Is it the Surgeon? A Re-examination of Mid-urethral Sling Complications.
To identify potential technical factors during initial mid-urethral sling (MUS) placement that contribute to subsequent sling revision procedures. A retrospective chart review was performed examining synthetic MUS reoperations at a single institution from 2008-2020. The implanting surgeon's operative note, when available, was critically reviewed with respect to trocar placement, sling location, and tensioning technique. The reoperative report was reviewed for aberrant intraoperative findings relating to the index surgery. A total of 306 women underwent revision of their MUS. Operative reports from the implanting surgeon were available for 276 (90.2%) women. Review of index operative reports revealed 47 unambiguous descriptions of improper technique, including 19 cases of described excessive tensioning and 2 cases of MUS placement despite noted urethral injury during the index case. Indications for reoperation were clinical obstruction (63%), pain (40%), and mesh erosion/exposure (33%). In 186 (67.4%) women, there was an intraoperative finding during the revision that likely contributed to the need for reoperation. Among these 186 women, 110 (59.1%) slings were noted to have been placed too proximally, 78 (41.9%) were over-suspended, and 57 (30.6%) were placed too deep in the periurethral fascia. Review of the index operative note and findings at operative re-exploration of MUS surgeries, often reveals evidence that the initial MUS implantation was technically suboptimal. Such findings suggest that intraoperative surgical technique is a critically important factor contributing to postoperative complications in MUS surgery. This underscores the importance of surgical training and adherence to surgical principles during the placement of a synthetic MUS.


Positive and negative regulatory elements in the upstream region of the rat Cu/Zn-superoxide dismutase gene.
Cu/Zn-superoxide dismutase (SOD1) catalyses the dismutation of superoxide radicals and neutralizes the oxidative effects of various chemicals. Deletion analysis of the upstream region of the rat SOD1 gene revealed that the promoter contains a positive regulatory element (PRE) and a negative regulatory element (NRE), which encompass the regions from -576 to -412 and from -412 to -305 respectively from the site of initiation of transcription. These DNA elements showed enhancer and silencer activities respectively in the natural context and in a heterologous promoter system. Using an electrophoretic-mobility-shift assay and a supershift assay with a specific antibody, the cis-elements of the PRE and NRE were identified as binding sites for transcription factors Elk1 and YY1 (Ying-Yang 1) respectively. Consistent with the presumed roles of the PRE and NRE, Elk1 increased SOD1 gene transcription about 4-5-fold, whereas YY1 exerted a negative effect of about 6-fold. Mutations of the Elk1- and YY1-binding sites led to diminution and elevation respectively of transcriptional activities, both in the natural context and in heterologous promoter systems. These results suggest that the transcription factors Elk1 and YY1, binding in the PRE and NRE respectively, co-ordinate the expression of the SOD1 gene.


The distribution and migration of sodium from a reclaimed upland to a constructed fen peatland in a post-mined oil sands landscape.
Post-mine landscape reclamation of the Athabasca Oil Sands Region requires the use of tailings sand, an abundant mine-waste material that often contains large amounts of sodium (Na+). Due to the mobility of Na+ in groundwater and its effects on vegetation, water quality is a concern when incorporating mine waste materials, especially when attempting to construct groundwater-fed peatlands. This research is the first published account of Na+ redistribution in groundwater from a constructed tailings sand upland to an adjacent constructed fen peat deposit (Nikanotee Fen). A permeable petroleum coke layer underlying the fen, extending partway into the upland, was important in directing flow and Na+ beneath the peat, as designed. Initially, Na+ concentration was highest in the tailings sand (average of 232mgL-1) and lowest in fen peat (96mgL-1). Precipitation-driven recharge to the upland controlled the mass flux of Na from upland to fen, which ranged from 2 to 13tons Na+ per year. The mass flux was highest in the driest summer, in part from dry-period flowpaths that direct groundwater with higher concentrations of Na+ into the coke layer, and in part because of the high evapotranspiration loss from the fen in dry periods, which induces upward water flow. With the estimated flux rates of 336mmyr-1, the Na+ arrival time to the fen surface was estimated to be between 4 and 11years. Over the four-year study, average Na+ concentrations within the fen rooting zone increased from 87 to 200mgL-1, and in the tailings sand decreased to 196mgL-1. The planting of more salt-tolerant vegetation in the fen is recommended, given the potential for Na+ accumulation. This study shows reclamation designs can use layered flow system to control the rate, pattern, and timing of solute interactions with surface soil systems.


AADMA as a marker of endothelial dysfunction in prediabetic women.
to determine the correlation between glycohemoglobin (HbA1c) with level of asymmetric dimethylarginine (ADMA) in serum, between HbA1c and value of brachial artey flow-mediated dilatation (FMD) by ultrasound study, and between ADMA serum and FMD in prediabetes women. the study was done in prediabetes women aged between 30-55 years of age in Cipto Mangunkusumo Hospital Jakarta (RSCM). Prediabetes was based on PERKENI criteria. Subject with fasting blood glucose less than 126 mg/dL and 2-hours blood glucose less than 200 mg/dL met the criteria. Laboratory test of HbA1c and ADMA plasma were performed. To asses brachial FMD, the left brachial artery diameter was measured both at rest and during reactive hyperemia. Increased flow was induced by inflation of pneumatic tourniquet around the forearm to a pressure of 50 mm Hg upper systolic pressure for 5 minutes, followed by release. Measurements of arterial diameter were performed at end diastole at rest and 60 seconds after cuff release. The vessel diameter in scans after reactive hyperemia was expressed as the percentage relative to resting scan (100%). All ultrasound scans were analyzed by a single reader. from 41 prediabetes subjects could be found correlation between ADMA serum with 2-hours post prandial blood glucose (p 0,003 and r 0,457) and HbA1c (p <0,001 and r 0,720). We also found correlation between FMD value with 2-hours post prandial blood glucose (p 0,01 and r -0.487) and HbA1c (p <0,001 and r -0.763). Besides that, there was correlation between ADMA serum with FMD value (p <0,001 and r -0,617). From multivariate analysis, we could determine that HbA1c is the influential factor of ADMA serum and FMD. in prediabetes women there was correlation between HbA1c with ADMA, between HbA1c with FMD and between ADMA serum with FMD.


Acute Effects of a Single-Session of Full-Body Foam Rolling on Heart Rate Variability.
Self-myofascial release has been demonstrated to increase relaxation and improve blood flow, yet it is unknown if it can elicit an acute effect on heart rate variability (HRV). This study aimed to determine if a single-bout of foam rolling could increase parasympathetic activity as measured by HRV. Twenty (20) participants volunteered and their baseline HRV was assessed using a finger sensor while lying supine, and then a second measure was recorded with them sitting upright with feet on the ground. This study utilized a practical HRV collection method designed for use in the field settings, and the data is recorded in arbitrary units (A.U.). Participants then oscillated on a closed-cell cylindrical foam roller using their body weight on the triceps surae, hamstrings, quadriceps, lumbar spine, and pectoral muscles each for 60 sec. Participants first massaged the right limb and then repeated on the matching muscle group on the left before continuing to the next region. Follow-up HRV measurements were recorded using the same procedures. Paired samples t tests assessing pre- and post-foam-rolling HRV measures in supine and seated positions revealed no difference between supine measures (p = .05, d = 0.21), nor the seated measures (p = .27, d = 0.17) among all participants. When sexes were analyzed separately, males showed a significant decrease in HRV in supine positions (p = .03, d = 0.33), but females did not (p = .55, d = 0.09). The single bout of foam rolling on large muscle groups did not increase parasympathetic activity as hypothesized. Males and females responded differently in supine positions, but no difference was present in seated positions across sexes. The task of self-massage may have prevented the anticipated response from the massage. Further research should investigate if passive massage is more effective on HRV response.


Impaired regional left ventricular strain after repair of tetralogy of Fallot.
To test the potential of magnetic resonance imaging (MRI) in early detection of left ventricular (LV) dysfunction in patients with pulmonary regurgitation and normal LV ejection fraction after repair of tetralogy of Fallot. Patients (n = 18) with repaired tetralogy of Fallot and pulmonary regurgitation were prospectively recruited. Healthy volunteers (n = 10) were used as control. Tagging MR images were acquired at the base, mid, and apical LV levels for assessing segmental rotation and circumferential strain. Cine MR images and velocity-encoded MR images were also acquired for assessment of biventricular volumes and biventricular function and pulmonary regurgitant fraction, respectively. Mean values were compared between groups using unpaired Student's t-test. Patients presented with preserved global LV function (LVEF of 59 ± 5%). A significant decrease in LV peak circumferential strain was seen in patients compared with normal volunteers at the basilar (-15.6 ± 4.5% vs. -17.6 ± 4.4%; P < 0.01) and apical (-14.4± 6.1% vs. -17.3± 5.1%, P < 0.01) slices. LV peak rotation was also delayed in patients compared with volunteers at the basilar (6.1 ± 2.6° vs. 4.2 ± 0.6°; P < 0.01) and mid (8.0 ± 1.7° vs. 4.9 ± 1.0°; P < 0.01) slices. MRI can detect early regional LV dysfunction in patients with preserved LVEF after repair of tetralogy of Fallot. MR may be a useful technique for guiding clinical decisions in these patients in order to prevent future global LV deterioration.


Acute diabetes insipidus mediated by vasopressinase after placental abruption.
Postpartum, diabetes insipidus (DI) can be part of Sheehan's syndrome or lymphocytic hypophysitis in combination with anterior pituitary hormone deficiencies. In contrast, acute onset of isolated DI in the postpartum period is unusual. This patient presented at 33 weeks gestation with placental abruption, prompting a cesarean delivery of twins. Immediately after delivery, she developed severe DI. The DI could be controlled with the vasopressinase-resistant 1-deamino-8-D-arginine vasopressin (DDAVP), but not with arginine vasopressin (AVP), and it resolved within a few weeks. The aim of this study was to demonstrate that the postpartum DI in this patient was caused by the release of placental vasopressinase into the maternal bloodstream. Cells were transiently transfected with the AVP receptor 2 (AVPR2) and treated with either AVP or DDAVP in the presence of the patient's serum collected postpartum or 10 weeks after delivery. The response to the different treatments was evaluated by measuring the activity of a cAMP-responsive firefly luciferase reporter construct. The in vitro studies demonstrate that the patient's postpartum serum disrupts activation of the AVPR2 by AVP, but not by the vasopressinase-resistant DDAVP. Placental abruption can rarely be associated with acute postpartum DI caused by release of placental vasopressinase into the bloodstream. This clinical entity must be considered in patients with placental abruption and when evaluating patients presenting with DI after delivery.


INCURVATA2 encodes the catalytic subunit of DNA Polymerase alpha and interacts with genes involved in chromatin-mediated cellular memory in Arabidopsis thaliana.
Cell type-specific gene expression patterns are maintained by the stable inheritance of transcriptional states through mitosis, requiring the action of multiprotein complexes that remodel chromatin structure. Genetic and molecular interactions between chromatin remodeling factors and components of the DNA replication machinery have been identified in Schizosaccharomyces pombe, indicating that some epigenetic marks are replicated simultaneously to DNA with the participation of the DNA replication complexes. This model of epigenetic inheritance might be extended to the plant kingdom, as we report here with the positional cloning and characterization of INCURVATA2 (ICU2), which encodes the putative catalytic subunit of the DNA polymerase alpha of Arabidopsis thaliana. The strong icu2-2 and icu2-3 insertional alleles caused fully penetrant zygotic lethality when homozygous and incompletely penetrant gametophytic lethality, probably because of loss of DNA polymerase activity. The weak icu2-1 allele carried a point mutation and caused early flowering, leaf incurvature, and homeotic transformations of sepals into carpels and of petals into stamens. Further genetic analyses indicated that ICU2 interacts with TERMINAL FLOWER2, the ortholog of HETEROCHROMATIN PROTEIN1 of animals and yeasts, and with the Polycomb group (PcG) gene CURLY LEAF. Another PcG gene, EMBRYONIC FLOWER2, was found to be epistatic to ICU2. Quantitative RT-PCR analyses indicated that a number of regulatory genes were derepressed in the icu2-1 mutant, including genes associated with flowering time, floral meristem, and floral organ identity.


[Dietary modifications, in young Muslims engaged in Ramadan fasting].
The practice of the Islamic precept of Ramadan (R) imposes to those who practice it important physiological and psychological changes due to the water and dietary restrictions they are subjected to, which has a particular impact on the Muslim students' physical and psychological performances, particularly in multicultural Western societies, where there are no schedule adjustments in daily activities. Among the changes in lifestyle, consequently, the dietary changes are stressed, both qualitative and quantitative. Due to the importance of this issue and facing the limited knowledge the Muslim teenagers who live in non-Islamic societies have about the effects of Ramadan, it is suggested as main objective of this study the analysis of changes in nutrients during R in a group of Muslim students of both sexes, in order to solve and prevent some of the problems shown in this period. 55 Muslim youngsters were selected (24 men and 31 women) aged between 15 and 18, who practise R; and the diet was analysed by inventory over 3 days, one week before fasting and in the second week during it. In R there are no relevant changes in caloric intake. The consumption of carbohydrates and proteins is increased, and that of fat is decreased. the changes in R lead to a diet more regulated to nutritional recommendations; and to this seems to have contributed the fact that the participants took their meals at home, where, in spite of the Western cut of the population, it is felt an important influence of North African traditions and habits, due to its border contact with Morocco.


Site-specific dual ink dip pen nanolithography.
The ability to deposit different materials with nanoscale precision at user-specified locations is a very important attribute of dip pen nanolithography (DPN). However, the potential of DPN goes beyond simple deposition since DPN used in conjunction with lateral force microscopy (LFM) allows site-specific investigations of nanoscale properties. In this work, we use two different inks, 16-mercaptohexadecanoic acid (MHA) and 1-octadecanethiol (ODT) to show site-specific dual ink DPN enabled exclusively by our proprietary software. A diamond-dot pattern was created by using a layer-to-layer alignment (LLA) algorithm, which enables a MHA pattern (diamond) to be written concentric with another ODT (central dot) pattern. This simple demonstration of multi-ink DPN is not specific to alkanethiol ink systems, but is also applicable to other multi-material patterning, interaction, and exchange studies.


Regulation of cell adhesion by affinity and conformational unbending of alpha4beta1 integrin.
Rapid activation of integrins in response to chemokine-induced signaling serves as a basis for leukocyte arrest on inflamed endothelium. Current models of integrin activation include increased affinity for ligand, molecular extension, and others. In this study, using real-time fluorescence resonance energy transfer to assess alpha(4)beta(1) integrin conformational unbending and fluorescent ligand binding to assess affinity, we report at least four receptor states with independent regulation of affinity and unbending. Moreover, kinetic analysis of chemokine-induced integrin conformational unbending and ligand-binding affinity revealed conditions under which the affinity change was transient whereas the unbending was sustained. In a VLA-4/VCAM-1-specific myeloid cell adhesion model system, changes in the affinity of the VLA-4-binding pocket were reflected in rapid cell aggregation and disaggregation. However, the initial rate of cell aggregation increased 9-fold upon activation, of which only 2.5-fold was attributable to the increased affinity of the binding pocket. These data show that independent regulation of affinity and conformational unbending represents a novel and fundamental mechanism for regulation of integrin-dependent adhesion in which the increased affinity appears to account primarily for the increasing lifetime of the alpha(4)beta(1) integrin/VCAM-1 bond, whereas the unbending accounts for the increased capture efficiency.


Chromatin-prebound Crm1 recruits Nup98-HoxA9 fusion to induce aberrant expression of Hox cluster genes.
The nucleoporin Nup98 is frequently rearranged to form leukemogenic Nup98-fusion proteins with various partners. However, their function remains largely elusive. Here, we show that Nup98-HoxA9, a fusion between Nup98 and the homeobox transcription factor HoxA9, forms nuclear aggregates that frequently associate with facultative heterochromatin. We demonstrate that stable expression of Nup98-HoxA9 in mouse embryonic stem cells selectively induces the expression of Hox cluster genes. Genome-wide binding site analysis revealed that Nup98-HoxA9 is preferentially targeted and accumulated at Hox cluster regions where the export factor Crm1 is originally prebound. In addition, leptomycin B, an inhibitor of Crm1, disassembled nuclear Nup98-HoxA9 dots, resulting in the loss of chromatin binding of Nup98-HoxA9 and Nup98-HoxA9-mediated activation of Hox genes. Collectively, our results indicate that highly selective targeting of Nup98-fusion proteins to Hox cluster regions via prebound Crm1 induces the formation of higher order chromatin structures that causes aberrant Hox gene regulation.


Medication adherence with fixed-dose versus free-equivalent combination therapies: Systematic review and meta-analysis.
Objective: We conducted a large-scale meta-analysis and subgroup analysis to compare the effect of fixed-dose combination (FDC) therapy with that of free-equivalent combination (FEC) therapy on medication adherence. Methods: Studies published in Web of Science, PubMed, Cochrane Library, ScienceDirect, and Embase up to May 2022 were identified according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. The primary assessed outcomes were the medication possession ratio (MPR) and proportion of days covered (PDC). We investigated the probability of being adherent to the prescribed treatment (MPR or PDC ≥80%) or the average estimate of these two parameters. Studies reporting such results were included in this meta-analysis. The summary measures were reported as the risk ratio (RR) and the weighted mean difference (MD) with 95% of confidence interval (CI) using the random-effects model of DerSimonian and Laird. The quality of the cohort studies was assessed using the Newcastle-Ottawa scale. Results: Of the 1,814 screened studies, 61 met the predefined inclusion criteria. The meta-analysis of the results showed that compared to FEC, FDC significantly improved the medication compliance of patients by 1.29 times (95% CI:1.23-1.35, p < 0.00001). I2 of 99% represent high heterogeneity across studies. The mean difference in medication adherence between FDC and FEC was 0.10 (95% CI: 0.06-0.14, p < 0.00001) with an I2 estimate of 100%. Subgroup analyses were performed for studies that reported adherence outcomes according to disease type, period of evaluation and compliance indicators. A sensitivity analysis was conducted to exclude the results of low-quality studies, as well as studies in which there was ambiguity in the method of calculating the estimator. Conclusion: Analysis of the assessed parameters for the intention-to-treat and subgroup populations suggests that FDC can improve adherence to treatment and its advantages over FEC may increase over time. Further research is needed to better understand how medical conditions affect the impact of reduced pill burden on adherence, particularly in diseases other than cardiovascular disease and type 2 diabetes mellitus.


The UPTAKE study: a cross-sectional survey examining the insights and beliefs of the UK population on COVID-19 vaccine uptake and hesitancy.
A key challenge towards a successful COVID-19 vaccine uptake is vaccine hesitancy. We examine and provide novel insights on the key drivers and barriers towards COVID-19 vaccine uptake. This study involved an anonymous cross-sectional online survey circulated across the UK in September 2020. The survey was designed to include several sections to collect demographic data and responses on (1) extent of agreement regarding various statements about COVID-19 and vaccinations, (2) previous vaccination habits (eg, if they had previously declined vaccination) and (3) interest in participation in vaccine trials. Multinominal logistic models examined demographic factors that may impact vaccine uptake. We used principle component analysis and text mining to explore perception related to vaccine uptake. The survey was circulated through various media, including posts on social media networks (Facebook, Twitter, LinkedIn and Instagram), national radio, news articles, Clinical Research Network website and newsletter, and through 150 West Midlands general practices via a text messaging service. There were a total of 4884 respondents of which 9.44% were black, Asian and minority ethnic (BAME) group. The majority were women (n=3416, 69.9%) and of white ethnicity (n=4127, 84.5%). Regarding respondents, overall, 3873 (79.3%) were interested in taking approved COVID-19 vaccines, while 677 (13.9%) were unsure, and 334 (6.8%) would not take a vaccine. Participants aged over 70 years old (OR=4.63) and the BAME community (OR=5.48) were more likely to take an approved vaccine. Smokers (OR=0.45) and respondents with no known illness (OR=0.70) were less likely to accept approved vaccines. The study identified 16 key reasons for not accepting approved vaccines, the most common (60%) being the possibility of the COVID-19 vaccine having side effects. This study provides an insight into focusing on specific populations to reduce vaccine hesitancy. This proves crucial in managing the COVID-19 pandemic.


[A mechanism of mononucleotide formation under endonuclease hydrolysis].
It has been shown under poly-A hydrolysis by endonuclease A236 in the presence of large amounts of E. coli phosphatase that the formation of mononucleotides requires the presence of terminal 5'-P in the substrate. Simultaneously, it has been found for endonuclease A236 and nuclease of Serratia marcescens that the products of exhaustive hydrolysis carried out in the presence of excess amounts of phosphatase contain an additional nucleoside residue as compared to the ordinary products of exhaustive hydrolysis, the number of phosphate groups being equal in both cases.


Prediction of low bone mass using a combinational approach of cortical and trabecular bone measures from dental panoramic radiographs.
The aim of this study is to extract cortical and trabecular features of the mandible and to develop a novel combinational model of mandibular cortical thickness, trabecular bone area and age in order to predict low bone mineral density or osteoporosis from a dental panoramic radiograph. The study involved 64 south Indian women (age = 52.5 ± 12.7 years) categorised into two groups (normal and low bone mineral density) based on total femur bone mineral density. The dental panoramic radiographs were obtained by a digital scanner, and measurement of total bone mineral density at the right femur was performed by a dual-energy X-ray absorptiometry scanner. The mandibular cortical thickness and panoramic mandibular index were measured bilaterally, and the mean values were considered. The region of interest of 128 × 128 pixels around the mental foramen region was manually cropped and subjected to pre-processing, normalisation and average threshold-based segmentation to determine trabecular bone area. Multiple linear regression analyses of cortical and trabecular measures along with age were performed to develop a combinational model to classify subjects as normal and low bone mineral density. The proposed approach demonstrated strong correlation (r = 0.76; p < 0.01) against the total bone mineral density and resulted in accuracy, sensitivity and positive predictive values of 0.84, 0.92 and 0.85, respectively; the receiver operating characteristic outcomes disclosed that the area under the curve was 0.89.Our results suggest that the proposed combinational model could be useful to diagnose subjects with low bone mineral density.


Critical role of collapsin response mediator protein-associated molecule CRAM for filopodia and growth cone development in neurons.
Collapsin response mediator proteins (CRMPs) have been implicated in signaling of axonal guidance, including semaphorins. We have previously identified a unique member of this gene family, CRMP-associated molecule CRAM (CRMP-5), which is phylogenetically divergent from the other four CRMPs. In this study, we have examined the distribution and function of CRAM in developing neurons. Immunohistochemical analysis showed accumulation of CRAM in the filopodia of growth cones. Experiments using cytochalasin D indicated that filopodial localization of CRAM was independent of filamentous actin. Overexpression of CRAM in neuronal cells significantly promoted filopodial growth and led to the formation of supernumerary growth cones, which acquired resistance to semaphorin-3A stimulation. Finally, knockdown of CRAM by using RNA interference blocked filopodial formation and revealed an aberrant morphology of growth cones. We propose that CRAM regulates filopodial dynamics and growth cone development, thereby restricting the response of growth cone to repulsive guidance cues.


Newly developed hypertension as an early marker of recurrence of adrenocortical carcinoma with high renin expression.
Adrenocortical carcinoma can recur frequently after successful surgery but no adequate markers can detect this recurrence. Here, we present a recurrence of adrenocortical carcinoma with a high renin expression, after successful surgery, where hypertension has developed again. Tumor recurrence was observed via (18)F-fluorodeoxyglucose positron emission tomography and coregistered computed tomography. Based on our findings, follow-up blood pressure assessment may effectively predict the recurrence of adrenocortical carcinoma.


The importance of bleomycin in combination chemotherapy for good-prognosis germ cell carcinoma. Australasian Germ Cell Trial Group.
In an effort to maintain the excellent long-term results achieved with combination chemotherapy for good-prognosis germ cell carcinoma, but to reduce the toxicities encountered, a randomized trial was conducted comparing cisplatin and vinblastine with or without bleomycin. Two hundred eighteen assessable patients with a good prognosis were randomized to receive induction chemotherapy with cisplatin 100 mg/m2 intravenously (IV) day 1 and vinblastine 6 mg/m2 IV days 1 and 2 every 3 weeks (PV) with or without bleomycin 30 mg intramuscularly (IM) weekly (PVB) for a maximum of 12 weeks. Once maximum response was achieved, patients with a complete remission (CR) received two courses of consolidation chemotherapy, while those with residual abnormalities and normal tumor markers underwent surgical resection whenever possible. Toxicities encountered in this study were clearly greater for those patients who received bleomycin, with significantly more leukopenia, thrombocytopenia, anemia, alopecia, and renal and pulmonary toxicities. The proportion of patients who achieved CR and had no evidence of disease (resection of all viable malignancy) was 89% for PV and 94% for PVB (P = .29). After a minimum of 4 years of follow-up, relapses have occurred in 7% of patients who received PV and 5% who received PVB. A total of five patients on each therapy arm were successfully treated with further salvage chemotherapy and surgery. Thus, deaths from progressive malignancy have occurred in 15% of patients on PV and 5% on PVB (P = .02), a rate that was partly offset by the higher proportion of toxic deaths with PVB (P = .06). Despite the toxicities encountered with bleomycin in cisplatin-based combination chemotherapy for these patients, complete deletion of this drug compromises therapeutic efficacy.


The effects of ocean acidification and a carbon dioxide capture and storage leak on the early life stages of the marine mussel Perna perna (Linneaus, 1758) and metal bioavailability.
The study assesses the effects of carbon dioxide capture and storage (CCS) leaks and ocean acidification (OA) on the metal bioavailability and reproduction of the mytilid Perna perna. In laboratory-scale experiments, CCS leakage scenarios (pH 7.0, 6.5, 6.0) and one OA (pH 7.6) scenario were tested using metal-contaminated sediment elutriates and seawater from Santos Bay. The OA treatment did not have an effect on fertilisation, while significant effects were observed in larval-development bioassays where only 16 to 27 % of larva developed normally. In treatments that simulated CO2 leaks, when compared with control, fertilisation success gradually decreased and no larva developed to the D-shaped stage. A fall in pH increased the bioavailability of metals to marine mussels. Larva shell size was significantly affected by both elutriates when compared with seawater; moreover, a significant difference occurred at pH 6.5 between elutriates in the fertilisation bioassay.


The anxiolytic effect of cannabidiol depends on the nature of the trauma when patients with post-traumatic stress disorder recall their trigger event.
We assessed whether administering cannabidiol (CBD) before recalling the traumatic event that triggered their disorder attenuates anxiety in patients with post-traumatic stress disorder (PTSD). As an exploratory pilot analysis, we also investigated whether this effect depends on the nature of the event (sexual vs. nonsexual trauma). Thirty-three patients of both sexes with PTSD were recruited and randomized 1:1 into two groups. One group received oral CBD (300 mg), and the other received a placebo before listening to a digital audio playback of their previously recorded report of the trigger event. Subjective and physiological measurements were taken before and after recall. We analyzed the data in two subsamples: trigger events involving sexual and nonsexual trauma. In the nonsexual trauma group, the differences between measurements before and after recall were significantly smaller with CBD than placebo; this held true for anxiety and cognitive impairment. However, in the sexual trauma group, the differences were non-significant for both measurements. A single dose of CBD (300mg) attenuated the increased anxiety and cognitive impairment induced by recalling a traumatic event in patients with PTSD when the event involved nonsexual trauma.


Dose-effect relations in time-limited psychotherapy for depression.
Criteria for reliable and clinically significant improvement were applied to standard and individually tailored outcome measures data from 212 depressed clients who had been randomly assigned to receive either 8 or 16 sessions of time-limiting psychotherapy. The data were used to address 2 questions: (a) Is the dose-effect curve for psychological symptoms negatively accelerated? and (b) is there a differential rate of response for acute, chronic, and characterological/interpersonal components of depression? The results supported the differential rate of response of different components of depression and suggested qualifications to the acute, chronic, and characterological/interpersonal components and evidence that both supported and qualified previous suggestions that the dose-effect curve is negatively accelerated.


A new family with a case of severe early-onset muscle fatigue and a peculiar maternally inherited painful swelling in chewing muscles associated with homoplasmic m.15992A>T mutation in mitochondrial tRNAPro.
A 16-year-old boy was evaluated for a history of exercise-induced fatigability associated with nausea even after minimal effort, lower limbs muscle hypotrophy, and swelling of the masseter muscles after chewing. Laboratory tests were remarkable for hyperlactatemia and metabolic acidosis after short physical activity. The muscle biopsy showed non-specific mitochondrial alterations and an increase in intrafibral lipids. Biochemical analysis showed reduced activity of the respiratory chain complexes. Mitochondrial DNA sequencing revealed the presence of a homoplasmic variant m.15992A>T in the MT-TP gene, coding for the mt-tRNAPro in the patient, in his mother and in his brother. Pathogenic or likely pathogenic variants in MT-TP gene are rare. They are responsible for different clinical presentation, almost ever involving the muscle tissue. We report the first family with exercise-induced muscle weakness and swelling of the chewing muscles due to m.15992A>T variant in absence of J1c10 haplogroup, confirming its pathogenicity.


The molecular biology of childhood rhabdomyosarcoma.
Rhabdomyosarcomas comprise a biologically heterogeneous group of childhood malignancies that are histologically characterized by varying degrees of differentiation, ranging from uncommitted primitive mesenchymal cells to fetal myotubes. This differentiation pattern is reflected in the behavior of the tumor cells in vitro and can be manipulated by the addition of various biological and chemical agents to the intercellular milieu. Recent studies have documented some of the genetic and phenotypic events that occur under these conditions. In addition, the use of cell lines has facilitated the identification of characteristic karyotypic abnormalities and DNA transcriptional activators that are of potential diagnostic importance. Alterations of tumor-suppressor and multidrug-resistance genes have also been described, and they portend identification of susceptible families and potential therapeutic strategies to deal with this clinically aggressive family of childhood neoplasia. These developments herald an increasing utility of molecular biology in the diagnosis and treatment of pediatric sarcomas and necessitate pathologists' familiarity with this subject.


Glycosylation of sputum mucins is altered in cystic fibrosis patients.
Cystic fibrosis (CF) is characterized by chronic lung infection and inflammation, with periods of acute exacerbation causing severe and irreversible lung tissue damage. We used protein and glycosylation analysis of high-molecular mass proteins in saline-induced sputum from CF adults with and without an acute exacerbation, CF children with stable disease and preserved lung function, and healthy non-CF adult and child controls to identify potential biomarkers of lung condition. While the main high-molecular mass proteins in the sputum from all subjects were the mucins MUC5B and MUC5AC, these appeared degraded in CF adults with an exacerbation. The glycosylation of these mucins also showed reduced sulfation, increased sialylation, and reduced fucosylation in CF adults compared with controls. Despite improvements in pulmonary function after hospitalization, these differences remained. Two CF children showed glycoprotein profiles similar to those of CF adults with exacerbations and also presented with pulmonary flares shortly after sampling, while the remaining CF children had profiles indistinguishable from those of healthy non-CF controls. Sputum mucin glycosylation and degradation are therefore not inherently different in CF, and may also be useful predictive biomarkers of lung condition.


LRIG1‑2 and LMO7 immunoreactivity in vulvar squamous cell carcinoma: Association with prognosis in relation to HPV‑DNA and p16INK4a status.
The present study was conducted to investigate the possible prognostic value of molecular markers LRIG1‑2 and LIM domain 7 protein (LMO7) in vulvar squamous cell carcinoma (VSCC) and their possible correlation to human papilloma virus (HPV)‑ and p16INK4a‑status of the tumors. Patients diagnosed with VSCC at the University Hospital of Umeå, Sweden, during the years 1990‑2013 were selected. Tumor blocks were retrieved from tissue archives and clinical data were collected from the records of patients. HPV‑PCR analysis, HPV genotyping and immunohistochemistry were performed. In total, 112 patients were included. Forty percent of the tumors were HPV‑positive, 27% were p16INK4a‑positive and 23% were positive for both HPV and p16INK4a (considered HPV‑driven). HPV‑positivity and p16INK4a‑positivity were associated with prolonged disease‑free survival (DFS) in Kaplan‑Meier survival analysis. Leucine‑rich repeats and immunoglobulin‑like domains 1 (LRIG1) immunoreactivity was not significantly associated with survival. High leucine‑rich repeats and immunoglobulin‑like domains 2 (LRIG2) immunoreactivity was associated with a prolonged overall survival (OS) (P=0.001). By analyzing HPV‑negative cases only, it was determined that high LRIG2 immunoreactivity was associated with both favorable OS (P=0.008) and DFS (P=0.031). LRIG2 immunoreactivity was also an independent prognostic factor in multivariate analysis of OS (P=0.002, HR=0.41; 95% CI, 0.24‑0.71). High immunoreactivity with LMO7‑1250 antibody was associated with survival benefits in the whole cohort (OS; P=0.011) although DFS was only prolonged in HPV‑negative and not HPV‑driven tumors (P=0.038 and 0.042, respectively). The present study indicated that LRIG2 and LMO7 may be useful prognostic markers in VSCC, particularly for patients without HPV‑driven tumors or with advanced tumors at diagnosis. In contrast to earlier observations regarding other types of squamous cell carcinoma, LRIG1 was not a significant prognostic factor in VSCC.


Freeze-drying of Lactobacillus coryniformis Si3--effects of sucrose concentration, cell density, and freezing rate on cell survival and thermophysical properties.
Freeze-drying is commonly used to stabilize lactic acid bacteria. Many factors have been reported to influence freeze-drying survival, including bacterial species, cell density, lyoprotectant, freezing rate, and other process parameters. Lactobacillus coryniformis Si3 has broad antifungal activity and a potential use as a food and feed biopreservative. This strain is considered more stress sensitive, with a low freeze-drying survival, compared to other commercialized antifungal lactic acid bacterial strains. We used a response surface methodology to evaluate the effects of varying sucrose concentration, cell density and freezing rate on Lb. coryniformis Si3 freeze-drying survival. The water activity of the dry product, as well as selected thermophysical properties of importance for freeze-drying; degree of water crystallization and the glass transition temperature of the maximally freeze concentrated amorphous phase (Tg') were determined. The survival of Lb. coryniformis Si3 varied from less than 6% to over 70% between the different conditions. All the factors studied influenced freeze-drying survival and the most important factor for survival is the freezing rate, with an optimum at 2.8 degrees C/min. We found a co-dependency between freezing rate and formulation ingredients, indicating a complex system and the need to use statistical tools to detect important interactions. The degree of water crystallization decreased and the final water activity increased as a function of sucrose concentration. The degree of water crystallization and Tg' was not affected by the addition of 10(8)-10(10) CFU/ml. At 10(11) CFU/ml, these thermophysical values decreased possibly due to increased amounts of cell-associated unfrozen water.


Numerical and nonnumerical estimation in children with and without mathematical learning disabilities.
There are currently multiple explanations for mathematical learning disabilities (MLD). The present study focused on those assuming that MLD are due to a basic numerical deficit affecting the ability to represent and to manipulate number magnitude ( Butterworth, 1999 , 2005 ; A. J. Wilson & Dehaene, 2007 ) and/or to access that number magnitude representation from numerical symbols ( Rousselle & Noël, 2007 ). The present study provides an original contribution to this issue by testing MLD children (carefully selected on the basis of preserved abilities in other domains) on numerical estimation tasks with contrasting symbolic (Arabic numerals) and nonsymbolic (collection of dots) numbers used as input or output. MLD children performed consistently less accurately than control children on all the estimation tasks. However, MLD children were even weaker when the task involved the mapping between symbolic and nonsymbolic numbers than when the task required a mapping between two nonsymbolic numerical formats. Moreover, in the estimation of nonsymbolic numerosities, MLD children relied more than control children on perceptual cues such as the cumulative area of the dots. Finally, the task requiring a mapping from a nonsymbolic format to a symbolic format was the best predictor of MLD. In order to explain these present results, as well as those reported in the literature, we propose that the impoverished number magnitude representation of MLD children may arise from an initial mapping deficit between number symbols and that magnitude representation.


Churg-Strauss syndrome: high resolution CT and pathologic findings.
The purpose of this study was to evaluate high-resolution CT findings in 7 patients with Churg-Strauss syndrome and to compare the CT with the histopathologic findings. High-resolution CT scans of 7 asthmatic patients (4 women, 3 men, age range, 34-62 years, mean 49 years) with Churg-Strauss syndrome were reviewed by 2 observers. Histologic specimens of lung obtained at surgical (n = 3) or transbronchial (n = 3) biopsy or autopsy (n = 1) were reviewed by an expert lung pathologist. The diagnosis of Churg-Strauss was based on clinical, laboratory, and histologic findings. Parenchymal and airway abnormalities included ground-glass opacities (n = 5), areas of air-space consolidation (n = 4), centrilobular nodules (n = 5), nodules 1-3 cm in diameter (n = 3), interlobular septal thickening (n = 4), bronchial wall thickening (n = 4), and areas of atelectasis (n = 1). Surgical biopsy (n = 3) and autopsy (n = 1) specimens demonstrated airspace disease in 3 patients, interlobular septal thickening in 3 patients, and airway abnormalities in 2 patients. Histologically, the airspace disease included eosinophilic pneumonia (n = 2) and small foci of organizing pneumonia (n = 1). The septal thickening was due to edema combined with numerous (n = 2) or few (n = 1) eosinophils. The airway abnormalities (n = 2) included muscle hypertrophy and large airway wall necrosis (n = 1) and eosinophilic infiltration of the airway walls (n = 1). Transbronchial biopsy (n = 3) demonstrated increased eosinophils. The main high-resolution CT findings of Churg-Strauss syndrome consist of airspace consolidation or ground-glass opacities, septal lines, and bronchial wall thickening. These reflect the presence of eosinophilic infiltration of the airspaces, interstitium, and airways, and interstitial edema.


In vitro hepatotoxicity of the cyanobacterial alkaloid cylindrospermopsin and related synthetic analogues.
Cylindrospermopsin (CY), a sulfate ester of a tricyclic guanidine substituted with a hydroxymethyluracil, is a cyanobacterial toxin of increasing environmental import as it frequently occurs in drinking water reservoirs. As a toxin, CY mainly targets the liver but also involves other organs. In hepatocytes CY inhibits the synthesis of protein and of glutathione, leading to cell death. The total chemical synthesis of CY has recently been reported (Xie et al., 2000, J. Am. Chem. Soc. 22, 5017-5024). The synthesis has provided analogues of CY to study aspects of the relationship between chemical structure and activity that contribute to toxicity. Protein synthesis inhibition was measured in vitro using a rabbit reticulocyte system. Primary cultures of rat hepatocytes were used to determine the biological activity of CY and analogues in intact cells. Protein synthesis and cell glutathione levels were measured. We could distinguish between CY transport and biological activity by comparing the results in vitro to those in intact cells. The role of the sulfate group in CY toxicity was examined by comparing biological effects of CY with that of CY-DIOL (synthetic CY lacking the sulfate group). The sulfate group was found not to play a role in CY activity or in its uptake into cells, since there was no significant difference in biological activity in vitro or in cells between natural CY and CY-DIOL. The orientation of the hydroxyl group at C7 also had no impact on biological activity or transport of CY, since the C7 epimer of CY (EPI-CY) and the corresponding diol (EPI-DIOL) had activity similar to RAC-CY in vitro and in intact cells. AB-MODEL, the analogue lacking an intact C ring, and the methyl and hydroxyl groups of ring A could inhibit protein synthesis (but at concentrations 500-1000-fold higher than natural CY). Other structurally simpler synthetic analogues lacked biological activity.


[Morphometric analysis of the toxic effect of ethanol on the structure of chromatin in interphase nuclei of rat hepatocytes].
A method of automated morphometric analysis of hepatic cell interphase nuclear chromatin (CHIN) structure was elaborated using scanning microspectrophotometer SMP-05 ("Opton", FRG). Liver sections of rats were used to investigate hepatic cell CHIN structure. The animals were administered ethanol intraperitoneally once per day within a month period in doses 4, 6, 8, 10 g/kg (1/3-4/5 LD50). Morphometric indexes indicating dose-dependent changes in hepatic cell CHIN structure are demonstrated. Statistical analysis of morphometric data has led to the conclusion that hepatic cell CHIN structural modification depends on two factors: ethanol damaging effect and hepatic cell regeneration.


The origin of modern metabolic networks inferred from phylogenomic analysis of protein architecture.
Metabolism represents a complex collection of enzymatic reactions and transport processes that convert metabolites into molecules capable of supporting cellular life. Here we explore the origins and evolution of modern metabolism. Using phylogenomic information linked to the structure of metabolic enzymes, we sort out recruitment processes and discover that most enzymatic activities were associated with the nine most ancient and widely distributed protein fold architectures. An analysis of newly discovered functions showed enzymatic diversification occurred early, during the onset of the modern protein world. Most importantly, phylogenetic reconstruction exercises and other evidence suggest strongly that metabolism originated in enzymes with the P-loop hydrolase fold in nucleotide metabolism, probably in pathways linked to the purine metabolic subnetwork. Consequently, the first enzymatic takeover of an ancient biochemistry or prebiotic chemistry was related to the synthesis of nucleotides for the RNA world.


Normative Parameters of Gastrocnemius Muscle Stiffness and Associations with Patient Characteristics and Function.
Quantifying muscle stiffness may aid in the diagnosis and management of individuals with muscle pathology. Therefore, the primary purpose of this study was to establish normative parameters and variance estimates of muscle stiffness in the gastrocnemius muscle in a resting and contracted state. A secondary aim was to identify demographic, anthropometric, medical history factors, and biomechanical factors related to muscle stiffness. Stiffness of the gastrocnemius muscle was measured in both a resting and contracted state in 102 asymptomatic individuals in this cross-sectional study. Differences based on muscle state (resting vs contracted) and sex (female vs male) were assessed using a 2 X 2 analysis of variance (ANOVA). Associations between muscle stiffness and sex, age, BMI, race, exercise frequency, exercise duration, force production, and step length were assessed using correlation analysis. Gastrocnemius muscle stiffness significantly increased from a resting to a contracted state [mean difference: 217.5 (95% CI: 191.3, 243.8), p < 0.001]. In addition, muscles stiffness was 35% greater for males than females in a resting state and 76% greater in a contracted state. Greater muscle stiffness in a relaxed and contracted state was associated with larger plantarflexion force production (r = .26, p < 0.01 and r = .23, p < 0.01 respectively). Identifying normative parameters and variance estimates of muscle stiffness in asymptomatic individuals may help guide diagnosing and managing individuals with aberrant muscle function. 2b Individual Cohort Study. What is known about the subject: Muscle stiffness has been shown to be related to individuals with pathology such as Achilles tendinopathy; however, research is sparse regarding normative values of muscle stiffness. Measuring muscle stiffness may also be a way to potentially predict individuals prone to injury or to monitor the effectiveness of management strategies.What this study adds to existing knowledge: This study establishes defined estimates of muscle stiffness of the gastrocnemius in both a relaxed and contracted state in healthy individuals. Myotonometry measures of muscle stiffness demonstrated an increase in stiffness during contraction that varies by sex. Greater gastrocnemius muscle stiffness was associated with increased plantarflexion force production.


Group X secretory phospholipase A(2) augments angiotensin II-induced inflammatory responses and abdominal aortic aneurysm formation in apoE-deficient mice.
Abdominal aortic aneurysm (AAA) is a complex vascular disease characterized by matrix degradation and inflammation and is a major cause of mortality in older men. Specific interventions that prevent AAA progression remain to be identified. In this study, we tested the hypothesis that Group X secretory phospholipase A(2) (GX sPLA(2)), an enzyme implicated in inflammatory processes, mediates AAA. GX sPLA(2) was detected by immunostaining in human aneurysmal tissue and in angiotensin II (Ang II)-induced AAAs in apolipoprotein E-deficient (apoE(-/-)) mice. GX sPLA(2) mRNA was increased significantly (11-fold) in abdominal aortas of apoE(-/-) mice in response to Ang II infusion. To define the role of GX sPLA(2) in experimental AAAs, apoE(-/-) and apoE(-/-) x GX sPLA(2)(-/-) (GX DKO) mice were infused with Ang II for either 10 (n=7) or 28 (n=24-26) days. Deficiency of GX sPLA(2) significantly reduced the incidence and severity of AAAs, as assessed by ultrasound measurements in vivo of aortic lumens and by computer-assisted morphometric analyses ex vivo of external diameter. Results from gene expression profiling indicated that the expression of specific matrix metalloproteinases and inflammatory mediators was blunted in aortas from GX DKO mice compared to apoE(-/-) mice after 10-day Ang II infusion. Ang II induction of cyclooxygenase-2, interleukin-6, matrix metalloproteinase (MMP)-2, MMP-13 and MMP-14 was reduced significantly in GX DKO mice compared to apoE(-/-) mice. GX sPLA(2) promotes Ang II-induced pathological responses leading to AAA formation.


A survey of green plant tRNA 3'-end processing enzyme tRNase Zs, homologs of the candidate prostate cancer susceptibility protein ELAC2.
tRNase Z removes the 3'-trailer sequences from precursor tRNAs, which is an essential step preceding the addition of the CCA sequence. tRNase Z exists in the short (tRNase ZS) and long (tRNase ZL) forms. Based on the sequence characteristics, they can be divided into two major types: bacterial-type tRNase ZS and eukaryotic-type tRNase ZL, and one minor type, Thermotoga maritima (TM)-type tRNase ZS. The number of tRNase Zs is highly variable, with the largest number being identified experimentally in the flowering plant Arabidopsis thaliana. It is unknown whether multiple tRNase Zs found in A. thaliana is common to the plant kingdom. Also unknown is the extent of sequence and structural conservation among tRNase Zs from the plant kingdom. We report the identification and analysis of candidate tRNase Zs in 27 fully sequenced genomes of green plants, the great majority of which are flowering plants. It appears that green plants contain multiple distinct tRNase Zs predicted to reside in different subcellular compartments. Furthermore, while the bacterial-type tRNase ZSs are present only in basal land plants and green algae, the TM-type tRNase ZSs are widespread in green plants. The protein sequences of the TM-type tRNase ZSs identified in green plants are similar to those of the bacterial-type tRNase ZSs but have distinct features, including the TM-type flexible arm, the variant catalytic HEAT and HST motifs, and a lack of the PxKxRN motif involved in CCA anti-determination (inhibition of tRNase Z activity by CCA), which prevents tRNase Z cleavage of mature tRNAs. Examination of flowering plant chloroplast tRNA genes reveals that many of these genes encode partial CCA sequences. Based on our results and previous studies, we predict that the plant TM-type tRNase ZSs may not recognize the CCA sequence as an anti-determinant. Our findings substantially expand the current repertoire of the TM-type tRNase ZSs and hint at the possibility that these proteins may have been selected for their ability to process chloroplast pre-tRNAs with whole or partial CCA sequences. Our results also support the coevolution of tRNase Zs and tRNA 3'-trailer sequences in plants.


PRS5, the fifth member of the phosphoribosyl pyrophosphate synthetase gene family in Saccharomyces cerevisiae, is essential for cell viability in the absence of either PRS1 or PRS3.
In Saccharomyces cerevisiae, an open reading frame, YOL061w, encodes a polypeptide with sequence similarity to the four known 5-phosphoribosyl-1(alpha)-pyrophosphate synthetase (PRS) genes since it contains a divalent cation binding site and a phosphoribosyl pyrophosphate binding site. We regard YOL061w as the fifth member of the PRS gene family, PRS5. Loss of Prs5p has a significant impact on PRS enzyme activity, causing it to be reduced by 84%. On the other hand, Deltaprs5 strains are not affected in growth or in the size of their nucleotide pools. However, simultaneous deletion of PRS1 and PRS5 or PRS3 and PRS5 rendered the strains inviable, which implies that PRS5 plays an important role in the maintenance of PRS function in S. cerevisiae.


Treatment of multiple myeloma patients with autologous stem cell transplantation - a fresh analysis.
Patients with multiple myeloma (MM) treated with conventional chemotherapy have an average survival of approximately three years. High dose chemotherapy followed by autologous stem cell transplantation (ASCT), first introduced in the mid-1980s, is now considered the standard therapy for almost all patients with multiple myeloma, because it prolongs overall survival and disease free survival. Between November 1997 and October 2006, 122 patients with MM (58 females, 64 males, median age 51.0 years [± 7.98] range: 30-66 years) were transplanted in the Department of Hematooncology and Bone Marrow Transplantation at the Medical University of Lublin: 47 patients were in complete remission or in unconfirmed complete remission, 66 patients were in partial remission, and nine had stable disease. Of these, there were 95 patients with IgG myeloma, 16 with IgA myeloma, one with IgG/IgA, one with IgM myeloma, five with non secretory type, two with solitary tumor and two with LCD myeloma. According to Durie-Salmon, 62 patients had stage III of the disease, 46 had stage II and four had stage I. Most patients (69/122) were transplanted after two or more cycles of chemotherapy, 48 patients were transplanted after one cycle of chemotherapy, one patient after surgery and rtg- -therapy and four patients had not been treated. In mobilisation procedure, the patients received a single infusion of cyclophosphamide (4-6 g/m(2)) or etoposide 1.6 g/m(2) followed by daily administration of G-CSF until the peripheral stem cells harvest. The number of median harvest sessions was 2.0 (± 0.89) (range: 1-5). An average of 7.09 (± 33.28) × 106 CD34(+) cells/kg were collected from each patient (range: 1.8-111.0 × 10⁶/kg). Conditioning regimen consisted of high dose melphalan 60-210 mg/m(2) without TBI. An average of 3.04 (± 11.59) × 10⁶ CD34+ cells/kg were transplanted to each patient. Fatal complications occured in four patients (treatment- -related mortality = 3.2%). In all patients there was regeneration of hematopoiesis. The median number of days for recovery to ANC > 0.5 × 10⁹/l was 13 (± 4.69) (range: 10-38) and platelets recovery to > 50 × 10⁹/l was 25 days (± 11.65) (range: 12-45). Median time of hospitalization was 22 days (± 7.14) (range: 14-50). Patients were evaluated on day 100 after transplantation: 74.9% achieved CR and nCR, 14.3% were in PR, 5.4% had SD and 5.4% had progressed. Median of OS was 45 months (± 30.67). OS at 3-years was 84% and at 7-years 59%. Median PFS was 25 months (± 26.13). PFS at 3-years was 68%, and at 7-years was 43%. At present (November 2009) 52 patients (42%) are still alive. High-dose chemotherapy followed by autologous stem cell transplantation is a valuable, well tolerated method of treatment for patients with MM that allows the achievement of long- -lasting survival.


Factors affecting the implementation of an outcome measurement feedback system in youth mental health settings.
Objective: Measurement feedback systems provide clinicians with regular snapshots of a client's mental health status, which can be used in treatment planning and client feedback. There are numerous barriers to clinicians using outcome measures routinely. This study aimed to investigate factors affecting the use of a measurement feedback system across youth mental health settings. Methods: The participants were 210 clinicians from headspace youth mental health services across Australia. They were surveyed on predictors and use of MyLifeTracker, a routine outcome measure. This was explored through three processes: looking at MyLifeTracker before session, using MyLifeTracker in treatment planning, and providing feedback of MyLifeTracker scores to clients. Results: Clinicians were more likely to look at MyLifeTracker before session, less likely to use it in treatment planning, and least likely to provide MyLifeTracker scores to clients. Each measurement feedback system process had a distinct group of predictors. Perceptions of MyLifeTracker's practicality was the only significant predictor of all three processes. Conclusion: Practically, organisations and supervisors can increase the use of measurement feedback systems through targeted supports.


The Role of Mannose-binding Lectin in Infectious Complications of Pediatric Hemato-Oncologic Diseases.
The complement system is essential for protection against infections in oncologic patients because of the chemotherapy-induced immunosuppression. One of the key elements in the activation of the complement system via the lectin pathway is the appropriate functioning of mannose-binding lectin (MBL) and mannose-binding lectin-associated serine protease 2 (MASP2) complex. The objective of our study was to find an association between polymorphisms resulting in low MBL level and activation of the MBL-MASP2 complex. Also, we aimed at finding a connection between these abnormalities and the frequency and severity of febrile neutropenic episodes in children suffering from hemato-oncologic diseases. Ninety-seven patients had been enrolled and followed from the beginning of the therapy for 8 months, and several characteristics of febrile neutropenic episodes were recorded. Genotypes of 4 MBL2 polymorphisms (-221C/G, R52C, G54D, G57E) were determined by real-time polymerase chain reaction. Activation of the MBL-MASP2 complex was evaluated by enzyme-linked immunosorbent assay at the time of diagnosis and during an infection. The number of febrile neutropenic episodes was lower, and the time until the first episode was longer in patients with normal MBL level than in patients with low MBL level coding genotypes. The MBL-MASP2 complex activation level correlated with the MBL genotype and decreased significantly during infections in patients with low MBL level. Our results suggest that infections after immunosuppression therapy in children suffering from hemato-oncologic diseases are associated with the MBL2 genotype. Our results may contribute to the estimation of risk for infections in the future, which may modify therapeutic options for individuals.


High-order soliton evolution and pulse breaking-recovery in stretched ultrafast fiber lasers.
We present a new pulse regime in a stretched ultrafast fiber laser based on numerical simulations. The pulse breaking due to high-order soliton evolution in the passive fiber is recovered to a smooth pulse in the gain fiber with normal dispersion. The new pulse regime formed by the two nonlinear processes makes the ultrafast fiber laser generate ultra-broadband, ultrashort duration, high energy and large breathing ratio pulses. Our work gives insights into the nonlinear dynamics in fiber lasers and has potential for a better design of the stretched fiber lasers.


Assessment of Thermosonication as Postharvest Treatment Applied on Whole Tomato Fruits: Optimization and Validation.
Tomatoes are a popular and rich fruit due to their nutritional and bioactive composition as vitamins, antioxidants, and phenolics contributing to the promotion of consumer health. For this reason, emerging postharvest technologies need to be evaluated to achieve the maintenance of sensorial and quality-related characteristics, like color and texture, while aiding to fruit decontamination. Optimization of thermosonication as postharvest treatments on whole, mature-green tomatoes (cv. "Zinac") to improve quality (color, texture, total phenolic content, and weight loss) was performed by response surface methodology. Temperature (32-48 °C), treatment time (13-47 min), and storage period at 10 °C (1-15 days) at constant ultrasound frequency (45 kHz; 80% power level), were the independent variables. In general, thermosonication delayed tomato color changes while achieving total phenolic content increase and good overall quality. Three optimal thermosonication conditions were selected and validated (32 °C-13 min, 35 °C-20 min and 40 °C-30 min). The most suitable thermosonication condition that promoted a longer storage while keeping a high-quality standard was at 40 °C during 30 min. This study demonstrated that thermosonication provides an effective alternative methodology to guarantee tomato quality without significant change during the expected postharvest period.


Care Facilitation Advances Movement Along the Hepatitis C Care Continuum for Persons With Human Immunodeficiency Virus, Hepatitis C, and Substance Use: A Randomized Clinical Trial (CTN-0064).
Direct-acting antivirals can cure hepatitis C virus (HCV). Persons with HCV/HIV and living with substance use are disadvantaged in benefiting from advances in HCV treatment. In this randomized controlled trial, participants with HCV/HIV were randomized between February 2016 and January 2017 to either care facilitation or control. Twelve-month follow-up assessments were completed in January 2018.Care facilitation group participants received motivation and strengths-based case management addressing retrieval of HCV viral load results, engagement in HCV/HIV care, and medication adherence. Control group participants received referral to HCV evaluation and an offer of assistance in making care appointments. Primary outcome was number of steps achieved along a series of 8 clinical steps (eg, receiving HCV results, initiating treatment, sustained virologic response [SVR]) of the HCV/HIV care continuum over 12 months postrandomization. Three hundred eighty-one individuals were screened and 113 randomized. Median age was 51 years; 58.4% of participants were male and 72.6% were Black/African American. Median HIV-1 viral load was 27 209 copies/mL, with 69% having a detectable viral load. Mean number of steps completed was statistically significantly higher in the intervention group vs controls (2.44 vs 1.68 steps; χ 2 [1] = 7.36, P = .0067). Men in the intervention group completed a statistically significantly higher number of steps than controls. Eleven participants achieved SVR with no difference by treatment group. The care facilitation intervention increased progress along the HCV/HIV care continuum, as observed for men and not women. Study findings also highlight continued challenges to achieve individual-patient SVR and population-level HCV elimination. NCT02641158.


E-PASS Scoring System May Be Useful for Prediction of Postoperative Complications in Super Elderly Colorectal Cancer Surgery Patients.
Japan is facing an aging society. Elderly individuals are generally more prone to comorbidities and have weaker immune defenses, with ominous prognostic implications if postoperative complications arise. The aim of this study was to explore scoring systems for predicting postoperative morbidity risk in super elderly patients (≥85 years old) after colorectal surgery for cancer. A population of elderly patients (n = 145) surgically treated for primary colorectal cancer within our department between April 2007 and December 2018 was examined retrospectively, assessing the capacities of various indices, such as Estimation of Physiologic Ability and Surgical Stress (E-PASS), neutrophil/lymphocyte ratio (NLR), platelet/lymphocyte ratio (PLR), prognostic nutritional index (PNI), and modified Glasgow Prognostic Score (mGPS), to predict postoperative complications. NLR, PLR, and mGPS did not differ significantly in the presence or absence of complications, whereas PNI tended to be lower if complications developed. The E-PASS system showed no group-wise differences in preoperative risk score (PRS), but the surgical stress score (SSS) and the comprehensive risk score (CRS; a composite of PRS and SSS) was significantly higher in patients with complications. Based on the cutoff value calculated from the Receiver operating curve (ROC) for the E-PASS CRS (-0.0580), patients were stratified into low-scoring and high-scoring (HSG) groups. Although not significantly different, the overall survival in the HSG tended to be lower by comparison. The E-PASS scoring system may be a useful predictor of postoperative complications in super elderly patients requiring colorectal cancer surgery.


Current epidemiology and growing resistance of gram-negative pathogens.
In the 1980s, gram-negative pathogens appeared to have been beaten by oxyimino-cephalosporins, carbapenems, and fluoroquinolones. Yet these pathogens have fought back, aided by their membrane organization, which promotes the exclusion and efflux of antibiotics, and by a remarkable propensity to recruit, transfer, and modify the expression of resistance genes, including those for extended-spectrum β-lactamases (ESBLs), carbapenemases, aminoglycoside-blocking 16S rRNA methylases, and even a quinolone-modifying variant of an aminoglycoside-modifying enzyme. Gram-negative isolates--both fermenters and non-fermenters--susceptible only to colistin and, more variably, fosfomycin and tigecycline, are encountered with increasing frequency, including in Korea. Some ESBLs and carbapenemases have become associated with strains that have great epidemic potential, spreading across countries and continents; examples include Escherichia coli sequence type (ST)131 with CTX-M-15 ESBL and Klebsiella pneumoniae ST258 with KPC carbapenemases. Both of these high-risk lineages have reached Korea. In other cases, notably New Delhi Metallo carbapenemase, the relevant gene is carried by promiscuous plasmids that readily transfer among strains and species. Unless antibiotic stewardship is reinforced, microbiological diagnosis accelerated, and antibiotic development reinvigorated, there is a real prospect that the antibiotic revolution of the 20th century will crumble.


Posttranslational modification of class III beta-tubulin.
The charge heterogeneity of class III beta-tubulin (beta III) during neural development was analyzed by high-resolution isoelectric focusing/two-dimensional polyacrylamide gel electrophoresis in combination with site-specific proteolytic digestion and immunological detection. The number of beta III isoforms (charge variants) gradually increases from one in embryonic brain to seven in adult brain. All of the charge heterogeneity is due to posttranslationally modified sites located within the extreme C-terminal region of the beta III polypeptide. One beta III isoform is present in testis, the only other tissue in which this isotype is expressed. The testis beta III isoform cofocuses with the earliest-appearing embryonic brain beta III charge variant. Our results indicate that the posttranslational modifications of beta III are developmentally regulated, occur at more than one site, and are neuron-specific. The location of these modifications within the extreme C-terminal domain suggests that their function is to modulate the interaction of tubulin with microtubule-associated proteins.


Direct role of HLA-B5 in influencing susceptibility to Takayasu aortoarteritis.
To investigate genetic factors involved in the pathogenesis of Takayasu arteritis (TA), North Indian patients belonging to the states of Punjab, Haryana, Uttar Pradesh and Delhi were examined for HLA-class I and class II antigens and the data compared with healthy controls from the same ethnic group. Additionally, DNA typing was performed using polymerase chain reaction/sequence specific oligonucleotide probe (PCR-SSOP) technique to evaluate the distribution of molecular alleles in the healthy Indian population as compared to orientals and Western caucasoids. The frequency of HLA-B5 was significantly increased in patients as compared to controls (chi 2 = 32.5, corrected P value, Pc = 3 x 10(-6), relative risk = 4.3). Serological splitting of B5 into B51 and B52 did not reveal an association with any of the two subtypes. Weak association was also noticed with DR8 in the patient group (chi 2 = 8.2, Pc = 0.05). Distribution of the molecular subtypes of various HLA-B and DR alleles indicated that although the Indian population is essentially caucasoid, it comprises of an admixture of both caucasoid as well as oriental alleles/haplotypes. The observations clearly suggest that (a) HLA-linked genes are involved in the development of Takayasu arteritis, and (b) rather than a subtype of B5, the whole molecule or its closely linked gene(s) influence susceptibility to TA.


Endothelial Stromal PD-L1 (Programmed Death Ligand 1) Modulates CD8+ T-Cell Infiltration After Heart Transplantation.
The role of checkpoint axes in transplantation has been partially addressed in animal models but not in humans. Occurrence of fulminant myocarditis with allorejection-like immunologic features in patients under anti-PD1 (programmed death cell protein 1) treatment suggests a key role of the PD1/PD-L1 (programmed death ligand 1) axis in cardiac immune homeostasis. We cross-sectionally studied 23 heart transplant patients undergoing surveillance endomyocardial biopsy. Endomyocardial tissue and peripheral blood mononuclear cells were analyzed by flow cytometry. Multivariate logistic regression analyses including demographic, clinical, and hemodynamic parameters were performed. Murine models were used to evaluate the impact of PD-L1 endothelial graft expression in allorejection. We found that myeloid cells dominate the composition of the graft leukocyte compartment in most patients, with variable T-cell frequencies. The CD (cluster of differentiation) 4:CD8 T-cell ratios were between 0 and 1.5. The proportion of PD-L1 expressing cells in graft endothelial cells, fibroblasts, and myeloid leukocytes ranged from negligible up to 60%. We found a significant inverse logarithmic correlation between the proportion of PD-L1+HLA (human leukocyte antigen)-DR+ endothelial cells and CD8+ T cells (slope, -18.3 [95% CI, -35.3 to -1.3]; P=0.030). PD-L1 expression and leukocyte patterns were independent of demographic, clinical, and hemodynamic parameters. We confirmed the importance of endothelial PD-L1 expression in a murine allogeneic heart transplantation model, in which Tie2Crepdl1fl/fl grafts lacking PD-L1 in endothelial cells were rejected significantly faster than controls. Loss of graft endothelial PD-L1 expression may play a role in regulating CD8+ T-cell infiltration in human heart transplantation. Murine model results suggest that loss of graft endothelial PD-L1 may facilitate alloresponses and rejection.


Simulating the mechanics behind sub-optimal mobility and the associated economic losses in dairy production.
Hoof disorders and sub-optimal mobility (SOM) are economically important health issues in dairy farming. Although the dynamics of hoof disorders have an important effect on cow mobility, they have not been considered in previous simulation models that estimate the economic loss of SOM. Furthermore, these models do not consider the varying severities of SOM. The objective of this study was to develop a novel bio-economic simulation model to simulate the dynamics of 8 hoof disorders: digital dermatitis (DD), interdigital hyperplasia (HYP), interdigital dermatitis/heel-horn erosion (IDHE), interdigital phlegmon (IP), overgrown hoof (OH), sole haemorrhage (SH), sole ulcer (SU) and white-line disease (WLD), their role in SOM, and estimate the economic loss of SOM in a herd of 125 dairy cows. A Reed-Frost model was used for DD and a Greenwood model for the other 7 hoof disorders. Economic analysis was conducted per mobility score according to a 5-point mobility scoring method (1 = perfect mobility; 5 = severely impaired mobility) by comparing a scenario with SOM and one without SOM. Parameters used in the model were based on literature and expert opinion and deemed credible during model validation rounds. Results showed that the mean cumulative incidence for maximum mobility scores 2-5 SOM episodes were respectively 34, 16, 7 and <1 episodes per 100 cows per pasture period and 39, 19, 8, <1 episodes per 100 cows per housing period. The mean total annual economic loss due to SOM resulting from the hoof disorders under study was €15,342: €122 per cow per year. The economic analysis uncovered direct economic losses that could be directly linked to SOM episodes and indirect economic losses that could not be directly linked to SOM episodes but arose due to the presence of SOM. The mean total annual direct economic loss for maximum mobility score 2-5 SOM episodes was €1129, €3098, €4354 and €480, respectively. The mean total annual indirect economic loss varied considerably between the 5th and 95th percentiles: €-6174 and €19,499, and had a mean of €6281. This loss was composed of additional indirect culling due to SOM (∼65%) and changes in the overall herd milk production (∼35%) because of additional younger replacement heifers entering the herd due to increased culling rates. The bio-economic model presented novel results with respect to indirect economic losses arising due to SOM. The results can be used to stimulate farmer awareness and promote better SOM management.


Effect of necrosectomy and vacuum-assisted closure (VAC) on mitochondrial function and oxidative stress markers in severe acute pancreatitis.
Severe acute pancreatitis (SAP) is associated with high morbidity and mortality. To evaluate whether necrosectomy, alone or combined with vacuum-assisted closure (VAC), has any additional beneficial effects on mitochondrial function and/or oxidative stress markers in SAP. Patients with SAP, APACHE II score > 8, and inadequate response to management in an intensive care unit were included in a prospective observational study. Sixteen underwent necrosectomy and 24 underwent necrosectomy plus VAC every 48 h. Patients were then categorized as survivors or deceased. Submitochondrial membrane fluidity of platelets and F0F1-ATPase hydrolysis were measured to represent mitochondrial function. Oxidative/nitrosative stress was measured using lipoperoxides (LPOs), nitric oxide (NO), erythrocyte membrane fluidity, and total antioxidant capacity (TAC). Membrane fluidity in submitochondrial particles of platelets remained significantly increased throughout the study, and then eventually rised in deceased patients managed with necrosectomy + VAC vs. survivors (p < 0.041). Hydrolysis was significantly increased from baseline to endpoint in all patients, predominating in those who died after management with necrosectomy (p < 0.03). LPO increased in all patients, and necrosectomy was more efficient for the eventual decrease in survivors (p < 0.039). NO was found to be increased for the baseline-endpoint result among both survivors and deceased patients with both management options. Erythrocyte membrane fluidity was increased in survivors managed with necrosectomy + VAC, and eventually returned to normal (p < 0.045). TAC was found to be consumed in all patients for the duration of the study. Mitochondrial dysfunction and oxidative/ nitrosative stress with significant systemic antioxidant consumption were found. Necrosectomy was more efficient and better cleared LPOs. Necrosectomy + VAC improved erythrocyte membrane fluidity and increased survival.


The cyanotrophic bacterium Pseudomonas pseudoalcaligenes CECT5344 responds to cyanide by defence mechanisms against iron deprivation, oxidative damage and nitrogen stress.
Two-dimensional (2-D) electrophoresis approach has been used to test protein expression changes in response to cyanide in the alkaliphilic bacterium Pseudomonas pseudoalcaligenes CECT5344. This is a cyanide-assimilating strain which also grows in media containing cyanide-enriched effluent from the jewellery industry. The bacterium efficiently uses this residue as the sole nitrogen source for aerobic growth under alkaline pH with negligible nitrogen losses as HCN. Cell-free extracts isolated from P. pseudoalcaligenes grown with a jewellery residue, free cyanide or ammonium chloride as nitrogen source were subjected to 2-D electrophoresis and the spot patterns were examined to determine differential protein expression. Electrophoretic plates exhibiting an average of 1000 spots showed significant differences in the expression of about 44 proteins depending on the nitrogen source. Some of these protein spots were analysed by Matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS). Characterization of five of these proteins reveals that cyanide shock induces proteins related to iron acquisition, regulation of nitrogen assimilation pathways and oxidative stress repairing and protection.


Management of diabetes resistant to subcutaneous insulin with intravenous insulin via an implanted infusion pump.
Diabetes resistant to conventional subcutaneous insulin injection is a rare complication of insulin-dependent diabetes which poses a major management problem. We report three cases treated for a total of over seven patient years with fully implanted insulin infusion devices. Technical difficulties with the devices and their operation have been substantial but the patients are much improved and hospitalisation has been dramatically reduced. We suggest that implanted insulin pumps are a real treatment option for patients with this unusual syndrome.


Effects of boron supplementation on bone mineral density and dietary, blood, and urinary calcium, phosphorus, magnesium, and boron in female athletes.
The effects of boron supplementation on blood and urinary minerals were studied in female college students--17 athletes and 11 sedentary controls--over a one-year period. The athletes had lower percent body fat and higher aerobic capacities than sedentary controls. Athletic subjects consumed more boron in their normal diets than sedentary subjects; all other dietary measures were similar between the two groups. The athletes showed a slight increase in bone mineral density, whereas the sedentary group showed a slight decrease. Serum phosphorus concentrations were lower in boron-supplemented subjects than in subjects receiving placebos, and were lower at the end of the study period than during baseline analysis. Activity depressed changes in serum phosphorus in boron-supplemented subjects. Serum magnesium concentrations were greatest in the sedentary controls whose diets were supplemented with boron, and increased with time in all subjects. A group x supplement interaction was observed with serum magnesium; exercise in boron-supplemented subjects lowered serum magnesium. In all subjects, calcium excretion increased over time; in boron-supplemented subjects, boron excretion increased over time. In all subjects, boron supplementation affected serum phosphorus and magnesium, and the excretion of urinary boron.


Relationship Between Accelerometer-Based Physical Activity, Sedentary Behavior, and Mental Health in Young Finnish Men.
Healthy lifestyle behaviors including physical activity (PA) have been recognized to contribute positively to mental health. Most of the evidence on relationship between PA and mental health relies on self-reported PA results. Device-based measures on PA or sedentary behavior (SB) are less frequently used in mental health research. The present study aimed at examining the relationship between mental health and PA/SB measured by accelerometers in young Finnish men. The sample consisted of 409 men (mean age 28 ± 7 years), who participated in the military refresher training in Finland. Self-rated mental health was measured with Mental Health Inventory (MHI-5) and short Warwick-Edinburgh Mental Well-being Scale (SWEMWBS) measuring mental health both from the perspective of mental health problems and mental well-being. PA was measured with accelerometer from the perspective of light, moderate, vigorous, and total activity, as well as SB. Linear regression models and compositional analysis were applied. Age, education, marital status, employment status, BMI, alcohol use and smoking were used as covariates. Evidence on relationship between total PA (standardized regression coefficient 0.340; 95% CI 0.022-0.657, p = 0.036) and SB (standardized regression coefficient -0.340; 95% CI -0.657 to -0.022, p = 0.036) with symptoms of mental health problems was found after adjusting for age, education, marital and employment status. The relationship was marginally significant (p = 0.056) after adjusting also for BMI, alcohol use and smoking. No evidence on relationship between PA or SB and mental well-being was found, neither in standard linear regression analysis nor in compositional approach. In our sample of young adult men, PA seemed to have a stronger relationship with symptoms of mental health problems rather than with mental well-being. The findings lead to a conclusion that all PA per se may not be independently associated with mental well-being in young adult males and raise the question whether the domain of PA and its context play a critical role in these relationships.


Prevalence of Parkinson Disease in Italy: a systematic review and meta-analysis.
Parkinson's disease (PD) is a common disease of unknown etiology. Even though accurate information on the epidemiology of PD is critical for defining appropriate health policies, epidemiological data on Parkinson's disease (PD) in Italy are often defined as scant or conflicting. Our study attempted to provide an overview on the prevalence of (PD) by means of a systematic review and metanalysis of existing data. Material and methods. We searched into two different databases (PubMed and EMBASE), focusing on studies reporting the prevalence of PD in Italy. Data were extracted using a standardized assessment form, and results of such analyses were systematically reported, summarized and compared. Results. A total of 16 studies were eventually included in the analyses, with a prevalence rate of 193.7/100,000. Available reports were heterogeneous both in design and in eventual figures, and also prevalence estimates were affected by substantial heterogeneity. Interestingly, prevalence rates ranged from 37.8/100,000 inhabitants in subjects aged 0 to 64 years, to 578.7 in age group 65 to 75 years, and 1235.7 in age group 75 years or older. PD was significantly associated with male sex, but only in older age groups (i.e. Odds Ratio, OR 1.37 95%CI 1.22-1.53, and OR 1.31, 95%CI 1.21-1.42 for age groups 65-74 years and 75 years or more, respectively). Discussion and conclusion. While the observed variations in prevalence rates may result from environmental or genetic factors, differences in methodologies for case ascertainment and diagnostic criteria may have significantly affected our estimates. As a consequence, the comparability of existing studies is limited.


HOXB13 induces growth suppression of prostate cancer cells as a repressor of hormone-activated androgen receptor signaling.
Androgen receptor (AR) signals play a decisive role in regulating the growth and differentiation of both normal and cancerous prostate cells by triggering the regulation of target genes, in a process in which AR cofactors have critical functions. Because of the highly prostate-specific expression pattern of HOXB13, we studied the role of this homeodomain protein in prostate cells. Expression of HOXB13 was limited to AR-expressing prostate cells. Reporter transcription assay demonstrated that HOXB13 significantly suppressed hormone-mediated AR activity in a dose-responsive manner, and suppression was specific to AR with which HOXB13 physically interacts. Overexpression of HOXB13 further down-regulated the androgen-stimulated expression of prostate-specific antigen, and suppression of endogenous HOXB13 stimulated transactivation of AR. Functionally, HOXB13 suppressed growth of LNCaP prostate cancer cells, which could be counteracted by additional hormone-activated AR. On the other hand, the growth-suppressive function of HOXB13 in AR-negative CV-1 cells was not affected by AR. These results suggest that HOXB13 functions as an AR repressor to modulate the complex AR signaling and subsequent growth regulation of prostate cancer cells. In addition to the loss of HOXB13 expression, maintaining AR may be an important step for prostate cancer cells to tolerate the suppressor function of HOXB13. Altogether, our data present a novel mechanism for the HOXB13-mediated repression of AR signaling, which can be interpreted to a growth-suppressive event.


Evaluating the Accuracy of Two Microleakage Assessment Methods for Mineral Trioxide Aggregate and Calcium-enriched Mixture Cement.
Multiple methods for evaluating microleakage have been introduced over the years, but there has been no agreement as to which technique will give more accurate results. The aim of this study was to compare the accuracy and results of fluid filtration and marginal adaptation methods for mineral trioxide aggregate (MTA) and calcium-enriched mixture (CEM) cement apical plugs. A total of 250 single-rooted human teeth were collected. The teeth were decoronated, the root canals were prepared and open apex condition was stimulated by passing #1 to 4 Peeso Reamer drills from apical foramen. Five teeth were selected as the positive and negative controls and the rest of the samples were randomly allocated to two groups of MTA and CEM cement plugs. In each group, apical plug was placed into the canal. After the apical plugs were completely set, microleakage and marginal adaptation of the samples were evaluated using fluid filtration method and scanning electron microscopy (SEM), respectively. The obtained results were analyzed by independent-samples t test. Gap between plug and dentin walls and air bubbles displacement was higher in MTA group compared to the CEM cement group, though this difference between MTA group and CEM cement group was not statistically significant. Conclusion: According to the obtained results, it seems that there is a direct relationship between the two methods of microleakage assessment.


DNA assisted self-assembly of PAMAM dendrimers.
We report DNA assisted self-assembly of polyamidoamine (PAMAM) dendrimers using all atom Molecular Dynamics (MD) simulations and present a molecular level picture of a DNA-linked PAMAM dendrimer nanocluster, which was first experimentally reported by Choi et al. (Nano Lett., 2004, 4, 391-397). We have used single stranded DNA (ssDNA) to direct the self-assembly process. To explore the effect of pH on this mechanism, we have used both the protonated (low pH) and nonprotonated (high pH) dendrimers. In all cases studied here, we observe that the DNA strand on one dendrimer unit drives self-assembly as it binds to the complementary DNA strand present on the other dendrimer unit, leading to the formation of a DNA-linked dendrimer dimeric complex. However, this binding process strongly depends on the charge of the dendrimer and length of the ssDNA. We observe that the complex with a nonprotonated dendrimer can maintain a DNA length dependent inter-dendrimer distance. In contrast, for complexes with a protonated dendrimer, the inter-dendrimer distance is independent of the DNA length. We attribute this observation to the electrostatic complexation of a negatively charged DNA strand with the positively charged protonated dendrimer.


Ion Mobility Spectrometry and Tandem Mass Spectrometry Analysis of Estradiol Glucuronide Isomers.
Estradiol is an estrogenic steroid that can undergo glucuronidation at two different sites, which results in two estradiol glucuronide regioisomers. These isomers can be produced by different enzymes and can have different biological activities before being eliminated from the body. Although there have been previous methods that can distinguish the two isomers, these methods often have long acquisition times or high cost per analysis. In this study, traveling wave ion mobility spectrometry (TWIMS) coupled with mass spectrometry (MS) was employed to separate estradiol glucuronides using alkali metal adduction in positive ion mode, where the sodiated dimer adduct provided adequate separation both in single-component standards and in two-component mixtures. Additionally, in negative ion mode, tandem mass spectrometry (MS/MS) was used to quantitatively determine the relative composition of the two isomers. This was possible due to differences in the energetic requirements for loss of the glucuronic acid, which was characterized by energy-resolved collision-induced dissociation (CID). This work demonstrated that the intensity of the glucuronic acid neutral loss product as compared with the intensity of the intact precursor ion can be used to determine the percentage of each isomer present in a mixture. Overall, TWIMS successfully separated estradiol glucuronide isomers in positive ion mode and MS/MS via CID enables relative quantitation of each isomer in negative ion mode.


Effects of time and duration of exposure to 12% O2 and prior food deprivation on hypoxic hypophagia of rats.
A low cost, gas-dilution type, high-altitude simulator was developed and found to be a useful model for studying the effect of acute hypoxic exposure on food consumption in the rat. A distinct advantage of this system is that it provides access to the animals without disturbance to the gaseous atmosphere. The onset of hypoxic hypophagia was shown to occur within the first 3 h following abrupt ascent to a simulated altitude of 4419.6 m (14,500 ft). The magnitude of the anorexic effect was uniform throughout the first day of exposure to 12% O2; likewise, food consumption rapidly recovered following return to a sea level atmosphere. It was concluded that the hypophagic effect of hypoxia is not absolute and that the severity of the hypophagia can be reduced by prior food deprivation. However, the degree of attenuation of the hypophagic response was not directly related to the length of the deprivation period. Food deprivation may sufficiently enhance metabolic-induced food demand signals to partially override the inhibitory effect of hypoxia on feeding behavior. Alternatively, metabolic perturbations associated with the food-deprived state may, in some undefined manner, minimize the severity of acute hypoxic stress.


Some problems with the anti-prohibitionist position on legalization of drugs.
The pro-legal position has mounted cogent arguments to support conclusions that the existing prohibition policy has failed and that legalization, while not a solution to the drug use problem, will effectively eliminate drug related crime. The premise of this article is that a legalization policy is wrong in that the basic assumptions underlying the anti-prohibitionist position are flawed. Prohibition has operated in a social vacuum. It has been an isolated effort substituting for an integrated and well coordinated approach which includes prevention, treatment as well as enforcement and supported by an educated public resolve against illicit drugs.


Mortality from skin melanoma in Italy and Friuli-Venezia Giulia region, 1970-1989.
Substantial upward trends for skin cancer mortality have been observed in most European countries in the last two decades. The distinction, however, between cutaneous malignant melanoma (CMM) and non-melanomatous skin cancers on the basis of death certification is unreliable. In order to assess the trends of CMM, analysis of the specific code for CMM (International Classification of Disease VIII and IX:172) was coupled with analysis of total skin cancer mortality rates, standardized on the 1981 Italian Census population, at all ages and in selected age groups (15-44, 45-64 and 65-84 years). The analyses were carried out for the whole of Italy and for Friuli-Venezia Giulia (FVG), a region in North-East Italy with a high proportion of fair-complexioned individuals, elevated frequency of sunbathing, and especially good diagnostic standards. In 1985-89 skin cancer mortality rates in men at all ages, as compared to 1970-74, were 33% higher in Italy and 46% higher in FVG. In women, there was a rise of 22% in Italy and 94% in FVG. More marked upward trends were observed in young and middle aged adults, especially among women in FVG (over 300% increase). In absolute terms, approximately 3000 more Italians died from skin cancers in 1985-89 as compared to 1970-74, about half of them prematurely (i.e., < 65 years). CMM was mentioned in death certifications for skin cancer increasingly often in all age groups, particularly in FVG. The analysis of mortality trends from skin cancer is consistent with a substantial rise of CMM-related deaths. The sensitivity of death certification with respect to CMM diagnosis is also increasing, especially in FVG. A public health strategy with the aim of reducing intermittent intense exposure to ultraviolet light, especially in children, is urgent in Italy as CMM represents one of the few causes of premature death with unfavorable trends.


Influence of propranolol coadministration or cigarette smoking on the kinetics of desmethyldiazepam following intravenous clorazepate.
The influence of propranolol coadministration or of cigarette smoking on the kinetics of desmethyldiazepam following a single 20-mg intravenous dose of clorazepate dipotassium was evaluated in healthy volunteers. In Study One, intravenous clorazepate was given once in the control condition, and again during coadministration of propranolol, 80 mg twice daily. Compliance with the prescribed propranolol regimen was verified by measurement of serum propranolol concentrations (mean, 37 ng/ml). In control vs propranolol treatment conditions, there was no significant difference in desmethyldiazepam volume of distribution (1.27 vs 1.23 liters/kg) or in free fraction in serum (1.83 vs 1.80% unbound). There was a small although statistically significant prolongation of desmethyldiazepam half-life (55 vs 61 h, P less than 0.05) and reduction in clearance (0.281 vs 0.247 ml/min/kg, P less than 0.02) attributable to propranolol. In Study Two, desmethyldiazepam kinetics were compared in eight cigarette smokers (mean, 19 cigarettes/day) and in 11 nonsmoking controls matched for age, sex, and body weight. There was no significant difference between controls and cigarette smokers in desmethyldiazepam volume of distribution (1.29 vs 1.34 liters/kg), elimination half-life (55 vs 59 h), clearance (0.284 vs 0.276 ml/min/kg), or free fraction in serum (1.96 vs 1.92% unbound). Thus, propranolol slightly although significantly impairs the clearance of desmethyldiazepam and prolongs its half-life. Cigarette smoking has no apparent influence on desmethyldiazepam kinetics.