Over expression of ubiquitin-conjugating enzyme E2O in bone marrow mesenchymal stromal cells partially attenuates acute myeloid leukaemia progression.
Bone marrow mesenchymal stromal cells (BM-MSCs) are implicated in the pathogenesis of acute myeloid leukaemia (AML). However, due to the high heterogeneity of AML the mechanism underlying the cross-talk between MSCs and leukaemia cells is not well understood. We found that mixed-lineage leukaemia-AF9 (MLL-AF9)-induced AML mice-derived MSCs had higher proliferative viability compared to wild-type mice-derived MSCs with ubiquitin-conjugating enzyme E2O (Ube2o) down-regulation. After overexpression of UBE2O in AML-derived MSCs, the growth capacity of MSCs was reduced with nuclear factor kappa B subunit 1 (NF-κB) pathway deactivation. In vitro co-culture assay revealed that UBE2O-overexpression MSCs suppressed the proliferation and promoted apoptosis of AML cells by direct contact. In vivo results revealed that the leukaemia burden was reduced and the overall survival of AML mice was prolonged, with decreased dissemination of leukaemia cells in BM, spleen, liver and peripheral blood. Additionally, subcutaneous tumorigenesis revealed that tumour growth was also suppressed in the UBE2O-overexpression MSCs group. In conclusion, UBE2O was expressed at a low level in MLL-AF9-induced AML mice-derived MSCs. Overexpression of UBE2O in MSCs suppressed their proliferation through NF-κB pathway deactivation, which resulted in AML suppression. Our study provides a theoretical basis for a BM microenvironment-based therapeutic strategy to control disease progression.


Inhibition of the CCR6-CCL20 axis prevents regulatory T cell recruitment and sensitizes head and neck squamous cell carcinoma to radiation therapy.
Radioresistance of HNSCCs remains a major challenge for effective tumor control. Combined radiotherapy (RT) and immunotherapy (IT) treatment improved survival for a subset of patients with inflamed tumors or tumors susceptible to RT-induced inflammation. To overcome radioresistance and improve treatment outcomes, an understanding of factors that suppress anti-tumor immunity is necessary. In this regard, regulatory T cells (Tregs) are critical mediators of immune suppression in HNSCCs. In this study, we investigated how radiation modulates Treg infiltration in tumors through the chemokine CCL20. We hypothesized that radiation induces CCL20 secretion resulting in Treg infiltration and suppression of anti-tumor immunity. Human and mouse HNSCC cell lines with different immune phenotypes were irradiated at doses of 2 or 10 Gy. Conditioned media, RNA and protein were collected for assessment of CCL20. qPCR was used to determine CCL20 gene expression. In vivo, MOC2 cells were implanted into the buccal cavity of mice and the effect of neutralizing CCL20 antibody was determined alone and in combination with RT. Blood samples were collected before and after RT for analysis of CCL20. Tumor samples were analyzed by flow cytometry to determine immune infiltrates, including CD8 T cells and Tregs. Mass-spectrometry was performed to analyze proteomic changes in the tumor microenvironment after anti-CCL20 treatment. Cal27 and MOC2 HNSCCs had a gene signature associated with Treg infiltration, whereas SCC9 and MOC1 tumors displayed a gene signature associated with an inflamed TME. In vitro, tumor irradiation at 10 Gy significantly induced CCL20 in Cal27 and MOC2 cells relative to control. The increase in CCL20 was associated with increased Treg migration. Neutralization of CCL20 reversed radiation-induced migration of Treg cells in vitro and decreased intratumoral Tregs in vivo. Furthermore, inhibition of CCL20 resulted in a significant decrease in tumor growth compared to control in MOC2 tumors. This effect was further enhanced after combination with RT compared to either treatment alone. Our results suggest that radiation promotes CCL20 secretion by tumor cells which is responsible for the attraction of Tregs. Inhibition of the CCR6-CCL20 axis prevents infiltration of Tregs in tumors and suppresses tumor growth resulting in improved response to radiation.


Drinking in adulthood: similarities and differences in effects of adult roles for Hispanic ethnic groups and Anglos.
This study examines whether the adult social roles perspective. an approach that explains drinking behaviors for Anglos, similarly affects alcohol use by Cubans, Mexican Americans, Puerto Ricans and Other Hispanics (Central and South Americans). The 1993 National Household Survey on Drug Abuse, a national probability sample of the household population in the United States, is used. The sample utilized here (N= 13,822; 56.2% female) consisted of 9,388 Anglos, 611 Cubans, 2,459 Mexican Americans, 611 Puerto Ricans and 753 Central/South Americans age 18 and older. The outcome measures include past-year drinking, and for drinkers, heavy drinking and alcohol-related problems. Logistic regression analyses are conducted using Stata. The results show that there are some ethnic differences in the effects of the adult social roles. Of particular importance is the finding that being married has anomalous effects for Cubans (heavy drinking), Mexican Americans (problems) and Other Hispanics (problems) compared with Anglos and the other Hispanic ethnic groups. The adult social roles perspective has some utility for explaining Hispanic drinking patterns. Future research should consider not only traditional predictors of drinking but also such socio-cultural factors as acculturation and familism, to better understand adult alcohol use by members of Hispanic ethnic groups.


Cuproptosis related gene PDHB is identified as a biomarker inversely associated with the progression of clear cell renal cell carcinoma.
Cuproptosis is a newly discovered programmed cell death dependent on mitochondrial respiratory disorder induced by copper overload. Pyruvate dehydrogenase E1 subunit beta (PDHB) is one of the cuproptosis genesand is a nuclear-encoded pyruvate dehydrogenase, which catalyzes the conversion of pyruvate to acetyl coenzyme A. However, the mechanism of PDHB in clear cell renal cell carcinoma (ccRCC) remains unclear. We used data from TCGA and GEO to assess the expression of PDHB in normal and tumor tissues. We further analyzed the relationship between PDHB and somatic mutations and immune infiltration. Finally, we preliminarily explored the impact of PDHB on ccRCC. The expression level of PDHB was lower in tumor tissue compared with normal tissue. Meanwhile, the expression level of PDHB was also lower in high-grade tumors than low-grade tumors. PDHB is positively correlated with prognosis in ccRCC. Furthermore, PDHB may be associated with decreased risk of VHL, PBRM1 and KDM5C mutations. In 786-O cells, copper chloride could promote the expression of cuproptosis genes (DLAT, PDHB and FDX1) and inhibit cell growth. Last but not least, we found that PDHB could inhibit the proliferation and migration of ccRCC cells. Our results demonstrated that PDHB could inhibit the proliferation, migration and invasion in ccRCC cells, which might be a prognostic predictor of ccRCC. Targeting this molecular might provide a new therapeutic strategy for patients with advanced ccRCC.


Theoretical Thermodynamics Study of Polyamidoamine Deposited Around a Nanotube as Motor Controlled by Light and Under Temperature Effect.
We simulated a system like a Polyamidoamine (PAMAM) deposited on open carbon nanotube. We used five first generation PAMAM. The initial position of PAMAM is out of CN symmetry position. It permits the PAMAM to relax around the nanotube due to van der Waals force. After that, we have analyzed the thermal effects on behavior of 4G PAMAM. We performed computational simulation using classical molecular dynamics with standard parameterization. The thermodynamics properties of this device as molar specific heat and molar entropy variation were calculated. The CN has 690 carbon atoms with up to almost 0.1 ns of simulation.


The development and evolution of lamellar macular hole in highly myopic eyes.
To study the development, evolution, outcomes, and prognostic factors of lamellar macular hole (LMH) in highly myopic (HM) patients. Fifty eyes from 47 HM patients with LMHs were retrospectively enrolled. Relevant pre- and post-LMH optical coherence tomography findings and visual acuity were collected. Structural progression was defined as an increase in the height of retinoschisis, and the development of foveal detachment, full-thickness macular hole, or retinal detachment. Four traction-related developmental processes were identified. Type 1 LMHs (8, 16%) developed from foveal avulsion caused by vitreomacular traction. Type 2 (32, 64%) and type 3 LMHs (5, 10%) formed from ruptured parafoveal and central foveal cysts, respectively. Progressive foveal thinning caused by epiretinal membranes (ERMs) without cystic changes led to type 4 LMHs (5, 10%). Retinoschisis developed before (9 eyes), after (10 eyes), or simultaneously with (6 eyes) the LMH formation. Structural progression was noted in 50%, 53%, 0%, 100% of patients with type 1-4 LMHs, respectively. Multivariable Cox proportional hazard model showed that greater residual foveal thickness (P = 0.001, adjusted odds ratio = 0.22, 95% confidence interval [CI], 0.08 ~ 0.56), and the absence of retinoschisis were protective against structural progression. Multivariable linear regression showed that poor baseline visual acuity (P < 0.001, β = 0.74, 95% CI 0.41 ~ 1.07) and type 4 LMH predicted worse visual outcomes. Four traction-related LMH developmental processes were observed in HM eyes and exhibited different evolution and outcomes. LMHs with foveal thinning induced by ERMs had the worst outcomes.


DNA with Damage in Both Strands as Affinity Probes and Nucleotide Excision Repair Substrates.
Nucleotide excision repair (NER) is a multistep process of recognition and elimination of a wide spectrum of damages that cause significant distortions in DNA structure, such as UV-induced damage and bulky chemical adducts. A series of model DNAs containing new bulky fluoro-azidobenzoyl photoactive lesion dC(FAB) and well-recognized nonnucleoside lesions nFlu and nAnt have been designed and their interaction with repair proteins investigated. We demonstrate that modified DNA duplexes dC(FAB)/dG (probe I), dC(FAB)/nFlu+4 (probe II), and dC(FAB)/nFlu-3 (probe III) have increased (as compared to unmodified DNA, umDNA) structure-dependent affinity for XPC-HR23B (Kdum > KdI > KdII ≈ KdIII) and differentially crosslink to XPC and proteins of NER-competent extracts. The presence of dC(FAB) results in (i) decreased melting temperature (ΔTm = -3°C) and (ii) 12° DNA bending. The extended dC(FAB)/dG-DNA (137 bp) was demonstrated to be an effective NER substrate. Lack of correlation between the affinity to XPC-HR23B and substrate properties of the model DNA suggests a high impact of the verification stage on the overall NER process. In addition, DNAs containing closely positioned, well-recognized lesions in the complementary strands represent hardly repairable (dC(FAB)/nFlu+4, dC(FAB)/nFlu-3) or irreparable (nFlu/nFlu+4, nFlu/nFlu-3, nAnt/nFlu+4, nAnt/nFlu-3) structures. Our data provide evidence that the NER system of higher eukaryotes recognizes and eliminates damaged DNA fragments on a multi-criterion basis.


Facilitation of stimulus detection performance of rats with d-amphetamine: a function of dose and level of training.
Conditions under which amphetamine may facilitate stimulus detection task choice performance in rats were investigated. Rats (n=15) were trained in a two-choice, light-detection task to three successively more stringent criterion levels of task training (minimal, intermediate, and extended) and then tested after administration of saline, 0.25, 0.50, and 0.75 mg/kg d-amphetamine (AMP). For each training level, baseline levels of choice accuracy were maintained at approximately 82% by manipulating the animals' cue duration. No aspect of performance was enhanced by any dose of AMP after minimal criteria training, and there was a dose-dependent decrease in the number of trials completed. After the intermediate level of training, the 0.25 mg/kg dose of AMP reliably increased choice accuracy, there was no reliable change in choice reaction time, and there was a dose-dependent decrease in the number of trials completed. After the extended training, the 0.25 mg/kg dose of AMP reliably increased choice response accuracy, the 0.25 and 0.50 mg/kg doses of AMP reliably decreased choice reaction time, and there was no reliable change in the number of trials completed at any dose of AMP. These results support the contention that psychostimulants can facilitate the choice performance of rats in stimulus detection tasks if an appropriately low dose is used and the animal's behavior is strongly controlled by the stimulus-reinforcement contingencies of the task.


The impact of the SARS-CoV-2 pandemic-related lockdowns on orthopedic trauma emergencies at a level-one trauma center.
The SARS-CoV-2 pandemic and its associated lockdowns had a profound effect on orthopedic trauma emergencies. This study aimed to investigate the patient volume and injury patterns at a level-one trauma center during the SARS-CoV-2 pandemic and compare them to the pre-pandemic conditions. A retrospective chart review of all patients who presented to the orthopedic trauma emergency department of a level-one trauma center in Cologne, Germany within a 2 year period from March 16th, 2019 to March 15th, 2020 (pre-pandemic control) and from March 16th, 2020 and March 15th, 2021 (pandemic) was performed. The pandemic year was separated into three periods: (1) first lockdown, (2) between lockdowns and (3) second lockdown. The absolute numbers of patient presentations, the Manchester triage score (MTS) and the relative proportion of patients with structural organ injuries, fractures and dislocations, of polytraumatized patients, of hospital admissions, of subsequent emergency or semi-elective surgeries and of work-related accidents were evaluated in comparison to the pre-pandemic control. A total of 21,642 patient presentations were included in this study. Significantly less weekly orthopedic trauma emergency patient presentations were recorded during the pandemic (p < 0.01). The MTS was significantly lower during the first lockdown and between lockdowns (p < 0.01). The proportional incidence of overall structural organ injuries, fractures and dislocations, of upper limb fractures/dislocations, of hospital admissions and of patients requiring surgery was significantly increased during the pandemic (p ≤ 0.03). The proportional incidence of work-related injuries was significantly decreased during the pandemic (p < 0.01). Orthopedic trauma emergency presentations were reduced during the SARS-CoV-2 pandemic. Due to the reluctancy of patients to visit the emergency department during the pandemic, the proportions of relevant injuries in general and of upper limb injuries in particular as well as of patients requiring hospital admission and trauma-related surgery were significantly increased.


Monoclonal Antibodies Hypersensitivity: Prevalence and Management.
The use of monoclonal antibodies (mAbs) has become broader because of their recognized effectiveness in the treatment of autoimmune, neoplastic, and inflammatory diseases. Consequently, hypersensitivity reactions (HSR) secondary to mAbs are being reported more often, and each mAb-related HSR presents specific features. This article discusses the main biological agents and associated HSR, the clinical presentation of such reactions, and the role of tryptase and skin testing in the diagnosis. Rapid drug desensitization procedures to mAbs enable selected allergic patients to receive full therapeutic doses in a safe manner and are also discussed.


A cross-sectional study examining MRCS performance by core surgical training location.
In the UK, core surgical training (CST) is the first specialty experience that early-career surgeons receive but training differs significantly across CST deaneries. To identify the impact these differences have on trainee performance, we assessed whether success at the Membership of the Royal College of Surgeons (MRCS) examinations is associated with CST deanery. A retrospective cohort study of UK trainees in CST who attempted MRCS between 2014 and 2020 (n = 1104). Chi-squared tests examined associations between locality and first-attempt MRCS performance. Multivariate logistic regression models identified the likelihood of MRCS success depending on CST deanery. MRCS Part A and Part B pass rates were associated with CST deanery (p < 0.001 and p = 0.013, respectively). Candidates that trained in Thames Valley (Odds Ratio [OR] 2.52 (95% Confidence Interval [CI] 1.00-6.42), North Central and East London (OR 2.37 [95% CI 1.04-5.40]) or South London (OR 2.36 [95% CI 1.09-5.10]) were each more than twice as likely to pass MRCS Part A at first attempt. Trainees from North Central and East London were more than ten times more likely to pass MRCS Part B at first attempt (OR 10.59 [95% CI 1.23-51.00]). However, 68% of candidates attempted Part A prior to CST and 48% attempted Part B before or during the first year of CST. MRCS performance is associated with CST deanery; however, many candidates passed the exam with little or any CST experience suggesting that some deaneries attract high academic performers. MRCS performance is therefore not a suitable marker of CST training quality.


The Appropriateness of Preoperative Patient Reported Outcome Measures as an Indication for Total Hip Arthroplasty.
While Medicare requires patient-reported outcome measures (PROMs) for many quality programs, some commercial insurers have begun requiring preoperative PROMs when determining patient eligibility for total hip arthroplasty (THA). Concerns exist these data may be used to deny THA to patients above a specific PROM score, but the optimal threshold is unknown. We aimed to evaluate outcomes following THA based on theoretical PROM thresholds. We retrospectively analyzed 18,006 consecutive primary THA patients from 2016-2019. Hypothesized preoperative Hip Disability and Osteoarthritis Outcome Score, Joint Replacement (HOOS-JR) cutoffs of 40, 50, 60, and 70 points were used. Preoperative scores below each threshold were considered "approved" surgery. Preoperative scores above each threshold were considered "denied" surgery. In-hospital complications, 90-day readmissions, and discharge disposition were evaluated. HOOS-JR scores were collected preoperatively and 1-year postoperatively. Minimum clinically important difference (MCID) achievement was calculated using previously validated anchor-based methods. Using preoperative HOOS-JR thresholds of 40, 50, 60, and 70 points, the percentage of patients who would have been denied surgery was 70.4%, 43.2%, 20.3%, and 8.3%, respectively. For these denied patients, 1-year MCID achievement was 75.9%, 69.0%, 59.1%, and 42.1%, respectively. In-hospital complication rates for approved patients were 3.3%, 3.0%, 2.8%, and 2.7%, while 90-day readmission rates were 5.1%, 4.4%, 4.2%, and 4.1%, respectively. Approved patients had higher MCID achievement (P < .001) but higher nonhome discharge (P = .01) and 90-day readmissions rates (P = .036) than denied patients. Most patients achieved MCID at all theoretical PROM thresholds with low complication and readmission rates. Setting preoperative PROM thresholds for THA eligibility did not guarantee clinically successful outcomes.


Recovery of Angiostrongylus cantonensis from cerebrospinal fluid of a child with eosinophilic meningitis.
Viable Angiostrongylus cantonensis was recovered from the cerebrospinal fluid of a 17-month-old boy with eosinophilic meningitis. Neurological findings were minimal, and the child had an uneventful recovery.


Can early improvement be an indicator of treatment response in obsessive-compulsive disorder? Implications for early-treatment decision-making.
In major depression, early response to treatment has been strongly associated with final outcome. We aimed to investigate the ability of early improvement (4 weeks) to predict treatment response at 12 weeks in DSM-IV-defined obsessive-compulsive disorder (OCD) patients treated with serotonin reuptake inhibitors (SRI). We conducted an SRI practical trial with 128 subjects. age range 18-65 years-old, baseline Yale-Brown Obsessive-Compulsive Scale (Y-BOCS) score ≥ 16, and absence of previous adequate pharmacological treatment. Systematic assessments were performed at baseline, 4 and 12 weeks of treatment. Treatment response at 12 weeks was defined as a 35% or greater reduction in baseline Y-BOCS score. Stepwise logistic regression was used to test the relationship between early improvement and treatment response at 12 weeks, taking into account additional potential predictive factors. Different thresholds of early improvement were tested and their predictive power was calculated. Early improvement, defined as a 20% or greater reduction from baseline Y-BOCS score at 4 weeks, predicted response at 12 weeks with 75.6% sensitivity and 61.9% specificity. According to a logistic regression including demographic and clinical features as explaining variables, early improvement was the best predictor of treatment response (OR = 1.05, p < 0.0001). Only 19.8% of patients who did not improve at 4 weeks were responders after 12 weeks. In contrast, 55.3% of the individuals who showed early improvement were responders at 12 weeks (Pearson Chi-Square = 17.06, p < 0.001). Early improvement predicted OCD treatment response with relatively good sensitivity and specificity, such that its role in early decision-making warrants further investigation in wider samples. clinicaltrials.gov Identifier NCT00680602.


Enzyme immunoassay and enzyme-linked fluoroimmunoassay for detection of Chlamydia trachomatis antigen.
An enzyme immunoassay (EIA) and an enzyme-linked fluoroimmunoassay (ELFIA) utilizing monoclonal antibody to major outer membrane protein of Chlamydia trachomatis L2 were developed for rapid detection of chlamydial antigen in clinical samples. The EIA and ELFIA could detect levels of purified chlamydial outer membrane protein as low as 1.0 and 0.2 ng/ml respectively. However, when EIA and ELFIA were compared to chlamydial isolation using 160 patient samples, the sensitivity rate was 68% and 85% respectively. The sensitivity of the antigen detection method might be increased by simply using less diluted samples than in the present study. Chlamydial antigen was also demonstrated by EIA and ELFIA in 15% and 23% of culture-negative samples. The reason for these false-positive findings remains undetermined.


Event-specific interventions to minimize COVID-19 transmission.
COVID-19 is a global pandemic with over 25 million cases worldwide. Currently, treatments are limited, and there is no approved vaccine. Interventions such as handwashing, masks, social distancing, and "social bubbles" are used to limit community transmission, but it is challenging to choose the best interventions for a given activity. Here, we provide a quantitative framework to determine which interventions are likely to have the most impact in which settings. We introduce the concept of "event R," the expected number of new infections due to the presence of a single infectious individual at an event. We obtain a fundamental relationship between event R and four parameters: transmission intensity, duration of exposure, the proximity of individuals, and the degree of mixing. We use reports of small outbreaks to establish event R and transmission intensity in a range of settings. We identify principles that guide whether physical distancing, masks and other barriers to transmission, or social bubbles will be most effective. We outline how this information can be obtained and used to reopen economies with principled measures to reduce COVID-19 transmission.


Interference with anoikis-induced cell death of dopamine neurons: implications for augmenting embryonic graft survival in a rat model of Parkinson's disease.
One promising therapy for the treatment of Parkinson's disease is transplantation of embryonic ventral mesencephalic tissue. Unfortunately, up to 95% of grafted cells die, many via apoptosis. In this study we attempted to prevent anoikis-induced cell death, which is triggered during the preparation of cells for grafting, and examine the impact on graft viability and function. We utilized the extracellular matrix molecule tenascin-C (tenascin) and an antibody (Ab) to the cell adhesion molecule L1 to specifically mimic survival signals induced by cell-matrix and cell-cell interactions. In vitro, both tenascin- and L1 Ab-treated cultures doubled the number of tyrosine hydroxylase immunoreactive (THir) neurons compared to control. Additionally, cell survival assays determined that tenascin and L1 Ab-treated cell suspensions yielded more metabolically active and fewer dead cells than control suspensions. In contrast to the culture results, tenascin- and L1 Ab-treated mesencephalic grafts did not yield an increase in the number of THir neurons using our standard grafting paradigm (3 microl of 100,000 cells/microl). However, under low-density conditions (3 microl of 3,000 cells/microl), tenascin augmented grafted THir neuron survival. These findings are consistent with the view that cell density can dramatically influence the degree of stress placed on THir neurons and consequently affect the success of survival strategies in vivo. In conclusion, pretreatment with tenascin may prove beneficial to prevent anoikis in dilute cell suspension grafts, while long-term in vivo delivery methods need to be explored to determine if L1 can prevent anoikis in grafts of mesencephalic dopamine neurons after transplantation.


Persistent exercise fatigue and associative learning deficits in combination with transient glucose dyshomeostasis in a mouse model of Gulf War Illness.
To characterize exercise fatigue, metabolic phenotype and cognitive and mood deficits correlated with brain neuroinflammatory and gut microbiome changes in a chronic Gulf War Illness (GWI) mouse model. The latter have been described in an accompanying paper [1]. Adult male C57Bl/6N mice were exposed for 28 days (5 days/week) to pyridostigmine bromide: 6.5 mg/kg, b.i.d., P.O. (GW1) or 8.7 mg/kg, q.d., P.O. (GW2); topical permethrin (1.3 mg/kg in 100% DMSO) and N,N-diethyl-meta-toluamide (DEET 33% in 70% EtOH) and restraint stress (5 min). Exercise, metabolic and behavioral endpoints were compared to sham stress control (CON/S). Relative to CON/S, GW2 presented persistent exercise intolerance (through post-treatment (PT) day 161), deficient associative learning/memory, and transient insulin insensitivity. In contrast to GW2, GW1 showed deficient long-term object recognition memory, milder associative learning/memory deficit, and behavioral despair. Our findings demonstrate that GW chemicals dose-dependently determine the presentation of exercise fatigue and severity/type of cognitive/mood-deficient phenotypes that show persistence. Our comprehensive mouse model of GWI recapitulates the major multiple symptom domains characterizing GWI, including fatigue and cognitive impairment that can be used to more efficiently develop diagnostic tests and curative treatments for ill Gulf War veterans.


An experimental demonstration of density-dependent reproduction in a natural population of Diamorpha smallii, a rare annual.
In a North Carolina population of Diamorpha smallii, densities were artificially manipulated by thinning stands to predetermined densities. Individuals in areas of high density produce fewer seeds than individuals at low density in the same habitat. Three components of fecundity, the number of flowers per plant, the number of fruits per plant, and the number of seeds per fruit, were negatively correlated with density. The relationship of components of fecundity to density were used to develop a model of population regulation for this species. These results and predictions from the model are discussed in relation to other density studies.


Biochemical methods for studying kinetic regulation of Arf1 activation by Sec7.
The ADP ribosylation factor (Arf) family of small guanosine triphosphatases (GTPases) regulates vesicular transport at several locations within the cell, and is in turn regulated by guanine nucleotide exchange factors (GEFs) via a conserved catalytic domain, termed the Sec7 domain. The catalytic activity of the Sec7 domain is well characterized in the context of a few GEFs acting at the periphery of the cell. This chapter describes the techniques used to extend the biochemical analysis of activity to the much larger GEFs acting on the Arf family in the core secretory pathway, using the activity of Saccharomyces cerevisiae Sec7 on Arf1, regulating export from the trans-Golgi network, as a model. The complete methods for purification to near homogeneity of all proteins required, including several Sec7 constructs and multiple relevant small GTPases, are detailed. These are followed by methods for the quantification of the nucleotide exchange activity of Sec7 in a physiologically relevant context, including modifications required to dissect the signal integration functions of Sec7 as an effector of several other small GTPases, and methods for identifying stable Sec7-small GTPase interactions in the presence of membranes. These techniques may be extended to the analysis of similar members of the Sec7 GEF subfamily in other species and acting elsewhere in the secretory pathway.


The role of natural killer T cells in B cell malignancies.
Little is known regarding the precise role of different subsets of natural killer T (NKT) cells in the immunopathogenesis of cancer diseases, particularly hematopoietic malignancies. Although it is well known that NKT cells counteract tumor immunity, conflicting reports on the role of NKT cells in hematopoietic malignancies support more investigations to clarify the interactions between NKT cells and the tumor. Among the hematopoietic malignancies, B cell malignancies derive from different stages of B cell maturation in which T cells play a pivotal role. There is evidence which implies the protective role of some subsets of NKT cells in solid cancers as well as B cell malignancies. In this review, we will discuss recent advances about the immunobiology of NKT cells and their precise role in the immunopathogenesis and treatment of different B cell malignancies.


Mesenchymal stromal cells protect mantle cell lymphoma cells from spontaneous and drug-induced apoptosis through secretion of B-cell activating factor and activation of the canonical and non-canonical nuclear factor κB pathways.
There is increasing evidence that stromal cell interactions are required for the survival and drug resistance of several types of B-cell malignancies. There is relatively little information regarding the role of the bone marrow/lymphoid microenvironment in the pathogenesis of mantle cell lymphoma. In this study we investigated the interaction of primary mantle cell lymphoma cells with stromal cells in an ex vivo co-culture system. The murine stromal cell line MS-5 and human bone marrow mesenchymal stromal cells were each co-cultured with primary mantle cell lymphoma cells for up to 7 months. Mantle cell lymphoma cultures alone or combined with human stromal cells were analyzed for cell number, cell migration, nuclear factor-κB activation and drug resistance. Co-culture of mantle cell lymphoma cells and human stromal cells results in the survival and proliferation of primary mantle cell lymphoma cells for at least 7 months compared to mantle cell lymphoma cells cultured alone. Mantle cell lymphoma-human stromal cell interactions resulted in activation of the B-cell activating factor/nuclear factor-κB signaling axis resulting in reduced apoptosis, increased mantle cell lymphoma migration and increased drug resistance. Direct mantle cell lymphoma-human stromal cell interactions support long-term expansion and increase the drug-resistance of primary mantle cell lymphoma cells. This is due in part to activation of the canonical and non-canonical nuclear factor κB pathways. We also demonstrated the ability of B-cell activating factor to augment CXCL12- and CXCL13-induced cell migration. Collectively, these findings demonstrate that human stromal cell-mantle cell lymphoma interactions play a pivotal role in the pathogenesis of mantle cell lymphoma and that analysis of mantle cell lymphoma-human stromal cell interactions may help in the identification of novel targets for therapeutic use.


Phenotypic features of a boy with trisomy of 16q22-->qter due to paternal Y; 16 translocation.
We report on the phenotypic features of a patient with partial trisomy of the long arm of chromosome 16 due to an unbalanced Y;16 translocation (46,X,der[Y]t[Y;16] [q12;q22]pat). The patient was noted to have craniofacial anomalies and developmental delay, but no other major malformations. The father, a balanced Y;16 translocation carrier, has apparently normal fertility.


Rest and exercise performance of the Medtronic Advantage bileaflet valve in the aortic position.
The aim of the study was to evaluate rest and exercise performance and left ventricular mass regression of the Medtronic Advantage (Medtronic, Inc, Minneapolis, MN) prosthesis in the aortic position at 1 year at a single center as part of a multicenter, prospective clinical trial. Between May 2002 and June 2003, 63 consecutive patients underwent aortic valve replacement with a Medtronic Advantage prosthesis (84.1% male; mean age, 56.0 +/- 9.7 years; ejection fraction, 56.5 +/- 15.8%). Valve lesions were stenosis (n = 20), mixed (n = 34), and insufficiency (n = 9). Concomitant procedures were performed in 34.9%. Follow-up was 100% complete. Echocardiographic data were obtained early postoperatively and at 1 year, combined with stress echocardiography by treadmill. Mean pressure gradients, stroke volume, and left ventricular mass were determined by echocardiography. Data are presented as mean +/- standard deviation. Operative mortality was 0%. Valve-related complications were observed in 2 patients (endocarditis, n = 1; thromboembolic event, n = 1). There was no case of antithromboembolic hemorrhage, prosthesis-related explant, or reoperation. One patient showed moderate paravalvular regurgitation. Mean pressure gradients 1 year postoperatively ranged from 6.3 to 11.0 mm Hg across all valve sizes. Left ventricular mass regression at 1 year was 18.4% across all valve sizes (p < 0.001). No severe patient-prosthesis mismatch (effective orifice area index < or = 0.65 cm2/m2) could be observed. After 1 year, the Medtronic Advantage valve shows comparable transvalvular mean pressure gradients across the valve sizes used during rest and exercise. This is accompanied by a significant left ventricular mass regression, an important indicator for long-term survival.


Response of irrigated tomato (Solanum lycopersicum Mill) to mulch application rates.
Rapid decline in available water for crop production has led to the adoption of irrigation schedules for meeting water supply throughout cropping seasons. Nonetheless, the loss of water from soil often results in spells of water stress between schedules, which adversely affect crop yield. Hence, the use of mulch in conserving soil moisture in irrigated farming is becoming popular among farmers. In this study, a two-year screenhouse pot experiment was conducted to evaluate the effects of Pennisetum purpureum (Pp) mulch on tomato (Roma variety) grown in daily irrigation (IFdaily), irrigation at 3-days interval (IF3), and irrigation at 5-days interval (IF5) conditions. The Pp mulch was chopped to 5 cm and applied on the soil surface of each experimental pot at 1 t ha-1 (Pp1), 2 t ha-1 (Pp2), 3 t ha-1 (Pp3), and 4 t ha-1 (Pp4). These rates were compared against a bare soil as control (Pp0). The treatments were laid in a completely randomised design with four replicates. Tomato yield decreased by 53.6% and 26.6% in IF3, and 86.2% and 65.0% in IF5 compared with IFdaily in years 1 and 2, respectively. Among mulch rates, Pp4 and Pp3 increased tomato yield respectively by 107.5% and 99.9% compared with Pp0, while Pp2 and Pp1 were similar in year 1. In year 2, mulch increased tomato yield by 84.1% (Pp1) - 215.3% (Pp4) and contributed substantially to tomato yield in IFdaily (R2 = 0.99; p < 0.01); IF3 (R2 = 0.93; p < 0.01); and IF5 (R2 = 0.25; p < 0.05). However, withdrawing irrigation at 5 days interval was detrimental to tomato yield production.


Two unrelated cases of single maxillary central incisor with 7q terminal deletion.
Two unrelated cases of single maxillary central incisor (SM-CI) with 7q terminal deletion of the same breakpoint at 7q36.1 were described. They had mental retardation, microcephaly, hypotelorism, short stature, and normal levels of plasma growth hormone. One case had bilateral caudal ectopic kidneys, double renal pelves, and dilated ureters. The other had bilateral hydroureteronephrosis. The present cases suggest that 7q terminal deletion is one of the causes of SMCI.


Conformational transitions in the synthetic polynucleotide poly[d(G-C)] . poly[d(G-C)] double-helix.
Conditions are described for observing by X-ray fibre diffraction the A, B and S conformations of the poly[d(G-C)] . poly[d(G-C)] double-helix and also a new form designated as B". For fibres with an appropriate ionic content, transitions between these conformations can be induced by varying the relative humidity of the fibre environment. With increasing relative humidity the transitions B" leads to A leads to S leads to B occur. However, reducing the relative humidity does not result in a simple reversal of these transitions. If the relative humidity is reduced rapidly, a B leads to A transition is observed followed by an A leads to B" transition, but if it is reduced slowly, the transition is from B to S. Once the S form has been assumed, further reduction in the relative humidity does not result in a transition to the A form. The S form emerges as a particularly stable form of the poly[d(G-C)] . poly[d(G-C)] double-helix. From the point of view of its relationship to the classical A and B forms, the S form of poly[d(G-C)] . poly[d(G-C)] is shown to exhibit similarities to the D form of poly[d(A-T)] . poly[d(A-T)].


Correction of a genetic defect by nuclear transplantation and combined cell and gene therapy.
Immune-deficient Rag2(-/-) mice were used as nuclear donors for transfer into enucleated oocytes, and the resulting blastocysts were cultured to isolate an isogenic embryonic stem cell line. One of the mutated alleles in the Rag2(-/-) ES cells was repaired by homologous recombination, thereby restoring normal Rag2 gene structure. Mutant mice were treated with the repaired ES cells in two ways. (1) Immune-competent mice were generated from the repaired ES cells by tetraploid embryo complementation and were used as bone marrow donors for transplantation. (2) Hematopoietic precursors were derived by in vitro differentiation from the repaired ES cells and engrafted into mutant mice. Mature myeloid and lymphoid cells as well as immunoglobulins became detectable 3-4 weeks after transplantation. Our results establish a paradigm for the treatment of a genetic disorder by combining therapeutic cloning with gene therapy.


The effects of fluoride concentration and the level of cariogenic challenge on caries development in desalivated rats.
Dental caries is an infectious and transmissible disease that continues to affect the majority of people. The presence of carbohydrate, mainly sucrose in the diet, is an important factor in its occurrence. The amount of fluoride required for optimal protective effect where there is a high caries challenge is unclear. Differences in the intensity of cariogenic challenge, for whatever reason, may play a part in determining fluctuations in the effectiveness of fluoride. The purpose of this study was to evaluate the effect of different concentrations of fluoride on the development of caries and explore the cariostatic effect of fluoride under various levels of cariogenic challenge. The study comprises two experiments. In experiment I, 60 desalivated Sprague Dawley rats infected with Streptococcus sobrinus were offered the following to drink for 21 days: group (1), sterile distilled water (SDW); (2) 10 parts/10(6) F SDW; (3) 20 parts/10(6) F SDW; (4) 30 parts/10(6) F SDW; (5) 40 parts/10(6) F SDW. In experiment II, eight groups of 9 rats were placed in a König Höfer programmed feeder and were exposed to different levels of cariogenic challenge through varying frequency of eating and offered water containing 10 parts/10(6) F. In experiment I, exposure to 20, 30 and 40 parts/10(6) F reduced caries development significantly: fluoride, at 10 parts/10(6), reduced the severity of the carious lesions. In this model of severe cariogenic challenge, the results suggest that elevated concentrations of fluoride might be effective in patients at high caries risk. In experiment II, fluoride reduced the incidence and severity of smooth-surface caries in all groups. The protective effect of fluoride decreased as the number of exposures to sugar increased. It is concluded that the effectiveness of fluoride is influenced by the level of cariogenic challenge and that consideration should be given to adjusting the level of fluoride exposure based on perceived caries risk, and that there is a maximum therapeutic effect of fluoride beyond which no additional protection can be expected.


Cardiovascular Risk Factors in Obese Children and Adolescents.
The aim of the study was to analyze cardiometabolic risk factors andcarotid intima-media thickness (IMT) in obese children. We studied 122 obese children fulfilling the criteria of the International Obesity Task Force and 58 non-obese children. Anthropometric parameters, blood pressure, lipid profile, C-reactive protein, and adiponectin were assessed in all children. Glucose and insulin during the oral glucose tolerance test were assessed in obese children. The IMT was determined using ultrasound B-mode imaging in 81 obese and 32 non-obese children. We found that obese children had significantly higher levels of lipid andother non-lipid atherogenic indicators, but lower levels of adiponectin compared with non-obese children. The difference in the mean carotid IMT was insignificant in the two groups. Taking the combined groups, the level of adiponectin correlated negatively with body mass index and lipid atherogenic indicators. The IMT strongly correlated with systolic blood pressure in obese children. In the children fulfilling the criteria of metabolic syndrome, 17 out of the 84 obese children older than 10 years of age, IMT was greater than in those who did not fulfil these criteria. We conclude that the coexistence of abdominal obesity with abnormal lipid profile and hypertension leads to the early development of atherosclerosis accompanied by increased carotid intima-media thickness. Obesity initiates the atherosclerotic processes in early childhood.


Nonvolatile Electrical Bistability Behaviors Observed in Au/Ag Nanoparticle-Embedded MOFs and Switching Mechanisms.
Electrically bistable devices play an important role in the next generation of information materials. Plasmonic noble metal nanoparticles (Au and Ag NPs) with diameters <6 nm were embedded into 3-D Cd-based metal-organic framework (MOF) matrixes via the photoreduction method to generate Au (Ag) NPs@MOF composites. Electrical bistability measurements on the sandwiched ITO/NPs@MOF/silver devices indicate that two switchable conductivity states with nonvolatile memory behaviors can be observed. The ITO/Au NP@2/Ag device with neutral matrix possesses the highest ON/OFF current ratio of 104, which can be attributed to its higher electron tunneling efficiency because of the better dispersity of Au NPs in the MOF matrix. A mechanism regarding the electric-field-induced charge-transfer process assisted by conformational change in the active layer was proposed.


Inhibition of the thyroid hormonogenic H2O2 production by Duox/DuoxA in zebrafish reveals VAS2870 as a new goitrogenic compound.
Thyroid hormone (TH) synthesis requires extracellular hydrogen peroxide generated by the NADPH oxidases, DUOX1 and DUOX2, with maturation factors, DUOXA1 and DUOXA2. In zebrafish, only one duox and one duoxa gene are present. Using a thyroid-specific reporter line, we investigated the role of Duox and Duoxa for TH biosynthesis in zebrafish larvae. Analysis of several zebrafish duox and duoxa mutant models consistently recovered hypothyroid phenotypes with hyperplastic goiter caused by impaired TH synthesis. Mutant larvae developed enlarged thyroids and showed increased expression of the EGFP reporter and thyroid functional markers including wild-type and mutated duox and duoxa transcripts. Treatment of zebrafish larvae with the NADPH oxidase inhibitor VAS2870 phenocopied the thyroid effects observed in duox or duoxa mutants. Additional functional in vitro assays corroborated the pharmacological inhibition of Duox activity by VAS2870. These data support the utility of this new experimental model to characterize endocrine disruptors of the thyroid function.


Plasminogen activator from cells transformed by an oncogenic virus: inhibitors of the activation reaction.
This paper describes an assay for direct measurement of plasminogen activation and its application for determining the kinetic constants and for screening potential inhibitors of the reaction. The assay is based on the conversion of the single chain of 125I-labelled plasminogen to the two chains of 125I-labelled plasmin (EC 126.96.36.199), the latter then being separated from each other and from the plasminogen substrate by electrophoresis under reducing conditions in SDS-polyacrylamide gels. The Km of activator from transformed murine cells for human plasminogen was 180 nM. A broad range of compounds was tested as potential inhibitors of plasminogen activation and of plasmin-catalyzed fibrinolysis respectively, and the two reactions differed qualitatively and quantitatively in their response to previous agents. The principal qualitative difference was in the susceptibility of the reactions to a spectrum of naturally-occurring macromolecular inhibitors: all of the macromolecular inhibitors that blocked the action of plasmin were without effect on murine activator or human urokinase (EC 184.108.40.206). A variety of small molecules inhibited both of the reactions tested, and showed significant quantitative differences; some of these were active at micron concentrations. The exacting specificity of plasminogen activators for macromolecules, both substrates and inhibitors, encourages the expectation that effective inhibitors of great specificity may be isolated from as yet undiscovered natural sources.


Effects of fear of cancer recurrence on subjective physical and mental health in breast cancer patients: The intermediary role of heart rate variability.
Fear of cancer recurrence (FCR) and psychological distress are common psychological problems in breast cancer (BC) patients and ultimately affecting their health-related quality of life (HRQoL). Heart rate variability (HRV) can reflect the activity of the parasympathetic nervous system. However, the pathways through which HRV influences between FCR and HRQoL are unclear. This study preliminarily explored the intermediary role of HRV in FCR and HRQoL in BC patients. A total of 101 BC patients participated in this study. HRV parameters were measured by a 5-min dynamic electrocardiogram. FCR, psychological distress and HRQoL were evaluated by the Fear of disease progression simplified scale (FOP-Q-SF), Distress thermometer and SF-36 concise health survey. The intermediary effect model was established to test the intermediary effect of high frequency-HRV (HF-HRV) on FCR and HRQoL. FCR and psychological distress were negatively correlated with HRV in the time domain, negatively correlated with HF-HRV in the frequency domain, and positively correlated with low frequency/high frequency (LF/HF). HF-HRV had a partial mediating effect on the FCR and physical health and mental health, with effects of 30.23% and 9.53%, respectively. FCR and psychological distress are correlated with HRV parameters in the time domain and the frequency domain, and we preliminarily believe that parasympathetic nerves play an important intermediary role between FCR and subjective physical and mental health. This may provide intervention information for improving the HRQoL of BC patients.


36-month LipiDiDiet multinutrient clinical trial in prodromal Alzheimer's disease.
The LipiDiDiet trial investigates the effects of the specific multinutrient combination Fortasyn Connect on cognition and related measures in prodromal Alzheimer's disease (AD). Based on previous results we hypothesized that benefits increase with long-term intervention. In this randomized, double-blind, placebo-controlled trial, 311 people with prodromal AD were recruited using the International Working Group-1 criteria and assigned to active product (125 mL once-a-day drink) or an isocaloric, same tasting, placebo control drink. Main outcome was change in cognition (Neuropsychological Test Battery [NTB] 5-item composite). Analyses were by modified intention-to-treat, excluding (ie, censoring) data collected after the start of open-label active product and/or AD medication. Of the 382 assessed for eligibility, 311 were randomized, of those 162 participants completed the 36-month study, including 81 with 36-month data eligible for efficacy analysis. Over 36 months, significant reductions in decline were observed for the NTB 5-item composite (-60%; between-group difference 0.212 [95% confidence interval: 0.044 to 0.380]; P = 0.014), Clinical Dementia Rating-Sum of Boxes (-45%; P = 0.014), memory (-76%; P = 0.008), and brain atrophy measures; small to medium Cohen's d effect size (0.25-0.31) similar to established clinically relevant AD treatment. This multinutrient intervention slowed decline on clinical and other measures related to cognition, function, brain atrophy, and disease progression. These results indicate that intervention benefits increased with long-term use.


Effect of deficit irrigation on physiological, biochemical, and yield characteristics in three baby corn cultivars (Zea mays L.).
The main problem in the production of crops in arid and semi-arid regions of the world is the lack of water and its effect on the plant in the form of drought stress. Cultivation of key crops such as corn, which also requires a lot of water, is not possible in these areas except by applying water consumption management methods. Among the most important of these methods is deficit irrigation. The effect of deficit irrigation on relative water content (RWC), malondialdehyde (MDA), compatible osmolytes (proline and soluble sugars), antioxidant enzymes, and yield was studied in three baby corn cultivars in a field experiment using a randomized complete block design (RCBD) with split-plots and three replications. Three levels of deficit irrigation (0, 20, and 40% deficit) constituted the main plots and three cultivars of baby corn (Challenger, Basin, and Passion) constituted the sub plots. Analysis of variance showed that deficit irrigation had a significant effect on all variables. Cultivar (Challenger, Basin and Passion) had a significant effect on proline (0%, 41.5% and 73.2%), carbohydrates (23.9%, 15.4% and 0%), and MDA content (0%, 26.1% and 41.2%), as well as peroxidase (POD) (0%, 136.1% and 227.9%) levels respectively. The interaction between deficit irrigation and cultivar had a significant effect on proline, carbohydrates, and POD. RWC decreased (26.9, 6.5 and 0%) with increasing irrigation deficit (0, 20 and 40%) respectively while proline (0, 23.7 and 64.8%), carbohydrates (0, 29.7 and 34.09%), catalase (CAT) (0, 20.8 and 70.1%), and POD (0, 55.05 and 113.2%) increased under the same conditions. Carbohydrate content was higher in the Basin and Challenger cultivars (21.71 and 19.07) and proline (145.9), POD (193.9), and MDA content (8.53) were higher in the Passion cultivar. Among the studied cultivars, the highest yield was achieved by the Passion cultivar (37.02 and 62.9% more than Challenger and Basin cultivars respectively). In general, the results showed that drought stress caused an increase in compatible osmolyte content and the activity of antioxidant enzymes. However, this increase could not offset the effects of drought stress on yield in the 40% deficit treatment.


Right ventricular hypertrophy in apical hypertrophic cardiomyopathy.
Right ventricular hypertrophy (RVH+) in hypertrophic cardiomyopathy occurs in one third of patients, however, outcomes in apical hypertrophic cardiomyopathy (ApHCM) have not been described. We hypothesized that RVH+ in ApHCM is associated with more ventricular remodeling and dysfunction, and increased adverse events when compared with those without RVH (RVH-). Ninety-one ApHCM patients were retrospectively analyzed using 2D and speckle-tracking echocardiography (64 ± 16 years old, 43% female). RVH+ was defined as wall thickness >5 mm and was present in 23 (25%). Ventricular mechanics were characterized by global longitudinal strain (GLS), RV free wall strain, and myocardial work. New York Heart Association functional class > II, atrial fibrillation, and prior stroke were more prevalent in RVH+. Left ventricular (LV) size and ejection fraction were similar between groups, with greater septal (17 vs. 14 mm, p = .001) and apical (20 vs. 18 mm, p = .04) wall thickness in RVH+. When compared with RVH- patients, RVH+ had worse LV GLS (-8.6 vs. -12.8%), global work index (820 vs. 1172 mmHg%) (both p < .001), and work efficiency (76 vs. 83%, p = .001), as well as RV GLS (-14 vs. -17.5%) and free wall strain (-17.3 vs. -21.3%) (both p = .02). At 3-year follow-up RVH+ had greater incidence of heart failure hospitalization compared with RVH- (35 vs. 7%, p = .003). RVH+ was associated with RV GLS (β = .2, p = .03), independent of clinical and echocardiographic variables. RVH+ patients with ApHCM have worse biventricular mechanics and myocardial work, and more heart failure hospitalization, as RVH- at mid-term follow-up.


Angiosarcoma of the left atrium mimicking myxoma: case report.
The case of a 65-year-old female patient with a pedunculated left atrial angiosarcoma is presented. Histologically, the tumor was composed of vascular spaces lined by atypical cells with similar cells in the solid portion of the tumor. Immunohistochemical stain for factor VIII-related antigen was positive in the cells forming vascular spaces. Immunoperoxidase stain for Ulex Europaeus I lectin was positive in cells lining vascular lumina and in individual cells in the solid portion of the tumor. Cardiac angiosarcomas characteristically occur in men, arise in the right atrium, are large mural masses that infiltrate widely at the time of presentation, and are usually rapidly fatal. Review of the literature reveals that this case of cardiac angiosarcoma is unusual by virtue of its occurrence in a female, left atrial location as a polypoid lesion, and prolonged symptom-free survival after surgical resection alone. Cardiac angiosarcoma may clinically and radiologically simulate myxoma.


Follistatin is a developmentally regulated cytokine in neural differentiation.
Activin acts mitogenically on P19 cells as well as being inhibitory of the differentiation of retinoic acid-treated P19 cells and some neuroblastoma cell lines. Here, we show some lines of evidence that follistatin, an activin-binding protein, is also involved in neural differentiation. Counteracting the activity of activin, addition of follistatin suppresses the anchorage-independent growth of P19 cells in soft agar and stimulates neurite outgrowth of a neuroblastoma cell line, IMR-32 cells. While activin does not seem to be expressed significantly, follistatin is demonstrated in the conditioned medium of these cells. Furthermore, the expression of follistatin in P19 cells is subject to dynamic fluctuations in response to retinoic acid treatment. These neural cells may produce follistatin in a cell stage-specific manner in order to interact with exogenously derived activin.


Neoadjuvant chemotherapy in urothelial bladder cancer: impact of regimen and variant histology.
We compared the efficacy of methotrexate/vinblastine/doxorubicin/cisplatin (MVAC) versus gemcitabine/cisplatin in urothelial cancer and neoadjuvant chemotherapy (NACT) efficacy in variant histology (VH). Radical cystectomy patients were retrospectively compared with those who received NACT. Factors associated with survival, pathologic complete response (pCR) and downstaging (pDS) were evaluated in multivariable models. 9% of radical cystectomy patients (84/919) received NACT, with improved survival, pCR and pDS on both regimens. MVAC lead to higher pDS without an increase in pCR. On multivariable analysis, there was a nonsignificant increase in pDS with MVAC. NACT conferred similar responses in squamous and glandular differentiation VH. NACT was associated with improved survival, pCR and pDS. Furthermore, responses to NACT were not dependent on presence of VH.


Comparison of catalytic properties of free and immobilized cellobiase novozym 188.
The enzyme cellobiase from Novo was immobilized in controlled pore silica particles by covalent binding with the silane-glutaraldehyde method with protein and activity yields of 67 and 13.7%, respectively. The activity of the free enzyme (FE) and immobilized enzyme (IE) was determined with 2 g/L of cellobiose, from 40 to 75 degrees C at pH 3.0-7.0 for FE and from 40 to 70 degrees C at pH 2.2-7.0 for IE. At pH 4.8 the maximum specific activity for the FE and IE occurred at 65 degrees C: 17.8 and 2.2 micromol of glucose/(min x mg of protein), respectively. For all temperatures the optimum pH observed for FE was 4.5 whereas for IE it was shifted to 3.5. The energy of activation was 11 kcal/mol for FE and 5 kcal/mol for IE at pH 4.5-5, showing apparent diffusional limitation for the latter. Thermal stability of the FE and IE was determined with 2 g/L of cellobiose (pH 4.8) at temperatures from 40 to 70 degrees C for FE and 40 to 75 degrees C for IE. Free cellobiase maintained its activity practically constant for 240 min at temperatures up to 55 degrees C. The IE has shown higher stability, retaining its activity in the same test up to 60 degrees C. Half-life experimental results for FE were 14.1, 2.1, and 0.17 h at 60, 65, and 70 degrees C, respectively, whereas IE at the same temperatures had half-lives of 245, 21.3, and 2.9 h. The energy of thermal deactivation was 80.6 kcal/mol for the free enzyme and 85.2 kcal/mol for the IE, suggesting stabilization by immobilization.


Sweat testing infants detected by cystic fibrosis newborn screening.
Describe and define limitations of early pilocarpine iontophoresis (sweat testing) for cystic fibrosis (CF) newborn screening (NBS). Population-based results from follow-up of CF NBS-positive newborns. Insufficient quantity of sweat is more likely if the sweat test is done too early, but testing is generally successful after 2 weeks of age. Sweat chloride levels drop over the first weeks of life. CF carriers have higher sweat chloride concentrations than non-carriers. Sweat testing can be performed effectively after 2 weeks of age for CF NBS-positive newborns. Earlier testing has a higher risk of insufficient sweat for completing testing.


The value of splenic preservation with distal pancreatectomy.
Splenic-preserving distal pancreatectomy for benign or low-grade malignant disease is associated with decreased perioperative morbidity compared with conventional distal pancreatectomy with splenectomy. A retrospective review of a prospective database of patients. Memorial Sloan-Kettering Cancer Center, New York, NY. All patients (N = 211) undergoing distal pancreatectomy. Perioperative complications, length of postoperative stay, and overall survival times were analyzed. After excluding patients with adenocarcinoma and those who had other major organ resection, 125 patients underwent distal pancreatectomy for benign or low-grade malignant disease with splenectomy (n = 79) or splenic preservation (n = 46). Perioperative complications occurred in 39 (49%) of the 79 patients following splenectomy and 18 (39%) of the 46 patients following splenic preservation (P =.21). Perioperative infectious complications and severe complications were significantly higher in the splenectomy group (28% and 11%) compared with the splenic preservation group (9% and 2%) (P =.01 and.05), respectively. Length of hospital stay was 9 days (range, 5-41 days) following splenectomy and 7 days (range, 5-26 days) following splenic preservation (P<.01). No difference in length of surgery, units of blood transfused, or perioperative mortality was noted between groups. Splenic preservation following distal pancreatectomy for benign or low-grade malignant disease is safe and is associated with a reduction in perioperative infectious complications, severe complications, and length of hospital stay compared with conventional distal pancreatectomy with splenectomy. Therefore, splenic preservation should be considered in this group of patients.


Presence of cerebral amyloid modulates phenotype and pattern of neurodegeneration in early Parkinson's disease.
To evaluate the frequency of cerebral amyloid in early Parkinson's disease (ePD) and provide a multimodal assessment of the influence of cerebral amyloid on disease phenotype. We performed a multicentre cohort study of the Parkinson's Progression Markers Initiative (PPMI), including 369 drug-naïve patients with ePD and 174 healthy controls (HC). Cerebrospinal fluid (CSF) amyloid-β levels were transformed using the linear regression procedure. A cut-off of >198 pg/mL was used to define amyloid-negative (PD-) and amyloid-positive (PD+) subgroups. Grey matter (GM) density and hippocampal volume from the MRI was measured using Advanced Normalisation Tools (ANTs). We compared demographic, genetic, CSF, behavioural, functional and imaging modalities across ePD- and ePD+ groups. We observed that 16.5% of ePD have CSF evidence of amyloidosis. PD+ was significantly older than PD-, had a higher frequency of APOE e4 alleles and all CSF measures (total-tau, phosphorylated-tau and α-synuclein) were reduced. PD+ had reduced cognitive performance relative to PD- on Symbol-Digit Matching, Verbal Category Fluency and Delayed Recall tests. Imaging analysis in a subset of individuals (PD+ =43; PD- =241) revealed overlapping GM atrophy relative to HC in medial temporal, frontal and brainstem structures. Direct comparisons revealed PD+ GM reductions predominantly located in the frontal cortex while PD- had GM reductions in subcortical structures. These observations remain when controlling for age and APOE e4 allele status. Cerebral amyloid in ePD yields a unique phenotype across all measured modalities that is consistent with a synergistic interaction between α-synuclein and amyloid pathology. Amyloid status should be considered when screening these individuals for trials involving disease-modifying agents.


Gene concepts and genethics: beyond exceptionalism.
The discursive explosion that was provoked by the new genetics could support the impression that the ethical and social problems posed by the new genetics are somehow exceptional in their very nature. According to this view we are faced with special ethical and social problems that create a challenge so fundamental that the special label of genethics is needless to justify. The historical account regarding the evolution of the gene concepts could serve us to highlight the limits of what we know about genes and what we can do with genes. The widespread notion about the exceptionality of genetic knowledge and its applicative possibilities is hardly justifiable and leads to misunderstandings regarding the conceptualization of the ethical and social problems we might face. Following a more realistic interpretation of the role of genes in human life we might avoid a whole set of fictive dilemmas and counterproductive regulatory efforts in bioethics. Bioethical discourse should move from the gene-centered scientific discourse toward the more sophisticated and complex discourses where human development represented as a matter of complex interactions between genomes and environments, between genes, educational factors, nutritional regimes, and other different developmental resources. If a gene is seen as one among the different developmental resources that are shaping a given human trait then both genethics and genetic exceptionalism could hardly be represented as a justified approach in discussing the ethical and social problems of genetics.


The association between depression and suicide when hopelessness is controlled for.
We retested the relationship between major depression and suicide with hopelessness as a control variable, with the hypothesis that the strong relationship between depression and suicide will decrease or disappear when hopelessness is controlled for. Also, hopelessness can be accounted for by psychological strains that resulted from social structure coupled with individual characteristics. This was a case-control psychological autopsy study, in which face-to-face interviews were conducted to collect information from proxy informants for suicide victims and living subjects in rural Chinese 15-34years of age who died of suicide (n=392) and who served as community living controls (n=416). Major depression was assessed by the Chinese version of the Structured Clinical Interview for DSM-IV (SCID). Hopelessness was measured by Beck Hopelessness Scale. A strong association between major depression and suicide was observed after adjustment for socio-demographic characteristics. When hopelessness was added to the analysis, the depression-suicide relationship was significantly decreased in all the six regression models. Although depression, as well as other mental illness, is a strong risk factor for suicide, depression and suicide are both likely to be related to hopelessness, which in turn could be a consequence of psychological strains that resulted from social structure and life events. Future studies may examine the causal relations between psychological strains and hopelessness.


Effects of echinacea on the frequency of upper respiratory tract symptoms: a randomized, double-blind, placebo-controlled trial.
Upper respiratory tract infection symptoms are a common cause of morbidity. Herbal preparations of the plant Echinacea purpurea have immune-enhancing properties. To compare the frequency of upper respiratory tract symptoms in individuals receiving E. purpurea capsules and those receiving placebo to evaluate the preventive efficacy of echinacea. In a randomized, double-blind clinical trial, 90 volunteers recruited from hospital personnel were randomly assigned to receive 3 capsules twice daily of either placebo (parsley) or E. purpurea for 8 weeks during the winter months. Upper respiratory tract symptoms were reported weekly during this period. Fifty-eight individuals were included in the final data analysis: 28 in the echinacea group and 30 in the placebo group. Individuals in the echinacea group reported 9 sick days per person during the 8-week period, whereas the placebo group reported 14 sick days (z = -0.42; P = .67). Mild adverse effects were noted by 8% of the echinacea group and 7% of the placebo group (P = .24). Prophylactic treatment with commercially available E. purpurea capsules did not significantly alter the frequency of upper respiratory tract symptoms compared with placebo use.


Inflammatory bowel disease patients provide reliable self-reported medical information: A multicentre prospective pharmacovigilance monitoring system.
To assess the agreement between patient-reported and health care provider-reported medical information in inflammatory bowel disease (IBD). This multicentre, prospective, event monitoring study enrolled adult Crohn's disease (CD) and ulcerative colitis (UC) patients treated with a biological in four medical centers in the Netherlands. At two-monthly intervals, patients completed questionnaires on biological use, combination therapy and indication. The patient-reported information was compared with their electronic health records (EHRs) and analysed for percentage agreement and Cohen's kappa. A reference population from a prospective IBD registry was used to assess the representativeness of the study population. In total, 182 patients (female 50.5%, mean age 42.2 years, CD 76.9%) were included in the analysis. At baseline, 51.0% of the patients were prescribed an immunomodulator (43.9% thiopurines, 7.1% methotrexate), and patients were prescribed biologicals as follows: 59.3% infliximab, 30.2% adalimumab, 9.3% vedolizumab, and 1.1% ustekinumab. Agreement on patient-reported indication and biological use was almost perfect (κ = 0.878 and κ = 1.000, respectively); substantial for combination therapy (κ = 0.672). Gender, age, type of IBD, biological use and combination therapy were comparable with the reference population. Systematic patient-reporting by questionnaires was reliable in retrieving indication and treatment specific information from IBD patients. These results indicate that the use of patient-reporting outcomes in daily IBD practice can ensure reliable information collection.


Loss of Hepatic Transcription Factor EB Attenuates Alcohol-Associated Liver Carcinogenesis.
Alcohol is a well-known risk factor for hepatocellular carcinoma. Autophagy plays a dual role in liver cancer, as it suppresses tumor initiation and promotes tumor progression. Transcription factor EB (TFEB) is a master regulator of lysosomal biogenesis and autophagy, which is impaired in alcohol-related liver disease. However, the role of TFEB in alcohol-associated liver carcinogenesis is unknown. Liver-specific Tfeb knockout (KO) mice and their matched wild-type (WT) littermates were injected with the carcinogen diethylnitrosamine (DEN), followed by chronic ethanol feeding. The numbers of both total and larger tumors increased significantly in DEN-treated mice fed ethanol diet than in mice fed control diet. Although the number of tumors was not different between WT and L-Tfeb KO mice fed either control or ethanol diet, the number of larger tumors was less in L-Tfeb KO mice than in WT mice. No differences were observed in liver injury, steatosis, inflammation, ductular reaction, fibrosis, and tumor cell proliferation in DEN-treated mice fed ethanol. However, the levels of glypican 3, a marker of malignant hepatocellular carcinoma, markedly decreased in DEN-treated L-Tfeb KO mice fed ethanol in comparison to the WT mice. These findings indicate that chronic ethanol feeding promotes DEN-initiated liver tumor development, which is attenuated by genetic deletion of hepatic TFEB.


The effect of mannitol upon cochlear dysfunction induced by transient local anoxia.
Transient local anoxia of the cochlea was induced by pressing the labyrinthine artery, and compound action potential (CAP) or endocochlear potential (EP) was measured before and after transient local anoxia ranging from 5 to 60 min using 106 albino guinea pigs. The complete interruption of the cochlear blood flow by this procedure and its full restoration after releasing the pressure on the artery was confirmed by a laser-Doppler flowmeter. The anoxia of less than 10 min induced no post-anoxic cochlear dysfunction, whereas the anoxia of a longer duration induced an irreversible dysfunction of the cochlea. It was evident that the post-anoxic recovery of the CAP threshold was worse as the anoxia period was prolonged, and CAP was almost completely abolished after 60-min anoxia. In animals which were administered mannitol intravenously just after the restoration of the cochlear blood circulation, the recovery of the CAP threshold was significantly better than that in the control animals, when the animals were subjected to local anoxia of 15- to 30-min duration. No beneficial effect, however, was observed in the 60-min anoxia group. In conclusion, local anoxia of 10 min or longer caused cochlear dysfunction, which was partially but significantly alleviated by mannitol.


Sleep symptoms and long-term outcome in adolescents with major depressive disorder: a naturalistic follow-up study.
Sleep abnormalities in major depressive disorder (MDD) have been suggested to represent a vulnerability trait, which might predispose the individual to long-term psychiatric morbidity. In this study, we sought to assess whether the presence of sleep symptoms among adolescents with MDD is associated with poorer long-term outcome in young adulthood during naturalistic follow-up. Adolescent outpatients diagnosed with MDD (n = 166; age 13-19 years, 17.5% boys) were followed up during 8 years in naturalistic settings. N = 112 adolescents (16.1% boys) completed the 8-year assessment. Sleep symptoms and psychosocial functioning were assessed with structured clinical interviews, and depressive and anxiety symptoms with questionnaires. The severity of sleep symptoms at baseline was not associated with worse outcome at 8 years in terms of any of the outcome measures tested. In particular, the presence of a disturbed sleep-wake rhythm at baseline was associated with a more favourable outcome at 8 years: less depression and anxiety symptoms and higher level of psychosocial functioning. The presence of sleep symptoms in young adulthood was associated with the presence of current depression and anxiety symptoms and poorer psychosocial functioning. The presence of sleep symptoms at follow-up seems to be state-dependent: they are observed in conjunction with other psychiatric symptoms. Contrary to our hypothesis, our results suggest that sleep complaints among adolescents with MDD do not lead to poorer long-term clinical outcome in young adulthood. The link between sleep-wake rhythm disturbance and better long-term outcome needs to be confirmed and examined in detail in further studies, but here we speculate about possible explanations.


Rhabdomyolysis after laparoscopic bariatric surgery.
Postoperative rhabdomyolysis is an uncommon event. The aim of this study was to determine the incidence of rhabdomyolysis following laparoscopic obesity surgery. Rhabdomyolysis was studied prospectively. Over a 6-month period, 66 consecutive patients underwent bariatric surgery (gastric banding (n=50) and gastric bypass (n=16)). All patients underwent laparoscopic procedures. A range of blood tests, including serum creatine phosphokinase (CPK) level and serum creatinine, were systematically performed before surgery, and on the first and third day postoperatively. Rhabdomyolysis was defined as a postoperative CPK level >1050 IU/L. Serum CPK was noted to increase significantly postoperatively to >1050 units in 3 patients (6%) in the adjustable banding group and 12 patients (75%) in the gastric bypass group (P <0.01). In the bypass group, 4 patients (25%) had a serum CPK level >10000 IU/L, but there were none in the gastric banding group. All patients with CPK level >10000 IU/L had BMI >60 kg/m(2). No patients experienced acute renal failure. Rhabdomyolysis occurred in 22.7 % of 66 consecutive patients undergoing laparoscopic bariatric surgery. Risk factors were identified: massive obesity and long duration of the operation. Early diagnosis may have significant impact on outcome by preventing or reducing the severity of complications from rhabdomyolysis. CPK level should be performed systematically after obesity surgery.


Interaction of the yeast pleiotropic drug resistance genes PDR1 and PDR5.
The network of genes which mediates multiple drug resistance in yeast includes, among others, the PDR1 gene, which encodes a putative regulator of gene expression, and PDR5, a locus whose amplification leads to resistance. We demonstrate that disruption of PDR5 causes marked hypersensitivity not only to cycloheximide but also to sulphometuron methyl and the mitochondrial inhibitors chloramphenicol, lincomycin, erythromycin and antimycin. Genetic analysis of double mutants containing an insertion in PDR5 (pdr5:Tn5), which renders cells hypersensitive to cycloheximide, and a pdr1 mutation, which confers resistance to this inhibitor, indicates that the expression of resistance requires a functional PDR5 gene. The same interdependency is observed for chloramphenicol, but not for oligomycin, lincomycin, erythromycin or sulphometuron methyl. Northern analysis of PDR1 and PDR5 transcripts reveals that the 5.2 kbp PDR5 transcript is overexpressed in pdr1 (resistant) mutants, but underexpressed in a disruption of PDR1. These observations provide strong experimental support for our former proposal that the PDR5 gene is a target for regulation by the PDR1 gene product.


Achromobacter xylosoxidans mesh related infection: a case of delayed diagnosis and management.
We present the first case of mesh related infection caused by Achromobacter xylosoxidans after ventral hernia repair. After repair of a small paraumbilical hernia, the postoperative course was complicated by persistent discharging sinuses despite the removal of underlying polypropylene mesh. Removal of an intrabdominal omental inflammatory mass containing pus that showed growth of A. xylosoxidans led to the resolution of all the symptoms.


[Biventricular stimulation for AV block].
Since the establishment of cardiac resynchronization therapy in left bundle branch block and mechanical asynchrony, the adverse effects of right ventricular apical pacing have gained increasing scientific interest. The sequelae of "iatrogenic desynchronization" on cardiac structure and function as well as on patients' prognosis could be well documented. "Minimally desynchronizing" stimulation strategies could be an alternative for patients needing ventricular pacing. The search for hemodynamically more advantageous alternative right ventricular pacing sites has failed so far to deliver well validated results, and due to the somewhat higher lead dislodgment rates pure left ventricular pacing cannot be recommended, at least not in pacemaker-dependent patients. Hence there is the question for primary biventricular stimulation in patients with AV block. The results of several biventricular studies with limited numbers of patients have been promising with respect to structural and functional surrogate endpoints. Two major controlled prospective and prognostically orientated studies, the BIOPACE study and the BLOCK-HF study, are currently underway and will report results in the next few years. According to the actual guidelines of the European Society of Cardiology (ESC) the implantation of a biventricular system is recommended in patients with AV block even without left bundle branch block (Class IIa, evidence level C) if they fulfill the remaining criteria that justify the implantation of a biventricular system. According to the guidelines for pacemaker therapy of the German Cardiac Society (GCS) biventricular pacing can be considered in these patients. Both societies do expressly permit the implantation of biventricular systems with ICD backup if indicated.


Bacterial meningitis caused by nontraumatic cerebrospinal fluid rhinorrhea with aqueductal stenosis: A case report.
Nontraumatic cerebrospinal fluid (CSF) rhinorrhea associated with aqueductal stenosis is rare. The resulting CSF leakage may cause bacterial meningitis, and appropriately timed surgical treatment should be considered. A 28-year-old woman with obstructive hydrocephalus secondary to aqueductal stenosis presented with intermittent nasal discharge. CSF rhinorrhea was suspected, but she refused surgery. During the course of conservative treatment, she developed meningitis. Exacerbation of hydrocephalus and CSF rhinorrhea was suspected, and the patient underwent endoscopic third ventriculostomy after recovery from meningitis. Postoperatively, ventricular size decreased and CSF leakage completely resolved. There was no recurrence of hydrocephalus or rhinorrhea. Patients with intermittent CSF rhinorrhea due to exacerbation of hydrocephalus are at high risk for bacterial meningitis. Appropriately timed surgical treatment results in a favorable outcome.


Electron trajectories associated with laser-driven coherent synchrotron emission at the front surface of overdense plasmas.
We present an in-depth analysis of an ultrafast electron trajectory type that produces attosecond electromagnetic pulses in both the reflected and forward directions during normal incidence, relativistic laser-plasma interactions. Our particle-in-cell simulation results show that for a target which is opaque to the frequency of the driving laser pulse the emission trajectory is synchrotronlike but differs significantly from the previously identified figure-eight type which produces bright attosecond bursts exclusively in the reflected direction. The origin and characteristics of this trajectory type are explained in terms of the driving electromagnetic fields, the opacity of the plasma, and the conservation of canonical momentum.


Free radicals. Formation, function and potential relevance in anaesthesia.
Free radical species are ubiquitous in plant and animal life. This article describes briefly the formation of certain oxygen-centred free radicals which are essential for aerobic metabolism and host defences in humans. The mechanism of cytotoxicity of excess or inappropriate free radical production is described. The potential relevance of free radical tissue injury to the anaesthetist is illustrated using oxygen toxicity, adult respiratory distress syndrome and halothane hepatitis as examples.


Identification and significance of sulphonamides (p-TSA, o-TSA, BSA) in an urban water cycle (Berlin, Germany).
Because of the nature of the water cycle in Berlin, a number of persistent wastewater residues are present in the surface water and may potentially reach the groundwater via bank filtration and artificial recharge. The occurrence and behaviour of the wastewater residues para-toluenesulphonamide (p-TSA), ortho-toluenesulphonamide (o-TSA) and benzenesulphonamide (BSA) through wastewater treatment, surface water, bank filtration and drinking water treatment was studied. In addition, groundwater below a former sewage farm was investigated. All three compounds are ubiquitous in the aquatic environment of Berlin. p-TSA concentrations are much higher than those of o-TSA and BSA. p-TSA was found in high concentrations in the wastewater influent (2 to 15microg/L), in the wastewater effluent (<0.15 to 2.34microg/L) and in sewage farm groundwater (<0.05 to 20microg/L) and in lower concentrations in the surface water (<0.05 to 1.15microg/L), bank filtrate (<0.05 to 0.30microg/L) and drinking water (<0.05 to 0.54microg/L). p-TSA is considerably depleted during waste- and drinking water treatment ( approximately 90% reduction each). The concentration ranges for o-TSA and BSA in wastewater influents were 0.11 to 8microg/L and <0.05 to 0.64microg/L, respectively, while the values for wastewater effluents were 0.14 to 4microg/L for o-TSA and 0.25 to 0.49microg/L for BSA. Wastewater treatment and drinking water treatment do not reduce the concentrations of o-TSA and BSA. The behaviour of o-TSA during wastewater treatment varies largely between different wastewater treatment plants where concentrations increase, remain constant or decrease. BSA forms during treatment. The concentrations measured in surface water, sewage farm groundwater, bank filtrate and drinking water were <0.05 to 1.74microg/L for o-TSA and <0.05 to 0.53microg/L for BSA.


Pathogenesis and treatment of pancreatitis due to essential fatty acid deficiency.
Pathogenesis of pancreatitis was studied in experimental animals and essential fatty acid deficiency was condemned as an etiological factor, since it caused pathological changes in permeability of cell membrane of the pancreatic acinar cells. Favorable effects of intravenous fat emulsion was clearly demonstrated in treatment for acute and chronic pancreatitis.


A neglected posterior fracture dislocation of the shoulder with ipsilateral distal humeral shaft fracture.
Cases of posterior dislocation or fracture-dislocation of the shoulder are rare. A combination of posterior dislocation and ipsilateral humeral shaft fracture, especially a distal third one, is even rarer. In such injuries, the diagnosis of shoulder dislocation may be missed because great attention is usually given to the fractured humerus. We present this case report to raise awareness among emergency physicians of this manner of injury. We present a case report of a 30-year-old woman who had posterior fracture dislocation of the right shoulder with ipsilateral distal humeral shaft fracture. The shaft fracture was initially evaluated, internally reduced, and fixed, whereas the posterior fracture-dislocation of the shoulder was not recognized. A belated diagnosis was established 1 month after the operation, when a computed tomography scan was performed because the patient reported continuous pain and limited range of motion of the shoulder. A second reduction was performed to reduce the dislocation. The patient had partial functional recovery 1 year later. Consideration of this condition, coupled with thorough and appropriate physical and radiological examinations, could lead to improved recognition of such cases.


Echocardiographic assessment of pulmonary arterial pressure in children with complete right bundle branch block.
The time interval between tricuspid valve closure and pulmonary valve opening, termed the isovolumic contraction time of the right ventricle, was evaluated echographically in 38 normal children and within 24 hours of cardiac catheterization in 53 children with congenital heart disease and normal conduction as assessed with the electrocardiogram. In the 53 patients with congenital heart disease, isovolumic contraction time was strongly influenced by right ventricular afterload, as defined by pulmonary arterial end-diastolic pressure (r = 0.87). It was possible to utilize isovolumic contraction time to separate patients with normal or elevated values for pulmonary arterial end-diastolic pressure. Similar correlations were demonstrated between isovolumic contraction time and mean pulmonary arterial pressure and calculated pulmonary vascular resistance. Evaluation of 15 children with complete right bundle branch block revealed values for isovolumic contraction time that did not significantly differ from those of patients with similar pulmonary arterial end-diastolic pressure but no conduction abnormalities. These findings indicate that serial echographic evaluation of the interval from tricuspid valve closure to pulmonary valve opening can give an accurate reproducible assessment of right ventricular afterload in many children with congenital heart disease and complete right bundle branch block


Documentation of paediatric head injuries in a mixed metropolitan emergency department.
Head injuries are a common presentation of children to Australian EDs. Healthcare documentation is an important tool for enhancing patient care. In our study, we aimed to assess the adequacy of paediatric head injury documentation in a mixed ED. A retrospective analysis of presentations to a mixed ED between 2017 and 2018. Children aged <16 years old with a primary diagnosis of head injury were included. Documentation items based on local head injury guidelines were assessed in both medical and nursing documentation. We compared cases aged <1 and ≥1 year. There were 427 presentations that met the case definition. Medical documentation was present in 422 cases and nursing documentation in 310 cases. In combined medical and nursing documentation, items poorly documented include blood pressure (BP; 21.3%) and secondary survey (16.9%). In solely medical documentation, least commonly documented items are high-risk bony injuries (22.5%), high-risk soft tissue injuries (22.3%), seizure (22.0%) and non-accidental injury (3.6%). Glasgow Coma Scale (GCS) was poorly documented in cases aged <1 year (10.9%, P < 0.001). The largest gaps in the documentation of paediatric head injuries were BP and paediatric GCS in infants. Future audits and educational strategies should focus on targeting clinically relevant items that are predictive of serious outcomes.


Expression of the metalloproteases MMP-1, MMP-2, MMP-3, MMP-9, MMP-11, TIMP-1 and TIMP-2 in angiocentric midfacial lymphomas.
Extranodal T/NK cell lymphomas possess distinctive clinico-pathological characteristics: they are angiocentric, exhibit extensive necrosis. Prognosis is poor in the short term. The objective is to explore the expression of different MMPs in the cells and stroma which are around of the blood vessels damaged and their correlation with clinico-pathological parameters. Twenty cases of this type of lymphomas were studied and collected patient clinical data. The expressions of MMP-1, 2, 3, 9, 11, 13 and TIMP-1, 2 were studied by immunohistochemistry. Ultrastructural studies were performed in two cases. Statistical analysis was done with Fisher's exact test, Chi(2) test. Of the 20 patients, 13 were men with median age of 43 years. In 13 patients the primary tumor was localized in the nasal cavity. Treatment was combined chemotherapy and radiotherapy in 60%. The 55% advanced clinical stages, 70% died from the disease. There were neoplastic cell and peritumoral fibroblasts positivity to MMP-1 and MMP-11 in most of the cases. The MMPs-2, 3 and 9 were expressed in neoplastic cell between 30 to 65%of the cases. TIMP-1 was presented mainly in the epithelium and TIMP-2 was poor expressed of the all cases. There were no statistical significance between the different enzymes used and the clinical parameters, besides status and survival of the patients. It is necessary to study more enzymes and focus them to quantify and determine their activity, in order to have a better correlation with histological features in this type of neoplasm.


Relationship between nailfold capillary abnormalities and vestibular dysfunction in systemic sclerosis.
To investigate the vestibular dysfunction in a cohort of patients with systemic sclerosis (SSc) and to correlate the findings with disease parameters and microvascular involvement. Vestibular affection was assessed in 30 female SSc patients and 29 age-matched healthy females subjects by using the computerized dynamic platform posturography (CDP). Assessment of microvasculature was done by nailfold videocapillaroscopy (NVC). The main clinical correlates of disease, such as renal function, skin, articular and lung involvement, were evaluated by clinical and instrumental investigations. Subtle vestibular dysfunction was detected in 33% of SSc patients. They exhibited significant decrease in their vestibular ratio values compared to controls (P=0.01). There was a statistical significant association of vestibular affection with both Rodnan's skin score and vascular severity score. Moreover a significant association was found between vestibular dysfunction and NVC patterns. On the other hand, no correlation was observed between vestibular impairment with age, disease duration, disease subsets, autoantibodies and the other clinical disease parameters. Our results showed an evidence of vestibular impairment in patients with SSc. Vestibular dysfunction positively correlates with vascular severity score as assessed by NVC.


Phenomenon of declining anxiety sensitivity scores: a controlled investigation.
Repeated administration of anxiety sensitivity measures can often produce declining scores, even in ostensible control groups, which is a significant concern for researchers. The reasons for these changes are as yet unknown, but could be because of regression to the mean in samples selected on extreme scores, exposure to general information about anxiety contained in psychiatric interviews, or mere exposure to anxiety sensitivity information. This study sought to experimentally evaluate these potential explanations using a comprehensive measure of anxiety sensitivity and its subcomponents, a non-anxiety sensitivity measure (self-esteem), and participants representing the full spectrum of anxiety sensitivity. Results indicated significant decreases in anxiety sensitivity scores (but not self-esteem scores) that could not be accounted for by regression to the mean or exposure to information about anxiety in general. Several potential explanations for these findings are reviewed and implications for research study designs are discussed.


Validation of a method for the determination of sterols and triterpenes in the aerial part of Justicia anselliana (Nees) T. Anders by capillary gas chromatography.
An accurate and sensitive method, combining soxhlet extraction, solid phase-extraction and capillary gas chromatography is described for the quantitative determination of one triterpene (lupeol) and three sterols (stigmasterol, campesterol and beta-sitosterol) and the detection of another triterpene (alpha-amyrin) from the aerial part of Justicia anselliana. This is the first method allowing the quantification of sterols and triterpenes in this plant. It has been fully validated in order to be able to compare the sterol and triterpene composition of different samples of J. anselliana and therefore help to explain the allelopathic activity due to these compounds. This method showed that the aerial part of J. anselliana contained (292+/-2)mg/kg of lupeol, (206+/-1)mg/kg of stigmasterol, (266+/-2)mg/kg of campesterol and (184+/-9)mg/kg of beta-sitosterol.


An ion exchange chromatography and radioimmunoassay procedure for measuring opioid peptides and substance P.
The measurements of peptides of the enkephalin, dynorphin and substance P systems is complicated by the number of possible precursor fragments and degradation products that might cross-react with the antisera. By using an ion-exchanger step before radioimmunoassay we can reduce the possibility that observed peptide levels are due to precursors or metabolites. The ion-exchanger method runs with good recovery and its main advantage is that many samples can be run in parallel. The recovery from the ion-exchanger was similar using two different homogenizing media, whereas the measured endogenous levels of [Met] and [Leu]enkephalin were 3-4 fold higher with 1M acetic acid than when a 1:1 MeOH/HCl mixture was used for tissue extraction.


Achieving the composite endpoint of glycated haemoglobin <7.0%, no weight gain and no hypoglycaemia in the once-weekly dulaglutide AWARD programme.
To compare the effectiveness of dulaglutide 1.5 and 0.75 mg with active comparators and placebo with regard to a composite endpoint of glycated haemoglobin (HbA1c), weight and hypoglycaemia, using post hoc analyses. A logistic regression analysis was performed on the intention-to-treat population, using data from the last observation carried forward, and the composite endpoint of HbA1c <7.0% (53 mmol/mol), no weight gain (≤0 kg) and no hypoglycaemia (glucose <3.0 mmol/l or severe hypoglycaemia) after 26 weeks for each trial in the AWARD programme separately. At 26 weeks, within each study, 37-58% of patients on dulaglutide 1.5 mg, 27-49% of patients on dulaglutide 0.75 mg, and 9-61% of patients on active comparators achieved the composite endpoint. Significantly more patients reached the composite endpoint with dulaglutide 1.5 mg than with metformin, sitagliptin, exenatide twice daily or insulin glargine: odds ratio (OR) 1.5 [95% confidence interval (CI) 1.0, 2.2; p < 0.05], OR 4.5 (95% CI 3.0, 6.6; p < 0.001), OR 2.6 (95% CI 1.8, 3.7; p < 0.001) and OR 7.4 (95% CI 4.4, 12.6; p < 0.001), respectively, with no difference between dulaglutide 1.5 mg and liraglutide 1.8 mg. In addition, significantly more patients reached the composite endpoint with dulaglutide 0.75 mg than with sitagliptin or insulin glargine: OR 3.3 (95% CI 2.2, 4.8; p < 0.001) and OR 4.5 (95% CI 2.7, 7.8; p < 0.001), respectively. Dulaglutide is an effective treatment option, resulting in a similar or greater proportion of patients reaching the HbA1c target of <7.0% (53 mmol/mol), without weight gain or hypoglycaemia compared with active comparators.


Phase I-II study of irinotecan combined with mitomycin-C in patients with advanced gastric cancer.
Irinotecan (CPT-11) shows synergism with mitomycin-C (MMC) in a preclinical setting. The goals of this study were to determine the maximum tolerated dose (MTD), the dose limiting toxicity, the recommended dose (RD), and preliminary anti-tumor activity in a combined CPT-11 and MMC treatment of advanced gastric cancer. The study was designed to evaluate escalated doses of CPT-11 and MMC administered every two weeks. Five escalating dose levels were studied (CPT-11/ MMC: 100/5; 125/5; 150/5; 150/7; 150/10 mg/m2). Thirty-one patients were enrolled. Thirty patients were assessable for toxicity and tumor response for 89 treatment cycles. The median age was 60 years (32-73 years), and most patients (90%) had a performance status of 0 to 1. Fourteen patients were previously treated and 17 were chemotherapynaive. The MTD was CPT-11 150 mg/m2 plus MMC 10 mg/m2, in which all three patients experienced grade 4 neutropenia. including one episode of prolonged and one of febrile neutropenia, and one patient experienced grade 3 diarrhea during the first cycle. Fifteen partial responses were observed. The RD based on this phase I-II study was CPT-11 150 mg/m2 plus MMC 5 mg/m2 administered every two weeks. This combination demonstrates promising activity against advanced gastric cancer and warrants further investigation in another phase II study.


Hematoma in a low-grade medullary astrocytoma: report of an unusual case and literature review.
We describe the case - to our knowledge unique - of an 8-year-old boy who presented with acute onset of lower cranial nerve palsy and tetraparesis caused by a hematoma in a dorsal exophytic pilocytic astrocytoma of the medulla oblongata. The boy showed near-complete recovery after neurosurgical management in two stages: first, emergency evacuation of the hematoma with tumor biopsy, and second, complete tumor removal 5 months after the initial event. Intraoperative electrophysiological monitoring techniques for the lower cranial nerves are of value in preserving their functional integrity. Ultrasonography is helpful in assessing the extent of tumor removal. Although the pathological diagnosis of a pilocytic astrocytoma would not justify radiotherapy, local field radiotherapy was added mainly because of the unexpectedly rapid tumor progression during the interval between the two surgical procedures. The literature on brainstem and tumor hematoma in children is reviewed.