NK cell natural cytotoxicity and IFN-gamma production are not always coordinately regulated: engagement of DX9 KIR+ NK cells by HLA-B7 variants and target cells.
DX9 mAb-binding killer cell-inhibitory receptors (KIR) recognize HLA-B molecules that express the Bw4 public serologic epitope. We assessed DX9+ NK cell fine specificity recognition of HLA-B7 variants and HLA-B27 alleles by 51Cr release natural cytotoxicity assays and by flow cytometry and enzyme-linked immunospot (ELISPOT) IFN-gamma synthesis and release assays. 721.221 target cell expression of Bw4+ HLA-B27 alleles specifically inhibited DX9+ NK cell natural cytotoxicity and IFN-gamma synthesis and release. A triple substitution of HLA-B7 at residues 80, 82, and 83 known to induce expression of the Bw4 serologic epitope also specifically inhibited DX9+ NK cell natural cytotoxicity and IFN-gamma responses. Single HLA-B7 amino acid substitution variants were recognized in the same decreasing rank order by DX9+ NK cells and Bw4-reactive mAbs: G83R > R82L > N80T = HLA-B7. Natural cytotoxicity inhibition was reversed by the presence of blocking DX9 mAb. Natural cytotoxicity and IFN-gamma production were coordinately regulated by a panel of HLA-B7 variants expressed on 721.221 cells, suggesting that these two effector functions are inhibited by the same KIR-mediated signaling mechanisms. In contrast, some NK cell clones killed 721.221 and K562 target cells equally well but released much more IFN-gamma in response to K562 target cells. Differential regulation of natural cytotoxicity and IFN-gamma release shows that NK cell effector functions respond to distinct signals.


MTH1, an 8-oxo-2'-deoxyguanosine triphosphatase, and MYH, a DNA glycosylase, cooperate to inhibit mutations induced by chronic exposure to oxidative stress of ionising radiation.
Our previous results showed that in addition to the immediate interaction of ionising radiation with DNA (direct and indirect effect), low-dose and chronic low-dose rate of irradiation induce endogenous oxidative stress. During oxidative stress, free radicals react with DNA, nucleoside triphosphates (dNTPs), proteins and lipids, and modify their structures. The MYH and MTH1 genes play important roles in preventing mutations induced by 8-hydroxy-guanine, which is an oxidised product of guanine. In this study, we used short-hairpin RNA to permanently knockdown MYH and MTH1 proteins in human lymphoblastoid TK6 cells. Knockdown and wild-type cells were chronically exposed to low dose rates of γ-radiation (between 1.4 and 30 mGy/h). The cells were also subjected to acute doses delivered at a high-dose rate. Growth rate, extracellular 8-hydroxy-2'-deoxyguanosine, clonogenic cell survival and mutant frequencies were analysed in all cell types. A reduced level of cell growth and survival as well as increased mutant frequencies were observed in cells lacking both MYH and MTH1 proteins as compared to cells lacking only MYH and wild-type cells. To sum up, our results suggest that low-dose rates elevate oxidative stress. MTH1 together with MYH plays an important role in protection against mutations induced by modified dNTPs during chronic oxidative stress. In addition, we found no dose-rate effect at the level of mutations in the wild-type TK6 and MYH-KD cells. Our data interestingly indicate a dose-rate threshold for mutation induction in MTH1/MYH double knockdown cells.


Development of a computerised decision aid for thrombolysis in acute stroke care.
Thrombolytic treatment for acute ischaemic stroke improves prognosis, although there is a risk of bleeding complications leading to early death/severe disability. Benefit from thrombolysis is time dependent and treatment must be administered within 4.5 hours from onset of symptoms, which presents unique challenges for development of tools to support decision making and patient understanding about treatment. Our aim was to develop a decision aid to support patient-specific clinical decision-making about thrombolysis for acute ischaemic stroke, and clinical communication of personalised information on benefits/risks of thrombolysis by clinicians to patients/relatives. Using mixed methods we developed a COMPuterised decision Aid for Stroke thrombolysiS (COMPASS) in an iterative staged process (review of available tools; a decision analytic model; interactive group workshops with clinicians and patients/relatives; and prototype usability testing). We then tested the tool in simulated situations with final testing in real life stroke thrombolysis decisions in hospitals. Clinicians used COMPASS pragmatically in managing acute stroke patients potentially eligible for thrombolysis; their experience was assessed using self-completion forms and interviews. Computer logged data assessed time in use, and utilisation of graphical risk presentations and additional features. Patients'/relatives' experiences of discussions supported by COMPASS were explored using interviews. COMPASS expresses predicted outcomes (bleeding complications, death, and extent of disability) with and without thrombolysis, presented numerically (percentages and natural frequencies) and graphically (pictographs, bar graphs and flowcharts). COMPASS was used for 25 patients and no adverse effects of use were reported. Median time in use was 2.8 minutes. Graphical risk presentations were shared with 14 patients/relatives. Clinicians (n = 10) valued the patient-specific predictions of benefit from thrombolysis, and the support of better risk communication with patients/relatives. Patients (n = 2) and relatives (n = 6) reported that graphical risk presentations facilitated understanding of benefits/risks of thrombolysis. Additional features (e.g. dosage calculator) were suggested and subsequently embedded within COMPASS to enhance usability. Our structured development process led to the development of a gamma prototype computerised decision aid. Initial evaluation has demonstrated reasonable acceptability of COMPASS amongst patients, relatives and clinicians. The impact of COMPASS on clinical outcomes requires wider prospective evaluation in clinical settings.


Engineering acetyl-CoA metabolic shortcut for eco-friendly production of polyketides triacetic acid lactone in Yarrowia lipolytica.
Acetyl-CoA is the central metabolic node connecting glycolysis, Krebs cycle and fatty acids synthase. Plant-derived polyketides, are assembled from acetyl-CoA and malonyl-CoA, represent a large family of biological compounds with diversified bioactivity. Harnessing microbial bioconversion is considered as a feasible approach to large-scale production of polyketides from renewable feedstocks. Most of the current polyketide production platform relied on the lengthy glycolytic steps to provide acetyl-CoA, which inherently suffers from complex regulation with metabolically-costly cofactor/ATP requirements. Using the simplest polyketide triacetic acid lactone (TAL) as a testbed molecule, we demonstrate that acetate uptake pathway in oleaginous yeast (Yarrowia lipolytica) could function as an acetyl-CoA shortcut to achieve metabolic optimality in producing polyketides. We identified the metabolic bottlenecks to rewire acetate utilization for efficient TAL production in Y. lipolytica, including generation of the driving force for acetyl-CoA, malonyl-CoA and NADPH. The engineered strain, with the overexpression of endogenous acetyl-CoA carboxylase (ACC1), malic enzyme (MAE1) and a bacteria-derived cytosolic pyruvate dehydrogenase (PDH), affords robust TAL production with titer up to 4.76 g/L from industrial glacier acetic acid in shake flasks, representing 8.5-times improvement over the parental strain. The acetate-to-TAL conversion ratio (0.149 g/g) reaches 31.9% of the theoretical maximum yield. The carbon flux through this acetyl-CoA metabolic shortcut exceeds the carbon flux afforded by the native glycolytic pathways. Potentially, acetic acid could be manufactured in large-quantity at low-cost from Syngas fermentation or heterogenous catalysis (methanol carbonylation). This alternative carbon sources present a metabolic advantage over glucose to unleash intrinsic pathway limitations and achieve high carbon conversion efficiency and cost-efficiency. This work also highlights that low-cost acetic acid could be sustainably upgraded to high-value polyketides by oleaginous yeast species in an eco-friendly and cost-efficient manner.


Natural infection of humans, animals, and phlebotomine sand flies with the Alagoas serotype of vesicular stomatitis virus in Colombia.
Five isolations of the Alagoas serotype of vesicular stomatitis virus (Rhabdoviridae: Vesiculovirus) were made from naturally infected phlebotomine sand flies (Lutzomyia spp.) collected in Colombia. These are the first isolations of Alagoas virus from an arthropod. Replication of the virus occurred in laboratory-reared sand flies (Lutzomyia longipalpis) after inoculation. Bite and transovarial transmission of the virus was also demonstrated in experimentally infected sand flies. Alagoas virus neutralizing antibodies were found in sera of humans and animals living near the insect collection site; antibody rates among human residents of two nearby towns were 63% and 83%, respectively. Results of comparative serologic studies demonstrated that Alagoas virus is closely related antigenically to Indiana, Cocal, and Maraba viruses and that these four agents form a complex within the vesicular stomatitis virus serogroup. The antigenic similarity among these four viruses makes their differentiation difficult; it also raises doubts about the accuracy of current laboratory methods used for identifying isolates in this serogroup. A discussion follows on the significance of human antibodies to these agents and on the role of sand flies in their ecology.


The role of calcium and non calcium-based phosphate binders in chronic kidney disease.
Rising levels of serum phosphate occur late in the course of chronic kidney disease (CKD) and have been easy targets for nephrologists to treat using phosphate binding drugs, as well as fertile ground for the pharmaceutical industry, for meta-analysis and for the earnest pontifications of guideline writers. Unfortunately, the evidence does little to support this focus, which might be better applied to earlier, adaptive hormonal changes, and to phosphate balance rather than serum phosphate levels. Nevertheless, phosphate binders are ubiquitously prescribed to patients on dialysis, and often prescribed to patients with earlier stages of CKD; for which there is no evidence of benefit and some evidence that calcium-based binders (CBBs) and possibly non-CBBs may cause more harm than placebo. For patients on dialysis, observational studies suggest that phosphate binder use may reduce mortality. Meta-analyses report reduced hypercalcaemia for non-CBBs versus CBBs, and compared with CBBs sevelamer has been reported to significantly reduce vascular calcification progression and, in some but not all studies, overall mortality. However, limitations include study heterogeneity. For patients on dialysis, CBBs appear to have no advantages over the non-CBBs, apart from lower cost. In general, it seems prudent to avoid prescribing calcium-based drugs to patients who are pre-dialysis, and to use non-CBBs preferentially in CKD stage 5D. Current draft Kidney Disease Improving Global Outcomes guidelines suggest that CBB dosage be minimized in CKD stages 3 to 5D.


Feasibility of SHARK: a physical activity skill-development program for overweight and obese children.
This study aimed to assess the feasibility of a community-based physical activity motor development program, called SHARK, among overweight and obese children. Thirteen overweight or obese children aged 8-12 years were recruited to the 10-week program from the Illawarra region of NSW, Australia. Measures at baseline, post-treatment and 9-month follow-up included BMI, motor development, perceived competence, objectively measured physical activity and performance of an activity of daily living. At post-treatment and follow-up motor development, perceived athletic competence and perceived global self-worth had significantly increased. Although recorded minutes in moderate-to-vigorous activity significantly declined at post-treatment and follow-up, activity daily counts did not. The results indicate that community-based activity programs designed to improve the motor development and perceived competence of overweight and obese children are feasible and may have important and sustainable benefits. However, a larger sample size and a comparison group are required to determine whether this approach can impact on activity levels and weight status.


Interacting noise sources shape patterns of arm movement variability in three-dimensional space.
Reaching movements are subject to noise in both the planning and execution phases of movement production. The interaction of these noise sources during natural movements is not well understood, despite its importance for understanding movement variability in neurologically intact and impaired individuals. Here we examined the interaction of planning and execution related noise during the production of unconstrained reaching movements. Subjects performed sequences of two movements to targets arranged in three vertical planes separated in depth. The starting position for each sequence was also varied in depth with the target plane; thus required movement sequences were largely contained within the vertical plane of the targets. Each final target in a sequence was approached from two different directions, and these movements were made with or without visual feedback of the moving hand. These combined aspects of the design allowed us to probe the interaction of execution and planning related noise with respect to reach endpoint variability. In agreement with previous studies, we found that reach endpoint distributions were highly anisotropic. The principal axes of movement variability were largely aligned with the depth axis, i.e., the axis along which visual planning related noise would be expected to dominate, and were not generally well aligned with the direction of the movement vector. Our results suggest that visual planning-related noise plays a dominant role in determining anisotropic patterns of endpoint variability in three-dimensional space, with execution noise adding to this variability in a movement direction-dependent manner.


Limitations of Photosynthesis in Pinus taeda L. (Loblolly Pine) at Low Soil Temperatures.
The relative importance of stomatal and nonstomatal limitations to net photosynthesis (A) and possible signals responsible for stomatal limitations were investigated in unhardened Pinus taeda seedlings at low soil temperatures. After 2 days at soil temperatures between 13 and 7 degrees C, A was reduced by 20 to 50%, respectively. The reduction in A at these moderate root-chilling conditions appeared to be the result of stomatal limitations, based on the decrease in intercellular CO(2) concentrations (c(i)). This conclusion was supported by A versus c(i) analysis and measurements of O(2) evolution at saturating CO(2), which suggested increases in stomatal but not biochemical limitations at these soil temperatures. Nonuniform stomatal apertures, which were demonstrated with abscisic acid, were not apparent 2 days after root chilling, and results of our A versus c(i) analysis appear valid. Bulk shoot water potential (psi) declined as soil temperature dropped below 16 degrees C. When half the root system of seedlings was chilled, shoot psi and gas-exchange rates did not decline. Thus, nonhydraulic root-shoot signals were not implicated in stomatal limitations. The initial decrease in leaf conductance to water vapor after root chilling appeared to precede any detectable decrease in bulk fascicle psi, but may be in response to a decrease in turgor of epidermal cells. These reductions in leaf conductance to water vapor, which occurred within 30 minutes of root chilling, could be delayed and temporarily reversed by reducing the leaf-to-air vapor-pressure deficit, suggesting that hydraulic signals may be involved in initiating stomatal closure. By independently manipulating the leaf-to-air vapor-pressure deficit of individual fascicles, we could induce uptake of water vapor through stomata, suggesting that nonsaturated conditions occur in the intercellular airspaces. There was an anomaly in our results on seedlings maintained for 2 days at soil temperatures below 7 degrees C. Lower A appeared primarily the result of nonstomatal limitations, based on large increases in calculated c(i) and A versus c(i) analysis. In contrast, measurements of O(2) evolution at saturating CO(2) concentrations implied nonstomatal limitations per se did not increase at these temperatures. One explanation for this paradox is that calculations of c(i) are unreliable at very low gas-exchange rates because of inadequate measurement resolution, and limitations of A are predominantly stomatal. An alternative interpretation is that increases in c(i) are real and the results from O(2)-evolution measurements are in error. The high CO(2) concentration used in O(2)-evolution measurements (15%) may have overcome nonstomatal limitations by enzymes that were down-regulated by a feedback mechanism. In this scenario, carbohydrate feedback limitations may be responsible for nonstomatal reductions in A after 2 days at soil temperatures below 7 degrees C.


Syntheses of C17-C27 fragments of 20-deoxybryostatins for assembly using Julia and metathesis reactions.
Two approaches to the synthesis of compounds corresponding to the C17-C27 fragment of the 20-deoxybryostatins are described. The first approach is based on the palladium(0) catalysed coupling of tin enolates, generated in situ from enol acetates using tributyltin methoxide, with vinylic bromides. The vinylic bromides were prepared using the Sharpless asymmetric dihydroxylation to introduce the hydroxyl groups corresponding to those at C25 and C26 in the bryostatins. Following several steps to introduce alkynyl ester functionality, the stereoselective addition of a tributyltin cuprate followed by tributyltin-bromine exchange gave the required vinylic bromides. The palladium(0) catalysed couplings worked very well for enol esters containing thioether substituents and gave products with retention of the position and geometry of the trisubstituted double bond derived from the vinylic bromide. These were taken through to compounds corresponding to fully developed C17-C27 fragments ready for assembly of the 16,17-double-bond of bryostatins by Julia reactions. This chemistry was also applied to prepare intermediates suitable for incorporation into bryostatins by ring-closing metathesis but, in this case, the coupling reaction gave mixtures of products including both the required βγ-unsaturated ketone and a conjugated diene formed by a competing Heck reaction. To avoid this problem, a second approach to compounds suitable for incorporation into a metathesis-based assembly of 20-deoxybryostatins was developed. In this organotin-free synthesis, the key step was the conjugate addition of an organic cuprate generated from allylmagnesium bromide to an alkynoate that gave the required (Z)-trisubstituted alkene with excellent stereoselectivity. This was converted into metathesis precursors in a few steps.


Phase I trial of intraperitoneal iododeoxyuridine with and without intravenous high-dose folinic acid in the treatment of advanced malignancies primarily confined to the peritoneal cavity: flow cytometric and pharmacokinetic analysis.
In this Phase I study, the maximally tolerated doses (MTDs) of i.p. iododeoxyuridine (IdUrd) alone and in combination with i.v. calcium leucovorin (LV) were determined. The pharmacokinetics and pharmacological advantage of IdUrd were evaluated, and flow cytometric analysis allowed examination of the extent of incorporation of IdUrd into tumor cells with and without the addition of i.v. LV. Thirty-nine patients with advanced neoplasms primarily confined to the peritoneal space were enrolled in a dose-escalation trial using 4-h dwells of IdUrd administered i.p. daily for 4 days with and without an i.v. infusion of LV 500 mg/m2/day for 4.5 days. Twenty-three patients received single-agent therapy, and 13 patients received i.p. IdUrd in combination with i.v. LV. The MTD of single-agent IdUrd administered on this schedule was 4125 mg/m2/day for 4 days; and that of the IdUrd in combination was 3438 mg/m2/day. Dose-limiting toxicities were myelosuppression and stomatitis. During the period of the dwell, the peritoneal AUC (area under the curve) of IdUrd exceeded the plasma AUC of IdUrd by one or two orders of magnitude in all patients at all doses tested; there was a possible effect of LV on peritoneal AUC. The geometric mean pharmacological advantage (AUCperitoneal/ AUCplasma) was 181 at 625 mg/m2/day and 90 at 4538 mg/m2/day. Flow cytometric analysis suggests saturation of IdUrd measured in DNA at the 2500-3125 mg/m2 dose level, without an increase after the addition of LV. Twelve patients received 4-12 courses of therapy. One patient with recurrent ovarian cancer who received 16 courses of therapy experienced complete resolution of her ascites, near normalization of CA-125 levels, and improved quality of life; two patients with high-risk tumors receiving "adjuvant" therapy are disease-free at 3 and 6 years after treatment; other patients experienced transient clearing of ascites. The recommended Phase II dose of i.p. IdUrd using a 4-h dwell daily for 4 days is 3750 mg/m2/day alone or 3125 mg/m2/day in combination with continuous i.v. LV at 500 mg/m2/day for 4.5 days. Although flow cytometric data suggest that DNA incorporation of IdUrd is not affected by the addition of LV, the cytotoxicity of the combination regimen may be increased due to LV-enhanced, IdUrd-related inhibition of thymidylate synthase. For this reason, we recommend that efficacy studies of the combination continue in parallel with studies of IdUrd alone.


Dose escalation study of high-dose carboplatin and etoposide with autologous bone marrow support in patients with recurrent and refractory germ cell tumors.
Thirty-three patients with germ cell cancer (GCT) recurrent after two cisplatin-based regimens or cisplatin refractory (progression within 4 weeks of the last dose of cisplatin) were enrolled in a trial to establish the maximum tolerated doses (MTD) of carboplatin and etoposide given in combination with ABMT for two cycles. BM harvest of > or = 2 x 10(8) nucleated cells/kg preceded two cycles of therapy. Each agent was dose escalated, carboplatin from 1650 mg/m2 to 2100 mg/m2 and etoposide from 1200 mg/m2 to 2250 mg/m2 per cycle in successive cohorts. Twenty patients completed two cycles, 13 underwent only one due to: early death (4), toxicity (2), and progressive disease (6). There were four CR, three of whom achieved NED status with surgery, 14 PR, of whom eight have progressed. Four patients with stable disease and seven PD have died with a median survival of 6 months. There were six treatment-related deaths, four on course 1 and two on course 2. Causes of death on course 1 were: CNS hemorrhage (1), multiorgan failure (3); and on course 2: sepsis (1) and sudden death (1). Severe but reversible mucositis, transaminase and creatinine elevations were observed at the highest dose level. Three of five patients treated at this dose level had severe neurologic toxicity, manifested by both peripheral neuropathy and ototoxicity. The MTD in this patient population was carboplatin 2100 mg/m2 and etoposide 2250 mg/m2 on each of two cycles of therapy. Neurologic and mucosal toxicity were dose limiting.


Prognostic Value of MTV, SUVmax and the T/N Ratio of PET/CT in Patients with Glioma: A Systematic Review and Meta-Analysis.
Background: In the past decade, positron emission tomography/computed tomography (PET/CT) has become an important imaging tool for clinical assessment of tumor patients. Our meta-analysis aimed to compare the predictive value of PET/CT parameters regard to overall survival (OS) and progression-free survival (PFS) outcomes in glioma. Methods: Relevant articles were systematically searched in PMC, PubMed, EMBASE and WEB of science. Studies involving the prognostic roles of PET/CT parameters with OS and PFS in glioma patients were evaluated. The impact of metabolic tumor volume (MTV), maximal standard uptake value (SUVmax), and the ratio of uptake in tumor to normal (T/N ratio) on survival was measured by calculating combined hazard ratios (HRs) and 95% confidence intervals (CIs). Results: A total of 32 articles with 1715 patients were included. The combined HRs of higher MTV, higher SUVmax and higher T/N ratio for OS were 1.14 (95% CI: 0.98-1.32, P heterogeneity<0.001), 1.69 (95% CI: 1.18-2.41, P heterogeneity<0.001) and 1.68 (95% CI: 1.40-2.01, P heterogeneity< 0.001), respectively. Regarding PFS, the combined HRs were 1.04 (95% CI: 0.97-1.11, P heterogeneity=0.002) with higher MTV, 1.45 (95% CI: 1.11-1.90, P heterogeneity<0.001) with higher SUVmax and 2.07 (95% CI: 1.45-2.95, P heterogeneity<0.001) with higher T/N ratio. Results remained similar in the sub-group analyses. Conclusion: PET/CT parameters T/N ratio may be a significant prognostic factor in patients with glioma. Evidence of SUVmax and MTV needed more large-scale studies performed to validate. PET/CT scan could be a promising technique to provide prognostic information for these patients.


Identification of pain indicators for infants at risk for neurological impairment: a Delphi consensus study.
A number of infant pain measures have been developed over the past 15 years incorporating behavioural and physiologic indicators; however, no reliable or valid measure exists for infants who are at risk for neurological impairments (NI). The objective of this study was to establish consensus about which behavioural, physiologic and contextual indicators best characterize pain in infants at high, moderate and low levels of risk for NI. A 39- item, self-administered electronic survey that included infant physiologic, behavioral and contextual pain indicators was used in a two round Delphi consensus exercise. Fourteen pediatric pain experts were polled individually and anonymously on the importance and usefulness of the pain indicators for the 3 differing levels of risk for NI. The strength of agreement between expert raters was moderate in Round 1 and fair in Round 2. In general, pain indicators with the highest concordance for all three groups were brow bulge, facial grimace, eye squeeze, and inconsolability. Increased heart rate from baseline in the moderate and severe groups demonstrated high concordance. In the severe risk group, fluctuations in heart rate and reduced oxygen saturation were also highly rated. These data constitute the first step in contributing to the development and validation of a pain measure for infants at risk for NI. In future research, we will integrate these findings with the opinions of (a) health care providers about the importance and usefulness of infant pain indicators and (b) the pain responses of infants at mild, moderate and high risk for NI.


Implication of protein kinase C in IL-2-mediated proliferation and apoptosis in a murine T cell clone.
Several reports have suggested that protein kinase C plays an essential role in T cell activation and apoptosis. The recent synthesis of the selective protein kinase C inhibitor, GF109203X, has made it possible to test the relevance of protein kinase C in T cell proliferation and apoptosis. We report that the use of GF109203X, in concentrations that are below its toxicity limits, inhibits IL-2-dependent proliferation in murine T cells expressing intermediate or high-affinity IL-2R (TS1 beta and TS1 alpha beta). In addition, GF109203X reverts the suppression of apoptosis mediated by IL-2 or IL-2+ dexamethasone. The use of the phorbol ester PMA, a protein kinase C activator, allows a bypass of the IL-2/IL-2R interaction in the suppression of apoptosis mediated by dexamethasone or IL-2 withdrawal in TS1 beta cells but not in TS1 alpha beta cells. Taken together, our data indicate that activation of protein kinase C is an important step in IL-2-mediated proliferation and in suppression of apoptosis.


Comparison of lecture and team-based learning in medical ethics education.
Medical education literature suggests that ethics education should be learner-centered and problem-based rather than theory-based. Team-based learning is an appropriate method for this suggestion. However, its effectiveness was not investigated enough in medical ethics education. Is team-based learning effective in medical ethics education in terms of knowledge retention, in-class learner engagement, and learner reactions? This was a prospective controlled follow-up study. We changed lecture with team-based learning method to teach four topics in a 2-week medical ethics clerkship, while the remaining topics were taught by lectures. For comparison, we formed team-based learning and lecture groups, in which the students and instructor are the same, but the topics and teaching methodologies are different. We determined in-class learner engagement by direct observation and student satisfaction by feedback forms. Student success for team-based learning and lecture topics in the end-of-clerkship exam and two retention tests performed 1 year and 2 years later were compared. Ethical approval for the study was granted by Akdeniz University Board of Ethics on Noninvasive Clinical Human Studies Ethics committee. Short-term knowledge retention did not differ; however, team-based learning was found superior to lecture at long-term retention tests. Student satisfaction was high with team-based learning and in-class engagement was better in team-based learning sessions. Our results on learner engagement and satisfaction with team-based learning were similar to those of previous reports. However, knowledge retention results in our study were contrary to literature. The reason might be the fact that students prepared for the end-of-clerkship pass/fail exam (short term) regardless of the teaching method. But, at long-term retention tests, they did not prepare for the exam and answered the questions just using the knowledge retained in their memories. Our findings suggest that team-based learning is a better alternative to lecture to teach ethics in medical education.


The effects of menstrual cycle on the knee joint position sense: preliminary study.
The purpose of this study was to determine the effects of menstrual cycle on proprioception by using the active knee joint position sense test (JPST). The 19 healthy women (ages between 20 years and 27 years) who have normal regular menstrual cycle were included in the study. We applied JPSTs at two different directions throughout the three different phases of the menstrual cycle, i.e. menstrual, follicular, and early luteal in dominant knees. When we started from flexion (90 degrees ), target angles were 70 degrees , 50 degrees , and 30 degrees and we started from extension (0 degrees ), target angles were 20 degrees , 40 degrees and 60 degrees . The absolute reposition errors from the target angles have been evaluated. Results have shown that reposition errors from the target angle at 40 degrees , 50 degrees and 70 degrees of knee angles were higher in the menstrual phase than that of the follicular phase (P<0.05). In addition, higher value of reposition error from the target angle at 40 degrees was found in the menstrual phase compared to luteal phase (P<0.05). In conclusion, we have demonstrated that active JPST was significantly reduced in the menstruation period.


A phase 2 study of vorinostat in acute myeloid leukemia.
This two-stage, multi-institutional, randomized phase 2 trial assessed the toxicity and response rate associated with two treatment schedules of the histone deacetylase inhibitor, vorinostat (suberoylanilide hydroxamic acid; SAHA) in patients with relapsed acute myeloid leukemia and in selected untreated patients with high-risk acute myeloid leukemia. Patients with relapsed or untreated acute myeloid leukemia who were not candidates for chemotherapy entered one of the two treatment arms. In both arms a total dose of 8400 mg of vorinostat was delivered in each 21-day cycle of treatment: in arm A the dose regimen was 400 mg daily whereas in arm B the dose regimen was 200 mg three times daily for 14 days followed by 1 week rest. Data from all 37 patients were used for the analyses. In arm A (n=15), the confirmed complete remission rate was 0% (95% CI, 0% to 23%); this arm was closed at the planned interim analysis. In arm B (n=22), the confirmed complete remission rate was 4.5% (1 response; 95% CI, 0.4% to 24%), with a duration of response exceeding 398 days. The median time to treatment failure in arm A was 42 days (95% CI, 26 to 57); although a minimum of four cycles of treatment were planned, 11 patients (79%) received no more than two cycles. The median time to treatment failure in arm B was 46 days (95% CI, 20 to 71); 13 patients (59%) received no more than two cycles of treatment. Vorinostat monotherapy demonstrated minimal activity in this group of patients with acute myeloid leukemia. Therapy was discontinued in many patients before the planned four cycles had been administered, either because of failure of vorinostat to control the leukocyte count or patients' and physicians' preference. Future studies of vorinostat in acute myeloid leukemia should focus on combinations with other drugs with which it might interact pharmacodynamically. ClinicalTrials.gov Identifier: NCT00305773.


Incidence, etiology and predictors of adverse outcomes in 43,315 patients presenting to the Emergency Department with syncope: an international meta-analysis.
Syncope remains challenging for Emergency Department (ED) physicians due to difficulties in assessing the risk of future adverse outcomes. The aim of this meta-analysis is to establish the incidence and etiology of adverse outcomes as well as the predictors, in patients presenting with syncope to the ED. A systematic electronic literature review was performed looking for eligible studies published between 1990 and 2010. Studies reporting multivariate predictors of adverse outcomes in patients presenting with syncope to the ED were included and pooled, when appropriate, using a random-effect method. Adverse events were defined as 'incidence of death, or of hospitalization and interventional procedures because of arrhythmias, ischemic heart disease or valvular heart disease'. 11 studies were included. Pooled analysis showed 42% (CI 95%; 32-52) of patients were admitted to hospital. Risk of death was 4.4% (CI 95%; 3.1-5.1) and 1.1% (CI 95%; 0.7-1.5) had a cardiovascular etiology. One third of patients were discharged without a diagnosis, while the most frequent diagnosis was 'situational, orthostatic or vasavagal syncope' in 29% (CI 95%; 12-47). 10.4% (CI 95%; 7.8-16) was diagnosed with heart disease, the most frequent type being bradyarrhythmia, 4.8% (CI 95%; 2.2-6.4) and tachyarrhythmia 2.6% (CI 95%; 1.1-3.1). Palpitations preceding syncope, exertional syncope, a history consistent of heart failure or ischemic heart disease, and evidence of bleeding were the most powerful predictors of an adverse outcome. Syncope carries a high risk of death, mainly related to cardiovascular disease. This large study which has established the most powerful predictors of adverse outcomes, may enable care and resources to be better focused at high risk patients.


Quantitation of spinal cord demyelination, remyelination, atrophy, and axonal loss in a model of progressive neurologic injury.
Spinal cord pathology, such as demyelination and axonal loss, is a common feature in multiple models of central nervous system (CNS) injury and disease. Development of methods to quantify spinal cord pathology objectively would aid studies designed to establish mechanisms of damage, correlate pathology with neurologic function, and assess therapeutic interventions. In this study, we describe sensitive methods to objectively quantify spinal cord demyelination, remyelination, atrophy, and axonal loss following the initiation of a progressive inflammatory demyelinating disease with Theiler's murine encephalomyelitis virus (TMEV). Spinal cord demyelination, remyelination, and atrophy were quantified from representative 1-microm-thick cross sections embedded in Araldite plastic using interactive image analysis. In addition, this study demonstrates novel, automated methodology to quantify axonal loss from areas of normal-appearing white matter, as a measure of secondary axonal injury following demyelination. These morphologic methods, which are applicable to various models of CNS injury, provide an innovative way to assess the benefits of therapeutic agents, to determine mechanisms of spinal cord damage, or to establish a correlation with sensitive measures of neurologic function. J. Neurosci Res 58:492-504.


The grana of starch-free chloroplasts of Nicotiana rustica.
The grana of chloroplasts of starch-free leaves of Nicotiana rustica are described in detail. Leaf sections were fixed in 2.5 per cent KMnO(4) and embedded in mixtures of butyl and ethyl methacrylate. Chain length of the polymer was modified by use of a transfer agent. The grana are composed of compartments consisting of electron-scattering partitions and electron-transparent loculi. Compartments are not open to the stroma so that the grana are distinct subplastid organelles. Adjacent grana are connected by an anastomosing fretwork system composed of flexuous channels bordered by electron-scattering membranes. Ten different kinds of granum margins are described. These marginal variations depend upon grana-fretwork connections and internal marginal connections between adjacent loculi. A study of serial sections suggests, at least in some plastids, the occurrence of a possible orderly spiral arrangement of compartment-fretwork connections. Adjacent grana may have common compartments. Grana may branch. Variations in shape may depend upon the angle the section bears to the axis of the cylinder. This should also influence the relative thickness and sharpness of the partitions. Since all shapes and variations in partition thickness and sharpness cannot be accounted for on the basis of the orientation of the grana, such variations probably occur naturally. Grana vary in size, ranging from those few which have a single partition to those having 50 or more compartments which extend completely through the width of a plastid. Relationships between grana of different sizes and between compartments and frets indicate the possibility of growth of grana from union or extension of compartments and formation of compartments from the union of frets.


[Consequences of cocaine use among prostitutes].
To describe the circumstances and consequences of cocaine use in the setting of prostitution. We performed a qualitative study with fieldwork in 6 Spanish cities. Semi-structured interviews were carried out to 60 persons aged between 18 and 50 years old involved in prostitution in different settings: street, flat and brothel. The main substances used in prostitution were alcohol and cocaine. Consumption was occasional and instrumental and helped to reduce psychological barriers or inhibition and increase tolerance to long hours of sex work with different clients, thus increasing earnings. Cocaine use was also associated with providing clients with company without having to perform sexual services. Among the consequences of drug use in prostitutes were unprotected sex and a higher risk of violence by clients. Prostitutes developed harm reduction strategies for drug use: dose reduction, simulated drug use, and selection of clients who were users. This study shows that there is connection between drug use and the provision of sexual services. Drug use can pose a health risk as consumption is associated with unprotected sex.


The impact of the COVID-19 pandemic on marginalized populations in the United States: A research agenda.
International and national crises often highlight inequalities in the labor market that disproportionately affect individuals from marginalized backgrounds. The COVID-19 pandemic, and the resulting changes in society due to social distancing measures, has showcased inequities in access to decent work and experiences of discrimination resulting in many of the vulnerable populations in the United States experiencing a much harsher impact on economic and work-related factors. The purpose of this essay is to describe how the COVID-19 pandemic may differentially affect workers of color, individuals from low-income backgrounds, and women in complex ways. First, this essay will discuss disproportionate representation of workers from low-income and racial/ethnic minority backgrounds in sectors most affected by COVID-19. Second, it will discuss the lack of decent work for low-income workers who perform "essential" tasks. Third, this essay will highlight economic and work-related implications of increased discrimination Asian Americans are experiencing in society. Finally, role conflict and stress for women who are managing additional unpaid work, including caretaking responsibilities, while needing to continue to engage in paid work will be examined. A research agenda will be set forth throughout the essay, calling for vocational psychologists to engage in research that fully examines how the COVID-19 pandemic is affecting vulnerable communities.


Psychopathology and coping in recently diagnosed HIV/AIDS patients--the role of gender.
Although there is growing literature on the psychological responses to and the psychopathology associated with HIV/AIDS, few investigations have focused on the role of gender. This study compared psychiatric morbidity, coping responses, and disability in male and female outpatients recently diagnosed with HIV/AIDS. One hundred and forty-nine patients (44 male, 105 female) with HIV/AIDS (mean +/- standard deviation (SD) months since diagnosis 5.8 +/- 4.1) attending an infectious diseases clinic at Tygerberg Hospital, Cape Town, were evaluated. Subjects were assessed using the MINI International Neuropsychiatric Interview (MINI), the Carver Brief COPE, and the Sheehan Disability Scale. In addition, negative life events and risk behaviours were evaluated. Fifty-six per cent of patients were diagnosed with a psychiatric disorder, most commonly major depression (34.9%), dysthymic disorder (21.5%), post-traumatic stress disorder (14.8%), and alcohol dependence (10.1%). There were no significant gender differences in the prevalence of mood disorders in the sample. Men, however, were more likely than women to meet diagnostic criteria for alcohol abuse or dependence, and to engage in certain risky sexual behaviours. Women were more likely to suffer from post-traumatic stress disorder, and to use coping strategies of planning and religion to deal with the illness. There were no significant gender differences in disability. Psychiatric disorders are common in recently diagnosed HIV/AIDS patients in South Africa. Clinicians should be aware of the high prevalence of mood disorders in both men and women, and of gender-different responses such as increased alcohol and substance use and more risky sexual behaviour in men.


Interferon-gamma inhibits healing post scald burn injury.
Impaired healing after severe burns remains a reason for prolonged hospitalization, opportunistic infections, and debilitating scarring. Interferon-gamma (IFN-γ) is an important immune regulator that has been shown to inhibit collagen synthesis by fibroblasts, resulting in delayed healing in incision wounds. To determine whether IFN-γ plays similar roles in the healing process after severe burn, we induced scald injury in mice deficient or sufficient in IFN-γ and examined local responses. In the absence of IFN-γ, scalded areas healed faster. This was associated with attenuated local inflammatory responses, enhanced reepithelialization, increased proliferation of keratinocytes in reepithelialized leading edges, and up-regulation of growth factors in burned skin areas. Furthermore, angiogenesis and myofibroblast formation commenced and terminated earlier in IFN-γ(-/-) mice compared with wild type (WT) controls. Our observations demonstrate that inhibition of IFN-γ results in accelerated healing after burn injury by dampening excessive inflammation and facilitating reepithelialization, collagen deposition, and wound contraction.


Effect of Moringa oleifera on hematological parameters of calves reared in industrial fluorotic area.
The present study was undertaken to evaluate the ameliorative potential of dried Moringa oleifera fruit powder in fluorosis affected calves reared around the vicinity of aluminium smelter plant. Total 107 calves were screened on the basis of clinical signs and higher plasma fluoride (more than 0.2 ppm) level for evidence of fluorosis. Out of that, 90 samples found positive and from them 18 calves of 6-12 months age group were selected and divided equally into three groups named as Group II, III, and IV. Group II remained as disease control group whereas Group III calves were supplemented with dried M. oleifera fruit powder of 25 g/calve for 60 days. Group IV calves were supplemented with calcium carbonate at 100 mg/kg body weight and boric acid at 10 mg/kg for the same experimental period. Group I consisted of six numbers of healthy calves taken from the non-fluorotic zone, i.e. Bhubaneswar. Plasma fluoride level, hemoglobin (Hb), packed cell volume (PCV), total leukocyte count (TLC), differential count (DC), total erythrocyte count, mean corpuscular volume (MCV), mean corpuscular Hb (MCH), and MCH concentration (MCHC) were estimated on day 0, 30, and 60 of the experiment. Supplementation of dried M. oleifera fruit powder to fluorosis affected calves resulted in significant reduction in plasma fluoride level and increase in Hb%, PCV, TLC and altered DC. Similar results were also recorded in calcium+boron group, except PCV and Hb. No significant changes were observed in MCV, MCH, and MCHC values. The present study concluded that supplementation of dried M. oleifera fruit powder daily for 60 days has shown protection against chronic fluoride toxicity in calves.


Sensory overresponsivity and anxiety in children with ADHD.
Approximately 25% of children with attention deficit hyperactivity disorder (ADHD) have a comorbid anxiety disorder. The purpose of this study was to determine whether sensory overresponsivity (SOR) is related to elevated levels of anxiety in children with ADHD. Twenty-four children ages 6 to 10 with ADHD and 24 children without ADHD participated in this study. All parentscompleted a Revised Children's Manifest Anxiety Scale (RCMAS) with their child. Children in the ADHD group were subdivided into ADHD + SOR and ADHD-only groups using the Sensory Over-Responsivity Inventory. Children in the ADHD + SOR group were significantly more anxious than both the ADHD-only and non-ADHD (control) groups. Children with ADHD + SOR were also more likely to have clinically significant anxiety (determined by total scores on the RCMAS). Occupational therapists treating children with ADHD and SOR should be aware that these children may also have anxiety and discuss options with families for prevention or treatment.


Evaluating Clinical Isolates for Their Phenotypic and Genotypic Resistance Against Anti-HIV Drugs.
The high replication rate of HIV, together with the low fidelity of its reverse transcriptase, provides the virus with an unprecedented genomic flexibility. This allows a fast adaptation to selective pressure, including antiviral drugs, resulting in the development of drug-resistant strains. The present improvements in the treatment of AIDS patients are at least partly owing to antiviral therapy. To assess the implications of HIV drug resistance on patient management, drug resistance assays for clinical HIV isolates are widely being used. Ideally, monitoring drug resistance should help clinicians in their treatment decisions. If patients would really benefit clinically from this strategy, then the gain from clinical improvement and from omitting drugs to which the virus is already resistant would outweigh the cost of drug resistance testing. In the next few years, researchers should consolidate the clinical benefit of antiviral drug resistance testing, and for this they need fast, reliable and cheap assays. All present assays are in vitro assays, which can only partly mimic the in vivo situation with confounding factors, such as cellular resistance (1). Efforts are presently made to establish in vivo assays (2; see also Chapter 10 ).


In vitro studies on colonization resistance of the human gut microbiota to Candida albicans and the effects of tetracycline and Lactobacillus plantarum LPK.
An anaerobic three-vessel continuous-flow culture system, which models the three major anatomical regions of the human colon, was used to study the persistence of Candida albicans in the presence of a faecal microbiota. During steady state conditions, overgrowth of C. albicans was prevented by commensal bacteria indigenous to the system. However antibiotics, such as tetracycline have the ability to disrupt the bacterial populations within the gut. Thus, colonization resistance can be compromised and overgrowth of undesirable microorganisms like C. albicans can then occur. In this study, growth of C. albicans was not observed in the presence of an established faecal microbiota. However, following the addition of tetracycline to the growth medium, significant growth of C. albicans occurred. A probiotic Lactobacillus plantarum LPK culture was added to the system to investigate whether this organism had any effects upon the Candida populations. Although C. albicans was not completely eradicated in the presence of this bacterium, cell counts were markedly reduced, indicating a compromised physiological function. This study shows that the normal gut flora can exert 'natural' resistance to C. albicans, however this may be diminished during antibiotic intake. The use of probiotics can help fortify natural resistance.


Association of Overlapping Surgery With Increased Risk for Complications Following Hip Surgery: A Population-Based, Matched Cohort Study.
Overlapping surgery, also known as double-booking, refers to a controversial practice in which a single attending surgeon supervises 2 or more operations, in different operating rooms, at the same time. To determine if overlapping surgery is associated with greater risk for complications following surgical treatment for hip fracture and arthritis. This was a retrospective population-based cohort study in Ontario, Canada (population, 13.6 million), for the years 2009 to 2014. There was 1 year of follow-up. This study encompassed 2 large cohorts. The "hip fracture" cohort captured all persons older than 60 years who underwent surgery for a hip fracture during the study period. The "total hip arthroplasty" (THA) cohort captured all primary elective THA recipients for arthritis during the study period. We matched overlapping and nonoverlapping hip fractures by patient age, patient sex, surgical procedure (for the hip fracture cohort), primary surgeon, and hospital. Procedures were identified as overlapping if they overlapped with another surgical procedure performed by the same primary attending surgeon by more than 30 minutes. Complication (infection, revision, dislocation) within 1 year. There were 38 008 hip fractures, and of those, 960 (2.5%) were overlapping (mean age of patients, 66 years [interquartile range, 57-74 years]; 503 [52.4%] were female). There were 52 869 THAs and of those, 1560 (3.0%) overlapping (mean age, 84 years [interquartile range, 77-89 years]; 1293 [82.9%] were female). After matching, overlapping hip fracture procedures had a greater risk for a complication (hazard ratio [HR], 1.85; 95% CI, 1.27-2.71; P = .001), as did overlapping THA procedures (HR, 1.79; 95% CI, 1.02-3.14; P = .04). Among overlapping hip fracture operations, increasing duration of operative overlap was associated with increasing risk for complications (adjusted odds ratio, 1.07 per 10-minute increase in overlap; P = .009). Overlapping surgery was relatively rare but was associated with an increased risk for surgical complications. Furthermore, increasing duration of operative overlap was associated with an increasing risk for complications. These findings support the notion that overlapping provision of surgery should be part of the informed consent process.


The evolution, diversity, and host associations of rhabdoviruses.
Metagenomic studies are leading to the discovery of a hidden diversity of RNA viruses. These new viruses are poorly characterized and new approaches are needed predict the host species these viruses pose a risk to. The rhabdoviruses are a diverse family of RNA viruses that includes important pathogens of humans, animals, and plants. We have discovered thirty-two new rhabdoviruses through a combination of our own RNA sequencing of insects and searching public sequence databases. Combining these with previously known sequences we reconstructed the phylogeny of 195 rhabdovirus sequences, and produced the most in depth analysis of the family to date. In most cases we know nothing about the biology of the viruses beyond the host they were identified from, but our dataset provides a powerful phylogenetic approach to predict which are vector-borne viruses and which are specific to vertebrates or arthropods. By reconstructing ancestral and present host states we found that switches between major groups of hosts have occurred rarely during rhabdovirus evolution. This allowed us to propose seventy-six new likely vector-borne vertebrate viruses among viruses identified from vertebrates or biting insects. Based on currently available data, our analysis suggests it is likely there was a single origin of the known plant viruses and arthropod-borne vertebrate viruses, while vertebrate- and arthropod-specific viruses arose at least twice. There are also few transitions between aquatic and terrestrial ecosystems. Viruses also cluster together at a finer scale, with closely related viruses tending to be found in closely related hosts. Our data therefore suggest that throughout their evolution, rhabdoviruses have occasionally jumped between distantly related host species before spreading through related hosts in the same environment. This approach offers a way to predict the most probable biology and key traits of newly discovered viruses.


Effects of an elemental diet, inert bulk and different types of dietary fibre on the response of the intestinal epithelium to refeeding in the rat and relationship to plasma gastrin, enteroglucagon, and PYY concentrations.
Refeeding starved rats with an elemental diet resulted in a marked increase in crypt cell production rate (CCPR) in the proximal small intestine but not in the distal regions of the gut. Little effect on CCPR was noted when inert bulk (kaolin) was added to the elemental diet. Addition of a poorly fermentable dietary fibre (purified wood cellulose) had little effect on intestinal epithelial cell proliferation except in the distal colon where it significantly increased CCPR. A more readily fermentable fibre (purified wheat bran) caused a large proliferative response in the proximal, mid, and distal colon and in the distal small intestine. A gel forming fibre only significantly stimulated proliferation in the distal colon; the rats in this group, however, did not eat all the food given. There was no significant correlation between CCPR and plasma gastrin concentrations, but plasma enteroglucagon concentrations were significantly correlated with CCPR in almost all the sites studied. Plasma PYY concentrations also showed some correlation with CCPR, especially in the colon. Thus while inert bulk cannot stimulate colonic epithelial cell proliferation fermentable fibre is capable of stimulating proliferation in the colon, and especially in the distal colon: it can also stimulate proliferation in the distal small intestine and it is likely that plasma enteroglucagon may have a role to play in this process.


Anti-Donor Immune Responses Elicited by Allogeneic Mesenchymal Stem Cells and Their Extracellular Vesicles: Are We Still Learning?
Mesenchymal stromal cells (MSC) have been used to treat a broad range of disease indications such as acute and chronic inflammatory disorders, autoimmune diseases, and transplant rejection due to their potent immunosuppressive/anti-inflammatory properties. The breadth of their usage is due in no small part to the vast quantity of published studies showing their ability to modulate multiple immune cell types of both the innate and adaptive immune response. While patient-derived (autologous) MSC may be the safer choice in terms of avoiding unwanted immune responses, factors including donor comorbidities may preclude these cells from use. In these situations, allogeneic MSC derived from genetically unrelated individuals must be used. While allogeneic MSC were initially believed to be immune-privileged, substantial evidence now exists to prove otherwise with multiple studies documenting specific cellular and humoral immune responses against donor antigens following administration of these cells. In this article, we will review recent published studies using non-manipulated, inflammatory molecule-activated (licensed) and differentiated allogeneic MSC, as well as MSC extracellular vesicles focusing on the immune responses to these cells and whether or not such responses have an impact on allogeneic MSC-mediated safety and efficacy.


Gamma delta T-cell neoplasms: a clinicopathological study of 11 cases.
The majority of T-cell neoplasms express T-cell antigen receptor (TCR) alpha beta on their cell surface, and a few cases show the TCR gamma delta phenotype. Recently, a variety of gamma delta T-cell neoplasm was recognized; however, its clinicopathological features have not been extensively analyzed. Here we report the results of a clinicopathological study of 11 cases of gamma delta T-cell neoplasm. During the 11-year period from 1989 to 1999, 104 patients with T-cell neoplasms were examined by flow cytometric analysis and/or immunohistochemical analysis. Tumor cells from all 104 patients expressed one or more of the T-cell antigens-CD2, CD3, CD5 and CD7. Forty-nine of the 104 cases of T-cell neoplasms were examined immunophenotypically for TCR alpha beta/gamma delta subsets. Expression of TCR gamma delta on tumor cells was found in five (33%) of 15 patients with precursor T-cell lymphoblastic leukemia/lymphoma, one (25%) of four with T-cell granular lymphocytic leukemia and five (26%) of 19 with peripheral T-cell lymphoma (PTCL), whereas no expression was found in 11 patients with adult T-cell leukemia-lymphoma. Primary sites of the five patients with gamma delta PTCL were as follows: lymph node, three; skin, one and liver, tonsil and skin, one. The courses of the three patients with gamma delta PTCL of nodal onset were very short (3, 5 and 9 months, respectively), and they were all resistant to combination chemotherapies. Although gamma delta T-cell neoplasm constitutes a heterogeneous population, it is important to examine the expression of TCR with the view to identifying possible poor prognostic subgroups, such as primary nodal gamma delta T-cell lymphoma.


Reasons for non-adherence to cardiometabolic medications, and acceptability of an interactive voice response intervention in patients with hypertension and type 2 diabetes in primary care: a qualitative study.
This study explored the reasons for patients' non-adherence to cardiometabolic medications, and tested the acceptability of the interactive voice response (IVR) as a way to address these reasons, and support patients, between primary care consultations. The study included face-to-face interviews with 19 patients with hypertension and/or type 2 diabetes mellitus, selected from primary care databases, and presumed to be non-adherent. Thirteen of these patients pretested elements of the IVR intervention few months later, using a think-aloud protocol. Five practice nurses were interviewed. Data were analysed using multiperspective, and longitudinalthematic analysis. Negative beliefs about taking medications, the complexity of prescribed medication regimens, and the limited ability to cope with the underlying affective state, within challenging contexts, were mentioned as important reasons for non-adherence. Nurses reported time constraints to address each patient's different reasons for non-adherence, and limited efficacy to support patients, between primary care consultations. Patients gave positive experiential feedback about the IVR messages as a way to support them take their medicines, and provided recommendations for intervention content and delivery mode. Specifically, they liked the voice delivering the messages and the voice recognition software. For intervention content, they preferred messages that were tailored, and included messages with 'information about health consequences', 'action plans', or simple reminders for performing the behaviour. Patients with hypertension and/or type 2 diabetes, and practice nurses, suggested messages tailored to each patient's reasons for non-adherence. Participants recommended IVR as an acceptable platform to support adherence to cardiometabolic medications between primary care consultations. Future studies could usefully test the acceptability, and feasibility, of tailored IVR interventions to support medication adherence, as an adjunct to primary care.


Two-Dimensional Nucleation on the Terrace of Colloidal Crystals with Added Polymers.
Understanding nucleation dynamics is important both fundamentally and technologically in materials science and other scientific fields. Two-dimensional (2D) nucleation is the predominant growth mechanism in colloidal crystallization, in which the particle interaction is attractive, and has recently been regarded as a promising method to fabricate varieties of complex nanostructures possessing innovative functionality. Here, polymers are added to a colloidal suspension to generate a depletion attractive force, and the detailed 2D nucleation process on the terrace of the colloidal crystals is investigated. In the system, we first measured the nucleation rate at various area fractions of particles on the terrace, ϕarea. In situ observations at single-particle resolution revealed that nucleation behavior follows the framework of classical nucleation theory (CNT), such as single-step nucleation pathway and existence of critical size. Characteristic nucleation behavior is observed in that the nucleation and growth stage are clearly differentiated. When many nuclei form in a small area of the terrace, a high density of kink sites of once formed islands makes growth more likely to occur than further nucleation because nucleation has a higher energy barrier than growth. The steady-state homogeneous 2D nucleation rate, J, and the critical size of nuclei, r*, are measured by in situ observations based on the CNT, which enable us to obtain the step free energy, γ, which is an important parameter for characterizing the nucleation process. The γ value is found to change according to the strength of attraction, which is tuned by the concentration of the polymer as a depletant.


Electrosensory Contrast Signals for Interacting Weakly Electric Fish.
Active sensory systems have evolved to properly encode natural stimuli including those created by conspecifics, yet little is known about the properties of such stimuli. We consider the electrosensory signal at the skin of a fixed weakly electric fish in the presence of a swimming conspecific. The dipole recordings are obtained in parallel with video tracking of the position of the animals. This enables the quantification of the relationships between the recording dipole and the positions of the head, midbody and tail of the freely swimming fish. The contrast of the signal at the skin is shown to be well-fitted by a decreasing exponential function of distance. It is thus anti-correlated with distance; it is also correlated with the second envelope (i.e., the envelope of the envelope) of the raw recorded signal. The variance of the contrast signal is highest at short range. However, the coefficient of variation (CV) of this signal increases with distance. We find a range of position and associated contrast patterns under quasi-2D swimming conditions. This is quantified using global measures of the visit times of the free fish within measurable range, with each visit causing a bump in contrast. The durations of these bumps as well as the times between these bumps are well reproduced by a doubly stochastic process formed by a dichotomous (two-state) noise with Poisson statistics multiplying a colored noise [Ornstein-Uhlenbeck (OU) process]. Certain rapid body movements such as bending or turning are seen to produce contrast drops that may be part of cloaking strategies.


Flexible theta sequence compression mediated via phase precessing interneurons.
Encoding of behavioral episodes as spike sequences during hippocampal theta oscillations provides a neural substrate for computations on events extended across time and space. However, the mechanisms underlying the numerous and diverse experimentally observed properties of theta sequences remain poorly understood. Here we account for theta sequences using a novel model constrained by the septo-hippocampal circuitry. We show that when spontaneously active interneurons integrate spatial signals and theta frequency pacemaker inputs, they generate phase precessing action potentials that can coordinate theta sequences in place cell populations. We reveal novel constraints on sequence generation, predict cellular properties and neural dynamics that characterize sequence compression, identify circuit organization principles for high capacity sequential representation, and show that theta sequences can be used as substrates for association of conditioned stimuli with recent and upcoming events. Our results suggest mechanisms for flexible sequence compression that are suited to associative learning across an animal's lifespan.


Precise, real-time, every-single-shot, carrier-envelope phase measurement of ultrashort laser pulses.
In this Letter we demonstrate a method for real-time determination of the carrier-envelope phase of each and every single ultrashort laser pulse at kilohertz repetition rates. The technique expands upon the recent work of Wittmann and incorporates a stereographic above-threshold laser-induced ionization measurement and electronics optimized to produce a signal corresponding to the carrier-envelope phase within microseconds of the laser interaction, thereby facilitating data-tagging and feedback applications. We achieve a precision of 113 mrad (6.5°) over the entire 2π range.


Trabecular architecture can remain intact for both disuse and overload enhanced resorption characteristics.
The paradigm that bone metabolic processes are controlled by osteocyte signals have been the subject of investigation in many recent studies. One hypothesis is that osteoblast formation is enhanced by these signals, and that osteoclast resorption is enhanced by the lack of them. Reduced, or absent, osteocyte signaling can be an effect of reduced mechanical loading (disuse) or of defects in the canalicular network, due to microcracks. This would mean that bone is resorbed precisely there where it is mostly needed. In our study, we addressed this apparent contradiction. The purpose was to investigate how alternative strain-based local stimuli for osteoclasts to resorb bone would affect remodeling and adaptation of the trabecular architecture. For this purpose, a computer-simulation model was used, which couples morphological and mechanical effects of local bone metabolism to changes in trabecular architecture and density at large. Six resorption characteristics were studied in the model: (I) resorption occurs spatially random, (II) resorption is enhanced or (III) strongly enhanced where there is disuse, (IV) resorption is enhanced or (V) strongly enhanced where there are high strains, i.e. overload, and (VI) resorption is enhanced where there is disuse and where there are high strains. Results showed that the rates of structural adaptation to alternative loading were higher for disuse-controlled resorption than for overload-controlled resorption. Architecture and mass remained stable for all cases except (V) in which the structure deteriorated as in osteoporotic bone. We conclude that, given the potential of osteoblasts to form bone in highly strained areas, based on signals from osteocytes, osteoclast resorption can normally be compensated for.


Survivin as a prognostic/predictive marker and molecular target in cancer therapy.
Evasion from apoptotic cell death is reported to be a pivotal mechanism by which tumor cells acquire resistance to therapeutic treatment. Targeting the apoptotic pathways may constitute a promising strategy to counteract therapy resistance and to re-sensitize cancer cells. Expression of survivin, the smallest and structurally unique member of the inhibitor of apoptosis protein (IAP) family, has been shown to be associated with poor clinical outcome, more aggressive clinicopathologic features and resistance to both, conventional chemo and radiation therapy. Moreover, survivin detection in cancer tissue, in circulating tumor cells and in patient's serum has prognostic and predictive relevance and may display a prerequisite for marker based molecular therapies. Indeed, due to its universal over expression in malignant tissue, and its prominent role at disparate networks of cellular division, intracellular signaling, apoptosis and adaption to unfavorable surroundings, survivin has been shown to be a suitable target for a targeted therapy. The applicability of survivindriven strategies in clinical practice is currently under investigation as the first survivin antagonists (small molecule inhibitors, antisense oligonucleotides and immunotherapy) successfully entered phase I/II trials. Taken together, these data provide a rationale for the implementation of both, survivin as a molecular diagnostic tool and survivin targeted therapies, within future clinical practice.


NLRP10 is a NOD-like receptor essential to initiate adaptive immunity by dendritic cells.
NLRs (nucleotide-binding domain leucine-rich-repeat-containing receptors; NOD-like receptors) are a class of pattern recognition receptor (PRR) that respond to host perturbation from either infectious agents or cellular stress. The function of most NLR family members has not been characterized and their role in instructing adaptive immune responses remains unclear. NLRP10 (also known as PYNOD, NALP10, PAN5 and NOD8) is the only NLR lacking the putative ligand-binding leucine-rich-repeat domain, and has been postulated to be a negative regulator of other NLR members, including NLRP3 (refs 4-6). We did not find evidence that NLRP10 functions through an inflammasome to regulate caspase-1 activity nor that it regulates other inflammasomes. Instead, Nlrp10(-/-) mice had a profound defect in helper T-cell-driven immune responses to a diverse array of adjuvants, including lipopolysaccharide, aluminium hydroxide and complete Freund's adjuvant. Adaptive immunity was impaired in the absence of NLRP10 because of a dendritic cell (DC) intrinsic defect in emigration from inflamed tissues, whereas upregulation of DC costimulatory molecules and chemotaxis to CCR7-dependent and -independent ligands remained intact. The loss of antigen transport to the draining lymph nodes by a subset of migratory DCs resulted in an almost absolute loss in naive CD4(+) T-cell priming, highlighting the critical link between diverse innate immune stimulation, NLRP10 activity and the immune function of mature DCs.


Molecular profiling of a lethal tumor microenvironment, as defined by stromal caveolin-1 status in breast cancers.
Breast cancer progression and metastasis are driven by complex and reciprocal interactions, between epithelial cancer cells and their surrounding stromal microenvironment. We have previously shown that a loss of stromal Cav-1 expression is associated with an increased risk of early tumor recurrence, metastasis and decreased overall survival. To identify and characterize the signaling pathways that are activated in Cav-1 negative tumor stroma, we performed gene expression profiling using laser microdissected breast cancer-associated stroma. Tumor stroma was laser capture microdissected from 4 cases showing high stromal Cav-1 expression and 7 cases with loss of stromal Cav-1. Briefly, we identified 238 gene transcripts that were upregulated and 232 gene transcripts that were downregulated in the stroma of tumors showing a loss of Cav-1 expression (p ≤ 0.01 and fold-change ≥ 1.5). Gene set enrichment analysis (GSEA) revealed "stemness," inflammation, DNA damage, aging, oxidative stress, hypoxia, autophagy and mitochondrial dysfunction in the tumor stroma of patients lacking stromal Cav-1. Our findings are consistent with the recently proposed "Reverse Warburg Effect" and the "Autophagic Tumor Stroma Model of Cancer Metabolism." In these two complementary models, cancer cells induce oxidative stress in adjacent stromal cells, which then forces these stromal fibroblasts to undergo autophagy/mitophagy and aerobic glycolysis. This, in turn, produces recycled nutrients (lactate, ketones and glutamine) to feed anabolic cancer cells, which are undergoing oxidative mitochondrial metabolism. Our results are also consistent with previous biomarker studies showing that the increased expression of known autophagy markers (such as ATG16L and the cathepsins) in the tumor stroma is specifically associated with metastatic tumor progression and/or poor clinical outcome.


The Association Between Smartphone Use and Breast Cancer Risk Among Taiwanese Women: A Case-Control Study.
Breast cancer is a common malignancy worldwide. Smartphones have gradually become indispensable to our modern lives and have already changed lifestyles of human beings. To our best knowledge, no study has investigated the relationship between smartphone use and breast cancer. This case-control study purposely investigated the relationship between smartphone use and breast cancer risk. This was a case-control study comprising 894 healthy controls and 211 patients with breast cancer. All participants were asked to respond to standard questionnaires to collect information on sleep quality, smartphone addiction, and smartphone use. Participants with smartphone addiction had a significantly higher 1.43-fold risk of breast cancer. Individuals with the habitual behavior of smartphone use >4.5 minutes before bedtime had a significantly increased 5.27-fold risk of breast cancer compared to those who used a smartphone for ≤4.5 minutes before bedtime. Additionally, a closer distance between the smartphone and the breasts when using the smartphone exhibited a significantly increased 1.59-fold risk. Participants who carried their smartphone near their chest or waist-abdomen area had significantly increased 5.03-fold and 4.06-fold risks of breast cancer, respectively, compared to those who carried the smartphone below the waist. Moreover, there was a synergistic effect of smartphone addiction and smartphone use of >4.5 minutes before bedtime which increased the breast cancer risk. Excessive smartphone use significantly increased the risk of breast cancer, particularly for participants with smartphone addiction, a close distance between the breasts and smartphone, and the habit of smartphone use before bedtime.


A VLSI Systolic Architecture for Pattern Clustering.
Cluster analysis is a valuable tool in exploratory pattern analysis, especially when very little prior information about the data is available. In unsupervised pattern recognition and image segmentation applications, clustering techniques play an important role. The squared-error clustering technique is the most popular one among different clustering techniques. Due to the iterative nature of the squared-error clustering, it demands substantial CPU time, even for modest numbers of patterns. Recent advances in VLSI microelectronic technology triggered the idea of implementing the squared-error clustering directly in hardware. A two-level pipelined systolic pattern clustering array is proposed in this paper. The memory storage and access schemes are designed to enable a rhythmic data flow between processing units. Each processing unit is pipelined to further enhance the system performance. The total processing time for each pass of pattern labeling and cluster center updating is essentially dominated by the time required to fetch the pattern matrix once. Detailed architectural configuration, system performance evaluation, and simulation experiments are presented. The modularity and the regularity of the system architecture make it suited for VLSI implementations.


17O and 15N solid state NMR studies on ligand-assisted templating and oxygen coordination in the walls of mesoporous Nb, Ta and Ti oxides.
A multinuclear solid state NMR approach is applied to four templated mesoporous oxides (silica, titania, niobia and tantala) to include (15)N and (17)O magic angle spinning (MAS) NMR and double resonance (15)N-(93)Nb, (17)O Rotational-Echo Adiabatic Passage Double Resonance (REAPDOR). The templated samples were ramped in steps of 20 degrees C for 2 days up to typically 110 degrees C where the samples were left for 2-4 days. (15)N MAS NMR shows that amines are the only species present in the TiO2, Nb2O5, and Ta2O5. In SiO2, amines are only present as a minor coordination (10 +/- 2%), but there are several strong ammonium (15)N resonances. The REAPDOR experiments show that the nitrogen interacts with niobium, confirming a ligand interaction between the Nb and N, as previously believed. In the case of silica, the amine is quaternized and there is apparently no interaction with the Si, suggesting a RNH3(+) (-)O-Si- hydrogen-bonding interaction with the walls. (17)O MAS NMR provides the clearest indication of the local wall structure. In the aged, templated samples in all cases only OM2 coordinations are present which is very different from the pure bulk oxides (apart from SiO2) and must be due to the effects of amine coordination at the metal centers. On removal of the template, these oxides behave differently, with Ta2O5 showing a mixture of OTa2 (85 +/- 5%) and OTa3 (15 +/- 5%) which is similar to the types of coordination found in the bulk oxide. The previously reported (17)O MAS NMR data from heat-treated mesoporous niobia shows only ONb2, which is very highly ordered. In contrast for titania, the OTi2 coordination is immediately lost on removal of the template to be replaced by a mixture of OTi3 (60 +/- 5%) and OTi4 (40 +/- 5%), with the OTi4 becoming dominant above 250 degrees C, very different behavior from the corresponding bulk oxide. In summary, this NMR study shows that the local oxygen coordination in amine-templated mesoporous transition metal oxides is present as OM2 which is relatively rare in bulk oxides. The data indicates that the template interaction is largely controlled by the N-M dative bond to the wall, suppressing higher oxygen coordination numbers. Qualitatively it appears that the strength of this interaction varies greatly in the different mesoporous oxides.


Modeling gene and genome duplications in eukaryotes.
Recent analysis of complete eukaryotic genome sequences has revealed that gene duplication has been rampant. Moreover, next to a continuous mode of gene duplication, in many eukaryotic organisms the complete genome has been duplicated in their evolutionary past. Such large-scale gene duplication events have been associated with important evolutionary transitions or major leaps in development and adaptive radiations of species. Here, we present an evolutionary model that simulates the duplication dynamics of genes, considering genome-wide duplication events and a continuous mode of gene duplication. Modeling the evolution of the different functional categories of genes assesses the importance of different duplication events for gene families involved in specific functions or processes. By applying our model to the Arabidopsis genome, for which there is compelling evidence for three whole-genome duplications, we show that gene loss is strikingly different for large-scale and small-scale duplication events and highly biased toward certain functional classes. We provide evidence that some categories of genes were almost exclusively expanded through large-scale gene duplication events. In particular, we show that the three whole-genome duplications in Arabidopsis have been directly responsible for >90% of the increase in transcription factors, signal transducers, and developmental genes in the last 350 million years. Our evolutionary model is widely applicable and can be used to evaluate different assumptions regarding small- or large-scale gene duplication events in eukaryotic genomes.


Genomic identification of founding haplotypes reveals the history of the selfing species Capsella rubella.
The shift from outcrossing to self-fertilization is among the most common evolutionary transitions in flowering plants. Until recently, however, a genome-wide view of this transition has been obscured by both a dearth of appropriate data and the lack of appropriate population genomic methods to interpret such data. Here, we present a novel population genomic analysis detailing the origin of the selfing species, Capsella rubella, which recently split from its outcrossing sister, Capsella grandiflora. Due to the recency of the split, much of the variation within C. rubella is also found within C. grandiflora. We can therefore identify genomic regions where two C. rubella individuals have inherited the same or different segments of ancestral diversity (i.e. founding haplotypes) present in C. rubella's founder(s). Based on this analysis, we show that C. rubella was founded by multiple individuals drawn from a diverse ancestral population closely related to extant C. grandiflora, that drift and selection have rapidly homogenized most of this ancestral variation since C. rubella's founding, and that little novel variation has accumulated within this time. Despite the extensive loss of ancestral variation, the approximately 25% of the genome for which two C. rubella individuals have inherited different founding haplotypes makes up roughly 90% of the genetic variation between them. To extend these findings, we develop a coalescent model that utilizes the inferred frequency of founding haplotypes and variation within founding haplotypes to estimate that C. rubella was founded by a potentially large number of individuals between 50 and 100 kya, and has subsequently experienced a twenty-fold reduction in its effective population size. As population genomic data from an increasing number of outcrossing/selfing pairs are generated, analyses like the one developed here will facilitate a fine-scaled view of the evolutionary and demographic impact of the transition to self-fertilization.


George E. Brown memorial lecture. Oxygen radicals in cerebral vascular injury.
Acute, severe increases in arterial blood pressure cause sustained cerebral arteriolar dilation, abnormal reactivity to carbon dioxide and to changes in blood pressure, abolition of endothelium-dependent dilation from acetylcholine, discrete morphological lesions of the endothelium and vascular smooth muscle, and breakdown of the blood-brain barrier to plasma proteins. The dilation, abnormal reactivity, and morphological abnormalities are inhibited by pretreatment with cyclooxygenase inhibitors or with free radical scavengers. Superoxide dismutase-inhibitable reduction of nitroblue tetrazolium applied to the brain surface was detectable both during hypertension and one hour after hypertension subsided. Nitroblue tetrazolium reduction is also reduced by inhibitors of the anion channel. The abnormalities seen after hypertension are reproduced by topical application of arachidonate. The results are consistent with the view that acute hypertension induces generation of superoxide anion radical in association with accelerated arachidonate metabolism via cyclooxygenase. This radical enters cerebral extracellular space via the anion channel and gives rise to hydrogen peroxide and hydroxyl radical. All three radicals are capable of causing vasodilation by relaxation of cerebral vascular smooth muscle. The hydroxyl radical is the most likely candidate for vascular wall damage. The significance of this mechanism in chronic experimental hypertension or its relevance to human disease is not known.


WASH for WORMS: A Cluster-Randomized Controlled Trial of the Impact of a Community Integrated Water, Sanitation, and Hygiene and Deworming Intervention on Soil-Transmitted Helminth Infections.
Water, sanitation, and hygiene (WASH) interventions have been proposed as an important complement to deworming programs for sustainable control of soil-transmitted helminth (STH) infections. We aimed to determine whether a community-based WASH program had additional benefits in reducing STH infections compared with community deworming alone. We conducted the WASH for WORMS cluster-randomized controlled trial in 18 rural communities in Timor-Leste. Intervention communities received a WASH intervention that provided access to an improved water source, promoted improved household sanitation, and encouraged handwashing with soap. All eligible community members in intervention and control arms received albendazole every 6 months for 2 years. The primary outcomes were infection with each STH, measured using multiplex real-time quantitative polymerase chain reaction. We compared outcomes between study arms using generalized linear mixed models, accounting for clustering at community, household, and individual levels. At study completion, the integrated WASH and deworming intervention did not have an effect on infection with Ascaris spp. (relative risk [RR] 2.87, 95% confidence interval [CI]: 0.66-12.48, P = 0.159) or Necator americanus (RR 0.99, 95% CI: 0.52-1.89, P = 0.987), compared with deworming alone. At the last follow-up, open defecation was practiced by 66.1% (95% CI: 54.2-80.2) of respondents in the control arm versus 40.2% (95% CI: 25.3-52.6) of respondents in the intervention arm (P = 0.005). We found no evidence that the WASH intervention resulted in additional reductions in STH infections beyond that achieved with deworming alone over the 2-year trial period. The role of WASH on STH infections over a longer period of time and in the absence of deworming remains to be determined.


Food and oral health: 1. Dental caries.
Dental schools tend to teach only the rudiments of this subject and this does not provide a good basis for dentists to advise patients and combat the misinformation surrounding the topic. The first article in the two-part series examines nutrition as it applies to dental caries, while part two will discuss some of the general principles of nutrition in relation to oral health, periodontal disease and the oral mucosa.


Effect of paracetamol (acetaminophen) on body temperature in acute ischemic stroke: a double-blind, randomized phase II clinical trial.
Body temperature is a strong predictor of outcome in acute stroke. However, it is unknown whether antipyretic treatment leads to early and clinically worthwhile reduction of body temperature in patients with acute stroke, especially when they have no fever. The main purpose of this trial was to study whether early treatment of acute ischemic stroke patients with acetaminophen (paracetamol) reduces body temperature. Seventy-five patients with acute ischemic stroke confined to the anterior circulation were randomized to treatment with either 500 mg (low dose) or 1000 mg (high dose) acetaminophen or with placebo, administered as suppositories 6 times daily during 5 days. Body temperatures were measured with a rectal electronic thermometer at the start of treatment and after 24 hours and with an infrared tympanic thermometer at 2-hour intervals during the first 24 hours and at 6-hour intervals thereafter. The primary outcome measure was rectal temperature at 24 hours after the start of treatment. Treatment with high-dose acetaminophen resulted in 0.4 degrees C lower body temperatures than placebo treatment at 24 hours (95% CI 0.1 degrees C to 0.7 degrees C). The mean reduction from baseline temperature with high-dose acetaminophen was 0.3 degrees C (95% CI 0 degrees C to 0.6 degrees C) higher than that in placebo-treated patients. Treatment with low-dose acetaminophen did not result in lower body temperatures. After 5 days of treatment, no differences in temperature were found between the placebo and the high- or low-dose acetaminophen groups. Treatment with a daily dose of 6000 mg acetaminophen may result in a small, but potentially beneficial, decrease in body temperature shortly after ischemic stroke, even in normothermic and subfebrile patients. Further studies should determine whether this effect is reproducible and whether early reduction of body temperature leads to improved outcome.


E-learning for research capacity strengthening in sexual and reproductive health: the experience of the Geneva Foundation for Medical Education and Research and the Department of Reproductive Health and Research, World Health Organization.
Technological advancement has resulted in the increasing use of e-learning and online education, initially in high-income countries and increasingly in low- and middle-income countries. In 2010, the Geneva Foundation for Medical Education and Research, in collaboration with the World Health Organization and partner institutions, developed an online postgraduate course "From Research to Practice: Training Course in Sexual and Reproductive Health Research". This course takes advantage of the advancing Internet technology to provide training opportunities to health professionals mostly from low- and middle-income countries whose access to quality education is constrained by time, financial resources, or both. To assess the outcomes of the course, an evaluation was conducted by sending a self-administered questionnaire to graduates of the 2010-2012 programme. The objectives were to determine if the graduates had applied the knowledge gained from the course to their work and whether they had implemented their research project developed during the course. The evaluation also appraised the number of graduates who participated in the design or implementation of a new research project since the course concluded and whether the course had contributed to advancement in their careers. A total of 175 of 219 course graduates answered the questionnaire. The evaluation revealed that the majority of respondents (98%) had utilized the knowledge acquired, with nearly half of them (47%) having published a scientific paper as author or co-author. About a third of respondents (39%) had implemented their course research project and about three quarters of them (74%) have been involved in the design or implementation of a research project after completing the course. Over three quarters (81%) of respondents opined that the course had contributed to their career advancement and almost half of them (46%) had a career promotion as a direct or indirect benefit of the course. We surmise that the course positively impacted the participants' knowledge and understanding of sexual and reproductive health, which they applied in their professional work, as well as strengthened their research capacity. Success factors for the e-learning programme include tailor-made content to meet participants' needs, flexibility of access, and ongoing engagement/personal interactivity with course coaches.


Recursive estimation of 3D motion and surface structure from local affine flow parameters.
A recursive structure from motion algorithm based on optical flow measurements taken from an image sequence is described. It provides estimates of surface normals in addition to 3D motion and depth. The measurements are affine motion parameters which approximate the local flow fields associated with near-planar surface patches in the scene. These are integrated over time to give estimates of the 3D parameters using an extended Kalman filter. This also estimates the camera focal length and, so, the 3D estimates are metric. The use of parametric measurements means that the algorithm is computationally less demanding than previous optical flow approaches and the recursive filter builds in a degree of noise robustness. Results of experiments on synthetic and real image sequences demonstrate that the algorithm performs well.


Evolutionary conservation of the structure and expression of alternatively spliced Ultrabithorax isoforms from Drosophila.
In Drosophila melanogaster, alternatively spliced mRNAs from the homeotic gene Ultrabithorax (Ubx) encode a family of structurally distinct homeoprotein isoforms. The developmentally regulated expression patterns of these isoforms suggest that they have specialized stage- and tissue-specific functions. To evaluate the functional importance of UBX isoform diversity and gain clues to the mechanism that regulates processing of Ubx RNAs, we have investigated whether the Ubx RNAs of other insects undergo similar alternative splicing. We have isolated and characterized Ubx cDNA fragments from D. melanogaster, Drosophila pseudoobscura, Drosophila hydei and Drosophila virilis, species separated by as much as 60 million years of evolution, and have found that three aspects of Ubx RNA processing have been conserved. (1) These four species exhibit identical patterns of optional exon use in a region adjacent to the homeodomain. (2) These four species produce the same family of UBX protein isoforms with identical amino acid sequences in the optional exons, even though the common amino-proximal region has undergone substantial divergence. The nucleotide sequences of the optional exons, including third positions of rare codons, have also been conserved strongly, suggesting functional constraints that are not limited to coding potential. (3) The tissue- and stage-specific patterns of expression of different UBX isoforms are identical among these Drosophila species, indicating that the developmental regulation of the alternative splicing events has also been conserved. These findings argue for an important role of alternative splicing in Ubx function. We discuss the implications of these results for models of UBX protein function and the mechanism of alternative splicing.


Fossil Homo femur from Berg Aukas, northern Namibia.
The proximal half of a hominid femur was recovered from deep within a paleokarst feature at the Berg Aukas mine, northern Namibia. The femur is fully mineralized, but it is not possible to place it in geochronological context. It has a very large head, an exceptionally thick diaphyseal cortex, and a very low collodiaphyseal angle, which serve to differentiate it from Holocene homologues. The femur is not attributable to Australopithecus, Paranthropus, or early Homo (i.e., H. habilis sensu lato). Homo erectus femora have a relatively longer and AP flatter neck, and a shaft that exhibits less pilaster than the Berg Aukas specimen. Berg Aukas also differs from early modern femora in several features, including diaphyseal cortical thickness and the degree of subtrochanteric AP flattening. The massive diaphyseal cortex of Berg Aukas finds its closest similarity within archaic H. sapiens (e.g., Castel di Guido) and H. erectus (e.g., KNM-ER 736) samples. It has more cortical bone at midshaft than any other specimen, although relative cortical thickness and the asymmetry of its cross-sectional disposition at this level are comparable with those of other Pleistocene femora. The closest morphological comparisons with Berg Aukas are in archaic (i.e., Middle Pleistocene) H. sapiens and Neandertal samples.


Dissociation between pain and the nociceptive blink reflex during psychological arousal.
To investigate the effect of psychological arousal on pain ratings and the R2 component of the electrically evoked blink reflex to a 'pure' noiciceptive stimulus. Pain ratings and R2 to a noiciceptive stimulus (pulse width 0.3ms, 2mA, delivered from a concentric electrode attached to the supraorbital region of the forehead) were investigated in 16 healthy participants before and during a serial subtraction task, and in 16 control participants who sat quietly during nociceptive stimulation. Pain ratings decreased whereas R2 amplitude increased during the serial subtraction task. Supra-spinal rather than spinal mechanisms inhibited pain perception during psychological arousal. Moreover, psychological arousal facilitated the R2 component of the blink reflex to a nociception-specific stimulus. Supra-spinal influences need to be considered during clinical evaluation of the trigeminal nociceptive blink reflex.


Quantitative analysis of ultrasound B-mode images of carotid atherosclerotic plaque: correlation with visual classification and histological examination.
This paper presents a quantitative comparison of three types of information available for 52 patients scheduled for carotid endarterectomy: subjective classification of the ultrasound images obtained during scanning before operation, first-and second-order statistical features extracted from regions of the plaque in still ultrasound images from three orthogonal scan planes and finally a histological analysis of the surgically removed plaque. The quantitative comparison was made with the linear model and with separation of the available data into training and test sets. The comparison of subjective classification with features from still ultrasound images revealed an overall agreement of 60% for classification of echogenicity and 70% for classification of structure. Comparison of the histologically determined relative volume of soft materials with features from the still images revealed a correlation coefficient of r = -0.42(p = 0.002). for mean echogenicity of the plaque region. The best performing feature was of second order and denoted Contrast (r = -0.5). Though significant, the latter correlation is probably not strong enough to be useful for clinical prediction of relative volume of soft materials for individual patients. Reasons for this is discussed in the paper, together with suggestions for improvements.