis as the initial therapy was more cost-effective when the probability of success was greater than 60% or the initial cost was less than $13,000. Surgical decortication and intrapleural fibrinolysis have nearly equivalent cost-effectiveness for early empyema in patients who can tolerate both procedures. Surgeons should consider patient-specific factors, as well as the cost and effectiveness of both modalities, when deciding on an initial treatment for early empyema.


The fate of bisphenol A, 4-tert-octylphenol and 4-nonylphenol leached from plastic debris into marine water--experimental studies on biodegradation and sorption on suspended particulate matter and nano-TiO2.
Experiments were carried out, the aim of which was to determine the leaching rates for bisphenol A (4,4'-(propane-2,2-diyl)diphenol - BPA), 4-tert-octylphenol (OP), 4-nonylphenol (NP) from polycarbonate and recycled tyre granules into the seawater. Additionally biodegradation, sorption on marine suspended particulate matter and sorption on various types of nano-TiO2 of BPA, OP, NP were studied. Experiments were carried out on plastics at various stages of degradation. The conducted experiment confirmed the flux of BPA, OP and NP from the studied plastics into seawater. The initial photodegradation of the plastic had a significant influence on the amount of the studied components released into water. During the first days of the experiment leaching was weaker from aged materials. After 60 days leaching of BPA and OP was higher for aged plastic compared to unaged. On average, suspension adsorbed OP, BPA and NP from seawater at respective levels of 37%, 75% and 100%. On the other hand, during biodegradation on average 25%, 9% and 2% of OP, BPA and NP respectively are removed from water. Nano-TiO2 of 21 nm pore size diameter adsorbed all the compounds more strongly than nano-TiO2 of 15 nm pores sized coated with Al and stearic acid. The strongest sorption (100%) on different types of nano-TiO2 was that of the most hydrophobic and more linear structured NP with just one phenol group. The weakest sorption was observed in the case of BPA, which is the least hydrophobic, and characterized by higher compared to NP and OP steric hindrance and electrostatic repulsion.


Caregiver and provider perspectives on dual VA and Medicare Part D medication use in veterans with suspected dementia or cognitive impairment.
Many older veterans with dementia fill prescriptions through both Veterans Affairs (VA) and Medicare Part D benefits. Dual VA/Part D medication use may have unintended negative consequences in terms of prescribing safety and quality. We aimed to characterize benefits and drawbacks of dual VA/Part D medication use in veterans with dementia or cognitive impairment from the perspectives of caregivers and providers. This was a qualitative study based on semistructured telephone interviews of 2 groups: (1) informal caregivers accompanying veterans with suspected dementia or cognitive impairment to visits at a VA Geriatric Evaluation and Management clinic (n = 11) and (2) VA healthcare providers of veterans with dementia who obtained medications via VA and Part D (n = 12). We conducted semistructured telephone interviews with caregivers and providers about benefits and drawbacks of dual VA/Part D medication use. Interview transcripts were subjected to qualitative content analysis to identify key themes. Caregivers and providers both described cost and convenience benefits to dual VA/Part D medication use. Caregivers reported drawbacks including poor communication between VA and non-VA providers and difficulty managing medications from multiple systems. Providers reported potential safety risks including communication barriers, conflicting care decisions, and drug interactions. Results of this study allow for understanding of potential policy interventions to better manage dual VA/Part D medication use for older veterans with dementia or cognitive impairment at a time when VA is expanding access to non-VA care.


Structure of the heavy chain from strain 13 guinea pig immunoglobulin G1: isolation of cyanogen bromide fragments.
Guinea pig serum contains two isotypes of immunoglobulin G: IgG1 and IgG2. These immunoglobulins are antigentically distinct from each other and they mediate different biologic processes in the same guinea pig, although they share the ability to bind the same antigen. An attempt was made to study the primary structure of the gamma1 heavy chain from IgG1 in comparison with the largely known primary structure of the gamma-heavy chain from IgG2, with the aim of demarcating the structural differences between these molecules. IgG1 was isolated from the serum of immune strain 13 guinea pigs. Both IgG1 and the gamma1 chain were digested with CNBr. Nine fragments were isolated from both digests by gel filtration procedures before and after reductive cleavage of disulfide bonds. These fragments appear to account for the entire approximately 444 residues in the gamma1 chain. Amino acid composition data of CNBr fragments suggest that at least the amino terminal approximately 182 residues of the gamma1 and gamma2 chains are very similar. Two of the fragments which have been isolated have amino acid compositions suggesting their derivation from the "hinge" region and carboxyl terminus of the gamma1 chain.


Differential 3D Facial Recognition: Adding 3D to Your State-of-the-Art 2D Method.
Active illumination is a prominent complement to enhance 2D face recognition and make it more robust, e.g., to spoofing attacks and low-light conditions. In the present work we show that it is possible to adopt active illumination to enhance state-of-the-art 2D face recognition approaches with 3D features, while bypassing the complicated task of 3D reconstruction. The key idea is to project over the test face a high spatial frequency pattern, which allows us to simultaneously recover real 3D information plus a standard 2D facial image. Therefore, state-of-the-art 2D face recognition solution can be transparently applied, while from the high frequency component of the input image, complementary 3D facial features are extracted. Experimental results on ND-2006 dataset show that the proposed ideas can significantly boost face recognition performance and dramatically improve the robustness to spoofing attacks.


Sound scattering by several zooplankton groups. I. Experimental determination of dominant scattering mechanisms.
The acoustic scattering properties of live individual zooplankton from several gross anatomical groups have been investigated. The groups involve (1) euphausiids (Meganyctiphanes norvegica) whose bodies behave acoustically as a fluid material, (2) gastropods (Limacina retroversa) whose bodies include a hard elastic shell, and (3) siphonophores (Agalma okeni or elegans and Nanomia cara) whose bodies contain a gas inclusion (pneumatophore). The animals were collected from ocean waters off New England (Slope Water, Georges Bank, and the Gulf of Maine). The scattering properties were measured over parts or all of the frequency range 50 kHz to 1 MHz in a laboratory-style pulse-echo setup in a large tank at sea using live fresh specimens. Individual echoes as well as averages and ping-to-ping fluctuations of repeated echoes were studied. The material type of each group is shown to strongly affect both the overall echo level and pattern of the target strength versus frequency plots. In this first article of a two-part series, the dominant scattering mechanisms of the three animal types are determined principally by examining the structure of both the frequency spectra of individual broadband echoes and the compressed pulse (time series) output. Other information is also used involving the effect on overall levels due to (1) animal orientation and (2) tissue in animals having a gas inclusion (siphonophores). The results of this first paper show that (1) the euphausiids behave as weakly scattering fluid bodies and there are major contributions from at least two parts of the body to the echo (the number of contributions depends upon angle of orientation and shape), (2) the gastropods produce echoes from the front interface and possibly from a slow-traveling circumferential (Lamb) wave, and (3) the gas inclusion of the siphonophore dominates the echoes, but the tissue plays a role in the scattering and is especially important when analyzing echoes from individual animals on a ping-by-ping basis. The results of this paper serve as the basis for the development of acoustic scattering models in the companion paper [Stanton et al., J. Acoust. Soc. Am. 103, 236-253 (1998)].


Surgical and nonsurgical management of rotator cuff tears.
Although rotator cuff tears may be asymptomatic, symptomatic tears have had treatment varying from nonoperative therapy and debridement with subacromial decompression to operative repair. The benchmark for comparison should be nonoperative treatment when deciding to proceed to operative repair. This review will analyze the recent peer-reviewed publications describing results of operative repair, how this regimen differs with nonoperative treatment, and any outcome differences between operative and nonoperative treatment. Operative repair has consistently shown a higher rate of pain relief in patients (85%) and a better return of strength. Nonoperative treatment is expected to produce satisfactory relief of pain in only 50% of patients and no improvement in strength at long-term follow-up.


[Changes in the transcription capacity of the chromatin of nerve and liver cells exposed to morphine and opioids].
Morphine stimulates in vitro 3H-UTP incorporation into the nuclei of neurons of the superior cervical ganglion and into the hepatocyte nuclei of some rodents. This effect is dose-dependent and partially inhibited by naloxone. A synthetic analog of enkephalins, D-Ala2-Gly-ol5-enkephalin (DAGO), and beta-endorphin stimulate the incorporation of RNA precursors into pre-rRNA. Possible mechanisms of morphine action are discussed.


Common and unique cis-acting elements mediate xanthotoxin and flavone induction of the generalist P450 CYP321A1.
How polyphagous herbivores up-regulate their counterdefense genes in response to a broad range of structurally different allelochemicals remains largely unknown. To test whether this is accomplished by having more allelochemical-response elements or the similar number of functionally more diverse elements, we mapped out the cis-acting elements mediating the induction of the allelochemical-metabolizing CYP321A1 from the generalist Helicoverpa zea by xanthotoxin and flavone, two structurally distinct allelochemicals with very different encounter rate by this species. Seven xanthotoxin-responsive elements were localized by analyzing promoter activities of varying length of CYP321A1 promoter in H. zea fatbody cells. Compared with the 5 flavone-responsive elements mapped out previously, there are four common elements (1 essential element, 2 enhancers, and 1 negative element) mediating induction of CYP321A1 by both of the two allelochemicals. The remaining four elements (3 enhancers and 1 negative element), however, only regulate induction of CYP321A1 by either of the two allelochemicals. Co-administration of the two allelochemicals resulted in an induction fold that is significantly lower than the expected additive value of the two allelochemicals. These results indicate that xanthotoxin- and flavone-induced expressions of CYP321A1 are mediated mainly by the functionally more diverse common elements although the allelochemical-unique elements also play a role.


A simple reminder system improves venous thromboembolism prophylaxis rates and reduces thrombotic events for hospitalized patients1.
Compliance with venous thromboembolism (VTE) prophylaxis is poor. We sought to determine whether a simple electronic reminder applicable to all hospitalized patients would increase prophylaxis rates and reduce VTE rates. An electronic reminder was added to the electronic medical record admission note used by all services in our hospital. Prophylaxis, VTE and bleeding rates before and after implementation were compared. Data were analyzed with sas version 9.1. Among all adult medical and surgical patients admitted to our hospital during the time periods studied, 42.8% (1236/2888) before and 60.0% (1410/2350) after the reminder was added received appropriate prophylaxis as per American College of Chest Physicians (ACCP) guidelines (P < 0.001). The difference reached significance for both medical (51.0% vs. 68.9%; P < 0.001) and surgical (48.0% vs. 61.0%; P < 0.001) services. Fewer patients were diagnosed with VTE after our reminder was added (1.1% vs. 0.3%; P = 0.001), and there was a trend towards fewer bleeds (1.1% vs. 0.6%; P = 0.09). The presence of the reminder was an independent predictor for prophylaxis being given (odds ratio [OR] 1.92, 95% confidence interval [CI] 1.70-2.18; P < 0.001), and was independently associated with a decreased risk for VTE (OR 0.30, 95% CI 0.14-0.64; P = 0.003) after adjustment for other VTE risk factors. Adding an electronic reminder to the admission note improved prophylaxis rates and reduced VTE rates across services. The system is easily reproducible and applicable to other facilities. The improvement obtained was modest, so additional measures will probably be needed to optimize prophylaxis rates.


Deoxycholic acid mediates non-canonical EGFR-MAPK activation through the induction of calcium signaling in colon cancer cells.
Obesity and a western diet have been linked to high levels of bile acids and the development of colon cancer. Specifically, increased levels of the bile acid deoxycholic acid (DCA), an established tumor promoter, has been shown to correlate with increased development of colorectal adenomas and progression to carcinoma. Herein we investigate the mechanism by which DCA leads to EGFR-MAPK activation, a candidate mechanism by which DCA may promote colorectal tumorigenesis. DCA treated colon cancer cells exhibited strong and prolonged activation of ERK1/2 when compared to EGF treatment alone. We also showed that DCA treatment prevents EGFR degradation as opposed to the canonical EGFR recycling observed with EGF treatment. Moreover, the combination of DCA and EGF treatment displayed synergistic activity, suggesting DCA activates MAPK signaling in a non-canonical manner. Further evaluation showed that DCA treatment increased intracellular calcium levels and CAMKII phosphorylation, and that blocking calcium with BAPTA-AM abrogated MAPK activation induced by DCA, but not by EGF. Finally we showed that DCA-induced CAMKII leads to MAPK activation through the recruitment of c-Src. Taken together, we demonstrated that DCA regulates MAPK activation through calcium signaling, an alternative mechanism not previously recognized in human colon cancer cells. Importantly, this mechanism allows for EGFR to escape degradation and thus achieve a constitutively active state, which may explain its tumor promoting effects.


Effects of Cisplatin and its analogues on the permeability of human erythrocyte membrane.
The biomembrane is postulated as the initial target when Platinum(II) complexes attack cells. In this work, a spin-labeling ESR technique has been used to study the effects of cis-DCDP, cis-DBDP, cis-DIDP, trans-DCDP, and cis-DADP on the permeability of human erythrocyte membrane. We monitored the reduction processes of the ESR signal of a nitroxide spin label, (TEMPO), which leaks out through the membrane and is reduced by the external ascorbate. Our results indicate that cisplatin and its analogues can enhance the permeability of membranes to small moieties such as TEMPO and ascorbate, and the differences between these compounds are related to features of the leaving group. In addition, changes in the order parameter of 5DS spin label in membrane indicate that hydrolysis of these Pt(II) complexes result in membrane damage.


Structure-based discovery of nonopioid analgesics acting through the α2A-adrenergic receptor.
Because nonopioid analgesics are much sought after, we computationally docked more than 301 million virtual molecules against a validated pain target, the α2A-adrenergic receptor (α2AAR), seeking new α2AAR agonists chemotypes that lack the sedation conferred by known α2AAR drugs, such as dexmedetomidine. We identified 17 ligands with potencies as low as 12 nanomolar, many with partial agonism and preferential Gi and Go signaling. Experimental structures of α2AAR complexed with two of these agonists confirmed the docking predictions and templated further optimization. Several compounds, including the initial docking hit '9087 [mean effective concentration (EC50) of 52 nanomolar] and two analogs, '7075 and PS75 (EC50 4.1 and 4.8 nanomolar), exerted on-target analgesic activity in multiple in vivo pain models without sedation. These newly discovered agonists are interesting as therapeutic leads that lack the liabilities of opioids and the sedation of dexmedetomidine.


Revisiting the Engineering Roadmap of Nitrate/Nitrite-Dependent Anaerobic Methane Oxidation.
Nitrate/nitrite-dependent anaerobic oxidation of methane (n-DAMO) is a recently discovered process, which provides a sustainable perspective for simultaneous nitrogen removal and greenhouse gas emission (GHG) mitigation by using methane as an electron donor for denitrification. However, the engineering roadmap of the n-DAMO process is still unclear. This work constitutes a state-of-the-art review on the classical and most recently discovered metabolic mechanisms of the n-DAMO process. The versatile combinations of the n-DAMO process with nitrification, nitritation, and partial nitritation for nitrogen removal are also clearly presented and discussed. Additionally, the recent advances in bioreactor development are systematically reviewed and evaluated comprehensively in terms of methane supply, biomass retention, membrane requirement, startup time, reactor performance, and limitations. The key issues including enrichment and operation strategy for the scaling up of n-DAMO-based processes are also critically addressed. Moreover, the challenges inherent to implementing the n-DAMO process in practical applications, including application scenario recognition, GHG emission mitigation, and operation under realistic conditions, are highlighted. Finally, prospects as well as opportunities for future research are proposed. Overall, this review provides a roadmap for potential applications and further development of the n-DAMO process in the field of wastewater treatment.


New perspectives on nodule nitrogen assimilation in actinorhizal symbioses.
Nitrogen-fixing root nodules are plant organs specialised for symbiotic transfer of nitrogen and carbon between microsymbiont and host. The organisation of nitrogen assimilation, storage and transport processes is partitioned at the subcellular and tissue levels, in distinctive patterns depending on the symbiotic partners. In this review, recent advances in understanding of actinorhizal nodule nitrogen assimilation are presented. New findings indicate that Frankia within nodules of Datisca glomerata (Presl.) Baill. carries out both primary nitrogen assimilation and biosynthesis of arginine, rather than exporting ammonium. Arginine is a typical storage form of nitrogen in plant tissues, but is a novel nitrogen carrier molecule in root nodule symbioses. Thus Frankia within D. glomerata nodules exhibits considerable metabolic independence. Furthermore, nitrogen reassimilation is likely to take place in the host in the uninfected nodule cortical cells of this root nodule symbiosis, before amino acid export to host sink tissues via the xylem. The role of an augmented pericycle in carbon and nitrogen exchange in root nodules deserves further attention in actinorhizal symbiosis, and further highlights the importance of a comprehensive, structure-function approach to understanding function in root nodules. Moreover, the multiple patterns of compartmentalisation in relation to nitrogen flux within root nodules demonstrate the diversity of possible functional interactions between host and microsymbiont that have evolved in the nitrogen-fixing clade.


Dyslipidemia evaluated by brachial artery vasoactivity: a case study of eight patients.
Current National Cholesterol Education Program (NCEP) guidelines consider a desirable low-density lipoprotein cholesterol level to be < 100 mg/dl for an individual with coronary artery disease (CAD). To assess the affect of these cholesterol levels on endothelial cell functioning, flow-mediated brachial artery vasoactivity was measured non-invasively (7.5 MHz ultrasound) before and during Simvastatin (Zocor) therapy. Vasoactivity was expressed as a percent diameter change from baseline to hyperemic conditions and increased from 2.4 +/- 3.0% at baseline to 13.1 +/- 3.4% (p < .05) and 15.1 +/- 2.7% (p < .01) as cholesterol decreased from 243 +/- 24.7 to 221 +/- 27.3 and 213 +/- 22 mg/dl after three and six months, respectively, during therapy. LDL cholesterol subsequently decreased from 155 +/- 22.1 at baseline to 143 +/- 20.2 and 130 +/- 10.6 after three and six months of treatment, respectively. In conclusion, vasoactivity was found to correlate inversely with cholesterol levels (r = 0.44). This suggests that vasoactivity responds to changes in cholesterol levels and that endothelial function improves by lowering cholesterol levels.


Intestinal microsporidiosis in HIV infected patients with chronic diarrhea in Thailand.
Microsporidia have been recognized as emerging opportunistic agents affecting multiple organs. Intestinal microsporidiosis caused by Enterocytozoon bieneusi and Encephalitozoon intestinalis is a common disease which is associated with gastrointestinal symptoms, particularly in AIDS patients. So far, information on the frequency of this enteric disease in Thailand is not available. Therefore, the present study was undertaken to investigate the prevalence of intestinal microsporidiosis in HIV infected persons with chronic diarrhea. From 1995 to 1996, multiple diarrheal stool specimens were received and examined for the presence of the organism using Weber's modified trichrome staining method and transmission electron microscopy for confirmation. Twenty-two of 66 patients (33.3%) were positive for microsporidia which appeared as pink-red spores of 0.8-1.2 x 0.7-0.9 microm with the characteristic transverse or oblique band representing the coiled polar filament. Clinical features of these patients included chronic diarrhea (100%), weight loss (100%), abdominal pain (77%), fever (36%), vomiting (36%) and anorexia (18%). Transmission electron microscopic examination of fecal specimens from the 22 patients with positive staining results revealed E. bieneusi in 18 cases.


Stress fractures of the second metatarsal involving Lisfranc's joint in ballet dancers. A new overuse injury of the foot.
We reviewed the cases of four female ballet dancers with a stress fracture of a type that has not been reported previously. This fracture occurs in the proximal portion of the second metatarsal and involves the volar and medial aspects of Lisfranc's joint. A differential diagnosis of pain in the middle part of the foot in a dancer should include a consideration of this entity, which can be very difficult to diagnose on initial assessment. Oblique radiographs, tomograms, and a bone scan may be necessary to confirm the diagnosis. With early recognition and diagnosis, in three of the four patients the fracture healed with immobilization and modified training. One patient required surgical resection because of persistent non-union of the necrotic fracture fragment.


Therapeutic targets in the ARF tumor suppressor pathway.
One of the outstanding fundamental questions in cancer cell biology concerns how cells coordinate cellular growth (or macromolecular synthesis) with cell cycle progression and mitosis. Intuitively, rapidly dividing cells must have some control over these processes; otherwise cells would continue to shrink in volume with every passing cycle, similar to the cytoreductive divisions seen in the very early stages of embryogenesis. The problem is easily solved in unicellular organisms, such as yeast, as their growth rates are entirely dependent on nutrient availability. Multicellular organisms such as mammals, however, must have acquired additional levels of control, as nutrient availability is seldom an issue and the organism has a prodigious capacity to store necessary metabolites in the form of glycogen, lipids, and protein. Furthermore, the specific needs and specialized architecture of tissues must constrain growth for growth's sake; if not, the necessary function of the organ could be lost. While certainly a myriad of mechanisms for preventing this exist via initiating cell death (e.g. apoptosis, autophagy, necrosis), these all depend on some external cue, such as death signals, hypoxia, lack of nutrients or survival signals. However there must also be some cell autonomous method for surveying against inappropriate growth signals (such as oncogenic stress) that occur in a stochastic fashion, possibly as a result of random mutations. The ARF tumor suppressor seems to fulfill that role, as its expression is near undetectable in normal tissues, yet is potently induced by oncogenic stress (such as overexpression of oncogenic Ras or myc). As a result of induced expression of ARF, the tumor suppressor protein p53 is stabilized and promotes cell cycle arrest. Mutations or epigenetic alterations of the INK4a/Arf locus are second only to p53 mutations in cancer cells, and in some cancers, alterations in both Arf and p53 observed, suggesting that these two tumor suppressors act coordinately to prevent unwarranted cell growth and proliferation. The aim of this review is to characterize the current knowledge in the field about both p53-dependent and independent functions of ARF as well as to summarize the present models for how ARF might control rates of cell proliferation and/or macromolecular synthesis. We will discuss potential therapeutic targets in the ARF pathway, and some preliminary attempts at enhancing or restoring the activity of this important tumor suppressor.


Metabolic synergy: increasing biosynthetic capabilities by network cooperation.
Cooperation between organisms of different species is a widely observed phenomenon in biology, ranging from large scale systems such as whole ecosystems to more direct interactions like symbiotic relationships. In the present work, we explore inter-species cooperations on the level of metabolic networks. For our analysis, we extract 447 organism specific metabolic networks from the KEGG database [7] and assess their biosynthetic capabilities by applying the method of network expansion [5]. We simulate the cooperation of two organisms by unifying their metabolic networks and introduce a measure, the gain Gamma, quantifying the amount by which the biosynthetic capability of an organism is enhanced due to the cooperation with another species. For all theoretically possible pairs of organisms, this synergetic effect is determined and we systematically analyze its dependency on the dissimilarities of the interacting partners. We describe these dissimilarities by two different distance measures, where one is based on structural, the other on evolutionary differences. With the presented method, we provide a conceptional framework to study the metabolic effects resulting from an interaction of different species. We outline possible enhancements of our analysis: by defining more realistic interacting networks and applying alternative structural investigation methods, our concept can be used to study specific symbiotic and parasitic relationships and may help to understand the global interplay of metabolic pathways over the boundary of organism specific systems.


Laboratory and clinical assessment of pneumatically driven intracorporeal lithotripsy.
A pneumatically driven intracorporeal lithotripter (the Swiss Lithoclast) has recently been approved for use in the United States. We compared its performance in vitro with ultrasonic, electrohydraulic and laser lithotripsy devices using a standard plaster-of-Paris stone phantom. The probe sizes and output settings were identical to those used during clinical treatment. The fragmentation efficiency index (measured as the lithotripsy time needed to reduce the stone phantom to particles <2 mm divided by the initial stone weight) ranged from 5.0 to 8.5 min/g of stone mass, with this value increasing from pneumatic to electrohydraulic to laser and to ultrasonic lithotripsy. We also performed an objective study in a swine model, which showed no adverse consequence of pneumatic lithotripsy. Finally, we evaluated our initial 41 patients who had undergone pneumatic stone fragmentation. We treated 8 patients having 11 renal calculi, 30 patients having 37 ureteral calculi, and 3 patients having 6 bladder calculi employing pneumatic probes ranging in size from 0.8 to 2.0 mm. Stone fragmentation was successful in a single session in 95% of the ureteral calculi and 100% of both renal and bladder calculi. Stone analysis in 23 patients revealed 17 (74%) calcium oxalate monohydrate and 1 (4%) cystine calculi. Our clinical and laboratory assessment of this newly developed pneumatic lithotripsy device further validates its efficacy in fragmenting stone of all compositions and its overall safety associated with clinical application.


The EBI Search engine: providing search and retrieval functionality for biological data from EMBL-EBI.
The European Bioinformatics Institute (EMBL-EBI-https://www.ebi.ac.uk) provides free and unrestricted access to data across all major areas of biology and biomedicine. Searching and extracting knowledge across these domains requires a fast and scalable solution that addresses the requirements of domain experts as well as casual users. We present the EBI Search engine, referred to here as 'EBI Search', an easy-to-use fast text search and indexing system with powerful data navigation and retrieval capabilities. API integration provides access to analytical tools, allowing users to further investigate the results of their search. The interconnectivity that exists between data resources at EMBL-EBI provides easy, quick and precise navigation and a better understanding of the relationship between different data types including sequences, genes, gene products, proteins, protein domains, protein families, enzymes and macromolecular structures, together with relevant life science literature.


Sodium alginate as a novel therapeutic option in experimental colitis.
The potential therapeutic effect of low-viscosity sodium alginate (LVA) was studied in a rat model of acute colitis induced by intracolonic administration of acetic acid. This experimental model produced a significant ulcerative colitis. Induction of colitis also significantly enhanced the serum and colonic mucosal cytokine (IL-6 and TNF-alpha) and eicosanoid (LTB4 and PGE2) levels, which paralleled with the severity of colitis. LVA solution was administered orally as drinking water at concentration of 0.5% (W/V) for 1 week. The tolerability and inhibitory effect of LVA on matrix metalloproteinase-2 (MMP-2) were tested using WEHI-164 cell line and zymography method. The results showed that LVA therapy is able to significantly reduce colonic damage score, histological lesion, serum and colonic mucosal IL-6, TNF-alpha, LTB4 and PGE2 levels in treated group compared with nontreated controls. Moreover, in vitro examinations revealed that treatment with LVA could diminish MMP-2 activity. It is concluded that LVA is able to suppress acetic acid-induced colitis in rats. Some of the action of LVA may be associated with its inhibitory effects on cytokine and eicosanoid production and MMP-2 activity. Our data suggest that LVA could potentially be a novel therapeutic option for inflammatory bowel disease.


Development of telehealth principles and guidelines for older adults: A modified Delphi approach.
The COVID-19 pandemic elevated telehealth as a prevalent care delivery modality for older adults. However, guidelines and best practices for the provision of healthcare via telehealth are lacking. Principles and guidelines are needed to ensure that telehealth is safe, effective, and equitable for older adults. The Collaborative for Telehealth and Aging (C4TA) composed of providers, experts in geriatrics, telehealth, and advocacy, developed principles and guidelines for delivering telehealth to older adults. Using a modified Delphi process, C4TA members identified three principles and 18 guidelines. First, care should be person-centered; telehealth programs should be designed to meet the needs and preferences of older adults by considering their goals, family and caregivers, linguistic characteristics, and readiness and ability to use technology. Second, care should be equitable and accessible; telehealth programs should address individual and systemic barriers to care for older adults by considering issues of equity and access. Third, care should be integrated and coordinated across systems and people; telehealth should limit fragmentation, improve data sharing, increase communication across stakeholders, and address both workforce and financial sustainability. C4TA members have diverse perspectives and expertise but a shared commitment to improving older adults' lives. C4TA's recommendations highlight older adults' needs and create a roadmap for providers and health systems to take actionable steps to reach them. The next steps include developing implementation strategies, documenting current telehealth practices with older adults, and creating a community to support the dissemination, implementation, and evaluation of the recommendations.


Implementation of a multidisciplinary guideline improves preterm infant admission temperatures.
Hypothermia is a common problem in preterm infants immediately following delivery.Local problem:The rate of admission hypothermia in our neonatal intensive care unit (NICU) was above the rate of comparable NICUs in the Vermont Oxford Network. To reduce the rate of preterm admission hypothermia, a quality improvement (QI) project was implemented, utilizing the plan-do-study-act (PDSA) methodology. A guideline for delivery room thermoregulation management in <35-week infants at the University of Virginia was created and put into practice by a multidisciplinary team. Clinical practice changes in the guideline included: increasing operating room temperatures, obtaining a 10-min axillary temperature, using an exothermic mattress for all infants <35 weeks, and using a polyethylene wrap for infants <32 weeks. The baseline rate of hypothermia (<36.5 °CC) was 63%. Three PDSA cycles data were completed on 168 consecutive preterm births. The post-implementation rate of hypothermia (<36.5 °C) was reduced to 30% (P<0.001). The incidence of moderate hypothermia (< 36 °C) was reduced from a baseline of 29% to a rate of 9% (P<0.001). Use of a multidisciplinary guideline to increase preterm NICU admission temperatures resulted in a decrease in hypothermic infants.


Clinical characteristics of Martorell hypertensive ischaemic leg ulcer.
We sought to characterise the clinical picture of Martorell hypertensive ischaemic leg ulcer (HYTILU) by describing the ulcer borders with three clinical features: 'the red lipstick sign'; purple border; and livedo racemosa. We also aimed to characterise comorbidities and determinants of healing time. A single-centre, retrospective cohort study was conducted between 2015-2020. We scrutinised ulcer photographs for relevant clinical signs. Data on comorbidities, medication and ulcer treatments, as well as method of diagnosis and healing time, were collected from patients' electronic medical records. In total, 38 female patients and 31 male patients (mean age 73 years) were assessed, with a mean follow-up time of 174 days. The 'red lipstick-like' margin covered 0-50% of the ulcer margin in 56.5% of the ulcers, and 51-100% of the ulcer margin in 43.5% of the ulcers. Purple border or livedo racemosa was observed in 70.5% of the ulcers. All patients had hypertension and 52.2% of patients had type 2 diabetes. A heavy cardiovascular disease burden and frequent concomitant vascular pathologies were found. Infections requiring systemic antibiotics, ulcer size and duration of symptoms before diagnosis were strongly associated with healing time. We also found that use of systemic corticosteroids and severity of hypertension (measured by the number of antihypertensive medications used) delayed healing. Our data suggest that 'the red lipstick sign' could be a novel diagnostic feature in HYTILUs alongside purple border, livedo racemosa and necrotic/fibrinous ulcer bed. The results also elucidated HYTILU comorbidities, and showed that infections and delay in diagnosis impeded healing.


Perspectives on induced abortion among Palestinian women: religion, culture and access in the occupied Palestinian territories.
Induced abortion is an important public health issue in the occupied Palestinian territories (OPT), where it is illegal in most cases. This study was designed to elicit the views of Palestinian women on induced abortion given the unique religious, ethical and social challenges in the OPT. Sixty Palestinian women were interviewed on their perceptions of the religious implications, social consequences and accessibility of induced abortions in the OPT at Al-Makassed Islamic Charitable Hospital in East Jerusalem. Themes arising from the interviews included: the centrality of religion in affecting women's choices and views on abortion; the importance of community norms in regulating perspectives on elective abortion; and the impact of the unique medico-legal situation of the OPT on access to abortion under occupation. Limitations to safe abortion access included: legal restrictions; significant social consequences from the discovery of an abortion by one's community or family; and different levels of access to abortion depending on whether a woman lived in East Jerusalem, the West Bank, or Gaza. This knowledge should be incorporated to work towards a legal and medical framework in Palestine that would allow for safe abortions for women in need.


A Scientific Symposium "Artificial Organs Today: From in Vitro Assessment to Human Therapies" on the Occasion of the Celebration of the 40th Anniversary of the Macedonian Academy of Sciences and Arts.
On the occasion of the celebration of the 40th anniversary of the Macedonian Academy of Sciences and Arts (MASA) the Macedonian Society of Nephrology, Dialysis, Transplantation and Artificial Organs (MSNDTAO) organized a one day symposium titled: "Artificial Organs Today: From in vitro assessment to human therapies", on September 28, 2007 at the Macedonian Academy of Sciences and Arts. The following sessions were held: Artificial Organs in Front of the Ageing Populations; System Requirements for Artificial Organ Technology; Tools for Artificial Organ Technology; Extracorporeal Blood Circuits in Organ Replacement Therapies; Treatment Options for Blood Purification Therapies. Presentation were delivered by: Horst Klinkmann, Germany; Jörg Vienken, Germany; Jens Hartmann, Austria; Udo Losert, Austria; Jan Wojcicki, Poland; Helmut Mann and Siegfried Stiller, Germany; Beat Walpoth, Switzerland; Juan F. del Cañizo, Spain; Borce Georgievski, Macedonia; Bernd Stegmayr, Sweden; Momir Polenakovic, Macedonia; Petar Kes, Croatia; Aleksandar Sikole, Macedonia; Wolfgang Ramlow, Germany; Dieter Falkenhagen, Austria. These meetings are significant for the application and development of the artificial organs in patients.


Understanding API-polymer proximities in amorphous stabilized composite drug products using fluorine-carbon 2D HETCOR solid-state NMR.
A simple and robust method for obtaining fluorine-carbon proximities was established using a (19)F-(13)C heteronuclear correlation (HETCOR) two-dimensional (2D) solid-state nuclear magnetic resonance (ssNMR) experiment under magic-angle spinning (MAS). The method was applied to study a crystalline active pharmaceutical ingredient (API), avagacestat, containing two types of fluorine atoms and its API-polymer composite drug product. These results provide insight into the molecular structure, aid with assigning the carbon resonances, and probe API-polymer proximities in amorphous spray dried dispersions (SDD). This method has an advantage over the commonly used (1)H-(13)C HETCOR because of the large chemical shift dispersion in the fluorine dimension. In the present study, fluorine-carbon distances up to 8 Å were probed, giving insight into the API structure, crystal packing, and assignments. Most importantly, the study demonstrates a method for probing an intimate molecular level contact between an amorphous API and a polymer in an SDD, giving insights into molecular association and understanding of the role of the polymer in API stability (such as recrystallization, degradation, etc.) in such novel composite drug products.


Fluorescent fibrous mats assembled with self-propagating probes for visual sensing of hydrogen peroxide and choline.
Challenges remain in the facile, rapid and sensitive detection of substances at ultralow levels. In the current study, visual sensors of hydrogen peroxide (H2O2) and choline are developed via the integration of an ultrafine fibrous substrate and self-propagating and aggregation-induced emission (AIE) probes. Self-immolative probes (SIPs) composed of phenylboronic acid triggers and choline units are grafted on electrospun polyethylene terephthalate (PET) fibers, followed by electrostatic adsorption of tetraphenylethene derivatives (TPE-SO3) to obtain fluorescent PET-Ch/TPE fibers. Choline oxidase (ChOX) is immobilized on polystyrene-co-maleic anhydride (PSMA) fibers to obtain PSMA-ChOX, followed by assembly into PET-Ch/TPE@PSMA-ChOX composite mats. The presence of H2O2 initiates the cleavage of phenylboronic acid triggers in SIPs to release choline and choline/TPE complexes from PET-Ch/TPE fibers. The released choline is oxidized by PSMA-ChOX fibers to generate H2O2 that then activates a cascade of self-propagating reactions until the release of all choline/TPE complexes, leading to the alleviation of AIE effect and gradual fluorescence fading of fibrous mats. Thus, the hydrogen peroxide and choline concentrations can be read out from the fluorescence fading time of fibrous mats with a detection limit of 0.5 μM H2O2 within 30 min, providing potential self-test devices for a real-time, naked-eye and sensitive detection of bioactive substances.


Short-day potentiation of low temperature-induced gene expression of a C-repeat-binding factor-controlled gene during cold acclimation in silver birch.
Development of winter hardiness in trees is a two-stage process involving sequential perception of distinct environmental cues, short-day (SD) photoperiod and low temperature (LT). We have shown that both SD and LT are recognized by leaves of silver birch (Betula pendula cv Roth) leading to increased freezing tolerance, and thus leaves can be used as an experimental model to study the physiological and molecular events taking place during cold acclimation. To obtain a molecular marker for the acclimation process in birch we cloned a gene, designated Bplti36, encoding a 36-kD acidic SK2 type of dehydrin. The gene was responsive to LT, drought, salt, and exogenous abscisic acid. This responsiveness to abiotic stresses and abscisic acid was retained when Bplti36 was introduced to Arabidopsis (Arabidopsis thaliana). The LT induction of the gene appeared to be under the control of the C-repeat-binding factor pathway as suggested by the presence of several C-repeat/dehydration-responsive element/LT-responsive elements in the Bplti36 promoter and its constitutive expression in C-repeat-binding factor overproducing Arabidopsis. In birch SD photoperiod at normal-growth temperature did not result in significant induction of Bplti36. However, preexposure to SD followed by LT treatment resulted in a remarkable increase in Bplti36 transcript accumulation as compared to LT-treated plants grown at long-day photoperiod. This suggests that SD photoperiod potentiates the LT response by conditioning the leaf tissue to be more responsive to the LT stimulus.


Structure and special chemical reactivity of interface-stabilized cerium oxide nanolayers on TiO2(110).
Novel interface-stabilized ceria nanophases have been grown on TiO2(110) by physical vapor deposition. At low coverage, dumbbell nanostructures constituted by reconstructed titania and ceria clusters are formed, while long range ordered nanoxides can be obtained by increasing the ceria dose. Scanning tunneling microscopy and photoemission spectroscopy were used to characterize the electronic properties of the films, showing that the TiO2 substrate can effectively stabilize ceria in reduced form over a wide range of experimental conditions. Epitaxial coupling is a very useful tool for tuning the chemical properties of mixed oxide systems. The special electronic properties of the films have a direct counterpart in the chemical activity, which has been investigated by temperature programmed desorption using methanol as a probe molecule. The experimental results indicate an exceptional activity of the ceria-titania interface in the selective dehydration of methanol to formaldehyde at an unprecedented low temperature (330 K).


Conferral of enhanced natural killer cell function by KIR3DS1 in early human immunodeficiency virus type 1 infection.
A flurry of recent reports on the role of activating and inhibitory forms of the killer cell immunoglobulin-like receptors (KIR) in natural killer (NK) cell activity against human immunodeficiency virus type 1 (HIV-1) have yielded widely divergent results. The role of the activating NK receptor encoded by the KIR3DS1 allele and its putative ligands, members of the HLA class I Bw4Ile80 cluster, in early HIV-1 disease is controversial. We selected 60 treatment-naïve adults for study from the OPTIONS cohort of individuals with early HIV-1 infection in San Francisco. We performed NK cell functional assays measuring gamma interferon (IFN-gamma) and CD107a expression by NK cells in the unstimulated state and after stimulation by the major histocompatibility complex class I-deficient 721.221 B-lymphoblastoid cell line. In addition, we measured CD38 expression (a T-cell activation marker) on T and NK cells. Persons who have at least one copy of the KIR3DS1 gene had higher IFN-gamma and CD107a expression in the unstimulated state compared to those who do not possess this gene. After stimulation, both groups experienced a large induction of IFN-gamma and CD107a, with KIR3DS1 carriers achieving a greater amount of IFN-gamma expression. Differences in effector activity correlating with KIR3DS1 were not attributable to joint carriage of HLA Bw4Ile80 and KIR3DS1. We detected a partial but not complete dependence of KIR3DS1 on the members of B*58 supertype (B*57 and B*58) leading to higher NK cell function. Possessing KIR3DS1 was associated with lower expression of CD38 on both CD8(+) T and NK cells and with a loss or weakening of the known strong associations between CD8(+) T-cell expression of CD38 mean fluorescence intensity and the HIV-1 viral load. We observed that possessing KIR3DS1 was associated with higher NK cell effector functions in early HIV-1 disease, despite the absence of HLA Bw4Ile80, a putative ligand of KIR3DS1. Carriage of KIR3DS1 was associated with diminished CD8(+) T-cell activation, as determined by expression of CD38, and a disruption of the traditional relationship between viral load and activation in HIV-1 disease, which may lead to better clinical outcomes for these individuals.


Flavokawain B, a kava chalcone, induces apoptosis via up-regulation of death-receptor 5 and Bim expression in androgen receptor negative, hormonal refractory prostate cancer cell lines and reduces tumor growth.
Limited success has been achieved in extending the survival of patients with metastatic and hormone-refractory prostate cancer (HRPC). There is a strong need for novel agents in the treatment and prevention of HRPC. We have shown that flavokawain B (FKB), a kava chalcone, is about 4- to 12-fold more effective in reducing the cell viabilities of androgen receptor (AR)-negative, HRPC cell lines DU145 and PC-3 than AR-positive, hormone-sensitive prostate cancer cell lines LAPC4 and LNCaP, with minimal effect on normal prostatic epithelial and stromal cells. FKB induces apoptosis with an associated increased expression of proapoptotic proteins: death receptor-5, Bim and Puma and a decreased expression of inhibitors of apoptosis protein: XIAP and survivin. Among them, Bim expression was significantly induced by FKB as early as 4 hr of the treatment. Knockdown of Bim expression by short-hairpin RNAs attenuates the inhibitory effect on anchorage-dependent and -independent growth and caspase cleavages induced by FKB. These findings suggest that the effect of FKB, at least in part, requires Bim expression. In addition, FKB synergizes with TRAIL for markedly enhanced induction of apoptosis. Furthermore, FKB treatment of mice bearing DU145 xenograft tumors results in tumor growth inhibition and increases Bim expression in tumor tissues. Together, these results suggest robust mechanisms for FKB induction of apoptosis preferentially for HRPC and the potential usefulness of FKB for prevention and treatment of HRPC in an adjuvant setting.


Predictive regularity representations in violation detection and auditory stream segregation: from conceptual to computational models.
Predictive accounts of perception have received increasing attention in the past 20 years. Detecting violations of auditory regularities, as reflected by the Mismatch Negativity (MMN) auditory event-related potential, is amongst the phenomena seamlessly fitting this approach. Largely based on the MMN literature, we propose a psychological conceptual framework called the Auditory Event Representation System (AERS), which is based on the assumption that auditory regularity violation detection and the formation of auditory perceptual objects are based on the same predictive regularity representations. Based on this notion, a computational model of auditory stream segregation, called CHAINS, has been developed. In CHAINS, the auditory sensory event representation of each incoming sound is considered for being the continuation of likely combinations of the preceding sounds in the sequence, thus providing alternative interpretations of the auditory input. Detecting repeating patterns allows predicting upcoming sound events, thus providing a test and potential support for the corresponding interpretation. Alternative interpretations continuously compete for perceptual dominance. In this paper, we briefly describe AERS and deduce some general constraints from this conceptual model. We then go on to illustrate how these constraints are computationally specified in CHAINS.


Effectiveness of a guided online primary care intervention for fear of cancer recurrence: A randomised controlled trial.
Fear of Cancer Recurrence (FCR) is highly prevalent among cancer survivors and leads to decreased quality of life and increased healthcare costs. We assessed the effectiveness of a guided online primary care intervention for FCR, compared to waiting list. In this RCT, participants were recruited online and randomised 1:1. All adults who finished successful curative cancer treatment between 3 months and 10 years ago, wanted support for FCR, and had sufficient Dutch skills were eligible. The intervention consisted of a 10-week online programme and three to five video calling sessions with a trained mental health worker. After 6 months, the control group received the same intervention. The primary outcome was the difference between the groups in the change in FCR severity from baseline (T0) to 6 months (T2), measured online with the short form of the Fear of Cancer Recurrence Inventory. One hundred and seventy-three participants were enroled and randomised to the intervention (n = 86) or control group (n = 87). FCR severity dropped 2.1 points more in the intervention group than in the control group (2.7 points (SD = 3.9) versus 0.6 points (SD = 3.6), t(154) = 3.4, p = 0.0007). General mental well-being also improved significantly in the intervention group and remained stable in the control group. These improvements remained at 10 months follow up. This easily accessible and relatively inexpensive intervention effectively reduces FCR and has potential to replace or precede existing more intensive psychological treatments, improving patients' access to care. The trial was prospectively registered in the Netherlands Trial Register on 25-02-2019 with number NL7573.


Caring for children with sleep problems.
Sleep disturbances are common in infants and children. Sleep disturbances in children not only disrupt the child and family but also impact parental and child well-being, daytime functioning, and behavior. Pediatric nurses care for the individual child as well as their family members. Understanding the importance of healthy sleep habits and the implications of inadequate sleep on child behavior and family-peer interactions provides nurses an opportunity to decrease family stress and increase positive coping, adaptation, and family function. Common types of sleep problems are presented, and recommendations for screening tools are included to help nurses better assess sleep problems in children and make appropriate referrals.


Current issues in the diagnosis and management of blood culture-negative infective and non-infective endocarditis.
Diagnosis and management of blood culture-negative endocarditis constitute a formidable clinical challenge and a systemic approach is necessary for a successful outcome. Blood cultures are negative in endocarditis due mainly to preceding antibiotic administration or to fastidious slow-growing organisms. Less so, non-infective endocarditis is a paraneoplastic manifestation or may occur in association with autoimmune diseases. When the clinical diagnosis is contemplated and cultures and serologies are negative, histologic and molecular examination of the removed valve tissue may confirm the diagnosis. Treatment with antibiotics is often warranted and valve replacement remains appropriate for patients with heart failure or irreversible structural damage.


Effect of strong directional selection on weakly selected mutations at linked sites: implication for synonymous codon usage.
The fixation of weakly selected mutations can be greatly influenced by strong directional selection at linked loci. Here, I investigate a two-locus model in which weakly selected, reversible mutations occur at one locus and recurrent strong directional selection occurs at the other locus. This model is analogous to selection on codon usage at synonymous sites linked to nonsynonymous sites under strong directional selection. Two approximations obtained here describe the expected frequency of the weakly selected preferred alleles at equilibrium. These approximations, as well as simulation results, show that the level of codon bias declines with an increasing rate of substitution at the strongly selected locus, as expected from the well-understood theory that selection at one locus reduces the efficacy of selection at linked loci. These solutions are used to examine whether the negative correlation between codon bias and nonsynonymous substitution rates recently observed in Drosophila can be explained by this hitchhiking effect. It is shown that this observation can be reasonably well accounted for if a large fraction of the nonsynonymous substitutions on genes in the data set are driven by strong directional selection.


The Type VI Secretion System Engages a Redox-Regulated Dual-Functional Heme Transporter for Zinc Acquisition.
The type VI secretion system was recently reported to be involved in zinc acquisition, but the underlying mechanism remains unclear. Here, we report that Burkholderia thailandensis T6SS4 is involved in zinc acquisition via secretion of a zinc-scavenging protein, TseZ, that interacts with the outer membrane heme transporter HmuR. We find that HmuR is a redox-regulated dual-functional transporter that transports heme iron under normal conditions but zinc upon sensing extracellular oxidative stress, triggered by formation of an intramolecular disulfide bond. Acting as the first line of defense against oxidative stress, HmuR not only guarantees an immediate response to the changing environment but also provides a fine-tuned mechanism that allows a gradual response to perceived stress. The T6SS/HmuR-mediated active zinc transport system is also involved in bacterial virulence and contact-independent bacterial competition. We describe a sophisticated bacterial zinc acquisition mechanism affording insights into the role of metal ion transport systems.


The effects of rest and treatment following sport-related concussion: a systematic review of the literature.
To evaluate the evidence for rest, treatment, and rehabilitation following sport-related concussion (SRC). PubMed, CINAHL, PsychInfo, Cochrane Controlled Trials Registers, Health STAR, Sport Discus, EMBASE, Web of Science, and ProQuest. Articles were included if they met the following criteria: original research, reported SRC as a source of injury, and evaluated the effect of rest or treatment. Study design, participants, treatment, outcome measures, and key findings. Three studies met the inclusion criteria for evaluating the effects of rest and twelve for treatment. Low-intensity aerobic exercise may be of benefit. The current evidence evaluating the effect of rest and treatment following SRC is sparse. An initial period of rest may be of benefit. Low-level exercise and multimodal physiotherapy may be of benefit for those who are slow to recover. There is a strong need for high level studies evaluating the effects of rest and treatment following SRC.


Identification of ecdysone 22-long-chain fatty acyl esters in newly laid eggs of the cattle tick Boophilus microplus.
The five major apolar ecdysone esters present in newly laid eggs of the cattle tick Boophilus microplus have been purified by h.p.l.c. The quantities of the apolar esters present in the eggs were increased by administration of ecdysone to the mature females. G.c.-m.s. analysis, as their methyl esters, of the fatty acids released from the apolar ecdysone derivatives by alkali, coupled with positive-ion fast-atom-bombardment m.s. of the intact ecdysone esters, showed that the compounds consisted of a series of fatty acyl esters of ecdysone. The position of esterification of the ecdysone was established by p.m.r. spectroscopy. The combined data show that the novel apolar derivatives of ecdysone consist of the 22-palmitate, -palmitoleate, -stearate, -oleate, and -linoleate esters respectively. Confirmation was obtained by comparison with synthetic ecdysone 22-palmitate. The significance of the ecdysone fatty acyl esters as a possible source of free hormone during embryogenesis is discussed.