Fluorouracil: biochemistry and pharmacology.
Fluorouracil (5FU) is still considered the most active antineoplastic agent in the treatment of advanced colorectal cancer. The drug needs to be converted to the nucleotide level in order to exert its effect. It can be incorporated into RNA leading to interference with the maturation of nuclear RNA. However, its conversion to 5-fluoro-2'deoxy-5' monophosphate (FdUMP) leading to inhibition of thymidylate synthase (TS) and subsequently of DNA synthesis, is considered to be its main mechanism of action. In the presence of a folate cofactor a covalent ternary complex is formed, the stability of which is the main determinant of the action of 5FU. Resistance against 5FU can be mainly attributed to aberrations in its metabolism or to alterations of TS, eg, gene amplification, altered kinetics in respect to nucleotides or folates. Biochemical modulation of 5FU metabolism can be applied to overcome resistance against 5FU. A variety of normal purines, pyrimidines, and other antimetabolites have been studied in this respect, but only some of them have been clinically successful. Delayed administration of uridine has recently been shown to "rescue" mice and patients from toxicity, while pretreatment with leucovorin is the most promising combination to enhance the therapeutic efficacy. 5FU is frequently administered in an intravenous (IV) injection, and shows a rapid distribution and a triphasic elimination. The nonlinearity of 5FU pharmacokinetics is related to saturation of its degradation. Continuous infusion of 5FU led to different kinetics. Regional administration, such as hepatic artery infusion, offers a way to achieve higher drug concentrations in liver metastases and is accompanied by lower systemic concentration. The current status of the biochemical and pharmacokinetic data is reviewed.


The use of azithromycin and pyrimethamine for treatment of cerebral toxoplasmosis in human immunodeficiency virus-infected patients: a systematic review.
Toxoplasma gondii is a parasite that is widely distributed around the globe and can cause brain inflammation, particularly in immunosuppressed patients such as those diagnosed with human immunodeficiency virus (HIV). This paper reviews the efficacy of azithromycin and pyrimethamine combination therapy for cerebral toxoplasmosis in patients with HIV. The scope of the studies included in this review was limited from 1992 to 2022, with studies primarily being randomized, controlled clinical trials available on online scientific journal databases. The authors screened eligible records for review, removing those that did not fit the inclusion and exclusion criteria. The risk of bias of the extracted data was analyzed through the Cochrane risk-of-bias tool for randomized trials. A broad search of major online databases such as PubMed, Medline, Google Scholar, and Cochrane using keywords, limit fields, and Boolean operators yielded 3,130 articles. After thoroughly screening the search results, two studies were included in this review. Results from the studies included in the review demonstrate that the combination therapy of azithromycin and pyrimethamine is favorable for cerebral toxoplasmosis. However, the net response is less effective than the standard treatment regimen (pyrimethamine and sulfadiazine). The combination therapy of azithromycin and pyrimethamine is less effective than the standard treatment regimen for maintenance therapy for cerebral toxoplasmosis; thus, administering these medications for this indication must be met with caution.


Combined epidemiological and genomic analysis of nosocomial SARS-CoV-2 infection early in the pandemic and the role of unidentified cases in transmission.
To analyse nosocomial transmission in the early stages of the coronavirus 2019 (COVID-19) pandemic at a large multisite healthcare institution. Nosocomial incidence is linked with infection control interventions. Viral genome sequence and epidemiological data were analysed for 574 consecutive patients, including 86 nosocomial cases, with a positive PCR test for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during the first 19 days of the pandemic. Forty-four putative transmission clusters were found through epidemiological analysis; these included 234 cases and all 86 nosocomial cases. SARS-CoV-2 genome sequences were obtained from 168/234 (72%) of these cases in epidemiological clusters, including 77/86 nosocomial cases (90%). Only 75/168 (45%) of epidemiologically linked, sequenced cases were not refuted by applying genomic data, creating 14 final clusters accounting for 59/77 sequenced nosocomial cases (77%). Viral haplotypes from these clusters were enriched 1-14x (median 4x) compared to the community. Three factors implicated unidentified cases in transmission: (a) community-onset or indeterminate cases were absent in 7/14 clusters (50%), (b) four clusters (29%) had additional evidence of cryptic transmission, and (c) in three clusters (21%) diagnosis of the earliest case was delayed, which may have facilitated transmission. Nosocomial cases decreased to low levels (0-2 per day) despite continuing high numbers of admissions of community-onset SARS-CoV-2 cases (40-50 per day) and before the impact of introducing universal face masks and banning hospital visitors. Genomics was necessary to accurately resolve transmission clusters. Our data support unidentified cases-such as healthcare workers or asymptomatic patients-as important vectors of transmission. Evidence is needed to ascertain whether routine screening increases case ascertainment and limits nosocomial transmission.


A binary-BAC system for plant transformation with high-molecular-weight DNA.
A binary-BAC (BIBAC) vector suitable for Agrobacterium-mediated plant transformation with high-molecular-weight DNA was constructed. A BIBAC vector is based on the bacterial artificial chromosome (BAC) library vector and is also a binary vector for Agrobacterium-mediated plant transformation. The BIBAC vector has the minimal origin region of the Escherichia coli F plasmid and the minimal origin of replication of the Agrobacterium rhizogenes Ri plasmid, and thus replicates as a single-copy plasmid in both E. coli and in A. tumefaciens. The T-DNA of the BIBAC vector can be transferred into the plant nuclear genome. As examples, a 30-kb yeast genomic DNA fragment and a 150-kb human genomic DNA fragment were inserted into the BIBAC vector; these constructs were maintained in both E. coli and A. tumefaciens. In order to increase the efficiency of transfer of unusually large BIBAC T-DNAs, helper plasmids that carry additional copies of A. tumefaciens virulence genes virG and virE were constructed. These helper plasmids are compatible with, and can be present in addition to, the BIBAC vector in the A. tumefaciens host. This report details the components of the BIBAC system, providing information essential to the general understanding and the application of this new technology.


Academic etiquette for the nephrologist.
Academic etiquette is an underexplored and underemphasized topic. Yet, a collegial atmosphere is essential to accomplish the missions of an academic medical center. Appropriate social, personal, and cultural behaviors are not only desirable, but they are also necessary to practice and emulate. As faculty in an academic center, one may want to share these thoughts with peers, students, and other healthcare providers. As a trainee, one may want dedicated lectures on this topic and to observe the behaviors modeled by mentors. This article attempts to outline principles of appropriate etiquette in the academic workplace. These include issues of loyalty, collegiality, and collaboration in daily activities from research to clinical care. Approaching personal interactions with grace and integrity can be an essential tool in the pursuit of academic excellence and success.


What Turkish nurses know and do about skin cancer and sun protective behavior.
Skin cancer is a global health problem that can be prevented by protective behavior promoted by nurses. In Turkey, only few studies have examined current knowledge of nurses related to skin cancer and to reveal their attitudes towards sun exposure and current protective behavior. This descriptive cross-sectional study was performed from March 11 to May 30, 2013 with a study sample of 310 nurses working in two state hospitals located in Karabuk and Safranbolu. Mean age of the nurses was 30.3±6.89 years and 96.1% of them had graduated with a Bachelor degree. The participants were predominantly female (n=284). Knowledge level about skin cancer was significantly higher for females compared to males. Nurses do not have sufficient knowledge about skin cancer and are not adequately protecting themselves from ultraviolet light, the primary risk factor for skin cancer. While they demonstrated a responsible attitude towards avoiding sunburn and the need for adequate sun protection, they do not fully appreciate the extent to which the sun can cause skin cancer and that they lack full understanding about the need to protect the skin from burning and to avoid long term sun exposure in Turkey.


CD11c+ alveolar macrophages are a source of IL-23 during lipopolysaccharide-induced acute lung injury.
Acute lung injury (ALI) is a severe pulmonary disease causing high numbers of fatalities worldwide. Innate immune responses are an integral part of the pathophysiologic events during ALI. Interleukin 23 (IL-23) is a proinflammatory mediator known to direct the inflammatory responses in various settings of infection, autoimmunity, and cancer. Interleukin 23 has been associated with proliferation and effector functions in T(H)17 cells. Surprisingly, little is known about production of IL-23 during ALI. In this study, we found expression of mRNA for IL-23p19 to be 10-fold elevated in lung homogenates of C57BL/6 mice after lipopolysaccharide (LPS)-induced ALI. Likewise, concentrations of IL-23 significantly increased in bronchoalveolar lavage fluids. Experiments with IL-23-deficient mice showed that endogenous IL-23 was required for production of IL-17A during LPS-ALI. CD11c-diphtheria toxin receptor transgenic mice were used to selectively deplete CD11c cells, the data suggesting that IL-23 production is dependent at least in part on CD11c cells during ALI. No alterations of IL-23 levels were observed in Rag-1-deficient mice as compared with wild-type C57BL/6 mice following ALI. The mouse alveolar macrophage cell line, MH-S, as well as primary alveolar macrophages displayed abundant surface expression of CD11c. Activation of these macrophages by LPS resulted in release of IL-23 in vitro. Our findings identify CD11c macrophages in the lung are likely an important source of IL-23 during ALI, which may be helpful for better understanding of this disease.


T30177, an oligonucleotide stabilized by an intramolecular guanosine octet, is a potent inhibitor of laboratory strains and clinical isolates of human immunodeficiency virus type 1.
T30177, an oligonucleotide composed of only deoxyguanosine and thymidine, is 17 nucleotides in length and contains single phosphorothioate internucleoside linkages at its 5' and 3' ends for stability. This oligonucleotide does not share significant primary sequence homology with or possess any complementary (antisense) sequence motifs to the human immunodeficiency virus type 1 (HIV-1) genome. T30177 inhibited replication of multiple laboratory strains of HIV-1 in human T-cell lines, peripheral blood lymphocytes, and macrophages. T30177 was also found to be capable of inhibiting multiple clinical isolates of HIV-1 and preventing the cytopathic effect of HIV-1 in primary CD4+ T lymphocytes. In assays with human peripheral blood lymphocytes there was no observable toxicity associated with T30177 at the highest concentration tested (100 microM), while the median inhibitory concentration was determined to be in the range of 0.1 to 1.0 microM for the clinical isolates tested, resulting in a high therapeutic index for this drug. In temporal studies, the kinetics of addition of T30177 to infected cell cultures indicated that, like the known viral adsorption blocking agents dextran sulfate and Chicago sky blue, T30177 needed to be added to cells during or very soon after viral infection. However, analysis of nucleic acids extracted at 12 h postinfection from cells treated with T30177 at the time of virus infection established the presence of unintegrated viral cDNA, including circular proviral DNA, in the treated cells. In vitro analysis of viral enzymes revealed that T30177 was a potent inhibitor of HIV-1 integrase, reducing enzymatic activity by 50% at concentrations in the range of 0.050 to 0.09 microM. T30177 was also able to inhibit viral reverse transcriptase activity; however, the 50% inhibitory value obtained was in the range of 1 to 10 microM, depending on the template used in the enzymatic assay. No observable inhibition of viral protease was detected at the highest concentration of T30177 used (10 microM). In experiments in which T30177 was removed from infected cell cultures at 4 days post-HIV-1 infection, total suppression of virus production was observed for more than 27 days. PCR analysis of DNA extracted from cells treated in this fashion was unable to detect the presence of viral DNA 11 days after removal of the drug from the infected cell cultures. The ability of T30177 to inhibit both laboratory and clinical isolates of HIV-1 and the experimental data which suggest that T30177 represents a novel class of integrase inhibitors indicate that this compound is a viable candidate for evaluation as a therapeutic agent against HIV-1 in humans.


Internal jugular vein valve incompetence and intracranial venous anatomy in transient global amnesia.
Recently a causal relation between internal jugular vein valve incompetence (IJVVI) and transient global amnesia (TGA) has been suggested. IJVVI is postulated to provoke a transient mesiotemporal ischaemia by venous congestion. This mechanism requires a patent venous pathway from the affected IJV through the transverse sinus, confluens, straight sinus (SS), vein of Galen into the basal vein of Rosenthal and the internal cerebral veins. To study IJVVI in TGA patients in relation to the intracranial venous anatomy. IJVVI was defined if a repeated Valsalva manoeuvre (VM) led to a retrograde jugular flow detected by extracranial duplex ultrasound. Non-contrast venous MR angiography (MRA) was performed to analyse intracranial drainage patterns of the SS in relation to the side of the IJVVI. SS drainage was differentiated into three groups: predominantly right, left, and bilateral drainage. Ultrasound studies were performed in 25 TGA patients and 85 age matched controls. Twenty patients underwent venous MRA. Sixty eight per cent of patients and 33% of controls showed unilateral or bilateral IJVVI (p = 0.0025). In 36% of patients a TGA preceding VM was reported. Drainage pattern of SS and side of IJVVI corresponded in five of eight patients (63%) with VM and four of 12 patients without VM (33%, p = 0.0994). Our study confirms the significantly higher prevalence of IJVVI in TGA patients. However, no specific IJVVI related intracranial venous drainage patterns could be found to further support the hypothesis of a direct causal relation between IJVVI and TGA.


The activin receptor-like kinase 6 Booroola mutation enhances suppressive effects of bone morphogenetic protein 2 (BMP2), BMP4, BMP6 and growth and differentiation factor-9 on FSH release from ovine primary pituitary cell cultures.
Bone morphogenetic proteins (BMPs) have been shown to influence the regulation of FSH synthesis and secretion at the level of the pituitary. Primary pituitary cells were harvested and cultured from Booroola ewes homozygous for a mutation in activin receptor-like kinase 6 (ALK6) also known as BMP receptor IB (BMPRIB), and from wild-type (WT) ewes to determine if the mutation caused alterations in FSH secretion in vitro. The cells were collected 24 h following induction of luteolysis and cultured for 72 h prior to being challenged for 24 h with BMP2, BMP4, BMP6, growth and differentiation factor-9 (GDF9), transforming growth factor-beta 1, activin-A and GnRH. The levels of FSH and LH were measured by RIA and then compared with the untreated controls. Primary pituitary cell cultures from Booroola ewes secreted less FSH than WT cells in the presence of BMP2, BMP4 and BMP6. These BMPs did not affect the FSH stores within the cells, or the levels of LH released. GDF9 appeared to act in a BMP-like manner by suppressing FSH secretion. The ALK6 receptor however, was not found to co-localise with gonadotroph cells in either Booroola or WT pituitary tissues. These findings imply that the increased sensitivity of Booroola cells to BMP2, BMP4, BMP6 and GDF9 cannot be due to the direct action of the ALK6 mutant Booroola receptor in the cells that synthesise FSH.


Reliability and validity of the Dutch version of the inventory of traumatic grief (ITG).
The psychometric properties of the Dutch version of the Inventory of Traumatic Grief (ITG) were investigated in two studies with bereaved adults who had suffered the loss of a first-degree relative. In Study 1, exploratory factor analysis indicated that the items of the ITG clustered together into one underlying factor. In addition, the internal consistency of the ITG and its short-term temporal stability were found to be high. In Study 2 it was found that the ITG exhibited adequate discriminative, concurrent, and construct validity. Furthermore, an ITG cutoff score for a diagnosis of traumatic grief was determined, with a sensitivity of 86% and a specificity of 76%, providing evidence in favor of the predictive validity of the ITG.


Mifepristone Increases Thyroid Hormone Requirements in Patients With Central Hypothyroidism: A Multicenter Study.
Mifepristone is a glucocorticoid and progesterone receptor blocker that can be used for patients with hyperglycemia and Cushing syndrome in whom surgery failed to achieve remission or who were ineligible for surgery. We report a case series of patients with Cushing disease (CD) and central hypothyroidism that presented with increased levothyroxine requirements during mifepristone therapy. Retrospective longitudinal case series of patients with CD and central hypothyroidism treated with mifepristone in a retrospective database at four pituitary centers in the United States. Five patients with CD were found, all women, median age 50 (interquartile range 47 to 64.5). They received mifepristone because no adequate response or intolerance to other drugs was observed. Mifepristone initiation was associated with a decrease in free thyroxine levels, mandating a dose increase of a median 1.83 (1.71 to 3.5) times the initial dose of levothyroxine to achieve normal levels. Weight loss was seen in four of five patients, ranging from 3.2 to 42.6 kg in up to 54 months of follow-up. Although the mechanism behind the decrease in thyroid hormone level is unknown, intestinal malabsorption, decreased residual thyroid function and increased inactivation of T4 via deiodinases are all potential causes. Whereas therapies for hypercortisolism aim to decrease features of hypercortisolemia such as weight gain and depression, hypothyroidism can hamper these goals. This case series raises awareness on the importance of assessment of thyroid status in patients receiving mifepristone to optimize clinical outcomes.


Natural killer cell immunotherapy to target stem-like tumor cells.
Advances in cancer immunotherapy are leading to its increasing and successful application for the treatment of solid-tissue cancers. Despite the recent advances there are still significant barriers, in particular, evidence for significant tumor heterogeneity, both genetic and epigenetic that limit long-term efficacy. Subpopulations of "stem-like" tumor cells have been identified in nearly all human malignancies based on both morphologic and functional criteria. Also called cancer stem cells or CSCs, these quiescent cells display enhanced tumorigenic potential and are capable of repopulating tumors in the wake of traditional cytoreductive therapies. These CSCs may be best targeted via immunotherapy. Our lab has identified activated natural killer (NK) cell-based therapy as an effective method to target CSCs particularly after radiation therapy for solid tumors. Using a variety of in vitro and in vivo methods, including the utilization of primary tumor tissue and patient-derived xenografts, we observed that autologous and allogeneic NK cells possess the ability to preferentially kill stem-like cells or CSCs from freshly isolated patient samples representing a broad spectrum of tumor types, including pancreatic cancers, breast cancers, and sarcomas. The results indicated that CSCs express stress ligand molecules capable of being targeted by NKG2D on NK cells and that prior radiation therapy can both deplete the cycling non-CSCs bulk tumor population and upregulate these stress ligands on the CSC making this an effective combination approach.


Evaluation of a DNA Aβ42 vaccine in adult rhesus monkeys (Macaca mulatta): antibody kinetics and immune profile after intradermal immunization with full-length DNA Aβ42 trimer.
Aggregated amyloid-β peptide 1-42 (Aβ42), derived from the cellular amyloid precursor protein, is one of the pathological hallmarks of Alzheimer's disease (AD). Although active immunization against Aβ42 peptide was successful in AD mouse models and led to removal of plaques and improved memory, a similar clinical trial in humans (Aβ42 peptide immunization with QS-21 adjuvant) was stopped in phase II, when 6% of the treated patients developed encephalitis. Currently ongoing passive immunizations with the injection of preformed monoclonal antibodies against different epitopes within the Aβ1-42 peptide, which do not lead to activation of the immune system, have shown some effects in slowing AD pathology. Active DNA Aβ42 immunizations administered with the gene gun into the skin are noninflammatory because they activate a different T-cell population (Th2) with different cytokine responses eliciting a different humoral immune response. We present our findings in rhesus macaques that underwent the DNA Aβ42 immunization via gene gun delivery into the skin. Six rhesus monkeys received two different doses of a DNA Aβ42 trimer vaccine. The humoral immune response was analyzed from blood throughout the study, and cellular immune responses were determined in peripheral blood mononuclear cells (PBMCs) after three and six immunizations. DNA Aβ42 trimer immunization led to high titer antibody responses in the nonhuman primate (NHP) model. Antibodies generated in the rhesus monkeys following DNA Aβ42 immunization detected amyloid plaques consisting of human Aβ42 peptide in the brain of the triple-transgenic AD mouse model. T-cell responses showed no interferon (IFN)-γ- and interleukin (IL)-17-producing cells from PBMCs in Enzyme-Linked ImmunoSpot assays after three immunization time points. At six immunization time points, IFN-γ- and IL-17-producing cells were found in immunized animals as well as in control animals and were thus considered nonspecific and not due to the immunization regimen. IFN-γ and IL-17 secretion in response to Aβ42 peptide restimulation became undetectable after a 3-month rest period. Intradermal DNA Aβ42 immunization delivered with the gene gun produces a high antibody response in NHPs and is highly likely to be effective and safe in a clinical AD prevention trial in patients.


Topographic forcing of the atmosphere and a rapid change in the length of day.
During June to September 1992, a special campaign was held to measure rapid changes in Earth's rotation rate and to relate these measurements to variations in the atmosphere's angular momentum, due principally to changes in zonal winds. A strong rise in both length of day and atmospheric momentum during a particular 6-day subperiod is documented, and this example of a short-period perturbation is identified with a specific regional coupling mechanism. Mountain torques within the southern tropics appear to account for most of the rapid momentum transfer between the solid Earth and atmosphere, with those across South America especially important.


Relationship between pretransplant noncompliance and posttransplant outcomes in renal transplant recipients.
Approximately 5% to 18% of kidney transplant recipients do not comply with their posttransplant medical treatment. This study examined the relationship between pretransplant noncompliance and posttransplant outcomes. Using a longitudinal retrospective chart audit, pretransplant and posttransplant data were collected for 126 kidney transplant recipients over a 3-year period. Sixty-one percent of those identified as noncompliant before transplant lost their graft or died after transplant. Significant relationships between pretransplant noncompliance and graft loss and between pre- and posttransplant noncompliance were found. Clinicians must identify those with pretransplant noncompliance, as they are at risk for poor outcomes and might benefit from an intensive posttransplant follow-up regimen.


Surgery for levodopa-induced dyskinesias.
The effects of surgical interventions for levodopa-induced dyskinesias (LIDs) in Parkinson's disease are reviewed. Since the introduction of levodopa, thalamotomy has been reported to have variable effects on LIDs. Striking benefit has been demonstrated, and efficacy is probably dependent on the size and location of the lesion. However, it remains unclear whether it is an effective treatment for all types of LIDs. Currently, pallidotomy is probably the most reliable and effective surgical treatment for LIDs, and will continue to play an important role in its management until other treatments become more widely available. Deep brain stimulation is an extremely exciting mode of therapy, particularly in the subthalamic nucleus and the globus pallidus, and early results of its use in the treatment of LIDs are promising. The effects of cerebral transplantation, still an experimental technique, on LIDs are inconsistent and controversial, and there is little reliable evidence that gamma knife radiosurgery can be safely applied to parkinsonian patients for the treatment of LIDs.


Three-dimensional structure of the adenovirus major coat protein hexon.
The three-dimensional crystal structure of the adenovirus major coat protein is presented. Adenovirus type 2 hexon, at 967 residues, is now the longest polypeptide whose structure has been determined crystallographically. Taken with our model for hexon packing, which positions the 240 trimeric hexons in the capsid, the structure defines 60% of the protein within the 150 X 10(6) dalton virion. The assembly provides the first details of a DNA-containing animal virus that is 20 times larger than the spherical RNA viruses previously described. Unexpectedly, the hexon subunit contains two similar beta-barrels whose topology is identical to those of the spherical RNA viruses, but whose architectural role in adenovirus is very different. The hexon structure reveals several distinctive features related to its function as a stable protective coat, and shows that the type-specific immunological determinants are restricted to the virion surface.


Utility of functioning in predicting costs of care for patients with mood and anxiety disorders: a prospective cohort study.
Development of payment systems for mental health services has been hindered by limited evidence for the utility of diagnosis or symptoms in predicting costs of care. We investigated the utility of functioning information in predicting costs for patients with mood and anxiety disorders. This was a prospective cohort study involving 102 adult patients attending a tertiary referral specialist clinic for mood and anxiety disorders. The main outcome was total costs, calculated by applying unit costs to healthcare use data. After adjusting for covariates, a significant total costs association was yielded for functioning (e=1.02; 95% confidence interval: 1.01-1.03), but not depressive symptom severity or anxiety symptom severity. When we accounted for the correlations between the main independent variables by constructing an abridged functioning metric, a significant total costs association was again yielded for functioning (e=1.04; 95% confidence interval: 1.01-1.09), but not symptom severity. The utility of functioning in predicting costs for patients with mood and anxiety disorders was supported. Functioning information could be useful within mental health payment systems.


A silicone-based stretchable micropost array membrane for monitoring live-cell subcellular cytoskeletal response.
External forces are increasingly recognized as major regulators of cellular structure and function, yet the underlying mechanism by which cells sense forces and transduce them into intracellular biochemical signals and behavioral responses ('mechanotransduction') is largely undetermined. To aid in the mechanistic study of mechanotransduction, herein we devised a cell stretching device that allowed for quantitative control and real-time measurement of mechanical stimuli and cellular biomechanical responses. Our strategy involved a microfabricated array of silicone elastomeric microposts integrated onto a stretchable elastomeric membrane. Using a computer-controlled vacuum, this micropost array membrane (mPAM) was activated to apply equibiaxial cell stretching forces to adherent cells attached to the microposts. Using the mPAM, we studied the live-cell subcellular dynamic responses of contractile forces in vascular smooth muscle cells (VSMCs) to a sustained static equibiaxial cell stretch. Our data showed that in response to a sustained cell stretch, VSMCs regulated their cytoskeletal (CSK) contractility in a biphasic manner: they first acutely enhanced their contraction to resist rapid cell deformation ('stiffening') before they allowed slow adaptive inelastic CSK reorganization to release their contractility ('softening'). The contractile response across entire single VSMCs was spatially inhomogeneous and force-dependent. Our mPAM device and live-cell subcellular contractile measurements will help elucidate the mechanotransductive system in VSMCs and thus contribute to our understanding of pressure-induced vascular disease processes.


Evaluation of flowable resin composite surfaces eroded by acidic and alcoholic drinks.
The purpose of this study was to evaluate the morphological changes of the surfaces of flowable resins eroded by orange juice and alcohol drinks. The tested products were Beautifil Flow BF02 and BF10, Clearfil Majesty LV, Filtek Supreme XT Flowable Restorative, Unifil LoFlo Plus and Filtek Supreme. Filler percentages of flowable resins were calculated after the latter were incinerated at 750 degrees C. Specimens were shaped into a disk form with a diameter of 10 mm and a thickness of 1 mm. Morphological changes were evaluated for the following types of flowable resin surfaces: polished surface, surfaces eroded by 100% orange juice, wine and whisky. Filler percentages of the tested flowable resins ranged between 42 and 78%. Surface degradation was observed for the specimens immersed in acidic and alcoholic drinks, and it was thought that the lower the filler percentage, the greater was the surface degradation. Decomposition of the matrix resin and fallout of the fillers were observed in flowable resins that eroded with acidic and alcoholic drinks.


Multiple testing to establish superiority/equivalence of a new treatment compared with kappa standard treatments.
In this paper we develop multiple hypotheses testing procedures to compare a new treatment with a set of standard treatments in a clinical trial. The aim is to classify the new treatment with respect to each of the standards, by specifying those to which the new treatment is superior, those to which the new treatment is equivalent and those to which one can establish neither superiority nor equivalence. We propose several stepwise procedures and compare them with respect to their familywise error rates and power. The step-down methods SD1 and SD2 test for superiority first, followed by tests for equivalence for those comparisons where we cannot establish superiority. The step-up methods SU1 and SU2 test for equivalence first, followed by tests for superiority for those comparisons where we can establish at least equivalence. The methods SD3 and SU3 apply the tests for superiority and equivalence in pairs. All the methods require that we specify a threshold value delta > 0 in advance for defining equivalence. In applications where it is not possible to specify a value delta, we can use the method SD1 by testing for superiority first, followed by one-sided confidence limits on the efficacy differences for those comparisons where we cannot establish superiority.


Cognitive control of movement via the cerebellar-recipient thalamus.
The cognitive control of behavior was long considered to be centralized in cerebral cortex. More recently, subcortical structures such as cerebellum and basal ganglia have been implicated in cognitive functions as well. The fact that subcortico-cortical circuits for the control of movement involve the thalamus prompts the notion that activity in movement-related thalamus may also reflect elements of cognitive behavior. Yet this hypothesis has rarely been investigated. Using the pathways linking cerebellum to cerebral cortex via the thalamus as a template, we review evidence that the motor thalamus, together with movement-related central thalamus have the requisite connectivity and activity to mediate cognitive aspects of movement control.


Insights into Modeling Approaches in Chemistry: Assessing Ligand-Protein Binding Thermodynamics Based on Rigid-Flexible Model Molecules.
In the field of chemistry, model compounds find extensive use for investigating complex objects. One prime example of such object is the protein-ligand supramolecular interaction. Prediction the enthalpic and entropic contribution to the free energy associated with this process, as well as the structural and dynamic characteristics of protein-ligand complexes poses considerable challenges. This review exemplifies modeling approaches used to study protein-ligand binding (PLB) thermodynamics by employing pairs of conformationally constrained/flexible model molecules. Strategically designing the model molecules can reduce the number of variables that influence thermodynamic parameters. This enables scientists to gain deeper insights into the enthalpy and entropy of PLB, which is relevant for medicinal chemistry and drug design. The model studies reviewed here demonstrate that rigidifying ligands may induce compensating changes in the enthalpy and entropy of binding. Some "rules of thumb" have started to emerge on how to minimize entropy-enthalpy compensation and design efficient rigidified or flexible ligands.


Chromatin state transitions in the Drosophila intestinal lineage identify principles of cell-type specification.
Tissue homeostasis relies on rewiring of stem cell transcriptional programs into those of differentiated cells. Here, we investigate changes in chromatin occurring in a bipotent adult stem cells. Combining mapping of chromatin-associated factors with statistical modeling, we identify genome-wide transitions during differentiation in the adult Drosophila intestinal stem cell (ISC) lineage. Active, stem-cell-enriched genes transition to a repressive heterochromatin protein-1-enriched state more prominently in enteroendocrine cells (EEs) than in enterocytes (ECs), in which the histone H1-enriched Black state is preeminent. In contrast, terminal differentiation genes associated with metabolic functions follow a common path from a repressive, primed, histone H1-enriched Black state in ISCs to active chromatin states in EE and EC cells. Furthermore, we find that lineage priming has an important function in adult ISCs, and we identify histone H1 as a mediator of this process. These data define underlying principles of chromatin changes during adult multipotent stem cell differentiation.


The antimycobacterial components of hops (Humulus lupulus) and their dereplication.
Bioassay-guided fractionation of a hexane extract of strobile hops (Humulus lupulus) was undertaken to isolate and characterize the antimycobacterial constituents using the fast-growing mycobacterial species Mycobacterium fortuitum. Activity was associated with a low polarity fraction and 1H NMR spectra indicated the presence of a fatty acid mixture with unsaturated components. GC-MS of the derivatives indicated the presence of palmitic, stearic and oleic acids with small quantities of lignoceric, arachidic, behenic and linoleic acids. These compounds were assessed against M. fortuitum and all saturated fatty acids were inactive at concentrations greater than 256 microg/ml, whereas the unsaturated fats oleic and linoleic acids displayed minimum inhibitory concentrations of between 4 and 16 microg/ml against the fast-growing species tested. The widespread occurrence of these components could render screening for antimycobacterials from natural sources highly problematic without adequate dereplication. We propose that GC-MS of derivatised components of lipophilic extracts be a first step before any antimycobacterial bioassay-guided study, as this technique is the method of choice for dereplication of fatty acids.


Secretion and translocation signals and DspB/F-binding domains in the type III effector DspA/E of Erwinia amylovora.
DspA/E is a type III effector of Erwinia amylovora, the bacterial pathogen that causes fire blight disease in roseaceous plants. This effector is indispensable for disease development, and it is translocated into plant cells. A DspA/E-specific chaperone, DspB/F, is necessary for DspA/E secretion and possibly for its translocation. In this work, DspB/F-binding sites and secretion and translocation signals in the DspA/E protein were determined. Based on yeast two-hybrid assays, DspB/F was found to bind DspA/E within the first 210 amino acids of the protein. Surprisingly, both DspB/F and OrfA, the putative chaperone of Eop1, also interacted with the C-terminal 1059 amino acids of DspA/E; this suggests another chaperone-binding site. Secretion and translocation assays using serial N-terminal lengths of DspA/E fused with the active form of AvrRpt2 revealed that at least the first 109 amino acids, including the first N-terminal chaperone-binding motif and DspB/F, were required for efficient translocation of DspA/E, although the first 35 amino acids were sufficient for its secretion and the presence of DspB/F was not required. These results indicate that secretion and translocation signals are present in the N terminus of DspA/E, and that at least one DspB/F-binding motif is required for efficient translocation into plant cells.


Sexual health education in U.S. physician assistant programs.
Since the 1950s, sexual health education in medical schools has been evaluated and reported upon, but there has never been an assessment published about sexual health curricula in U.S. physician assistant (PA) programs. The aim of this study was to gain better understanding of how PA programs cover sexual health topics. Between January and March 2014, 181 accredited PA programs received a mailed survey inquiring about their sexual health curriculum. The survey assessed general sexual health topics; lesbian, gay, bisexual, transgender (LGBT) topics; teaching methods; and the amount of time spent on sexual health education. A total of 106 programs responded (59%). Ten programs offered a required, discrete course on human sexuality. The majority incorporated training into other coursework, which is consistent with most medical schools. LGBT topics were covered less thoroughly than the general sexual health topics. Total amount of time spent on sexual health topics varied widely among programs, from a minimum of 2-4 hours to a maximum of 60 hours, with a median of 12 hours. PA programs in the United States appear to compare favorably with the training offered to medical students in regard to time spent on sexual health education. Transgender issues were least well-covered of all the topics queried.


[Legionnaire's disease contracted in Brussels. One case (author's transl)].
A 71-year-old patient living in a street where major road works were taking place developed diffuse bronchopneumonia with changes in hepatic and renal biochemical tests. Since neutrophilia was moderate and all bacteriological investigations were negative, the patient was given erythromycin and was tested for antibodies directed against Legionella pneumophila. The titers of these antibodies rose from 32 to 2048 after one month. The patient was cured.


[Glutaric aciduria type 1: phenotypic variability. Report of 6 patients].
We report six patients with glutaric aciduria type 1 in four families. The patients had marked clinical variability, even within families. Three of the patients studied were normal until the onset of neurologic abnormalities, that presented as an encephalitis-like illness in the first year of age. One patient had an early and important developmental delay, but never suffered an encephalopathic crisis. Two patients have intellectual preservation; one of them has a mild tremor and choreoathetosis since the first year of age, and the other had only two afebrile seizures in infancy and no other neurologic signs. Three patients are severely handicapped, with a severe dystonic-dyskinetic disorder and unable to even sit. All the six patients have macrocephaly and in all the computed tomography showed enlarged CSF spaces and sulcal separation over the frontal and temporal lobes. Urine organic acids study of all patients showed large quantities of glutaric acid.


Structural Elements of DNA and RNA Eukaryotic Expression Vectors for In Vitro and In Vivo Genome Editor Delivery.
Gene editing with programmable nucleases opens new perspectives in important practice areas, such as healthcare and agriculture. The most challenging problem for the safe and effective therapeutic use of gene editing technologies is the proper delivery and expression of gene editors in cells and tissues of different organisms. Virus-based and nonviral systems can be used for the successful delivery of gene editors. Here we have reviewed structural elements of nonviral DNA- and RNA-based expression vectors for gene editing and delivery methods in vitro and in vivo.


Molecular cloning and analysis of functional cDNA and genomic clones encoding bovine cellular retinoic acid-binding protein.
A recombinant cDNA clone, pCRABP-HS1, encoding cellular retinoic acid-binding protein was isolated from a bovine adrenal cDNA library. COS-7 cells transfected with pCRABP-HS1 produced a biologically active retinoic acid-binding protein molecule of the expected molecular mass (15.5 kDa). RNA blot hybridization analysis using pCRABP-HS1 as a probe revealed a single 1050-nucleotide mRNA species in bovine adrenal, uterus, and testis, tissues that contain the highest levels of retinoic acid-binding activity. No hybridization was detected in RNA extracted from ovary, spleen, kidney, or liver, which contain relatively low levels of cellular retinoic acid-binding protein activity. Analysis of genomic clones isolated from an EcoRI bovine genomic library demonstrated that the bovine cellular retinoic acid-binding protein gene is composed of four exons and three introns. Two putative promoter sequences were identified in the cloned 5' sequence of the gene.


Employee choice of consumer-driven health insurance in a multiplan, multiproduct setting.
To determine who chooses a Consumer-Driven Health Plan (CDHP) in a multiplan, multiproduct setting, and, specifically, whether the CDHP attracts the sicker employees in a company's risk pool. We estimated a health plan choice equation for employees of the University of Minnesota, who had a choice in 2002 of a CDHP and three other health plans--a traditional health maintenance organization (HMO), a preferred provider organization (PPO), and a tiered network product based on care systems. Data from an employee survey were matched to information from the university's payroll system. Chronic illness of the employee or family members had no effect on choice of the CDHP, but such employees tended to choose the PPO. The employee's age was not related to CDHP choice. Higher-income employees chose the CDHP, as well as those who preferred health plans with a national provider panel that includes their physician in the panel. Employees tended to choose plans with lower out-of-pocket premiums, and surprisingly, employees with a chronic health condition themselves or in their family were more price-sensitive. This study provides the first evidence on who chooses a CDHP in a multiplan, multiproduct setting. The CDHP was not chosen disproportionately by the young and healthy, but it did attract the wealthy and those who found the availability of providers more appealing. Low out-of-pocket premiums are important features of health plans and in this setting, low premiums appeal to those who are less healthy.


Src homology region 2-containing protein tyrosine phosphatase-2 (SHP-2) can play a direct role in the inhibitory function of killer cell Ig-like receptors in human NK cells.
The inhibitory forms of killer cell Ig-like receptors (KIR) are MHC class I-binding receptors that are expressed by human NK cells and prevent their attack of normal cells. Substantial evidence indicates that the mechanism of KIR-mediated inhibition involves recruitment of the protein tyrosine phosphatase, Src homology region 2-containing protein tyrosine phosphatase (SHP)-1, to phosphorylated immunoreceptor tyrosine-based inhibitory motifs (ITIMs). However, the functional significance of parallel recruitment of a SHP-1-related phosphatase, SHP-2, to KIR ITIMs has not been addressed. In the present study, our results with mutant forms of a classical KIR, KIR3DL1, show a direct correlation between SHP-2 recruitment and functional inhibition of target cell conjugation and cytotoxicity. In addition, KIR3DL1 inhibition of target cell cytotoxicity is blocked by overexpression of a dominant-negative form of SHP-2. Finally, KIR3DL1 fused directly with the catalytic domain of SHP-2 inhibits both target cell conjugation and cytotoxicity responses. These results strongly indicate that SHP-2 catalytic activity plays a direct role in inhibitory KIR functions, and SHP-2 inhibits NK cell activation in concert with SHP-1.


Does Whole-Body Vibration Training Have a Concurrent Effect on Bone and Muscle Health? A Systematic Review and Meta-Analysis.
Aging results in musculoskeletal disorders, which are a leading cause of disability worldwide. While conventional nonpharmacological treatments have included interventions such as resistance exercise, there are subgroups of people who may be at risk of exercise-related injuries, for example, falls. Whole-body vibration (WBV) is an intervention that helps improve musculoskeletal function and is viable for those with limited mobility. Whether WBV has a dual effect on bone and muscle conditions remains unknown. We aim to assess the evidence of the effects of WBV on bone and muscle parameters concurrently in older people. Under Preferred Reporting Items for Systematic Review and Meta-Analysis guidelines, a systematic literature search was performed in MEDLINE, EMBASE, EMCARE, and the Cochrane Central Registry of Controlled Trials. The main outcomes were changes in bone and muscle parameters. Our meta-analysis showed that WBV does not have significant synergistic effects on measured bone (bone mineral density [BMD] in the hip and lumbar spine) and muscle (lean muscle mass and sit-to-stand time) outcomes, compared to controls (i.e., no WBV included). While there were no significant results, the included studies are limited by small sample size and variable intervention protocols and follow-up periods. Further trials should endeavor to measure both bone and muscle outcomes concurrently with a longer follow-up time. Osteoporosis status in participants must also be considered as it is not yet possible to exclude that WBV may have a significant effect on BMD in people with known osteoporosis. WBV does not appear to simultaneously influence bone and muscle health in older people, and future research is required to establish a regimen that may lead to measurable clinical efficacy.


Geochemical and microbiological evidence for a hydrogen-based, hyperthermophilic subsurface lithoautotrophic microbial ecosystem (HyperSLiME) beneath an active deep-sea hydrothermal field.
Subsurface microbial communities supported by geologically and abiologically derived hydrogen and carbon dioxide from the Earth's interior are of great interest, not only with regard to the nature of primitive life on Earth, but as potential analogs for extraterrestrial life. Here, for the first time, we present geochemical and microbiological evidence pointing to the existence of hyperthermophilic subsurface lithoautotrophic microbial ecosystem (HyperSLiME) dominated by hyperthermophilic methanogens beneath an active deep-sea hydrothermal field in the Central Indian Ridge. Geochemical and isotopic analyses of gaseous components in the hydrothermal fluids revealed heterogeneity of both concentration and carbon isotopic compositions of methane between the main hydrothermal vent (0.08 mM and -13.8 per thousand PDB, respectively) and the adjacent divergent vent site (0.2 mM and -18.5 per thousand PDB, respectively), representing potential subsurface microbial methanogenesis, at least in the divergent vent emitting more 13C-depleted methane. Extremely high abundance of magmatic energy sources such as hydrogen (2.5 mM) in the fluids also encourages a hydrogen-based, lithoautotrophic microbial activity. Both cultivation and cultivation-independent molecular analyses suggested the predominance of Methanococcales members in the superheated hydrothermal emissions and chimney interiors along with the other major microbial components of Thermococcales members. These results imply that a HyperSLiME, consisting of methanogens and fermenters, occurs in this tectonically active subsurface zone, strongly supporting the existence of hydrogen-driven subsurface microbial communities.


The Effectiveness of Mindfulness-Based Stress Reduction on the Psychological Functioning of Healthcare Professionals: a Systematic Review.
Burnout and occupational stress are frequently experienced by healthcare professionals (HCPs). Mindfulness-based stress reduction (MBSR) has been found to improve the psychological health outcomes of HCPs. To date, systematic reviews and meta-analyses have primarily focused upon empirical investigations into the reduction of stress amongst HCPs using MBSR and are limited to empirical studies published before December 2019. This systematic review aimed to update the current evidence base and broaden our understanding of the effectiveness of MBSR on improving the psychological functioning of HCPs. Three electronic databases (Medline, Psych Info and Web of Science) were searched without time frame restrictions. Quantitative studies included randomised controlled trials, clinical controlled trials, pre-post designs and studies with up to a 12-month follow-up period. All studies included in the review employed a MBSR programme, standardised measures of psychological functioning and qualified HCPs as participants. Using PRISMA guidelines thirty studies were included in the review. The reviewed literature suggested that MBSR was effective in reducing HCPs experiences of anxiety, depression and stress. MBSR was also found to be effective in increasing HCP levels of mindfulness and self-compassion. However, MBSR did not appear as effective in reducing burnout or improving resilience amongst HCPs. Abbreviated MBSR programmes were found to be as effective as the traditional 8-week MBSR programmes. MBSR is an effective intervention which can help improve the psychological functioning of HCPs. Recommendations include improving the overall quality of the studies by employing more robust controlled designs with randomisation, increased sample sizes with heterogeneous samples, and making active comparisons between interventions used.


A novel clinically translatable fluorescent nanoparticle for targeted molecular imaging of tumors in living subjects.
The use of quantum dots (QDs) in biomedical research has grown tremendously, yet successful examples of clinical applications are absent due to many clinical concerns. Here, we report on a new type of stable and biocompatible dendron-coated InP/ZnS core/shell QD as a clinically translatable nanoprobe for molecular imaging applications. The QDs (QD710-Dendron) were demonstrated to hold several significant features: near-infrared (NIR) emission, high stability in biological media, suitable size with possible renal clearance, and ability of extravasation. More importantly, a pilot mouse toxicity study confirmed that QD710-Dendron lacks significant toxicity at the doses tested. The acute tumor uptake of QD710-Dendron resulted in good contrast from the surrounding nontumorous tissues, indicating the possibility of passive targeting of the QDs. The highly specific targeting of QD710-Dendron-RGD(2) to integrin α(v)β(3)-positive tumor cells resulted in high tumor uptake and long retention of the nanoprobe at tumor sites. In summary, QD710-Dendron and RGD-modified nanoparticles demonstrate small size, high stability, biocompatibility, favorable in vivo pharmacokinetics, and successful tumor imaging properties. These features satisfy the requirements for clinical translation and should promote efforts to further investigate the possibility of using QD710-Dendron-based nanoprobes in the clinical setting in the near future.


Chick embryo DNA polymerase alpha. Polypeptide components and their microheterogeneity.
DNA polymerase alpha was purified from a chick embryo extract by ammonium sulfate fractionation and successive column chromatographies. The final preparation had a specific enzyme activity of 39,000 units/mg of protein with activated calf thymus DNA as a template.primer. Electrophoresis in nondenaturing polyacrylamide gel showed that the preparation contained two proteins, one of which was associated with DNA polymerase activity. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the electrophoretically pure enzyme gave two clusters of polypeptide bands. One was composed of 3-4 polypeptides with similar electrophoretic mobilities corresponding to Mr = 130,000-155,000, and the other consisted of four distinct polypeptide bands corresponding to Mr = 59,000, 56,000, 54,000, and 51,000. Two-dimensional tryptic peptide mapping analysis of these polypeptides after 125I-iodination indicated that the structures of all four Mr = 51,000-59,000 polypeptides were very similar. In addition, polypeptides in the Mr = 145,000-155,000 region had almost identical structure with those in the Mr = 130,000-140,000 region. No significant structural homology was observed between the high molecular weight polypeptides and low molecular weight polypeptides. These findings indicate that the four polypeptides of Mr = 51,000-59,000 are generated by minor modification(s) of a single polypeptide. Similarly, the heterogeneity of the high molecular weight polypeptides is brought about by minor modification(s). Thus, chick embryo DNA polymerase alpha is basically composed of two kinds of subunit polypeptides.


N-glycan structures of human transferrin produced by Lymantria dispar (gypsy moth) cells using the LdMNPV expression system.
N-glycan structures of recombinant human serum transferrin (hTf) expressed by Lymantria dispar (gypsy moth) 652Y cells were determined. The gene encoding hTf was incorporated into a Lymantria dispar nucleopolyhedrovirus (LdMNPV) under the control of the polyhedrin promoter. This virus was then used to infect Ld652Y cells, and the recombinant protein was harvested at 120 h postinfection. N-glycans were released from the purified recombinant human serum transferrin and derivatized with 2-aminopyridine; the glycan structures were analyzed by a two-dimensional HPLC and MALDI-TOF MS. Structures of 11 glycans (88.8% of total N-glycans) were elucidated. The glycan analysis revealed that the most abundant glycans were Man1-3(+/-Fucalpha6)GlcNAc2 (75.5%) and GlcNAcMan3(+/-Fucalpha6)GlcNAc2 (7.4%). There was only approximately 6% of high-mannose type glycans identified. Nearly half (49.8%) of the total N-glycans contained alpha(1,6)-fucosylation on the Asn-linked GlcNAc residue. However alpha(1,3)-fucosylation on the same GlcNAc, often found in N-glycans produced by other insects and insect cells, was not detected. Inclusion of fetal bovine serum in culture media had little effect on the N-glycan structures of the recombinant human serum transferrin obtained.


Thioredoxin reductase two modes of catalysis have evolved.
Thioredoxin reductase (EC 220.127.116.11) is a widely distributed flavoprotein that catalyzes the NADPH-dependent reduction of thioredoxin. Thioredoxin plays several key roles in maintaining the redox environment of the cell. Like all members of the enzyme family that includes lipoamide dehydrogenase, glutathione reductase and mercuric reductase, thioredoxin reductase contains a redox active disulfide adjacent to the flavin ring. Evolution has produced two forms of thioredoxin reductase, a protein in prokaryotes, archaea and lower eukaryotes having a Mr of 35 000, and a protein in higher eukaryotes having a Mr of 55 000. Reducing equivalents are transferred from the apolar flavin binding site to the protein substrate by distinct mechanisms in the two forms of thioredoxin reductase. In the low Mr enzyme, interconversion between two conformations occurs twice in each catalytic cycle. After reduction of the disulfide by the flavin, the pyridine nucleotide domain must rotate with respect to the flavin domain in order to expose the nascent dithiol for reaction with thioredoxin; this motion repositions the pyridine ring adjacent to the flavin ring. In the high Mr enzyme, a third redox active group shuttles the reducing equivalent from the apolar active site to the protein surface. This group is a second redox active disulfide in thioredoxin reductase from Plasmodium falciparum and a selenenylsulfide in the mammalian enzyme. P. falciparum is the major causative agent of malaria and it is hoped that the chemical difference between the two high Mr forms may be exploited for drug design.


Pushing forces drive the comet-like motility of microtubule arrays in Dictyostelium.
Overexpression of dynein fragments in Dictyostelium induces the movement of the entire interphase microtubule array. Centrosomes in these cells circulate through the cytoplasm at rates between 0.4 and 2.5 microm/s and are trailed by a comet-tail like arrangement of the microtubule array. Previous work suggested that these cells use a dynein-mediated pulling mechanism to generate this dramatic movement and that similar forces are at work to maintain the interphase MTOC position in wild-type cells. In the present study, we address the nature of the forces used to produce microtubule movement. We have used a laser microbeam to sever the connection between the motile centrosomes and trailing microtubules, demonstrating that the major force for such motility results from a pushing on the microtubules. We eliminate the possibility that microtubule assembly/disassembly reactions are significant contributors to this motility and suggest that the cell cortex figures prominently in locating force-producing molecules. Our findings indicate that interphase microtubules in Dictyostelium are subject to both dynein- and kinesin-like forces and that these act in concert to maintain centrosome position in the cell and to support the radial character of the microtubule network.


Erdheim-Chester disease: look it in the eye. An orbital magnetic resonance imaging study.
Erdheim-Chester disease (ECD) is a rare L-group histiocytosis. Orbital involvement is found in a third of cases, but few data are available concerning the radiological features of ECD-related orbital disease (ECD-ROD). Our aim was to characterize the initial radiological phenotype and outcome of patients with ECD-ROD. Initial and follow-up orbital magnetic resonance imaging (MRI) from the patients with histologically proven ECD at a national reference center were reviewed. Pathological orbital findings were recorded for 45 (33%) of the 137 patients included, with bilateral involvement in 38/45 (84%) cases. The mean age (± standard deviation) of these patients was 60 (±11.3) years and 78% were men. Intraconal fat infiltration around the optic nerve sheath adjacent to the eye globe (52%), with intense gadolinium uptake and a fibrous component was the most frequent phenotype described. Optic nerve signal abnormalities were observed in 47% of cases. Two patients had bilateral homogeneous extraocular muscle enlargement suggestive of a myositis-like involvement of ECD-ROD. None had isolated dacryoadenitis but in 17 eyes dacryodenitis was described in association with other types of orbital lesions. Only seven patients (15%) had normal brain MRI findings. ECD-associated paranasal sinus involvement and post-pituitary involvement were detected in 56% and 53% of patients, respectively. A decrease/disappearance of the lesions was observed in 17/24 (71%) of the patients undergoing late (>12 months) followups. Interestingly, ECD-ROD only rarely (7/45; 16%) revealed the disease, with exophthalmos being the most frequently identified feature in this subgroup (3/45; 6%). Even though ECD-ROD can be clinically silent, it comprises a broad array of lesions often resulting in optic nerve signal abnormalities, the functional outcome of which remains to be established. ECD-ROD should thus be assessed initially and subsequently monitored by orbital MRI and ophthalmological follow-up.


Structural plasticity of the circadian timing system. An overview from flies to mammals.
The circadian timing system orchestrates daily variations in physiology and behavior through coordination of multioscillatory cell networks that are highly plastic in responding to environmental changes. Over the last decade, it has become clear that this plasticity involves structural changes and that the changes may be observed not only in central brain regions where the master clock cells reside but also in clock-controlled structures. This review considers experimental data in invertebrate and vertebrate model systems, mainly flies and mammals, illustrating various forms of structural circadian plasticity from cellular to circuit-based levels. It highlights the importance of these plastic events in the functional adaptation of the clock to the changing environment.


[Ludwik Karol Teichmann--a preparator].
Ludwik Karol Teichmann significantly contributed to the creation of modern techniques in the anatomical preparations. He was, next to Joseph Hyrtl, the most versatile among the anatomical preparators in the second half of the 19th century, successfully introducing modifications to existing methods, as well as striving for independent solutions in this field. His precision in performance, transparency and sustainability of the whole brain preparations, excellent osteological preparations, including small bones and cartilage, evoked and still evoke high admiration. He made his name, however, with preparations obtained by means of injection and corrosion techniques. The application of these techniques inthe lymphatic system's study, both the physiologically proper and the pathologically changed, earned Teichmann a permanent position in the history of anatomy. The developed by Teichmann mass for cold injections (the so-called Teichmann's cold mass) revolutionized the macro-and microscopic preparatory of that time, thus opening great new research perspectives still widely used during the interwar period.


Metastasis from breast cancer to an endometrial polyp; treatment options and follow-up. Report of a case and review of the literature.
The female genital tract is rarely involved by metastatic tumors. The most common anatomic locations are the ovaries and the vagina. A case is presented of metastatic breast carcinoma to the vulva and endometrial polyp, both exceptional. We report the case of an 83-year-old female who presented with vaginal bleeding. Lobular breast carcinoma was diagnosed earlier and during follow-up vulvar metastasis was detected. Hysteroscopic examination because of postmenopausal bleeding revealed an endometrial polyp which was resected. The morphology and immunohistochemistry of the polyp were consistent with lobular breast cancer: metastatic breast cancer to an endometrial polyp. After reviewing the literature 15 cases of metastatic breast carcinoma to endometrial polyps have been reported. The clinical presentation and course, risk factors, treatment and follow-up are discussed. Metastasis of a breast carcinoma to the vulva and an endometrial polyp are extremely rare, but clinicians should be aware of both phenomena.


The importance of the family history in caring for families with long QT syndrome and dilated cardiomyopathy.
In potentially inherited cardiac diseases, the family history is of great importance. We looked at the way cardiologists take a family history in patients with idiopathic dilated cardiomyopathy (DCM) or long QT syndrome (LQTS) and whether this led to screening of relatives or other follow-up. We performed retrospective cross-sectional analyses of adult index patients with DCM or LQTS in a general hospital (GH) or a University Medical Center (UMC). We identified 82 index patients with DCM (34 GH; 48 UMC) and 20 with LQTS (all UMC) between 1996 and 2005. Mean follow-up was 58 months. A family history was recorded in 90% of both LQTS and DCM patients most of the cases restricted to first-degree family members. The genetic aspects, counseling and screening of family members was discussed significantly more often with LQTS than DCM patients (all P < 0.05). Also follow-up (screening of family members, DNA analysis and referral) was performed significantly more often in LQTS than DCM patients. Cardiologists in the UMC referred DCM index patients for genetic counseling more often than those in the GH (25% vs. 6%; P < 0.05). Only a few index patients with DCM were referred to a clinical genetics department. One-third of DCM cases and nearly all LQTS cases are familial. Since early recognition and treatment may reduce morbidity and mortality we recommend cardiologists take a more thorough family history and always consider referring to a clinical genetics department in such index patients.


[Acute pancreatitis and afferent loop syndrome].
A new case of afferent loop obstruction presenting as acute pancreatitis is described. It is an uncommon manifestation of the afferent loop syndrome: less than 20 cases found in the literature. The diagnosis of afferent loop obstruction should be suspected in any patient with acute pancreatitis and a previous Billroth II gastrectomy. Duodenopancreatic reflux seems to play an important role in the pathogenesis of acute pancreatitis in this situation. The "closed duodenal loop" experimental model of acute pancreatitis simulates quite closely this clinical situation. The physiopathological principles, diagnosis and treatment of this syndrome is reviewed.


Maternal-infant separation and breast-feeding. The return to work or school.
Breast feeding is now practiced by more than 50% of new mothers, many of whom return to work or school within six to eight weeks postpartum. Their breast-feeding experience need not, and should not, be terminated because of this temporary, though regular, separation from their infants. The obstetrician has a role in facilitating continued breast-feeding after the return to work or school. Counseling these patients should include a discussion of hand expression and breast pumping, the means of storing human milk, infant changes related to regular maternal absences and avoiding exhaustion.


Role of Iron on Physical and Mechanical Properties of Brushite Cements, and Interaction with Human Dental Pulp Stem Cells.
Improving the physical, mechanical and biological properties of brushite cements (BrC) is of a great interest for using them in bone and dental tissue engineering applications. The objective of this study was to incorporate iron (Fe) at different concentrations (0.25, 0.50, and 1.00 wt.%) to BrC and study the role of Fe on phase composition, setting time, compressive strength, and interaction with human dental pulp stem cells (hDPSCs). Results showed that increase in Fe concentration increases the β-tricalcium phosphate (β-TCP)/ dicalcium phosphate dihydrate (DCPD) ratio and prolongs the initial and final setting time due to effective role of Fe on stabilizing the β-TCP crystal structure and retarding its dissolution kinetic, in a dose dependent manner where the highest setting time was recorded for 1.00 wt.% Fe-BrC sample. Addition of low concentrations of Fe (0.25 and 0.50 wt.%) did not have adverse effect on compressive strength and strength was in the range of 5.7-7.05 (±~1.4) MPa; however, presence of 1.00 wt.% Fe decreases the strength of BrC from 7.05 ± 1.57 MPa to 3.12 ± 1.06 MPa. Interaction between the BrCs and hDPSCs was evaluated by cell proliferation assay, scanning electron microscopy, and live/dead staining. Low concentrations of 0.25, and 0.50 wt.% of Fe did not have any adverse effect on cell attachment and proliferation; while significant decrease in cellular activity was evident in BrC samples doped with 1.00 wt. %. Together, these data show that low concentrations of Fe (equal or less than 0.50 wt. %) can be safely added to BrC without any adverse effect on physical, mechanical and biological properties in presence of hDPSCs.


Deep mitochondrial DNA phylogeographic divergence in the threatened aoudad Ammotragus lervia (Bovidae, Caprini).
The aoudad or Barbary sheep (Ammotragus lervia) is a threatened ungulate emblematic of North Africa, whose population structure and subspecific taxonomy have not been examined genetically. This knowledge is essential and urgently needed to inform ongoing conservation and management efforts. We analysed the mitochondrial cytochrome b gene and four nuclear genes (casein kappa, spectrin beta nonerythrocytic 1, thyroglobulin, thyrotropin subunit beta) for the first phylogeographic survey of the aoudad, and uncovered a deep Mediterranean-Saharan mitochondrial split separating two highly distinct evolutionary lineages. Their level of divergence is greater than or comparable to those observed between several pairs of congeneric species of different caprine genera. The split was estimated to have occurred in the Early Pleistocene, about 1.3 million years ago. None of the four nuclear genes surveyed, chosen because they have been used in phylogeographic and species-level phylogenetic studies of bovids, allowed us to detect, likely due to their slow evolutionary rate, the substantial and geographically coherent subdivision revealed by mitochondrial DNA. This study is evidence and testament to the ability of mitochondrial DNA, probably unrivalled by any other single-locus marker, as an exploratory tool for investigating population genealogy and history and identifying potential evolutionarily significant units for conservation in animals.


Improved methods for making and using glass knives.
We have observed over time that the right side of a glass knife is the optimal cutting edge for microtomy if the counterpiece (heel opposite the edge) is controlled within 1 mm. The right cutting edge has been considered the "saw toothed" side and has not been used for ultrathin sectioning. We have observed that the right cutting edge is sharper and more durable than the left. Light and scanning electron microscopy were used to observe the cutting edge, and transmission electron microscopy was used to examine semithin and ultrathin sections of animal and plant tissues cut by the right and left sides of the cutting edge. The results indicate that the cutting edge becomes sharper and more durable from left to right. Both the quality and efficiency of ultrathin sectioning is improved by using the right cutting edge.


Ultrastructural Analysis of Incinerated Teeth by Scanning Electron Microscope - A Short Study.
In forensic context precise knowledge on physical and histological changes of teeth subjected to high temperatures is of great importance. Preserving fragile incinerated teeth for physical, histological and ultra structural examinations is essential in fire investigations involving the origin of fire, its cause as well as the identification of victims which relies on a thorough understanding of the structural changes in dental tissues subjected to heat. The study was conducted to evaluate the physical and ultrastructural changes seen in freshly extracted teeth when subjected to gradual heating at different temperatures using Scanning Electron Microscope (SEM). Freshly extracted teeth collected from subjects of different age groups were subjected to different temperatures using laboratory furnace and findings were correlated to the temperature. The study was conducted on 60 healthy freshly extracted teeth belonging to age group between 20-30 years. Group A comprised of control group which included teeth that were not subjected to heat whereas Group B, C and D comprised of teeth that were subjected to different temperatures i.e., 100(o)C, 300(o)C and 600(o)C respectively for a time of fifteen minutes in laboratory furnace, after which they were processed for SEM examination. Each group included 15 teeth; 5 anteriors, 5 premolars and 5 molars. Examination under SEM revealed definite ultra structural changes which were explicitly seen at particular temperatures (100(o)C, 300(o)C and 600(o)C). The samples showed cracks and charring of the tooth structure with ultra structural findings such as pebbles, granules, dots on enamel surface; and soap bubble pattern, honey comb pattern and snail track pattern on cementum surface. Because of the consistency of morphological changes and the ultra structural patterns at various temperatures, evaluation of incinerated dental remains using SEM can provide additional investigative avenues in victim identification and in the areas of forensic fire investigations.


Socio-economic status and overall and cause-specific mortality in Sweden.
Previous studies have reported discrepancies in cause-specific mortality among groups of individuals with different socio-economic status. However, most of the studies were limited by the specificity of the investigated populations and the broad definitions of the causes of death. The aim of the present population-based study was to explore the dependence of disease specific mortalities on the socio-economic status in Sweden, a country with universal health care. Another aim was to investigate possible gender differences. Using the 2006 update of the Swedish Family-Cancer Database, we identified over 2 million individuals with socio-economic data recorded in the 1960 national census. The association between mortality and socio-economic status was investigated by Cox's proportional hazards models taking into account the age, time period and residential area in both men and women, and additionally parity and age at first birth in women. We observed significant associations between socio-economic status and mortality due to cardiovascular diseases, respiratory diseases, to cancer and to endocrine, nutritional and metabolic diseases. The influence of socio-economic status on female breast cancer was markedly specific: women with a higher socio-economic status showed increased mortality due to breast cancer. Even in Sweden, a country where health care is universally provided, higher socio-economic status is associated with decreased overall and cause-specific mortalities. Comparison of mortality among female and male socio-economic groups may provide valuable insights into the underlying causes of socio-economic inequalities in length of life.


Effects of K-351 on adrenergically induced renin release and renal vasoconstriction in anesthetized dogs.
Effect of 3-4-dihydro-8-(2-hydroxy-3-isopropylaminopropoxy)-3-nitroxy-2H-1-b enzopyran (K-351) infused into the renal artery on renin release during graded renal nerve stimulation (RNS) was investigated in pentobarbital-anesthetized dogs. K-351 (20 micrograms/min) produced significant suppression of the RNS-induced renin release; the renin secretion responses to RNS at lower frequencies (0.5 and 1 Hz) were almost abolished, and those at the highest frequency (3 Hz) were attenuated. K-351 also suppressed an increase in renal vascular resistance during RNS at 3 Hz. The same extent of inhibition in the renin secretion response to RNS was also obtained during infusion of DL-propranolol (100 micrograms/min). Inhibitory effect of K-351, prazosin, or phentolamine on the renal vasoconstriction induced by RNS and norepinephrine (NE) injected into the renal artery, an approximately 50% reduction in renal blood flow, was also assessed. K-351 and prazosin exerted a greater inhibitory effect on RNS-than on NE-induced vasoconstriction, and the opposite was true of phentolamine. The potency of K-351 in reducing the vasoconstriction due to RNS or NE was roughly estimated to be 10-30 times less than that of prazosin. These results suggest that K-351 shares beta- and alpha-adrenoceptor blocking properties, which effectively contribute to the suppression of adrenergically induced renin release and renal vasoconstriction.


Validation of the Empathy Quotient in Mainland China.
This research aimed to validate a simplified Chinese version of the Empathy Quotient (EQ; 60 items) for use with Mainland Chinese people. The original English version of the EQ was translated into simplified Chinese. Through an online survey, 588 Mainland Chinese participants completed the EQ and 3 other questionnaires: the Interpersonal Reactivity Index (IRI), the Autism-Spectrum Quotient (AQ), and the 20-item Toronto Alexithymia Scale (TAS-20). Thirty-five participants completed retesting of the EQ 3 to 4 weeks later. Sex differences on the EQ scores and psychometric properties of the EQ items were examined. Confirmatory factor analysis suggested that an EQ 15-item structural model fitted the data quite well. Self-report empathy, as assessed by the current simplified Chinese version of the EQ, appeared to relate to participants' autistic and alexithymic traits but not sex.


Attention switching during scene perception: how goals influence the time course of eye movements across advertisements.
Eye movements across advertisements express a temporal pattern of bursts of respectively relatively short and long saccades, and this pattern is systematically influenced by activated scene perception goals. This was revealed by a continuous-time hidden Markov model applied to eye movements of 220 participants exposed to 17 ads under a free-viewing condition, and a scene-learning goal (ad memorization), a scene-evaluation goal (ad appreciation), a target-learning goal (product learning), or a target-evaluation goal (product evaluation). The model reflects how attention switches between two states--local and global--expressed in saccades of shorter and longer amplitude on a spatial grid with 48 cells overlaid on the ads. During the 5- to 6-s duration of self-controlled exposure to ads in the magazine context, attention predominantly started in the local state and ended in the global state, and rapidly switched about 5 times between states. The duration of the local attention state was much longer than the duration of the global state. Goals affected the frequency of switching between attention states and the duration of the local, but not of the global, state.


Electrocatalysis of formic acid on palladium and platinum surfaces: from fundamental mechanisms to fuel cell applications.
Formic acid as a natural biomass and a CO2 reduction product has attracted considerable interest in renewable energy exploitation, serving as both a promising candidate for chemical hydrogen storage material and a direct fuel for low temperature liquid fed fuel cells. In addition to its chemical dehydrogenation, formic acid oxidation (FAO) is a model reaction in the study of electrocatalysis of C1 molecules and the anode reaction in direct formic acid fuel cells (DFAFCs). Thanks to a deeper mechanistic understanding of FAO on Pt and Pd surfaces brought about by recent advances in the fundamental investigations, the "synthesis-by-design" concept has become a mainstream idea to attain high-performance Pt- and Pd-based nanocatalysts. As a result, a large number of efficient nanocatalysts have been obtained through different synthesis strategies by tailoring geometric and electronic structures of the two primary catalytic metals. In this paper, we provide a brief overview of recent progress in the mechanistic studies of FAO, the synthesis of novel Pd- and Pt-based nanocatalysts as well as their practical applications in DFAFCs with a focus on discussing studies significantly contributing to these areas in the past five years.


Identification of highly potent and selective Cdc25 protein phosphatases inhibitors from miniaturization click-chemistry-based combinatorial libraries.
Cell division cycle 25 (Cdc25) protein phosphatases play key roles in the transition between the cell cycle phases and their association with various cancers has been widely proven, which makes them ideal targets for anti-cancer treatment. Though several Cdc25 inhibitors have been developed, most of them displayed low activity and poor subtype selectivity. Therefore, it is extremely important to discover novel small molecule inhibitors with potent activities and significant selectivity for Cdc25 subtypes, not only served as drugs to treat cancer but also to probe its mechanism in transitions. In this study, miniaturized parallel click chemistry synthesis via CuAAC reaction followed by in situ biological screening were used to discover selective Cdc25 inhibitors. The bioassay results showed that compound M2N12 proved to be the most potent Cdc25 inhibitor, which also act as a highly selective Cdc25C inhibitor and was about 9-fold potent than that of NSC 663284. Moreover, M2N12 showed remarkable anti-growth activity against the KB-VIN cell line, equivalent to that of PXL and NSC 663284. An all-atom molecular dynamics (MD) simulation approach was further employed to probe the significant selectivity of M2N12 for Cdc25C relative to its structural homologs Cdc25A and Cdc25B. Overall, above results make M2N12 a promising lead compound for further investigation and structural modification.


Fusion dsRNA in targeting salivary protein genes enhance the RNAi-based aphid control.
RNA interference (RNAi) is a promising tool for pest control and relies on sequence-specific gene silencing. Salivary proteins are cooperatively secreted into plants to guarantee the feeding of aphids; thus they have potential to develop as selective targets for RNAi-based pest control strategy. For this purpose, we firstly analyzed 18 salivary proteomes of various aphid species, and these salivary proteins can be mainly categorized into seven functional groups. Secondly, we created a work-flow for fusion dsRNA design that can target multiple genes but were selectively safe to beneficial insects. Based on this approach, seven fusion dsRNAs were designed to feed the green peach aphid, which induced a significant reduction in aphid fitness. Among them, ingestion of dsperoxidase induced the highest mortality in aphids, which was also significantly higher than that of traditional dsRNAs in targeting three peroxidases separately. In addition, dsperoxidase-fed green peach aphids triggered the highest H2O2 content of host plants as well as the attraction to natural enemies (ladybeetle and parasitic wasp) but repellent to other control aphids. Our results indicate that the fusion dsRNA design approach can improve aphid control capacity, and the fusion dsRNA targeting salivary protein-encoding genes can enhance the direct and indirect defenses of host plants, thus providing a new strategy for RNAi-based aphid control.


Being in the situation of a significant other to a person with inoperable lung cancer.
Little is known about how significant others experience their own situation when a family member becomes seriously ill. To illuminate the meanings of significant others' experiences of their situation after a family member had been diagnosed with inoperable lung cancer, we analyzed narratives from 12 significant others, using a phenomenological hermeneutic approach. Findings revealed 4 themes: feeling dislocated in life, being in an altered relationship, being in a struggle, and feeling secure. When reflecting on these findings, we considered the transition theories and works of Ricoeur and Lögstrup. The significant others' experiences indicated a transition process because of the changes brought about by the diagnosis of lung cancer and a struggle to endure and overcome difficulties and distress on the way to regaining a smooth functioning life. Furthermore, the diagnosis of lung cancer had altered the relationship toward the next of kin and meant not only feelings of increased closeness but also loss of intimacy and reciprocity. The significant others suffered in this process of transition, and healthcare professionals, who recognize the World Health Organization's definition of palliative care, may consider the findings of this study useful when reflecting on their care of families.


Expression by human neuroblastoma cells of an antigen recognized by naturally occurring mouse anti-brain autoantibody.
A number of human tumor cell lines of both neural and nonneural origin have been assayed for the expression of an interspecies brain antigen (mouse brain antigen 2), detected by naturally occurring antibodies in normal mouse sera. These experiments indicate that mouse brain antigen 2 is present on four human neuroblastoma cell lines but not on other human tumor cell lines tested, including four glial cell lines and a retinoblastoma. These findings demonstrate the value of naturally occurring antibodies in normal mouse sera for the detection and serological classification of human tumor antigens and indicate that considerable caution should be used in attempts to distinguish tumor-specific and tissue-specific antigen expression on human neuroblastoma cells.


Folate and homocysteine in the cerebrospinal fluid of patients with Alzheimer's disease or dementia: a case control study.
Amyloid deposition in the brain is an early event in Alzheimer's disease (AD), but a dysfunction of the blood-brain barrier or a disturbance in the metabolism of folate and homocysteine (Hcy) may affect the development of dementia. We investigated if the concentrations of folate and Hcy would be modified in cerebrospinal fluid (CSF) of clinically diagnosed AD patients. We included 70 AD patients, 33 patients with another type of dementia (nAD) and 30 age-matched control subjects. Plasma Hcy was assayed as well as Hcy, folate, Aβ1-42 and T-tau in CSF. We used ANOVAs for comparison between groups, and then pairwise comparisons by Wilcoxon tests with Bonferroni-corrected p values. Correlations were tested with the Spearman's rank test. Levels of Aβ1-42, T-tau and folates in CSF were significantly different between groups, but not Hcy. In addition, the average folate in CSF was lower in AD patients compared with controls (18.7 ± 2.4 vs. 20.3 ± 1.7 nmol/l, Bonferroni-corrected p value < 0.02). There was no correlation between Aβ1-42 or T-tau and folate or Hcy in CSF, regardless of the group. In the AD group, there was a significant inverse correlation between Hcy and folate in CSF (ρ = -0.63, p < 0.0001), whereas in the nAD group, a significant correlation was found for Hcy between plasma and CSF (ρ = 0.59, p < 0.0005). The concentration of folate in CSF was found to be decreased in AD patients. These findings support the hypothesis of a possible role of folate in the onset or worsening of AD.


Towards an integrative understanding of cancer mechanobiology: calcium, YAP, and microRNA under biophysical forces.
An increasing number of studies have demonstrated the significant roles of the interplay between microenvironmental mechanics in tissues and biochemical-genetic activities in resident tumor cells at different stages of tumor progression. Mediated by molecular mechano-sensors or -transducers, biomechanical cues in tissue microenvironments are transmitted into the tumor cells and regulate biochemical responses and gene expression through mechanotransduction processes. However, the molecular interplay between the mechanotransduction processes and intracellular biochemical signaling pathways remains elusive. This paper reviews the recent advances in understanding the crosstalk between biomechanical cues and three critical biochemical effectors during tumor progression: calcium ions (Ca2+), yes-associated protein (YAP), and microRNAs (miRNAs). We address the molecular mechanisms underpinning the interplay between the mechanotransduction pathways and each of the three effectors. Furthermore, we discuss the functional interactions among the three effectors in the context of soft matter and mechanobiology. We conclude by proposing future directions on studying the tumor mechanobiology that can employ Ca2+, YAP, and miRNAs as novel strategies for cancer mechanotheraputics. This framework has the potential to bring insights into the development of novel next-generation cancer therapies to suppress and treat tumors.


Coordination of three signaling enzymes by AKAP79, a mammalian scaffold protein.
Multivalent binding proteins, such as the yeast scaffold protein Sterile-5, coordinate the location of kinases by serving as platforms for the assembly of signaling units. Similarly, in mammalian cells the cyclic adenosine 3',5'-monophosphate-dependent protein kinase (PKA) and phosphatase 2B [calcineurin (CaN)] are complexed by an A kinase anchoring protein, AKAP79. Deletion analysis and binding studies demonstrate that a third enzyme, protein kinase C (PKC), binds AKAP79 at a site distinct from those bound by PKA or CaN. The subcellular distributions of PKC and AKAP79 were similar in neurons. Thus, AKAP79 appears to function as a scaffold protein for three multifunctional enzymes.


Estimated Cost Effectiveness of Influenza Vaccination for Emergency Medical Services Professionals.
Because of their frequent contact with compromised patients, vaccination against influenza is recommended for all healthcare workers. Recent studies suggest that vaccination decreases influenza transmission to patients and reduces worker illness and absenteeism. However, few emergency medical services (EMS) agencies provide annual vaccination, and the vaccination rate among EMS personnel remains low. Reticence among EMS agencies to provide influenza vaccination to their employees may be due in part to the unknown fiscal consequences of implementing a vaccination program. In this study, we sought to estimate the cost effectiveness of an employer-provided influenza vaccination program for EMS personnel. Using data from published reports on influenza vaccination, we developed a cost-effectiveness model of vaccination for a hypothesized EMS system of 100 employees. Model inputs included vaccination costs, vaccination rate, infection rate, costs associated with absenteeism, lost productivity due to working while ill (presenteeism), and medical care for treating illness. To assess the robustness of the model we performed a series of sensitivity analyses on the input variables. The proportion of employees contracting influenza or influenza-like illness (ILI) was estimated at 19% among vaccinated employees compared to 26% among non-vaccinated employees. The costs of the vaccine, consumables, and employee time for vaccination totaled $44.19 per vaccinated employee, with a total system cost of $4,419. Compared to no vaccination, a mandatory vaccination program would save $20,745 in lost productivity and medical costs, or $16,325 in net savings after accounting for vaccination costs. The savings were 3.7 times the cost of the vaccination program and were derived from avoided absenteeism ($7,988), avoided presenteeism productivity losses ($10,303), and avoided medical costs of treating employees with influenza/ILI ($2,454). Through sensitivity analyses the model was verified to be robust across a wide range of input variable assumptions. The net monetary benefits were positive across all ranges of input assumptions, but cost savings were most sensitive to the vaccination uptake rate, ILI rate, and presenteeism productivity losses. This cost-effectiveness analysis suggests that an employer-provided influenza vaccination program is a financially favorable strategy for reducing costs associated with influenza/ILI employee absenteeism, presenteeism, and medical care.


Multicenter trial comparing tramadol and morphine for pain after abdominal surgery.
Seventy patients (40 male, 30 female), mean (SD) age 60.8 +/- 13.7 years were treated with parenteral morphine (10 mg/1 ml ampul) or tramadol (100 mg/2 ml ampul) to verify their analgesic effects in pain following abdominal surgery. The multicenter trial followed an open, controlled experimental design between patients, randomized within the centers. The drugs were given by intramuscular injection, as requested by patients, starting in the postoperative period when pain was more than 70 mm, assessed on a visual analog scale. Patients were allowed up to six ampuls of tramadol or morphine in the 24-h trial but in the first 4 h, if they asked for supplementary analgesic, only diclofenac (75 mg in a 3-ml ampul) was allowed. Both test drugs gave rapid and constant pain relief. After the first dose, pain intensity was reduced 36.2% with tramadol, and 51% with morphine; the pain-free interval was similar for both treatments. The quality of sleep and the number of hours of sleep the night after surgery were similar for both groups. Tramadol was tolerated better, giving rise to no untoward reactions; with morphine there was one case of mild respiratory depression. In abdominal surgery, therefore, tramadol given by intramuscular injection has postoperative analgesic activity similar to morphine, but is better tolerated.


Assessment of factors associated with functional status in 60 years-old and older adults in Bogotá, Colombia.
Functional impairment produces a wide range of negative effects such as difficulty in mobility, social isolation, decreased quality of life, disability and institutionalization. Thus, functional status measurement is a marker of social wellbeing. To determine and characterize the socio-demographic factors and health conditions related to functional impairment in older adults in Bogotá, Colombia. Data was collected from the SABE Bogotá Study. Functional status was assessed using the Barthel and Lawton scales. Independent variables included socio-demographic factors, comorbidities, anthropometric measurements and physical activity (by tertiles). Bivariate analyzes were performed, and multivariate results were obtained using linear regression models. There were 2,000 participants aged 60 years and older, with a mean age of 71.2 ± 8 years. Younger age (standardized beta = -0.15, p<0.01), fewer medications (beta= -0.13, p<0.01), higher MMSE score (beta = 0.3, p< 0.01), higher level of physical activity (middle beta tertile = 0.18, p<0.01, and higher beta tertile= 0.18, p<0.01vs lower tertile) and better performance in the handgrip test (beta= 0.10, p<0.01) were associated with better function in daily life instrumental activities. Similarly, higher MMSE score (beta= 0.3, p<0.01) and better performance in the handgrip test (beta= 0.07, p= 0.02) were associated with better function in daily life basic activities. We found several factors related with functional impairment, which are likely to be modified to reduce dependence in this population.


Optic neuritis and computed tomography.
A forty year old Lebanese man presented to Royal Prince Alfred Hospital with unilateral visual loss and disc oedema. Computed Tomography (CT) of his orbit demonstrated diffuse thickening and tortuosity consistent with an optic nerve tumour. Surgical exploration of the orbit was averted when the patient reported subjective visual improvement. Two months later, CT demonstrated an optic nerve which was well within normal limits.


N-[6-(4-butanoyl-5-methyl-1H-pyrazol-1-yl)pyridazin-3-yl]-5-chloro-1-[2-(4-methylpiperazin-1-yl)-2-oxoethyl]-1H-indole-3-carboxamide (SAR216471), a novel intravenous and oral, reversible, and directly acting P2Y12 antagonist.
In the search of a potential backup for clopidogrel, we have initiated a HTS campaign designed to identify novel reversible P2Y12 antagonists. Starting from a hit with low micromolar binding activity, we report here the main steps of the optimization process leading to the identification of the preclinical candidate SAR216471. It is a potent, highly selective, and reversible P2Y12 receptor antagonist and by far the most potent inhibitor of ADP-induced platelet aggregation among the P2Y12 antagonists described in the literature. SAR216471 displays potent in vivo antiplatelet and antithrombotic activities and has the potential to differentiate from other antiplatelet agents.