Role of tumor-derived cytokines on the immune system of mice bearing a mammary adenocarcinoma. II. Down-regulation of macrophage-mediated cytotoxicity by tumor-derived granulocyte-macrophage colony-stimulating factor.
Peritoneal elicited macrophages (PEM) from mammary tumor-bearing mice have a decreased capacity to become cytotoxic against syngeneic, allogeneic, and xenogeneic target cells upon in vitro stimulation with LPS, as compared with PEM of normal mice. A regulatory mechanism other than PG release is suggested because the addition of both indomethacin and LPS to macrophage cultures from tumor-bearing mice caused no changes in their cytotoxic capability. Because tumor products have been implicated in the down-regulation of immune responses, we investigated whether pretreatment with supernatants from the tumor cell line DA-3, derived from the in vivo mammary adenocarcinoma D1-DMBA-3, affects the cytolytic capacity of macrophages. This treatment inhibits, in a dose-dependent fashion, the ability of stimulated normal PEM to kill target cells. Partial purification of DA-3 cell line supernatant showed that most of the inhibitory activity was exerted by factors with a molecular mass greater than 10 kDa and less than 30 kDa. However, slight inhibition could also be observed with fractions containing molecules less than 10 kDa. The data suggest that more than one factor released by the mammary tumor cells may be involved in the down-regulation of macrophage-mediated cytotoxicity. Because the DA-3 cells constitutively produce granulocyte-macrophage CSF (GM-CSF), which has a molecular mass of 27 kDa, we pretreated PEM from normal mice in vitro with rGM-CSF for 24 h. This resulted in a dose-dependent decrease in their capacity to kill tumor target cells upon LPS stimulation. Furthermore, PEM from normal mice injected with rGM-CSF for 25 days displayed a profound decrease in their cytolytic ability against DA-3 targets upon in vitro stimulation with increasing amounts of LPS. The pretreatment of PEM from normal mice with a combination of DA-3 cell supernatants and specific anti-GM-CSF partially neutralized the inhibitory effect of the DA-3 supernatant on macrophage tumoricidal capability. These results indicate that tumor-derived GM-CSF is an important factor involved in the decreased macrophage cytotoxicity during mammary adenocarcinoma progression.


Initial evaluation of a direct detection device detector for single particle cryo-electron microscopy.
We report on initial results of using a new direct detection device (DDD) for single particle reconstruction of vitreous ice embedded specimens. Images were acquired on a Tecnai F20 at 200keV and a nominal magnification of 29,000×. This camera has a significantly improved signal to noise ratio and modulation transfer function (MTF) at 200keV compared to a standard CCD camera installed on the same microscope. Control of the DDD has been integrated into Leginon, an automated data collection system. Using GroEL as a test specimen, we obtained images of ∼30K particles with the CCD and the DDD from the same specimen sample using essentially identical imaging conditions. Comparison of the maps reconstructed from the CCD images and the DDD images demonstrates the improved performance of the DDD. We also obtained a 3D reconstruction from ∼70K GroEL particles acquired using the DDD; the quality of the density map demonstrates the potential of this new recording device for cryoEM data acquisition.


HLA class II induction in human islet cells by interferon-gamma plus tumour necrosis factor or lymphotoxin.
HLA class II molecules are surface glycoproteins which are essential in the initiation of immune responses. It has been postulated that induction of class II in epithelial cells such as endocrine cells, which are normally class II negative, may result in autoimmunity. In type I diabetes, islet beta cells, the target of the autoimmune process, selectively express class II antigens. But in contrast to most other cell types, islet beta cells are not stimulated to express class II by interferon-gamma (IFN-gamma) and thus the conditions under which this induction occurs have been particularly elusive. The cytotoxins tumour necrosis factor (TNF) and lymphotoxin (LT) synergize with IFN-gamma in a number of activities. We report here that IFN-gamma in combination with either TNF or LT induces islet cell class II expression. This finding has important implications for the pathogenesis of type I diabetes and the understanding of the differential control of class II expression.


Impact of combined skin sparing mastectomy and immediate subpectoral prosthetic reconstruction on the pectoralis major muscle function: a preoperative and postoperative comparative study.
We assessed the subjective and objective functional loss induced by detachment of the pectoralis major muscle and whether synergistic function of adjacent muscles compensates for such loss. A preoperative to postoperative comparison of the Disability of the Arm, Shoulder and Hand score, 3-dimensional video-analysis of the range of motion, and dynamometric force assessment with electromyography registration were prospectively conducted in 22 women who underwent bilateral preventive breast surgery. Although the statistically significant postoperative increase of the Disability of the Arm, Shoulder and Hand score (P = < 0.01) was indicative of more subjective disability, this difference is irrelevant for activities of daily life. This is in line with the observed postoperative lack of restrictions of range of motion of the shoulder. The mean postoperative maximum force measured in 4 directions reflective of the pectoralis major muscle function was 7.13 N, or 3.8%, lower than the preoperative force (P = 0.079). The mean postoperative maximum force in the 2 contramovement directions was 15.5 N, or 14%, higher than the preoperative force (P = 0.04). Electromyographically, both parts of the deltoid muscle preoperatively and postoperatively showed the same peaks in the 6 directions. Contrastingly, the clavicular part of the pectoralis major muscle postoperatively showed a statistically significant increase of peak of muscle activity in 3 of 4 directions reflective of pectoralis major function and in both contramovement directions, indicating compensatory activity. Daily life consequences of partial detachment of the pectoralis major muscle were acceptable in our patients.


Amitriptyline for the treatment of fibromyalgia: a comprehensive review.
Fibromyalgia is characterized by chronic generalized pain accompanied by a wide range of clinical manifestations. Most clinical practice guidelines recommend multidisciplinary treatment using a combination of pharmacological and non-pharmacological therapies. The tricyclic antidepressant amitriptyline has been most thoroughly studied in fibromyalgia. Amitriptyline has been evaluated in placebo-controlled studies, and it has served as an active comparator to other therapeutic interventions in the treatment of fibromyalgia. In addition, several systematic reviews and meta-analyses have evaluated its efficacy and safety for the treatment of fibromyalgia. Data from individual studies as well as from systematic reviews indicate that low doses (10-75 mg/day) of amitriptyline are effective for the treatment of fibromyalgia and, despite the limited quality of the data, they do not seem to be associated with relevant tolerability or safety issues. Consistent with some clinical guidelines, we believe amitriptyline in low doses should be considered a first-line drug for the treatment of fibromyalgia.


Population awareness and control of hypertension and hypercholesterolemia. The Atherosclerosis Risk in Communities study.
A national program for hypertension detection and control was implemented in the 1970s, whereas a similar program for control of hypercholesterolemia has been implemented in recent years. We studied the levels of awareness, treatment, and control of these conditions in US population samples during a 3-year period (1987 to 1989). The levels of awareness, treatment (by medication), and adequate control of hypertension (systolic blood pressure, > or = 140 mm Hg; diastolic blood pressure, > or = 90 mm Hg; or antihypertensive medication) and hypercholesterolemia (serum cholesterol level, > or = 6.21 mmol/L [> or = 240 mg/dL], or lipid-lowering medication) were studied among participants in the baseline examination of the Atherosclerosis Risk in Communities Study, including 15,739 individuals aged 45 to 64 years. Eighty-four percent of the hypertensive subjects and 42% of the hypercholesterolemic subjects were aware of their conditions. Overall, 50% of the hypertensive subjects and only 4% of the hypercholesterolemic subjects had their conditions both treated and controlled. Rates of hypertension prevalence, awareness, and control remained stable during the 3-year study period. Hypercholesterolemia prevalence decreased from 30% in 1987 to 25% in 1989; its awareness increased from 31% to 50% during the same period. Hypertensive women were more likely than hypertensive men to be aware and treated, whereas hypercholesterolemia awareness was higher in men than in women. Hypertension awareness was highest in black women, but black hypertensive subjects were less likely than whites to be treated and to have their hypertension controlled. Black hypercholesterolemic subjects were less likely to be either aware or treated. After the recent implementation of the National Cholesterol Education Program, the levels of awareness, treatment, and control of hypercholesterolemia are improving at a high rate, although they are still substantially lower than those for hypertension. Further improvement is necessary, particularly among certain population groups, such as blacks.


A prospective study of emotional disorders in childbearing women.
A group of first-time mothers (119) were interviewed repeatedly at fixed intervals during their pregnancies and until their babies were a year old; they were then followed up at four years. A similar investigation was carried out on 38 other primiparae and 39 multiparae, but only postnatally. The incidence of depressive neurosis rose significantly in early pregnancy and in the first three months after delivery (10 per cent and 14 per cent of the main sample respectively). Subjects mainly suffered either from antenatal or postnatal depression, not both. Marital conflict and severe doubts about having the baby were associated with depression at either time. Bereavement and preterm birth were the only life events to relate with the onset of depression and bereavement had a greater impact during pregnancy. Depressed mothers were more likely to express negative or mixed feelings about their three-month-old babies. Many who had become depressed for the first time in their lives continued to experience psychological problems for up to four years after childbirth.


A polysaccharide from Trametes robiniophila Murrill induces apoptosis through intrinsic mitochondrial pathway in human osteosarcoma (U-2 OS) cells.
In this study, we isolated and purified one homogeneous polysaccharide (TRP) from the fruiting bodies of Trametes robiniophila Murrill, and its average molecular weight was estimated to be 8.7 × 10(4) Da. Monosaccharide composition analysis by gas chromatography (GC) indicated that TRP was composed of glucose, galactose, and arabinose in the molar ratio of 4.2:1.10:1.06. Particularly, we evaluated the anti-cancer efficacy of TRP on human osteosarcoma U-2 OS cells in vitro and associated possible molecular mechanism. Our result provided the first evidence that treatment of U-2 OS cells with TRP resulted in a dose- and time-dependent inhibitory effect on cell proliferation of U-2 OS cells and caused apoptotic death. Moreover, TRP induced the apoptosis of U-2 OS cells via a mitochondria-dependent pathway, as evidenced by an increase in Bax/Bcl-2 ratio, a loss of mitochondrial membrane potential (Δψm), release of cytochrome c from the mitochondria to the cytosol, activation of caspase-9 and caspase-3, and cleavage of poly(ADP-ribose) polymerase (PARP) in U-2 OS cells. In addition, overexpression of metadherin (MTDH), one carcinogene, was inhibited in U-2 OS cells after exposure to TRP for 24 h. Our findings suggested that TRP inhibited the proliferation of human osteosarcoma cancer cells by promoting apoptosis through the intrinsic mitochondrial pathway, as well as inhibition of MTDH expression.


Accuracy of reporting bleeding during pregnancy.
Vaginal bleeding during pregnancy has been considered a marker of an at-risk pregnancy, but the accuracy of reported bleeding has not been assessed. We sought to evaluate the agreement in vaginal bleeding reports based on prospective daily diary and retrospective recall at first-trimester interview and to investigate predictors of reporting accuracy. Participants recruited prior to pregnancy for a community-based pregnancy cohort (n = 153) completed web-based daily diaries beginning before pregnancy until the end of the first trimester. A comprehensive first-trimester interview was conducted, and the bleeding data from diary and interview were compared. Kappa statistics were used to quantify agreement. Log-linear models were used to investigate maternal age, prior miscarriage, and current pregnancy outcome as potential predictors of agreement. We found that bleeding characteristics (number of bleeding episodes, bleeding heaviness, duration and gestational timing) from the diary and interview were reported with high levels of agreement. Kappas ranged from 0.77 to 0.84. Retrospective report of any bleeding had a sensitivity of 0.80 and specificity of 1.0; however, sensitivity was lower when examined within smaller time intervals. Important predictors of agreement were not identified in this analysis, but the sample was small. Overall, the presence of vaginal bleeding, a common and potentially alarming symptom of early pregnancy, may be assessed by interview later in pregnancy with reasonable accuracy.


Long-term follow-up of a prospective study of combined modality therapy for stage I-II indolent non-Hodgkin's lymphoma.
Standard therapy for patients with stage I-II indolent lymphoma has been involved-field radiation therapy (IF-XRT), which achieves 10-year disease-free survival in 40% to 50% of patients, with many of these patients cured. We investigated the potential for combined-modality therapy to increase the disease-free survival for such patients. A total of 102 eligible patients with stage I-II low grade lymphoma (International Working Formulation criteria) were enrolled from 1984 to 1992. Treatment comprised 10 cycles of risk-adapted chemotherapy (cyclophosphamide, vincristine, prednisone, bleomycin [COP-Bleo], and with doxorubicin added for some [CHOP-Bleo]) and 30 to 40 Gy IF-XRT. The patients' median age was 56 years (range, 28 to 77), with follicular histology in 83%, bulky disease (>/= 5 cm) in 24%, and stage II in 52%. There were no treatment-related deaths and 99% of patients attained complete remission. With a median follow-up of 10 years, the 10-year time to treatment failure and overall survival were 76% and 82%, respectively. For patients with follicular lymphoma, these figures were 72% and 80%, respectively. The only factor associated with treatment failure, for follicular lymphoma patients, was stage-modified International Prognostic Factors Index score (P =.02). None of 17 patients with diffuse small lymphocytic or mucosa-associated lymphoid tissue histology have relapsed. Elevated serum beta2-microglobulin was associated with shorter survival (P <.0001). The 10-year survival after relapse was 46%. There have been two cases of myelodysplasia and 12 other new malignancies, including four arising within radiation fields. With prolonged follow-up, combined-modality therapy with risk-adapted COP-/CHOP-Bleo and IF radiation has attained higher rates of disease control and survival than previously reported with IF-XRT alone. This apparent improvement is being further explored in an ongoing randomized trial.


Density-related effects on the infectivity and aggressiveness of a sterilising smut in a wild population of Digitaria sanguinalis.
Understanding host-pathogen evolutionary dynamics needs characterisation and quantification of processes occurring at many spatiotemporal scales. With this aim, the effects of smut on a naturally infected population of the summer annual Digitaria sanguinalis were followed for 4 years in an uncropped field. The main purpose of the study was to quantify the effects of within-population density on the infectivity and the aggressiveness of the pathogen in a range of densities that occurred naturally. The infectivity-related variable measured was the proportion of smutted plants at the end of each growing season; proportions were analysed using a generalised linear model with a binomial distribution considering the year, the density and their interaction as effects. The aggressiveness-related variables chosen were the number of smutted inflorescences per plant and per area, obtained over the last 2 years; they were analysed by means of ancova considering disease status (seeded or smutted), year, density and all the interactions between them. Although the disease is monocyclic, results showed clearly that infectivity increased with plant density. The number of inflorescences per plant was 1.5 times higher in smutted plants than in healthy plants throughout the range of densities. This variable declined when density increased, but as the infectivity increased at a higher rate, the aggressiveness also increased with density. The surprising results on infectivity are discussed in the context of current knowledge of plant-pathogen interaction dynamics, as well as neighbour effects on pathogen aggressiveness. Moreover, the results could be useful to develop weed biological control strategies.


Challenges and opportunities for cancer vaccines in the current NSCLC clinical scenario.
This review is aimed to focus on NSCLC as an emerging and promising model for active immunotherapy and the challenges for its inclusion in the current clinical scenario. Cancer vaccines for NSCLC have been focused as a therapeutic option based on the identification of a tumor hallmark and the active immunization with the related molecules that triggers cellular and/or humoral responses that consequently destroy or delay the rate of malignant progression. This therapeutic intervention in an established disease state has been aimed to impact into prolonging patient´s survival with ethically accepted quality of life. Understanding of relationship between structure and function in cancer vaccines is essential to interpret their opportunities to impact into prolonging survival and increasing quality of life in cancer patients. It is widely accepted that the failure of the cancer vaccines in the NSCLC scenario is related with its introduction in the advanced disease stages and poor performance status of the patients due to the combination of the tumor induced immunosuppression with the immune senescence. Despite first, second and emerging third line of onco-specific treatments the life expectancy for NSCLC patients diagnosed at advanced stages is surrounding the 12 months of median survival and in facts the today real circumstances are extremely demanding for the success inclusion of cancer vaccines as therapeutic choice in the clinical scenario. The kinetics of the active immunizations encompasses a sequential cascade of clinical endpoints: starting by the activation of the immune system, followed by the antitumor response and finalizing with the consequential impact on patients' overall survival. Today this cascade of clinical endpoints is the backbone for active immunization assessment and moreover the concept of cancer vaccines, applied in the NSCLC setting, is just evolving as a complex therapeutic strategy, in which the opportunities for cancer vaccines start from the selection of the target cancer hallmark, followed by the vaccine formulation and its platforms for immune potentiating, also cover the successful insertion in the standard of care, the chronic administration beyond progression disease, the personalization based on predictors of response and the potential combination with other targeted therapies.


Hepatic Artery Pseudoaneurysm Following Orthotopic Liver Transplantation: Increasing Clinical Suspicion for a Rare but Lethal Pathology.
BACKGROUND Despite an incidence of between 1% and 2%, the mortality rate in ruptured hepatic artery pseudoaneurysm after orthotopic liver transplantation approaches 69%. Our aim is to report operative and outcomes data for 7 patients with pseudoaneurysm following transplant at 1 institution, with emphasis on suspected risk factors for aneurysm formation. From these risk factors, we performed a systematic review to assess their clinical saliency. MATERIAL AND METHODS Using PRISMA guidelines, we completed a PubMed and online database review to gather studies addressing risk factors for pseudoaneurysm following transplant. We cross-compared infection, Roux-en-Y hepaticojejunostomy, bile leak, and primary sclerosing cholangitis as independent risk factors in order to identify concomitance between each and pseudoaneurysm. RESULTS The incidence of pseudoaneurysm was 0.94%. Of pseudoaneurysm patients, 77.8% had documented infection. Of these, 36.5% had Roux-en-Y hepaticojejunostomy and 33.3% had a documented bile leak. Infection was present in 70% of patients with Roux-en-Y hepaticojejunostomy, 84% of patients with bile leak, and 93% of patients with primary sclerosing cholangitis. CONCLUSIONS Roux-en-Y hepaticojejunostomy, bile leak, and primary sclerosing cholangitis are important risk factors for pseudoaneurysm in the setting of infection. Occurring together, these risk factors should heighten clinical suspicion for their formation in the postoperative period.


Nondestructive fluorescence lifetime imaging and time-resolved fluorescence spectroscopy detect cartilage matrix depletion and correlate with mechanical properties.
Tissue engineers utilize a battery of expensive, time-consuming and destructive techniques to assess the composition and function of engineered tissues. A nondestructive solution to monitor tissue maturation would reduce costs and accelerate product development. As a first step toward this goal, two nondestructive, label-free optical techniques, namely multispectral fluorescent lifetime imaging (FLIm) and time-resolved fluorescence spectroscopy (TRFS), were investigated for their potential in evaluating the biochemical and mechanical properties of articular cartilage. Enzymatic treatments were utilized to selectively deplete cartilage of either collagen or proteoglycan, to produce a range of matrix compositions. Samples were assessed for their optical properties using a fiber-coupled optical system combining FLIm and TRFS, their biochemical and mechanical properties and by histological staining. Single and multivariable correlations were performed to evaluate relationships among these properties. FLIm- and TRFS-derived measurements are sensitive to changes in cartilage matrix and correlate with mechanical and biochemical assays. Mean fluorescence lifetime values extracted from FLIm images (375-410 nm spectral band) showed strong, specific correlations with collagen content (R2 = 0.79, p < 0.001) and tensile properties (R2 = 0.45, p = 0.02). TRFS lifetime measurements centered at 520 nm (with a 5 nm bandwidth) possessed strong, specific correlations with proteoglycan content (R2 = 0.59, p = 0.001) and compressive properties (R2 = 0.71, p < 0.001). Nondestructive optical assessment of articular cartilage, using a combination of FLIm- and TRFS-derived parameters, provided a quantitative method for determining tissue biochemical composition and mechanical function. These tools hold great potential for research, industrial and clinical settings.


Impairment of cerebral autoregulation predicts delayed cerebral ischemia after subarachnoid hemorrhage: a prospective observational study.
Delayed cerebral ischemia (DCI) is a recognized contributor to unfavorable outcome after subarachnoid hemorrhage (SAH). Recent data challenge the concept of vasospasm as the sole cause of ischemia and suggest a multifactorial process with dysfunctional cerebral autoregulation as a component. We tested the hypothesis that early autoregulatory failure, detected using near-infrared spectroscopy-based index, TOxa and transcranial Doppler-based index, Sxa, can predict DCI. In this prospective observational study we enrolled consecutive patients with aneurysmal SAH that occurred <5 days from admission. The primary end point was the occurrence of DCI within 21 days of ictus. The predictive value of autoregulatory disturbances detected in the first 5 days was assessed using univarate proportional hazards model and a multivariate model. Ninety-eight patients were included. Univariate analysis demonstrated increased odds of developing DCI when early autoregulation failure was detected (odds ratio [OR], 7.46; 95% confidence interval [CI], 3.03-18.40 and OR, 4.52; 95% CI, 1.84-11.07 for Sxa and TOxa, respectively) but not TCD-vasospasm (OR, 1.36; 95% CI, 0.56-3.33). In a multivariate model Sxa and TOxa remained independent predictors of DCI (OR, 12.66; 95% CI, 2.97-54.07 and OR, 5.34; 95% CI, 1.25-22.84 for Sxa and TOxa, respectively). Disturbed autoregulation in the first 5 days after SAH significantly increases the risk of DCI. Autoregulatory disturbances can be detected using near-infrared spectroscopy and transcranial Doppler technologies.


Strategies for salivary cortisol collection and analysis in research with children.
Salivary cortisol has emerged in pediatric research as an easy-to-collect, relatively inexpensive, biologic marker of stress. Cortisol is highly variable and is responsive to a wide range of factors that should be considered when incorporating this measure into research with children. Strategies for sample collection include: (1) standardizing the time for sample collection, including baseline samples; (2) using consistent collection materials and methods; (3) controlling for certain drinks, foods, medications, and diagnoses; and (4) establishing procedures and protocols. Other strategies for laboratory analyses include: (1) selecting the appropriate assay and laboratory; (2) identifying units of measure and norms; and (3) establishing quality controls. These strategies control extraneous variables and produce reliable and valid salivary cortisol results.


Sialoblastoma: utility of Ki-67 and p53 as a prognostic tool and review of literature.
Sialoblastoma is a rare tumor of the salivary gland that commonly occurs in the parotid gland and occasionally in the sub-mandibular gland. The malignant potential of sialoblastoma has been documented in only 3 of 32 cases of sialoblastoma reported thus far. In the last 15 years, we have encountered 2 cases of sialoblastoma, in a newborn and in a 15-year-old boy, both arising within the parotid gland. Case 1 has been previously reported and although there were 2 recurrences, at 1 and 9 years post resection, it has shown benign biological behavior. Case 2 is unusual since the patient presented with metastases. We reviewed the 2 cases, including the 2 recurrences from the first case, for histologic and immunohistochemical differences. Although both cases showed similar cytomorphologic features, there was a significant difference in Ki-67 expression: 20% in case 1 (original tumor), <2% in case 1 (recurrent tumor), and nearly 70-80% in the recent malignant case. The difference is remarkable when combined with p53 expression, which was focally positive in the first case but diffusely positive in the second. This report highlights the potential utility of proliferation markers such as Ki-67 in concert with p53 expression to better predict the biological behavior of a rare but locally aggressive neoplasm.


Locomotor- and Reward-Enhancing Effects of Cocaine Are Differentially Regulated by Chemogenetic Stimulation of Gi-Signaling in Dopaminergic Neurons.
Dopamine plays a key role in the cellular and behavioral responses to drugs of abuse, but the implication of metabotropic regulatory input to dopaminergic neurons on acute drug effects and subsequent drug-related behavior remains unclear. Here, we used chemogenetics [Designer Receptors Exclusively Activated by Designer Drugs (DREADDs)] to modulate dopamine signaling and activity before cocaine administration in mice. We show that chemogenetic inhibition of dopaminergic ventral tegmental area (VTA) neurons differentially affects locomotor and reward-related behavioral responses to cocaine. Stimulation of Gi-coupled DREADD (hM4Di) expressed in dopaminergic VTA neurons persistently reduced the locomotor response to repeated cocaine injections. An attenuated locomotor response was seen even when a dual-viral vector approach was used to restrict hM4Di expression to dopaminergic VTA neurons projecting to the nucleus accumbens. Surprisingly, despite the attenuated locomotor response, hM4Di-mediated inhibition of dopaminergic VTA neurons did not prevent cocaine sensitization, and the inhibitory effect of hM4Di-mediated inhibition was eliminated after withdrawal. In the conditioned place-preference paradigm, hM4Di-mediated inhibition did not affect cocaine-induced place preference; however, the extinction period was extended. Also, hM4Di-mediated inhibition had no effect on preference for a sugar-based reward over water but impaired motivation to work for the same reward in a touchscreen-based motivational assay. In addition, to support that VTA dopaminergic neurons operate as regulators of reward motivation toward both sugar and cocaine, our data suggest that repeated cocaine exposure leads to adaptations in the VTA that surmount the ability of Gi-signaling to suppress and regulate VTA dopaminergic neuronal activity.


Similarity between brain activity at encoding and retrieval predicts successful realization of delayed intentions.
Remembering delayed intentions can be highly demanding. Accuracy in laboratory paradigms assessing prospective memory (PM) is typically well below ceiling, and failure to remember intended behaviors after a delay is a common occurrence in everyday life. However, relatively little is known of the potential differences in brain activity that distinguish successful versus unsuccessful PM. In this fMRI study, participants repeatedly encoded, stored, and then had the opportunity to retrieve intended behaviors while engaged in a distracting ongoing task. This yielded a success rate of approximately two thirds. Overall levels of brain activity distinguished successful versus unsuccessful trials at all three stages (encoding, storage, and retrieval), suggesting multiple neural determinants of PM success. In addition, the voxelwise similarity between patterns of brain activity at encoding and retrieval was greater for successful than unsuccessful trials. This was true even in posterior cingulate, which showed opposite patterns of signal change between encoding and retrieval. Thus, successful realization of delayed intentions may be associated with reinstatement of encoding context at the time of retrieval.


Colloidal Lignin Particles as Adhesives for Soft Materials.
Lignin has interesting functionalities to be exploited in adhesives for medicine, foods and textiles. Nanoparticles (NPs) < 100 nm coated with poly (L-lysine), PL and poly(L-glutamic acid) PGA were prepared from the laccase treated lignin to coat nanocellulose fibrils (CNF) with heat. NPs ca. 300 nm were prepared, β-casein coated and cross-linked with transglutaminase (Tgase) to agglutinate chamois. Size exclusion chromatography (SEC) and Fourier-transform infrared (FTIR) spectroscopy were used to characterize polymerized lignin, while zeta potential and dynamic light scattering (DLS) to ensure coating of colloidal lignin particles (CLPs). Protein adsorption on lignin was studied by quartz crystal microbalance (QCM). Atomic force microscopy (AFM) was exploited to examine interactions between different polymers and to image NPs with transmission electron microscopy (TEM). Tensile testing showed, when using CLPs for the adhesion, the stress improved ca. 10 and strain ca. 6 times compared to unmodified Kraft. For the β-casein NPs, the values were 20 and 8, respectively, and for the β-casein coated CLPs between these two cases. When NPs were dispersed in adhesive formulation, the increased Young's moduli confirmed significant improvement in the stiffness of the joints over the adhesive alone. Exploitation of lignin in nanoparticulate morphology is a potential method to prepare bionanomaterials for advanced applications.


Reducing population salt intake worldwide: from evidence to implementation.
Raised blood pressure is a major cause of cardiovascular disease, responsible for 62% of stroke and 49% of coronary heart disease. There is overwhelming evidence that dietary salt is the major cause of raised blood pressure and that a reduction in salt intake lowers blood pressure, thereby, reducing blood pressure-related diseases. Several lines of evidence including ecological, population, and prospective cohort studies, as well as outcome trials, demonstrate that a reduction in salt intake is related to a lower risk of cardiovascular disease. Increasing evidence also suggests that a high salt intake may directly increase the risk of stroke, left ventricular hypertrophy, and renal disease; is associated with obesity through soft drink consumption; is related to renal stones and osteoporosis; is linked to the severity of asthma; and is probably a major cause of stomach cancer. In most developed countries, a reduction in salt intake can be achieved by a gradual and sustained reduction in the amount of salt added to foods by the food industry. In other countries where most of the salt consumed comes from salt added during cooking or from sauces, a public health campaign is needed to encourage consumers to use less salt. Several countries have already reduced salt intake. The challenge now is to spread this out to all other countries. A modest reduction in population salt intake worldwide will result in a major improvement in public health.


Investigation of antileshmanial activities of Cuminum cyminum based green silver nanoparticles on L. tropica promastigotes and amastigotes in vitro.
Leishmaniasis is one of the most important parasitic diseases, which is caused by Leishmania species. Nowadays; although pentavalent antimonials are used as the basic treatment option for Cutaneous Leishmaniasis, high cost, toxicity and resistance of the parasites to the medication over time are some important limitations causing several problems in the treatment. In recent years, the progress in the field of green nanotechnology provides the development of green nanoparticle-based treatment methods for Cutaneous Leishmaniasis. The importance of green nanoparticles has gradually increased due to their special reductive, stabilizing, antioxidant and non-toxic properties. Although there are many studies based on green nanoparticles against Leishmania parasites, we have not found any research about antileishmanial activities of biosynthesized silver nanoparticles (Bio-AgNPs) using Cuminum cyminum L (Cumin) seed extract. Therefore for the first time in this study in vitro antileishmanial effects of Bio-AgNPs prepared from Cumin seed extract were examined on L. tropica promastigote and amastigote forms and their efficacies were compared with chemically synthetized AgNPs. During the experiments, antileishmanial effects of synthetized nanoparticles were determined on both promastigote and amastigote forms of Leishmania parasites by detecting different parameters such as proliferation, infection index and produced nitric oxide (NO) amounts from macrophages. According to the results, it was shown that Bio-AgNPs and AgNPs excessively inhibited L. tropica promastigotes and amastigotes by significantly decreasing proliferation rates of promastigotes and metabolic activities of amastigotes, as well. Moreover, infection index rates of macrophages revealed remarkable anti-amastigote performances of Bio-AgNPs. Besides, Bio-AgNPs stimulated macrophages to release NO to kill Leishmania parasites. Consequently, for the first time, Bio-AgNPs were found to be more effective on both forms of Leishmania parasites than AgNPs. Obtained results indicated high antileishmanial potential of green nanoparticles on L. tropica parasites, causative agents of Cutaneous Leishmaniasis. Thus, obtained results demonstrated that green nanoparticles can lead to the development of new, safer, stable and more effective antileishmanial formulations against Cutaneous Leishmaniasis.


Azimuthal multiple-beam interference effects with combinations of vortex beams.
We create a series of vortex beams consisting of positive and negative topological charges and develop a geometry where they interfere creating azimuthal multiple-beam interference effects. Usually the combination of two beams with opposite charges creates an azimuthal two-beam interference effect where the intensity varies sinusoidally with azimuthal angle. We combine several of these beams to create patterns where the interference becomes more sharply defined in the azimuthal direction and where destructive interference eliminates some of the intensity peaks. The process is complicated by the fact that the radii of the different vortex beams depend on the topological charge and the focal length of the Fourier lens. We generate a series of patterns where the different charges are focused with different focal lengths such that their radii agree. However to encode these, we encode patterns from the lower charges that have been Fresnel diffracted to the plane of the highest charge. These complicated patterns are encoded onto a liquid crystal display (LCD). Experimental results agree well with theory.


Endoscopic transnasal transclival odontoidectomy: a new approach to decompression: technical case report.
Endoscopic transnasal transclival resection of the odontoid process is less invasive than the standard transoral odontoidectomy. In this article, we describe our techniques, which are less invasive but provide successful decompression. From September 2004 to April 2007, three consecutive patients with basilar invagination and instability in the craniovertebral junction were enrolled. The causes for the invagination and instability included rheumatoid arthritis in two patients and trauma in one patient, and all patients presented with myelopathy and quadriparesis before intervention. All three patients underwent an endoscopic transnasal transclival approach for anterior decompression and resection of the displaced odontoid process and pannus to decompress the underlying medulla. Subsequently, they received occipitocervical fixation by lateral mass screws and bone fusion to ensure stability. Remarkable neurological recovery was observed after surgery in all patients, and no adverse effects were noted. Compared with the standard transoral approach, the transnasal transclival endoscopic approach for decompressing basilar invagination is a feasible and effective alternative that avoids common disadvantages like prolonged intubation, excessive tongue retraction, and the need for palatal incision.


[Psychopathology in families: an integral approach via the family outpatient clinic].
Psychiatric disorders run in families. To bridge the gap between child and youth psychiatry and adult psychiatry, GGZ inGeest has started screening parents of new registered children for psychopathology - and if indicated - offers parents treatment in the same department as their children. To examine the feasibility and usefulness of this procedure, to investigate how many parents agree to screening, further diagnostics and treatment, and to find out how many parents have in fact suffered from recent psychiatric problems. Prior to the children's first appointment, the parents were asked to complete a questionnaire, the Adult Self Report (ASR), about their own problems. If these scores were (sub)clinical, parents were invited to participate in a telephonic interview. This consisted of the Composite International Diagnostic Interview (CIDI) and Conners' Adult ADHD Rating Scales (CAARS). If the results indicate psychopathology, further psychiatric assessment and, if necessary, treatment is offered. The first response was 55.7% and, if indicated, most of the parents agreed on further diagnostics. On the ASR 2 out of 5 mothers (42.1%) and 1 out of 5 fathers (21.8%) reported problems that could point to a psychiatric disorder. According to the ASR, within this high-risk group 37% of the mothers met the criteria for an axis I diagnosis (less than one month earlier) compared to 70.6% of the fathers. A mood disorder was the primary diagnosis for women, whereas men most often suffered from an anxiety disorder. In total, 19.1% of the parents screened were suffering from recent psychopathology and 75% of this group agreed to receive mental health care (treatment at the family outpatient clinic or referred to another clinic). Implementation of the family outpatient clinic scheme is feasible. However, further efforts are needed in order to reach a larger group of parents, particularly fathers. The family outpatient clinic is useful because parents who suffer from psychopathology do not always receive mental health care. However, a randomised control trial is needed to determine whether parallel treatment of parents and children can improve the treatment outcome for children.


Resveratrol ameliorates renal damage, increases expression of heme oxygenase-1, and has anti-complement, anti-oxidative, and anti-apoptotic effects in a murine model of membranous nephropathy.
Idiopathic membranous nephropathy (MN) is an autoimmune-mediated glomerulonephritis and a common cause of nephrotic syndrome in adults. There are limited available treatments for MN. We assessed the efficacy of resveratrol (RSV) therapy for treatment of MN in a murine model of this disease. Murine MN was experimentally induced by daily subcutaneous administration of cationic bovine serum albumin, with phosphate-buffered saline used in control mice. MN mice were untreated or given RSV. Disease severity and pathogenesis was assessed by determination of metabolic and histopathology profiles, lymphocyte subsets, immunoglobulin production, oxidative stress, apoptosis, and production of heme oxygenase-1 (HO1). MN mice given RSV had significantly reduced proteinuria and a marked amelioration of glomerular lesions. RSV also significantly attenuated immunofluorescent staining of C3, although there were no changes of serum immunoglobulin levels or immunocomplex deposition in the kidneys. RSV treatment of MN mice also reduced the production of reactive oxygen species (ROS), reduced cell apoptosis, and upregulated heme oxygenase 1 (HO1). Inhibition of HO1 with tin protoporphyrin IX partially reversed the renoprotective effects of RSV. The HO1 induced by RSV maybe via Nrf2 signaling. Our results show that RSV increased the expression of HO1 and ameliorated the effects of membranous nephropathy in a mouse model due to its anti-complement, anti-oxidative, and anti-apoptotic effects. RSV appears to have potential as a treatment for MN.


Steroids as central regulators of organismal development and lifespan.
Larvae of the nematode Caenorhabditis elegans must choose between reproductive development and dauer diapause. This decision is based on sensing of environmental inputs and dauer pheromone, a small molecule signal that serves to monitor population density. These signals are integrated via conserved neuroendocrine pathways that converge on steroidal ligands of the nuclear receptor DAF-12, a homolog of the mammalian vitamin D receptor and liver X receptor. DAF-12 acts as the main switch between gene expression programs that drive either reproductive development or dauer entry. Extensive studies in the past two decades demonstrated that biosynthesis of two bile acid-like DAF-12 ligands, named dafachronic acids (DA), controls developmental fate. In this issue of PLoS Biology, Wollam et al. showed that a conserved steroid-modifying enzyme, DHS-16, introduces a key feature in the structures of the DAF-12 ligands, closing a major gap in the DA biosynthesis pathway. The emerging picture of DA biosynthesis in C. elegans enables us to address a key question in the field: how are complex environmental signals integrated to enforce binary, organism-wide decisions on developmental fate? Schaedel et al. demonstrated that pheromone and DA serve as competing signals, and that a positive feedback loop based on regulation of DA biosynthesis ensures organism-wide commitment to reproductive development. Considering that many components of DA signaling are highly conserved, ongoing studies in C. elegans may reveal new aspects of bile acid function and lifespan regulation in mammals.


Survival with an asymmetrical brain: advantages and disadvantages of cerebral lateralization.
Recent evidence in natural and semi-natural settings has revealed a variety of left-right perceptual asymmetries among vertebrates. These include preferential use of the left or right visual hemifield during activities such as searching for food, agonistic responses, or escape from predators in animals as different as fish, amphibians, reptiles, birds, and mammals. There are obvious disadvantages in showing such directional asymmetries because relevant stimuli may be located to the animal's left or right at random; there is no a priori association between the meaning of a stimulus (e.g., its being a predator or a food item) and its being located to the animal's left or right. Moreover, other organisms (e.g., predators) could exploit the predictability of behavior that arises from population-level lateral biases. It might be argued that lateralization of function enhances cognitive capacity and efficiency of the brain, thus counteracting the ecological disadvantages of lateral biases in behavior. However, such an increase in brain efficiency could be obtained by each individual being lateralized without any need to align the direction of the asymmetry in the majority of the individuals of the population. Here we argue that the alignment of the direction of behavioral asymmetries at the population level arises as an "evolutionarily stable strategy" under "social" pressures occurring when individually asymmetrical organisms must coordinate their behavior with the behavior of other asymmetrical organisms of the same or different species.


Overall efficacy of HPV-16/18 AS04-adjuvanted vaccine against grade 3 or greater cervical intraepithelial neoplasia: 4-year end-of-study analysis of the randomised, double-blind PATRICIA trial.
Cervical intraepithelial neoplasia grade 2 or greater (CIN2+) is the surrogate endpoint used in licensure trials of human papillomavirus (HPV) vaccines. Vaccine efficacy against CIN3+, the immediate precursor to invasive cervical cancer, is more difficult to measure because of its lower incidence, but provides the most stringent evidence of potential cancer prevention. We report vaccine efficacy against CIN3+ and adenocarcinoma in situ (AIS) in the end-of-study analysis of PATRICIA (PApilloma TRIal against Cancer In young Adults). Healthy women aged 15-25 years with no more than six lifetime sexual partners were included in PATRICIA, irrespective of their baseline HPV DNA status, HPV-16 or HPV-18 serostatus, or cytology. Women were randomly assigned (1:1) to receive an HPV-16/18 AS04-adjuvanted vaccine or a control hepatitis A vaccine via an internet-based central randomisation system using a minimisation algorithm to account for age ranges and study sites. The patients and study investigators were masked to allocated vaccine. The primary endpoint of PATRICIA has been reported previously. In the present end-of-study analysis, we focus on CIN3+ and AIS in the populations of most clinical interest, the total vaccinated cohort (TVC) and the TVC-naive. The TVC comprised all women who received at least one vaccine dose, approximating catch-up populations and including sexually active women (vaccine n=9319; control=9325). The TVC-naive comprised women with no evidence of oncogenic HPV infection at baseline, approximating early adolescent HPV exposure (vaccine n=5824; control=5820). This study is registered with ClinicalTrials.gov, number NCT00122681. Vaccine efficacy against CIN3+ associated with HPV-16/18 was 100% (95% CI 85·5-100) in the TVC-naive and 45·7% (22·9-62·2) in the TVC. Vaccine efficacy against all CIN3+ (irrespective of HPV type in the lesion and including lesions with no HPV DNA detected) was 93·2% (78·9-98·7) in the TVC-naive and 45·6% (28·8-58·7) in the TVC. In the TVC-naive, vaccine efficacy against all CIN3+ was higher than 90% in all age groups. In the TVC, vaccine efficacy against all CIN3+ and CIN3+ associated with HPV-16/18 was highest in the 15-17 year age group and progressively decreased in the 18-20 year and 21-25 year age groups. Vaccine efficacy against all AIS was 100% (31·0-100) and 76·9% (16·0-95·8) in the TVC-naive and TVC, respectively. Serious adverse events occurred in 835 (9·0%) and 829 (8·9%) women in the vaccine and control groups, respectively; only ten events (0·1%) and five events (0·1%), respectively, were considered to be related to vaccination. PATRICIA end-of-study results show excellent vaccine efficacy against CIN3+ and AIS irrespective of HPV DNA in the lesion. Population-based vaccination that incorporates the HPV-16/18 vaccine and high coverage of early adolescents might have the potential to substantially reduce the incidence of cervical cancer. GlaxoSmithKline Biologicals.


Residency training in neonatal circumcision: a pilot study and needs assessment.
Routine neonatal circumcision is one of the most commonly performed procedures in a neonate. Residents are expected to acquire the skills to properly evaluate the neonate and gain proficiency in performing circumcision despite significant variability in training. We performed a needs assessment to evaluate obstetric-gynecology residency training in neonatal circumcision. We performed an online self-assessment survey of obstetric-gynecology residents at Prentice Hospital, Chicago, from November 2008 to February 2009. Using images of uncircumcised penises residents were asked to identify which patients were candidates for routine neonatal circumcision. Of 36 obstetric-gynecology residents 27 responded to the survey. Most respondents planned to perform neonatal circumcision when in practice, 44% had no formal training in circumcision and most were comfortable performing routine neonatal circumcision. Overall respondents were less comfortable evaluating whether the a newborn penis could undergo circumcision safely. When presented with 10 pictures of penises and asked to determine whether the neonate should undergo circumcision, 0% of respondents correctly identified all contraindications to neonatal circumcision with an average of 42% of contraindications identified correctly. Of the respondents 77% listed practical experience as the first choice to learn a procedure with an online module preferred by 55% as the second choice. Although most residents feel competent to technically perform the procedure, they are not confident in their ability to judge the appropriate contraindications to neonatal circumcision. This needs assessment highlights the necessity for further curriculum development and formalized training in this domain.


The adipose tissue as a source of vasoactive factors.
Obesity is associated with increased incidence of cardiovascular mortality. However, the mechanisms that link increased fat mass with hypercholesterolemia, hypertension, endothelial dysfunction and coronary heart disease have not been fully elucidated. Unravelling the diverse neuroendocrine systems, which regulate energy balance and body fat has been a long-standing challenge in biology, with obesity as an increasingly important public health focus. Until recently, the adipocyte has been considered only a passive tissue for the storage of excess energy in the form of fat. However, there is now compelling evidence that adipocytes act as endocrine, secretory cells. It has been shown that several hormones, growth factors and cytokines are actually expressed in white adipose tissue. In a dynamic view of the adipocyte a wide range of signals emanates from white adipose tissue such as tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), and their respective soluble receptors. White adipose tissue also secretes important regulators of lipoprotein metabolism like lipoprotein lipase (LPL), apolipoprotein E (apoE) and cholesteryl ester transfer protein (CETP). The increasing number of products secreted by adipocytes also includes leptin, estrogen, angiotensinogen, plasminogen activator inhibitor-1 (PAI-1), tissue factor and transforming growth factor-beta (TGF-beta). Nitric oxide synthase (NOS) has been also reported to be expressed in white adipose tissue. Acylation stimulating protein (ASP), adipophilin, adipoQ, adipsin, monobutyrin, agouti protein and factors related to pro-inflammatory and immune processes have also been shown to be released by white adipocytes. Since blood vessels express receptors for most of the adipocyte-derived factors, adipose tissue seems to play a key role in cardiovascular physiology through the existence of a network of local and systemic signals. The current knowledge in this field will be reviewed in the broader perspective of cardiovascular physiology and pathophysiology.


A stochastic model for speciation by mating preferences.
Mechanisms leading to speciation are a major focus in evolutionary biology. In this paper, we present and study a stochastic model of population where individuals, with type a or A, are equivalent from ecological, demographical and spatial points of view, and differ only by their mating preference: two individuals with the same genotype have a higher probability to mate and produce a viable offspring. The population is subdivided in several patches and individuals may migrate between them. We show that mating preferences by themselves, even if they are very small, are enough to entail reproductive isolation between patches, and we provide the time needed for this isolation to occur as a function of the carrying capacity. Our results rely on a fine study of the stochastic process and of its deterministic limit in large population, which is given by a system of coupled nonlinear differential equations. Besides, we propose several generalisations of our model, and prove that our findings are robust for those generalisations.


Optical fuzzy systems based on efficient implementation of a fuzzy associative memory.
We propose efficient implementation of a fuzzy associative memory system based on a fuzzy-sets decomposition technique. The technique involves decomposing the input fuzzy sets of a fuzzy associative memory into orthogonal subsets so multiple fuzzy rules can be superimposed without loss of information. This technique can significantly reduce the optical hardware and computation needs of fuzzy associative memory systems.


The future of tobacco regulation: a response to a proposal for fundamental institutional change.
There is much debate about the future of tobacco regulation. Some have argued that to achieve maximum possible reduction in the harm tobacco causes, new regulatory models that involve fundamental institutional change must be adopted. The tobacco industry will continue to undermine tobacco control, and, so it is argued, we must change the way in which it operates if we are to be able to make the necessary progress. Such proposals may have an immediate appeal, especially for those frustrated that progress in tobacco control is not as rapid as they would hope--with all the avoidable loss of life and livelihoods that that entails--but they need to be carefully analysed and assessed, and much thought must be applied before spending time and resources advocating for them. One such proposal, advanced by Borland, involves the establishment of a monopsonistic statutory purchasing agency and wholesale distributor of tobacco products. Borland argues the relative benefits of such a model as compared to a more "conventional" model. On close examination, the benefits of the Borland proposal, as compared to the conventional model, appear overstated, and the Borland proposal introduces significant regulatory problems. There is still much that can be achieved in tobacco regulation through more conventional approaches, and within the "existing system". We should be wary of allowing the understandable allure of new or radical approaches to distract us from this reality.


Field Abundance Patterns and Odor-Mediated Host Choice by Clover Seed Weevils, Apion fulvipes and Apion trifolii (Coleoptera: Apionidae).
The clover seed weevils Apion fulvipes Geoffroy, 1785 and Apion trifolii L., 1768 (Coleoptera: Apionidae) cause major losses to seed production of white clover (Trifolium repens L.) and red clover (Trifolium pratense L.), respectively. Clover is important as animal forage and an alternative to inorganic fertilizers. Because clover is mainly pollinated by bees, the use of insecticides in management of these weevils is discouraged. To gain basic knowledge for development of alternative management strategies, we investigated weevil field abundance over two growing seasons, as well as feeding and olfactory host preferences by A. fulvipes and A. trifolii. Field trap catches in southern Sweden revealed that white clover was dominated by A. fulvipes and red clover by A. trifolii. For both weevil species, female catches were positively correlated to the number of clover buds and flowers in the field. In feeding and olfactory bioassays, females of A. fulvipes and A. trifolii showed a preference for T. repens and T. pratense, respectively. However, the feeding preference was lost when the antennae were removed, indicating a significant role of olfaction in host choice. Male weevils of both species did not show clear olfactory or feeding preferences for host plant species. The field study and laboratory bioassays demonstrate that, at least for female weevils, olfaction is important for selection of host plants. We discuss these novel results in the context of managing these important pests of clover by exploiting olfaction and behavioral attraction to host plant volatiles.


Motilimonas eburnea gen. nov., sp. nov., isolated from coastal sediment.
A novel Gram-stain-negative, non-spore-forming, facultatively anaerobic bacterium, designated YH6T, was isolated from marine sediment in Weihai, China. Cells of starin YH6T were motile, straight rods that formed ivory-white colonies on 2216E agar. Optimal growth occurred at 28-33 °C (range 15-37 °C), in the presence of 2-4 % (w/v) NaCl (range 1-8 %) and at pH 7.5-8.5 (range pH 6.5-9.0). The sole respiratory lipoquinone was Q-8, and the major fatty acids (>10 %) were C16 : 0 and summed feature 3 (C16 : 1ω7c/iso-C15 : 0 2-OH). The polar lipids profile of the novel strain consisted of phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol and several other unknown lipids (phospholipids, lipid and phosphoaminolipid). The G+C content of the genomic DNA was 46.5 mol%. The closest type strain phylogenetically to strain YH6T was Vibrio variabilis (92.99 % 16S rRNA gene sequence similarity) followed by Paramoritella alkaliphila (92.55 %), Pseudoalteromonas aurantia (92.20 %) and Pseudoalteromonas citrea (92.20 %). Phylogenetic analysis of the 16S rRNA gene sequence placed the novel strain in the order Alteromonadales, class Gammaproteobacteria. On the basis of the 16S rRNA gene sequence data as well as physiological and biochemical characteristics, we concluded that strain YH6T represents a novel species of a new genus. We propose the name of Motilimonas eburnea gen. nov., sp. nov. for this novel species. The type strain of the novel species is YH6T (=KCTC 42594T=MCCC 1H00122T).


Contractual health services performance agreements for responsive health systems: from conception to implementation in the case of Qatar.
Despite their use worldwide, strategy-based performance management is limited in the Eastern Mediterranean Region. This article explores Qatar's experience, the first from the Region, in implementing contractual agreements between healthcare providers and the regulator-Ministry of Public Health-to align strategy, performance and accountabilities. mixed-methods including tools development and pilot-testing, guided by performance management cycle with a focus on knowledge translation and key principles: feasibility; mandatory participation; participatory approach through Steering Committee. All public, private and semi-governmental hospitals and primary healthcare centers. (i) semi-structured interviews; (ii) review of 4982 indicators; (iii) Delphi technique for selecting indicators with > 80% agreement on importance and > 60% agreement on feasibility; (iv) capacity-building of providers and Ministry staff and 2-month pilot assessed by questionnaire with indicators scoring > 3 considered valid, reliable and feasible; and (v) 1-year grace period assessed by questionnaire. Approach strengths and challenges; Data collection and healthcare quality improvements. Contracts mandate reporting 25 hospital and 15 primary healthcare indicators to the regulator, which delivers confidential benchmarking reports to providers. Scorecards were discussed with the regulator for evidence-informed policymaking. The approach uncovered system-related challenges and learning for public and private sectors: providers commended the participatory approach (82%) and indicated that contracts enabled collecting valid and timely data (64%) and improved healthcare quality (55%). This experience provides insights for countries implementing performance management, responsive regulation and public-private partnerships. It suggests that contractual agreements can be useful, despite their mandatory nature, if clear principles are applied early on.


Synergistic blockade of mitotic exit by two chemical inhibitors of the APC/C.
Protein machines are multi-subunit protein complexes that orchestrate highly regulated biochemical tasks. An example is the anaphase-promoting complex/cyclosome (APC/C), a 13-subunit ubiquitin ligase that initiates the metaphase-anaphase transition and mitotic exit by targeting proteins such as securin and cyclin B1 for ubiquitin-dependent destruction by the proteasome. Because blocking mitotic exit is an effective approach for inducing tumour cell death, the APC/C represents a potential novel target for cancer therapy. APC/C activation in mitosis requires binding of Cdc20 (ref. 5), which forms a co-receptor with the APC/C to recognize substrates containing a destruction box (D-box). Here we demonstrate that we can synergistically inhibit APC/C-dependent proteolysis and mitotic exit by simultaneously disrupting two protein-protein interactions within the APC/C-Cdc20-substrate ternary complex. We identify a small molecule, called apcin (APC inhibitor), which binds to Cdc20 and competitively inhibits the ubiquitylation of D-box-containing substrates. Analysis of the crystal structure of the apcin-Cdc20 complex suggests that apcin occupies the D-box-binding pocket on the side face of the WD40-domain. The ability of apcin to block mitotic exit is synergistically amplified by co-addition of tosyl-l-arginine methyl ester, a small molecule that blocks the APC/C-Cdc20 interaction. This work suggests that simultaneous disruption of multiple, weak protein-protein interactions is an effective approach for inactivating a protein machine.


Activation of Cardiac Fibroblast Growth Factor Receptor 4 Causes Left Ventricular Hypertrophy.
Chronic kidney disease (CKD) is a worldwide public health threat that increases risk of death due to cardiovascular complications, including left ventricular hypertrophy (LVH). Novel therapeutic targets are needed to design treatments to alleviate the cardiovascular burden of CKD. Previously, we demonstrated that circulating concentrations of fibroblast growth factor (FGF) 23 rise progressively in CKD and induce LVH through an unknown FGF receptor (FGFR)-dependent mechanism. Here, we report that FGF23 exclusively activates FGFR4 on cardiac myocytes to stimulate phospholipase Cγ/calcineurin/nuclear factor of activated T cell signaling. A specific FGFR4-blocking antibody inhibits FGF23-induced hypertrophy of isolated cardiac myocytes and attenuates LVH in rats with CKD. Mice lacking FGFR4 do not develop LVH in response to elevated FGF23, whereas knockin mice carrying an FGFR4 gain-of-function mutation spontaneously develop LVH. Thus, FGF23 promotes LVH by activating FGFR4, thereby establishing FGFR4 as a pharmacological target for reducing cardiovascular risk in CKD.


Contact heterogeneity, rather than transmission efficiency, limits the emergence and spread of canine influenza virus.
Host-range shifts in influenza virus are a major risk factor for pandemics. A key question in the study of emerging zoonoses is how the evolution of transmission efficiency interacts with heterogeneity in contact patterns in the new host species, as this interplay influences disease dynamics and prospects for control. Here we use a synergistic mixture of models and data to tease apart the evolutionary and demographic processes controlling a host-range shift in equine H3N8-derived canine influenza virus (CIV). CIV has experienced 15 years of continuous transfer among dogs in the United States, but maintains a patchy distribution, characterized by sporadic short-lived outbreaks coupled with endemic hotspots in large animal shelters. We show that CIV has a high reproductive potential in these facilities (mean R(0) = 3.9) and that these hotspots act as refugia from the sparsely connected majority of the dog population. Intriguingly, CIV has evolved a transmission efficiency that closely matches the minimum required to persist in these refugia, leaving it poised on the extinction/invasion threshold of the host contact network. Corresponding phylogenetic analyses show strong geographic clustering in three US regions, and that the effective reproductive number of the virus (R(e)) in the general dog population is close to 1.0. Our results highlight the critical role of host contact structure in CIV dynamics, and show how host contact networks could shape the evolution of pathogen transmission efficiency. Importantly, efficient control measures could eradicate the virus, in turn minimizing the risk of future sustained transmission among companion dogs that could represent a potential new axis to the human-animal interface for influenza.


Toward prevention of Alzheimers disease--potential nutraceutical strategies for suppressing the production of amyloid beta peptides.
Alzheimers disease (AD) can be viewed as a vicious cycle in which excess production and deposition of amyloid beta (Abeta) peptides promote microglial activation, and the resultant production of inflammatory mediators further boosts Abeta production while inducing death and dysfunction of neurons. Abeta production is mediated by beta- and gamma-secretase activities; it is prevented by alpha-secretase activity, and insulin-degrading enzyme (IDE) catabolizes Abeta. High cellular cholesterol content increases Abeta synthesis by boosting beta-secretase activity; inhibition of cholesterol syntheses and/or stimulation of cholesterol export thus diminishes Abeta production. PPARgamma activity decreases Abeta production by promoting harmless catabolism of amyloid precursor protein while blocking the up-regulatory impact of cytokines on beta-secretase expression. Nitric oxide produced by the healthy cerebral microvasculature can suppress Abeta production by boosting expression of alpha-secretase while suppressing that of beta-secretase; conversely, cerebral ischemia provokes increased APP expression. Good insulin sensitivity and efficient brain insulin function protect by inhibiting gamma-secretase activity and increasing expression of IDE. The DHA provided by fish oil diminishes cerebral Abeta deposition in rodent AD models, for unclear reasons. Various measures which oppose microglial activation can inhibit up-regulation of beta-secretase and gamma-secretase by oxidants and cytokines, respectively. These considerations suggest that a number of nutraceutical or lifestyle measures may have potential for preventing or slowing AD: policosanol; 9-cis-beta-carotene; isomerized hops extract; DHA; measures which promote efficient endothelial NO generation, such as low-salt/potassium-rich diets, exercise training, high-dose folate, and flavanol-rich cocoa; chromium picolinate and cinnamon extract as aids for insulin sensitivity; and various agents which can oppose microglial activation, including vitamin D, genistein, and sesamin. The impact of these measures on Abeta production in rodent models of AD should be evaluated, with the intent of defining practical strategies for AD prevention.


An epidemic of yellow fever in central Brazil. 1972-1973. I. Epidemiological studies.
An epidemic of jungle yellow fever occurred in Goiás State, Brazil, between December 1972 and March 1973. Laboratory confirmed cases were observed in 36 counties located in the central and southern parts of the State. Seventy-one cases were proved, of which 44 were fatal. The diagnosis was made on the basis of pathology, serology, and virus isolation. Besides yellow fever, malaria and viral hepatitis were present, and in two fatal cases there was malarial pigment in the liver in addition to the specific lesions associated with yellow fever virus infection. The fact that male patients strikingly outnumbered females (9:1) and that young adults were predominantly affected indicates that transmission occurred mainly inside or adjacent to the forests. The lack of cases in urban areas can be attributed to the absence of Aedes aegypti in these areas. Yellow fever complement-fixing antibody in high titers was found in 18 of 1,201 (1.4%) persons living in eight counties of the affected area. This finding suggests that at least 21,000 persons out of the 1.5 million rural inhabitants of the three districts where the epidemic occurred had been infected by the virus. The epidemic subsided following an intensive vaccination campaign, and the last four cases were observed in March 1973.


Protective effects of garlic extract and vitamin C against in vivo cypermethrin-induced teratogenic effects in rat offspring.
Exposure of male (55.1 mg/kg b.wt. orally for 60 days) and/or pregnant female Wistar rats (55.1 mg/kg b.wt. orally at days 6-15 of gestation), to the insecticide cypermethrin (CYP); resulted in the development of a lot of external morphological deformities and visceral malformations in their offspring pubs, which signify the potential of such insecticide to induce reproductive toxicity and teratogenesis. Data cleared that CYP treatment induced significant increase in the percentages of post-implantation deaths, dwarf foeti and subcutaneous oedema beside significant decrease in percentages of live borne foeti and uterine implants. CYP also caused many visceral malformations among different treated groups including nasal, ophthalmic, cerebral, pulmonary, cardiac and renal malformations. Concomitant oral administration of garlic extract or vitamin C (5 days/week) to treated fathers and/or pregnant mothers with CYP provided significant reduction in the percentage of the foetal malformations induced by the insecticide, when compared with the control. The current study proves that garlic and ascorbic acid dampen the reproductive toxicity and/or teratogenicity of cypermethrin toxicity in rats; therefore might prove to be effective dietary supplements in developing countries where pesticide pollution is high.


C-Cl bond activation and catalytic hydrodechlorination of hexachlorobenzene by cobalt and nickel complexes with sodium formate as a reducing agent.
A benzyne cobalt complex, Co(η(2)-C6Cl4)(PMe3)3 (2), was generated from the reaction of hexachlorobenzene with 2 equiv. of Co(PMe3)4 through selective activation of two C-Cl bonds of hexachlorobenzene. Meanwhile, the byproduct CoCl2(PMe3)3 was also confirmed by IR spectra. The cobalt(II) complex, CoCl(C6Cl5)(PMe3)3 (1), as an intermediate in the formation of aryne complex 2, was also isolated by the reaction of hexachlorobenzene with the stoichiometric amount of Co(PMe3)4. Complex 2 could be obtained by the reaction of 1 with Co(PMe3)4. Under similar reaction conditions, the reaction of Ni(PMe3)4 with hexachlorobenzene afforded only a mono-(C-Cl) bond activation nickel(II) complex, NiCl(C6H5)(PMe3)2 (5). The expected benzyne nickel complex was not formed. The structures of complexes 2 and 5 were determined by X-ray single crystal diffraction. Successful selective hydrodechlorinations of hexachlorobenzene were studied and in the presence of Co(PMe3)4 or Ni(PMe3)4 as catalysts and sodium formate as a reducing agent pentachlorobenzene and 1,2,4,5-tetrachlorobenzene were obtained. The catalytic hydrodechlorination mechanism is proposed and discussed.


Effect of resveratrol on endothelial cell function: Molecular mechanisms.
The polyphenolic natural product resveratrol (RV), best known for its occurrence in grape skin and red wine, is considered a candidate drug for prevention and treatment of cardiovascular diseases. This review aims to summarize the molecular effects of RV on endothelial cells, which line the inner walls of blood vessels and play a key role in the development of those diseases. We describe how RV enhances endothelial nitric oxide production, improves endothelial redox balance and inhibits endothelial activation in response to pro-inflammatory and metabolic insults. Furthermore, we summarize effects of RV on endothelial senescence, apoptosis, endothelin-1 release, and endothelial progenitor cell function. As many of RV's actions seem to be mediated by SIRT₁, different mechanistic possibilities how RV may lead to SIRT₁ activation are discussed.


OH-initiated oxidation of sub-micron unsaturated fatty acid particles.
The heterogeneous reaction of OH radicals with sub-micron unsaturated fatty acid particles in the presence of H2O2 and O2 is studied to explore how surface OH addition reactions initiate chain reactions that rapidly transform the chemical composition of an organic particle. In the presence of 20.7 ppm [H2O2] in a 10% mixture of O2 in N2, the effective uptake coefficients of oleic acid, linoleic acid and linolenic acid are found to be 1.72 ± 0.08, 3.75 ± 0.18 and 5.73 ± 0.14, respectively. These effective uptake coefficients are larger than unity, providing clear evidence for particle-phase secondary chain chemistry. The effective uptake coefficient increases linearly with the number of C=C double bonds in the unsaturated fatty acid molecule. Elemental composition analysis reveals that there is an addition of, on average, 0.57 ± 0.02, 0.61 ± 0.01 and 0.73 ± 0.04 O atoms per reactive loss of oleic acid, linoleic acid and linolenic acid, respectively, which suggests that OH addition to the C=C double bond is not the sole reaction pathway that consumes the molecular species. These results suggest the potential presence of secondary reactions that consume the unsaturated fatty acid molecular species without increasing the particulate oxygen content. As the unsaturated fatty acid particles become more oxygenated, volatilization also becomes significant. The magnitudes of the effective uptake coefficients are found to be dependent on the concentrations of OH, O2 and H2O2 in the flow reactor. A plausible reaction mechanism is presented to show how surface OH addition reactions initiate chain reactions that rapidly transform an unsaturated organic particle's physicochemical properties.


An appraisal of indicators used to monitor the treated population in antiretroviral programmes in low-income countries.
Monitoring the progress of HIV programmes is vital, as services are scaled up to include increasing numbers in need of care. Globally, the presence of multiple donors at all levels of HIV care has produced vast monitoring systems. Within HIV-treatment programmes in low and middle-income countries, directly assessing long-term outcomes such as survival is problematic, so indicators are used to monitor the progress of the treated population. However, the internal, external, construct validity and predictive value of current indicators have never been evaluated. Although the burden on facility staff compiling routine monitoring reports is vast, there is uncertainty as to which indicators best monitor patient progress. This burden will grow as increasing numbers of life-cohorts are created for monitoring purposes leading to data inaccuracies and compromising the internal validity of reported indicators. Furthermore, a number of fundamental indicators, including survival and retention, may not capture the construct they intend to measure, compromising the ability of programme managers to obtain reliable estimates regarding the welfare of their population in care. It is not known which indicators can predict the longer-term outcome of the patient population, and as such, can enable managers to respond to predictors of failure early. An evaluation of current indicators is urgently needed to ensure that reported facility-level data accurately reflect the welfare of the treated population and comparisons of programme performance are meaningful.


GV1001 immunotherapy ameliorates joint inflammation in a murine model of rheumatoid arthritis by modifying collagen-specific T-cell responses and downregulating antigen-presenting cells.
This study investigated whether GV1001 may be useful for treating rheumatoid arthritis (RA). Two collagen-induced arthritis (CIA) experiments showed that therapeutic, but not preventive, GV1001 treatment reduced the severity of joint inflammation in CIA. The third CIA experiment indicated that, compared to vehicle treatment, therapeutic GV1001 treatment was associated with a significantly smaller area under the curve for the overall clinical joint score over the 98day observation period (p<0.05). GV1001 treatment was also associated with lower Day 98 serum IL-6 levels (p<0.01) and histological joint scores (p<0.05). Moreover, splenocytes harvested from the GV1001-treated mice exhibited lower basal and collagen-stimulated production of IFN-γ and IL-6 on Days 49 and 98 than the splenocytes from vehicle-treated mice. The fourth and fifth experiments indicated that earlier treatment resulted in a better response. In addition, human (THP-1) and murine (RAW 264.7) macrophages and fibroblast-like synoviocytes (FLS) from RA patients were used for in vitro analyses. GV1001 treatment of lipopolysaccharide-stimulated macrophages derived from THP-1 and RAW 264.7 monocytes significantly reduced TNF-α and IL-6 secretion (THP-1: all p<0.05; RAW 264.7: all p<0.01). However, GV1001 treatment did not affect IL-6 expression in TNFα-stimulated RA FLS. GV1001 reduced the clinical joint scores, serum IL-6 levels, and histological joint scores of mice with CIA. In addition, GV1001 lowered the collagen-stimulated IFN-γ and IL-6 production of murine T-cells and reduced the TNF-α and IL-6 production of macrophages in vitro. Thus, GV1001 may ameliorate joint inflammation by modifying T-cell reactions to the triggering autoantigen and by reducing macrophage cytokine production.


The role of the gut microbiota in energy metabolism and metabolic disease.
Obesity is now classically characterized by a cluster of several metabolic disorders, and by a low grade inflammation. The evidence that the gut microbiota composition can be different between healthy and or obese and type 2 diabetic patients has led to the study of this environmental factor as a key link between the pathophysiology of metabolic diseases and the gut microbiota. Several mechanisms are proposed linking events occurring in the colon and the regulation of energy metabolism, such as i.e. the energy harvest from the diet, the synthesis of gut peptides involved in energy homeostasis (GLP-1, PYY...), and the regulation of fat storage. Moreover, the development of obesity and metabolic disorders following a high-fat diet may be associated to the innate immune system. Indeed, high-fat diet feeding triggers the development of obesity, inflammation, insulin resistance, type 2 diabetes and atherosclerosis by mechanisms dependent of the LPS and/or the fatty acids activation of the CD14/TLR4 receptor complex. Importantly, fat feeding is also associated with the development of metabolic endotoxemia in human subjects and participates in the low-grade inflammation, a mechanism associated with the development of atherogenic markers. Finally, data obtained in experimental models and human subjects are in favour of the fact that changing the gut microbiota (with prebiotics and/or probiotics) may participate in the control of the development of metabolic diseases associated with obesity. Thus, it would be useful to find specific strategies for modifying gut microbiota to impact on the occurrence of metabolic diseases.


Migration of levonorgestrel IUS in a patient with complex medical problems: what should be done?
Patients with complex medical problems should be counselled about the need for highly effective contraception. As failure resulting in pregnancy, could cause significant morbidity and mortality. The LNG-IUS has gained great popularity and generally has a low side effect profile; however, perforation of the uterus and migration of the device is a potentially serious complication known to be associated with its use. The current accepted management is removal of the device from the abdominal cavity in order to prevent further morbidity. However this is not always a simple matter in patients who have complex medical problems and who are deemed unfit for surgery. Each time the patient comes for renewal of the contraceptive method, clinicians need to reassess the risks and benefits. This is particularly relevant in patients who have complex medical problems where special attention needs to be given, not only to immediate risks but also to long-term ones. Careful individualised counselling and consideration are paramount and perhaps it would have been prudent to discuss vasectomy with this patient and her husband (as the first line of contraception), as this may have avoided the ensuing complications arising from the chosen method.


The Analysis of Spirituality of Patients with HIV/AIDS in Taking Lessons and Self-Acceptance.
Nursing care on the spiritual aspect is focusing on the patients' acceptance of their diseases; thus, people living with HIV (PLWH) are able to accept their diseases and are able to take the lesson. PLWH do not only deal with the condition of the disease but also by discriminative social stigma. The aim of this study was to explore, describe, and interpret the experience of spirituality to self-acceptance in patients with HIV/AIDS. This research is a qualitative approach by descriptive phenomenology of participants involving as many as 10 people, consisting of 5 men and 5 women. All paticipants are muslim with the education level range from junior high school to university. The ages ranged from 29 to 46 years. This research identified the two themes which are: (1) being able to take the lesson from their diseases, and (2) self acceptance as people living with HIV-AIDS. There were 10 participants participated in this study. A method of in-depth interviews and observation is a help of data collection. Data analysis used was Creswell method. Results of the study suggested the patients to get motivated and to develop aspects of spirituality so that it can help to ease in the process of self acceptance, asgetting closer to God through pray, read the Quran, fasting, etc.


The reconstruction aides.
The reconstruction aides, civilian women who served in World War I, are credited with an influential role in the development of occupational therapy. Their task was to provide treatment in the form of occupation to enable servicemen suffering from wounds or battle neurosis to return to the battlefront. Although some occupational therapy aides were occupational therapists, many were teachers, artists, and craftspersons. This paper traces the history of the reconstruction aides, describes the women who served, and recounts their experiences. The relationships between reconstruction aides and other professions suggest the origins of current problems of professional identity and role delineation.


Cervical Pregnancy Masquerading as an Incomplete Abortion- A Learning Lesson.
Cervical pregnancy is a rare site of ectopic pregnancy compared to tubal. The trophoblast implant into the cervical tissue and become a potentially dangerous site of torrential haemorrhage. The widespread use of Ultrasonography (USG) has led to a dramatic increase in the detection rates of extra uterine pregnancy. We hereby report an interesting case of extra uterine pregnancy with a unique situation where only a high index of suspicion prevented an iatrogenic mishap. The patient was referred as a case of incomplete abortion with a documented report of minimal retained products. Because of a high index of suspicion a serum beta Human chorionic gonadotropin (hCG) was sent prior to deciding for discharge. This indeed turned as a major change in the diagnosis of the case as the serum beta hCG was elevated. On re-evaluation, we diagnosed it as a case of cervical pregnancy which was successfully managed medically.


Localising and tailoring research evidence helps public health decision making.
Published research evidence is typically not readily applicable to practice but needs to be actively mobilised. This paper explores the mechanisms used by information professionals with a specific knowledge mobilisation role to make evidence useful for local decision making and planning of public health interventions. Data are drawn from a NIHR project that studied how, when, where and by whom published research evidence is used in commissioning and planning across two sites (one in England and one in Scotland). Data included 11 in-depth interviews with information professionals, observations at meetings and documentary analysis. Published research evidence is made fit for local commissioning and planning purposes by information professionals through two mechanisms. They localise evidence (relate evidence to local context and needs) and tailor it (present actionable messages). Knowledge mobilisation roles of information professionals are not recognised and researched. Information professionals contribute to the 'inform' and 'relational' functions of knowledge mobilisation; however, they are less involved in improving the institutional environment for sustainable knowledge sharing. Information professionals are instrumental in shaping what evidence enters local decision making processes. Identifying and supporting knowledge mobilisation roles within health libraries should be the focus of future research and training.


Prostate cancer in men using testosterone supplementation.
Although an association between testosterone supplementation and the development of prostate cancer is unproven, a recent increase in the use of this therapy has reopened the debate about its safety in men at risk for prostate cancer. To increase awareness of this risk, we report on a series of patients in whom clinically significant prostate cancer developed and was presumed to be related to exogenous testosterone use. The medical records of 6 urology practices were reviewed to identify men undergoing testosterone supplementation for sexual dysfunction or "rejuvenation " who were found to have prostate cancer after initiation of exogenous testosterone supplementation. Cases were analyzed to determine clinical and pathological parameters characterizing the presentation of prostate cancer. A total of 20 men were diagnosed with prostate cancer after initiation of testosterone therapy. Prostate cancer was detected within 2 years of testosterone initiation in 11 men (55%) and from 28 months to 8 years in the remainder. The tumors were of moderate and high grade, being Gleason sum 6, 7 and 8 to 10 in 9 (45%), 6 (30%) and 5 (25%) men, respectively. Median serum prostate specific antigen (PSA) concentration at diagnosis tended to be low at 5.1 ng/ml (range 1.1 to 329.0) and digital rectal examination generally proved more sensitive than PSA assays in detecting the cancer. Patients seen by nonurologist physicians were monitored less often for prostate cancer during testosterone use than those followed by urologists. Prostate cancer may become clinically apparent within months to a few years after the initiation of testosterone treatment. Digital rectal examination is particularly important in the detection of these cancers. Physicians prescribing testosterone supplementation and patients receiving it should be cognizant of this risk, and serum PSA testing and digital rectal examination should be performed frequently during treatment.


Relationship Between Calcium Intake and Impaired Activities of Daily Living in a Japanese Population: NIPPON DATA90.
Major reasons for long-term care insurance certification in Japan are stroke, dementia, and fracture. These diseases are reported to be associated with calcium intake. This study examined the association between calcium intake and impaired activities of daily living (ADL) using the data from NIPPON DATA90, consisting of representative sample of the Japanese population. A population-based nested case-control study was performed. A baseline survey was conducted in 1990, followed by ADL surveys of individuals ≥65 years old in 2000. Individuals with impaired ADL and selected age- and sex-matched controls were then identified. We obtained 132 pairs. Calcium intake was energy-adjusted using the residual method. The association between calcium intake and impaired ADL was examined using conditional logistic regression models. To assess the accuracy of the estimates, we conducted bootstrap analyses. The adjusted odds ratios (ORs) for impaired ADL compared with the group with a calcium intake of <476 mg/day were 0.72 (95% confidence interval [CI], 0.37-1.40) for the 476-606 mg/day group and 0.44 (95% CI, 0.21-0.94) for the ≥607 mg/day group in 2000 (P for linear trend = 0.03). After the bootstrap analyses, the inverse relationship unchanged (median OR per 100-mg rise in calcium intake, 0.87 [1,000 resamplings]; 95% CI, 0.76-0.97). After bootstrap analyses, calcium intake was inversely associated with impaired ADL 10 years after the baseline survey.


The experiences of close relatives living with a person with serious chronic illness.
Living with serious chronic illness has a directly threatening effect on the life course of healthy relatives. The aim of this study was to elucidate the meaning of close relatives' experiences of living with a person with serious, chronic illness. Using a narrative approach, the authors interviewed 13 spouses and 1 daughter about the meaning of their experiences. They used a phenomenological hermeneutic method to interpret the interview text. The structural analysis revealed three major themes: A Shrinking Life, Forced to Take Responsibility, and Struggling to Keep Going. Close relatives in this study seemed to be living lives characterized by a reduced sense of individual freedom and an increased sense of responsibility for the care of the ill person. It means struggling to obtain the strength to manage their duty and an inner sense of community and of solidarity. The authors interpret the findings in the light of the works of philosophers who have described ethical demands and natural love between people.


AKAP3 selectively binds PDE4A isoforms in bovine spermatozoa.
Cyclic AMP plays an important role in regulating sperm motility and acrosome reaction through activation of cAMP-dependent protein kinase A (PKA). Phosphodiesterases (PDEs) modulate the levels of cyclic nucleotides by catalyzing their degradation. Although PDE inhibitors specific to PDE1 and PDE4 are known to alter sperm motility and capacitation in humans, little is known about the role or subcellular distribution of PDEs in spermatozoa. The localization of PKA is regulated by A-kinase anchoring proteins (AKAPs), which may also control the intracellular distribution of PDE. The present study was undertaken to investigate the role and localization of PDE4 during sperm capacitation. Addition of Rolipram or RS25344, PDE4-specific inhibitors significantly increased the progressive motility of bovine spermatozoa. Immunolocalization techniques detected both PDE4A and AKAP3 (formerly known as AKAP110) in the principal piece of bovine spermatozoa. The PDE4A5 isoform was detected primarily in the Triton X-100-soluble fraction of caudal epididymal spermatozoa. However, in ejaculated spermatozoa it was seen primarily in the SDS-soluble fraction, indicating a shift in PDE4A5 localization into insoluble organelles during sperm capacitation. AKAP3 was detected only in the SDS-soluble fraction of both caudal and ejaculated sperm. Immunoprecipitation experiments using COS cells cotransfected with AKAP3 and either Pde4a5 or Pde4d provide evidence that PDE4A5 but not PDE4D interacts with AKAP3. Pulldown assays using sperm cell lysates confirm this interaction in vitro. These data suggest that AKAP3 binds both PKA and PDE4A and functions as a scaffolding protein in spermatozoa to regulate local cAMP concentrations and modulate sperm functions.


Post-Operative Radiotherapy for Soft Tissue Sarcoma of the Anterior Compartment of the Thigh: Should the Sartorius Muscle be Included?
The clinical target volume (CTV) of post-operative radiotherapy for soft tissue sarcoma of the limbs conventionally includes the whole of the transverse cross-section of the affected anatomical compartment. In the anterior thigh sartorius appears to lie within its own fascial compartment and can be safely excluded. We investigated the potential impact of omitting sartorius from the anterior muscle compartment on patients with soft tissue sarcoma of the thigh. We used the planning CT data from six patients who had previously received post-operative radiotherapy for soft tissue sarcoma of the thigh. The anterior compartments were outlined twice, initially including and then excluding the sartorius muscle. The volumes of the anterior compartment (i.e., the CTVs), both with and without sartorius, and the corresponding planning target volumes (PTVs) were calculated. Treatment plans were prepared for each PTV. For both volumes the unirradiated normal tissue corridor was outlined on each CT slice. The volume and circumference of the unirradiated corridor were then calculated. For all six patients there was an important improvement in normal tissue sparing by excluding sartorius. The mean reduction in volume of the anterior compartment when sartorius was excluded was 10% (95% Confidence Interval 8-12%), whilst the mean decrease in PTV was 11% (95% CI 7-14%). There was a substantial increase in the volume of the unirradiated normal tissue corridor, with a mean value of 77% (95% CI 41-114%) when sartorius was excluded. In addition, the percentage increase in the size of the unirradiated normal tissue corridor, expressed as a percentage of the whole leg circumference, was 10% (95% CI 8-13%). When sartorius was included in the anterior compartment, the circumference of the unirradiated corridor was less than one-third of the whole leg circumference in four of the six patients. When sartorius was excluded, the circumference of the unirradiated corridor was greater than one-third of the leg circumference over the entire length of the target volume in all patients. It is essential to know the anatomy of the sartorius muscle to be able to exclude it from the anterior compartment. The increase in the size of the normal tissue corridor when sartorius is excluded should deliver clinical advantage by decreasing the normal tissue adverse effects.


Pharmacogenomic profiling of an oxidative stress-mediated spongiform encephalopathy.
The majority of cellular superoxide is generated in the mitochondria as a by-product of normal oxidative metabolism. In the mitochondria, superoxide is detoxified by manganese superoxide dismutase (SOD2). Mice lacking SOD2 demonstrate a multifaceted neonatal lethal phenotype, including a spongiform encephalopathy that is preventable through antioxidant treatment. The molecular events behind the observed pathology in the cortex of these mice are unknown. We hypothesized that the lack of SOD2 would result in significant changes in cortical gene expression and that therapeutically beneficial antioxidant treatment would normalize the expression of some genes, providing insight into the mechanism by which mitochondrial oxidative stress results in neurodegeneration. We report the identification of gene expression profiles associated with this paradigm, which characterize the degree of response to the pharmacologic intervention. We have identified specific pathways targeted by endogenous oxidative stress, including glutathione metabolism, iron metabolism, and cell-survival pathways centering on the kinase AKT. The normalization of expression of some of these pathways by antioxidant treatment suggests approaches to treating disease in which endogenous oxidative stress plays a role.


Provider determinants of emergency contraceptive counseling and provision in Kenya and Ethiopia.
In a majority of sub-Saharan African countries, counseling and provision of emergency contraception (EC) lag behind that of developed countries. As policymakers expand EC programs in the region, an understanding of provider knowledge and bias regarding EC is critical. Using data from recent surveys of Kenyan and Ethiopian health care providers in bivariate analyses and multivariate logit regression models, this study assesses whether variation in provider knowledge and bias regarding EC is associated with variation in EC counseling and provision. Survey results indicate that 54% and 31% of Kenyan and Ethiopian providers, respectively, display strong EC counseling behavior, while 61% and 55%, respectively, report having ever provided EC. Bivariate and multivariate results show that, in Kenya, increased EC counseling and provision behaviors are associated with higher levels of provider knowledge. Training on EC can increase provider knowledge and lower bias, leading to improved access for clients.