Quantitative Evaluation of the Community Research Fellows Training Program.
The community research fellows training (CRFT) program is a community-based participatory research (CBPR) initiative for the St. Louis area. This 15-week program, based on a Master in Public Health curriculum, was implemented by the Division of Public Health Sciences at Washington University School of Medicine and the Siteman Cancer Center. We measure the knowledge gained by participants and evaluate participant and faculty satisfaction of the CRFT program both in terms of meeting learning objectives and actively engaging the community in the research process. We conducted analyses on 44 community members who participated in the CRFT program and completed the baseline and follow-up knowledge assessments. Knowledge gain is measured by a baseline and follow-up assessment given at the first and final session. Additionally, pre- and post-tests are given after the first 12 sessions. To measure satisfaction, program evaluations are completed by both the participants and faculty after each topic. Mid-way through the program, a mid-term evaluation was administered to assess the program's community engagement. We analyzed the results from the assessments, pre- and post-tests, and evaluations. The CRFT participants' knowledge increased at follow-up as compared with baseline on average by a 16.5 point difference (p < 0.0001). Post-test scores were higher than pre-test scores for 11 of the 12 sessions. Both participants and faculty enjoyed the training and rated all session well. The CRFT program was successful in increasing community knowledge, participant satisfaction, and faculty satisfaction. This success has enhanced the infrastructure for CBPR as well as led to CBPR pilot projects that address health disparities in the St. Louis Greater Metropolitan Area.


Cervical cancer: a missed health priority in Tanzania.
Cervical cancer is a malignant neoplasm of the cervix uteri. It is the second commonest cancer in women worldwide and is among the largest causes of global cancer mortality. Human papilloma virus (HPV) which is transmitted sexually, particularly subtypes 16 and 18 are responsible for causing majority of cervical cancer cases worldwide. The disease is one of the most preventable and curable carcinomas if detected and treated at an early enough stage. The effective prevention and control of the disease depends on, among others, the effective screening program coupled with knowledge and awareness of women population on the disease's risk factors and available screening services. Pap-test screening that is widely used in Western countries and proved to be effective in reducing the incidence of cervical cancer is not so popular in Tanzania, and so is HPV vaccine. This review examines the literatures on cervical cancer situation and explores various evidence-based cost-effective strategies and approaches that could be employed to confront the rising cervical cancer burden in the country.


Two types of calcium channels in single smooth muscle cells from rabbit ear artery studied with whole-cell and single-channel recordings.
Freshly dispersed rabbit ear artery cells were studied using patch-clamp techniques to measure membrane currents in whole cells and single-channel currents in membrane patches. Whole-cell calcium currents recorded at physiologic extracellular calcium concentrations were small (approximately 10 pA). Recordings with 110 mM external barium gave much larger currents and revealed two current components with properties similar to those in other vascular smooth muscle preparations and to the calcium currents designated as T and L in heart cells and neurons. T current was activated with weak depolarizations and inactivated rapidly, while L current was activated with relatively strong depolarizations and inactivated more slowly. L current was increased by dihydropyridine agonists and decreased by dihydropyridine antagonists, while T current was unaffected. Recordings from cell-attached and outside-out membrane patches with 100 mM external barium showed unitary calcium channel currents with conductances of 8 and 25 pS. The small conductance channels had kinetic properties that accounted for T current in the whole-cell recordings, while the 25-pS channels showed the voltage dependence, the time dependence, and the dihydropyridine sensitivity expected for L-type channels. We conclude that vascular smooth muscle cells contain two types of calcium channels with properties very similar to those described for T- and L-type calcium channels in other cell types; the L current appears to be the predominant current component in whole-cell recordings.


Orbicularis Oculi Muscle Reinnervation Confers Corneal Protective Advantages over Static Interventions Alone in the Subacute Facial Palsy Patient.
Corneal protection is a priority in flaccid facial palsy patients. Denervation of the orbicularis oculi muscle results in weak palpebral closure and predisposes patients to severe corneal sequelae. While periorbital static procedures enhance corneal coverage in repose, voluntary closure is only regained through dynamic reinnervation of the muscle. This study aims to elucidate the added effect of dynamic reinnervation of the orbicularis oculi muscle on long-term corneal integrity as well as on dynamic closure of the palpebral aperture. Retrospective review was performed on two groups of complete palsy patients: those who received solely periorbital static procedures and those who underwent concomitant orbicularis oculi muscle reinnervation and static lid procedures. Only patients with complete ophthalmic examinations were included. Corneal punctate epithelial erosions in addition to static and dynamic palpebral measurements were serially assessed preoperatively and postoperatively. Of 272 facial palsy patients, 26 fit the inclusion criteria. Eleven patients underwent combined muscle reinnervation involving facial-to-masseteric nerve coaptation in addition to static eye procedures, and 15 patients underwent solely static interventions. Analysis revealed a 65.3 percent lower mean punctate epithelial erosion score in reinnervation patients as compared with static patients when evaluated at more than 9 months postoperatively (p < 0.01). Reinnervation patients were also found to have 25.3 percent greater palpebral aperture closure (p < 0.05) and 32.8 percent higher closure velocity (p < 0.01) compared with static patients. In patients with subacute facial palsy, dynamic reanimation of the orbicularis oculi muscle with concomitant static interventions provides lasting corneal protection not seen in patients who receive solely static interventions. Therapeutic, III.


Kaposi's sarcoma in an HIV-positive person successfully treated with paclitaxel.
Epidemic Kaposi's sarcoma is one of the malignant neoplasms, which can develop in HIV-infected patients. Although the prevalence of HIV infection is reported to be high in Asian countries, Kaposi's sarcoma is rarely reported. We report a case of Kaposi's sarcoma involving the skin and oral mucosa along with extensive bilateral lymphedema of lower extremities, treated successfully with paclitaxel and antiretrovirals.


Provision of individualized information to men and their partners to facilitate treatment decision making in prostate cancer.
To determine if providing individualized information to men who are newly diagnosed with prostate cancer and their partners would lower their levels of psychological distress and enable them to become more active participants in treatment decision making. Quasiexperimental, one group, pretest/post-test. The Prostate Centre at Vancouver General Hospital in British Columbia, Canada. Convenience sample of 74 couples. 73 men had early-stage prostate cancer. Mean age of the men was 62.2 years, and mean age of the partners was 58.1 years. The majority (> 50%) had received their high school diplomas. Respondents completed measures of decision preferences and psychological distress at the time of diagnosis and four months later. All participants used a computer to identify their information and decision preferences. Computer-generated, graphic printouts were used to guide the information counseling session. Patients reported assuming a more active role in medical decision making than originally intended, partners assumed a more passive role in decision making than originally intended, and all participants had lower levels of psychological distress at four months. Evidence supports the need to provide informational support to couples at the prostate cancer diagnosis to facilitate treatment decision making and lower levels of psychological distress. Future research is needed to evaluate this type of approach in the context of a randomized clinical trial design. The personalized, computer-graphic printouts can provide clinicians with an innovative method of guiding information counseling and providing decisional support to men with prostate cancer and their partners.


Brain activity associated with expectancy-enhanced placebo analgesia as measured by functional magnetic resonance imaging.
In this study, a well established expectancy manipulation model was combined with a novel placebo intervention, a validated sham acupuncture needle, to investigate the brain network involved in placebo analgesia. Sixteen subjects completed the experiment. We found that after placebo acupuncture treatment, subjective pain rating reduction (pre minus post) on the placebo-treated side was significantly greater than on the control side. When we calculated the contrast that subtracts the functional magnetic resonance imaging (fMRI) signal difference between post-treatment and pretreatment during pain application on placebo side from the same difference on control side [e.g., placebo (post - pre) - control (post - pre)], significant differences were observed in the bilateral rostral anterior cingulate cortex (rACC), lateral prefrontal cortex, right anterior insula, supramarginal gyrus, and left inferior parietal lobule. The simple regression (correlation) analysis between each subject's fMRI signal difference of post-treatment and pretreatment difference on placebo and control side and the corresponding subjective pain rating difference showed that significant negative correlation was observed in the bilateral lateral/orbital prefrontal cortex, rACC, cerebellum, right fusiform, parahippocampus, and pons. These results are different from a previous study that found decreased activity in pain-sensitive regions such as the thalamus, insula, and ACC when comparing the response to noxious stimuli applied to control and placebo cream-treated areas of the skin. Our results suggest that placebo analgesia may be configured through multiple brain pathways and mechanisms.


Directiveness in teachers' language input to toddlers and preschoolers in day care.
Five subtypes of directiveness were examined in the interactions of day care teachers with toddler and preschooler groups. The instructional context (book reading, play dough) yielded significant differences across all five subtypes of directiveness, indicating that these two activities elicited different types of teacher-child discourse. Book reading was characterized by significantly more behavior and response control and less conversation control in comparison with the play-dough activity. Correlations between teachers' directiveness and child language productivity indicated that behavior control and turn-taking control were associated with low levels of productivity, whereas conversation control was associated with the highest levels of productivity. The results of this study confirm that instructional context is an important mediator of teachers' directiveness and suggest that subtypes of directiveness have differential effects on child language output.


Molecular basis of gain-of-function LEOPARD syndrome-associated SHP2 mutations.
The Src homology 2 (SH2) domain-containing protein tyrosine phosphatase 2 (SHP2) is a critical signal transducer downstream of growth factors that promotes the activation of the RAS-ERK1/2 cascade. In its basal state, SHP2 exists in an autoinhibited closed conformation because of an intramolecular interaction between its N-SH2 and protein tyrosine phosphatase (PTP) domains. Binding to pTyr ligands present on growth factor receptors and adaptor proteins with its N-SH2 domain localizes SHP2 to its substrates and frees the active site from allosteric inhibition. Germline mutations in SHP2 are known to cause both Noonan syndrome (NS) and LEOPARD syndrome (LS), two clinically similar autosomal dominant developmental disorders. NS-associated SHP2 mutants display elevated phosphatase activity, while LS-associated SHP2 mutants exhibit reduced catalytic activity. A conundrum in how clinically similar diseases result from mutations to SHP2 that have opposite effects on this enzyme's catalytic functionality exists. Here we report a comprehensive investigation of the kinetic, structural, dynamic, and biochemical signaling properties of the wild type as well as all reported LS-associated SHP2 mutants. The results reveal that LS-causing mutations not only affect SHP2 phosphatase activity but also induce a weakening of the intramolecular interaction between the N-SH2 and PTP domains, leading to mutants that are more readily activated by competing pTyr ligands. Our data also indicate that the residual phosphatase activity associated with the LS SHP2 mutant is required for enhanced ERK1/2 activation. Consequently, catalytically impaired SHP2 mutants could display gain-of-function properties because of their ability to localize to the vicinity of substrates for longer periods of time, thereby affording the opportunity for prolonged substrate turnover and sustained RAS-ERK1/2 activation.


The effect of rotating platform TKA on strain distribution and torque transmission on the proximal tibia.
Limited experimental data exist comparing the mechanical response of the tibial cortex between fixed and rotating platform (RP) total knee arthroplasty (TKA), particularly in the revision setting. We asked if RP-TKA significantly affects tibiofemoral torque and cortical stain response in both the primary and revision settings. Fixed and RP tibial trays were implanted into analogue tibias and biomechanically tested under axial and torsional loading. Torque and strain response were analyzed using digital image correlation. Fixed bearing designs exhibited 13.8 times greater torque (P<0.01), and 69% (P<0.01) higher cortical strain than RP designs. Strain response was similar in the primary and revision cohorts. The decrease in torque transfer could act as a safeguard to reduce stress, micromotion and torsional fatigue in scenario of poor bone stock.


Similar electrophysiological changes in axotomized and neighboring intact dorsal root ganglion neurons.
We investigated electrophysiological changes in chronically axotomized and neighboring intact dorsal root ganglion (DRG) neurons in rats after either a peripheral axotomy consisting of an L5 spinal nerve ligation (SNL) or a central axotomy produced by an L5 partial rhizotomy (PR). SNL produced lasting hyperalgesia to punctate indentation and tactile allodynia to innocuous stroking of the foot ipsilateral to the injury. PR produced ipsilateral hyperalgesia without allodynia with recovery by day 10. Intracellular recordings were obtained in vivo from the cell bodies (somata) of axotomized and intact DRG neurons, some with functionally identified peripheral receptive fields. PR produced only minor electrophysiological changes in both axotomized and intact somata in L5 DRG. In contrast, extensive changes were observed after SNL in large- and medium-sized, but not small-sized, somata of intact (L4) as well as axotomized (L5) DRG neurons. These changes included (in relation to sham values) higher input resistance, lower current and voltage thresholds, and action potentials with longer durations and slower rising and falling rates. The incidence of spontaneous activity, recorded extracellularly from dorsal root fibers in vitro, was significantly higher (in relation to sham) after SNL but not after PR, and occurred in myelinated but not unmyelinated fibers from both L4 (9.1%) and L5 (16.7%) DRGs. We hypothesize that the changes in the electrophysiological properties of axotomized and intact DRG neurons after SNL are produced by a mechanism associated with Wallerian degeneration and that the hyperexcitability of intact neurons may contribute to SNL-induced hyperalgesia and allodynia.


Diffusion maps and coarse-graining: A unified framework for dimensionality reduction, graph partitioning, and data set parameterization.
We provide evidence that nonlinear dimensionality reduction, clustering, and data set parameterization can be solved within one and the same framework. The main idea is to define a system of coordinates with an explicit metric that reflects the connectivity of a given data set and that is robust to noise. Our construction, which is based on a Markov random walk on the data, offers a general scheme of simultaneously reorganizing and subsampling graphs and arbitrarily shaped data sets in high dimensions using intrinsic geometry. We show that clustering in embedding spaces is equivalent to compressing operators. The objective of data partitioning and clustering is to coarse-grain the random walk on the data while at the same time preserving a diffusion operator for the intrinsic geometry or connectivity of the data set up to some accuracy. We show that the quantization distortion in diffusion space bounds the error of compression of the operator, thus giving a rigorous justification for k-means clustering in diffusion space and a precise measure of the performance of general clustering algorithms.


Structure of two related rat pancreatic trypsin genes.
A family of approximately 10 trypsin genes was detected in a rat genomic library by hybridization and in vivo recombination techniques using cloned rat pancreatic trypsin I and II cDNAs as probes. Two separate clones containing the entire trypsin I gene and most of the trypsin II gene were sequenced. Four introns split the trypsin I coding sequence. The positions of the first three introns of the trypsin II gene are identical with those in the trypsin I gene (the fourth intron was not present in the trypsin II clone). The coding regions of the two genes are 88% homologous; the 5'-noncoding regions are 92% homologous, whereas the 3'-noncoding regions share 66% identity. In contrast, the proximal 5'-flanking regions from -1 to -500 which may contain the elements controlling gene expression are less than 30% conserved overall, but segments of approximately 70% homology can be discerned in this region. Some of these sequences are homologous to sequences found in the chymotrypsin and elastase genes. More distal upstream sequences (-500 to -2500) and the intervening sequences show no evident sequence homology (less than 20%). Unique sequences containing homopolymeric purine/pyrimidine repeats are found 2.5 kilobases upstream from the start of transcription of the trypsin I gene and within the second and third introns of the trypsin II gene. The nucleotide homologies as well as the similarities of intron positions of the two trypsin genes to those of other serine protease genes clearly support an evolutionary relationship between members of this gene family.


Dopamine receptor interacting proteins: targeting neuronal calcium sensor-1/D2 dopamine receptor interaction for antipsychotic drug development.
D2 dopamine receptors (D2Rs) represent an important class of receptors in the pharmacological development of novel therapeutic drugs for the treatment of schizophrenia. Recent research into D2R signaling suggests that receptor properties are dependent on interaction with a cohort of dopamine receptor interacting proteins (DRIPs) within a macromolecular structure termed the signalplex. One component of this signalplex is neuronal calcium sensor 1 (NCS-1) a protein found to regulate the phosphorylation, trafficking, and signaling profile of the D2R in neurons. It has also been found that NCS-1 can contribute to the pathology of schizophrenia and may play a role in the efficacy of antipsychotic drug medication in the brain. In this review we discuss how the selective targeting of a DRIP, such as NCS-1, can be utilized as a novel strategy of drug design for the creation of new therapeutics for a disease such as schizophrenia. Using a fluorescence polarization assay we explore how the ability to detect changes in D2R/NCS-1 interaction can be exploited as an effective screening tool in the isolation and development of lead compounds for antipsychotic drug development. This line of work explores a novel direction in targeting D2Rs via their signalplex components and supports the notion that receptor interacting proteins represent an emerging new class of molecular targets for pharmacological drug development.


Quorum-sensing effects in the antagonistic rhizosphere bacterium Serratia plymuthica HRO-C48.
The rhizosphere-associated bacterium Serratia plymuthica HRO-C48 is not only able to suppress symptoms caused by soil-borne pathogens but is also able to stimulate growth of plants. Detailed knowledge about the underlying mechanisms and regulation are crucial for the application in biocontrol strategies. To analyse the influence of N-acyl homoserine lactone (AHL)-mediated communication on the biocontrol activity, the AHL-degrading lactonase AiiA was heterologously expressed in the strain, resulting in abolished AHL production. The comparative analysis of the wild type and AHL negative mutants led to the identification of new AHL-regulated phenotypes. In the pathosystem Verticillium dahliae-oilseed rape, the essential role of AHL-mediated signaling for disease suppression was demonstrated. In vitro, the regulatory function of AHLs in the synthesis of the plant growth hormone indole-3-acetic acid is shown for the first time. Additionally, swimming motility was found to be negatively AHL regulated. In contrast, production of extracellular hydrolytic enzymes is shown to be positively AHL-regulated. HRO-C48 emits a broad spectrum of volatile organic compounds that are involved in antifungal activity and, interestingly, whose relative abundances are influenced by quorum sensing (QS). This study shows that QS is crucial for biocontrol activity of S. plymuthica and discusses the impact for the application of the strain as a biocontrol agent.


Antigen-driven selection of antibodies against SSA, SSB and the centromere 'complex', including a novel antigen, MIS12 complex, in human salivary glands.
Recent evidences have revealed that anti-SSA/SSB antibodies, the major autoantibodies in Sjögren's syndrome (SS), are produced in salivary glands. This study aims to clarify overall of autoantibody production at lesion site, including anti-centromere antibody (ACA)-positive SS. Antibodies of antibody-secreting cells in human salivary glands were produced as recombinant antibodies. The reactivity of these antibodies and their revertants were investigated by ELISA and newly developed antigen-binding beads assay, which can detect conformational epitopes. The target of uncharacterised antibodies was identified by immunoprecipitation and mass spectrometry. Autoantibody-secreting cells in salivary gland tissue were identified by immunohistochemistry using green fluorescent protein-autoantigen fusion proteins. A total of 256 lesion antibodies were generated, and 69 autoantibodies including 24 ACAs were identified among them. Beads assay could detect more autoantibodies than ELISA, suggesting autoantibodies target to antigens with native conformation. After somatic hypermutations were reverted, autoantibodies drastically decreased antigen reactivity. We showed that MIS12 complex, a novel target of ACA, and CENP-C are major targets of ACA produced in salivary glands by examining cloned antibodies and immunohistochemistry, whereas few anti-CENP-B antibodies were detected. The target profiling of serum ACA from 269 patients with SS, systemic sclerosis (SSc), primary biliary cirrhosis (PBC) and healthy controls revealed that ACA-positive patients have antibodies against various sites of centromere complex regardless of disease. We showed direct evidences of antigen-driven maturation of anti-SSA/SSB antibody and ACA in SS lesion. ACA recognises centromere 'complex' rather than individual protein, and this feature is common among patients with SS, SSc and PBC.


MiR-346 regulates CD4⁺CXCR5⁺ T cells in the pathogenesis of Graves' disease.
Follicular helper T (Tfh) cells are increasingly recognized as participants in various autoimmune diseases, including Graves' disease. Although many transcription factors and cytokines are known to regulate Tfh cells, the role of noncoding RNA in Tfh cells development and function is poorly understood. Twenty-three patients with GD, eleven patients with remitting GD, and twenty-four healthy controls were enrolled in the current study. The interaction of miRNA and target gene was predicted through software analysis and then validated by luciferase assay and Western blot. The levels of miR-346 in circulating CD4(+) T cells and plasma were measured by qRT-PCR. The correlation of miR-346 levels with the percentages of CD4(+)CXCR5(+)T cells and autoantibody levels were also analyzed. Up-regulation of Bcl-6 and down-regulation of miR-346 in GD patients were observed, and miR-346 could inhibit Bcl-6 at both transcriptional and translational levels. Overexpression of miR-346 led to attenuating CD4(+)CXCR5(+) T cells. The abnormal expression of miR-346 restored in GD patients after treatment. A negative correlation between levels of miR-346 and percentages of CD4(+)CXCR5(+) T cells was confirmed in GD patients. Additionally, negative correlations between the levels of miR-346 in circulating CD4(+) T cells and serum concentrations of TR-Ab, TG-Ab, and TPO-Ab were also revealed in GD patients. MiR-346 regulates CD4(+)CXCR5(+) T cells by targeting Bcl-6, a positive regulator of Tfh cells, and might play an important role in the pathogenesis of Graves' disease.


Induction of Mitotic Delay in Pharyngeal and Nasopharyngeal Carcinoma Cells Using an Aqueous Extract of Ajuga bracteosa.
Nasopharyngeal carcinoma (NPC) is a common cancer in Southeast Asia, for which radiotherapy and/or chemotherapy are the primary treatment methods. Many herbs are known to have potential uses in chemotherapy; however, the mechanisms underlying the observed antitumor activity of Ajuga bracteosa (AB) against NPC remain unclear. We explored the antitumor effects of AB, which was shown specifically to induce mitotic delay in pharyngeal (Detroit 562) and nasopharyngeal (Hone-1) cancer cells. Proliferation of cancer cells after exposure to aqueous extract of A. bracteosa (AEAB) was assessed using the MTT assay. DNA content and cell cycle arrest induction were analyzed using flow cytometry. The expression of checkpoint kinase 2 (CHK2), cell division control protein 2 (CDC2), and cyclin B1 was investigated using qRT-PCR and Western blot analysis. Results indicated the inhibition of cancer cell growth following exposure to AEAB. In addition, AEAB induced the accumulation of G2/M-phase cells in cancer cell through the disassociation of CDC2/cyclin B1 complex. Our findings suggested that, in addition to the known effects of AEAB in NPC prevention, it may have antitumor activities against NPC cells. In conclusion, AEAB inhibits the growth of and induces mitotic delay in cancer cells, supporting its use as an anticancer agent.


Cognitive impairment and mortality in older mexican americans.
To examine the extent to which cognitive status and decline in cognitive status predict mortality in older Mexican Americans. Longitudinal cohort. Older Mexican Americans residing in five southwestern states in the United States. Two thousand six hundred twenty-five persons aged 65 and older living in Texas, New Mexico, Colorado, Arizona, and California. The cognitive function of participants was assessed using the Mini-Mental State Examination (MMSE) in 1993-94 and again in 1995-96. Cognitive decline was defined using two sets of criteria: a drop to 17 or less (moderate-severe cognitive impairment) on the MMSE at 2-year follow-up and a decrease of at least four points, a threshold change in scores that was used to predict mortality in this sample. Cox proportional hazards models were conducted to examine the association between the MMSE and increased risk for mortality, after controlling for sociodemographic characteristics, medical conditions, and depressive symptoms. Five-year mortality risk was significantly associated with persons in the moderately-severely cognitively impaired category (hazard ratio (HR) = 2.35, P <.001). Moreover, mild cognitive impairment was also predictive of mortality in older Mexicans (HR = 1.45, P <.001). Two-year declines in cognitive functioning, particularly in persons who had declined to the moderate-severe impairment category (HR = 2.23, P <.001) and those who dropped at least four points on the MMSE (HR = 1.30, P <.001), were predictive of mortality 3 years later, after controlling for important variables. Baseline moderate-severe and mild cognitive status and 2-year decline in cognitive functioning independently predict mortality in older Mexican Americans. Although age and selected medical conditions have been reported as the more salient predictors of mortality, cognitive functioning should be considered part of identifying older persons at high risk for underlying medical conditions and mortality.


The "systolic volume balance" method for the noninvasive estimation of cardiac output based on pressure wave analysis.
Cardiac output (CO) monitoring is essential for the optimal management of critically ill patients. Several mathematical methods have been proposed for CO estimation based on pressure waveform analysis. Most of them depend on invasive recording of blood pressure and require repeated calibrations, and they suffer from decreased accuracy under specific conditions. A new systolic volume balance (SVB) method, including a simpler empirical form (eSVB), was derived from basic physical principles that govern blood flow and, in particular, a volume balance approach for the conservation of mass ejected into and flowed out of the arterial system during systole. The formulas were validated by a one-dimensional model of the systemic arterial tree. Comparisons of CO estimates between the proposed and previous methods were performed in terms of agreement and accuracy using "real" CO values of the model as a reference. Five hundred and seven different hemodynamic cases were simulated by altering cardiac period, arterial compliance, and resistance. CO could be accurately estimated by the SVB method as follows: CO = C × PP(ao)/(T - P(sm) × T(s)/P(m)) and by the eSVB method as follows: CO = k × C × PP(ao)/T, where C is arterial compliance, PP(ao) is aortic pulse pressure, T is cardiac period, P(sm) is mean systolic pressure, T(s) is systolic duration, P(m) is mean pressure, and k is an empirical coefficient. SVB applied on aortic pressure waves did not require calibration or empirical correction for CO estimation. An empirical coefficient was necessary for brachial pressure wave analysis. The difference of SVB-derived CO from model CO (for brachial waves) was 0.042 ± 0.341 l/min, and the limits of agreement were -0.7 to 0.6 l/min, indicating high accuracy. The intraclass correlation coefficient and root mean square error between estimated and "real" CO were 0.861 and 0.041 l/min, respectively, indicating very good accuracy. eSVB also provided accurate estimation of CO. An in vivo validation study of the proposed methods remains to be conducted.


Molecular and Biochemical Analysis of the Estrogenic and Proliferative Properties of Vitamin E Compounds.
Tocols are vitamin E compounds that include tocopherols (TPs) and tocotrienols (TTs). These lipophilic compounds are phenolic antioxidants and are reportedly able to modulate estrogen receptor β (ERβ). We investigated the molecular determinants that control their estrogenicity and effects on the proliferation of breast cancer cells. Docking experiments highlighted the importance of the tocol phenolic groups for their interaction with the ERs. Binding experiments confirmed that they directly interact with both ERα and ERβ with their isoforms showing potencies in the following order: δ-tocols > γ-tocols > α-tocols. We also found that tocols activated the transcription of an estrogen-responsive reporter gene that had been stably transfected into cells expressing either ERα or ERβ. The role of the phenolic group in tocol-ER interaction was further established using δ-tocopherylquinone, the oxidized form of δ-TP, which had no ER affinity and did not induce ER-dependent transcriptional modulation. Tocol activity also required the AF1 transactivation domain of ER. We found that both δ-TP and δ-TT stimulated the expression of endogenous ER-dependent genes. However, whereas δ-TP induced the proliferation of ER-positive breast cancer cells but not ER-negative breast cancer cells, δ-TT inhibited the proliferation of both ER-positive and ER-negative breast cancer cells. These effects of δ-TT were found to act through the down regulation of HMG-CoA reductase (HMGR) activity, establishing that ERs are not involved in this effect. Altogether, these data show that the reduced form of δ-TP has estrogenic properties which are lost when it is oxidized, highlighting the importance of the redox status in its estrogenicity. Moreover, we have shown that δ-TT has antiproliferative effects on breast cancer cells independently of their ER status through the inhibition of HMGR. These data clearly show that TPs can be discriminated from TTs according to their structure.


The aetiology of head injury in admitted patients in Jamaica.
This cross-sectional, descriptive study identified 857 head-injured patients who were admitted to the University Hospital of the West Indies (UHWI) over a four-year period. Their median age (IQR) was 28 (16, 45) years and 629 (73.5%) were males. Median length of hospital stay (IQR) was 2 (1, 6) days. Median ICU stay in the intensive care unit (IQR) was 6 (2, 12) days for the 59 (6.9%) patients admitted there. Most patients (73.3%) were admitted with unintentional injuries resulting from road traffic accidents (48.9%), of which passengers were the most commonly affected, and from falls which occurred in 24.4%. Intentional injuries accounted for 26.7% of those admitted, consisting mainly of assaults with blunt objects in 18.0% (154/857). Penetrating injuries were less common, accounting for 67 (7.8%) injuries. Among these, there were 23 gunshot wounds of the head (2.7%). Head injury in admitted patients is mainly due to road traffic accidents, falls and interpersonal violence. Prevention and interventional strategies including education, law enforcement, physical and social engineering must focus on these aetiologies. Current measures are clearly insufficient and more effective strategies are urgently warranted.


Evolutionary conservation of residues in vertebrate DNA polymerase N conferring low fidelity and bypass activity.
POLN is a nuclear A-family DNA polymerase encoded in vertebrate genomes. POLN has unusual fidelity and DNA lesion bypass properties, including strong strand displacement activity, low fidelity favoring incorporation of T for template G and accurate translesion synthesis past a 5S-thymine glycol (5S-Tg). We searched for conserved features of the polymerase domain that distinguish it from prokaryotic pol I-type DNA polymerases. A Lys residue (679 in human POLN) of particular interest was identified in the conserved 'O-helix' of motif 4 in the fingers sub-domain. The corresponding residue is one of the most important for controlling fidelity of prokaryotic pol I and is a nonpolar Ala or Thr in those enzymes. Kinetic measurements show that K679A or K679T POLN mutant DNA polymerases have full activity on nondamaged templates, but poorly incorporate T opposite template G and do not bypass 5S-Tg efficiently. We also found that a conserved Tyr residue in the same motif not only affects sensitivity to dideoxynucleotides, but also greatly influences enzyme activity, fidelity and bypass. Protein sequence alignment reveals that POLN has three specific insertions in the DNA polymerase domain. The results demonstrate that residues have been strictly retained during evolution that confer unique bypass and fidelity properties on POLN.


The evolution of the cortico-cerebellar complex in primates: anatomical connections predict patterns of correlated evolution.
Investigations into the evolution of the primate brain have tended to neglect the role of connectivity in determining which brain structures have changed in size, focusing instead on changes in the size of the whole brain or of individual brain structures, such as the neocortex, in isolation. We show that the primate cerebellum, neocortex, vestibular nuclei and relays between them exhibit correlated volumetric evolution, even after removing the effects of change in other structures. The patterns of correlated evolution among individual nuclei correspond to their known patterns of connectivity. These results support the idea that the brain evolved by mosaic size change in arrays of functionally connected structures. Furthermore, they suggest that the much discussed expansion of the primate neocortex should be re-evaluated in the light of conjoint cerebellar expansion.


Cancer chemoprevention: Evidence of a nonlinear dose response for the protective effects of resveratrol in humans and mice.
Resveratrol is widely promoted as a potential cancer chemopreventive agent, but a lack of information on the optimal dose prohibits rationally designed trials to assess efficacy. To challenge the assumption that "more is better," we compared the pharmacokinetics and activity of a dietary dose with an intake 200 times higher. The dose-response relationship for concentrations generated and the metabolite profile of [(14)C]-resveratrol in colorectal tissue of cancer patients helped us to define clinically achievable levels. In Apc(Min) mice (a model of colorectal carcinogenesis) that received a high-fat diet, the low resveratrol dose suppressed intestinal adenoma development more potently than did the higher dose. Efficacy correlated with activation of adenosine monophosphate-activated protein kinase (AMPK) and increased expression of the senescence marker p21. Nonlinear dose responses were observed for AMPK and mechanistic target of rapamycin (mTOR) signaling in mouse adenoma cells, culminating in autophagy and senescence. In human colorectal tissues exposed to low dietary concentrations of resveratrol ex vivo, we measured enhanced AMPK phosphorylation and autophagy. The expression of the cytoprotective NAD(P)H dehydrogenase, quinone 1 (NQO1) enzyme was also increased in tissues from cancer patients participating in our [(14)C]-resveratrol trial. These findings warrant a revision of developmental strategies for diet-derived agents designed to achieve cancer chemoprevention.


Respiratory quotient in patients with non-insulin-dependent diabetes mellitus treated with insulin and oral hypoglycemic agents.
The respiratory quotient (RQ) reflects the amount of energy derived from carbohydrate as apposed to fat metabolism. To assess the metabolic state of patients with non-insulin-dependent diabetes mellitus, the RQ was measured five times a day (at 09.00, 11.00, 13. 00, 14.00, and 17.00 h) in 20 healthy subjects and 60 diabetic patients. Diabetic patients treated with insulin or sulfonylurea showed significantly higher RQ values than normal subjects and nontreated diabetic patients. Diabetic patients without treatment showed higher glucose levels, and their RQ values were significantly lower than those of treated patients. There was a significant inverse correlation between RQ and blood glucose levels at 11.00 h (r = -0.361, p < 0.01) in diabetic patients, but no significant relation with HbA1c. Treated diabetic patients with a higher body mass index tended to show a higher RQ than those with a lower one (r = -0.269, p = 0.083). Within 1 year, 7 of 13 patients, who had RQ > 1.0, gained more than 3 kg, while only 5 of the remaining 32 treated diabetic patients gained more than 3 kg (p < 0.05). This demonstrates that diabetic patients with a higher RQ tended to gain weight despite the use of insulin or oral hypoglycemia agents. The RQ increased by infusing both insulin and glucose in normal subjects. These results suggest that a high RQ results from excess insulin and excess food. The RQ is a good predictor of weight gain in diabetic patients treated with either insulin or oral hypoglycemic agents.


Use of intra-osseous access in adults: a systematic review.
Indications for intra-osseous (IO) infusion are increasing in adults requiring administration of fluids and medications during initial resuscitation. However, this route is rarely used nowadays due to a lack of knowledge and training. We reviewed the current evidence for its use in adults requiring resuscitative procedures, the contraindications of the technique, and modalities for catheter implementation and skill acquisition. A PubMed search for all articles published up to December 2015 was performed by using the terms "Intra-osseous" AND "Adult". Additional articles were included by using the "related citations" feature of PubMed or checking references of selected articles. Editorials, comments and case reports were excluded. Abstracts of all the articles that the search yielded were independently screened for eligibility by two authors and included in the analysis after mutual consensus. In total, 84 full-text articles were reviewed and 49 of these were useful for answering the following question "when, how, and for which population should an IO infusion be used in adults" were selected to prepare independent drafts. Once this step had been completed, all authors met, reviewed the drafts together, resolved disagreements by consensus with all the authors, and decided on the final version. IO infusion should be implemented in all critical situations when peripheral venous access is not easily obtainable. Contraindications are few and complications are uncommon, most of the time bound to prolonged use. The IO infusion allows for blood sampling and administration of virtually all types of fluids and medications including vasopressors, with a bioavailability close to the intravenous route. Unfortunately, IO infusion remains underused in adults even though learning the technique is rapid and easy. Indications for IO infusion use in adults requiring urgent parenteral access and having difficult intravenous access are increasing. Physicians working in emergency departments or intensive care units should learn the procedures for catheter insertion and maintenance, the contraindications of the technique, and the possibilities this access offers.


The unexpected discovery of a novel low-oxygen-activated locus for the anoxic persistence of Burkholderia cenocepacia.
Burkholderia cenocepacia is a Gram-negative aerobic bacterium that belongs to a group of opportunistic pathogens displaying diverse environmental and pathogenic lifestyles. B. cenocepacia is known for its ability to cause lung infections in people with cystic fibrosis and it possesses a large 8 Mb multireplicon genome encoding a wide array of pathogenicity and fitness genes. Transcriptomic profiling across nine growth conditions was performed to identify the global gene expression changes made when B. cenocepacia changes niches from an environmental lifestyle to infection. In comparison to exponential growth, the results demonstrated that B. cenocepacia changes expression of over one-quarter of its genome during conditions of growth arrest, stationary phase and surprisingly, under reduced oxygen concentrations (6% instead of 20.9% normal atmospheric conditions). Multiple virulence factors are upregulated during these growth arrest conditions. A unique discovery from the comparative expression analysis was the identification of a distinct, co-regulated 50-gene cluster that was significantly upregulated during growth under low oxygen conditions. This gene cluster was designated the low-oxygen-activated (lxa) locus and encodes six universal stress proteins and proteins predicted to be involved in metabolism, transport, electron transfer and regulation. Deletion of the lxa locus resulted in B. cenocepacia mutants with aerobic growth deficiencies in minimal medium and compromised viability after prolonged incubation in the absence of oxygen. In summary, transcriptomic profiling of B. cenocepacia revealed an unexpected ability of aerobic Burkholderia to persist in the absence of oxygen and identified the novel lxa locus as key determinant of this important ecophysiological trait.


Hearing Impairment and Cognitive Energy: The Framework for Understanding Effortful Listening (FUEL).
The Fifth Eriksholm Workshop on "Hearing Impairment and Cognitive Energy" was convened to develop a consensus among interdisciplinary experts about what is known on the topic, gaps in knowledge, the use of terminology, priorities for future research, and implications for practice. The general term cognitive energy was chosen to facilitate the broadest possible discussion of the topic. It goes back to who described the effects of attention on perception; he used the term psychic energy for the notion that limited mental resources can be flexibly allocated among perceptual and mental activities. The workshop focused on three main areas: (1) theories, models, concepts, definitions, and frameworks; (2) methods and measures; and (3) knowledge translation. We defined effort as the deliberate allocation of mental resources to overcome obstacles in goal pursuit when carrying out a task, with listening effort applying more specifically when tasks involve listening. We adapted Kahneman's seminal (1973) Capacity Model of Attention to listening and proposed a heuristically useful Framework for Understanding Effortful Listening (FUEL). Our FUEL incorporates the well-known relationship between cognitive demand and the supply of cognitive capacity that is the foundation of cognitive theories of attention. Our FUEL also incorporates a motivation dimension based on complementary theories of motivational intensity, adaptive gain control, and optimal performance, fatigue, and pleasure. Using a three-dimensional illustration, we highlight how listening effort depends not only on hearing difficulties and task demands but also on the listener's motivation to expend mental effort in the challenging situations of everyday life.


Biofield therapies and cancer-related symptoms: a review.
Patients with cancer can experience several treatment-related symptoms, and conventional care focuses primarily on cure and survival without a holistic approach to disease. Subsequently, an increasing number of patients are accustomed to complementary modalities to improve well-being. Biofield therapies (BTs) are complementary and alternative medicine (CAM) modalities based on the philosophy that humans have an energetic dimension. Physical and psychological symptoms may cause imbalance, and BTs are believed to balance disturbance in the energy field. This article provides a study review of the main BTs (i.e., therapeutic touch, healing touch, and Reiki) in the treatment of cancer-related symptoms. Although BTs are among the most ancient healing practices, data on their effectiveness are poor and additional multicenter research with larger samples are necessary. BTs may eventually become an autonomous field of nursing activity and allow professionals to build a relationship with the patient, thereby improving motivation. The idea that this method can be self-managed and may effectively reduce pain for patients with cancer can improve satisfaction challenges experienced by the current healthcare system.


Passive ventricular constraint prevents transmural shear strain progression in left ventricle remodeling.
Passive ventricular constraint provides external cardiac support to reduce left ventricular (LV) wall stress and myocardial stretch, which are primary determinants of LV remodeling. Altered wall strain results in cytokine and reactive oxygen species production, which, in turn, stimulates apoptosis and extracellular matrix disruption and could be an important trigger for adverse global LV dilatation and remodeling. The effects of the Acorn cardiac support device (CSD) on regional transmural LV wall strains, however, remain unknown. Thirty-three sheep had transmural radiopaque beadsets surgically inserted into the anterior basal and lateral equatorial LV walls, with additional markers silhouetting the left ventricle. Eight animals had CSD implanted (myocardial infarction [MI]+CSD). One week thereafter, the MI+CSD group and 10 animals without CSD (MI) underwent posterior LV infarction by snaring obtuse marginal coronary arteries. Fifteen animals (Sham) had no infarction or CSD. 4D marker dynamics were measured with biplane videofluoroscopy 1 and 8 weeks postoperatively. LV volumes, sphericity index, and transmural circumferential, longitudinal, and radial systolic strains were analyzed. Compared with Sham, infarction (MI) dilated the heart, reduced sphericity index (LV length/width), and increased longitudinal-radial shear strains in the inner half of both the anterior and lateral LV walls. CSD prevented this shear strain perturbation, minimized LV end diastolic volume increase, and augmented the LV sphericity index. Prophylactic CSD prevented infarct-induced shear strain progression not only in myocardium adjacent to, but also remote from, the infarct. CSD also prevented LV dilatation and sphericalization. By attenuating shear strain abnormalities, CSD could prevent the heart from entering into a positive feedback loop of further LV dilatation and exaggeration of LV wall stress and may reduce biochemical triggers portending adverse LV remodeling.


Interaction between cholinergic and catecholaminergic agents in a spontaneous alternation task.
Spontaneous alternation was examined in a free running Y-maze task after various pharmacological manipulations. Whereas scopolamine reduced alternation to chance levels, d-amphetamine in some doses resulted in alternation significantly below chance (perservation). Physostigmine treatment increased levels of alternation whereas reserpine was without effect. Concurrent administration of drugs revealed that reserpine effectively reversed the effects of scopolamine, while the perseveration induced by d-amphetamine was antagonized by physostigmine. When animals were pre-exposed to the Y-maze the effects of d-amphetamine were enhanced, but effects of scopolamine were not modified. Finally, scopolamine treatment augmented the perseverative effects of d-amphetamine. It was suggested that cholinergic agents modify alternation by effects on habituation. On the other hand d-amphetamine produces genuine perseveration without effects on habituation per se. Alternation performance and perseveration were suggested to be mediated by the interaction between the distinct behavioral effects of cholinergic and catecholaminergic activity.


Neurological, Psychiatric, and Biochemical Aspects of Thiamine Deficiency in Children and Adults.
Thiamine (vitamin B1) is an essential nutrient that serves as a cofactor for a number of enzymes, mostly with mitochondrial localization. Some thiamine-dependent enzymes are involved in energy metabolism and biosynthesis of nucleic acids whereas others are part of the antioxidant machinery. The brain is highly vulnerable to thiamine deficiency due to its heavy reliance on mitochondrial ATP production. This is more evident during rapid growth (i.e., perinatal periods and children) in which thiamine deficiency is commonly associated with either malnutrition or genetic defects. Thiamine deficiency contributes to a number of conditions spanning from mild neurological and psychiatric symptoms (confusion, reduced memory, and sleep disturbances) to severe encephalopathy, ataxia, congestive heart failure, muscle atrophy, and even death. This review discusses the current knowledge on thiamine deficiency and associated morbidity of neurological and psychiatric disorders, with special emphasis on the pediatric population, as well as the putative beneficial effect of thiamine supplementation in autism spectrum disorder (ASD) and other neurological conditions.


[Adenomatoid tumour of epididymis: the most common tumour of the paratesticular structures].
Paratesticular tumours are rare but generally benign neoplasms, usually treated by local excission. Adenomatoid tumours of epididymis are the most common of these tumours. Analyze paratesticular tumours treated in our center and describe dyagnosis and treatment of adenomatoid tumours of epididymis. We retrospectively review 8 patients with paratesticular tumours treated from July 1997 to July 2007. We analyze clinical presentation, dyagnostic suspicion given by image technique, treatment followed and final dyagnosis. Patients median age was 44.1 years (22-69), presenting most of them subacute scrotal tumour with median size by ultrasound of 2.8 cm (1.5-7). All of them were locally extirpated except one with suspicion of a malignant polyorchidism and another one with an apparently intratesticular lesion of great size. Just in two cases peroperatory biopsy was performed. Dyagnosis was in 4 cases adenomatoid tumour of epididymis, in two cases fibrous pseudotumour of epididymis, in one case leiomyoma of epididymis and in one case angiolipoma of spermatic cord. Just in one case diagnosed of adenomatoid tumour of epididymis ultrasound confirmed solid tumour suggesting the final dyagnosis. Adenomatoid tumors of epididymis are rare tumours which may present at any age. Ultrasound may help in dyagnosis, but its capacity to distinguish this lesions is low. Benignity of adenomatoid tumour of epididymis as well as most of the other paratesticular tumours should make local excission the treatment of choice and, when any doubt existed, peroperatory biopsy should be performed.


Frailty Is Associated With Neutrophil Dysfunction Which Is Correctable With Phosphoinositol-3-Kinase Inhibitors.
Neutrophil dysfunction has been described with age, appears exaggerated in infection, with altered phosphoinositol signaling a potential mechanism. However, functional aging is heterogeneous. Frailty is a negative health status and is more common in older adults. We hypothesized that neutrophil migration may be compromised in frailty, associated with the degree of frailty experienced by the older person. We compared measures of frailty, neutrophil function, and systemic inflammation in 40 young and 77 older community-dwelling adults in the United Kingdom. Systemic neutrophils exhibited an age-associated reduction in the accuracy of migration (chemotaxis) which was further blunted with frailty. The degree of migratory inaccuracy correlated with physical (adjusted hand grip strength) and cognitive (Stroop test) markers of frailty. Regression analysis demonstrated that age, Charlson comorbidity index, and frailty index were able to predict neutrophil chemotaxis. Reduced chemotaxis of neutrophils from frail adults could be reversed using selective PI3K inhibitors. Exposure of neutrophils from young adults to plasma from chronically inflamed frail older adults could not recapitulate the migratory deficit in vitro, and there were no relationships with systemic inflammation and neutrophil dysfunction. Frailty exaggerated the neutrophil deficits seen with advanced age but aspects of the frailty-associated deficit in neutrophil function are rescuable and thus potentially form a therapeutic target to improve outcomes from infection in older adults.


Balloon test occlusion of the internal carotid artery: change in stump pressure over 15 minutes and its correlation with xenon CT cerebral blood flow.
To determine whether stump pressure changes significantly over time during temporary internal carotid artery (ICA) occlusion via an endovascular balloon, and to examine correlations between stump pressure changes and cerebral blood flow (CBF) as measured by xenon CT. Seventy candidates for ICA sacrifice were evaluated preoperatively with a test occlusion of the cervical ICA using an endovascular balloon. Measurements of ICA stump pressure above the occlusive balloon through a distal lumen in the balloon catheter were made throughout the 15-minute test. During occlusion, CBF was measured with stable xenon CT techniques. Patients were then categorized according to CBF results. The stump pressure changed significantly (P < .001) during the occlusive period. On average, stump pressure increased 7.7 mmHg (12%). Patients determined by xenon CT to have an asymmetric decrease in CBF with balloon occlusion did not display the upward trend in stump pressures and were significantly different (P = .013) in that respect from patients who demonstrated either no CBF change or only minimal bilaterally symmetric decreases in CBF. Large overlaps among the groups prohibit the use of stump pressure changes as an accurate predictor of CBF or of stroke risk in an individual patient. However, these data suggest an autoregulatory mechanism in the cerebral circulation that evolves over a period of minutes in contrast to the traditional concept of cerebral autoregulation which is thought to occur in a matter of seconds.


Strategies for Teachers to Promote Physical Activity in Early Childhood Education Settings-A Scoping Review.
Recent evidence has emphasized the importance of the early childhood years for developing lifelong physical activity patterns. As such, evidence-informed programs that create opportunities for young children to engage in physical activity are needed and education settings present an important context. This review aimed to identify strategies that are implemented by teachers to promote physical activity in early childhood education and care settings. This is a scoping review that followed the framework proposed by the Joanna Briggs Institute. Searches were conducted using the databases of PubMed, SCOPUS, PsycINFO, SPORT Discus, ERIC and Web of Science for publications up to September 2020. From a total of 8974 articles, 19 were deemed eligible. Ten types of strategies, performed by teachers with the intention to improve physical activity-related primary outcomes, were identified. Physical activity promotion by teachers in early childhood settings is recommended to take a multi-strategy approach, in conjunction with professional development training opportunities and continuous follow-up support for teachers. Future work is warranted to fill the evidence gap in other regions (e.g., Asia, Africa and South America) and strengthen the evidence base to establish best practice standards.


Pharmacokinetic modeling of [11C]flumazenil kinetics in the rat brain.
Preferred models for the pharmacokinetic analysis of [11C]flumazenil human studies have been previously established. However, direct translation of these models and settings to animal studies might be sub-optimal. Therefore, this study evaluates pharmacokinetic models for the quantification of [11C]flumazenil binding in the rat brain. Dynamic (60 min) [11C]flumazenil brain PET scans were performed in two groups of male Wistar rats (tracer dose (TD), n = 10 and pre-saturated (PS), n = 2). Time-activity curves from five regions were analyzed, including the pons (pseudo-reference region). Distribution volume (VT) was calculated using one- and two-tissue compartment models (1TCM and 2TCM) and spectral analysis (SA). Binding potential (BPND) was determined from full and simplified reference tissue models with one or two compartments for the reference tissue (FRTM, SRTM, and SRTM-2C). Model preference was determined by Akaike information criterion (AIC), while parameter agreement was assessed by linear regression, repeated measurements ANOVA and Bland-Altman plots. 1TCM and 2TCM fits of regions with high specific binding showed similar AIC, a preference for the 1TCM, and good VT agreement (0.1% difference). In contrast, the 2TCM was markedly preferred and necessary for fitting low specific-binding regions, where a worse VT agreement (17.6% difference) and significant VT differences between the models (p < 0.005) were seen. The PS group displayed results similar to those of low specific-binding regions. All reference models (FRTM, SRTM, and SRTM-2C) resulted in at least 13% underestimation of BPND. Although the 1TCM was sufficient for the quantification of high specific-binding regions, the 2TCM was found to be the most adequate for the quantification of [11C]flumazenil in the rat brain based on (1) higher fit quality, (2) lower AIC values, and (3) ability to provide reliable fits for all regions. Reference models resulted in negatively biased BPND and were affected by specific binding in the pons of the rat.


Biological and biochemical characterization of a cloned Leu-3- cell surviving infection with the acquired immune deficiency syndrome retrovirus.
Leu-3- cells that survive infection with the acquired immune deficiency syndrome (AIDS) retrovirus can be induced with IUdR to express infectious virus. A cellular clone (8E5), isolated by limiting dilution of a mass culture of survivor cells, was found to contain a single, integrated provirus that was constitutively expressed. Although IUdR treatment of 8E5 cells failed to induce infectious virus, cocultivation with Leu-3+ cells generated the characteristic syncytia associated with acute AIDS retrovirus infection. The single integrated copy of proviral DNA directs the synthesis of all major viral structural proteins except p64, as monitored by immunoblotting. The relationship of the 8E5 clone to viral latency and persistence is discussed.


ACCP Clinical Pharmacist Competencies.
The purpose of the American College of Clinical Pharmacy (ACCP) is to advance human health by extending the frontiers of clinical pharmacy. Consistent with this mission and its core values, ACCP is committed to ensuring that clinical pharmacists possess the knowledge, skills, attitudes, and behaviors necessary to deliver comprehensive medication management (CMM) in team-based, direct patient care environments. These components form the basis for the core competencies of a clinical pharmacist and reflect the competencies of other direct patient care providers. This paper is an update to a previous ACCP document and includes the expectation that clinical pharmacists be competent in six essential domains: direct patient care, pharmacotherapy knowledge, systems-based care and population health, communication, professionalism, and continuing professional development. Although these domains align with the competencies of physician providers, they are specifically designed to better reflect the clinical pharmacy expertise required to provide CMM in patient-centered, team-based settings. Clinical pharmacists must be prepared to complete the education and training needed to achieve these competencies and must commit to ongoing efforts to maintain competence through ongoing professional development. Collaboration among stakeholders will be needed to ensure that these competencies guide clinical pharmacists' professional development and evaluation by educational institutions, postgraduate training programs, professional societies, and employers.


Prostate specific antigen (PSA) kinetic as a prognostic factor in metastatic prostate cancer receiving androgen deprivation therapy: systematic review and meta-analysis.
Aim: Metastatic prostate cancer (mPCa) has a poor outcome with median survival of two to five years. The use of androgen deprivation therapy (ADT) is a gold standard in management of this stage. Aim of this study is to analyze the prognostic value of PSA kinetics of patient treated with hormonal therapy related to survival from several published studies Method: Systematic review and meta-analysis was performed using literature searching in the electronic databases of MEDLINE, Science Direct, and Cochrane Library. Inclusion criteria were mPCa receiving ADT, a study analyzing Progression Free Survival (PFS), Overall Survival (OS), or Cancer Specific Survival (CSS) and prognostic factor of survival related to PSA kinetics (initial PSA, PSA nadir, and time to achieve nadir (TTN)). The exclusion criteria were metastatic castration resistant of prostate cancer (mCRPC) and non-metastatic disease. Generic inverse variance method was used to combine hazard ratio (HR) within the studies. Meta-analysis was performed using Review Manager 5.2 and a p-value <0.05 was considered statistically significant. Results: We found 873 citations throughout database searching with 17 studies were consistent with inclusion criteria. However, just 10 studies were analyzed in the quantitative analysis. Most of the studies had a good methodological quality based on Ottawa Scale. No significant association between initial PSA and PFS. In addition, there was no association between initial PSA and CSS/ OS. We found association of reduced PFS (HR 2.22; 95% CI 1.82 to 2.70) and OS/ CSS (HR 3.31; 95% CI 2.01-5.43) of patient with high PSA nadir. Shorter TTN was correlated with poor result of survival either PFS (HR 2.41; 95% CI 1.19 - 4.86) or CSS/ OS (HR 1.80; 95%CI 1.42 - 2.30) Conclusion: Initial PSA before starting ADT do not associated with survival in mPCa. There is association of PSA nadir and TTN with survival.


Adhesion properties in systems of laminated pigmented polymers, carbon-graphite fiber composite framework and titanium surfaces in implant suprastructures.
For long-term stability the adhering interfaces of an implant-retained supraconstruction of titanium/carbon-graphite fiber-reinforced (CGFR) polymer/opaquer layer/denture base polymer/denture teeth must function as a unity. The aim was to evaluate adhesion of CGFR polymer to a titanium surface or CGFR polymer to two different opaquer layers/with two denture base polymers. Titanium plates were surface-treated and silanized and combined with a bolt of CGFR polymer or denture base polymer (Probase Hot). Heat-polymerized plates of CGFR polymer (47 wt% fiber) based on poly(methyl methacrylate) and a copolymer matrix were treated with an opaquer (Sinfony or Ropak) before a denture base polymer bolt was attached (Probase Hot or Lucitone 199). All specimens were heat-polymerized, water saturated (200 days) and thermally cycled (5000 cycles, 5/55 degrees C) before shear bond testing. Silicatized titanium surfaces gave higher bond strength to CGFR polymer (16.2+/-2.34 and 18.6+/-1.32) MPa and cohesive fracture than a sandblasted surface (5.9+/-2.11) MPa where the fracture was adhesive. The opaquer Sinfony gave higher adhesion values and mainly cohesive fractures than the opaquer Ropak. Different surface treatments (roughened or polished) of the CGFR polymer had no effect on bond strength. The fracture surfaces of silicatized titanium/CGFR polymer/opaquer layer (Sinfony)/denture base polymers were mainly cohesive. A combination of these materials in an implant-retained supraconstruction is promising for in vivo evaluation.


Determination of plasma volume by 113min-labeled transferrin.
The binding property of ionic indium (In3+) with plasma transferrin was utilized for determination of plasma volumes (PV) of whole body and individual organs in small animals. Plasma transferrin from a donor rat was labeled with 15-17 muCi 113mIn/ml plasma and injected into the tested rats. PV were determined either by extrapolation to the dilution at time zero (for whole animals) or by calculation of the ratio, organ radiation: radiation of a plasma unit volume (for organs). The reliability of the method for determination of whole-body PV was ascertained by comparing the results obtained with those obtained simultaneously by the Evans blue dilution method. Whole body PV values obtained by the two methods were similar, with a correlation coefficient (r) of 0.997. The short half life of 113mIn enables it to be used with other nuclides which have similar or different energies in the same sample; indium radiation was counted first and after it had disappeared the activity of the other nuclide could be measured.


G protein-coupled receptor 35: an emerging target in inflammatory and cardiovascular disease.
G protein-coupled receptor 35 (GPR35) is an orphan receptor, discovered in 1998, that has garnered interest as a potential therapeutic target through its association with a range of diseases. However, a lack of pharmacological tools and the absence of convincingly defined endogenous ligands have hampered the understanding of function necessary to exploit it therapeutically. Although several endogenous molecules can activate GPR35 none has yet been confirmed as the key endogenous ligand due to reasons that include lack of biological specificity, non-physiologically relevant potency and species ortholog selectivity. Recent advances have identified several highly potent synthetic agonists and antagonists, as well as agonists with equivalent potency at rodent and human orthologs, which will be useful as tool compounds. Homology modeling and mutagenesis studies have provided insight into the mode of ligand binding and possible reasons for the species selectivity of some ligands. Advances have also been made in determining the role of the receptor in disease. In the past, genome-wide association studies have associated GPR35 with diseases such as inflammatory bowel disease, type 2 diabetes, and coronary artery disease. More recent functional studies have implicated it in processes as diverse as heart failure and hypoxia, inflammation, pain transduction and synaptic transmission. In this review, we summarize the progress made in understanding the molecular pharmacology, downstream signaling and physiological function of GPR35, and discuss its emerging potential applications as a therapeutic target.


Clear cell adenocarcinoma of the vagina and cervix. An update of the central Netherlands registry showing twin age incidence peaks.
The objective of this study was to update the registry of women in the Netherlands with clear cell adenocarcinoma (CCAC) of the cervix or vagina with or without intrauterine exposure to diethylstilbestrol (DES). From a nationwide search in PALGA, the automated pathology registry in the Netherlands, data were gathered on women with CCAC born after 1947. Information obtained from the clinical files of the patients included reported exposure to DES, patterns of complaints previous to diagnosis, the current status of the patients, and the results of cytopathologic examinations previous to histopathologic diagnosis. After review of the histopathologic slides, the specific pathologic characteristics of CCAC were determined. The age distribution of women born after 1947 was compared with that of women born before 1947. Information about possible exposure to DES during pregnancy was available for 73 of 88 women with CCAC born after 1947. Exposure to DES was reported for 47 (64%) of these women. The DES medication was most often reported as having started before the 18th week of pregnancy. Cytopathologic examination was informative in 81% of the cases of CCAC of the cervix, but only in 41% of the cases of CCAC of the vagina. Most patients had Stage I or II tumors at diagnosis. Tumor Stage III and IV and a high grade of nuclear atypia were related to unfavorable outcome. The age distribution of all patients with CCAC showed two distinct peaks; one at young age, (a mean age of 26 years), and one at older age (a mean age of 71 years). This bimodal age distribution still applied when the cases in which DES exposure was reported had been excluded. Despite the fact that DES has not been prescribed to pregnant women in the Netherlands in the last 20 years, CCAC is still relevant in our times. It is important to stay alert and periodically to update and evaluate the data of this registry, including data on women born outside the DES exposure period. The bimodal age distribution in this study of women without intrauterine exposure to DES suggests a carcinogenesis-promoting role of menarche and menopause and/ or the existence of a subpopulation with genetic risk factors or exogenous risk factors other than exposure to DES. Postmenopausal observation of women exposed to DES must be encouraged for clinical reasons and may help facilitate differentiation between these two hypotheses. If these risk factors of CCAC were better documented and their interrelationships better defined, CCAC could become an important model of multistep carcinogenesis in tissues sensitive to sex hormones.


Intracellular routing of wild-type and mutated polymeric immunoglobulin receptor in hippocampal neurons in culture.
Certain epithelial cells synthesize the polymeric immunoglobulin receptor (pIgR) to transport immunoglobulins (Igs) A and M into external secretions. In polarized epithelia, newly synthesized receptor is first delivered to the basolateral plasma membrane and is then, after binding the Ig, transcytosed to the apical plasma membrane, where the receptor-ligand complex is released by proteolytic cleavage. In a previous work (Ikonen et al., 1993), we implied the existence of a dendro-axonal transcytotic pathway for the rabbit pIgR expressed in hippocampal neurons via the Semliki Forest Virus (SFV) expression system. By labeling surface-exposed pIgR in live neuronal cells, we now show (a) internalization of the receptor from the dendritic plasma membrane to the dendritic early endosomes, (b) redistribution of the receptor from the dendritic to the axonal domain, (c) inhibition of this movement by brefeldin A (BFA) and (d) stimulation by the activation of protein kinase C (PKC) via phorbol myristate acetate (PMA). In addition, we show that a mutant form of the receptor lacking the epithelial basolateral sorting signal is directly delivered to the axonal domain of hippocampal neurons. Although this mutant is internalized into early endosomes, no transcytosis to the dendrites could be observed. In epithelial Madin-Darby Canine Kidney (MDCK) cells, the mutant receptor could also be internalized into basolaterally derived early endosomes. These results suggest the existence of a dendro-axonal transcytotic pathway in neuronal cells which shares similarities with the basolateral to apical transcytosis in epithelial cells and constitute the basis for the future analysis of its physiological role.


The sleeping brain's influence on verbal memory: boosting resistance to interference.
Memories evolve. After learning something new, the brain initiates a complex set of post-learning processing that facilitates recall (i.e., consolidation). Evidence points to sleep as one of the determinants of that change. But whenever a behavioral study of episodic memory shows a benefit of sleep, critics assert that sleep only leads to a temporary shelter from the damaging effects of interference that would otherwise accrue during wakefulness. To evaluate the potentially active role of sleep for verbal memory, we compared memory recall after sleep, with and without interference before testing. We demonstrated that recall performance for verbal memory was greater after sleep than after wakefulness. And when using interference testing, that difference was even more pronounced. By introducing interference after sleep, this study confirms an experimental paradigm that demonstrates the active role of sleep in consolidating memory, and unmasks the large magnitude of that benefit.


Numerical modeling of solid-phase microextraction: binding matrix effect on equilibrium time.
Solid-phase microextraction (SPME) is a well-known sampling and sample preparation technique used for a wide variety of analytical applications. As there are various complex processes taking place at the time of extraction that influence the parameters of optimum extraction, a mathematical model and computational simulation describing the SPME process is required for experimentalists to understand and implement the technique without performing multiple costly and time-consuming experiments in the laboratory. In this study, a mechanistic mathematical model for the processes occurring in SPME extraction of analyte(s) from an aqueous sample medium is presented. The proposed mechanistic model was validated with previously reported experimental data from three different sources. Several key factors that affect the extraction kinetics, such as sample agitation, fiber coating thickness, and presence of a binding matrix component, are discussed. More interestingly, for the first time, shorter or longer equilibrium times in the presence of a binding matrix component were explained with the help of an asymptotic analysis. Parameters that contribute to the variation of the equilibrium times are discussed, with the assumption that one binding matrix component is present in a static sample. Numerical simulation results show that the proposed model captures the phenomena occurring in SPME, leading to a clearer understanding of this process. Therefore, the currently presented model can be used to identify optimum experimental parameters without the need to perform a large number of experiments in the laboratory.


Menin, a tumor suppressor, associates with nonmuscle myosin II-A heavy chain.
MEN1 is a likely tumor suppressor gene that encodes a novel protein, menin. Menin is a 610 amino-acid residue protein with as yet unknown function(s). We have used tandem affinity purification and mass spectroscopy to isolate and identify proteins associating with menin from cultured HeLa cell extracts. This strategy has resulted in the isolation and identification of nonmuscle myosin type II-A heavy chain (NMHC II-A) as a menin interacting protein. This interaction was confirmed by glutathione-S-transferase pulldown assays, by coimmunoprecipitation, and by actin selection of myosin. We have further identified the amino-terminal region of menin and the head domain of NMHC II-A to be regions required for this interaction. Moreover menin was seen to colocalize with this myosin isoform in the cleavage furrow of dividing cells by indirect immunofluoresence. These data indicate that menin through binding to NMHC II-A could participate in cell division and in other processes that involve NMHC II-A.


The Etv2-miR-130a Network Regulates Mesodermal Specification.
MicroRNAs (miRNAs) are known to regulate critical developmental stages during embryogenesis. Here, we defined an Etv2-miR-130a cascade that regulates mesodermal specification and determination. Ablation of Dicer in the Etv2-expressing precursors resulted in altered mesodermal lineages and embryonic lethality. We identified miR-130a as a direct target of Etv2 and demonstrated its role in the segregation of bipotent hemato-endothelial progenitors toward the endothelial lineage. Gain-of-function experiments demonstrated that miR-130a promoted the endothelial program at the expense of the cardiac program without impacting the hematopoietic lineages. In contrast, CRISPR/Cas9-mediated knockout of miR-130a demonstrated a reduction of the endothelial program without affecting hematopoiesis. Mechanistically, miR-130a directly suppressed Pdgfra expression and promoted the endothelial program by blocking Pdgfra signaling. Inhibition or activation of Pdgfra signaling phenocopied the miR-130a overexpression and knockout phenotypes, respectively. In summary, we report the function of a miRNA that specifically promotes the divergence of the hemato-endothelial progenitor to the endothelial lineage.


Cost-effectiveness of different strategies for treatment relapsing-remitting multiple sclerosis.
To compare the cost-effectiveness of different disease-modifying therapies' strategies for treatment of relapsing-remitting multiple sclerosis. A Markov model was developed to assess the cost-effectiveness and incremental cost-effectiveness ratios for different strategies of using disease-modifying therapies from a US third-party payer perspective. All costs were converted to 2014 US$. Over 20 years, the total costs per patient were estimated at US$161,136.60 for Strategy 1 (symptom management [SM] alone), US$551,650.66 for Strategy 2 (SM and IFN-β-1a), US$703,463.60 for Strategy 3 (SM and natalizumab) and US$670,985.24 for Strategy 4 (SM and alemtuzumab). The accumulated quality-adjusted life years were 10.49, 10.66, 10.69 and 10.71 for each of the four Strategies 1-4, respectively. The resulting incremental cost-effectiveness ratios were 2,297,141.53 comparing Strategy 2 to Strategy 1, and -1,623,918.00 comparing Strategy 4 to Strategy 3. Strategy 1 was the cost-effective strategy for treatment of relapsing-remitting multiple sclerosis when compared with other strategies.


Novel amplification targets for rapid detection and differentiation of Xylella fastidiosa subspecies fastidiosa and multiplex in plant and insect tissues.
Xylella fastidiosa is an insect-transmitted bacterial plant pathogen which causes a variety of economically important diseases worldwide. Molecular identification of X. fastidiosa is used for quarantine screening, surveillance, and research applications; many of which require subspecies level differentiation of pathogen isolates. This study describes quantitative PCR (qPCR) and isothermal amplification assays which can rapidly identify X. fastidiosa isolates belonging to the fastidiosa and multiplex subspecies. The TaqMan qPCR primers described here are used to differentiate X. fastidiosa strains by subspecies in plant and insect tissue in a single reaction, with the inclusion of a general amplification control probe to identify potential false negative samples. This TaqMan qPCR protocol can identify between 103 and 104 cfu/ml concentrations of X. fastidiosa at the subspecies level in a variety of sample types. Additionally, loop-mediated isothermal amplification (LAMP) targets were designed for faster detection of X. fastidiosa subspecies fastidiosa and multiplex, applicable to a field setting. These assays are effective for strain differentiation in artificially and naturally inoculated plant material, and in field collected insect vectors.


Surgical checklists: a systematic review of impacts and implementation.
Surgical complications represent a significant cause of morbidity and mortality with the rate of major complications after inpatient surgery estimated at 3-17% in industrialised countries. The purpose of this review was to summarise experience with surgical checklist use and efficacy for improving patient safety. A search of four databases (MEDLINE, CINAHL, EMBASE and the Cochrane Database of Controlled Trials) was conducted from 1 January 2000 to 26 October 2012. Articles describing actual use of the WHO checklist, the Surgical Patient Safety System (SURPASS) checklist, a wrong-site surgery checklist or an anaesthesia equipment checklist were eligible for inclusion (this manuscript summarises all but the anaesthesia equipment checklists, which are described in the Agency for Healthcare Research and Quality publication). We included a total of 33 studies. We report a variety of outcomes including avoidance of adverse events, facilitators and barriers to implementation. Checklists have been adopted in a wide variety of settings and represent a promising strategy for improving the culture of patient safety and perioperative care in a wide variety of settings. Surgical checklists were associated with increased detection of potential safety hazards, decreased surgical complications and improved communication among operating staff. Strategies for successful checklist implementation included enlisting institutional leaders as local champions, incorporating staff feedback for checklist adaptation and avoiding redundancies with existing systems for collecting information. Surgical checklists represent a relatively simple and promising strategy for addressing surgical patient safety worldwide. Further studies are needed to evaluate to what degree checklists improve clinical outcomes and whether improvements may be more pronounced in particular settings.


Bioactivity of arid region honey: an in vitro study.
Antioxidant and anti-inflammatory properties of honey have been largely recognized by various studies. Almost all of the potential benefits are associated with polyphenol content. Honey varieties from the arid region are reported to be rich in polyphenols, but data related to its bioactivity in vitro is greatly lacking. This study aimed at establishing the antioxidant and anti-inflammatory properties of arid region honey. Four honey varieties from arid region (H1, H2, H3, and H4) and two popular non-arid region honey (H5 and H6) were tested in vitro in this study. The erythrocyte membrane protection effect of honey varieties were measured by hemolysis assay after exposing erythrocytes to a peroxide generator. The subsequent production of MDA (malondialdehyde) content in erythrocytes was measured. Immunomodulatory effect of the honey varieties was tested in prostate cancer cells PC-3 and PBMC (peripheral blood mononuclear cells) by measuring the IL-6 (interleukin 6) and NO (nitric oxide) levels in cell culture supernatant after incubation with the honey varieties. PC-3 cell viability was assessed after incubation with honey varieties for 24 h. Arid region honey exhibited superior erythrocyte membrane protection effect with H4 measuring 1.3 ± 0.042mMTE/g and H2 measuring 1.122 ± 0.018mMTE/g. MDA levels were significantly reduced by honey samples, especially H4 (20.819 ± 0.63 nmol/mg protein). We observed a significant decrease in cell population in PC-3 after 24 h in culture on treatment with honey. A moderate increase in NO levels was observed in both cultures after 24 h at the same time levels of IL-6 were remarkably reduced by honey varieties. The results demonstrate the antioxidant effect of arid region honey due to its erythrocyte membrane protection effect and subsequent lowering of oxidative damage as evident from lower levels of lipid peroxidation byproduct MDA. Arid region honey varieties were as good as non-arid region types at decreasing cell viability of prostate cancer cells. The moderate increase in NO levels in PC-3 and PBMCs were not significant enough to elicit any pro-inflammatory response. However, IL-6 secretion was remarkably reduced by all honey varieties in a comparable level indicating the potential anti-inflammatory property of arid region honey.


Open tibial shaft fractures: I. Evaluation and initial wound management.
Open fractures of the tibial diaphysis are often associated with severe bone and soft-tissue injury. Contamination of the fracture site and devitalization of the soft-tissue envelope greatly increase the risk of infection, nonunion, and wound complications. Management of open tibial shaft fractures begins with a thorough patient evaluation, including assessment of the bone and soft tissue surrounding the tibial injury. Classification of these injuries according to the system of Gustilo and Anderson at the time of surgical débridement is useful in guiding treatment and predicting outcomes. Administration of antibiotic prophylaxis as soon as possible after injury as well as urgent and thorough débridement, irrigation, and bony stabilization are done to minimize the risk of infection and improve outcomes. The use of antibiotic bead pouches and negative-pressure wound therapy has proved to be efficacious for the acute, temporary management of severe bone and soft-tissue defects.


Pain dominates measurements of elbow function and health status.
Elbow function can be quantified with use of physician-based elbow-rating systems and health status questionnaires. Our hypothesis was that pain has a strong influence on these scores, which overwhelms the influence of objective factors such as motion. One hundred and four patients were evaluated, at a minimum of six months (average, forty-six months) after the latest surgery for an intra-articular fracture of the elbow, with use of three physician-based evaluation instruments (Mayo Elbow Performance Index [MEPI], Broberg and Morrey rating system, and American Shoulder and Elbow Surgeons Elbow Evaluation Instrument [ASES]), an upper-extremity-specific health status questionnaire (Disabilities of the Arm, Shoulder and Hand [DASH]), and a general health status questionnaire (Short Form-36 [SF-36]). Multivariate analysis of variance and regression modeling were used to identify the factors that account for the variability in scores derived with these measures-in other words, which factors have the strongest influence on the final score. Pain alone accounted for 66% of the variability in the MEPI scores, 59% of the variability in the Broberg and Morrey scores, and 57% of the variability in the ASES scores. Models that included other factors accounted for only slightly more variability (73%, 79%, and 79%, respectively), and those that did not include pain accounted for only 22%, 41%, and 41% of the variability. Thirty-six percent of the variability in the DASH scores could be accounted for by pain alone, and 45% could be accounted for by pain and range of motion. Models not including pain accounted for only 17% of the variability in the DASH scores. Pain has a very strong influence on both physician-rated and patient-rated quantitative measures of elbow function. Consequently, these measures may be strongly influenced by the psychosocial aspects of illness that have a strong relationship with pain, and objective measures of elbow function such as mobility may be undervalued. It may be advisable to evaluate pain separately from objective measures of elbow function in physician-based elbow ratings.


Voltage-activated calcium and potassium currents in human pancreatic beta-cells.
1. The whole-cell configuration of the patch clamp technique was used to study inward and delayed outward currents in beta-cells isolated from human pancreatic islets. 2. The delayed outward current activated at about -20 mV and increased linearly with further depolarization. The instantaneous current-voltage (I-V) relation, measured by tail current analysis, reversed at -70 mV. This is close to the K+ equilibrium potential and suggests the outward current is carried primarily by potassium ions. In support of this idea, outward currents were abolished when internal K+ was replaced by the impermeant cation N-methyl-D-glucamine (NMG). 3. The voltage dependence of K+ current activation could be fitted by a sigmoidal function with a mid-point at +1 mV. K+ currents showed voltage-dependent inactivation which was half-maximal at -25 mV. 4. Inward currents were studied after outward currents were suppressed by replacing internal potassium with NMG. In 5 mM [Ca2+]o, the inward current activated between -50 and -40 mV, had a peak amplitude at -10 mV and reversed at potentials positive to +60 mV. The voltage dependence of inward current activation was sigmoidal with half-maximal activation at -10 mV in 5 mM [Ca2+]o and at -22 mV in 5 mM [Ba2+]o. 5. Inward currents were unaffected by tetrodotoxin (TTX), but could be blocked by cadmium ions. Barium was also capable of carrying inward current. This pharmacology is consistent with inward currents flowing through Ca2+ channels. 6. The inactivation of the inward current was dependent on calcium entry. In two-pulse experiments, the voltage dependence of inactivation was U-shaped, and resembled that of the calcium current. Barium currents showed little inactivation. 7. In two-pulse experiments the degree of inward current inactivation during the test pulse was related to the amount of calcium entry during the first pulse. Calcium entering at more positive potentials was less effective at producing inactivation. 8. Calcium and barium currents also showed a slow, voltage-dependent inactivation when the holding potential was changed between -100 and -40 mV. This inactivation developed with a time course of seconds. 9. The Ca2+ and K+ currents described here are similar to those reported for rodent beta-cells and indicate the rodent beta-cell provides a good model for that of man.


Analysis of al-2 mutations in Neurospora.
The orange pigmentation of the fungus Neurospora crassa is due to the accumulation of the xanthophyll neurosporaxanthin and precursor carotenoids. Two key reactions in the synthesis of these pigments, the formation of phytoene from geranylgeranyl pyrophosphate and the introduction of β cycles in desaturated carotenoid products, are catalyzed by two domains of a bifunctional protein, encoded by the gene al-2. We have determined the sequence of nine al-2 mutant alleles and analyzed the carotenoid content in the corresponding strains. One of the mutants is reddish and it is mutated in the cyclase domain of the protein, and the remaining eight mutants are albino and harbor different mutations on the phytoene synthase (PS) domain. Some of the mutations are expected to produce truncated polypeptides. A strain lacking most of the PS domain contained trace amounts of a carotenoid-like pigment, tentatively identified as the squalene desaturation product diapolycopene. In support, trace amounts of this compound were also found in a knock-out mutant for gene al-2, but not in that for gene al-1, coding for the carotene desaturase. The cyclase activity of the AL-2 enzyme from two albino mutants was investigated by heterologous expression in an appropriately engineered E. coli strain. One of the AL-2 enzymes, predictably with only 20% of the PS domain, showed full cyclase activity, suggesting functional independence of both domains. However, the second mutant showed no cyclase activity, indicating that some alterations in the phytoene synthase segment affect the cyclase domain. Expression experiments showed a diminished photoinduction of al-2 transcripts in the al-2 mutants compared to the wild type strain, suggesting a synergic effect between reduced expression and impaired enzymatic activities in the generation of their albino phenotypes.


Comparison of Prehospital Glucose with or without IV Thiamine.
Loading of thiamine prior to glucose administration during hypoglycemia to prevent Wernicke's encephalopathy is routine in the prehospital setting. To date no study has looked at the validity of this therapy. We evaluated a retrospective cohort of 242 patients who received intravenous glucose for hypoglycemia comparing those who received thiamine supplementation versus those who did not. Study endpoints were heart rate, blood pressure, Glasgow Coma Scale (GCS), reentry into the 911 system, and emergency department (ED) discharge rates. There were no significant differences between the thiamine, and without-thiamine groups. All patients were discharged neurologically intact or were alert and oriented when refusing transport to the hospital. None of the 242 patients re-called 911 within the immediate 24-hour period or returned to the ED. To our knowledge this is the first study in the literature which evaluated the use of thiamine with glucose to prevent Wernicke's encephalopathy in the prehospital setting. We found that routine administration of thiamine with glucose did not result in differences in respiratory rate, systolic blood pressure, GCS or ED hospital discharge rates. Until further research is done to validate our results emergency medical services leadership should consider whether the routine use of thiamine in the prehospital setting is appropriate for their system.


Synthesis and characterization of a radioiodinated photoaffinity probe for the alpha 1-adrenergic receptor.
A radioiodinated aryl azide analog, 2-[4-(4-azido-3- iodobenzoyl ) piperazin -1-yl]-4-amino-6, 7- dimethoxyquinazoline [(125I] CP65 ,526), of the highly selective alpha 1-adrenergic antagonist prazosin was synthesized and characterized using rat hepatic plasma membranes. Prior to photolysis, this ligand bound with high affinity (Kd 0.3 nM), stereoselectively and in a saturable manner to sites with an alpha 1-adrenergic specificity. When membranes pretreated with [125I] CP65 ,526 were irradiated with ultraviolet light, the ligand incorporated irreversibly into the receptor-binding sites, also with typical alpha 1-adrenergic specificity. Sodium dodecyl sulphate polyacrylamide gel electrophoresis of such labeled membranes followed by radioautography revealed major bands at Mr = 77,000, 68,000, and 59,000 daltons. Labeling of each of these bands was inhibitable by a variety of adrenergic ligands, stereoselectively and with a specificity typical of the alpha 1-adrenergic receptor. Smaller peptides with molecular weights of 42,000 and 31,000 daltons also displayed prazosin-inhibitable [125] CP65 ,526-binding. However, as the labeling of these protein species was not inhibitable by other adrenergic agonists or antagonists, they are unlikely to represent subunits of the receptor. Further evidence that [125I] CP65 ,526 incorporates covalently upon photolysis was the ability to specifically label immunoglobulin heavy and light chains of an antiserum that recognized both this ligand and the parent compound, prazosin. This new, radioiodinated, high-affinity probe should thus be uniquely valuable for the molecular characterization of the alpha 1-adrenergic receptor.


Components determining the slowness of information processing in parkinson's disease.
Bradyphrenia is a key cognitive feature in Parkinson's disease (PD). There is no consensus on whether information processing speed is impaired or not beyond motor performance. This study aims to explore which perceptual, motor, or cognitive components of information processing are involved in the slowdown affecting cognitive performance. The study included 48 patients with PD (age: 63, 3 ± 8, 18; HY I-III; UPDRS 15,46 ± 7,76) and 53 healthy controls (age: 60,09 ± 12,83). Five reaction time (RT) tasks were administered to all participants. The average RT in each of the tasks and the percentage of correct answers were measured. Patients with PD were in "ON state" at the time of the evaluation. Perceptual, motor, and cognitive components were isolated by means of a series of ANCOVAs. As expected, the motor component was slowed down in patients with PD. Moreover, while patients with PD showed slower RT than controls in all tasks, differences between groups did not exponentially increase with the increasing task complexity. ANCOVA analyses also revealed that the perceptual and sustained alert component resulted to be slowed down, with no differences being found in any of the remaining isolated cognitive components (i.e., response strategy-inhibition, decisional, visual search, or interference control). The results revealed that slowness of information processing in PD was mainly associated with an impaired processing speed of the motor and perceptual-alertness components analyzed. The results may help designing new neurorehabilitation strategies, focusing on the improvement of perceptual and alertness mechanisms.