Substrate binding mechanism of a type I extradiol dioxygenase.
A meta-cleavage pathway for the aerobic degradation of aromatic hydrocarbons is catalyzed by extradiol dioxygenases via a two-step mechanism: catechol substrate binding and dioxygen incorporation. The binding of substrate triggers the release of water, thereby opening a coordination site for molecular oxygen. The crystal structures of AkbC, a type I extradiol dioxygenase, and the enzyme substrate (3-methylcatechol) complex revealed the substrate binding process of extradiol dioxygenase. AkbC is composed of an N-domain and an active C-domain, which contains iron coordinated by a 2-His-1-carboxylate facial triad motif. The C-domain includes a β-hairpin structure and a C-terminal tail. In substrate-bound AkbC, 3-methylcatechol interacts with the iron via a single hydroxyl group, which represents an intermediate stage in the substrate binding process. Structure-based mutagenesis revealed that the C-terminal tail and β-hairpin form part of the substrate binding pocket that is responsible for substrate specificity by blocking substrate entry. Once a substrate enters the active site, these structural elements also play a role in the correct positioning of the substrate. Based on the results presented here, a putative substrate binding mechanism is proposed.


Oncolytic parvoviruses: from basic virology to clinical applications.
Accumulated evidence gathered over recent decades demonstrated that some members of the Parvoviridae family, in particular the rodent protoparvoviruses H-1PV, the minute virus of mice and LuIII have natural anticancer activity while being nonpathogenic to humans. These studies have laid the foundations for the launch of a first phase I/IIa clinical trial, in which the rat H-1 parvovirus is presently undergoing evaluation for its safety and first signs of efficacy in patients with glioblastoma multiforme. After a brief overview of the biology of parvoviruses, this review focuses on the studies which unraveled the antineoplastic properties of these agents and supported their clinical use as anticancer therapeutics. Furthermore, the development of novel parvovirus-based anticancer strategies with enhanced specificity and efficacy is discussed, in particular the development of second and third generation vectors and the combination of parvoviruses with other anticancer agents. Lastly, we address the key challenges that remain towards a more rational and efficient use of oncolytic parvoviruses in clinical settings, and discuss how a better understanding of the virus life-cycle and of the cellular factors involved in virus infection, replication and cytotoxicity may promote the further development of parvovirus-based anticancer therapies, open new prospects for treatment and hopefully improve clinical outcome.


Synergistic antibacterial brilliant blue/reduced graphene oxide/quaternary phosphonium salt composite with excellent water solubility and specific targeting capability.
A water-soluble brilliant blue/reduced graphene oxide/tetradecyltriphenylphosphonium bromide composite (BB-rGO-TTP) was prepared by using noncovalent brilliant blue-functionalized reduced graphene oxide (BB-rGO) as the tetradecyltriphenylphosphonium bromide (TTP) carrier. Antibacterial performance of this novel composite was investigated for both Gram-positive and Gram-negative bacteria. The results showed that the novel BB-rGO-TTP, combing the advantages of graphene and TTP, displayed excellent synergistic antibacterial activity, specific targeting capability, water solubility, and mild cytotoxicity, suggesting the great potential application as sprayable graphene-based antibacterial solutions.


Loading Rate Has Little Influence on Tendon Fascicle Mechanics.
Mechanically, tendons behave like springs and store energy by stretching in proportion to applied stress. This relationship is potentially modified by the rate at which stress is applied, a phenomenon known as viscosity. Viscoelasticity, the combined effects of elasticity and viscosity, can affect maximum strain, the amount of stored energy, and the proportion of energy recovered (resilience). Previous studies of tendons have investigated the functional effects of viscoelasticity, but not at the intermediate durations of loading that are known to occur in fast locomotor events. In this study, we isolated tendon fascicles from rat tails and performed force-controlled tensile tests at rates between ∼10 MPa s-1 to ∼80 MPa s-1. At high rates of applied stress, we found that tendon fascicles strained less, stored less energy, and were more resilient than at low rates of stress (p = 0.007, p = 0.040, and p = 0.004, respectively). The measured changes, however, were very small across the range of strain rates studied. For example, the average strain for the slowest loading rate was 0.637% while it was 0.614% for the fastest loading. We conclude that although there is a measurable effect of loading rate on tendon mechanics, the effect is small and can be largely ignored in the context of muscle-actuated locomotion, with the possible exception of extreme muscle-tendon morphologies.


Rescue of mitomycin C- or psoralen-inactivated Micrococcus radiodurans by additional exposure to radiation or alkylating agents.
The processing of damaged DNA was altered in a mitomycin C-sensitive mutant (mtcA) of Micrococcus radiodurans. Even though the mutant retained resistance to 254-nm UV radiation, it did not, in contrast to the wild-type strain, show any excessive DNA degradation or cell death when incubated with chloramphenicol after sublethal doses of either UV light or mitomycin C. The results suggest the constitutive synthesis of an enzyme system responsible for wild-type proficiency in the repair of mitomycin C-induced damage. An alternative system able to repair damage caused by mitomycin C was demonstrated in the mtcA background. In this strain, additional damage inflicted upon the cellular DNA effected a massive rescue of cells previously inactivated by mitomycin C. Rescue was provoked by ionizing radiation, by UV light, or by simple alkylating agents. Cells treated with psoralen plus near-UV radiation could be rescued only when inactivation was due primarily to psoralen-DNA interstrand cross-links rather than to monoadducts. The rescue of inactivated cells was prevented in the presence of chloramphenicol. These results can be interpreted most readily in terms of an alternative repair system able to overcome DNA interstrand cross-links produced by mitomycin C or psoralen plus near-UV light, but induced only by the more abundant number of damages produced by radiation or simple alkylating agents.


Downsizing proteolytic digestion and analysis using dispenser-aided sample handling and nanovial matrix-assisted laser/desorption ionization-target arrays.
An efficient technique for enzymatic digestion of proteins in nanovial arrays and identification by peptide mass fingerprinting using matrix-assisted laser desorption/ionization (MALDI-MS) is presented in this work. Through dispensing of a protein solution with simultaneous evaporation the protein (substrate) is concentrated up to 300 times in-vial. At higher substrate concentrations the catalytic turnover numbers increase according to the Michaelis-Menten kinetics. Therefore, the dispenser-aided nanodigestion is valuable for identification of low-level proteins (10 nM-500 nM) as well as for automatic high efficiency digestions performed in 0.2-10 min. As an example of low-level protein identification, a 10 nM solution of lysozyme C was unambiguously identified after 5 min of nanodigestion. Moreover, only 30 s nanodigestion was sufficient to identify hemoglobin (10 microM), exemplifying the fast catalysis of the nanodigestion technique. The developed silicon flow-through piezoelectric dispenser is adapted for low-volume and preconcentrated samples in the nL-microL range and provides fast, accurate and contact-free sample positioning into the nanovials. In this work, the properties of the nanodigestion concept regarding proteins of different characteristics are explored. Furthermore, the potential of automated protein identification using precoated proteolytic nanovial-arrays is demonstrated.


Sonication-laser-excited time-resolved Shpol'skii spectrometry: a facile method for the direct screening of fifteen priority pollutants in soil samples.
A unique method for screening polycyclic aromatic hydrocarbons (PAH) in soil samples is reported. Milligrams of soil are sonicated into the vial of a cryogenic fiber-optic probe with 250 microL of n-octane. Pollutants partition into the layer of extracting solvent and the cryogenic probe is then used for fluorescence measurements at liquid helium temperature (4.2 K). N-octane provides PAH with the appropriate Shpol'skii matrix to record highly resolved spectra and characteristic fluorescence lifetimes. Unambiguous PAH determination is directly performed from the layer of extracting solvent with no need for previous PAH separation. The total analysis time from the extraction to PAH identification is approximately 35 min per sample. Limits of detection are at the sub-parts per billion levels. PAH recoveries are similar to those obtained via high-performance liquid chromatography analysis. The simplicity of the experimental procedure, the short analysis time, the selectivity, and the excellent analytical figures of merit demonstrate the advantages of this approach for routine analysis of soil samples.


The Issue of the Antiphospholipid Antibody Syndrome.
Antiphospholipid antibody syndrome (APS) is a state of hypercoagulability secondary to an autoimmune disorder. It is associated with thrombotic events in venous and arterial vessels, obstetric complications characterized by recurrent fetal losses, and increased perinatal morbidity. APS is classified as primary, when not associated with other pathologies; or secondary, when associated with an underlying autoimmune disease with, solid tumor, or hematological disorder. Clinical findings include livedo reticularis, thrombocytopenia or hemolytic anemia, maternal morbidity, and recurrent thrombotic episodes and others. Laboratory tests show circulating antiphospholipid antibodies (aPLs); however, even in the presence of these antibodies, patients can be asymptomatic. Estimates predict that about 5% of the populations have circulating aPLs, but the incidence of APS is only five cases per 100,000 people, as diagnosis of this syndrome requires clinical and laboratory findings to be simultaneously present. In cases of secondary APS, or in acute cases with imminent risk of death (as in catastrophic APS), it may be necessary to reduce aPL serum levels using immunomodulators, immunosuppressants, or plasmapheresis, in order to treat the associated pathologies. In other situations, the use of immunotherapy is not indicated. In other patients heparin, aspirin or anticoagulants either alone or associated should be administered depending on each specific case.


[Spinal cord stimulation for the treatment of refractory angina pectoris].
Spinal cord stimulation (SCS) is a well known method of treatment used worldwide since the 80's of the last century. High efficiency of SCS was observed in the treatment of various types of pain. Promising clinical results were observed in the treatment of patients with refractory angina (RA). Reduction of angina attacks reduces doses of medications and improves quality of life. SCS efficacy in RA is comparable to that observed among patients treated with cardiac surgery (CABG) and percutaneous coronary intervention (PCI), with lower risk of complications and rehospitalisation. SCS is more cost effective compared to CABG and PCI. SCS might be considered for patients who did not respond to reperfusion therapy. The mechanism of action in RA is not well understood. It is known that SCS reduces the activity of the sympathetic nervous system, by which improves the coronary perfusion and oxygenation of the heart muscle. It is important that SCS does not mask the pain related to the infarct.


Large-scale simulation of traffic flow using Markov model.
Modeling and simulating movement of vehicles in established transportation infrastructures, especially in large urban road networks is an important task. It helps in understanding and handling traffic problems, optimizing traffic regulations and adapting the traffic management in real time for unexpected disaster events. A mathematically rigorous stochastic model that can be used for traffic analysis was proposed earlier by other researchers which is based on an interplay between graph and Markov chain theories. This model provides a transition probability matrix which describes the traffic's dynamic with its unique stationary distribution of the vehicles on the road network. In this paper, a new parametrization is presented for this model by introducing the concept of two-dimensional stationary distribution which can handle the traffic's dynamic together with the vehicles' distribution. In addition, the weighted least squares estimation method is applied for estimating this new parameter matrix using trajectory data. In a case study, we apply our method on the Taxi Trajectory Prediction dataset and road network data from the OpenStreetMap project, both available publicly. To test our approach, we have implemented the proposed model in software. We have run simulations in medium and large scales and both the model and estimation procedure, based on artificial and real datasets, have been proved satisfactory and superior to the frequency based maximum likelihood method. In a real application, we have unfolded a stationary distribution on the map graph of Porto, based on the dataset. The approach described here combines techniques which, when used together to analyze traffic on large road networks, has not previously been reported.


Effects of sodium-nitroprusside and urapidil on gas exchange and ventilation-perfusion relationships in patients with congestive heart failure.
Vasodilators usually decrease arterial PO2 in patients with congestive heart failure (CHF) because of alteration in ventilation-perfusion (Va/Q) relationships. The effects of sodium nitroprusside (SNP) and urapidil (U), a new selective alpha 1-receptor antagonist, were investigated in seven patients with CHF. The distribution of ventilation and perfusion was examined using the multiple gas elimination technique. The haemodynamic responses to SNP and U were similar, cardiac index increasing by 25% with SNP and by 31% with U. Despite a similar increase of mixed venous oxygen tension, the arterial PO2 decreased from 11.3 +/- 0.8 to 9.6 +/- 0.6 kPa (p less than 0.01) with SNP but remained unchanged (11.0 +/- 0.9 vs 11.4 +/- 0.8 kPa, NS) with U. SNP and U both increased perfusion to lung units with Va/Q ratios of 0.1 or less with no change in shunt fraction. The fractional perfusion to total low Va/Q ratios (low Va/Q + shunt) was higher with SNP than with U (14.1 +/- 2.6 vs 9.5 +/- 2.3%, p less than 0.01). The results suggest that gas exchange and Va/Q relationships are altered less with U than with SNP in patients with CHF.


General practitioners' knowledge, attitudes, beliefs and practices surrounding the prescription of e-cigarettes for smoking cessation: a mixed-methods systematic review.
General practitioners (GPs) play an important role in providing patients who smoke with health information, support and treatment to encourage them to quit smoking. Despite conflicting evidence on the effectiveness of electronic cigarettes (e-cigarettes) as a smoking cessation aid, there is growing interest in the role e-cigarettes might play as an alternative to smoking tobacco. This systematic review aims to synthesise evidence from qualitative, quantitative and mixed-methods studies of the knowledge, attitudes, beliefs and social norms of GPs with respect to the use of e-cigarettes as smoking cessation aids. This study adhered to the PRISMA guidelines. Studies from MEDLINE, CINAHL, SCOPUS, PsycINFO, EMBASE and grey literature were searched. Two independent reviewers screened abstracts and full-text articles to identify studies that met the inclusion criteria. A data extraction form was used to extract relevant data from included papers and were quality appraised using the MMAT checklist. A PRISMA flow diagram was used to record the flow of papers and reasons for exclusion. Studies were included if they collected quantitative, qualitative or mixed methods data to determine knowledge, attitudes, beliefs and social norms of GPs for use of e-cigarettes as smoking cessation aids. A total of 4056 abstracts were screened and 25 articles were included. Our findings showed that GPs had mixed views on recommending e-cigarettes as a smoking cessation aid. Some GPs were optimistic and had recommended e-cigarettes to their patients. Others were reluctant and disagreed that e-cigarettes are an effective method to quit smoking. Most GPs lacked knowledge and confidence in having discussions with patients around e-cigarette safety and efficacy as smoking cessation alternatives. This systematic review shows there are mixed views on e-cigarettes as smoking cessation aids. Clear guidance on the role of e-cigarettes is needed to inform and upskill GPs about e-cigarettes for smoking cessation. CRD42021227612.


Copolymer 1 induces T cells of the T helper type 2 that crossreact with myelin basic protein and suppress experimental autoimmune encephalomyelitis.
The synthetic amino acid copolymer copolymer 1 (Cop 1) suppresses experimental autoimmune encephalomyelitis (EAE) and is beneficial in multiple sclerosis. To further understand Cop 1 suppressive activity, we studied the cytokine secretion profile of various Cop 1-induced T cell lines and clones. Unlike T cell lines induced by myelin basic protein (MBP), which secreted either T cell helper type 1 (Th1) or both Th1 and Th2 cytokines, the T cell lines/clones induced by Cop 1 showed a progressively polarized development toward the Th2 pathway, until they completely lost the ability to secrete Th1 cytokines. Our findings indicate that the polarization of the Cop 1-induced lines did not result from the immunization vehicle or the in vitro growing conditions, but rather from the tendency of Cop 1 to preferentially induce a Th2 response. The response of all of the Cop 1 specific lines/clones, which were originated in the (SJL/JxBALB/c)F1 hybrids, was restricted to the BALB/c parental haplotype. Even though the Cop 1-induced T cells had not been exposed to the autoantigen MBP, they crossreacted with MBP by secretion of interleukin (IL)-4, IL-6, and IL-10. Administration of these T cells in vivo resulted in suppression of EAE induced by whole mouse spinal cord homogenate, in which several autoantigens may be involved. Secretion of anti-inflammatory cytokines by Cop 1-induced suppressor cells, in response to either Cop 1 or MBP, may explain the therapeutic effect of Cop 1 in EAE and in multiple sclerosis.


[Some general problems of the involvement of the Department of Forensic Medical Expertise in the character formation of modern doctors].
The authors expound their views of the final goal of specialist training based at a medical higher education institution. They consider the essence of such notions as medical competence, knowledge, abilities, and skills of the future doctors. Moreover, the original approach is proposed to the organization and contents of the lectures, practical work, seminars, and self-guided work of the students seeking to acquire knowledge in forensic medicine and to the development of principles on which to base formation of clinical and expert thinking.


Assessment of zoledronic acid treatment patterns and clinical outcomes in patients with bone metastases from genitourinary cancers.
Patients with bone metastases secondary to genitourinary (GU) cancer are at risk for skeletal-related events (SREs), including bone pain requiring palliative radiotherapy, fractures or surgery to bone, spinal cord compression, and hypercalcemia of malignancy. These SREs can be debilitating and potentially life-limiting. This study examined treatment practices and the association of treatment patterns with Zometa (zoledronic acid, ZOL), an intravenous bisphosphonate (IV-BP), with SREs and fractures. (Zometa is a registered trademark of Novartis Pharmaceuticals Corporation, USA.) Retrospective analysis of commercial and Medicare Advantage enrollment and medical claims data was performed to evaluate IV-BP use and SRE patterns in adult patients with GU cancers. Criteria included diagnosis of ≥1 bone metastasis and prostate cancer (PC), renal cell carcinoma (RCC), or bladder cancer (BlC) between January 2001 and December 2006; continuous healthcare plan enrollment for ≥6 months before the index date; and no evidence of prior IV-BP use. Patients were followed until disenrollment from the healthcare plan or December 2007. Of 6347 patients (PC, n = 4976; RCC, n = 941; BlC, n = 430; mean [standard deviation] age: 68.9 [11.1] years), only approximately 23% received ZOL. The mean time between diagnosis of bone metastasis and ZOL initiation was approximately 108 days. Among patients with PC, fracture risk was significantly smaller for ZOL vs no IV-BP (incidence rate ratio = 0.70; p < 0.001), and 2-year survival was significantly longer for ZOL-treated vs no IV-BP patients (p = 0.007). Patients with longer persistency on ZOL had a smaller fracture risk than patients with shorter persistency. Sub-set analyses were not performed for RCC and BIC because the proportion of patients treated was too low. Interpretation of this claims-based analysis must be tempered by the inherent limitations of observational data, such as limited and accurate available information, and unavailable information including clinical or disease-specific parameters. Intravenous BP therapy is not always received in patients with bone metastases secondary to GU cancers, and, when used, there are typically long time periods before treatment initiation. Without IV-BPs, PC patients have significantly larger risks of fracture and death compared with ZOL-treated patients, and benefits appear to be larger with increasing persistency on ZOL.


Identification of CD4 T-cell epitopes in soluble liver antigen/liver pancreas autoantigen in autoimmune hepatitis.
Autoimmune hepatitis (AIH) is a chronic inflammatory liver disease associated with autoantibodies and liver-infiltrating lymphocytes. Although autoantibodies are tested routinely to diagnose and classify AIH, liver-infiltrating lymphocytes are regarded as the primary factor for disease pathogenesis. The purpose of this study was to identify and characterize autoantigenic peptides within human AIH-specific soluble liver antigen/liver pancreas antigen (SLA/LP) that are targeted by CD4(+) T cells and restricted by the disease susceptibility gene HLA-DRB1*0301. HLA-DRB1*0301 transgenic mice were immunized with SLA/LP. Antibody and T-cell responses were analyzed with SLA/LP-overlapping peptides in enzyme immunoassay, proliferation, and enzyme-linked immunospot (ELISpot) assays. Minimal optimal T-cell epitopes were identified, characterized with cloned T-cell hybridomas, and confirmed in tetramer and ELISpot assays with AIH patients' peripheral blood mononuclear cells. All mice developed SLA/LP-specific IgG1/IgG2a antibodies against the same SLA/LP peptides as human beings. T cells targeted several peptides within SLA/LP, 2 of which were DR3-restricted and one overlapped the sequence recognized by human autoantibodies. Minimal optimal epitopes were mapped, DRB1*0301/epitope-tetramers were generated, and the frequency and function of HLA-DRB1*0301-restricted autoantigen-specific T cells in AIH patients were analyzed with tetramer and interferon-gamma ELISpot assays. This study identified T-cell epitopes within SLA/LP, restricted by the disease susceptibility gene DRB1*0301 and in close proximity to the human autoantibody epitope. These results and the generated reagents now provide the opportunity to directly monitor autoreactive T cells in AIH patients in clinical studies.


Surgical Treatment for Empty Nose Syndrome Using Autologous Dermal Fat: Evaluation of Symptomatic Improvement.
Background: Empty nose syndrome (ENS) is caused by nasal turbinate surgery. The standard treatment for ENS is an inferior meatus augmentation procedure (IMAP) in which autologous tissue such as auricular cartilage, rib cartilage, or artificial material is transplanted into the nasal cavity. However, some challenges like a very small auricular cartilage are associated with these autologous tissue types. Moreover, since using rib cartilage is a highly invasive technique, the scar on the chest from where the harvesting is done is easily visible, and the artificial material is susceptible to infection. We used autologous dermal fat (ADF) in IMAPs in our study for the following reasons: the quantity of ADF could be increased or reduced as needed, ADF is considered a safer option than rib cartilage because it is harvested from superficial tissue, it is superior in terms of cosmetic appearance to harvested rib cartilage, and it has a lower risk of infection than any artificial material.Objective: The purpose of our study was to investigate the efficacy and safety of IMAPs using ADF.Methods: We included nine patients with ENS who underwent an IMAP using ADF. The patients' backgrounds and responses to the Empty Nose Syndrome 6-Item Questionnaire (ENS6Q) were recorded. Changes in each item of the ENS6Q before and after surgery (up to 3 months) were analyzed.Results: The postoperative ENS6Q total score and parameters were significantly better than their preoperative counterparts. Nasal dryness improved slightly less than other symptoms. There were no complications.Conclusions: The IMAP using ADF was effective in improving ENS symptoms; however, some physiological functions were difficult to improve, and dryness persisted. Autologous dermal fat is larger than auricular cartilage, less invasive than rib cartilage, and has a lower risk of infection than artificial material.


Mammographic evidence of microenvironment changes in tumorous breasts.
The microenvironment of breast tumors plays a critical role in tumorigenesis. As long as the structural integrity of the microenvironment is upheld, the tumor is suppressed. If tissue structure is lost through disruptions in the normal cell cycle, the microenvironment may act as a tumor promoter. Therefore, the properties that distinguish between healthy and tumorous tissues may not be solely in the tumor characteristics but rather in surrounding non-tumor tissue. The goal of this paper was to show preliminary evidence that tissue disruption and loss of homeostasis in breast tissue microenvironment and breast bilateral asymmetry can be quantitatively and objectively assessed from mammography via a localized, wavelet-based analysis of the whole breast. A wavelet-based multifractal formalism called the 2D Wavelet Transform Modulus Maxima (WTMM) method was used to quantitate density fluctuations from mammographic breast tissue via the Hurst exponent (H). Each entire mammogram was cut in hundreds of 360 × 360 pixel subregions in a gridding scheme of overlapping sliding windows, with each window boundary separated by 32 pixels. The 2D WTMM method was applied to each subregion individually. A data mining approach was set up to determine which metrics best discriminated between normal vs. cancer cases. These same metrics were then used, without modification, to discriminate between normal vs. benign and benign vs. cancer cases. The density fluctuations in healthy mammographic breast tissue are either monofractal anti-correlated (H < 1/2) for fatty tissue or monofractal long-range correlated (H>1/2) for dense tissue. However, tissue regions with H~1/2, as well as left vs. right breast asymetries, were found preferably in tumorous (benign or cancer) breasts vs. normal breasts, as quantified via a combination metric yielding a P-value ~ 0.0006. No metric considered showed significant differences between cancer vs. benign breasts. Since mammographic tissue regions associated with uncorrelated (H~1/2) density fluctuations were predominantly in tumorous breasts, and since the underlying physical processes associated with a H~1/2 signature are those of randomness, lack of spatial correlation, and free diffusion, it is hypothesized that this signature is also associated with tissue disruption and loss of tissue homeostasis.


Does "body talk" improve body satisfaction among same-sex couples?
Research suggests the important role of romantic partners in shaping how individuals feel about their bodies (e.g., Markey & Markey, 2013, 2014), but the processes that result in changes in body satisfaction within relationships have rarely been examined. To investigate how partners may potentially affect body image, 72 lesbian couples and 72 gay male couples (total N=288) completed body image assessments on their own and with the help of their partners. Multilevel modeling revealed that both men and women reported an improvement in body satisfaction after talking with their partners while completing the body image assessment together. Improvement in body satisfaction appeared to result from participants' reconsideration of their body ideals, not their appraisal of their current bodies. Implications of these findings for improving body image in the context of relationships are discussed.


Once-daily prolonged-release tacrolimus in de novo liver transplantation: a single center cohort study.
The feasibility of oral administration of once-daily prolonged-release tacrolimus (TAC-PR) in de novo liver transplantation is not clear and therefore was investigated further. The clinical profiles of 16 consecutive primary living donor liver transplantation (LDLT) recipients, who received oral TAC-PR once daily (TAC-PR group) between January 2009 and August 2010, were compared with those of 14 consecutive liver transplantation recipients given twice-daily tacrolimus (TAC; TAC group) between August 2006 and January 2009. Of the 14 patients in the TAC group, 9 received LDLT (TAC-L subgroup). Patient characteristics were similar between groups. Trough levels of TAC during the first 3 months after liver transplantation were well-adjusted in both groups. Dose adjustment was more frequently required (31.3%) in the TAC-PR group and the total amount of TAC was significantly higher in the TAC-PR group (181.1±75.3mg) than in the TAC-L group (100.2±53.8mg, p=0.014). The incidence of biopsy-proven acute cellular rejection, renal dysfunction, other morbidities and hospital stay length were similar between groups. Oral administration of TAC-PR for de novo liver transplantation recipients was well tolerated with similar safety and efficacy profiles as traditional twice-daily TAC with closely controlled adjustment of the TAC-PR dose.


Spatially controlled co-culture of neurons and glial cells.
The ability to create and maintain neuron and glial cell co-cultures is important for neuronal regeneration as well as for fundamental studies on neuron and glial cell interactions. We demonstrate here a method for spatially controlling the arrangement of neurons and glial cells. Line patterns of cell resistant, poly(oligoethyleneglycol methacrylate-co-methacrylic acid), was microcontact printed on various substrates to spatially control the attachment of neurons. Neuron-like cells, PC12 and SH-SY5Y cells, were confined within the unprinted line patterns and extended neurites along the line patterns. Subsequent attachment of glial cells was accomplished by converting the originally cell-resistant line patterns of poly(oligoethyleneglycol methacrylate-co-methacrylic acid) to cell adhesive by electrostatic adsorption of cationic poly-lysine, chitosan, or poly(ethyleneimine). This method for creating patterned co-cultures of neuron and glial cells provides a useful tool for investigating neuron-glial cell interactions and has potential applications in the repair or regeneration of nervous systems.


Retrospective Comparison of Survival Projections for CAR T-Cell Therapies in Large B-Cell Lymphoma.
Durable remission has been observed in patients with relapsed or refractory (R/R) large B-cell lymphoma (LBCL) treated with chimeric antigen receptor (CAR) T-cell therapy. Consequently, hazard functions for overall survival (OS) are often complex, requiring the use of flexible methods for extrapolations. We aimed to retrospectively compare the predictive accuracy of different survival extrapolation methods and evaluate the validity of goodness-of-fit (GOF) criteria-based model selection for CAR T-cell therapies in R/R LBCL. OS data were sourced from JULIET, ZUMA-1, and TRANSCEND NHL 001. Standard parametric, mixture cure, cubic spline, and mixture models were fit to multiple database locks (DBLs), with varying follow-up durations. GOF was assessed using the Akaike information criterion and Bayesian information criterion. Predictive accuracy was calculated as the mean absolute error (MAE) relative to OS observed in the most mature DBL. For all studies, mixture cure and cubic spline models provided the best predictive accuracy for the least mature DBL (MAE 0.013‒0.085 and 0.014‒0.128, respectively). The predictive accuracy of the standard parametric and mixture models showed larger variation (MAE 0.024‒0.162 and 0.013‒0.176, respectively). With increasing data maturity, the predictive accuracy of standard parametric models remained poor. Correlation between GOF criteria and predictive accuracy was low, particularly for the least mature DBL. Our analyses demonstrated that mixture cure and cubic spline models provide the most accurate survival extrapolations of CAR T-cell therapies in LBCL. Furthermore, GOF should not be the only criteria used when selecting the optimal survival model.


A novel human cytomegalovirus glycoprotein, gpUS9, which promotes cell-to-cell spread in polarized epithelial cells, colocalizes with the cytoskeletal proteins E-cadherin and F-actin.
Processes by which human herpesviruses penetrate and are released from polarized epithelial cells, which have distinct apical and basolateral membrane domains differing in protein and lipid content, are poorly understood. We recently reported that human cytomegalovirus (CMV) mutants with deletions of the gene US9 formed wild-type plaques in cultures of human fibroblasts but were impaired in the capacity for cell-to-cell spread in polarized human retinal pigment epithelial cells. Unlike the glycoproteins that are required for infection, the protein encoded by CMV US9 plays an accessory role by promoting dissemination of virus across cell-cell junctions of polarized epithelial cells. To identify the product and investigate its specialized functions, we selected Madine-Darby canine kidney II (MDCK) epithelial cells that constitutively express CMV US9 or, as a control, US8. The gene products, designated gpUS9 and gpUS8, were glycosylated proteins of comparable molecular masses but differed considerably in intracellular distribution and solubility. Immunofluorescence laser scanning confocal microscopy indicated that, like gpUS8, gpUS9 was present in the endoplasmic reticulum and Golgi compartments of nonpolarized cells. In polarized epithelial cells, gpUS9 also accumulated along lateral membranes, colocalizing with cadherin and actin, and was insoluble in Triton X-100, a property shared with proteins that associate with the cytoskeleton. We hypothesize that gpUS9 may enhance the dissemination of CMV in infected epithelial tissues by associating with the cytoskeletal matrix.


Video-speed detection of the absolute position of a light point on a large-area photodetector based on luminescent waveguides.
A large-area photosensor is presented that uses the coupling of light into the planar waveguide mode of a polycarbonate foil by luminescent dyes to extend the active area of silicon photodiodes attached to the surface of the foil in a regular pattern. The photodiode signal is directly related to the distance between the point where light is coupled into the foil and the photodiode, thus enabling a precise recovery of the position of a localized light signal hitting the foil. We present a large-area device that can trace the movement of a light point generated by a laser pointer hitting its surface.


WiFi RFID demonstration for resource tracking in a statewide disaster drill.
To investigate the capabilities of Radio Frequency Identification (RFID) tracking of patients and medical equipment during a simulated disaster response scenario. RFID infrastructure was deployed at two small rural hospitals, in one large academic medical center and in two vehicles. Several item types from the mutual aid equipment list were selected for tracking during the demonstration. A central database server was installed at the UC Davis Medical Center (UCDMC) that collected RFID information from all constituent sites. The system was tested during a statewide disaster drill. During the drill, volunteers at UCDMC were selected to locate assets using the traditional method of locating resources and then using the RFID system. This study demonstrated the effectiveness of RFID infrastructure in real-time resource identification and tracking. Volunteers at UCDMC were able to locate assets substantially faster using RFID, demonstrating that real-time geolocation can be substantially more efficient and accurate than traditional manual methods. A mobile, Global Positioning System (GPS)-enabled RFID system was installed in a pediatric ambulance and connected to the central RFID database via secure cellular communication. This system is unique in that it provides for seamless region-wide tracking that adaptively uses and seamlessly integrates both outdoor cellular-based mobile tracking and indoor WiFi-based tracking. RFID tracking can provide a real-time picture of the medical situation across medical facilities and other critical locations, leading to a more coordinated deployment of resources. The RFID system deployed during this study demonstrated the potential to improve the ability to locate and track victims, healthcare professionals, and medical equipment during a region-wide disaster.


A processed multidomain mycoplasma hyopneumoniae adhesin binds fibronectin, plasminogen, and swine respiratory cilia.
Porcine enzootic pneumonia is a chronic respiratory disease that affects swine. The etiological agent of the disease, Mycoplasma hyopneumoniae, is a bacterium that adheres to cilia of the swine respiratory tract, resulting in loss of cilia and epithelial cell damage. A M. hyopneumoniae protein P116, encoded by mhp108, was investigated as a potential adhesin. Examination of P116 expression using proteomic analyses observed P116 as a full-length protein and also as fragments, ranging from 17 to 70 kDa in size. A variety of pathogenic bacterial species have been shown to bind the extracellular matrix component fibronectin as an adherence mechanism. M. hyopneumoniae cells were found to bind fibronectin in a dose-dependent and saturable manner. Surface plasmon resonance was used to show that a recombinant C-terminal domain of P116 bound fibronectin at physiologically relevant concentrations (K(D) 24 ± 6 nm). Plasmin(ogen)-binding proteins are also expressed by many bacterial pathogens, facilitating extracellular matrix degradation. M. hyopneumoniae cells were found to also bind plasminogen in a dose-dependent and saturable manner; the C-terminal domain of P116 binds to plasminogen (K(D) 44 ± 5 nm). Plasminogen binding was abolished when the C-terminal lysine of P116 was deleted, implicating this residue as part of the plasminogen binding site. P116 fragments adhere to the PK15 porcine kidney epithelial-like cell line and swine respiratory cilia. Collectively these data suggest that P116 is an important adhesin and virulence factor of M. hyopneumoniae.


Histone deacetylase inhibitors and cell proliferation in pea root meristems.
The histone deacetylase (HDA) inhibitors, trichostatin A (TSA) and HC toxin halt mitosis in cultured root meristems of Pisnum sativum, while the anti-protozoan HDA inhibitor apicidin is ineffective. Two-dimensional PAGE of proteins from root meristems exposed to TSA and HC toxin did not show significant differences compared to controls, although a previously tested HDA inhibitor, butyrate, exhibited dramatic variations in its protein profile. Northern analysis of butyrate- and TSA-treated root meristems indicated that non-proliferating cells are expressing significant amounts of transcripts of the known cell proliferation associated genes: histone H2A, MAP kinase, cycA2:1 and cdc2. Western analysis reveals the presence of hyperacetylated nuclear proteins in HDA-inhibitor treated cells. These results suggest that the HDA inhibitors, butyrate and TSA, halt mitosis without down-regulating genes that typically have low or nonexistent expression levels in non-dividing cells.


Emergency ultrasound of the abdominal aorta by UK emergency physicians: a prospective cohort study.
To establish whether UK emergency physicians could reliably perform focused ultrasound of the abdominal aorta in patients with suspected abdominal aortic aneurysm (AAA). A prospective cohort study was conducted in the emergency department of a tertiary level UK teaching hospital. All patients who underwent an abdominal aortic ultrasound by an emergency physician during a 12 month period from January to December 2005 were included. The principle outcomes were presence of an AAA (external wall diameter >3 cm) or death from ruptured AAA. Outcome data were obtained from paper and electronic patient records and primary care telephone follow up. 120 focused ultrasound scans looking for AAA were performed by 19 different UK emergency physicians of various grades. Of the 120 scans, 26 (22%) were positive for an AAA, of which 17 cases represented a new diagnosis. Ruptured aneurysms represented 46% (12/26) of all positive scans, of which four patients underwent emergency repair. In the remaining 14 patients the AAA was an incidental finding that was not the reason for their presentation to the emergency department. Emergency ultrasound had a sensitivity of 96.3% (95% confidence interval (CI) 81.0% to 99.9%); a specificity of 100% (95% CI 91.8% to 100%); a negative predictive value of 98.6% (95% CI 88.0% to 99.9%); and positive predictive value of 100% (95% CI 86.8% to 100%) for the detection of AAA. Emergency ultrasound scanning by UK emergency physicians has high sensitivity and specificity for identifying AAA, consistent with international experience.


Auditory perceptions induced by low-frequency acoustic and electric stimulation.
A comparison was made between listeners' perceptions of low frequency (LF) acoustic and electric stimuli. One normal-hearing S participated, and 3 profoundly deaf pts with intracochlear electrode systems. All Ss described their auditory perceptions to pulsatile and sinusoidal stimuli from 5 to 60 c/s. LF acoustic and electric stimuli produced similar auditory perceptions; no tactual component was found to electrical stimuli, although tactual components have been reported for LF acoustic stimuli; and at 5 c/s perceptual differences were noted between pulsed and sinusoidal stimuli, differences believed to be related to the degree of neural synchrony elicited by the two types of stimulus. This study provides direct support for the theory that in normal listeners LF auditory perception is attributable to a neural volley mechanism.


Association Between Living Kidney Donor Postdonation Hypertension and Recipient Graft Failure.
Recipients of kidneys from living donors who subsequently develop end-stage renal disease (ESRD) also have higher graft failure, suggesting the 2 donor kidneys share risk factors that could inform recipient outcomes. Given that donor ESRD is rare, an earlier and more common postdonation outcome could serve as a surrogate to individualize counseling and management for recipients. Hypertension is a frequent event before donor ESRD; thus, early postdonation hypertension might indicate higher risk of graft failure. We studied Scientific Registry of Transplant Recipients data to quantify the association between early postdonation hypertension and recipient graft failure using propensity score-weighted Cox proportional hazards regression. We also examined the association between postdonation systolic blood pressure and graft failure. Of 37 901 recipients, 2.4% had a donor who developed hypertension within 2 years postdonation. Controlling for donor and recipient characteristics, recipients whose donors developed hypertension had no higher risk for graft failure (adjusted hazard ratio [aHR] 1.03, 95% confidence interval [CI] 0.85-1.25, P = 0.72). This was consistent among subgroups of recipients at higher risk for adverse outcomes due to hyperfiltration: African American recipients (aHR 1.10, 95% CI 0.70-1.73, P = 0.68) and those with ESRD caused by hypertension (aHR 1.10, 95% CI 0.65-1.85, P = 0.73) or diabetes (aHR 0.80, 95% CI 0.56-1.13, P = 0.20). However, graft failure was associated with postdonation systolic blood pressure (per 10 mm Hg, aHR 1.05, 95% CI 1.03-1.08, P < 0.001). Although postdonation systolic blood pressure is associated with graft failure, the reported diagnosis of hypertension as determined by the requirement for blood pressure treatment early postdonation did not portend a higher risk of recipient graft failure in the same way as eventual postdonation ESRD.


Effect of dance labor on the management of active phase labor pain & clients' satisfaction: a randomized controlled trial study.
There are a wide variety of non- pharmacologic pain relief techniques for labor which include pelvic movement, upright position, back massage and partner support during the first stage of labor. The effectiveness of dance labor- which is a combination of these techniques- has not been evaluated. This study aimed to evaluate the effectiveness of dance labor in pain reduction and woman's satisfaction during the first stage of labor. 60 primiparous women aged 18-35 years old were randomly assigned to dance labor and control groups. In the dance labor group, women were instructed to do standing upright with pelvic tilt and rock their hips back and forth or around in a circle while their partner massaged their back and sacrum for a minimum of 30 minutes. In the control group, the participants received usual care during physiologic labor. Pain and satisfaction scores were measured by Visual Analogue Scale. Data were analyzed by using the t. test and Chi-square. Mean pain score in the dance labor group was significantly lower than the control group (P < 0.05). The mean satisfaction score in the dance labor group was significantly higher than in the control group (P < 0.05). Dance labor which is a complementary treatment with low risk can reduce the intensity of pain and increase mothers, satisfaction with care during the active phase of labor.


Self-assembly and pH response of electroactive liquid core-tetra(aniline) shell microcapsules.
We developed a simple one-pot method to fabricate electroactive tetra(aniline)-based microcapsules, which exhibit a wide range of pH response by self-assembly. In this procedure, the microcapsules are prepared by the facile dialysis-induced self-assembly of tetra(aniline), TANI, doped with acetic acid (HAc), onto the oil-in-water droplet interface. The chemical structure of the shell is characterized by FT-IR and UV-Vis spectroscopy. The electrochemical properties, self-assembly and pH response of microcapsule dispersions are further investigated by cyclic voltammetry, optical and electron microscopy, dynamic light scattering and aqueous electrophoresis. The size of the microcapsules increases gradually from 1.25 μm to 2.93 μm when the concentration of HAc changes from 12 M to 14 M and higher (to become glacial acetic acid). Furthermore, we show that aggregation and stability of the microcapsules can be controlled through changes in pH. This strategy appears to be a general scheme for producing other oligo(aniline)-based microcapsules.


Bone marrow fibrosis in childhood acute lymphoblastic leukaemia.
Bone marrow trephine biopsy specimens were obtained at diagnosis from 63 of 76 consecutively presenting children with acute lymphoblastic leukaemia (ALL). The association between marrow fibrosis and presenting features, including immunophenotype, was analysed. Reticulin was increased in 45 of 56 cases in which blasts expressed B lineage markers, but in only one of seven with T-ALL. A weak association was also found between marrow fibrosis and splenomegaly in those with common ALL. Marrow fibrosis is apparently associated with some examples of ALL of B cell lineage, but precisely which subtypes and whether the phenomenon is clinically important remain to be determined.


Disorders of cranial nerves IX and X.
The glossopharyngeal and vagus nerves mediate the complex interplay between the many functions of the upper aerodigestive tract. Defects may occur anywhere from the brainstem to the peripheral nerve and can result in significant impairment in speech, swallowing, and breathing. Multiple etiologies can produce symptoms. In this review, the authors broadly examine the normal functions, clinical examination, and various pathologies of cranial nerves IX and X.


Use and Evaluation of a Cooling Aid in Laser-Assisted Dental Surgery: An Innovative Study.
The aim of this study was to evaluate the beneficial effect of NeoHeal during laser-assisted oral surgery. Laser-assisted surgery is a less invasive technique than conventional surgery using a blade; however, the heat produced by the laser can damage tissue proximal to the treated area. We hypothesized the hydrogel NeoHeal could be applied during oral surgery and have similar benefits as those already described for dermatological surgery. In our randomized controlled trial, we first selected 30 patients who were to undergo laser surgical intervention using a diode laser. Both at the end of intervention and at follow-up, each patient filled out a questionnaire comprising two questions. The answers to the questions were statistically analyzed with regard to frequency of each response, and validated by the Wilcoxon test with a p value <0.05 being considered significant. The use of NeoHeal significantly reduced the pain experienced by patients both during the intervention and at the 7-day follow-up. NeoHeal clearly reduces surgery-related pain experienced by patients. The results show positive feedback from the patients, demonstrating how the hydrogel pads are necessary, useful, and effective as a cooling aid in laser-assisted dental surgery.


Neurosteroid modulation of embryonic neuronal survival in vitro following anoxia.
Neurosteroids are synthesized de novo in the brain from cholesterol or peripheral steroid precursors and modulate inhibitory gamma-aminobutyric acid (GABA(A)) and excitatory N-methyl-D-aspartate (NMDA) receptors. Evidence indicates that neurosteroids are neuroprotective and important during neurodevelopment. We tested the hypothesis that neurosteroids increase embryonic neuronal survival following anoxia in rat embryonic day 18 cerebral cortical cultures to examine potential neurosteroid modulation of this insult during early development. Twenty-four hours after plating in serum-free medium, cultures were exposed to DHEA, DHEAS, or allopregnanolone (10(-10), 10(-8), or 10(-6) M), or vehicle, for 24 h (n=9 per treatment condition). Cultures were then subjected to anoxia for 2 h and subsequently reincubated for 24 h prior to neuron immunostaining with microtubule-associated protein 2 (MAP-2) antibody. Supernatant from DHEA and DHEAS-exposed cultures was tested for 17beta-estradiol metabolite formation by radioimmunoassay. DHEA 10(-6) and 10(-8) M significantly increased neuron survival by 85-87% following anoxia. DHEAS 10(-6) M significantly increased neuron survival by 74% following anoxia, but DHEAS 10(-10) M decreased neuron survival after this insult. Allopregnanolone had modest effects on neuron survival that did not attain statistical significance. 17beta-Estradiol concentrations were below the limit of detection in all specimens tested (sensitivity 4.7 nM). Our data indicate that pretreatment with DHEA and DHEAS at physiologically relevant concentrations promotes neuronal survival following anoxia in embryonic rat cerebral cortical cultures, and that these effects are not secondary to 17beta-estradiol metabolite formation. DHEA and DHEAS modulation of anoxia in embryonic neurons may be relevant to disorders of neurodevelopment involving this insult.


Oncolymphology: Immune Interactions and Cancer.
The proposed term "oncolymphology" encompasses the intimate relationship between cancer growth and the immune responses.


First-line treatment in the management of advanced renal cell carcinoma: systematic review and network meta-analysis.
To conduct a systematic review and network meta-analysis (NMA) to assess effectiveness of first-line treatments for advanced renal cell carcinoma (RCC). Database searches were conducted to identify randomized controlled trials (RCTs) reporting results for eligible treatments. A fixed-effect Bayesian NMA was conducted to assess the relative effectiveness of treatments, with progression-free survival (PFS) reported as hazard ratios (HRs) and 95% credible intervals (CrIs). Eleven unique RCTs were suitable for inclusion in the NMA. In the base case, in terms of PFS, sunitinib was superior compared with bevacizumab + IFN-α (HR = 0.79, 95% CrI: 0.64 - 0.96), everolimus (HR = 0.70, 95% CrI: 0.56 - 0.87), sorafenib (HR = 0.56, 95% CrI: 0.40 - 0.77) and temsirolimus + bevacizumab (HR = 0.74, 95% CrI: 0.56 - 0.96). Although, the point values for the mean and median HRs were < 1.0, there was no significant difference in PFS between sunitinib and axitinib, pazopanib or tivozanib. Although sensitivity analyses impacted the results of the NMA, no treatment was significantly more efficacious than sunitinib. Results from this analysis suggest that there is no treatment superior to the current benchmark treatment, sunitinib, in the management of advanced RCC in the first-line setting.


[Malignant neuroleptic syndrome and malignant hyperthermia--a comparison].
Malignant Hyperthermia is a specific potentially fatal condition which occurs in susceptible individuals in response to various triggering mechanisms, of which anesthetic agents have been found to be the most common offenders. Leading symptoms are generalized muscular rigidity and hyperpyrexia. The etiology seems to be associated with some inherited disturbance in muscle metabolism related to calcium regulation. This syndrome is known for 25 years, the mechanism of triggering, the genetics and the treatment have been able to get examined by animal model as malignant hyperthermia syndrome may occur in swine as well. Pharmacological in-vitro studies on biopsy specimen of muscle fragments are presently one of the most accepted means for pre-anesthetic diagnosis, the hydantoin derivate dantrolene sodium is the only known specific drug in treatment and prophylaxis. Neuroleptic Malignant Syndrome which occurs in patients treated with neuroleptics shows almost identical symptoms. Although the pathogenesis is still unknown, most authors believe a neuroleptic-induced alteration of central neuroregulatory mechanisms is involved. Alternative etiologic mechanisms are suggested by the striking similarities noted between the Neuroleptic Malignant Syndrome and Malignant Hyperthermia. Both disorders might be based on common pathophysiologic mechanisms and might be induced in susceptible patients by a variety of pharmacologic agents. The authors present and compare the actual knowledge concerning symptoms, course and therapy of both syndromes on the basis of 126 case studies of the Neuroleptic Malignant Syndrome from literature.


Changes in the distribution of multispecies pest assemblages affect levels of crop damage in warming tropical Andes.
Climate induced species range shifts might create novel interactions among species that may outweigh direct climatic effects. In an agricultural context, climate change might alter the intensity of competition or facilitation interactions among pests with, potentially, negative consequences on the levels of damage to crop. This could threaten the productivity of agricultural systems and have negative impacts on food security, but has yet been poorly considered in studies. In this contribution, we constructed and evaluated process-based species distribution models for three invasive potato pests in the Tropical Andean Region. These three species have been found to co-occur and interact within the same potato tuber, causing different levels of damage to crop. Our models allowed us to predict the current and future distribution of the species and therefore, to assess how damage to crop might change in the future due to novel interactions. In general, our study revealed the main challenges related to distribution modeling of invasive pests in highly heterogeneous regions. It yielded different results for the three species, both in terms of accuracy and distribution, with one species surviving best at lower altitudes and the other two performing better at higher altitudes. As to future distributions our results suggested that the three species will show different responses to climate change, with one of them expanding to higher altitudes, another contracting its range and the other shifting its distribution to higher altitudes. These changes will result in novel areas of co-occurrence and hence, interactions of the pests, which will cause different levels of damage to crop. Combining population dynamics and species distribution models that incorporate interspecific trade-off relationships in different environments revealed a powerful approach to provide predictions about the response of an assemblage of interacting species to future environmental changes and their impact on process rates.


When is an Offset Not an Offset? A Framework of Necessary Conditions for Biodiversity Offsets.
Biodiversity offsets have become a widely accepted means of attempting to compensate for biodiversity loss from development, and are applied in planning and decision-making processes at many levels. Yet their use is contentious, and numerous problems with both the concept and the practice have been identified in the literature. Our starting point is the understanding that offsets are a kind of biodiversity compensation measure through which the goal of no net loss (or net gain) of biodiversity can be at least theoretically achieved. Based on a typology of compensation measures distinguishing between habitat protection, improvement (including restoration, habitat creation and improved management practices) and other compensation, we review the literature to develop a framework of conditions that must be met if habitat protection and improvement initiatives can be truly considered offsets and not merely a lesser form of compensation. It is important that such conceptual clarity is reflected in offset policy and guidance, if offsets are to be appropriately applied and to have any chance of fully compensating for biodiversity loss. Our framework can be used to support the review and ongoing development of biodiversity offset policy and guidance, with the aim of improving clarity, rigour and therefore the chances that good biodiversity outcomes can be achieved.


Diagnosis of an "isodense" pituitary microadenoma by dynamic CT scanning. Case report.
A case of Nelson's syndrome with an adrenocorticotropic hormone-secreting pituitary chromophobe microadenoma is presented to demonstrate the potential capability of rapid sequential (dynamic) computerized tomography (CT) scanning for the diagnosis of a pituitary microadenoma that was isodense with the adjacent pituitary gland on conventional enhanced CT scanning. The dynamic CT scans showed transient high density in this microadenoma contrasting with the pituitary gland in the early-enhancement phase, and thereafter the contrast density was indistinguishable from that of the pituitary gland in the delayed-enhancement phase. For the detection of pituitary microadenoma, dynamic CT combined with subsequent delayed CT scanning can provide diagnostic and localizing information.


Interspecies microbiome transplantation recapitulates microbial acquisition in mosquitoes.
Mosquitoes harbor microbial communities that play important roles in their growth, survival, reproduction, and ability to transmit human pathogens. Microbiome transplantation approaches are often used to study host-microbe interactions and identify microbial taxa and assemblages associated with health or disease. However, no such approaches have been developed to manipulate the microbiota of mosquitoes. Here, we developed an approach to transfer entire microbial communities between mosquito cohorts. We undertook transfers between (Culex quinquefasciatus to Aedes aegypti) and within (Ae. aegypti to Ae. aegypti) species to validate the approach and determine the number of mosquitoes required to prepare donor microbiota. After the transfer, we monitored mosquito development and microbiota dynamics throughout the life cycle. Typical holometabolous lifestyle-related microbiota structures were observed, with higher dynamics of microbial structures in larval stages, including the larval water, and less diversity in adults. Microbiota diversity in recipient adults was also more similar to the microbiota diversity in donor adults. This study provides the first evidence for successful microbiome transplantation in mosquitoes. Our results highlight the value of such methods for studying mosquito-microbe interactions and lay the foundation for future studies to elucidate the factors underlying microbiota acquisition, assembly, and function in mosquitoes under controlled conditions. Video Abstract.


Hemorrhagic enteritis of turkeys: electron microscopy of the causal virus.
Spleens of turkey poults inoculated with hemorrhagic enteritis virus were examined by electron microscopy. Negatively stained infectious extracts of spleen tissue contained adenolike virions averaging 75-80 nm in diameter. Intranuclear virions, either scattered or in crystalline arrays, were observed in thin sections of spleen tissue four days after inoculation. The chromatin was condensed and marginated in the nuclei of most cells which contained virions. Some virions in sections appeared as hollow cores with a double-layered capsid. The outer capsid was 75-80 nm in diameter, and the inner capsid 35-40 nm. Virions were not observed in cells or extracts of spleens from uninoculated poults. The detection of intranuclear virions at the time when symptoms appeared and specific antigen titer increased suggests that the adeno-like virions are the causal agent of hemorrhagic enteritis of turkeys.


A neutralizable epitope common to the envelope glycoproteins of ecotropic, polytropic, xenotropic, and amphotropic murine leukemia viruses.
An epitope common to all classes of murine leukemia viruses (MuLVs) was detected by reactivity of MuLVs with a rat monoclonal antibody (MAb) termed 83A25. The antibody is of the immunoglobulin G2a isotype and was derived after fusion of NS-1 myeloma cells with spleen cells from a Fischer rat immunized with a Friend polytropic MuLV. The antibody reacted with nearly all members of the ecotropic, polytropic, xenotropic, and amphotropic classes of MuLVs. Unreactive viruses were limited to the Friend ecotropic MuLV, Rauscher MuLV, and certain recombinant derivatives of Friend ecotropic MuLV. The presence of an epitope common to nearly all MuLVs facilitated a direct quantitative focal immunofluorescence assay for MuLVs, including the amphotropic MuLVs for which no direct assay has been previously available. Previously described MAbs which react with all classes of MuLVs have been limited to those which react with virion core or transmembrane proteins. In contrast, protein immunoblot and immunoprecipitation analyses established that the epitope reactive with MAb 83A25 resides in the envelope glycoproteins of the viruses. Structural comparisons of reactive and nonreactive Friend polytropic viruses localized the epitope near the carboxyl terminus of the glycoprotein. The epitope served as a target for neutralization of all classes of MuLV with MAb 83A25. The efficiency of neutralization varied with different MuLV isolates but did not correlate with MuLV interference groups.


Human Exposure to Potentially Toxic Elements from the Consumption of Soybean Beverages Commercialized in Spain.
Plant-based beverages have been positioned as an alternative to cow's milk. Some of the most commonly consumed plant-based beverages are those made from soybeans. Soybean beverages can be an important source of essential elements. However, soybean plants can be exposed to high concentrations of contaminants, such as toxic metals, from the soil and the environment. The objective of this study was to determine the concentrations of potentially toxic elements (PTEs; Fe, Cu, Cr, Co, Mn, Mo, Zn, B, Ba, Ni, Li, Sr, V, Al, Cd, and Pb) to evaluate the human exposure and toxic risk from the consumption of these soybean beverages. Forty samples of various brands of soybean beverages were analyzed by inductively coupled plasma-optical emission spectroscopy. The most abundant PTE was Al (3.93 mg/L), followed by Fe (1.91 mg/L) and Ba (0.69 mg/L). Based on a mean consumption of two servings of soybean beverage per day (250 mL per serving), this consumption results in a daily intake of Al and Pb that is significant but still below of the maximum recommended amount. However, higher consumption could exceed the maximum recommended Al and Pb values, with consequent risks to the human health.


Platelet and vessel wall interaction and the genesis of atherosclerosis.
On the basis of in vitro studies, biochemistry and cell biology the interactions between platelets, other circulating blood cells and the vessel wall are complex and wide-ranging. In essence, the effects of platelets adhering to injured luminal surface might include: Aggregation and adherence of further platelets; Activation of coagulation and fibrin formation; Displacement of AT-III from endothelial surface; Recruitment and activation of potentially injurious polymorphonuclear leukocytes and monocytes; Recruitment and proliferation of smooth muscles cells from the vessel media to the intima; Vasoconstriction; Initiation of feedback mechanisms such as PGI2 release to limit the process. These effects are capable of contributing to the initial lesions and the progression of the atherosclerosis. As the lesions become more complex, and particularly as the normal endothelium is lost, there is morphological evidence that there is deposition of thrombus (for discussion see Chapter 4) and 111In-labelled platelet imaging experiments in patients document platelet uptake onto atherosclerotic plaques (Davis et al, 1978).


DNA oxidation induced by cyclooxygenase-2.
The inducible form of cyclooxygenase, COX-2, has been shown to be overexpressed in various types of tumors, including colon and prostate cancer. Several studies indicate that COX-2 inhibition can be beneficial for the prevention of these types of cancer. Since COX-2 reactions involve production of reactive oxygen radicals that can potentially damage biological macromolecules, we explored the possibility that DNA and/or nucleosides can be oxidized during cyclooxygenase reactions. When DNA or nucleosides were incubated with COX-2 and arachidonic acid, a significant increase in the amount of 8-oxo-2'-deoxyguanosine was observed. This increase was enzyme-dependent and could be prevented by COX-2 inhibitors as well as by antioxidants. These data indicate that peroxyl radicals or other oxidized species formed during conversion of arachidonic acid to prostaglandin G(2) might be responsible for the observed oxidation. These results suggest also that overexpression of COX-2 in inflammatory diseases places an additional burden on antioxidative defenses of the cell, which might contribute to DNA oxidation and the induction of mutations.


Pyrophosphate heart scan in patients with progressive systemic sclerosis.
Scintigraphic examination of the myocardium, using 99mTc-labelled pyrophosphate, was carried out in 17 patients suffering from systemic sclerosis. This connective tissue disorder very often affects the myocardium secondarily. The results of the cardiac scan were compared with the information obtained from the electrocardiogram of systolic time intervals. In addition, spirometry was undertaken to detect a potential relation between cardiac and pulmonary involvement. The scan was found to be positive in seven patients and electrocardiographic findings were pathological in five patients only. The systolic time intervals were abnormal in three patients only. A ventilation disturbance was recorded in 10 patients. No clear relation was found between the results of the individual examinations. It is concluded that pyrophosphate heart scintigraphy may detect myocardial impairment in some cases of systemic sclerosis before it is manifested by heart failure. Examination of systolic time intervals is of little importance.


Changes in differential gene expression during a fatal stroke.
We present a young woman (with an identical twin sister) who arrived at the Emergency Department (ED) within 1 hour of her initial stroke symptoms. Previous microarray studies have demonstrated differential expression of multiple genes between stroke patients and healthy controls. However, for many of these studies there is a significant delay between the initial symptoms and collection of blood samples, potentially leaving the important early activators/regulators of the inflammatory response unrecognised. Blood samples were collected from the patient for an analysis of differential gene expression over time during the evolution of a fatal stroke. The time points for blood collection were ED arrival (T0) and 1, 3 and 24 hours post ED arrival (T1, T3 and T24). This was compared to her identical twin and an additional two age and sex-matched healthy controls. When compared to the controls, the patient had 12 mRNA that were significantly upregulated at T0, and no downregulated mRNA (with a cut off fold change value ±1.5). Of the 12 upregulated mRNA at T0, granzyme B demonstrated the most marked upregulation on arrival, with expression steadily declining over time, whereas S100 calcium-binding protein A12 (S100A12) gene expression increased from T0 to T24, remaining >two-fold above that in the healthy controls at T24. Other genes, such as matrix metalloproteinase 9, high mobility group box 2 and interleukin-18 receptor I were not upregulated at T0, but they demonstrated clear upregulation from T1–T3, with gene expression declining by T24. A greater understanding of the underlying immunopathological mechanisms that are involved during the evolution of ischaemic stroke may help to distinguish between patients with stroke and stroke mimics.


Expert and Novice Performers Respond Differently to Attentional Focus Cues for Speed Jump Roping.
Directing attention to an external focus has been shown to facilitate motor performance. For expert performers, however, results have been mixed. Additionally, little is currently known about how focus cues affect the performance of complex continuous whole-body coordination tasks involving object manipulation such as jump roping. The purpose of this study was to examine the effects of attentional focus cues on single-rope speed jumping by experts and novices. The cues directed attention toward the upper (UP) or lower (LB) body and either internally (IN) or externally (EX). Participants (N = 30) completed bouts of speed jumping during a baseline trial and under experimental conditions (UPIN, UPEX, LBIN, LBEX). Jumps and errors were recorded for each trial. Number of jumps (NJ) and errors (NE) were analyzed using separate Friedman's Tests comparing for each group to compare trials, with Wilcoxon Signed-Rank Tests for post hoc comparisons. Cumulative number of jumps (CNJ) and errors (CNE) for each condition were compared using separate Friedman's Tests with Wilcoxon Signed-Rank Tests for post hoc comparisons. For experts, baseline NJ was significantly higher than NJ for each trial under the UPIN, UPEX, and LBIN conditions. No differences between baseline NE and any trials were detected. Additionally, no differences were detected between conditions for NJ or NE. For novices, baseline NJ and NE were significantly higher and lower, respectively, compared to Trial 1 under the LBEX condition. Both the UPIN and UPEX conditions produced higher CNJ and lower CNE than the LBIN and LBEX conditions, respectively. Results showed that experts and novices responded in distinctly different patterns to the four conditions. Experts showed degraded performance under the UPIN, UPEX, and LBIN conditions whereas novices only showed temporarily degraded performance under the LBEX condition. These findings may reflect differences in mastery of whole-body coordination and are partially consistent with the Constrained Action Hypothesis (CAH) despite not supporting specific predictions related to the benefit of external focus cues.


Targeted Detection of SARS-CoV-2 Nucleocapsid Sequence Variants by Mass Spectrometric Analysis of Tryptic Peptides.
COVID-19 vaccines are becoming more widely available, but accurate and rapid testing remains a crucial tool for slowing the spread of the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) virus. Although the quantitative reverse transcription-polymerase chain reaction (qRT-PCR) remains the most prevalent testing methodology, numerous tests have been developed that are predicated on detection of the SARS-CoV-2 nucleocapsid protein, including liquid chromatography-tandem mass spectrometry (LC-MS/MS) and immunoassay-based approaches. The continuing emergence of SARS-CoV-2 variants has complicated these approaches, as both qRT-PCR and antigen detection methods can be prone to missing viral variants. In this study, we describe several COVID-19 cases where we were unable to detect the expected peptide targets from clinical nasopharyngeal swabs. Whole genome sequencing revealed that single nucleotide polymorphisms in the gene encoding the viral nucleocapsid protein led to sequence variants that were not monitored in the targeted assay. Minor modifications to the LC-MS/MS method ensured detection of the variants of the target peptide. Additional nucleocapsid variants could be detected by performing the bottom-up proteomic analysis of whole viral genome-sequenced samples. This study demonstrates the importance of considering variants of SARS-CoV-2 in the assay design and highlights the flexibility of mass spectrometry-based approaches to detect variants as they evolve.


Human T cell leukemia virus type I-induced disease: pathways to cancer and neurodegeneration.
Retroviral infection is associated with a number of pathologic abnormalities, including a variety of cancers, immunologic diseases, and neurologic disorders. Shortly after its discovery in 1980, human T cell leukemia virus type I (HTLV-I) was found to be the etiologic agent of both adult T cell leukemia (ATL) and HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP), a neurologic disease characterized by demyelinating lesions in both the brain and the spinal cord. Approximately 5-10% of HTLV-I-infected individuals develop either ATL or HAM/TSP. Interestingly, the two diseases have vastly different pathologies and have rarely been found to occur within the same individual. While a number of host and viral factors including virus strain, viral load, and HLA haplotype have been hypothesized to influence disease outcome associated with HTLV-I infection, the relative contributions of such factors to disease pathogenesis have not been fully established. Recent research has suggested that the route of primary viral infection may dictate the course of disease pathogenesis associated with HTLV-I infection. Specifically, mucosal exposure to HTLV-I has been associated with cases of ATL, while primary viral infection based in the peripheral blood has been correlated with progression to HAM/TSP. However, the cellular and molecular mechanisms regulating disease progression resulting from primary viral invasion remain to be elucidated. Although a variety of factors likely influence these mechanisms, the differential immune response mounted by the host against the incoming virus initiated in either the peripheral blood or the mucosal compartments likely plays a key role in determining the outcome of HTLV-I infection. It has been proposed that the route of infection and size of the initial viral inoculum allows HTLV-I to infect different target cell populations, in turn influencing the breadth of the immune response mounted against HTLV-I and affecting disease pathogenesis. A model of HTLV-I-induced disease progression is presented, integrating information regarding the role of several host and viral factors in the genesis of both neoplasia and neurologic disease induced following HTLV-I infection, focusing specifically on differential viral invasion into the bone marrow (BM) and the influence of this event on the virus-specific CD8(+) cytotoxic T lymphocyte (CTL) response that is initiated following HTLV-I infection.


Visual pathways and acuity hooded rats.
Three experiments on the effects of lesions of the visual system on contrast-detection in hooded rats are described, in which the ability of rats to detect stationary high-contrast square-wave gratings of various fundamental frequencies presented in the central visual field was measured before and after operation. The results suggested the following conclusions: (i) The pathways from retina to striate cortex via dorsal lateral geniculate nucleus (dLGN) conveys information about high spatial frequencies sufficient for normal detection of these gratings, that is up to about 1 cycles/deg. It may be the only pathway to carry this information, and may thus play a unique role in the analysis of fine detail. The high-frequency information is probably relayed from striate cortex to extrastriate cortex, rather than to subcortical sites. (ii) The superior colliculus receives information from the retina up to at least 0.7 cycles/deg, which it then relays to extrastriate visual cortex, probably via the lateral posterior nucleus of the thalamus. (iii) Neither the projections from superior colliculus to other, non-thalamic sites nor the remaining pathways from the retina (e.g. to ventral LGN) appear to carry contrast information higher than 0.3 cycles/deg. These sets of projections therefore do not appear to be used for precise analysis of stationary scenes. These findings suggest that there are considerable similarities between the visual systems of rats and other mammals with respect to the routing of information about stationary spatial contrast, and may help to explain the results of some experiments that have used tasks besides contrast-detection to assess the visual capacities of rats after lesions.


[Bladder tumor. Presentation of 2 patients with soft tissue involvement].
To describe two uncommon cases of bladder tumor metastasizing to soft tissues. Two patients with bladder tumor and cystic metastases in the abdominal wall are described. The cysts were easily palpable and were confirmed histologically. Soft tissues, like the abdominal wall, are unusual sites of metastasis of bladder tumor. The foregoing, however, should be considered in order to make an early diagnosis.


[Impacts of illness experiences on middle-aged health care: a cancer example].
Middle-aged couples play multiple family roles. In addition to supporting each other, they also act as caregivers for their children and elderly parents. When they are diagnosed with a severe illness such as cancer, the impacts on the psycho-spiritual aspects of their lives include fear of death and loss of control. While dealing with these and other disease-related issues, they must maintain intimate relationships with their spouses and try to communicate with their children about their health status. In addition to providing individual care, nurses must make the effort to help their patients make adjustments in their family relationships.


The CSN COVID-19 Rapid Response Program.
The coronavirus disease (COVID-19) pandemic has created unprecedented challenges in caring for individuals living with kidney disease. In response to a growing call for up-to-date information and evidence-informed advice, the Canadian Society of Nephrology has established a COVID-19 Rapid Response Team that will leverage existing evidence and national expertise to inform kidney care practices in the COVID-19 era. Given limited published evidence and compressed timelines, formal clinical practice guidelines are not feasible, and we have adopted rapid review methods to instead provide interim guidance across identified priority areas. In this article, we describe the methodological approach that was applied in developing a first iteration of guidance documents addressing clinical and operational aspects of care for patients treated with in-center hemodialysis, home dialysis, those with advanced chronic kidney disease, those with glomerulonephritis, and those with acute kidney injury. We further describe strategies for maintaining ongoing engagement with the renal community to elicit emerging needs and perspectives as the situation unfolds.


Tissue Inhibitor Of Matrix Metalloproteinase-1 Is Required for High-Fat Diet-Induced Glucose Intolerance and Hepatic Steatosis in Mice.
Plasma levels of tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) are elevated in obesity and obesity-related disorders, such as steatosis, but the metabolic role of TIMP-1 is unclear. Here we investigated how the presence or absence of TIMP-1 affected the development of diet-induced glucose intolerance and hepatic steatosis using the Timp1 null mice. Timp1 knockout (TKO) and wild type (TWT) mice were fed chow, high-fat diet (HFD) or intermediate fat and sucrose diet (IFSD). We determined body weight, body composition, lipid content of the liver, energy intake, energy expenditure, oral glucose tolerance, as well as insulin tolerance. In addition, the histology of liver and adipose tissues was examined and expression of selected genes involved in lipid metabolism and inflammation in liver and adipose tissues was determined by RT-qPCR. TKO mice gained less weight and had lower energy efficiency than TWT mice when fed HFD, but not when fed chow or IFSD. Importantly, TKO mice were protected from development of HFD- as well as IFSD-induced glucose intolerance, hepatic steatosis, and altered expression of genes involved in hepatic lipid metabolism and inflammation. Collectively, our results indicate that TIMP-1 contributes to the development of diet-induced hepatic steatosis and glucose intolerance and may be a potential therapeutic target.


Interval and continuous exercise overcome memory deficits related to β-Amyloid accumulation through modulating mitochondrial dynamics.
Exercise is a non-pharmacological strategy that may help to protect against cognitive decline and reduce the risk of Alzheimer's disease. However, the optimal exercise modes for cognitive benefits are controversial. Mitochondrial function has been related to both exercise and cognition. The present study aimed to investigate the effects of two exercise modes on cognitive function and mitochondrial dynamics in APP/PS1 transgenic mice. The results showed that 12-week high-intensity interval training (HIIT) and moderate-intensity continuous training (MICT) could improve exploratory behavior, spatial learning and memory ability of APP/PS1 transgenic mice. Both HIIT and MICT interventions significantly alleviated the hippocampal β-Amyloid (Aβ) burden and mitochondrial fragmentation and improved mitochondrial morphology in hippocampus. Furthermore, both HIIT and MICT interventions down-regulated dynamin-related protein 1 (DRP1) and fission 1 (FIS1), whereas mitofusin 1 (MFN1), mitofusin 2 (MFN2) and optic atrophy 1 (OPA1) were up-regulated. Hippocampal levels of total reactive oxygen species (ROS), malondialdehyde (MDA) and hydrogen peroxide (H2O2) were decreased, whereas activities of superoxide dismutase (SOD) and catalase (CAT) were elevated by HIIT and MICT. The study suggests that both HIIT and MICT alleviate cognitive decline and down-regulat Aβ level in the hippocampus in APP/PS1 transgenic mice, which may be mediated by improvements in mitochondrial morphology and dynamics.


Crystal Structures of TbCatB and rhodesain, potential chemotherapeutic targets and major cysteine proteases of Trypanosoma brucei.
Trypanosoma brucei is the etiological agent of Human African Trypanosomiasis, an endemic parasitic disease of sub-Saharan Africa. TbCatB and rhodesain are the sole Clan CA papain-like cysteine proteases produced by the parasite during infection of the mammalian host and are implicated in the progression of disease. Of considerable interest is the exploration of these two enzymes as targets for cysteine protease inhibitors that are effective against T. brucei. We have determined, by X-ray crystallography, the first reported structure of TbCatB in complex with the cathepsin B selective inhibitor CA074. In addition we report the structure of rhodesain in complex with the vinyl-sulfone K11002. The mature domain of our TbCat*CA074 structure contains unique features for a cathepsin B-like enzyme including an elongated N-terminus extending 16 residues past the predicted maturation cleavage site. N-terminal Edman sequencing reveals an even longer extension than is observed amongst the ordered portions of the crystal structure. The TbCat*CA074 structure confirms that the occluding loop, which is an essential part of the substrate-binding site, creates a larger prime side pocket in the active site cleft than is found in mammalian cathepsin B-small molecule structures. Our data further highlight enhanced flexibility in the occluding loop main chain and structural deviations from mammalian cathepsin B enzymes that may affect activity and inhibitor design. Comparisons with the rhodesain*K11002 structure highlight key differences that may impact the design of cysteine protease inhibitors as anti-trypanosomal drugs.


Quality assured measurements of animal building emissions: odor concentrations.
Standard protocols for sampling and measuring odor emissions from livestock buildings are needed to guide scientists, consultants, regulators, and policy-makers. A federally funded, multistate project has conducted field studies in six states to measure emissions of odor, coarse particulate matter (PM(10)), total suspended particulates, hydrogen sulfide, ammonia, and carbon dioxide from swine and poultry production buildings. The focus of this paper is on the intermittent measurement of odor concentrations at nearly identical pairs of buildings in each state and on protocols to minimize variations in these measurements. Air was collected from pig and poultry barns in small (10 L) Tedlar bags through a gas sampling system located in an instrument trailer housing gas and dust analyzers. The samples were analyzed within 30 hr by a dynamic dilution forced-choice olfactometer (a dilution apparatus). The olfactometers (AC'SCENT International Olfactometer, St. Croix Sensory, Inc.) used by all participating laboratories meet the olfactometry standards (American Society for Testing and Materials and European Committee for Standardization [CEN]) in the United States and Europe. Trained panelists (four to eight) at each laboratory measured odor concentrations (dilution to thresholds [DT]) from the bag samples. Odor emissions were calculated by multiplying odor concentration differences between inlet and outlet air by standardized (20 degrees C and 1 atm) building airflow rates.


Analysis of the metabolic turnover of the individual phosphate groups of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate. Validation of novel analytical techniques by using 32P-labelled lipids from erythrocytes.
We have developed methods that yield estimates of the 32P content of each of the individual phosphate groups of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate, thus extending the information available from studies of the labelling of these lipids in intact cells or membrane preparations. The analyses are undertaken with the deacylated lipids. Assay of the 5-phosphate of phosphatidylinositol 4,5-bisphosphate is achieved by the use, under conditions of first-order kinetics, of a 5-phosphate-specific phosphomonoesterase present in isolated erythrocyte membranes [Downes, Mussat & Michell (1982) Biochem. J. 203, 169-177]. Assay of the 4-phosphate of phosphatidylinositol 4-phosphate and of the total monoester phosphate content (4-phosphate plus 5-phosphate) of phosphatidylinositol 4,5-bisphosphate employs alkaline phosphatase from bovine intestine. The radioactivity of the 1-phosphate is that remaining as organic phosphate after exhaustive alkaline phosphatase treatment. The methodology has been validated by using lipids from human erythrocytes: these contain no 32P in their 1-phosphate. These methods should be of substantial value in studies of the many cells that show rapid hormonal perturbations of phosphatidylinositol 4,5-bisphosphate metabolism.


Resistin stimulates PC-3 prostate cancer cell growth through stimulation of SOCS3 and SOCS5 genes.
Resistin and suppressors of cytokine signaling (SOCSs) have been reported to regulate prostate cancer (PCa) cell proliferation and survival, respectively. Whether any of the SOCS molecules mediate the mitogenic effect of resistin on PCa cells is unknown. Using PC-3 human PCa cells, we found that resistin upregulates the expression of SOCS3 and SOCS5 mRNA, but not SOCS7 mRNA, in a dose- and time-dependent manner. The resistin-induced increases in SOCS3 and SOCS5 expression and cell proliferation were prevented by pretreatment with specific inhibitors of the TLR4, ERK, p38 MAPK, JNK, PI3K, and JAK2 proteins. However, pretreatment with a TLR2 inhibitor had no effect on resistin-mediated SOCS3 and SOCS5 expression. In addition, the effects of resistin on SOCS3, SOCS5, and SOCS7 mRNA levels were cell type-specific. Overexpression of either SOCS3 or SOCS5 enhanced further resistin-stimulated growth of PC-3 cells, whereas silencing SOCS3 or SOCS5 antagonized resistin-increased cell growth. Further PCa tissue analysis demonstrated higher levels of RETN, TLR4, SOCS3, and SOCS5 mRNAs in cancer tissues than benign prostate hyperplasia and indicated positive correlations among RETN, TLR4, and SOCS5. These data suggest that SOCS5, TLR4, and, to a lesser extent, SOCS3 can mediate the mitogenic effect of resistin on PC-3 PCa cells.


Postischemic free radical production in the venous blood of the regionally ischemic swine heart. Effect of deferoxamine.
We tested the hypothesis that secondarily produced free radicals can be detected in venous coronary effluent without the need for direct exposure of postischemic tissue to the spin trapping agent alpha-phenyl-tert-butylnitrone (PBN). The left anterior descending coronary artery (LAD) of pigs was ligated for 15, 30, 40, or 60 minutes, and the tissue was subsequently reperfused for 60 minutes. Venous effluent (6.5 ml) from the risk area was withdrawn sequentially at 1.5-minute intervals during reperfusion. The effluent blood was immediately infused (4.5 ml/min) with an isotonic saline solution containing 120 mM PBN: Preischemic control effluent samples were collected in an identical fashion. Plasma from each sample was extracted in organic solvent and subsequently analyzed by electron spin resonance (ESR) spectroscopy. Another group of pigs received an infusion of the metal chelator deferoxamine mesylate (25 mg/kg/hr) into the right atrium starting 1 hour before the 40-minute ligation and continuing throughout ligation and reperfusion. We were able to demonstrate the postischemic production of ESR signals for PBN adduct(s) from untreated hearts having spectral characteristics similar to an alkoxyl adduct (PBN-RO.; hyperfine splitting constants for beta-hydrogen [alpha H] = 2.0-2.25 G; nitrogen [alpha N] = 13.5-13.75 G). The reperfusion time course of PBN adduct production had a unique pattern: 1) multiple low-level bursts during the initial 15 minutes of reperfusion, and 2) a prominent PBN adduct signal during a relatively late time (20-25 minutes) of reperfusion. Total postischemic PBN adduct production rose with increasing duration (15-60 minutes) of ischemia and was associated with a progressive elevation of total lactate dehydrogenase in the effluent. Infusion of deferoxamine markedly diminished PBN adduct production as well as total release of lactate dehydrogenase. These data suggest the potential feasibility of using an ex vivo ESR spin trapping technique in blood-perfused models of cardiovascular injury and that chelatable free iron contributes to the production of alkoxyl radicals.


Urinary tract infection localization in women.
The site of urinary infection in 51 adult women was determined by the bladder-washout technique (BW). Of the 37 patients with proved upper-tract infection shown by BW, 16 had upper-tract symptoms, three had lower-tract symptoms, and 18 were asymptomatic; 31 had antibody-coated bacteria (ACB), and six had no ACB. Of 14 patients with proved lower-tract infection shown by BW, three had upper-tract symptoms, five had lower-tract symptoms, and six were asymptomatic; none had ACB. All 31 women whose infecting bacteria were coated with antibody proved to have an upper-tract infection shown by BW. A subset of the population of adult women with upper-tract infection have organisms emanating from their ureters at BW localization, which are not coated with antibody. Physicians must be cautious in interpreting results that are negative for ACB.


Mendelian factors underlying quantitative traits in tomato: comparison across species, generations, and environments.
As part of ongoing studies regarding the genetic basis of quantitative variation in phenotype, we have determined the chromosomal locations of quantitative trait loci (QTLs) affecting fruit size, soluble solids concentration, and pH, in a cross between the domestic tomato (Lycopersicon esculentum Mill.) and a closely-related wild species, L. cheesmanii. Using a RFLP map of the tomato genome, we compared the inheritance patterns of polymorphisms in 350 F2 individuals with phenotypes scored in three different ways: (1) from the F2 progeny themselves, grown near Davis, California; (2) from F3 families obtained by selfing each F2 individual, grown near Gilroy, California (F3-CA); and (3) from equivalent F3 families grown near Rehovot, Israel (F3-IS). Maximum likelihood methods were used to estimate the approximate chromosomal locations, phenotypic effects (both additive effects and dominance deviations), and gene action of QTLs underlying phenotypic variation in each of these three environments. A total of 29 putative QTLs were detected in the three environments. These QTLs were distributed over 11 of the 12 chromosomes, accounted for 4.7-42.0% of the phenotypic variance in a trait, and showed different types of gene action. Among these 29 QTLs, 4 were detected in all three environments, 10 in two environments, and 15 in only a single environment. The two California environments were most similar, sharing 11/25 (44%) QTLs, while the Israel environment was quite different, sharing 7/20 (35%) and 5/26 (19%) QTLs with the respective California environments. One major goal of QTL mapping is to predict, with maximum accuracy, which individuals will produce progeny showing particular phenotypes. Traditionally, the phenotype of an individual alone has been used to predict the phenotype of its progeny. Our results suggested that, for a trait with low heritability (soluble solids), the phenotype of F3 progeny could be predicted more accurately from the genotype of the F2 parent at QTLs than from the phenotype of the F2 individual. For a trait with intermediate heritability (fruit pH), QTL genotype and observed phenotype were about equally effective at predicting progeny phenotype. For a trait with high heritability (mass per fruit), knowing the QTL genotype of an individual added little if any predictive value, to simply knowing the phenotype. The QTLs mapped in the L. esculentum X L. cheesmanii F2 appear to be at similar locations to many of those mapped in a previous cross with a different wild tomato (L. chmielewskii).(ABSTRACT TRUNCATED AT 400 WORDS)


The neurobiology of cannabinoid analgesia.
The discovery of cannabinoid receptors and their putative endogenous ligands raises questions as to the nature of the effects produced by cannabinoids on neural circuits that mediate pain and whether endogenous cannabinoids produced by the brain or in the periphery serve naturally to modulate pain. A sizable body of previous work showed that cannabinoid agonists suppress pain behavior in a variety of models of acute and chronic pain. However, at appropriate doses, cannabinoids also profoundly suppress motor behavior (see Sañudo-Peña et al., this volume), which complicates the interpretation of behavioral analgesia since a motor response is the endpoint of virtually all such studies. Studies conducted in this laboratory used biochemical and neurophysiological measures to determine whether cannabinoids suppress nociceptive neurotransmission. The results showed that cannabinoids suppress nociceptive neurotransmission at the level of the spinal cord and the thalamus. These effects are reversible, receptor mediated, selective for painful as opposed to nonpainful somatic stimuli, and track the behavioral analgesia both in time course and potency.