CD98hc (SLC3A2) loss protects against ras-driven tumorigenesis by modulating integrin-mediated mechanotransduction.
CD98hc (SLC3A2) is the heavy chain component of the dimeric transmembrane glycoprotein CD98, which comprises the large neutral amino acid transporter LAT1 (SLC7A5) in cells. Overexpression of CD98hc occurs widely in cancer cells and is associated with poor prognosis clinically, but its exact contributions to tumorigenesis are uncertain. In this study, we showed that genetic deficiency of CD98hc protects against Ras-driven skin carcinogenesis. Deleting CD98hc after tumor induction was also sufficient to cause regression of existing tumors. Investigations into the basis for these effects defined two new functions of CD98hc that contribute to epithelial cancer beyond an intrinsic effect of CD98hc on tumor cell proliferation. First, CD98hc increased the stiffness of the tumor microenvironment. Second, CD98hc amplified the capacity of cells to respond to matrix rigidity, an essential factor in tumor development. Mechanistically, CD98hc mediated this stiffness sensing by increasing Rho kinase (ROCK) activity, resulting in increased transcription mediated by YAP/TAZ, a nuclear relay for mechanical signals. Our results suggest that CD98hc contributes to carcinogenesis by amplifying a positive feedback loop, which increases both extracellular matrix stiffness and resulting cellular responses. This work supports a rationale to explore the use of CD98hc inhibitors as cancer therapeutics.


Minisatellite polymorphism as a tool to distinguish closely related Lactococcus lactis strains.
Genome plasticity is considered as a means for bacteria to adapt to their environment. Plasticity in tandem repeat sequences on bacterial genomes has been recently exploited to trace the epidemiology of pathogens. Here, we examine the utility of minisatellite (i.e., a repeat unit of six nucleotides or more) typing in non-pathogenic food bacteria of the species Lactococcus lactis. Thirty-four minisatellites identified on the sequenced L. lactis ssp. lactis strain IL1403 genome were first analyzed in 10 closely related ssp. lactis strains, as determined by randomly amplified polymorphic DNA (RAPD). The selected tandem repeats varied in length, percent identity between repeats, and locations. We showed that: (i) the greatest polymorphism was in orfs encoding exported proteins or in intergenic regions; (ii) two thirds of minisatellites were little- or non-variable, despite as much as 90% identity between tandem repeats; and (iii) dendrograms based on either RAPD or minisatellite analyses were similar. Seven minisatellites identified in this study are potentially useful for lactococcal typing. We then asked whether tandem repeats in L. lactis were stable upon very long-term (up to two years) storage. Despite large rearrangements previously reported in derivative strains, just one of 10 minisatellites tested underwent an alteration, suggesting that tandem repeat rearrangements probably occur during active DNA replication. We conclude that multiple locus minisatellite analysis can be a valuable tool to follow lactococcal strain diversity.


Evaluation of Irradiated Corneas Using Scatterometry and Light and Electron Microscopy.
To determine if sterilization of donor corneas by irradiation alters optical, histologic, or ultrastructural tissue characteristics. Blinded assessment of 10 irradiated and 10 nonirradiated donor corneas by a scatterometer used to objectively measure backscattered corneal light. Light and electron microscopy were performed on samples from both groups. The mean scattering of the nonirradiated (control) corneas was significantly lower (0.0060 ± 0.0034, mean ± SD) than the scattering of the irradiated corneas (0.023 ± 0.0078; P = 0.0001). There was no statistical difference between ages, days in storage, or central corneal thickness of the 2 groups. Light microscopic evaluation revealed attenuation of the irradiated corneal epithelium with superficial stromal clefting. Transmission electron microscopy demonstrated similar fibril diameter in the nonirradiated and irradiated corneas. Although the mean interfibrillar distance was similar between the nonirradiated and irradiated groups, the interfibrillar distance in the superficial stroma was increased in the nonirradiated compared with the irradiated groups (15.4 ± 2.4 vs. 11.8 ± 0.98 nm; P ≤ 0.05). Our data quantitatively support increased light scatter after irradiation, which cannot be attributed to increased corneal thickness. Irradiation alters the histology of the epithelium and the ultrastructure of the superficial corneal stroma. Increased light scatter because of epithelial changes would not be expected to impact postoperative visual function. However, increased light scatter because of stromal changes may affect postoperative visual function if used for optical corneal rehabilitation. Further investigation is warranted to determine whether attenuated epithelium or superficial stromal changes are the cause of increased light scatter in the irradiated corneal tissue.


Identification of adenoviruses in specimens from high-risk pediatric stem cell transplant recipients and controls.
Adenovirus infection is an important cause of morbidity and mortality in stem cell transplant recipients. We report species and type-specific analysis from a prospective study of high-risk adenovirus infections following hematopoietic progenitor cell transplantation prior to, during, and after treatment with cidofovir, as well as species analysis of contemporaneously collected samples from control patients. Nine different adenovirus types representing all six recognized species were identified, and mixed infections were commonly found in this group of patients.


How to measure responses of the knee to lateral perturbations during gait? A proof-of-principle for quantification of knee instability.
Knee instability is a major problem in patients with anterior cruciate ligament injury or knee osteoarthritis. A valid and clinically meaningful measure for functional knee instability is lacking. The concept of the gait sensitivity norm, the normalized perturbation response of a walking system to external perturbations, could be a sensible way to quantify knee instability. The aim of this study is to explore the feasibility of this concept for measurement of knee responses, using controlled external perturbations during walking in healthy subjects. Nine young healthy participants walked on a treadmill, while three dimensional kinematics were measured. Sudden lateral translations of the treadmill were applied at five different intensities during stance. Right knee kinematic responses and spatio-temporal parameters were tracked for the perturbed stride and following four cycles, to calculate perturbation response and gait sensitivity norm values (i.e. response/perturbation) in various ways. The perturbation response values in terms of knee flexion and abduction increased with perturbation intensity and decreased with an increased number of steps after perturbation. For flexion and ab/adduction during midswing, the gait sensitivity norm values were shown to be constant over perturbation intensities, demonstrating the potential of the gait sensitivity norm as a robust measure of knee responses to perturbations. These results show the feasibility of using the gait sensitivity norm concept for certain gait indicators based on kinematics of the knee, as a measure of responses during perturbed gait. The current findings in healthy subjects could serve as reference-data to quantify pathological knee instability.


[Isolation and study of the properties of the regulator subunit of cAMP-dependent protein kinase].
The regulatory subunit of type II cAMP-dependent proteinkinase was isolated from cytosol of the rabbit small intestinal mucosa by affinity chromatography. The preparation contained 3 proteolytic enzymes and occurred in two forms differing as regards cAMP affinity. The cAMP-binding capacity of the preparation was equal to 17 nmol cAMP/mg protein. To study the topography of the cAMP-binding center, use was made of cAMP analogs. It was demonstrated that introduction of the substituents into the 8th position of the purine ring and substitution with respect to the N6-exoaminogroup affected insignificantly the analog affinity for the cAMP-binding center. At the same time the substituents introduced into the first position of the adenine base, into the area of the 2'-hydroxyl group of ribose and into the cyclophosphate part of the cAMP molecule considerably decreased the analog affinity for the regulatory center of type II cAMP-dependent proteinkinase.


The effect of cannabis in the treatment of Hodgkin's lymphoma in a pregnant patient - extensive case report and literature review.
Hodgkin lymphoma (HL) is the fourth most frequent cancer diagnosis among pregnant females. A multidisciplinary team is mandatory to obtain the best treatment and prognosis for the mother and for the baby. Here, we present the case of a patient diagnosed with HL and its evolution during 2 pregnancies. Herein we present the case of a 21-year-old female Caucasian patient, with free history, diagnosed with HL stage IIB. The patient started first line chemotherapy and radiotherapy, with incomplete remission. She refused any other treatment. Five years later, the patient became pregnant and was offered chemotherapy in the 2nd trimester of pregnancy, that she refused, and delivered by C-section at 37 weeks. In the same year, the patient became pregnant again and was proposed termination of pregnancy, that she also refused. The MRI scan revealed progression of HL and she was admitted in the hospital several times for altered general condition, respiratory infections and increased need of painkillers including opioids. At 26 weeks of pregnancy, the patient began on her own a treatment with pure cannabis. Her pain and general status got better and the tumor tissue decreased. She delivered by C-section at 34 weeks a boy that presented in the first 24 h postpartum a withdrawal syndrome and intestinal invagination, requiring care in NICU and surgery with bowel resection. Therefore, we can conclude that cannabis could be part of oncological treatment. No other case like this, as far as we know, has been previously reported.


Real World Experience of Drug Induced Liver Injury in Patients Undergoing Chemotherapy.
To better understand the clinical significance of drug induced liver injury (DILI) during chemotherapy, we examined the epidemiology, incidence, and treatment effects of DILI in patients undergoing chemotherapy for genitourinary malignancies over a two-year period. We conducted a retrospective review of 284 patients who underwent chemotherapy for prostate, bladder, testicular and renal cell carcinomas over a two year period. Those with abnormal or absent liver test (LT) results prior to chemotherapy initiation were excluded. Post chemotherapy LT results were defined as DILI if ALT>3× ULN and/or total bilirubin (TB)>2× ULN, in the absence of other more likely causes of elevated LT. The cumulative incidence of DILI in the total study population was 6.1% (17/284), and in the population who had appropriate LT performed it increased to 18.9% (17/90). Chemotherapeutic agents were determined to be the cause of DILI in 82% (14/17) of patients, and the treatment plans were changed in 59% (10/17) of patients. In this real world study, the cumulative incidence of DILI was higher than commonly reported in clinical trials, and the majority of affected patients had to have their cancer treatment altered or interrupted.


European Bluetongue Serotype 8: Disease Threat Assessment for U.S. Sheep.
Bluetongue virus (BTV) is an orbivirus transmitted by biting midges (Culicoides spp.) that can result in moderate to high morbidity and mortality primarily in sheep and white-tailed deer. Although only 5 serotypes of BTV are considered endemic to the United States, as many as 11 incursive serotypes have been detected in livestock and wildlife in the past 16 years. Introductions of serotypes, with unknown virulence and disease risk, are constant threats to US agriculture. One potential incursive serotype of particular concern is the European strain of BTV-8, which was introduced into Northern Europe in 2006 and caused unprecedented livestock disease and mortality during the 2006-2007 vector seasons. To assess disease risk of BTV-8 in a common white-faced American sheep breed, eight Polled Dorset yearlings were experimentally infected and monitored for clinical signs. Viremia and viral tissue distribution were detected and quantified by real-time qRT-PCR. Overall, clinical disease was moderate with no mortality. Viremia reached as high as 9.7 log10 particles/mL and persisted at 5 logs or higher through the end of the study (28 days). Virus distribution in tissues was extensive with the highest mean titers at the peak of viremia (day 8) in the kidney (8.38 log10 particles/mg) and pancreas (8.37 log10 particles/mg). Virus persisted in tissues of some sheep at 8 logs or higher by day 28. Results of this study suggest that should BTV-8 emerge in the United States, clinical disease in this common sheep breed would likely be similar in form, duration, and severity to what is typically observed in severe outbreaks of endemic serotypes, not the extraordinary disease levels seen in Northern Europe. In addition, a majority of exposed sheep would be expected to survive and act as significant BTV-8 reservoirs with high titer viremias for subsequent transmission to other livestock and wildlife populations.


[Disorders of consciousness: the basis for ethical assessment].
During the past few years there is an ethical debate about neurological disease entities that are characterised by a) prolongation of life owing to medical treatment, b) limited chances of cure, and c) impaired to unbearable life quality: akinetic mutism, vegetative state ("Wachkoma", apallic syndrome), and locked-in syndrome. These are compared to typical coma and brain death. According to Gerstenbrand (1967) [34] the vegetative state is differentiated into the transitional state following typical coma, the variations of typical and incomplete vegetative state, the remission state and the "Durchgangssyndrom" (characterized by preserved wakefulness with affective lability, disorientation, and amnesia). With regard to pathogenesis we differentiate posttraumatic and posthypoxic origin and variable lesions in cerebral cortex, thalamus or mesencephalic reticular formation. Uncertainty of prognosis is stressed. In respect of brain death we compare a) neocortical death, b) brain stem death, and c) whole brain death, and discuss problems of difficult delimitation and uncertainty of diagnosis. These syndromes are compared to anencephaly and hydranencephaly. Regarding the locked-in syndrome, typical, incomplete and complete (total) forms are distinguished. The differential diagnosis between the complete locked-in syndrome and brain stem death seems problematic. Difficulties in decisions limiting therapeutic effort stem from a) essentially intuitive judgement about observations, b) variability of syndromes and courses, c) uncertainty of prognosis, and d) differences in understanding and valuation throughout society. Emphasis is on a trustful and open colloquy among the persons concerned.


A study of nurses who completed a hospital-based reentry program.
This article describes a survey of nurses who completed a reentry program. The study resulted in information about demographics, job history, career development, and career satisfaction. Findings reveal that the reentry program was successful in assisting inactive nurses to return to the work force.


Practical Analysis of Genome Contact Interaction Experiments.
The three dimensional (3D) architecture of chromosomes is not random but instead tightly organized due to chromatin folding and chromatin interactions between genomically distant loci. By bringing genomically distant functional elements such as enhancers and promoters into close proximity, these interactions play a key role in regulating gene expression. Some of these interactions are dynamic, that is, they differ between cell types, conditions and can be induced by specific stimuli or differentiation events. Other interactions are more structural and stable, that is they are constitutionally present across several cell types. Genome contact interactions can occur via recruitment and physical interaction between chromatin-binding proteins and correlate with epigenetic marks such as histone modifications. Absence of a contact can occur due to presence of insulators, that is, chromatin-bound complexes that physically separate genomic loci. Understanding which contacts occur or do not occur in a given cell type is important since it can help explain how genes are regulated and which functional elements are involved in such regulation. The analysis of genome contact interactions has been greatly facilitated by the relatively recent development of chromosome conformation capture (3C). In an even more recent development, 3C was combined with next generation sequencing and led to Hi-C, a technique that in theory queries all possible pairwise interactions both within the same chromosome (intra) and between chromosomes (inter). Hi-C has now been used to study genome contact interactions in several human and mouse cell types as well as in animal models such as Drosophila and yeast. While it is fair to say that Hi-C has revolutionized the study of chromatin interactions, the computational analysis of Hi-C data is extremely challenging due to the presence of biases, artifacts, random polymer ligation and the huge number of potential pairwise interactions. In this chapter, we outline a strategy for analysis of genome contact experiments based on Hi-C using R and Bioconductor.


Low-volume plus ascorbic acid vs high-volume plus simethicone bowel preparation before colonoscopy.
To investigate the effectiveness of low-volume plus ascorbic acid [polyethylene glycol plus ascorbic acid (PEG + Asc)] and high-volume plus simethicone [polyethylene glycol plus simethicone (PEG + Sim)] bowel preparations. A total of one hundred and forty-four outpatients (76 males), aged from 20 to 84 years (median age 59.5 years), who attended our Department, were divided into two groups, age and sex matched, and underwent colonoscopy. Two questionnaires, one for patients reporting acceptability and the other for endoscopists evaluating bowel cleansing effectiveness according to validated scales, were completed. Indications, timing of examination and endoscopical findings were recorded. Biopsy forceps were used as a measuring tool in order to determine polyp endoscopic size estimation. Difficulty in completing the preparation was rated in a 5-point Likert scale (1 = easy to 5 = unable). Adverse experiences (fullness, cramps, nausea, vomiting, abdominal pain, headache and insomnia), number of evacuations and types of activities performed during preparation (walking or resting in bed) were also investigated. Seventy-two patients were selected for each group. The two groups were age and sex matched as well as being comparable in terms of medical history and drug therapies taken. Fourteen patients dropped out from the trial because they did not complete the preparation procedure. Ratings of global bowel cleansing examinations were considered to be adequate in 91% of PEG + Asc and 88% of PEG + Sim patients. Residual Stool Score indicated similar levels of amount and consistency of residual stool; there was a significant difference in the percentage of bowel wall visualization in favour of PEG + Sim patients. In the PEG + Sim group, 12 adenomas ≤ 10 mm diameter (5/left colon + 7/right colon) vs 9 (8/left colon + 1/right colon) in the PEG + Asc group were diagnosed. Visualization of small lesions seems to be one of the primary advantages of the PEG + Sim preparation. PEG + Asc is a good alternative solution as a bowel preparation but more improvements are necessary in order to achieve the target of a perfect preparation.


Peripheral facial palsy: muscle activity in different onset times.
To assess, through surface electromyography (sEMG), the activity of the risorius and zygomaticus muscles, during the production of voluntary smiles and to compare these data between two groups of individuals with different onset times of peripheral facial palsy (PFP). A total of 140 adults were divided into three groups: G1 (35 individuals with PFP onset time between 0 and 3 months); G2 (35 individuals with PFP onset time between 3 and 6 months); CG (control group) (70 healthy controls). All of the participants were submitted to the following assessments: clinical protocol for the assessment of facial mimic and sEMG of the risorius and zygomaticus muscles. The results suggest that the groups of individuals with PFP differed from the control group considering muscle activity during rest and during the production of voluntary smiles, regardless of the onset time of the disease. The groups with PFP did not differ between themselves in any of the tested situations. The group with PFP with longer onset time presented greater muscle activation asymmetry during the production of the voluntary smiles when compared to the other two groups. Muscle asymmetry was more evident when considering the results for the risorius muscle. The results of the sEMG do not distinguish the groups when considering PFP onset times.


Intratester and intertester reliability of goniometric measurement of passive lateral shoulder rotation.
Measurement of lateral rotation range of motion (ROM) is frequently performed during shoulder evaluation. The purpose of this study was to determine the intratester and intertester reliability of goniometric measurement of passive lateral rotation ROM of the shoulder. Two experienced PTs performed the testing in a randomized block design. They were blinded to all clinical information and to their goniometric readings. Passive lateral rotation ROM of the shoulder was assessed in 34 patients with a variety of shoulder pathologies. Patients were placed in the supine position with the arm abducted approximately 20 degrees to 30 degrees. A standard goniometer, placed along the joint axis by the therapist, was red by an independent assistant. Intraclass correlation coefficients (ICCs) and their associated 95% confidence intervals were calculated. Intratherapist ICCs (0.88 and 0.93) and intertherapist ICCs (0.85 and 0.80) were high. These findings suggest that reliable measures of passive lateral rotation ROM of the shoulder can be obtained from patients with shoulder pathology using standard goniometry and by placing the patient in a supine position.


A cross-sectional study on assessment of spiritual health and its associated factors among undergraduate students in a medical college in Mangalore, Karnataka.
Background: Spirituality and spiritual health are an integral component of an individual's health and wellbeing. Among medical undergraduates and professionals, spiritual health has positive effects on the individual by decreasing burnout, psychological distress, and rates of substance abuse. Spiritual health is also correlated with increased satisfaction and meaning in life. Spiritual health also influences future patient care, builds patient- provider communication channels, and improves patient outcomes. Methods: A cross-sectional study was conducted to assess the spiritual health levels of medical undergraduates in a medical college, in Karnataka, India, and to identify the factors associated with it. Using a pre-designed, validated semi-structed questionnaire, 600 medical undergraduates were approached and provided the forms. Spiritual health was assessed across the three domains of self-development, self-actualization, and self-realization using the Spiritual Health Assessment Scale (SHAS). Results: A total of 436 medical undergraduates participated. Majority (67.7%) of participants were girls. Most (62%) were found to have fair spiritual health with a large portion of the rest (36%) having good spiritual health and 2% had poor spiritual health. A significant association was found between spiritual health and the father's (p=0.005) and mother's (p=0.012) education levels. Spiritual health was also found to be associated with living in a nuclear family (p=0.04). Conclusions: Majority of the medical undergraduates had fair spiritual health. Parents' education levels and nuclear family were significantly associated with spiritual health.


Effects of luminal deoxynivalenol and L-proline on electrophysiological parameters in the jejunums of laying hens.
Most amino acids are cotransported with sodium. Deoxynivalenol (DON) decreases glucose absorption in the chicken small intestine in vivo and in vitro, and this effect is apparently mediated by the inhibition of the sodium D-glucose cotransporter. DON could selectively modulate the activities of other intestinal transporters. In order to assess this hypothesis, a study was conducted to characterize the in vitro effects of DON in the presence of mucosal amino acids, using L-proline as a model, on the electrophysiological parameters in the jejunums of laying hens. L-Proline (mucosal concentration of 1 mmol/L) was added to a stripped proximal part of jejunum sheets mounted in Ussing chambers in Ringer buffer, and the electrical properties were measured. The transmural potential difference (PD) was nearly constant between the treatments. The tissue resistance (Rt) was higher (P < 0.05) in the tissues exposed to DON compared with basal values and the values after addition of L-proline. Addition of L-proline on the luminal side of the isolated mucosa increased (P < 0.05) the short circuit-current (Isc), and it decreased (P < 0.05) after addition of DON, indicating that the proline-induced Isc was altered by DON. The addition of proline after incubation of the tissues with DON had no effect (P > 0.05) on PD or Rt. Proline did not increase the Isc under these conditions. DON decreased (P < 0.1) the Isc after addition of proline, indicating that DON inhibited the Na+-amino acid co-transport. We concluded from the present study that the amino acid cotransporter activity appears to be highly sensitive to DON suppression.


Soluble C-erbb-2 (p185) in breast-cancer patients in relation to prognosis.
Serum levels of P185-HER-2 were measured in 137 breast cancer patients and in 40 controls. The patients were divided into 4 groups: group A - 40 newly diagnosed patients; group B - 57 patients on long-term follow-up without active disease; group C - 26 patients with metastatic disease and group D - 5 patients with locally advanced inoperable tumors. The median level in controls was 4.8 U/l. The median P185 serum levels in groups C and D were significantly higher compared to groups A and B. In group C 60% and in group D 100% of patients had baseline elevated levels of serum P185 (>5 U/l) compared to 28% in groups A and B. Of the 14 patients in group A with elevated baseline levels of serum P185, 6 (43%) developed metastasis during the 24-month follow-up period. On serial measurements during follow-up in 23 patients of group A, 3 relapsed and the P185 level increased. In group C, serial measurements in patients with elevated baseline levels of P185 correlated with clinical response to therapy. These data suggest that serum levels of P185 are elevated in patients with metastatic disease. High initial P185 serum levels in new patients may have prognostic significance. Serial measurements of P185 in asymptomatic patients may help in monitoring disease state. In metastatic patients, serial P185 determination may be of benefit in assessing response to therapy.


Frequency of gonadal tumours in complete androgen insensitivity syndrome (CAIS): A retrospective case-series analysis.
Complete androgen insensitivity syndrome (CAIS) is an X-linked recessive disorder of sex development (DSD) where affected individuals are phenotypically female, but have an XY karyotype and testes. The risk of gonadal tumour development in CAIS may increase with age; incidence rates have been reported to be 0.8-22% in patients who have retained their gonads into adulthood. Consequently, gonadectomy has been recommended either during childhood or after puberty is complete, although there is no consensus on the optimal timing for this procedure. To establish the frequency of histological abnormalities in CAIS in relation to the age at gonadectomy. Data were collected from the Cambridge DSD database on patients with CAIS (n = 225; age range 3-88 years) who had undergone gonadectomy, and their age of gonadectomy, gonadal histology and immunohistochemistry. Evaluable data were obtained from 133 patients. Median age at gonadectomy was 14.0 years (range: 18 days-68 years). Pubertal status was: prepuberty, n = 62; postpuberty, n = 68. Thirteen cases were aged >20 years at gonadectomy. The pattern of histology is summarised in the Summary table. In this large case series of CAIS patients who had undergone gonadectomy, while the combined malignant and premalignant gonadal histology prevalence was 6.0%, the findings confirm the low occurrence of gonadal malignancy in CAIS, with a frequency of 1.5%. The two cases of malignancy were postpubertal. Germ cell neoplasia in situ (GCNIS) was observed in six cases, of which one occurred prepuberty and five postpuberty. The study highlighted difficulties in diagnosis of GCNIS and the need for histological analysis in expert centres. The results support the current recommendation that gonads in CAIS can be retained until early adulthood. The small number of individuals with gonadectomy after age 20 years do not allow firm conclusion regarding later adulthood. Therefore, it is recommended that the option of gonadectomy be discussed in adulthood. Some form of regular surveillance of the gonads is then recommended, although none of the available options are ideal.


Giant cell tumors of the spine: has denosumab changed the treatment paradigm?
OBJECT Giant cell tumors (GCTs) of the spine are rare and complex to treat. They have a propensity for local recurrence and the potential to metastasize. Treatment is currently surgical and presents unique challenges due to the proximity of neural structures and the need for reconstruction. Denosumab has been shown in clinical trials to be an effective treatment for GCT, but has not yet been studied specifically in GCT of the spine or as a surgical adjunct. To the authors' knowledge this is the first such reported series. METHODS A multicenter, prospective series of 5 patients with GCT of the spine treated with denosumab were included. Patient demographic data, oncological history, neurological status, tumor staging, treatment details and adverse events, surgical procedure, complications, radiological and histological responses, and patient outcome were analyzed. RESULTS All patients were women, with a mean age of 38 years, and presented with pain; 2 patients had additional neurological signs and symptoms. The mean duration of symptoms was 62 weeks. No patient had a prior tumor or metastatic disease at presentation. All patients had Enneking Stage III tumors and were treated with monthly cycles of 120 mg of denosumab, with initial additional loading doses on Days 8 and 15. Patients were given daily supplements of calcium (500 mg) and vitamin D (400 IU). There were no denosumab-related adverse events. All patients had a radiological response to denosumab. One patient failed to have a histological response to denosumab, with > 90% of tumor cells found to be viable on histological investigation. CONCLUSIONS This study reports the early experience of using denosumab in the treatment of spinal GCT. The results demonstrate a clinically beneficial radiological response and an impressive histological response in most but not all patients. Further experience with denosumab and longer patient follow-up is required. Denosumab has the potential to change the treatment paradigm for spinal GCT.


Anatomic resection for severe blunt liver trauma in 100 patients: significant differences between young and elderly.
The liver is the organ most commonly injured during blunt abdominal trauma. As our society ages, emergency surgery for active elderly patients increases, but data on aggressive emergency hepatic resection remain scarce in the literature. The purpose of this study was to determine whether the elderly (70 years of age or older) can tolerate major liver injury and subsequent hepatic resection. We investigated 100 patients who were treated by an anatomic resection for severe blunt liver trauma (29 elderly patients who were 70 years of age or older and 71 young patients who were younger than 70 years of age) in a retrospective study. The elderly patients were more severely injured as demonstrated by a higher Injury Severity Score, a lower Glascow Coma Scale, and lower survival (80.3% vs. 65.5%; p < 0.05). The total number of associated injuries was greater in elderly patients. Motor vehicle accidents were responsible for 71.8% of the injuries in the young group, and the predominant mechanism in the elderly patients was also motor vehicle accidents (51.7%). The 71 anatomic hepatic resections performed on the young patients included right hemihepatectomy (n = 45), left lateral segment resection (n = 14), bisegmentectomy (n = 5), and others. The 29 anatomic hepatic resections performed for the elderly patients were right hemihepatectomy (n = 15), left lateral segment resection (n = 5), left hemihepatectomy (n = 4), and others. Pneumonia, subphrenic abscess, and urosepsis occurred at a significantly higher frequency in elderly patients than in young patients. Our data clearly indicated that (1) the mechanism of injury, grade of associated intraabdominal injuries, distribution of surgical procedures, and complications differ significantly between young and elderly patients; and (2) the survival rate (65.5%) in elderly patients may be sufficient to consider anatomic hepatic resection to be a useful, safe procedure.


[EMOTIONAL DISORDERS IN CHILDREN VICTIMS OF NATURAL DISASTERS].
The effects of disasters on physical health tend to be well-known, with short, medium and long term sequelae. On the other hand, not always is have recognized in the same way the effects on mental health, despite having shown that, in situations of disaster or catastrophe there is a psychological signs of suffering increase and increases to a certain extent the psychiatric morbidity and other problem social. It is estimated that between a third and half of the exposed population, it suffers from some psychological manifestation. It has been erroneously thought that children and adolescents, not suffering with the same intensity of especially traumatic situations. In fact it was presumed, given their reactions so different from that of adults, had some protection. Currently, this has denied and minors are considered to be a group of high risk in cases of disasters and emergencies. Investigations carried out, demonstrate that in children and adolescents, the psychological sequels tend to be frequent and affect directly to the physical, mental and social development. Natural disasters are unexpected situations that will produce a serie of emotional reactions of diverse severity in their survivors, especially children, one of the most vulnerable groups due to a less understanding of what happened and difficulty expressing what they feel, having a personality still developing, and so directly affecting their physical, mental and social development. Therefore suffering the emotional scars, they will take longer to resolve and have a lifetime to live with them. These consequences should be treated by a corresponding community nurse and sometimes, depending on the severity and persistence (more than 3 months), a referral will be made to a qualified mental health professional, taking into account a number of recommendation and assesment canons. Parents or tutors with health professionals have an important role in the recovery of their children and their reactions will be very influenced by these firts. In recent years the interest has increased by the impact of the disaster on the mental health of affected populations and currently has become evident the need for a comprehensive approach that transcends disease care and repair of material damage.


The T box transcription factor TBX2 promotes epithelial-mesenchymal transition and invasion of normal and malignant breast epithelial cells.
The T box transcription factor TBX2, a master regulator of organogenesis, is aberrantly amplified in aggressive human epithelial cancers. While it has been shown that overexpression of TBX2 can bypass senescence, a failsafe mechanism against cancer, its potential role in tumor invasion has remained obscure. Here we demonstrate that TBX2 is a strong cell-autonomous inducer of the epithelial-mesenchymal transition (EMT), a latent morphogenetic program that is key to tumor progression from noninvasive to invasive malignant states. Ectopic expression of TBX2 in normal HC11 and MCF10A mammary epithelial cells was sufficient to induce morphological, molecular, and behavioral changes characteristic of EMT. These changes included loss of epithelial adhesion and polarity gene (E-cadherin, ß-catenin, ZO1) expression, and abnormal gain of mesenchymal markers (N-cadherin, Vimentin), as well as increased cell motility and invasion. Conversely, abrogation of endogenous TBX2 overexpression in the malignant human breast carcinoma cell lines MDA-MB-435 and MDA-MB-157 led to a restitution of epithelial characteristics with reciprocal loss of mesenchymal markers. Importantly, TBX2 inhibition abolished tumor cell invasion and the capacity to form lung metastases in a Xenograft mouse model. Meta-analysis of gene expression in over one thousand primary human breast tumors further showed that high TBX2 expression was significantly associated with reduced metastasis-free survival in patients, and with tumor subtypes enriched in EMT gene signatures, consistent with a role of TBX2 in oncogenic EMT. ChIP analysis and cell-based reporter assays further revealed that TBX2 directly represses transcription of E-cadherin, a tumor suppressor gene, whose loss is crucial for malignant tumor progression. Collectively, our results uncover an unanticipated link between TBX2 deregulation in cancer and the acquisition of EMT and invasive features of epithelial tumor cells.


Children with HIV: A scoping review of auditory processing skills.
Auditory processing disorders can negatively affect academic performance in children. They can result from a number of aetiologies, including the human immunodeficiency virus (HIV). Although studies in paediatrics are limited, research suggests that HIV-infected children display poorer auditory processing skills than uninfected children. The aims of this study were to scan the peer-reviewed literature on auditory processing skills in HIV-infected children, to describe how auditory processing was tested, how auditory processing skills were reported, and to identify gaps in current evidence. This systematic scoping review was conducted using a modified version of Arksey and O'Malley's framework. Key words comprised 'HIV', 'auditory processing', 'hearing' and 'child'. Electronic databases were searched for relevant articles published from 1 January 2000 to 30 April 2018, and reference lists of included studies were pearled. Two researchers reviewed the articles and extracted data on sample descriptors, auditory processing testing procedures, and auditory processing skills. A third author collated the results and resolved discrepancies. The American Speech-Language-Hearing Association description of auditory processing skills framed the analysis. Five articles were included in this review (three from Brazil, one each from Mexico and Tanzania). Samples, and methods of testing were heterogeneous. Three studies reported on localization abilities, while gap detection thresholds, performance on dichotic tasks and speech discrimination scores were reported in one article each. No one study tested all areas of auditory processing skills and there was limited information about the auditory processing skills required for learning. This review highlighted the current sparse evidence-base for auditory processing in HIV-infected children. It identified the need to standardise testing procedures, measures of auditory processing skills, and sample selection.


Swine mycoplasmoses.
Mycoplasma hyopneumoniae is the primary agent of enzootic pneumonia in pigs. The lung lesions, generally observed in young pigs, are characterised by a hyperplasia of the epithelial cells and an increased perivascular and peribronchiolar accumulation of mononuclear cells. Following M. hyopneumoniae infection, immune reactions are observed and resistance is induced in pigs. Laboratory diagnosis is generally performed by an immunofluorescent test and by enzyme-linked immunosorbent assay. Antibiotics are useful but the development of resistance has been described. Vaccination seems to be an effective method of controlling the disease. M. hyorhinis, generally transmitted by sows to piglets through nasal secretions, exists in a high percentage in the respiratory tract of healthy pigs. But some strains can induce serofibrinous to fibrinopurulent polyserositis and arthritis. M. hyorhinis is isolated from acute and subacute phase lesions and serum antibodies are detectable. M. hyosynoviae has a special affinity for joint tissue and may cause arthritic disease, leading to economic losses. This mycoplasma is generally located in the tonsils. Piglets are infected by sows after four to six weeks of life. Evidence of disease occurs in animals of between 30 to 40 kg and 100 kg, and bursae and joints are affected. A non-suppurative viscous fluid of a serofibrinous/serosanguineous nature is reported. In chronic cases, the synovial membrane is affected. M. hyosynoviae is isolated from the joints and pharyngeal/tonsillar samples and can induce antibodies in blood and joint fluid. Predisposing factors play an important role. M. flocculare is widely distributed in swine, in normal and pneumonic lungs and in nasal cavities, but no pathogenic capability has been described. There is great interest in this mycoplasma because of the great similarity to M. hyopneumoniae.


Suppressive effect of maslinic acid on PMA-induced protein kinase C in human B-lymphoblastoid cells.
Protein kinase C (PKC) has been implicated in carcinogenesis and displays variable expression profiles during cancer progression. Studies of dietary phytochemicals on cancer signalling pathway regulation have been conducted to search for potent signalling regulatory agents. The present study was designed to evaluate any suppressive effect of maslinic acid on PKC expression in human B-lymphoblastoid cells (Raji cells), and to identify the PKC isoforms expressed. Effects of maslinic acid on PKC activity were determined using a PepTag assay for non-radioactive detection of PKC. The highest expression in Raji cells was obtained at 20 nM PMA induced for 6 hours. Suppressive effects of maslinic acid were compared with those of four PKC inhibitors (H- 7, rottlerin, sphingosine, staurosporine) and two triterpenes (oleanolic acid and ursolic acid). The IC₅₀ values achieved for maslinic acid, staurosporine, H-7, sphingosine, rottlerin, ursolic acid and oleanolic acid were 11.52, 0.011, 0.767, 2.45, 5.46, 27.93 and 39.29 μM, respectively. Four PKC isoforms, PKC βI, βII, δ, and ζ, were identified in Raji cells via western blotting. Maslinic acid suppressed the expression of PKC βI, δ, and ζ in a concentration-dependent manner. These preliminary results suggest promising suppressive effects of maslinic acid on PKC activity in Raji cells. Maslinic acid could be a potent cancer chemopreventive agent that may be involved in regulating many downstream signalling pathways that are activated through PKC receptors.


How well can simulation predict protein folding kinetics and thermodynamics?
Simulation of protein folding has come a long way in five years. Notably, new quantitative comparisons with experiments for small, rapidly folding proteins have become possible. As the only way to validate simulation methodology, this achievement marks a significant advance. Here, we detail these recent achievements and ask whether simulations have indeed rendered quantitative predictions in several areas, including protein folding kinetics, thermodynamics, and physics-based methods for structure prediction. We conclude by looking to the future of such comparisons between simulations and experiments.


Estimation of population bottlenecks during systemic movement of tobacco mosaic virus in tobacco plants.
More often than not, analyses of virus evolution have considered that virus populations are so large that evolution can be explained by purely deterministic models. However, virus populations could have much smaller effective numbers than the huge reported census numbers, and random genetic drift could be important in virus evolution. A reason for this would be population bottlenecks during the virus life cycle. Here we report a quantitative estimate of population bottlenecks during the systemic colonization of tobacco leaves by Tobacco mosaic virus (TMV). Our analysis is based on the experimental estimation of the frequency of different genotypes of TMV in the inoculated leaf, and in systemically infected leaves, of tobacco plants coinoculated with two TMV genotypes. A simple model, based on the probability that a leaf in coinoculated plants is infected by just one genotype and on the frequency of each genotype in the source, was used to estimate the effective number of founders for the populations in each leaf. Results from the analysis of three leaves per plant in plants inoculated with different combinations of three TMV genotypes yielded highly consistent estimates. Founder numbers for each leaf were small, in the order of units. This would result in effective population numbers much smaller than the census numbers and indicates that random effects due to genetic drift should be considered for understanding virus evolution within an infected plant.


Sequence and Structural Diversity of Mouse Y Chromosomes.
Over the 180 My since their origin, the sex chromosomes of mammals have evolved a gene repertoire highly specialized for function in the male germline. The mouse Y chromosome is unique among mammalian Y chromosomes characterized to date in that it is large, gene-rich and euchromatic. Yet, little is known about its diversity in natural populations. Here, we take advantage of published whole-genome sequencing data to survey the diversity of sequence and copy number of sex-linked genes in three subspecies of house mice. Copy number of genes on the repetitive long arm of both sex chromosomes is highly variable, but sequence diversity in nonrepetitive regions is decreased relative to expectations based on autosomes. We use simulations and theory to show that this reduction in sex-linked diversity is incompatible with neutral demographic processes alone, but is consistent with recent positive selection on genes active during spermatogenesis. Our results support the hypothesis that the mouse sex chromosomes are engaged in ongoing intragenomic conflict.


Do the dietary supplements epigallocatechin gallate or vitamin e cause a radiomodifying response on tumors in vivo? A pilot study with murine breast carcinoma.
The primary aim of this study was to evaluate a systematic and reproducible assay to examine the potential radiomodifying effects of vitamin E (VE) or epigallocatechin gallate (EGCG), antioxidants commonly consumed by cancer patients as dietary supplements, on tumor control. C3H mice were randomized to a control diet or to the control diet supplemented with VE or EGCG. A tumor control dose 50% (TCD(50)) assay was used to evaluate for a radiomodifying response in stage IV murine cancer (MCa-IV) tumors, implanted in the hindleg of mice, and allowed to grow to 8 mm before receiving a single dose of radiation. The effects of VE and EGCG on intratumoral angiogenesis and apoptosis were evaluated in a group of nonirradiated mice using immunohistochemical staining. Cell proliferation assays were conducted on MCa-IV tumors in vitro. EGCG slowed tumor growth rate by 10%. EGCG and VE slowed tumor regrowth by 24 to 25%. There were no significant differences in TCD(50) values between the groups (control = 73.9 Gy, VE = 77.2 Gy, EGCG = 76.4 Gy); however, normal tissues were protected from late radiation effects (autoamputations) in the VE group. VE and EGCG increased tumor cell apoptosis and decreased tumor cell proliferation but had no effect on microvessel density. In this pilot study, neither VE nor EGCG exerted a significant radiomodifying effect on the MCa-IV tumor. Nonetheless, the suggestion of a small degree of tumor radioprotection by these antioxidant compounds warrants further research. As supplementation with VE radioprotected normal tissue, additional studies on this putative benefit are recommended.


[Clinical autopsies in Switzerland : A status report].
The number of autopsies has been steadily declining worldwide over the past decades. The reasons for this are diverse. Legislation regarding opposition and consent rules does not appear to have had a significant impact on the autopsy rates. Above all, structural causes and the attitude of the medical profession are the reasons for this decline. The main argument for a high autopsy rate is the identification of diagnostic errors; however, diagnostic discrepancies are relatively independent of the rate of autopsies performed. At the University Hospital (UniversitätsSpital) Zurich it could be shown in a study that from 1972-2002 the frequency of relevant diagnostic discrepancies (classes I and II) decreased from 30% to 7%. The aim of this article is to present the necessity of a stable autopsy rate and to examine the situation of the autopsy in Switzerland. For this purpose, the importance of autopsies in the fields of quality assurance of medical diagnostics, cancer statistics, medical research as well as further education of doctors in Switzerland is shown. Efforts are being made by the pathologists to counteract the declining autopsy rates. Declining autopsy numbers have a significant influence on cancer statistics. The rate of newly discovered tumors in autopsies in Switzerland decreased from 42% in 1980 to 17% in 2010. Pediatric autopsies are an important tool for quality assurance of medical diagnostics in neonatology and pediatrics in Switzerland, but the rate of autopsies carried out is also declining. Postmortem magnetic resonance imaging (MRI) examinations (virtopsy) could increase the acceptance of the parents for an autopsy in the future. Autopsies make an important contribution in research and in documentation of therapy-associated side effects and they are an important component of further education of the upcoming medical generations.


Isolation of Bacteriophages for Clinically Relevant Bacteria.
The isolation of bacteriophages targeting most clinically relevant bacteria is reasonably straightforward as long as its targeted host does not have complex chemical, physical, and environmental requirements. Often, sewage, soil, feces, and different body fluids are used for bacteriophage isolation procedures, and following enrichment, it is common to obtain more than a single phage in a sample. This chapter describes a simple method for the enrichment and isolation of bacteriophages from liquid and solid samples that can be adapted for different clinically important aerobic bacteria.


Phosphorylation of dipteran chromosomes and rat liver nuclei.
Side-chain modifications of the nucleoproteins are believed to be involved in the control of gene function. Rat liver nuclei and chromatin incubated in adenosine triphosphate labeled in the gamma position with phosphorus-32 demonstrated rapid phosphorylation in vitro of the nuclear proteins. Dipteran salivary glands incubated in either labeled adenosine triphosphate or ortho-phosphate labeled with phosphorus-32 showed that there is phosphorylation of chromosomal protein. The phosphorus is associated in protein from both liver and salivary gland nuclei predominantly with phosphoserine.


In vivo noninvasive measurement of blood glucose by near-infrared diffuse-reflectance spectroscopy.
This paper reports in situ noninvasive blood glucose monitoring by use of near-infrared (NIR) diffuse-reflectance spectroscopy. The NIR spectra of the human forearm were measured in vivo by using a pair of source and detector optical fibers separated by a distance of 0.65 mm on the skin surface. This optical geometry enables the selective measurement of dermis tissue spectra due to the skin's optical properties and reduces the interference noise arising from the stratum corneum. Oral glucose intake experiments were performed with six subjects (including a single subject with type I diabetes) whose NIR skin spectra were measured at the forearm. Partial least-squares regression (PLSR) analysis was carried out and calibration equations were obtained with each subject individually. Without exception among the six subjects, the regression coefficient vectors of their calibration models were similar to each other and had a positive peak at around 1600 nm, corresponding to the characteristic absorption peak of glucose. This result indicates that there is every possibility of glucose detection in skin tissue using our measurement system. We also found that there was a good correlation between the optically predicted values and the directly measured values of blood samples with individual subjects. The potential of noninvasive blood glucose monitoring using our methodology was demonstrated by the present study.


Phosphorus flux from bottom sediments in Lake Eucha, Oklahoma.
Phosphorus inputs into reservoirs include external sources from the watershed and internal sources from the reservoir bottom sediments. This study quantified sediment P flux in Lake Eucha, northeastern Oklahoma, USA, and evaluated the effectiveness of chemical treatment to reduce sediment P flux. Six intact sediment-water columns were collected from three sites in Lake Eucha near the reservoir channel at depths of 10 to 15 m. Three intact sediment and water columns from each site were incubated for 21 d at approximately 22 degrees C under aerobic conditions, and three were incubated under anaerobic conditions (N2 with 300 ppm CO2); sediment P flux was estimated over the 21 d for each core. The overlying water in the cores was bubbled with air for approximately 1 wk and then treated with aluminum sulfate (alum). The cores were incubated at approximately 22 degrees C for an additional 14 d under aerobic or anaerobic conditions, and sediment P flux after alum treatment was estimated for each core. Sediment P flux was approximately four times greater under anaerobic conditions compared with aerobic conditions. Alum treatment of the intact sediment-water columns reduced (8x) sediment P flux under anaerobic conditions. Internal P flux (1.03 and 4.40 mg m(-2) d(-1) under aerobic and anaerobic conditions, respectively) was greater than external P flux (0.13 mg m(-2) d(-1)). The internal P load (12 Mg yr(-1)) from reservoir bottom sediments was almost 25% of the external P load (approximately 48 Mg yr(-1)) estimated using a calibrated watershed model.


Time-resolved ultrastructure of the cortical actin cytoskeleton in dynamic membrane blebs.
Membrane blebbing accompanies various cellular processes, including cytokinesis, apoptosis, and cell migration, especially invasive migration of cancer cells. Blebs are extruded by intracellular pressure and are initially cytoskeleton-free, but they subsequently assemble the cytoskeleton, which can drive bleb retraction. Despite increasing appreciation of physiological significance of blebbing, the molecular and, especially, structural mechanisms controlling bleb dynamics are incompletely understood. We induced membrane blebbing in human HT1080 fibrosarcoma cells by inhibiting the Arp2/3 complex. Using correlative platinum replica electron microscopy, we characterize cytoskeletal architecture of the actin cortex in cells during initiation of blebbing and in blebs at different stages of their expansion-retraction cycle. The transition to blebbing in these conditions occurred through an intermediate filopodial stage, whereas bleb initiation was biased toward filopodial bases, where the cytoskeleton exhibited local weaknesses. Different stages of the bleb life cycle (expansion, pausing, and retraction) are characterized by specific features of cytoskeleton organization that provide implications about mechanisms of cytoskeleton assembly and bleb retraction.


[Characteristics of violence during teenage pregnancy in Lima, Peru].
To determine the characteristics of violence seen in pregnant teenagers who were treated at the Instituto Nacional Materno Perinatal (INMP) in Lima, Peru. A cross-sectional study was carried out by INMP between January and March, 2010 using a probabilistic and systematic sampling. The study unit comprises every hospitalized teenager who had just given birth and who lived in Lima. A semi-structured interview was conducted. History of violence was operationalized into: verbal violence (insults, ridicule, and humiliation), physical violence (arm pulling, hair pulling, pushes), direct aggression (slaps, kicking, burns) and sexual violence (sexual intercourse without consent). 292 teenage mothers aged 16,5 ± 1 in average took part in the study. 47.9% lived with their partners and 51.4% were single. In 97.3% of the cases, they got pregnant as a result of a conserted sexual relationship, while 2.7% got pregnant as a result of rape. 90.1% of teenage mothers reported not having planned the pregnancy. Conserning history of violence: 48.1% had had verbal violence, 17.1% physical violence, 8.2% direct aggression and 6.8% sexual violence. Violence during teenage pregnancy is not an isolated event; actually, it is rather common in any of its forms.


Migration of Lebanese nurses: a questionnaire survey and secondary data analysis.
Nursing is becoming a mobile profession. Nurse migration is multifactorial and not limited to financial incentives. Non-economic factors that might lead to migration include poor recruitment and retention strategies, poor job satisfaction and working conditions, socio-political and economic stability, and the poor social image of the nursing profession. Lebanon is facing a problem of excessive nurse migration to countries of the Gulf, North America and Europe. No study has been conducted to understand the determinants and magnitude of the problem. The objective of this study is to provide an evidence base for understanding the incidence of nurse migration out of Lebanon, its magnitude and reasons. A cross-sectional research design comprising both quantitative and qualitative methods was employed to achieve the stated objectives. This includes a survey of nursing schools in Lebanon, survey of nurse recruitment agencies, secondary data analysis and survey of migrant nurses. An estimated one in five nurses that receive a bachelors of science in nursing migrates out of Lebanon within 1 or 2 years of graduation. The majority of nurses migrate to countries of the Gulf. The main reasons for migration included: shift work, high patient/nurse ratios, lack of autonomy in decision-making, lack of a supportive environment, and poor commitment to excellent nursing care. Further, nurses reported that combinations of financial and non-financial incentives can encourage them to return to practice in Lebanon. The most recurring incentives (pull factors) to encourage nurses to return to practice in Lebanon included educational support, managerial support, better working conditions, utilization of best nursing practices and autonomy. Nurse migration and retention have become major health workforce issues confronting many health systems in the East Mediterranean Region. Our study demonstrated that nurse migration is a product of poor management and lack of effective retention strategies and sufficient knowledge about the context, needs and challenges facing nurses. Nurse migration in Lebanon underscores the importance of developing a monitoring system that would identify implications and help implement innovative retention strategies. Nurse migration out of Lebanon is likely to persist and even increase if underlying factors are not properly resolved.


Effects of stomatal delays on the economics of leaf gas exchange under intermittent light regimes.
• Understory plants are subjected to highly intermittent light availability and their leaf gas exchanges are mediated by delayed responses of stomata and leaf biochemistry to light fluctuations. In this article, the patterns in stomatal delays across biomes and plant functional types were studied and their effects on leaf carbon gains and water losses were quantified. • A database of more than 60 published datasets on stomatal responses to light fluctuations was assembled. To interpret these experimental observations, a leaf gas exchange model was developed and coupled to a novel formulation of stomatal movement energetics. The model was used to test whether stomatal delays optimize light capture for photosynthesis, whilst limiting transpiration and carbon costs for stomatal movement. • The data analysis showed that stomatal opening and closing delays occurred over a limited range of values and were strongly correlated. Plant functional type and climate were the most important drivers of stomatal delays, with faster responses in graminoids and species from dry climates. • Although perfectly tracking stomata would maximize photosynthesis and minimize transpiration at the expense of large opening costs, the observed combinations of opening and closure times appeared to be consistent with a near-optimal balance of carbon gain, water loss and movement costs.


A predictable comeback: the second pandemic of infections caused by Neisseria meningitidis serogroup A subgroup III in Africa, 1995.
Between 14 January and 4 April 1995 we isolated and characterized 44 meningococcal strains in Cameroon, Chad, Niger, and Burkina Faso; among these was the strain A:4:P1.9/clone III-1, which was involved in the second meningitis pandemic. This isolate was found in the clonal form in Niger and strains of the ET-37 complex were also found in the other three study countries, but apparently did not cause epidemics. One strain (Y:2a:P1.2,5 (ET-37 complex)) was isolated in January 1995 and another (A:4:P1.9) in March 1995 in Garoua (Cameroon). Eight strains were isolated in Moundou (Chad) between January and April 1995: the A:4:P1.9/clone III-1 (1 strain); members of the ET-37 complex (Y:2a:P1.2,5 (4 strains), Y:NT:P1.2,5 (1 strain), and Y:2a:-(1 strain)); and serogroup X (1 strain). In Niger, 31 strains were isolated between February and April 1995 from different regions. All were A:4:P1.9/clone III-1; between November 1994 and April 1995 there were 23814 cases of meningitis reported of which 2227 resulted in death. Three strains were isolated in Burkina Faso in April 1995: two were Y:2a:P1.2,5 (ET-37 complex) and one was A:4:P1.9/clone III-1. Thus in 1995 the epidemic and invasive strain (A:4:P1.9/clone III-1) responsible for the second pandemic was present in the four countries (Cameroon, Chad, Niger and Burkino Faso) that make up the area frequently affected by such epidemics and where cases are generally reported during the dry season.


RNA interference-based therapy for spinocerebellar ataxia type 7 retinal degeneration.
Spinocerebellar ataxia type 7 (SCA7) is an autosomal dominant neurodegenerative disease characterized by loss of motor coordination and retinal degeneration with no current therapies in the clinic. The causative mutation is an expanded CAG repeat in the ataxin-7 gene whose mutant protein product causes cerebellar and brainstem degeneration and retinal cone-rod dystrophy. Here, we reduced the expression of both mutant and wildtype ataxin-7 in the SCA7 mouse retina by RNA interference and evaluated retinal function 23 weeks post injection. We observed a preservation of normal retinal function and no adverse toxicity with ≥50% reduction of mutant and wildtype ataxin-7 alleles. These studies address an important safety concern regarding non-allele specific silencing of ataxin-7 for SCA7 retinal therapy.


Identification of a signal transduction switch in the chemokine receptor CXCR1.
Chemokine receptors belong to the superfamily of G protein-coupled receptors, which regulate the trafficking and activation of leukocytes, and operate as coreceptors in the entry of HIV-1. To investigate the early steps in the signal transmission from the chemokine-binding site to the G protein-coupling region we engineered metal ion-binding sites at putative extracellular sites in the chemokine receptor CXCR1. We introduced histidines into sites located in the second and third putative extracellular loops of CXCR1, creating single, double, and triple mutant receptors: R199H, R203H, D265H, R199H/R203H, R199H/D265H, R203H/D265H, R203H/H207Q, and R199H/R203H/D265H. Cells expressing the double mutants R199H/D265H and R203H/D265H and the triple mutant R199H/R203H/D265H failed to trigger interleukin 8-dependent calcium responses. Interestingly, calcium responses mediated by the single mutant R203H and the double mutants R199H/R203H and R203H/H207Q were blocked by Zn(II), indicating the creation of a functional metal ion-binding site. On the other hand, cells expressing all single, double, or triple histidine-substituted CXCR1 demonstrated high affinity binding to interleukin 8 in the presence and absence of metal ions. These findings indicate that occupation of the engineered metal-binding site uncouples the chemokine-binding site from the activation mechanism in CXCR1. Most importantly, we identify for the first time elements of an early signal transduction switch of chemokine receptors before the activation of cytoplasmic G proteins.


Redox-Active Sensing by Bacterial DksA Transcription Factors Is Determined by Cysteine and Zinc Content.
The four-cysteine zinc finger motif of the bacterial RNA polymerase regulator DksA is essential for protein structure, canonical control of the stringent response to nutritional limitation, and thiol-based sensing of oxidative and nitrosative stress. This interdependent relationship has limited our understanding of DksA-mediated functions in bacterial pathogenesis. Here, we have addressed this challenge by complementing ΔdksA Salmonella with Pseudomonas aeruginosa dksA paralogues that encode proteins differing in cysteine and zinc content. We find that four-cysteine, zinc-bound (C4) and two-cysteine, zinc-free (C2) DksA proteins are able to mediate appropriate stringent control in Salmonella and that thiol-based sensing of reactive species is conserved among C2 and C4 orthologues. However, variations in cysteine and zinc content determine the threshold at which individual DksA proteins sense and respond to reactive species. In particular, zinc acts as an antioxidant, dampening cysteine reactivity and raising the threshold of posttranslational thiol modification with reactive species. Consequently, C2 DksA triggers transcriptional responses in Salmonella at levels of oxidative or nitrosative stress normally tolerated by Salmonella expressing C4 orthologues. Inappropriate transcriptional regulation by C2 DksA increases the susceptibility of Salmonella to the antimicrobial effects of hydrogen peroxide and nitric oxide, and attenuates virulence in macrophages and mice. Our findings suggest that the redox-active sensory function of DksA proteins is finely tuned to optimize bacterial fitness according to the levels of oxidative and nitrosative stress encountered by bacterial species in their natural and host environments. In order to cause disease, pathogenic bacteria must rapidly sense and respond to antimicrobial pressures encountered within the host. Prominent among these stresses, and of particular relevance to intracellular pathogens such as Salmonella, are nutritional restriction and the enzymatic generation of reactive oxygen and nitrogen species. The conserved transcriptional regulator DksA controls adaptive responses to nutritional limitation, as well as to oxidative and nitrosative stress. Here, we demonstrate that each of these functions contributes to bacterial pathogenesis. Our observations highlight the importance of metabolic adaptation in bacterial pathogenesis and show the mechanism by which DksA orthologues are optimized to sense the levels of oxidative and nitrosative stress encountered in their natural habitats. An improved understanding of the conserved processes used by bacteria to sense, respond to, and limit host defense will inform the development of novel strategies to treat infections caused by pathogenic, potentially multidrug-resistant bacteria.


Chronic exertional compartment syndrome in diabetes mellitus.
Intermittent claudication is common in diabetes mellitus and usually (attributed to arterial disease) . However, a proportion of patients with diabetes have symptoms of claudication without signs of vascular disease and these patients were evaluated for chronic exertional compartment syndrome. Forty-two patients with diabetes (10 men, 32 women), earlier investigated at diabetic clinics because of claudication with no explanation for the symptoms, were examined. Their median age was 48 years (18-72 years) and the median duration of diabetes was 29 years (1-45 years). Thirty-one patients had Type 1 diabetes, 11 had Type 2 diabetes and 29 had diabetic complications. All were investigated clinically, with radiography, bone scan and intramuscular pressure measurements. Thirty-eight of 42 patients with diabetes were diagnosed with chronic exertional compartment syndrome of the lower leg and 32 were treated surgically. Thirty-one patients were operated with fasciotomy of the anterior compartment and 18 also with fasciotomy of the posterior compartment. Additionally, one patient had only fasciotomy of the posterior compartment. Fourteen of 32 surgically treated patients (27 legs) were followed for more than 2 years and rated the post-operative outcome as excellent or good in 21 of the treated legs. The walking distance before lower leg pain increased in all but one patient and seven patients reported unrestricted walking ability. Chronic exertional compartment syndrome should be considered as a differential diagnoses in patients with diabetes and exercise-related lower leg pain. The results after surgery are encouraging and the increased walking ability is beneficial in the treatment of diabetes.


NACore Amyloid Formation in the Presence of Phospholipids.
Amyloids are implicated in many diseases, and disruption of lipid membrane structures is considered as one possible mechanism of pathology. In this paper we investigate interactions between an aggregating peptide and phospholipid membranes, focusing on the nanometer-scale structures of the aggregates formed, as well as on the effect on the aggregation process. As a model system, we use the small amyloid-forming peptide named NACore, which is a fragment of the central region of the protein α-synuclein that is associated with Parkinson's disease. We find that phospholipid vesicles readily associate with the amyloid fibril network in the form of highly distorted and trapped vesicles that also may wet the surface of the fibrils. This effect is most pronounced for model lipid systems containing only zwitterionic lipids. Fibrillation is found to be retarded by the presence of the vesicles. At the resolution of our measurements, which are based mainly on cryogenic transmission electron microscopy (cryo-TEM), X-ray scattering, and circular dichroism (CD) spectroscopy, we find that the resulting aggregates can be well fitted as linear combinations of peptide fibrils and phospholipid bilayers. There are no detectable effects on the cross-β packing of the peptide molecules in the fibrils, or on the thickness of the phospholipid bilayers. This suggests that while the peptide fibrils and lipid bilayers readily co-assemble on large length-scales, most of them still retain their separate structural identities on molecular length-scales. Comparison between this relatively simple model system and other amyloid systems might help distinguish aspects of amyloid-lipid interactions that are generic from aspects that are more protein specific. Finally, we briefly consider possible implications of the obtained results for in-vivo amyloid toxicity.


A new slant to the Z ring and bacterial cell branch formation.
Rod-shaped bacteria such as Escherichia coli accurately maintain their shape from generation to generation. The cytoskeletal proteins MreB and FtsZ, which respectively guide parallel growth of the sidewall and perpendicular growth of the division septum, are important to maintain a straight sidewall and uniformly rounded cell poles. FtsZ normally assembles into a ring at the cell midpoint, called the Z ring, which is oriented perpendicular to the cell's axis and is thus in perfect position to guide growth of a perpendicular septum. In this issue of Molecular Microbiology, Potluri et al. show that low molecular weight penicillin binding proteins, particularly PBP5, have a role in maintaining the perpendicular geometry of the Z ring and subsequent septum in E. coli. When these factors are absent or perturbed, division septa are readily deformed, which results in abnormal cell poles that often bifurcate over time to generate branches. The data suggest that cellular branching in E. coli is specifically induced by aberrant septation events caused by mis-oriented Z rings and not by deformation of a growing cell pole or emergence of new tips from the sidewall, which are likely mechanisms of branching in other bacterial families.


Congenital myasthenic syndrome: a case report.
Congenital myasthenic syndromes (CMS) are diseases of the neuromuscular junction. They usually belong to the disease groups that begin in the infantile or childhood period and carry genetic characteristics. The following is important in establishing the diagnosis of this disease: clinical findings, electromyography, genetic tests, determination of serum acetylcholine receptor antibodies. Acetylcholine esterase inhibitor drugs are used in treatment of CMS. A seven-month old male patient was brought to our department with the complaints of difficult breathing, falling of the eyelids and swallowing difficulty. With clinical and laboratory findings, he was diagnosed with congenital myasthenia and treatment was started. CMS should be suspected in patients with no pathological findings on the physical examination, and normal chest X-rays.


[Characteristics of postoperative period in children with funnel chest deformity after thoracoplasty].
The efficiency of postoperative analgesia using the epidural infusion of bupivacaine (0.2 mg/kg/hr) in 42 children and with opioid analgesics (1 mg per year of life) was studied in 17 children after Paltia thoracoplasty for funnel chest deformity. Analysis of hemodynamic and external respiratory parameters and visual pain scores demonstrated the high postoperative efficiency of analgesia using the epidural infusion of bupivacaine. By causing respiratory depression, opioid analgesics did not deteriorate alveolar ventilation during active postoperative management.


Pharmacokinetics and pharmacodynamics of the myotoxic venom of Pseudechis australis (mulga snake) in the anesthetised rat.
Myotoxicity is a common clinical effect of snake envenoming and results from either local or systemic myotoxins in snake venoms. Although numerous myotoxins have been isolated from snake venoms, there has been limited study on the relationship between the time course of venom concentrations (pharmacokinetics) and the time course of muscle injury measured as a rise in creatine kinase (CK) (pharmacodynamics). The aim of this study was to develop an in vivo model of myotoxicity to investigate the time course of myotoxicity and the effect of antivenom. Anesthetised rats were administered Pseudechis australis (mulga snake) venom either through i.v., i.m. or s.d. route, including a range of doses (5-100 μg/kg). Serial blood samples were collected for measurement of venom using enzyme immunoassay and measurement of CK and creatinine. Antivenom was administered before, 1 and 6 h after venom administration to investigate its effect on muscle injury. Plots of venom and CK versus time were made and the area under the curve (AUC) was calculated. There was a significant dose-dependent increase in CK concentration after administration of P. australis venom, which was greatest for i.v. administration. Timed measurement of venom concentrations showed a rapid absorption through s.d. and i.m. routes and a delayed rise in CK concentrations following any route. Antivenom prevented myotoxicity shown by a decrease in the CK AUC, which was most effective if given earliest. There was a rise in creatinine following i.v. venom administration. The study shows the delayed relationship between venom absorption and the rise in CK, consistent with the delayed onset of myotoxicity in human envenoming. Antivenom prevented myotoxicity more effectively if given earlier.


Sulphur dioxide suppresses inflammatory response by sulphenylating NF-κB p65 at Cys38 in a rat model of acute lung injury.
The present study was designed to investigate whether endogenous sulphur dioxide (SO2) controlled pulmonary inflammation in a rat model of oleic acid (OA)-induced acute lung injury (ALI). In this model, adenovirus expressing aspartate aminotransferase (AAT) 1 was delivered to the lungs, and the levels of SO2 and proinflammatory cytokines in rat lung tissues were measured. In the human alveolar epithelial cell line A549, the nuclear translocation and DNA binding activities of wild-type (wt) and C38S (cysteine-to-serine mutation at p65 Cys38) NF-κB p65 were detected. GFP-tagged C38S p65 was purified from HEK 293 cells and the sulphenylation of NF-κB p65 was studied. OA caused a reduction in SO2/AAT pathway activity but increased pulmonary inflammation and ALI. However, either the presence of SO2 donor, a combination of Na2SO3 and NaHSO3, or AAT1 overexpression in vivo successfully blocked OA-induced pulmonary NF-κB p65 phosphorylation and consequent inflammation and ALI. Either treatment with an SO2 donor or overexpression of AAT1 down-regulated OA-induced p65 activity, but AAT1 knockdown in alveolar epithelial cells mimicked OA-induced p65 phosphorylation and inflammation in vitro Mechanistically, OA promoted NF-κB nuclear translocation, DNA binding activity, recruitment to the intercellular cell adhesion molecule (ICAM)-1 promoter, and consequent inflammation in epithelial cells; these activities were reduced in the presence of an SO2 donor. Furthermore, SO2 induced sulphenylation of p65, which was blocked by the C38S mutation on p65 in epithelial cells. Hence, down-regulation of SO2/AAT is involved in pulmonary inflammation during ALI. Furthermore, SO2 suppressed inflammation by sulphenylating NF-κB p65 at Cys38.


Electronic excitation and thermal effects in alkali-halide cluster anions
We have observed electronically excited states in alkali-halide cluster anions with one excess electron. Using photoelectron spectroscopy, we have found two narrow states in (KI)-2, (NaI)-2, and (NaCl)-2, consistent with a dipole-bound electron, while larger cluster anions exhibit a single broad excited state. In the larger systems, electronic excitation is often accompanied by vibrational excitation and thus a change in cluster temperature. Such temperature changes affect cluster structure and in some cases lead to rapid thermal isomerization.


Totally implantable central venous catheters of the port-a-cath type: complications due to its use in the treatment of cancer patients.
A multidisciplinary approach to the treatment of patients with malignant diseases requires adequate venous access in order to safely administer chemotherapy, blood transfusion and blood products, antibiotics, rehydratation and total parenteral nutrition. The insertion of the central venous catheter (CVC), its use and its maintenance can be accompanied by multiple complications. Fifty cancer patients were retrospectively enrolled in this study. The obligatory inclusion criterion was an implanted CVC of the port-a-cath type, inserted for chemotherapy administration. This study included patients who had their catheters inserted in the period from 2001 to 2012. The median patient age was 44 years (range 28- 68). Thirty five patients (70%) were female and 15 (30%) male. The port-a-cath had been used from 1 to 40 months (16.8 ± 9 months on average). Breast cancer was the most frequent malignancy (18 patients, 36%). The overall incidence of reported complications was 38%. The most common complications were infections and thromboembolic events, each with an incidence of 10 %. The malposition and disconnection of the port-a-cath were in second place, each with an incidence of 6%. Insertion of the CVC carries the possibility of serious complications (thrombosis, infections, occlusions). However, correct implantation and handling performed by experienced and trained surgical and other medical staff significantly decrease the incidence of these complications. The use of the CVC has greatly improved the quality of life and also decreased the morbidity and mortality of the cancer patients in our study.


The activation domain of a hormone inducible HTLV-1 Rex protein determines colocalization with the nuclear pore.
Human T-cell leukemia/lymphoma virus type 1 (HTLV-1) Rex is an essential regulatory protein that acts at the posttranscriptional level to promote expression of unspliced and singly spliced genes of the virus. Rex functions have been attributed to at least three separate domains of the protein determining nuclear/nucleolar accumulation and RNA binding (overlapping), multimerization, and nuclear export of Rex-responsive RNA. The steady-state intracellular localization of functional Rex molecules is mainly nucleolar. Fusions of wild-type Rex and the ligand binding domain of human estrogen receptor (ER) produced conditional molecules (ERRex and ERalaRex), which remained cytoplasmic in the absence of hormone and in response to hormone colocalized with the nuclear pore complex (NPC). These molecules induced in a hormone-dependent manner the expression of a Rex reporter plasmid and of the HTLV-1 Env protein and fusion of Env expressing cells. In contrast, activation domain mutants (ERRex delta and ERRexGly) translocated from the cytoplasm and acquired a diffuse nuclear localization. These mutants did not associate with the NPC and failed to show any of the expected Rex functions. Rex functions were perturbed by inactivating the RNA binding domain (mutant ERM2) or the oligomerization domain (mutant ERM7). However, these two mutant fusion proteins exhibited a hormone-dependent NPC colocalization. These observations provide in vivo evidence that intranuclear translocation of intact Rex to the NPC is dependent exclusively on a functional activation domain and is not influenced by binding to the target RNA.


Delays in diagnosis and treatment among children and adolescents with cancer in Canada.
Few studies have investigated delays in diagnosis and treatment among children and adolescents with cancer, especially from the perspective of an entire country. Detailed understanding of delays along the continuum of cancer patient care is important in order to establish appropriate benchmarks for timely oncological care. Our objective was to characterise the different components of delay in 2,896 Canadian children and adolescents (aged 0-19 years) with cancer that were enrolled in the Treatment and Outcome Surveillance component of the Canadian Childhood Cancer Surveillance and Control Program from 1995 to 2000. We examined median and standardised means concerning the distribution of delay times across categories of pertinent variables and over time. The word "delay" was used simply to represent a time interval, measured in days, without implying whether this interval exceeded a particular threshold of clinical acceptability. The median times (and inter-quartile ranges) for patient, diagnosis and healthcare system delays for all cancers were 9 (1-31), 30 (13-69) and 12 (4-35) days, respectively. The median total delay was 34 (16-76) days. Patient and referral delays were the longest time segments influencing timely diagnosis. Differences in delays were observed across age groups, cancer types and geographical regions. There was a significant trend for decreasing delays to diagnosis and treatment.


Success of Senegal's first nationwide distribution of long-lasting insecticide-treated nets to children under five - contribution toward universal coverage.
In 2009, the first national long-lasting insecticide-treated net (LLIN) distribution campaign in Senegal resulted in the distribution of 2.2 million LLINs in two phases to children aged 6-59 months. Door-to-door teams visited all households to administer vitamin A and mebendazole, and to give a coupon to redeem later for an LLIN. A nationwide community-based two-stage cluster survey was conducted, with clusters selected within regions by probability proportional to size sampling, followed by GPS-assisted mapping, simple random selection of households in each cluster, and administration of a questionnaire using personal digital assistants (PDAs). The questionnaire followed the Malaria Indicator Survey format, with rosters of household members and bed nets, and questions on campaign participation. There were 3,280 households in 112 clusters representing 33,993 people. Most (92.1%) guardians of eligible children had heard about the campaign, the primary sources being health workers (33.7%), neighbours (26.2%), and radio (22.0%). Of eligible children, 82.4% received mebendazole, 83.8% received vitamin A, and 75.4% received LLINs. Almost all (91.4%) LLINs received during the campaign remained in the household; of those not remaining, 74.4% had been given away and none were reported sold. At least one insecticide-treated net (ITN) was present in 82.3% of all households, 89.2% of households with a child < 5 years and 57.5% of households without a child < 5 years. Just over half (52.4%) of ITNs had been received during the campaign. Considering possible indicators of universal coverage, 39.8% of households owned at least one ITN per two people, 21.6% owned at least one ITN per sleeping space and 34.7% of the general population slept under an ITN the night before the survey. In addition, 45.6% of children < 5 years, and 49.2% of pregnant women had slept under an ITN. The nationwide integrated LLIN distribution campaign allowed household ITN ownership of one or more ITNs to surpass the RBM target of 80% set for 2010, though additional distribution strategies are needed to reach populations missed by the targeted campaign and to reach the universal coverage targets of one ITN per sleeping space and 80% of the population using an ITN.


[The pathologic urinary bladder in the sonographic image].
Intracavitary sonography proved to be advantageous with regard to the T-classification of urinary bladder tumours in comparison with conventional diagnostic methods. Besides the good practicability, the high degree of correspondence between sonography and histopathological findings is emphasized. In 97 patients with urothelial bladder cancer this method yielded 63% correct results. In 32% overstaging was noticed, possible factors of which were exemplified. Part of the sonographic failures can be avoided by means of technical manipulation (gain-modification) together with increasing experience of the examiner. In particular benign bladder disease, which can be sonographically confused with cancer, has to be considered. These cases indicate the need to correlate particular sonographic findings with histopathology.


Comparison of the Panther Fusion and Allplex assays for the detection of respiratory viruses in clinical samples.
Respiratory viral infections are the most frequent clinical syndrome affecting both children and adults, and early detection is fundamental to avoid infection-related risks and reduce the healthcare costs incurred by unnecessary antibiotic treatments. In this study, performance characteristics of two commercial methods, the Panther Fusion® assay (Hologic Inc., San Diego, CA, USA) were compared to Allplex™ respiratory panels (Seegene, Seoul, South Korea) for the detection of influenza A (Flu A), influenza B (Flu B), respiratory syncytial virus (RSV), parainfluenza virus (PIV), human metapneumovirus (hMPV), rhinovirus (RV) and adenovirus (AdV) targets. A total of 865 specimens collected prospectively and retrospectively were included, and discordant results were further examined using another commercial product, R-GENE™ respiratory kits (bioMérieux, Marcy l'Etoile, France). There was high agreement between both methods, with 98.6% concordance and a kappa (k) value of 0.9 (95% CI: 0.89-0.92). A specific analysis of both methods for each viral agent demonstrated comparable sensitivity and specificity, both ranging from 0.83 to 1 with good predictive values for the prospective part of the study. Good agreement between both methods was also found for the κ values obtained (ranging from 97.55% to 98.9%), with the lowest for hMPV (k = 0.83, 95% CI: 0.75-0.91) and RV (k = 0.73, 95% CI: 0.65-0.81). Amplification efficiency, measured according to the value of the cycle threshold (Ct) obtained in each of the amplifications in both tests, was significantly better with Panther Fusion for Flu A, Flu B, hMPV and RV. Regarding discordant results, R-GENE showed higher agreement with Panther Fusion-positive specimens (negative for Allplex; n = 28/71, 34.9%) than with Allplex-positive samples (negative for Panther Fusion; n = 7/49, 14.3%). In summary, Panther Fusion proved to be a more efficient fully-automated methodology, requiring shorter hands-on and turnaround times than Allplex.


The Impact of Bleeding Avoidance Strategies on Hospital-Level Variation in Bleeding Rates Following Percutaneous Coronary Intervention: Insights From the National Cardiovascular Data Registry CathPCI Registry.
The aim of this study was to explore whether the use of bleeding avoidance strategies (BAS) explains variability in hospital-level bleeding following percutaneous coronary intervention. Prior studies have reported that bleeding rates following percutaneous coronary intervention vary markedly among hospitals, but the extent to which use of BAS explains this variation is unknown. Using the American College of Cardiology National Cardiovascular Data Registry's CathPCI Registry, estimated hospital-level bleeding rates from 2,459,686 procedures at 1,358 sites were determined. A series of models were fit to estimate random-effect variance, adjusting for patient risk (using the validated CathPCI bleeding risk model, C statistic = 0.77) and various combinations of BAS (transradial access, bivalirudin, vascular closure device use). The rate of any BAS use was also estimated for each hospital, and the association between percentage BAS use and predicted bleeding rates was determined. In total, 125,361 bleeding events (5.1%) were observed; patients experiencing bleeding events had lower rates of radial access (5.0% vs. 11.2%; p < 0.001), bivalirudin therapy (43.8% vs. 59.4%), and vascular closure device use (32.9% vs. 42.4%, p < 0.001) than those without bleeding. There was significant variation in bleeding rates across hospitals (median 5.0%; interquartile range [IQR]: 2.7% to 6.6%), which persisted after incorporating patient-level risk (median 5.1%; IQR: 4.0% to 4.4%). Patient factors accounted for 20% of the overall hospital-level variation, and radial access plus bivalirudin use accounted for an additional 7.8% of the overall hospital-level variation. The median hospital rate of any BAS use was 86.6% (IQR: 72.5% to 94.1%). A significant decrease in observed hospital-level bleeding was seen in hospitals above the median in BAS use (adjusted odds ratio: 0.90; 95% confidence interval: 0.88 to 0.93). A modest proportion of the variation in hospitals' rates of bleeding following percutaneous coronary intervention is attributable to differential use of BAS. Further analyses are required to determine the remaining approximately 70% causes of variation in percutaneous coronary intervention bleeding seen among hospitals.


Understanding circadian regulation of carbohydrate metabolism in Arabidopsis using mathematical models.
C3 plants assimilate carbon by photosynthesis only during the day, but carbon resources are also required for growth and maintenance at night. To avoid carbon starvation, many plants store a part of photosynthetic carbon in starch during the day, and degrade it to supply sugars for growth at night. In Arabidopsis, starch accumulation in the day and degradation at night occur almost linearly, with the shape of this diel starch profile adaptively changing to allow continuous supply of sugar even in long-night conditions. The anticipation of dawn required to ensure linear consumption of starch to almost zero at dawn presumably requires the circadian clock. We review the links between carbon metabolism and the circadian clock, and mathematical models aimed at explaining the diel starch profile. These models can be considered in two classes, those that assume the level of available starch is sensed and the system ensures linearity of starch availability, and those in which sugar sensing is assumed, yielding linearity of starch availability as an emergent property of sucrose homeostasis. In the second class of model the feedback from starch metabolism to the circadian clock is considered to be essential for adaptive response to diverse photoperiods, consistent with recent empirical data demonstrating entrainment of the circadian clock by photosynthesis. Knowledge concerning the mechanisms regulating the dynamics of starch metabolism and sugar homeostasis in plants is required to develop new theories about the limitations of growth and biomass accumulation.


A Contemporary Overview of PPARα/γ Dual Agonists for the Management of Diabetic Dyslipidemia.
Diabetes mellitus and concomitant dyslipidemia, being referred to as 'diabetic dyslipidemia', are the foremost detrimental factors documented to play a pivotal role in cardiovascular illness. Diabetic dyslipidemia is associated with insulin resistance, high plasma triglyceride levels, low HDL-cholesterol concentration and elevated small dense LDL-cholesterol particles. Maintaining an optimal glucose and lipid levels in patients afflicted with diabetic dyslipidemia could be a major task that might require a well-planned diet-management system and regular physical activity, or otherwise an intake of combined antidiabetic and antihyperlipidemic medications. Synchronized treatment which efficiently controls insulin resistance-associated diabetes mellitus and co-existing dyslipidemia could indeed be a fascinating therapeutic option in the management of diabetic dyslipidemia. Peroxisome proliferator-activated receptors α/γ (PPARα/γ) dual agonists are such kind of drugs which possess therapeutic potentials to treat diabetic dyslipidemia. Nevertheless, PPARα/γ dual agonists like muraglitazar, naveglitazar, tesaglitazar, ragaglitazar and aleglitazar have been reported to have undesirable adverse effects, and their developments have been halted at various stages. On the other hand, a recently introduced PPARα/γ dual agonist, saroglitazar is an emerging therapeutic agent of glitazar class approved in India for the management of diabetic dyslipidemia, and its treatment has been reported to be generally safe and well tolerated. Some additional and new compounds, at initial and preclinical stages, have been recently reported to possess PPARα/γ dual agonistic potentials with considerable therapeutic efficacy and reduced adverse profile. This review sheds light on the current status of various PPARα/γ dual agonists for the management of diabetic dyslipidemia.


Facile syntheses of O(2)-[4-(3-pyridyl-4-oxobut-1-yl]thymidine, the major adduct formed by tobacco specific nitrosamine 4-methylnitrosamino-1-(3-pyridyl)-1-butanone (NNK) in vivo, and its site-specifically adducted oligodeoxynucleotides.
O(2)-[4-(3-Pyridyl)-4-oxobut-1-yl]thymidine (O(2)-POB-dThd) is the most persistent adduct detected in the lung and liver of rats treated with tobacco specific nitrosamines: N'-nitrosonornicotine (NNN), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), and its metabolite 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL). It is an important biomarker to assess the human exposure to these carcinogens. The only synthetic method reported for O(2)-POB-dThd requires repeated HPLC purifications and could only be used to prepare an analytical standard due to very low yield (0.4%). We have developed for the first time a regioselective and efficient method for the total synthesis of O(2)-POB-dThd and its site-specifically adducted oligonucleotides. The main step in the synthesis of O(2)-POB-dThd was achieved by a novel method. The treatment of O(2)-5'-anhydrothymidine with the sodium salt of 4-(1,3-dithian-2-yl)-4-(3-pyridyl)butan-1-ol gave exclusively the O(2)-alkylated adduct, which was deprotected in one step to furnish the desired O(2)-POB-dThd in excellent yield. The product was characterized by NMR ((1)H and (13)C), high-resolution MS, and HPLC analysis. This work provided for the first time a reliable method for large scale total synthesis of O(2)-POB-dThd that allowed for solid state site-specifically adducted oligomer synthesis. The O(2)-POB-dThd was converted to its phosphoramidite and subsequently used for the synthesis of oligodeoxynucleotides by standard methods. The oligomers were characterized by MS and HPLC analysis. These oligomers will facilitate the elucidation of the mutagenic potential of the O(2)-POB-dThd adduct, which will provide further insight into the role of tobacco-specific nitrosamines in inducing cancers in smokers.