Phenotypic shift in Wolbachia virulence towards its native host across serial horizontal passages.
Vertical transmission mode is predicted to decrease the virulence of symbionts. However, Wolbachia, a widespread vertically transmitted endosymbiont, exhibits both negative and beneficial effects on arthropod fitness. This 'Jekyll and Hyde' behaviour, as well as its ability to live transiently outside host cells and to establish new infections via horizontal transmission, may reflect the capacity of Wolbachia to exhibit various phenotypes depending on the prevailing environmental constraints. To study the ability of Wolbachia to readily cope with new constraints, we forced this endosymbiont to spread only via horizontal transmission. To achieve this, we performed serial horizontal transfers of haemolymph from Wolbachia-infected to naive individuals of the isopod Armadillidium vulgare. Across passages, we observed phenotypic changes in the symbiotic relationship: (i) The Wolbachia titre increased in both haemolymph and nerve cord but remained stable in ovaries; (ii) Wolbachia infection was benign at the beginning of the experiment, but highly virulent, killing most hosts after only a few passages. Such a phenotypic shift after recurrent horizontal passages demonstrates that Wolbachia can rapidly change its virulence when facing new environmental constraints. We thoroughly discuss the potential mechanism(s) underlying this phenotypic change, which are likely to be crucial for the ongoing radiation of Wolbachia in arthropods.


Calcitonin gene-related peptide immunoreactivity in spinal spheroids in motor neuron disease.
The spinal cords from seven autopsy cases of sporadic motor neuron disease (MND) and two controls were immunohistochemically examined using antibodies directed to calcitonin gene-related peptide (CGRP) and to neurofilament proteins (Nf). CGRP immunoreactivity was observed in the posterior horns, especially in the laminae I and II, of all the spinal cords examined. In MND cases, in addition, a considerable number of spheroids in the anterior horns were labelled with the antibody. In some spheroids, their entire area was homogeneously immunostained, whereas in others the immunoreactivity was confined to a focal area(s) within the profile of the spheroids and between these two forms of staining several variations of the staining patterns were seen. The anti-Nf intensely and homogeneously labelled all spheroids and there was no appreciable difference in the Nf-immunoreactive pattern between CGRP-positive and -negative spheroids. It is possible that the accumulation of CGRP in spheroids may result from entrapment of CGRP during the anterograde axonal transport, causing loss of CGRP at the neuromuscular junction and producing weakness and atrophy of the muscles.


Auriculotherapy used to manage orthodontic pain: a randomized controlled pilot study.
Several methods are commonly used to decrease orthodontic pain, but versatile tools and standardized protocols are still lacking. In response to the need for alternatives to conventional analgesic methods, this study evaluates the analgesic effects of auriculotherapy (AT) during the first three months of fixed orthodontic treatment. A sample of 36 subjects was selected, with patients randomly allocated into two homogeneous groups, Study Group (SG) and Control Group (CG), depending on the application/non-application of AT. Patients rated their pain scores monthly from 0 to 10, on visual analogue scales (VAS) at the time of bonding (T0) and again at two appliance adjustments (T1 and T2). At each of these treatment phases, VAS was applied in six different time moments (TM): immediately before, immediately after, after 4 hours, after 8 hours, after 24 hours, and after 72h hours. Descriptive statistical analysis, a Student's t-test, and a Chi-square test were applied to the collected data (statistical significance for p< 0.05). SG patients reported lower pain levels than CG patients, both at T0, T1 and T2. Moreover, average pain intensity values were lower in the SG for all TM analyzed, with the t-test significant (p< 0.05) for most TMs. AT was effective in the pain treatment of patients with fixed orthodontic appliances. Further studies are needed with a sham control group to confirm the validity of these results.


Female adnexal tumor of probable wolffian origin: a clinicopathological case report and a possible new treatment.
Female adnexal tumors of probable wolffian origin (FATWOs) are rare tumors arising in the broad ligament from the remnants of the mesonephric duct. We report a case of recurrent disease. A 15-year-old girl who presented with a painful pelvic mass underwent a laparotomy with tumor resection. Pathology findings confirmed a FATWO. The tumor recurred within 2 years and was treated with multiple chemotherapy regimens, including a platinum-based drug, and surgery for progressive disease. The tumor was positive for c-kit oncogene (CD 117). Gleevac therapy, a tyrosine kinase inhibitor, was prescribed, and she developed severe persistent lower abdominal pain 2 months later. She underwent a hysterectomy and debulking of retroperitoneal masses. Pathology showed evidence of tumor necrosis, suggesting a possible beneficial effect, and she was recommenced on Gleevac in an effort to prevent recurrences. She is currently asymptomatic, without evidence of disease 10 months after surgery, continuing on Gleevac therapy. FATWOs are very rare tumors. Most cases are benign but have the potential to recur and metastasize. There is limited knowledge about the optimal treatment for this neoplasm. Our patient's favorable response to Gleevac therapy supports the concept of targeted molecular therapy in patients with c-kit-positive FATWO tumors.


Incidence and outcomes in acute kidney injury: a comprehensive population-based study.
Epidemiological studies of acute kidney injury (AKI) and acute-on-chronic renal failure (ACRF) are surprisingly sparse and confounded by differences in definition. Reported incidences vary, with few studies being population-based. Given this and our aging population, the incidence of AKI may be much higher than currently thought. We tested the hypothesis that the incidence is higher by including all patients with AKI (in a geographical population base of 523,390) regardless of whether they required renal replacement therapy irrespective of the hospital setting in which they were treated. We also tested the hypothesis that the Risk, Injury, Failure, Loss, and End-Stage Kidney (RIFLE) classification predicts outcomes. We identified all patients with serum creatinine concentrations > or =150 micromol/L (male) or > or =130 micromol/L (female) over a 6-mo period in 2003. Clinical outcomes were obtained from each patient's case records. The incidences of AKI and ACRF were 1811 and 336 per million population, respectively. Median age was 76 yr for AKI and 80.5 yr for ACRF. Sepsis was a precipitating factor in 47% of patients. The RIFLE classification was useful for predicting full recovery of renal function (P < 0.001), renal replacement therapy requirement (P < 0.001), length of hospital stay [excluding those who died during admission (P < 0.001)], and in-hospital mortality (P = 0.035). RIFLE did not predict mortality at 90 d or 6 mo. Thus the incidence of AKI is much higher than previously thought, with implications for service planning and providing information to colleagues about methods to prevent deterioration of renal function. The RIFLE classification is useful for identifying patients at greatest risk of adverse short-term outcomes.


Induction of G₂/M Arrest by Berberine via Activation of PI3K/Akt and p38 in Human Chondrosarcoma Cell Line.
Berberine is a clinically important natural isoquinoline alkaloid found in many medicinal herbs. Berberine has been shown to have many pharmacological effects including antimicrobial, antitumor, and anti-inflammatory activities. However, the effects and mechanism of action of berberine have not been studied in chondrosarcoma. Therefore, the effects of berberine on proliferation in a human chondrosarcoma cell line (HTB-94) were investigated. Berberine inhibited cell proliferation in a concentration-dependent manner. We also determined that inhibition of cell proliferation by berberine occurred via G2/M phase arrest in HTB-94 cells. Berberine induced cell cycle arrest at the G2/M phase by upregulation of p53 and p21 expression and suppressed cyclin B1, cyclin-dependent kinase 1 (cdc2), cdc25c, and phosphorylated retinoblastoma tumor-suppressor protein (pRb) expression. In addition, berberine stimulated phosphorylation of protein kinase B (Akt) and p38 kinase. Inhibition of phosphatidylinositol 3-kinase (PI3K)/Akt with LY294002 (LY) and p38 kinase with SB203580 (SB), respectively, decreased berberine-induced p53 and p21 expression and restored cell proliferation and expression of cyclin B1, cdc2, cdc25c, and pRb cell cycle progression proteins. These results suggest that berberine-induced inhibition of cell proliferation by cell cycle arrest at the G2/M phases was regulated through PI3K/Akt and p38 kinase pathways in HTB-94 chondrosarcoma cells.


Iatrogenic causes of an ICH: OAT therapy.
Current understanding of oral anticoagulant treatment and related intracerebral haemorrhage remains rather limited as compared to that of spontaneous intracerebral haemorrhage. Although intracerebral haemorrhage is the most serious complication of oral anticoagulant treatment, standardized treatment guidelines are still lacking. The currently employed treatments are aimed at normalization of the iatrogenic coagulation impairment, and are not based on randomized controlled trials evidence. Since most patients with oral anticoagulant treatment-intracerebral haemorrhage are at high risk of cardio-embolism and often myocardial infarction, it is uncertain whether the use of procoagulant treatments for oral anticoagulant treatment-intracerebral haemorrhage may increase their risk of thrombotic complications. Patients who receive chronic oral anticoagulant treatment urgently require effective treatments for acute oral anticoagulant treatment-intracerebral haemorrhage, and therefore controlled clinical trials are needed.


Object play in infants with autism: methodological issues in retrospective video analysis.
Using a taxonomy of object play, this study describes methodological issues in using retrospective video analysis and computer-based coding as a research tool for early identification of autism. Home videos of 32 infants with autism (n= 11), developmental delay (n= 10), and typical development (n= 11) were edited and analyzed for duration and highest level of object play in four hierarchical categories (exploratory, relational, functional, symbolic) using The Observer 3.0. The three groups had similar levels of engagement with objects, and no statistically significant differences in duration of exploratory play. Higher levels of play were rarely evident at 9-12 months, however, the highest level achieved (functional play) was apparent only in the typical group. This study provides the first naturalistic investigation of object play skills in infants with autism ages 9-12 months. It also demonstrates feasibility for using computer-based coding technology within the context of retrospective video analysis methods. Duration of exploratory play was not a discriminating feature of autism at this early age.


Tie1 attenuation reduces murine atherosclerosis in a dose-dependent and shear stress-specific manner.
Although the response of endothelial cells to the disturbed blood flow in the vicinity of atherosclerotic lesions is known to be distinct from that elicited by nonatherogenic laminar flow, the mechanisms involved are poorly understood. Our initial studies confirmed that expression of the endothelial receptor tyrosine kinase Tie1 was evident at regions of atherogenic flow in mature animals. We therefore hypothesized that Tie1 plays a role in the endothelial response to atherogenic shear stress. Consistent with this, we found that Tie1+/- mice bred to the apoE-deficient background displayed a 35% reduction in atherosclerosis relative to Tie1+/+;Apoe-/- mice. Since deletion of Tie1 results in embryonic lethality secondary to vascular dysfunction, we used conditional and inducible mutagenesis to study the effect of endothelial-specific Tie1 attenuation on atherogenesis in Apoe-/- mice and found a dose-dependent decrease in atherosclerotic lesions. Analysis of primary aortic endothelial cells indicated that atheroprotective laminar flow decreased Tie1 expression in vitro. Attenuation of Tie1 was associated with an increase in eNOS expression and Tie2 phosphorylation. In addition, Tie1 attenuation increased IkBα expression while decreasing ICAM levels. In summary, we have found that shear stress conditions that modulate atherogenic events also regulate Tie1 expression. Therefore, Tie1 may play a novel proinflammatory role in atherosclerosis.


Automated localization of cysts in diabetic macular edema using optical coherence tomography images.
This paper presents a novel automated system that localizes cysts in optical coherence tomography (OCT) images of patients with diabetic macular edema (DME). First, in each image, six sub-retinal layers are detected using an iterative high-pass filtering approach. Next, significantly dark regions within the retinal micro-structure are detected as candidate cystoid regions. Each candidate cystoid region is then further analyzed using solidity, mean and maximum pixel value of the negative OCT image as decisive features for estimating the area of cystoid regions. The proposed system achieves 90% correlation between the estimated cystoid area and the manually marked area, and a mean error of 4.6%. Finally the proposed algorithm locates the cysts in the inner plexiform region, inner nuclear region and outer nuclear region with an accuracy of 88%, 86% and 80%, respectively.


Stability and change in symptoms, cognition, and community outcome in schizophrenia.
It has been well established that neurocognitive deficits are a core feature in schizophrenia and predict difficulties in functional independence. However, few studies have assessed the longitudinal stability of cognition and key aspects of functional outcome concurrently. Even less attention has been directed at the contingency of cognitive change on real world outcome changes. Accordingly, this study will assess the extent to which significant changes in cognition and community status are independent or related. As a point of comparison, the stability of clinical symptom status and the relationship between symptom and outcome change are evaluated. Symptoms, cognitive abilities, and community outcome was assessed in 128 patients with schizophrenia at baseline and again one year later. Intraclass correlation coefficients were used to index stability and reliable change index analyses quantified the prevalence of significant improvement or deterioration in each of the three illness features. Results from these analyses revealed that symptom status, cognitive functioning, and community outcome are similarly stable in treated schizophrenia outpatients. A small proportion of the sample demonstrated significant improvement or deterioration in these domains, with only weak evidence that such change was predicted by changes in symptoms or cognition. Further, there was no strong evidence of a preferential relationship for cognition relative to symptoms in relation to functional outcome. These results shed light on the strength and nature of the cognition-real world outcome relationship in schizophrenia and have implications for pharmacological and behavioral interventions aimed at improving real world outcome.


Effects of feedstock ratio and organic loading rate on the anaerobic mesophilic co-digestion of rice straw and cow manure.
In order to investigate the effects of feedstock ratio and organic loading rate (OLR) on the anaerobic mesophilic co-digestion of rice straw (RS) and cow manure (CM), batch tests (2.5L) were carried out at volatile solid (VS) ratios of 0:1, 1:2, 1:1, 2:1, and 1:0 (RS/CM), and continuous bench experiments (40 L) were carried out at OLRs of 3.0, 3.6, 4.2, 4.8, 6.0, 8.0, and 12.0 kg VS/(m(3) d) with optimal VS ratio. The optimal VS ratio was found to be 1:1. Stable and efficient co-digestion with average specific biogas production of 383.5L/kg VS and volumetric biogas production rate of 2.30 m(3)/(m(3) d) was obtained at an OLR of 6 kg VS/(m(3) d). Anaerobic co-digestion was severely inhibited by the accumulation of volatile fatty acids instead of ammonia when the OLR was 12 kg VS/(m(3) d). Further, significant foaming was observed at OLR ⩾ 8 kg VS/(m(3) d).


Progastrin stimulates colonic cell proliferation via CCK2R- and β-arrestin-dependent suppression of BMP2.
Progastrin stimulates colonic mucosal proliferation and carcinogenesis through the cholecystokinin 2 receptor (CCK2R)-partly by increasing the number of colonic progenitor cells. However, little is known about the mechanisms by which progastrin stimulates colonic cell proliferation. We investigated the role of bone morphogenetic proteins (BMPs) in progastrin induction of colonic cell proliferation via CCK2R. We performed microarray analysis to compare changes in gene expression in the colonic mucosa of mice that express a human progastrin transgene, gastrin knockout mice, and C57BL/6 mice (controls); the effects of progastrin were also determined on in vitro colonic crypt cultures from cholecystokinin 2 receptor knockout and wild-type mice. Human colorectal and gastric cancer cells that expressed CCK2R were incubated with progastrin or Bmp2; levels of β-arrestin 1 and 2 were knocked down using small interfering RNAs. Cells were analyzed for progastrin binding, proliferation, changes in gene expression, and symmetric cell division. The BMP pathway was down-regulated in the colons of human progastrin mice compared with controls. Progastrin suppressed transcription of Bmp2 through a pathway that required CCK2R and was mediated by β-arrestin 1 and 2. In mouse colonic epithelial cells, down-regulation of Bmp2 led to decreased phosphorylation of Smads1/5/8 and suppression of inhibitor of DNA binding 4. In human gastric and colorectal cancer cell lines, CCK2R was necessary and sufficient for progastrin binding and induction of proliferation; these effects were blocked when cells were incubated with recombinant Bmp2. Incubation with progastrin increased the number of CD44(+), bromodeoxyuridine+, and NUMB(+) cells, indicating an increase in symmetric divisions of putative cancer stem cells. Progastrin stimulates proliferation in colons of mice and cultured human cells via CCK2R- and β-arrestin 1 and 2-dependent suppression of Bmp2 signaling. This process promotes symmetric cell division.


Respiratory muscle training in chronic obstructive pulmonary disease: inspiratory, expiratory, or both?
Most patients with significant chronic obstructive pulmonary disease (COPD) have inspiratory and expiratory muscle weakness. In addition, hyperinflation induces functional weakening of the inspiratory muscles, increased elastic load to breathing, and intrinsic positive end expiratory pressure (PEEPi). Therefore, it was rational to expect that patients with COPD would benefit from specific inspiratory or expiratory muscle training (SIMT, SEMT respectively). However, the functional benefits of SIMT have remained equivocal. In recent years, a number of studies have demonstrated that, when training loads are controlled, SIMT results in important functional benefits. The role of SEMT is still unclear. Well-controlled SIMT in patients with COPD leads to relief of dyspnea, during both daily activities and during physical activity. This yields increased exercise tolerance, and thus the capacity to walk, improving health related quality of life. We argue that there is now evidence that SIMT is an important addition to pulmonary rehabilitation programs for patients with COPD. Although two recent studies have shown that SEMT also provides a beneficial effect in patients with COPD, this does not appear to be supplementary to the effect to SIMT. Inspiratory and expiratory muscles can be specifically trained yielding improvements in both strength and endurance. The improvement in inspiratory muscle performance is associated with an improvement in the sensation of dyspnea, exercise tolerance, and quality of life. When the expiratory muscles are specifically trained, a significant increase in exercise performance has also been shown. However, there is probably no additional benefit in combining SEMT with SIMT.


A Smac mimetic augments the response of urothelial cancer cells to gemcitabine and cisplatin.
Cisplatin-based chemotherapy is considered the gold standard for patients with advanced bladder cancer. However, despite initial response, many patients will relapse; therefore, novel salvage treatment strategies are desperately needed. Herein, we studied a mechanism based treatment combination using a Smac mimetic with standard chemotherapy. Using a panel of 10 urothelial cancer cell lines, we exposed them to a combination of gemcitabine, cisplatin, and a Smac mimetic. Sensitivity was determined using a DNA fragmentation assay. We determined that three cell lines (UMUC-3, UMUC-13, and RT4v6) were considered sensitive to the combination of gemcitabine and cisplatin and an additional three cell lines were sensitized to gemcitabine and cisplatin with the addition of the Smac mimetic (UMUC-6, UMUC-12, and UMUC-18). We next explored the constitutive expression of selected members of the IAP family (XIAP, cIAP-1, cIAP-2, and Survivin), the BCL family (BCL-2, BCLXL, and BAX) and Smac using gene expression profiling and western blotting. We determined that RNA and protein expression of SMAC, selected members of the IAP family and members of the BCL family did not correlate to drug sensitivity. Lastly, using an in vivo mouse model, we determined that treatment with the Smac mimetic in combination with gemcitabine and cisplatin resulted in increased apoptosis, decreased microvessel density and decreased cellular proliferation. This novel treatment strategy may be effective in patients with advanced urothelial carcinoma and warrants further investigation.


Cervical spine manipulation: an alternative medical procedure with potentially fatal complications.
There are multiple reports in the literature of serious and at times fatal complications after cervical spine manipulation therapy (CSMT), even though CSMT is considered by some health providers to be an effective and safe therapeutic procedure for head and neck pain syndromes. We report a case of a young female with cervicalgia and headache with fatal posterior circulation cerebrovascular accident after CSMT. Serious complications are infrequent, with a reported incidence between one per 100,000 to one in 2 million manipulations. The most frequent injuries involve artery dissection or spasm. Stroke as a complication of cervical manipulation and dissection of the vertebral arteries (VAD) is a rare but well recognized problem. Neck pain, headache, vertigo, vomiting and ataxia are typical symptoms of VAD, but this vascular injury also can be asymptomatic. The most common risk factors are migraine, hypertension, oral contraceptive pills and smoking. Stroke following CSMT is more common than the literature reports. The best values derive from retrospective surveys. The lack of identifiable risk factors place those who undergo CSMT at risk of neurologic damage. Accurate patient information and early recognition of the symptoms are important to avoid catastrophic consequences.


Estimating age-dependent per-encounter chlamydia trachomatis acquisition risk via a Markov-based state-transition model.
Chlamydial infection is a common bacterial sexually transmitted infection worldwide, caused by C. trachomatis. The screening for C. trachomatis has been proven to be successful. However, such success is not fully realized through tailoring the recommended screening strategies for different age groups. This is partly due to the knowledge gap in understanding how the infection is correlated with age. In this paper, we estimate age-dependent risks of acquiring C. trachomatis by adolescent women via unprotected heterosexual acts. We develop a time-varying Markov state-transition model and compute the incidences of chlamydial infection at discrete age points by simulating the state-transition model with candidate per-encounter acquisition risks and sampled numbers of unit-time unprotected coital events at different age points. We solve an optimization problem to identify the age-dependent estimates that offer the closest matches to the observed infection incidences. We also investigate the impact of antimicrobial treatment effectiveness on the parameter estimates and the differences between the acquisition risks for the first-time infections and repeated infections. Our case study supports the beliefs that age is an inverse predictor of C. trachomatis transmission and that protective immunity developed after initial infection is only partial. Our modeling method offers a flexible and expandable platform for investigating STI transmission.


Clinicopathological and immunohistochemical study of 30 cases of post-radiation atypical vascular lesion of the breast.
Atypical vascular lesions (AVL) that occur in the field of prior radiation therapy for breast carcinoma are placed within the differential diagnosis with low grade angiosarcoma and other benign vascular lesions. Although considered a benign entity, the exact biological behavior of AVLs is not fully established because of the small number of cases reported in the literature. We aim to further characterize these lesions clinically and histopathologically, and to study their behavior. We report a series of 30 patients with AVL of the breast occurring after radiation exposure, diagnosed and treated at the European Institute of Oncology, Italy. Immunohistochemical study was performed in all cases, using CD31, D2-40, CD105, and Ki-67 antibodies. Twenty-seven patients were treated with standard doses of conventional adjuvant radiation therapy for the prior breast carcinoma. Three patients were treated with intraoperative radiotherapy with electrons. The post-radiation latency interval from breast carcinoma to AVL was 48.5 months (ranged from 1 to 146 months). Most of the lesions were classified as lymphatic type (78.6 %) based on D2-40 positivity. No extension into subcutaneous tissue or significant atypia was noted in all cases. Despite the fact that the AVL of our series have shown benign behavior in 93.3 %, one patient developed local recurrence of AVL, and two cases progressed to angiosarcoma at the previous AVL site. Further studies should be conducted to better understand the clinical behavior and to propose additional histopathologic diagnostic criteria to distinguish AVL from low grade angiosarcoma and those AVL with increased risk for malignant progression. Concerning current treatments of AVL, we recommend complete excision with free surgical margins and close follow up.


Zolpidem dependence case series: possible neurobiological mechanisms and clinical management.
Zolpidem is a short-acting imidazopyridine hypnotic that is an agonist at the gamma-aminobutyric acid A type (GABAA) receptor. It has been suggested that it acts selectively on alpha1 subunit-containing GABAA benzodiazepine (BZ1) receptors presenting (contrary to classic benzodiazepines) low or no affinity for other subtypes. Therefore, it has been proposed that it lacks the benzodiazepines-like side-effects, having minimal abuse and dependence potential. Nevertheless, there is a considerable number of zolpidem dependence case reports in the literature. We present eight cases of zolpidem abuse and dependence without criminal record, without history of substance abuse (except for one alcohol abuser), with minor psychiatric disorders, who took zolpidem after physicians prescription in order to deal with their insomnia. However, they became zolpidem abusers not craving its sedative, but its anxiolytic and stimulating action, which helped them to cope with everyday activities. It is possible that, in the high doses that our patients used, zolpidem abandons its selectivity for BZ1 receptors and demonstrates all the actions of classic benzodiazepines. Molecular biology, via possible mutations on GABA receptors, may provide some answers as to why our eight patients (who did not differ much from the thousands of insomniacs who use zolpidem) and other zolpidem abusers, raised the dose progressively, and sought something from the drug other than hypnotic action.


Association of elevated levels of C-reactive protein with breast cancer, breast cancer subtypes, and poor outcome.
Inflammation and caner are linked in a bidirectional manner. C-reactive protein (CRP) is an important inflammatory marker. The aim of the study was to test whether the inflammatory marker, CRP at the time of diagnosis of breast cancer is associated with metastasis, recurrence, and death in breast cancer patients from Malwa region of Punjab where breast cancer is widely feared. Two hundred and forty-two breast cancer patients and 242 age and sex matched controls were included in the study. CRP levels were estimated using fully automated bio analyzer Erba200. Follow up interviews were conducted at an interval of 3, 6, 9, 12, 15, 18, 21, 24, and 27 months to determine the outcome among breast cancer patients. Elevated levels of CRP were found among the diseased in comparison with controls (P < 0.0001). Higher CRP levels associated significantly with poor outcome including metastasis and recurrence among breast cancer patients [P = 0.03; 95% confidence interval; odds ratio: 2.954 (0.9125-9.561)]. Elevated levels of CRP associated significantly with increased risk of breast cancer and poor outcome. CRP estimation may be a simple and inexpensive tool for the risk assessment and outcome of the disease in Malwa region of Punjab where incidence of breast cancer is reported to be very high.


Structural insights into targeting of the colchicine binding site by ELR510444 and parbendazole to achieve rational drug design.
Microtubules consisting of α- and β-tubulin heterodimers have proven to be an efficient drug target for cancer therapy. A broad range of agents, including ELR510444 and parbendazole, can bind to tubulin and interfere with microtubule assembly. ELR510444 and parbendazole are colchicine binding site inhibitors with antiproliferative activities. However, the lack of structural information on the tubulin-ELR510444/parbendazole complex has hindered the design and development of more potent drugs with similar scaffolds. Therefore, we report the crystal structures of tubulin complexed with ELR510444 at a resolution of 3.1 Å and with parbendazole at 2.4 Å. The structure of these complexes revealed the intermolecular interactions between the two colchicine binding site inhibitors and tubulin, thus providing a rationale for the development of novel benzsulfamide and benzimidazole derivatives targeting the colchicine binding site.


ZC3H4 governs epithelial cell migration through ROCK/p-PYK2/p-MLC2 pathway in silica-induced pulmonary fibrosis.
Increased epithelial migration capacity is a key step accompanying epithelial-mesenchymal transition (EMT). Our lab has described that ZC3H4 mediated EMT in silicosis. Here, we aimed to explore the mechanisms of ZC3H4 by which to stimulate epithelial cell migration. Silicon dioxide (SiO2)-induced pulmonary fibrosis (PF) animal models were administered by intratracheal instillation in C57BL/6 J mice. Pathological analysis and 2D migration assay were established to uncover the pulmonary fibrotic lesions and epithelial cell migration, respectively. Inhibitors targeting ROCK/p-PYK2/p-MLC2 and CRISPR/Cas9 plasmids targeting ZC3H4 were administrated to explore the signaling pathways. 1) SiO2 upregulated epithelial migration in pulmonary fibrotic lesions. 2) ZC3H4 modulated SiO2-induced epithelial migration. 3) ZC3H4 governed epithelial migration through ROCK/p-PYK2/p-MLC2 signaling pathway. ZC3H4 regulates epithelial migration through the ROCK/p-PYK2/p-MLC2 signaling pathway, providing the possibility that molecular drugs targeting ZC3H4-overexpression may exert effects on pulmonary fibrosis induced by silica.


[The role of image charge in ionic-electrostatic interactions of proteins and membranes].
The interaction of ions of electrolyte cytoplasm with the electric fields of image charges in adjacent dielectrics (membranes, proteins) was computed. The screening effect of neighbouring ions on the electric field of each image charge was taken into account. It is shown that the forces produced by these image charges are significant only at distances from dielectric surfaces that do not exceed three-four average distances between the charges in the cytoplasm. The forces and energies were computed with which liquid ions push away the surfaces of two adjacent membranes or proteins. It is shown that these values exceed hundreds times normal ionic-electrostatic interactions. The proposed approach taken it possible to explain the experimentally observed behaviour of on cell adhesion, the collective mode of protein interaction in plasma membranes and other phenomena.


Reduced stress concentration and enhanced fracture toughness by yielding-rehardening combination.
A range of tough materials (e.g., metals and polymer solids) exhibit a characteristic mechanical behavior, that is, a combination of yielding and subsequent rehardening. We numerically investigate how the combination of these mechanical behaviors enhances resistance to crack propagation.Our system has a 2-dimensional square lattice structure where pairs of adjacent lattice points are connected by "special" bonds. An isolated bond behaves as a linear spring for small deformations, but yields at a threshold force to produce a plateau in its force-deformation curve, and then shows rehardening on further loading up to a critical force of bond-breaking; on unloading from above the yielding point, the force rapidly decreases with deformation (hysteresis). We simulate crack propagation in the entire system (the square lattice structure) from an initial crack driven by boundary loading. The threshold force for bond-yielding is varied as a simulation parameter, while the critical force for bond-breaking in the rehardening regime is fixed to 1. In other words, the substantial simulation parameter is the ratio between the yielding and breaking forces. We find that the fracture behavior drastically changes depending on the ratio: when the ratio is low, the bond-breaking energy (of a single bond) is low, but more work is required to fracture the entire system via the crack propagation. The opposite tendency between the bond-breaking energy and the fracture work is due to formation of a well-developed yielding zone around the crack tip.


Phase II trial of carboxyamidotriazole in patients with relapsed epithelial ovarian cancer.
Carboxyamidotriazole (CAI) is a cytostatic inhibitor of nonvoltage-operated calcium channels and calcium channel-mediated signaling pathways. It inhibits angiogenesis, tumor growth, invasion, and metastasis. We hypothesized that CAI would promote disease stabilization lasting >/= 6 months in patients with relapsed ovarian cancer. Patients with epithelial ovarian cancer, good end-organ function, measurable disease, and three or fewer prior regimens were eligible. Oral CAI was given daily using a pharmacokinetic-dosing approach to maintain plasma concentrations between 2 and 4 microg/mL. Radiographic imaging to assess response was performed every 8 weeks. Positive outcome included stabilization or improvement of disease lasting >/= 6 months. Plasma vascular endothelial growth factor (VEGF), interleukin (IL)-8, and matrix metalloproteinase (MMP)-2 were measured. Thirty-six patients were assessable for primary end point analysis, and 38 were assessable for toxicity. Forty-four percent of patients had three prior regimens, more than 50% had four or more disease sites, and 48% had liver metastases. Thirty-three patients reached the targeted concentration range during the first cycle. Eleven patients (31%) attained the >/= 6-month outcome end point, with one partial response (8 months) and three minor responses (8, 12+, and 13 months). Median time to progression was 3.6 months (range, 1.6 to 13.3 months). CAI was well tolerated, with mostly grade 1 to 2 toxicity. Grade 3 events included fatigue (5%), vomiting (2%), neutropenic fever (2%), and neutropenia (2%). There were no grade 4 adverse events. No associations between VEGF, IL-8, and MMP-2 with CAI concentration or clinical outcome were observed. CAI is a potential agent for additional study in the stabilization of relapsed ovarian cancer. Given a limited toxicity profile, it may have utility as a maintenance therapeutic agent for this disease.


Peripheral synapses at identified mechanosensory neurons in spiders: three-dimensional reconstruction and GABA immunocytochemistry.
The mechanosensory organs of arachnids receive diverse peripheral inputs. Little is known about the origin, distribution, and function of these chemical synapses, which we examined in lyriform slit sense organ VS-3 of the spider Cupiennius salei. The cuticular slits of this organ are each associated with two large bipolar mechanosensory neurons with different adaptation rates. With intracellular recording, we have now been able to correlate directly the staining intensity of a neuron for acetylcholinesterase with its adaptation rate, thus allowing us simply to stain a neuron to identify its functional type. All rapidly adapting neurons stain more heavily than slowly adapting neurons. Immunostaining of whole-mount preparations reveals GABA-like immunoreactive fibers forming numerous varicosities at the surface of all sensory neurons in VS-3; peripheral GABA-like immunoreactive somata are lacking. Sectioning the leg nerve procures rapid degeneration of most fiber profiles, confirming that the fibers are efferent. Punctate synapsin-like immunoreactivity colocalizes to these varicosities, although some synapsin-like immunoreactive puncta are GABA-immunonegative. Fibers with similar immunoreactivities are also associated with trichobothria, tactile hairs, internal joint receptors, i.e. other types of spider mechanosensory organs. In organ VS-3, immunoreactivity is most dense across the initial axon segment. The exact distribution of peripheral synapses was reconstructed from a 10-microm-long electron micrograph series of the dendritic, somatic, and initial axon regions of acetylcholinesterase-stained VS-3 neurons. These reveal a pattern similar to that of the synapsin-like immunoreactivity. Two different types of synapse were distinguished on the basis of their presynaptic vesicle populations. Many peripheral synapses thus appear to derive from efferent GABA-like immunoreactive fibers and probably provide centrifugal inhibitory control of primary mechanosensory activities.


[Dynamics of blood antioxidative activity in relation to ECG changes and the course of acute myocardial infarct].
Blood serum antioxidation activity (AOA) was studied by the biophysical method of inhibited electrochemiluminescence in 500 patients with acute myocardial infarction. The degree of increased AOA depends on the size of the affected area. AOA intensity is greater in macrofocal than in microfocal myocardial infarction. It was established that in non-complicated myocardial infarction AOA is characterized by smooth dynamics with normalization of the values on the 3d week. Rapid AOA dynamics are encountered in II or III degree chronic congestive circulatory insufficiency. Protracted dynamics of antioxidation activity (longer than 3 weeks) is evidence of a recurrent character of the disease.


Diet-induced over-expression of flightless-I protein and its relation to flightlessness in Mediterranean fruit fly, Ceratitis capitata.
The Mediterranean fruit fly (medfly), Ceratitis capitata is among the most economically important pests worldwide. Understanding nutritional requirement helps rearing healthy medfly for biocontrol of its population in fields. Flight ability is a high priority criterion. Two groups of medfly larvae were reared with two identical component diets except one with fatty acids (diet A) and another without it (diet B). Adults from larvae reared on diet B demonstrated 20±8% of normal flight ability, whereas those from larvae reared on diet A displayed full flight ability of 97±1%. Proteomes were profiled to compare two groups of medfly pupae using shotgun proteomics to study dietary effects on flight ability. When proteins detected in pupae A were compared with those in pupae B, 233 and 239 proteins were, respectively, under- and over-expressed in pupae B, while 167 proteins were overlapped in both pupae A and B. Differential protein profiles indicate that nutritional deficiency induced over-expression of flightless-I protein (fli-I) in medfly. All proteins were subjected to Ingenuity Pathway Analysis (IPA) to create 13 biological networks and 17 pathways of interacting protein clusters in human ortholog. Fli-I, leucine-rich repeat (LRR)-containing G protein-coupled receptor 2, LRR protein soc-2 and protein wings apart-like were over-expressed in pupae B. Inositol-1,4,5-trisphosphate receptor, protocadherin-like wing polarity protein stan and several Wnt pathway proteins were under-expressed in pupae B. These results suggest down-regulation of the Wnt/wingless signaling pathway, which consequently may result in flightlessness in pupae B. The fli-I gene is known to be located within the Smith-Magenis syndrome (SMS) region on chromosome 17, and thus, we speculate that nutritional deficiency might induce over-expression of fli-I (or fli-I gene) and be associated with human SMS. However, more evidence would be needed to confirm our speculation.


Auxeticity enhancement due to size polydispersity in fcc crystals of hard-core repulsive Yukawa particles.
The Poisson's ratio of the fcc hard-core repulsive Yukawa crystals with size polydispersity was determined by Monte Carlo simulations in the isothermal-isobaric ensemble. The effect of size polydispersity on the auxetic properties of Yukawa crystals has been studied. It has been found that an increase of particle size polydispersity causes a decrease of the Poisson's ratio in auxetic directions as well as appearance of a negative Poisson's ratio in formerly non-auxetic directions. A measure of auxeticity was introduced to estimate quantitatively an enhancement of auxetic properties in polydisperse Yukawa crystals. The proposed measure of auxeticity can be applied to appraise the auxeticity of any studied system.


[An approach to the phenomenological interview].
The purpose of this study is to describe my experiences about the phenomenological interview. It was developed in four moments, in order to adequate physical environment aspects, strategies of approaching the interviewers, the orientation question and the techniques to obtain the statements. The focus on researcher-subject/researched-subject has indicated adjustments in the utilization of the techniques and reoriented my person and professional development.


Enhanced eryptosis following gramicidin exposure.
The peptide antibiotic and ionophore gramicidin has previously been shown to trigger apoptosis of nucleated cells. In analogy to apoptosis, the suicidal death of erythrocytes or eryptosis involves cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Triggers of eryptosis include oxidative stress, increase of cytosolic Ca2+ activity ([Ca2+]i), and ceramide. The present study explored, whether gramicidin triggers eryptosis. To this end phosphatidylserine exposure at the cell surface was estimated from annexin V binding, cell volume from forward scatter, red blood cell distribution width (RDW) from electronic particle counting, reactive oxidant species (ROS) from 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) fluorescence, [Ca2+]i from Fluo3- and Fluo4 fluorescence, and ceramide abundance from binding of specific antibodies. As a result, a 24 h exposure of human erythrocytes to gramicidin significantly increased the percentage of annexin-V-binding cells (≥1 µg/mL), forward scatter (≥0.5 µg/mL) and hemolysis. Gramicidin enhanced ROS activity, [Ca2+]i and ceramide abundance at the erythrocyte surface. The stimulation of annexin-V-binding by gramicidin was significantly blunted but not abolished by removal of extracellular Ca2+. In conclusion, gramicidin stimulates phospholipid scrambling of the erythrocyte cell membrane, an effect at least partially due to induction of oxidative stress, increase of [Ca2+]i and up-regulation of ceramide abundance. Despite increase of [Ca2+]i, gramicidin increases cell volume and slightly reduces RWD.


Construction and preliminary characterization of three hemagglutinin mutants of Porphyromonas gingivalis.
Targeted insertional mutagenesis was used to construct hagA, hagB, and hagC hemagglutinin mutants of Porphyromonas gingivalis. pJRD215-derived plasmids containing tetA(Q)2 and portions of the targeted genes were conjugated into P. gingivalis. Interruption of the three loci was confirmed by Southern hybridization, sequencing, reverse transcription-PCR, and microtiter hemagglutination assays. No significant differences in hydrophobicity or coadherence to Actinomyces viscosus were detected between the mutants and the wild-type strain.


The Contributions of Neonatal Inhalation of Copper to Air Pollution-Induced Neurodevelopmental Outcomes in Mice.
Exposures to ambient ultrafine particle (UFP) air pollution (AP) during the early postnatal period in mice (equivalent to human third trimester brain development) produce male-biased changes in brain structure, including ventriculomegaly, reduced brain myelination, alterations in neurotransmitters and glial activation, as well as impulsive-like behavioral characteristics, all of which are also features characteristic of male-biased neurodevelopmental disorders (NDDs). The purpose of this study was to ascertain the extent to which inhaled Cu, a common contaminant of AP that is also dysregulated across multiple NDDs, might contribute to these phenotypes. For this purpose, C57BL/6J mice were exposed from postnatal days 4-7 and 10-13 for 4hr/day to inhaled copper oxide (CuxOy) nanoparticles at an environmentally relevant concentration averaging 171.9ng/m3. Changes in brain metal homeostasis and neurotransmitter levels were determined following termination of exposure (postnatal day 14), while behavioral changes were assessed in adulthood. CuxOy inhalation modified cortical metal homeostasis and produced male-biased disruption of striatal neurotransmitters, with marked increases in dopaminergic function, as well as excitatory/inhibitory imbalance and reductions in serotonergic function. Impulsive-like behaviors in a fixed ratio (FR) waiting-for-reward schedule and a fixed interval (FI) schedule of food reward occurred in both sexes, but more prominently in males, effects which could not be attributed to altered locomotor activity or short-term memory. Inhaled Cu as from AP exposures, at environmentally relevant levels experienced during development, may contribute to impaired brain function, as shown by its ability to disrupt brain metal homeostasis and striatal neurotransmission. In addition, its ability to evoke impulsive-like behavior, particularly in male offspring, may be related to striatal dopaminergic dysfunction that is known to mediate such behaviors. As such, regulation of air Cu levels may be protective of public health.


Lateral habenula cannabinoid CB1 receptor involvement in drug-associated impulsive behavior.
Animal and human studies show that cannabis or its derivatives can increase relapse to cocaine seeking following withdrawal. Moreover, cannabis use in humans is associated with impulse control deficits and animal studies implicate endogenous cannabinoids (eCB) in several impulsivity constructs. However, the brain areas where cannabinoids might control impulsivity or cocaine seeking are largely unknown. Here, we assess Lateral Habenula (LHb) involvement on performance in the 5-choice serial reaction time task (5CSRTT) in rats and investigate whether LHb cannabinoid CB1 receptors (CB1R) are involved in these effects. Systemic cocaine increased premature responding, a measure of impulsivity, at a dose (5 mg/kg) that did not alter other measures of task performance. Intra-LHb infusion of the CB1R antagonist AM251 blocked this effect. Systemic injection of the psychoactive constituent of cannabis, Δ9-tetrahydrocannabinol (Δ9-THC, 1 mg/kg), also increased 5CSRTT premature responding at a dose that did not otherwise disrupt task performance. This was blocked by intra-LHb infusion of AM251 in a subgroup of rats showing the largest increases in Δ9-THC-evoked premature responses. Systemic Δ9-THC also prompted impulsive cocaine seeking in a Go/NoGo cocaine self-administration task and this was blocked by intra-LHb AM251. These data show that LHb CB1Rs are involved in deficits in impulse control initiated by cocaine and Δ9-THC, as assessed by the 5CSRTT, and play a role in impulsive cocaine seeking during cocaine self-administration. This suggests that the LHb eCB system contributes to the control of impulsive behavior, and thus represents a potential target for therapeutic treatment of substance use disorders (SUDs) in humans.


Differential regulation of serum- and glucocorticoid-inducible kinase 1 (SGK1) splice variants based on alternative initiation of transcription.
The serum- and glucocorticoid-inducible kinase 1 (SGK1) is a key-regulator of transport, cell volume and cell survival. SGK1 transcription is under genomic control of a wide variety of hormones and cell stressors. Little is known, however, about sequence variation in SGK1 transcripts. Thus, we took an in silico approach to determine sequence variations in the N-terminal region of SGK1, which is considered particularly important for subcellular SGK1 localization. Expressed Sequence Tag analysis revealed two novel phylogenetically highly conserved SGK1 mRNAs with different promoter sites based on alternative initiation of transcription at -2981, -850 upstream of the transcription initiation site (+1) of the reference mRNA. RT-PCR in various human cell lines and tissues confirmed the expression of the 3 alternative splice variants, which differed exclusively in their first exons. The two novel variants were devoid of the localization and degradation signal with otherwise unchanged and intact open reading frames. Spatial distribution of transcription factor binding sites among the three promoter sites indicated common responsiveness to glucocorticoids but different responsiveness to hypoxia and cellular differentiation. Differential expression under those conditions was confirmed for all variants in cultured myoblasts and myotubes. p53 and ETF-1 binding sites were overrepresented at the promoter site of the reference sequence variant SGK1(+1). Transcript levels were 4.1-fold [SGK1(+1)] and 3.1-fold [SGK1(-850)] higher in renal clear cell carcinoma than in remote tissue. The transcript levels were 42-fold [SGK1(+1)], 26-fold [SGK1(-850)] and 17-fold [SGK1(-2981)] higher in highly malignant human glioma cells than in non-neoplastic brain tissue. SGK1 transcript levels were differentially increased by differentiation or hypoxia (treatment with CoCl(2)). In conclusion, the present observations disclose the transcription of three distinct SGK1 splice variants, which are all markedly upregulated in tumor tissue but differentially upregulated following differentiation or hypoxia.


[Effects of atmospheric calibration on remote sensing estimation of Moso bamboo forest biomass].
Landsat Thematic Mapper (TM) image was used to estimate Moso bamboo forest biomass, and six atmospheric calibration methods (FLAASH model, 6S model, and DOS1-4 models) were adopted to analysis the effects of atmospheric calibration on the remote sensing estimation of Moso bamboo forest biomass. All the six calibration methods could effectively reduce the atmospheric impacts on TM spectral responses. The relationships between NDVI and Moso bamboo forest biomass under the calibration by the six calibration methods were improved. Great differences were observed in the relationships of Moso bamboo forest biomass with NDVI, II, and MI when using the same calibration methods, suggesting that atmospheric calibration should be made for studying the biophysical significance of vegetation indices. The Landsat TM data corrected with DOS3 model had the highest correlation coefficient with Moso bamboo forest biomass, but there were no significant differences in the correlation coefficients after corrected with the six calibration methods, which indicated that atmospheric calibration might be not required if a single TM image was used for biomass estimation with multiple linear regression model.


The influence of surgical specialty and caseload on the results of carotid endarterectomy.
Carotid endarterectomy is rapidly becoming one of the most commonly performed major surgical operations in the United States, in part because of the greater availability of noninvasive techniques to accurately diagnose extracranial carotid arterial disease and a low reported morbidity and mortality. We retrospectively reviewed the records for all carotid endarterectomies performed in the greater Cincinnati area for a recent 12-month period and examined the impact of surgical specialty and operative caseload on the results. Altogether, 750 operations were performed on 656 patients by 61 surgeons working in 16 general medical-surgical hospitals. Overall, strokes occurred in 5.1% of all patients; 2.3% of patients died. Symptomatic patients had a significantly higher risk of suffering a postoperative stroke compared with asymptomatic patients (6.5% vs. 3.7%), although the risk of death was virtually identical (2.4% vs. 2.1%). When the operating surgeons were classified into four types on the basis of their previous training, no statistically significant differences in either postoperative stroke or death could be identified. Furthermore, when the surgical caseloads of these physicians were grouped into three categories (i.e., less than 12 each year, more than 50 each year, and a group between these two extremes), no significant differences in outcome were seen. We concluded that our community-wide results for carotid endarterectomy were not comparable to those previously published from specialized centers and that these results did not appear to be influenced by the type of formal surgical specialty or operative caseload.


Obesity and non-alcoholic fatty liver disease in chronic hepatitis C.
Superimposed non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) may affect HCV-related fibrosis. We performed a study to determine the relationship between NAFLD and chronic hepatitis C. One hundred and twenty patients with chronic hepatitis C and available liver biopsies were included. Baseline liver biopsies were read by 1 hepatopathologist using Metavir, as well as a fatty liver pathology protocol. Patients' baseline clinical, demographic, and virologic data were associated with the extent of steatosis (>33% vs. < or =33%), the type of fatty liver (no steatosis vs. steatosis only vs. NASH), and the stage of fibrosis seen on the liver biopsy. Seventy percent of patients were men and 80% were white. The mean age was 47.48+/-5.70 years, mean BMI was 29.01 +/-5.01 kg/m, and mean waist to hip ratio (W/H) was 0.90+/-0.08. Patients with higher grade of steatosis had higher BMI (32.83+/-6.26 vs. 28.49+/-4.62, P = 0.034), more likely to have genotype 3 (21.4% vs. 5.7%, P = 0.037) and advanced fibrosis (92.9% vs. 62.3%, P = 0.033) than those with lower grade of steatosis. Of these, only HCV-genotype 3 remained independently associated with higher grade of steatosis. When patients with superimposed NASH (n = 22) were compared with those with only steatosis (n = 49) and those without steatosis (n = 49), patients with superimposed NASH had more evidence of obesity (BMI: 30.64+/-5.23 vs. 29.90+/-5.35 vs. 27.33+/-4.07, P = 0.008; W/H: 0.97+/-0.06 vs. 0.91+/-0.08 vs. 0.87+/-0.07, P < 0.001), more commonly infected with HCV genotype 3 (14% vs. 12% vs. 0%, P = 0.036) and had more advanced fibrosis (95.5% vs. 75.5% vs. 42.9%, P < 0.001). Race, gender, and age did not affect extent of steatosis or presence of superimposed NASH. In conclusion, markers of obesity (BMI and W/H) and HCV genotype 3 are associated with the extent of steatosis and type of fatty liver. Higher grade of steatosis and presence of superimposed NASH are both associated with advanced hepatic fibrosis.


Assessing the efficacy of hearing-aid amplification using a phoneme test.
Consonant-vowel (CV) perception experiments provide valuable insights into how humans process speech. Here, two CV identification experiments were conducted in a group of hearing-impaired (HI) listeners, using 14 consonants followed by the vowel /ɑ/. The CVs were presented in quiet and with added speech-shaped noise at signal-to-noise ratios of 0, 6, and 12 dB. The HI listeners were provided with two different amplification schemes for the CVs. In the first experiment, a frequency-independent amplification (flat-gain) was provided and the CVs were presented at the most-comfortable loudness level. In the second experiment, a frequency-dependent prescriptive gain was provided. The CV identification results showed that, while the average recognition error score obtained with the frequency-dependent amplification was lower than that obtained with the flat-gain, the main confusions made by the listeners on a token basis remained the same in a majority of the cases. An entropy measure and an angular distance measure were proposed to assess the highly individual effects of the frequency-dependent gain on the consonant confusions in the HI listeners. The results suggest that the proposed measures, in combination with a well-controlled phoneme speech test, may be used to assess the impact of hearing-aid signal processing on speech intelligibility.


Bioengineered Nanoparticles Loaded-Hydrogels to Target TNF Alpha in Inflammatory Diseases.
Rheumatoid Arthritis (RA) is an incurable autoimmune disease that promotes the chronic impairment of patients' mobility. For this reason, it is vital to develop therapies that target early inflammatory symptoms and act before permanent articular damage. The present study offers two novel therapies based in advanced drug delivery systems for RA treatment: encapsulated chondroitin sulfate modified poly(amidoamine) dendrimer nanoparticles (NPs) covalently bonded to monoclonal anti-TNF α antibody in both Tyramine-Gellan Gum and Tyramine-Gellan Gum/Silk Fibroin hydrogels. Using pro-inflammatory THP-1 (i.e., human monocytic cell line), the therapy was tested in an inflammation in vitro model under both static and dynamic conditions. Firstly, we demonstrated effective NP-antibody functionalization and TNF-α capture. Upon encapsulation, the NPs were released steadily over 21 days. Moreover, in static conditions, the approaches presented good anti-inflammatory activity over time, enabling the retainment of a high percentage of TNF α. To mimic the physiological conditions of the human body, the hydrogels were evaluated in a dual-chamber bioreactor. Dynamic in vitro studies showed absent cytotoxicity in THP-1 cells and a significant reduction of TNF-α in suspension over 14 days for both hydrogels. Thus, the developed approach showed potential for use as personalized medicine to obtain better therapeutic outcomes and decreased adverse effects.


Vitamin D Inhibits IL-22 Production Through a Repressive Vitamin D Response Element in the il22 Promoter.
Th22 cells constitute a recently described CD4+ T cell subset defined by its production of interleukin (IL)-22. The action of IL-22 is mainly restricted to epithelial cells. IL-22 enhances keratinocyte proliferation but inhibits their differentiation and maturation. Dysregulated IL-22 production has been associated to some inflammatory skin diseases such as atopic dermatitis and psoriasis. How IL-22 production is regulated in human T cells is not fully known. In the present study, we identified conditions to generate Th22 cells that do not co-produce IL-17 from naïve human CD4+ T cells. We show that in addition to the transcription factors AhR and RORγt, the active form of vitamin D3 (1,25(OH)2D3) regulates IL-22 production in these cells. By studying T cells with a mutated vitamin D receptor (VDR), we demonstrate that the 1,25(OH)2D3-induced inhibition of il22 gene transcription is dependent on the transcriptional activity of the VDR in the T cells. Finally, we identified a vitamin D response element (VDRE) in the il22 promoter and demonstrate that 1,25(OH)2D3-VDR directly inhibits IL-22 production via this repressive VDRE.


Association of autoimmunity to peptidyl arginine deiminase type 4 with genotype and disease severity in rheumatoid arthritis.
Protein citrullination is an important posttranslational modification recognized by rheumatoid arthritis (RA)-specific autoantibodies. One of the citrullinating enzymes, peptidyl arginine deiminase type 4 (PAD-4), is genetically associated with development of RA in some populations, although the mechanism(s) mediating this effect are not yet clear. There have been descriptions of anti-PAD-4 autoantibodies in different rheumatic diseases. This study was undertaken to investigate whether anti-PAD-4 antibodies are specific to RA, are associated with disease phenotype or severity, and whether PAD-4 polymorphisms influence the anti-PAD-4 autoantibody response. Sera from patients with established RA, patients with other rheumatic diseases, and healthy adults were assayed for anti-PAD-4 autoantibodies by immunoprecipitation of in vitro-translated PAD-4. The epitope(s) recognized by PAD-4 autoantibodies were mapped using various PAD-4 truncations. PAD-4 genotyping was performed on RA patients with the TaqMan assay. Joint erosions were scored from hand and foot radiographs using the Sharp/van der Heijde method. PAD-4 autoantibodies were found in 36-42% of RA patients, and were very infrequent in controls. Recognition by anti-PAD-4 autoantibodies required the 119 N-terminal amino acids, which encompass the 3 nonsynonymous polymorphisms associated with disease susceptibility. Strikingly, the anti-PAD-4 immune response was associated with the RA susceptibility haplotype of PADI4. Anti-PAD-4 antibodies were associated with more severe joint destruction in RA. Our findings indicate that anti-PAD-4 antibodies are specific markers of RA, independently associated with more severe disease, suggesting that an anti-PAD-4 immune response may be involved in pathways of joint damage in this disease. Polymorphisms in the PADI4 gene influence the immune response to the PAD-4 protein, potentially contributing to disease propagation.


Pillar arene Se nanozyme therapeutic systems with dual drive power effectively penetrated mucus layer combined therapy acute lung injury.
siRNA has demonstrated a promising paradigm for therapy of acute lung injury(ALI). However, the pulmonary mucus layer barrier powerfully hinders the therapeutic efficacy. Herein, we proposed to use dual drive power to enhance the mucus permeation of siRNA by constructing the neutral and targeted selenium nanozymes therapeutic system. The multifunctional selenium nanozymes (CWP-Se@Man) were synthesized by modifying with cationic water-soluble pillar arene (CWP) and mannose (Man). After loading CCR2-siRNA, the CWP-Se@Man reached electroneutrality that co-driven by electroneutrality and targeting, the mucus permeation capacity of CWP-Se@Man enhanced by ∼15 fold, thus effectively penetrate pulmonary mucus layer and deliver CCR2-siRNA into macrophages. Moreover, with optimizing the composition of CWP-Se@Man made of CWP (Slutsky, 2013) [5] or CWP (Ichikado et al., 2012) [6], the therapeutic system CWP (Ichikado et al., 2012) [6]-Se@Man showed better biological activities due to smaller size. In inflamed modes, the CWP-Se@Man nanotherapeutic systems loading CCR2-siRNA not only exerted pronounced anti-inflammatory effect through combining inhibit the chemotactic effect and ROS, but also effectively against ALI after blocking the circulatory effect of ROS and inflammatory cytokines. Therefore, this strategy of dual-driving force penetration mucus renders a unique approach for mediating trans-mucus nucleic acid delivery in lungs, and provide a promising treatment for the acute lung injury therapy.


Double-grooved nanofibre surfaces with enhanced anisotropic hydrophobicity.
This study reports a facile method for fabricating double-grooved fibrous surfaces. The primary grooves of the surface are formed by aligned fibres, while the secondary grooves are achieved by oriented nanogrooves on the fibre surface. Investigation into the formation mechanism reveals that the nanogrooves can be readily tailored through adjusting the solvent ratio and relative humidity. With this understanding, a variety of polymers have been successfully electrospun into fibres having the same nanogrooved feature. These fibres show high resemblance to natural hierarchical structures, and thereby endowing the corresponding double-grooved surface with enhanced anisotropic hydrophobicity. A water droplet at a parallel direction to the grooves exhibits a much higher contact angle and a lower roll-off angle than the droplet at a perpendicular direction. The application potential of such anisotropic hydrophobicity has been demonstrated via a fog collection experiment, in which the double-grooved surface can harvest the largest amount of water. Moreover, the fabrication method requires neither post-treatment nor sophisticated equipment, making us anticipate that the double-grooved surface would be competitive in areas where a highly ordered surface, a large surface area and an anisotropic hydrophobicity are preferred.


Child behavioural problems and body size among 2-6 year old children predisposed to overweight. results from the "healthy start" study.
Psychological adversities among young children may be associated with childhood overweight and obesity. We examined if an increased level of child behavioural problems was associated with body size among a selected group of 2-6 year old children, who were all predisposed to develop overweight. Cross-sectional analyses were conducted using baseline data from the "Healthy Start" intervention study. A total of 3058 children were invited to participate, and data from 583 children who were all predisposed for obesity was analyzed. The Danish version of the Strengths and Difficulties Questionnaire (SDQ) was used to assess child stress by the SDQ Total Difficulties (SDQ-TD) score and the Prosocial Behavior (PSB) score. Height and weight were measured, and BMI z-scores were calculated. A direct, but non-significant linear trend was found between SDQ-TD score and BMI z-score (β = 0.021, p = 0.11). Having an SDQ-TD score above the 90(th) percentile was associated with BMI z-score (β = 0.36, p = 0.05). PSB score was not associated with BMI z-score. Analyses were adjusted for parental socioeconomic status, parental BMI, family structure, dietary factors, physical activity, and family stress level. The results suggested a threshold effect between SDQ-TD score and BMI z-score, where BMI z-score was associated with childhood behavioural problems only for those with the highest scores of SDQ-TD. No significant association between PSB score and BMI z-score was found.


Milk conjugated linoleic acid response to fish oil supplementation of diets differing in fatty acid profiles.
The objective of this experiment was to examine the effect of feeding fish oil (FO) along with fat sources that varied in their fatty acid compositions (high stearic, high oleic, high linoleic, or high linolenic acids) to determine which combination would lead to maximum conjugated linoleic acid (cis-9,trans-11 CLA) and transvaccenic acid (TVA) concentrations in milk fat. Twelve Holstein cows (eight multiparous and four primiparous cows) at 73 (+/- 32) DIM were used in a 4 x 4 Latin square with 4-wk periods. Treatment diets were 1) 1% FO plus 2% fat source high in stearic acid (HS), 2) 1% FO plus 2% fat from high oleic acid sunflower seeds (HO), 3) 1% FO plus 2% fat from high linoleic acid sunflower seeds (HLO), and 4) 1% FO plus 2% fat from flax seeds (high linolenic; HLN). Diets formulated to contain 18% crude protein were composed of 50% (dry basis) concentrate mix, 25% corn silage, 12.5% alfalfa haylage, and 12.5% alfalfa hay. Milk production (35.8, 36.3, 34.9, and 35.0 kg/d for diets 1 to 4) was similar for all diets. Milk fat percentages (3.14, 2.81, 2.66, and 3.08) and yields (1.13, 1.02, 0.93, and 1.08 kg/d) for diets 1 to 4 were lowest for HLO. Milk protein percentages (3.04, 3.03, 3.10, and 3.08) and dry matter intake (DMI) (25.8, 26.0, 26.2, and 26.2 kg/d) for diets 1 to 4 were similar for all diets. Milk cis-9,trans-11 CLA concentrations (0.70, 1.04, 1.70, and 1.06 g/100 g fatty acids) for diet 1 to 4 and yields (7.7, 10.7, 15.8, and 11.3 g/d) for diets 1 to 4 were greatest with HLO and were least with HS. Milk cis-9,trans-11 CLA concentrations and yields were similar for cows fed the HO and the HLN diets. Similar to milk cis-9,trans-11 CLA, milk TVA concentration (1.64, 2.49, 3.74, and 2.41 g/100 g fatty acids) for diets 1 to 4 was greatest with the HLO diet and least with the HS diet. Feeding a high linoleic acid fat source with fish oil most effectively increased concentrations and yields of milk cis-9,trans-11 CLA and TVA.


Comparison of thermal coagulation profiles for bipolar forceps with different cooling mechanisms in a porcine model of spinal surgery.
Coagulation accomplished using bipolar forceps is common in neurosurgery. Control of thermal spread from the forceps tips into surrounding neural tissues is a persistent concern, as neural tissues are especially vulnerable to heat injury. The purpose of our investigation was to compare the efficacy of cooling mechanisms for four different bipolar forceps and to understand thermal spread when coagulating vessels on the spinal cord. Immediately following euthanasia, the dura mater of an ex vivo porcine model was opened to expose vessels on the spinal cord for coagulation. Temperature profiles were measured at generator power of 25 W and at fixed 5-second activation times. The bipolar forceps used in this study included regular stainless steel, titanium, heat-pipe embedded, and SILVERGlide forceps. Temperature was measured by micro-thermistor at the midpoint between the bipolar tips, and 1 and 2 mm away from the midpoint along the centerline. Statistical analysis was performed to evaluate temperature differences. Temperature profiles indicated that heat-pipe embedded forceps create the least amount of temperature increase and the highest normalized temperature decreasing slope after activation. The decreasing slope of SILVERGlide forceps is slightly higher than that of regular stainless steel forceps. Bipolar forceps incorporating either heat-pipe embedded technology or SILVERGlide coating can effectively limit excessive thermal spread, thus decreasing potential injury to adjacent tissues when compared with standard stainless steel and titanium bipolar forceps. Of the two, heat-pipe embedded technology appeared safest, having better cooling efficiency at higher temperature.


The validity of the SF-12 and SF-6D instruments in people living with HIV/AIDS in Kenya.
Health-related quality of life (HRQoL) and health state utility value (HSUV) measurements are vital components of healthcare clinical and economic evaluations. Accurate measurement of HSUV and HRQoL require validated instruments. The 12-item Short-Form Health Survey (SF-12) is one of few instruments that can evaluate both HRQoL and HSUV, but its validity has not been assessed in people living with HIV/AIDS (PLWHA) in east Africa, where the burden of HIV is high. This cross-sectional study used baseline data from a randomized trial involving PLWHA in Kenya. Data included responses from a translated and adapted SF-12 survey as well as key demographic and clinical data. Construct validity of the survey was examined by testing the SF-12's ability to distinguish between groups known in advance to have differences in their health based on their disease severity. We classified disease severity based on established definitions from the US Center for Disease Control (CDC) and WHO, as well as a previously studied viral load threshold. T-tests and ANOVA were used to test for differences in HRQoL and HSUV scores. Area under the receive operator curve (AUC) was used to test the discriminative ability of the HRQoL and HSUV instruments. Differences in physical component scores met the minimum clinically important difference among participants with more advanced HIV when defined by CD4 count (4.3 units) and WHO criteria (compared to stage 1, stages 2, 3 and 4 were 2.0, 7.2 and 9.8 units lower respectively). Mental score differences met the minimum clinically important difference between WHO stage 1 and stage 4 patients (4.4). Differences in the HSUV were statistically lower in more advanced HIV by all three definitions of severity. The AUC showed poor to weak discriminatory ability in most analyses, but had fair discriminatory ability between WHO clinical stage 1 and clinical stage 4 individuals (AUC = 0.71). Our findings suggest that the Kiswahili translated and adapted version of the SF-12 could be used as an assessment tool for physical health, mental health and HSUV for Kiswahili-speaking PLHWA. Clinical trials.gov identifier: NCT00830622 . Registered 26 January 2009.


Finding a policy solution to India's diabetes epidemic.
In India, thirty-five million people have diabetes-a number expected to more than double by 2025, disproportionately affecting working-age people. The economic impact of this increase could be devastating to India's emerging economy. In this paper we discuss drivers of the epidemic, analyze current policies and practices in India, and conclude with recommendations, focusing on multisectoral and international collaboration. We see these recommendations as providing a blueprint for addressing diabetes in India by illuminating opportunities and barriers for policymakers and others.


The impact of histopathology and NAB2-STAT6 fusion subtype in classification and grading of meningeal solitary fibrous tumor/hemangiopericytoma.
Meningeal solitary fibrous tumor (SFT)/hemangiopericytoma (HPC) is a rare tumor with propensity for recurrence and metastasis. Although multiple classification schemes have been proposed, optimal risk stratification remains unclear, and the prognostic impact of fusion status is uncertain. We compared the 2016 WHO CNS tumor grading scheme (CNS-G), a three-tier system based on histopathologic phenotype and mitotic count, to the 2013 WHO soft-tissue counterpart (ST-G), a two-tier system based on mitotic count alone, in a cohort of 133 patients [59 female, 74 male; mean age 54 years (range 20-87)] with meningeal SFT/HPC. Tumors were pathologically confirmed through review of the first tumor resection (n = 97), local recurrence (n = 35), or distant metastasis (n = 1). A STAT6 immunostain showed nuclear expression in 132 cases. NAB2-STAT6 fusion was detected in 99 of 111 successfully tested tumors (89%) including the single STAT6 immunonegative tumor. Tumors were classified by CNS-G as grade 1 (n = 43), 2 (n = 41), or 3 (n = 49), and by ST-G as SFT (n = 84) or malignant SFT (n = 49). Necrosis was present in 16 cases (12%). On follow-up, 42 patients had at least one subsequent recurrence or metastasis (7 metastasis only, 33 recurrence only, 2 patients had both). Twenty-nine patients died. On univariate analysis, necrosis (p = 0.002), CNS-G (p = 0.01), and ST-G (p = 0.004) were associated with recurrence-free (RFS) but not overall survival (OS). NAB2-STAT6 fusion type was not significantly associated with RFS or OS, but was associated with phenotype. A modified ST-G incorporating necrosis showed higher correlation with RFS (p = 0.0006) and remained significant (p = 0.02) when considering only the primary tumors. From our data, mitotic rate and necrosis appear to stratify this family of tumors most accurately and could be incorporated in a future grading scheme.


Previously unrecognized stages of species-specific colonization in the mutualism between Xenorhabdus bacteria and Steinernema nematodes.
The specificity of a horizontally transmitted microbial symbiosis is often defined by molecular communication between host and microbe during initial engagement, which can occur in discrete stages. In the symbiosis between Steinernema nematodes and Xenorhabdus bacteria, previous investigations focused on bacterial colonization of the intestinal lumen (receptacle) of the nematode infective juvenile (IJ), as this was the only known persistent, intimate and species-specific contact between the two. Here we show that bacteria colonize the anterior intestinal cells of other nematode developmental stages in a species-specific manner. Also, we describe three processes that only occur in juveniles that are destined to become IJs. First, a few bacterial cells colonize the nematode pharyngeal-intestinal valve (PIV) anterior to the intestinal epithelium. Second, the nematode intestine constricts while bacteria initially remain in the PIV. Third, anterior intestinal constriction relaxes and colonizing bacteria occupy the receptacle. At each stage, colonization requires X. nematophila symbiosis region 1 (SR1) genes and is species-specific: X. szentirmaii, which naturally lacks SR1, does not colonize unless SR1 is ectopically expressed. These findings reveal new aspects of Xenorhabdus bacteria interactions with and transmission by theirSteinernema nematode hosts, and demonstrate that bacterial SR1 genes aid in colonizing nematode epithelial surfaces.


Comparative toxicity of valproic acid and its metabolites in liver slices from adult rats, weanling rats and humans.
The incidence of fatal hepatic failure associated with valproic acid (VPA) treatment is low but when it occurs, 90% of the affected patients are below the age of 20 yr. At present the mechanism of VPA hepatotoxicity is unclear; however, it may be a combination of the formation and action of toxic metabolites in developing tissues. In the study reported here we investigated the action of VPA and its metabolites in liver slices prepared from adult and weanling Sprague-Dawley rats and from human livers (non-transplantable livers from organ donors, or biopsy material from patients undergoing surgical liver resection). VPA, 2-propyl-2-pentenoic acid (2-en-VPA or Delta(2)-VPA), 2-propyl-4-pentenoic acid (4-en-VPA or Delta(4)-VPA) were incubated for various times at concentrations of 100 or 300 mug/ml. Protein synthesis and K(|) content were used to assess functional integrity or general viability. The question addressed was whether there were differences in the in vitro toxicity of VPA and its metabolites that were related to the age of the livers from which the slices were taken. Liver slices from weanling rats were significantly more sensitive to VPA and its metabolites than the slices from livers of adult rats. The rank order of toxicities (4-en-VPA > VPA > 2-en-VPA) was the same in both sets of rat slices. The human liver slices were significantly affected by VPA and its metabolites but these compounds were equal in their ability to produce this toxicity. There was also an indication of differences in sensitivity to the toxicity of VPA in slices from livers of donors of different ages.


The p160/steroid receptor coactivator family: potent arbiters of uterine physiology and dysfunction.
The p160/steroid receptor coactivator (SRC) family comprises three pleiotropic coregulators (SRC-1, SRC-2, and SRC-3; otherwise known as NCOA1, NCOA2, and NCOA3, respectively), which modulate a wide spectrum of physiological responses and clinicopathologies. Such pleiotropy is achieved through their inherent structural complexity, which allows this coregulator class to control both nuclear receptor and non-nuclear receptor signaling. As observed in other physiologic systems, members of the SRC family have recently been shown to play pivotal roles in uterine biology and pathobiology. In the murine uterus, SRC-1 is required to launch a full steroid hormone response, without which endometrial decidualization is markedly attenuated. From "dovetailing" clinical and mouse studies, an isoform of SRC-1 was recently identified which promotes endometriosis by reprogramming endometrial cells to evade apoptosis and to colonize as endometriotic lesions within the peritoneal cavity. The endometrium fails to decidualize without SRC-2, which accounts for the infertility phenotype exhibited by mice devoid of this coregulator. In related studies on human endometrial stromal cells, SRC-2 was shown to act as a molecular "pacemaker" of the glycolytic flux. This finding is significant because acceleration of the glycolytic flux provides the necessary bioenergy and biomolecules for endometrial stromal cells to switch from quiescence to a proliferative phenotype, a critical underpinning in the decidual progression program. Although studies on uterine SRC-3 function are in their early stages, clinical studies provide tantalizing support for the proposal that SRC-3 is causally linked to endometrial hyperplasia as well as with endometrial pathologies in patients diagnosed with polycystic ovary syndrome. This proposal is now driving the development and application of innovative technologies, particularly in the mouse, to further understand the functional role of this elusive uterine coregulator in normal and abnormal physiologic contexts. Because dysregulation of this coregulator triad potentially presents a triple threat for increased risk of subfecundity, infertility, or endometrial disease, a clearer understanding of the individual and combinatorial roles of these coregulators in uterine function is urgently required. This minireview summarizes our current understanding of uterine SRC function, with a particular emphasis on the next critical questions that need to be addressed to ensure significant expansion of our knowledge of this underexplored field of uterine biology.


A clonal expression biomarker associates with lung cancer mortality.
An aim of molecular biomarkers is to stratify patients with cancer into disease subtypes predictive of outcome, improving diagnostic precision beyond clinical descriptors such as tumor stage1. Transcriptomic intratumor heterogeneity (RNA-ITH) has been shown to confound existing expression-based biomarkers across multiple cancer types2-6. Here, we analyze multi-region whole-exome and RNA sequencing data for 156 tumor regions from 48 patients enrolled in the TRACERx study to explore and control for RNA-ITH in non-small cell lung cancer. We find that chromosomal instability is a major driver of RNA-ITH, and existing prognostic gene expression signatures are vulnerable to tumor sampling bias. To address this, we identify genes expressed homogeneously within individual tumors that encode expression modules of cancer cell proliferation and are often driven by DNA copy-number gains selected early in tumor evolution. Clonal transcriptomic biomarkers overcome tumor sampling bias, associate with survival independent of clinicopathological risk factors, and may provide a general strategy to refine biomarker design across cancer types.


Factors affecting the quality of life before and after surgery in patients with chronic thromboembolic pulmonary hypertension.
To test the hypothesis that quality of life (QoL) parameters before surgery in patients with chronic thromboembolic pulmonary hypertension (CTEPH) are influenced by clinical determinants related to the underlying disease and to examine QoL parameters affected in the long-term after the operation by complications presenting in the early postoperative period. This prospective cohort study included 128 patients who presented with CTEPH before and after pulmonary thromboendarterectomy (PTE; 1-year follow-up). All patients were examined regarding QoL using the Short-Form 36 Health Survey Questionnaire (SF-36). In patients with CTEPH, PTE provided immediate improvement in terms of pulmonary hemodynamic parameters and favorable effects on long-term outcome, including QoL 1 year after surgery. Multivariate analysis showed that systolic pulmonary arterial pressure, right ventricular ejection fraction, and the presence of coronary artery disease and chronic obstructive pulmonary disease (COPD) were independent factors affecting QoL on several SF-36 subscales in patients with CTEPH prior to surgery. The factors that affect patient QoL 1 year after surgery on some SF-36 subscales included the presence of coronary artery disease, COPD, heart failure, residual pulmonary hypertension, and prolonged ventilation, neurological complications in the early postoperative period of PTE. Surgical treatment for CTEPH leads to an increase in QoL in all SF-36 subscales, excluding general health perceptions. Factors affecting QoL in patients with CTEPH included severity of pulmonary arterial hypertension, comorbidity, and complications in the early postoperative period after PTE, such as heart failure, neurologic problems, residual pulmonary hypertension, and prolonged ventilation.


Precise design of copolymer-conjugated nanocatalysts for active electron transfer.
A copolymer-conjugated nanocatalytic system has been designed for active electron transfer. To enhance photoinduced H2 generation, we precisely synthesize ternary random copolymers capable of transferring electrons through phase transitions, extending and shrinking in response to viologen's redox changes within 2 nm distance from the surface of the catalytic nanoparticle.


Empirical investigation of friction weakening of terrestrial and Martian landslides using discrete element models.
Understanding what controls the travelling distance of large landslides has been the topic of considerable debate. By combining observation and experimental data with depth-averaged continuum modelling of landslides and generated seismic waves, it was empirically observed that lower effective friction had to be taken into account in the models to reproduce the dynamics and runout distance of larger volume landslides. Moreover, such simulation and observation results are compatible with a friction weakening with velocity as observed in earthquake mechanics. We investigate here as to whether similar empirical reduced friction should be put into discrete element models (DEM) to reproduce observed runout of large landslides on Earth and on Mars. First we show that, in the investigated parameter range and for a given volume, the runout distance simulated by 3D DEM is not much affected by the number (i.e. size) of grains once this number attains ~ 8000. We then calibrate the model on laboratory experiments and simulate other experiments of granular flows on inclined planes, making it possible for the first time to reproduce the observed effect of initial volume and aspect ratio on runout distances. In particular, the normalised runout distance starts to depend on the volume involved only above a critical slope angle > 16-19°, as observed experimentally. Finally, based on field data (volume, topography, deposit), we simulate a series of landslides on simplified inclined topography. The empirical friction coefficient, calibrated to reproduce the observed runout for each landslide, is shown to decrease with increasing landslide volume (or velocity), going down to values as low as 0.1-0.2. No distinguishable difference is observed between the behaviour of terrestrial and Martian landslides.


Cerebrospinal fluid myelin basic protein as a prognostic biomarker in dogs with thoracolumbar intervertebral disk herniation.
Release of myelin basic protein (MBP) into the cerebrospinal fluid (CSF) is associated with active demyelination and correlates with outcome in various neurological diseases. To describe associations among CSF MBP concentration, initial neurological dysfunction, and long-term ambulatory outcome in dogs with acute thoracolumbar intervertebral disk herniation (IVDH). Fifty seven [corrected] dogs with acute thoracolumbar IVDH and 16 clinically normal dogs. Prospective case series clinical study. Signalment, initial neurological dysfunction as determined by a modified Frankel score (MFS), and ambulatory outcome at >3-month follow-up were recorded. Cisternal CSF MBP concentration was determined by an ELISA. Associations were estimated between CSF MBP concentration and various clinical parameters. Dogs with thoracolumbar IVDH that did not ambulate at follow-up had a higher CSF MBP concentration (median, 3.56 ng/mL; range, 0.59-51.2 ng/mL) compared with control dogs (median, 2.22 ng/mL; range, 0-3.82 ng/mL) (P=.032). A CSF MBP concentration of >or=3 ng/mL had a sensitivity of 78% and specificity of 76% to predict an unsuccessful outcome based on receiver-operating characteristics curve analysis (area under the curve=0.688, P=.079). Affected dogs with a CSF MBP concentration>or=3 ng/mL had 0.09 times the odds of ambulation at follow-up compared with affected dogs with CSF MBP concentration<3 ng/mL when adjusted for initial MFS (95% confidence interval 0.01-0.66, P=.018). These results would suggest that CSF MBP concentration may be useful as an independent prognostic indicator in dogs with thoracolumbar IVDH.


The effect of cone beam CT (CBCT) on therapeutic decision-making in endodontics.
The aim was to assess to what extent cone beam CT (CBCT) used in accordance with current European Commission guidelines in a normal clinical setting has an impact on therapeutic decisions in a population referred for endodontic problems. The study includes data of consecutively examined patients collected from October 2011 to December 2012. From 2 different endodontic specialist clinics, 57 patients were referred for a CBCT examination using criteria in accordance with current European guidelines. The CBCT examinations were performed using similar equipment and standardized among clinics. After a thorough clinical examination, but before CBCT, the examiner made a preliminary therapy plan which was recorded. After the CBCT examination, the same examiner made a new therapy plan. Therapy plans both before and after the CBCT examination were plotted for 53 patients and 81 teeth. As four patients had incomplete protocols, they were not included in the final analysis. 4% of the patients referred to endodontic clinics during the study period were examined with CBCT. The most frequent reason for referral to CBCT examination was to differentiate pathology from normal anatomy, this was the case in 24 patients (45% of the cases). The primary outcome was therapy plan changes that could be attributed to CBCT examination. There were changes in 28 patients (53%). CBCT has a significant impact on therapeutic decision efficacy in endodontics when used in concordance with the current European Commission guidelines.


Involvement of the Fanconi's anemia protein FAC in a pathway that signals to the cyclin B/cdc2 kinase.
Lymphoblastoid cell lines derived from patients with the chromosomal instability disorder Fanconi's anemia (FA) are hyperresponsive to G2 delay and apoptosis induced by cross-linking agents such as mitomycin C (MMC). Here, we investigated whether the protein defective in FA complementation group C (FA-C) cells functions in a pathway that signals to the cdc2 kinase complex, which controls mitotic progression. FA-C lymphoblasts treated with a low dose of MMC (1-5 microM, 1 h) exhibited a protracted G2-M arrest and subsequent apoptosis by 2 days after treatment. This G2-M arrest was mediated by persistent inactivation of the cyclin B1/cdc2 kinase complex characterized by both sustained accumulation of cyclin B1 and tyrosine phosphorylation of cdc2. In phenotypically corrected (wild-type) cells, the same treatment induced only temporal G2-M arrest, associated with a transient inactivation of the cyclin B1/cdc2 kinase complex, after which cells resumed cycling. Treatment with higher dosages (15-30 microM, 1 h) resulted in S-phase arrest and induced a similar high level of apoptosis in FA-C and wild-type cells, accompanied by degradation of cyclin B1 and dephosphorylation of cdc2. In low-dose treated G2-M-arrested FA-C cells, caffeine-dependent activation of cdc2 released the G2-M block but failed to protect against apoptosis, suggesting that apoptosis was not a direct consequence of persistent cdc2 kinase inactivation. Thus, at low doses of MMC, FA-C cells exhibit a unique cyclin B1/cdc2 response that is not observed in wild-type cells treated with an equitoxic high dosage of cross-linker. Although these results do not necessarily implicate a role for FAC in regulating cyclin B/cdc2 kinase activity, available evidence suggests that the FAC protein is involved in a cross-link damage avoidance pathway that signals to this kinase complex.


Prevalence of obesity and metabolic syndrome in Indigenous Australian youths.
We conducted a cross-sectional study of Indigenous youths residing in the Torres Strait region of Australia to assess the prevalence of obesity and the metabolic syndrome. Data on body mass index (BMI), waist circumference, blood pressure, presence of acanthosis nigricans and blood glucose were collected. Fasting glucose, insulin, C-Peptide, HbA1c and lipids were measured, and an oral glucose tolerance test was performed in those with a BMI greater than 25 (childhood-equivalent cut-points) or fasting glucometer reading >5.5 mmol/L. Of 158 youths, 31% were overweight and 15% were obese, 38% had enlarged waist circumference consistent with central obesity, 43% had acanthosis nigricans and 27% were hypertensive. More females than males had enlarged waist circumferences (59% vs. 13%, P < 0.001). Among overweight or obese youth, 56% had significantly elevated insulin (P = 0.021); they also had higher HOMA-IR (P = 0.002). The metabolic syndrome was present in 17% of all youths (mostly females) and in 33% of the overweight or obese subgroup. Type 2 diabetes was diagnosed in two youths. These very high proportions of overweight or obese Torres Strait youth with metabolic risk factors have major public health implications.


Khat Chewing and Type 2 Diabetes Mellitus.
Knowledge about the effects of khat chewing on type 2 diabetes mellitus (T2DM) development and glycemic control is very sparse. Emerging data suggest that khat chewing may increase the risk of T2DM occurrence. Therefore, this study aimed to measure the prevalence of khat chewing in Saudi people with T2DM in Jazan, Saudi Arabia and to determine the association of khat chewing with T2DM development and glycemic control in T2DM. This is an analytical, cross-sectional study that included 472 Saudi participants selected randomly from primary healthcare centers in Jazan, Saudi Arabia. A chi-square test and logistic regression were performed in the statistical analysis. The prevalence of khat chewing in Saudi patients with T2DM in Jazan was 29.3%. After adjusting for covariates, khat chewing was significantly associated with T2DM (odds ratio 3.5), indicating that khat chewers had a more than three times higher risk of developing T2DM than those who do not chew khat. However, there was no association between khat chewing and glycemic control in T2DM. Khat chewing was highly prevalent in Saudi people with T2DM in Jazan, Saudi Arabia. There was an association between khat chewing and the development of T2DM. Establishing the causal association of khat chewing with T2DM development and glycemic control and clarifying the biological role of khat in T2DM are important aims for future studies.


Behavioral changes following a single episode of early-life seizures support the latent development of an autistic phenotype.
We probed the developmental and behavioral consequences of a single episode of kainic acid-induced early-life seizures (KA-ELS) in the rat on postnatal day 7. Correlates of developmental trajectory were not altered, demonstrating that long-term consequences following KA-ELS are not initiated by secondary causes, such as malnourishment or alterations in maternal care. We report reduced marble burying in adult rats, suggestive of restricted interests, a trait common to experimental and clinical autism. We did not detect increased repetitive grooming during habituated cage behavior. However, we did detect reduced grooming in adult KA-ELS rats in the presence of an unfamiliar rat, supporting altered social anxiety following KA-ELS. Reanalysis of a social approach task further indicated abnormal social interactions. Taken together with previous physiological and behavioral data, these data support the hypothesis that KA-ELS lead to a latent autistic phenotype in adult rats not attributable to other early alterations in development.


On-call hospital pharmacy services: a perspective from NHS Tayside, Scotland.
To describe the enquiries to an out-of-hours pharmacy on-call service and to describe the activity of this service from April 2015 to March 2016. Data entered by on-call pharmacists into the 'On Call' app database at a large teaching hospital were reviewed retrospectively. Data were analysed using OpenOffice Calc. 839 on-call enquiries were received in the data collection period, averaging 70 calls per month or 2.3 calls per shift. The busiest days were Saturdays (26.0% of total calls, 218/839) and Sundays (20.0% of total calls, 168/839). The vast majority of calls were received before midnight (91.8%, 770/839), with most of these being received between 09.00 and 20.00 (68.1%, 571/839). The number of calls varied greatly between months with the least calls being received in April (37) and the most in January (100). Nurses were the main users of the service with 62.8% (527/839) calls. Junior doctors were the other main users with 26.2% (220/839) of calls. Most calls involved queries regarding the supply of medicines (30.0%, 252/839), Medicines information requests (26.8%, 225/839) or requests to dispense discharge prescriptions (17.6%, 148/839). The majority of calls were resolved within 30 min (82.4%, 691/839) and a significant proportion answered within 10 min (48.6%, 408/839). The average time taken to resolve a call was 22 min. More experienced members of the service were able to resolve calls more quickly than the more junior (averages of 26.64 vs 19.73 min). This article provides a commentary on the pharmacy on-call service within NHS Tayside and an in-depth look at what an on-call service entails.