Translating neurobehavioural endpoints of developmental neurotoxicity tests into in vitro assays and readouts.
The developing nervous system is particularly vulnerable to chemical insults. Exposure to chemicals can result in neurobehavioural alterations, and these have been used as sensitive readouts to assess neurotoxicity in animals and man. Deconstructing neurobehaviour into relevant cellular and molecular components may allow for detection of specific neurotoxic effects in cell-based systems, which in turn may allow an easier examination of neurotoxic pathways and modes of actions and eventually inform the regulatory assessment of chemicals with potential developmental neurotoxicity. Here, current developments towards these goals are reviewed. Imaging genetics (CB) provides new insights into the neurobiological correlates of cognitive function that are being used to delineate neurotoxic mechanisms. The gaps between in vivo neurobehaviour and real-time in vitro measurements of neuronal function are being bridged by ex vivo measurements of synaptic plasticity (RW). An example of solvent neurotoxicity demonstrates how an in vivo neurological defect can be linked via the N-methyl-d-aspartate (NMDA)-glutamate receptor as a common target to in vitro readouts (AB). Axonal and dendritic morphology in vitro proved to be good correlates of neuronal connectivity and neurobehaviour in animals exposed to polychlorinated biphenyls and organophosphorus pesticides (PJL). Similarly, chemically induced changes in neuronal morphology affected the formation of neuronal networks on structured surfaces. Such network formation may become an important readout for developmental neurotoxicity in vitro (CvT), especially when combined with human neurons derived from embryonic stem cells (ML). We envision that future in vitro test systems for developmental neurotoxicity will combine the above approaches with exposure information, and we suggest a strategy for test system development and cell-based risk assessment.


Evidence that a single gene with gender- and age-dependent effects influences systolic blood pressure determination in a population-based sample.
A biometrical study was carried out to evaluate the role of genetic variation in determining interindividual differences in systolic blood pressure (SBP) in the population at large. SBP was measured in 1,266 Caucasian individuals in 278 pedigrees ascertained through children enrolled in the Rochester, MN, school system. The sample included 646 males and 620 females 550 years of age and not taking antihypertensive medication or oral contraceptives. Complex segregation analysis was first applied to these data by using a regression model for age, in which the intercept was gender and ousiotype specific but in which the slope was only gender specific. When the slope was independent of ousiotype, neither variation at a single gene combined with polygenic effects (mixed genetic model) nor variation in a single environmental factor combined with polygenetic effects (mixed environmental model) explained the distribution of SBP in this sample. However, when the regression model for age allowed both the intercept and slope to be gender and ousiotype specific, the mixed environmental model was rejected whereas the mixed genetic model was not. These results suggest that variability in SBP may be influenced by major effects of allelic variation at a single gene that are both gender and age dependent. This study (1) suggests that particular genotypes determined by a single gene are associated with a steeper increase of SBP with age among males and females 550 years of age in the general population and (2) illustrates the need to consider models that more realistically represent the relationship between genotypic variability and phenotypic variability, to understand the genetics of human quantitative traits.


Molecular characterization of breast cancer in young Brazilian women.
To evaluate the distribution of molecular subtypes of breast tumors diagnosed in young Brazilian women and to analyze the frequency of loss of heterozygocity (LOH) in BRCA1 among different molecular subtypes of early-onset breast cancer. Samples from 72 cases of invasive breast carcinoma diagnosed in women aged between 19 and 40 years were evaluated using an immunohistochemical panel of biomarkers. Three intragenic BRCA1 locus microsatellites, D17S1322, D17S1323, and D17S855, were PCR amplified from matched normal (lymphocyte) and tumor DNAs for (LOH) analysis. We found 13 cases (18%) that had an immunohistochemical profile consistent with being basal-like. Forty cases (55%) were luminal A type; 11% (8 cases) were luminal B type, 13% (9 cases) were HER2-overexpressing tumors and two cases were ER-/HER2- carcinomas lacking basal marker expression. Four of the 16 informative cases at D17S1322, one of the four informative cases at D17S855, and none of the five informative cases at D17S1323 displayed LOH (four basal-like and one Luminal A). Microsatellite instability (MSI) at D17S855 and D17S1322 was found in two cases (one a basal-like and one Luminal A). In our study, basal-like tumor was the second most frequent molecular type among young Brazilian women and was only observed in women diagnosed under the age of 35 years. There was no significant difference of LOH at BRCA1 locus rates between basal-like breast tumors and not-basal-like breast tumors (p=0.62). LOH in BRCA1 and MSI in these breast cancers were not frequent but may indicate a small group of breast cancers with a specific molecular makeup.


Presence of Vaccine-Derived Newcastle Disease Viruses in Wild Birds.
Our study demonstrates the repeated isolation of vaccine-derived Newcastle disease viruses from different species of wild birds across four continents from 1997 through 2014. The data indicate that at least 17 species from ten avian orders occupying different habitats excrete vaccine-derived Newcastle disease viruses. The most frequently reported isolates were detected among individuals in the order Columbiformes (n = 23), followed in frequency by the order Anseriformes (n = 13). Samples were isolated from both free-ranging (n = 47) and wild birds kept in captivity (n = 7). The number of recovered vaccine-derived viruses corresponded with the most widely utilized vaccines, LaSota (n = 28) and Hitchner B1 (n = 19). Other detected vaccine-derived viruses resembled the PHY-LMV2 and V4 vaccines, with five and two cases, respectively. These results and the ubiquitous and synanthropic nature of wild pigeons highlight their potential role as indicator species for the presence of Newcastle disease virus of low virulence in the environment. The reverse spillover of live agents from domestic animals to wildlife as a result of the expansion of livestock industries employing massive amounts of live virus vaccines represent an underappreciated and poorly studied effect of human activity on wildlife.


[Canal-otolithic interaction under conditions of otolithic asymmetry].
Intact pigeons were exposed to horizontal rotations so that the horizontal canal afferents were activated either separately or simultaneously with macular afferents. When the pigeons, head was off the axis of rotation, the radial acceleration (0.5 g) acted either in sagittal or in frontal plane. In presence of otolith asymmetry, the radial acceleration modified the rotation-induced nystagmus depending on the direction: the duration of the rightward nystagmus was increased whereas the duration of the leftward nystagmus was decreased, and vice versa. These changes occurred irrespective of the source of the otolith asymmetries. Theoretical significance of this data is discussed and possible mechanisms of the canal-otolith interaction are considered.


Anti-cataract activity of Pterocarpus marsupium bark and Trigonella foenum-graecum seeds extract in alloxan diabetic rats.
Long-term complications are frequently encountered in diabetes mellitus and are difficult to treat. This study was undertaken to assess the effect of three antidiabetic plants on the development of cataract in rats. An aqueous extract of Pterocarpus marsupium Linn bark (PM, Hindi name: Vijaysar) (1 g kg(-1) day(-1)), Ocimum sanctum Linn leaves (OS, Hindi name, Tulsi) (200 mg kg(-1) day(-1)) and alcoholic extract of Trigonella foenum-graecum Linn seeds (FG, Hindi name, Methi) (2 g kg(-1) day(-1)) were given to alloxan (120 mg kg(-1)) diabetic rats until the development of cataract. Serum glucose and body weight were monitored at regular intervals while cataract was examined through naked eye as well as slit lamp at 75, 100 and 115 days after alloxan administration. Administration of all the three plant extracts exerted a favorable effect on body weight and blood glucose, the effects were best with PM followed by FG and OS. On the course of cataract development, PM followed by FG exerted anti-cataract effect evident from decreased opacity index while OS failed to produce any anti-cataract effect in spite of significant antihyperglycemic activity.


Lower-Limb-Assisting Robotic Exoskeleton Reduces Energy Consumption in Healthy Young Persons during Stair Climbing.
Many robotic exoskeletons for lower limb assistance aid walking by reducing energy costs. However, investigations examining stair-climbing assistance have remained limited, generally evaluating reduced activation of related muscles. This study sought to investigate how climbing assistance by a robotic exoskeleton affects energy consumption. Ten healthy young participants wearing a robotic exoskeleton that assists flexion and extension of hip and knee joints walked up nine flights of stairs twice at a self-selected speed with and without stair-climbing assistance. Metabolic cost was assessed by measuring oxygen consumption, heart rate, and the time to climb each flight of stairs. Net oxygen cost (NOC) and total heart beats (THB) were used as measures of metabolic cost, accounting for different climbing speeds. Stair-climbing assistance reduced NOC and THB by 9.3% (P < 0.001) and 6.9% (P = 0.003), respectively, without affecting climbing speed. Despite lack of individual optimization, assistive joint torque applied to the hip and knee joints reduced metabolic cost and cardiovascular burden of stair climbing in healthy young males. These results may be used to improve methods for stair ascent assistance.


[Polymorphism of familial cardiovascular changes].
Twenty-five probands (20- to 29-year-old males), selected from an epidemiologic population on the basis of their ECG patterns, and their first-degree relatives were studied. Intrafamilial similarity and difference of cardiovascular patterns were demonstrated, as was their frequent association with hereditary connective-tissue changes.


Spectral and coherence analysis applied to sleep apnea.
The various mechanisms that have been proposed to explain sleep apnea in the elderly are examined. Spectral analysis of the electroencephalogram (EEG), not only in the frequency domain using autoregressive algorithms and the fast Fourier transform, but also in the time domain with comb filters and period-amplitude (PA) analysis, is discussed. The PA features, in particular, may provide an improved basis for the coherence function between ventilation and the state-dependent input.


Domain structure, GTP-hydrolyzing activity and 7S RNA binding of Acidianus ambivalens ffh-homologous protein suggest an SRP-like complex in archaea.
In this study we provide, for the first time, experimental evidence that a protein homologous to bacterial Ffh is part of an SRP-like ribonucleoprotein complex in hyperthermophilic archaea. The gene encoding the Ffh homologue in the hyperthermophilic archaeote Acidianus ambivalens has been cloned and sequenced. Recombinant Ffh protein was expressed in E. coli and subjected to biochemical and functional studies. A. ambivalens Ffh encodes a 50.4-kDa protein that is structured by three distinct regions: the N-terminal hydrophilic N-region (N), the GTP/GDP-binding domain (G) and a C-terminal located C-domain (C). The A. ambivalens Ffh sequence shares 44-46% sequence similarity with Ffh of methanogenic archaea, 34-36% similarity with eukaryal SRP54 and 30-34% similarity with bacterial Ffh. A polyclonal antiserum raised against the first two domains of A. ambivalens Ffh reacts specifically with a single protein (apparent molecular mass: 46 kDa, termed p46) present in cytosolic and in plasmamembrane cell fractions of A. ambivalens. Recombinant Ffh has a melting point of tm = 89 degreesC. Its intrinsic GTPase activity obviously depends on neutral pH and low ionic strength with a preference for chloride and acetate salts. Highest rates of GTP hydrolysis have been achieved at 81 degreesC in presence of 0.1-1 mm Mg2+. GTP hydrolysis is significantly inhibited by high glycerol concentrations, and the GTP hydrolysis rate also markedly decreases by addition of detergents. The Km for GTP is 13.7 microm at 70 degreesC and GTP hydrolysis is strongly inhibited by GDP (Ki = 8 microm). A. ambivalens Ffh, which includes an RNA-binding motif in the C-terminal domain, is shown to bind specifically to 7S RNA of the related crenarchaeote Sulfolobus solfataricus. Comparative sequence analysis reveals the presence of typical signal sequences in plasma membrane as well as extracellular proteins of hyperthermophilic crenarchaea which strongly supposes recognition events by an Ffh containing SRP-like particle in these organisms.


Navigation and Robotics in Spinal Surgery: Where Are We Now?
Spine surgery has experienced much technological innovation over the past several decades. The field has seen advancements in operative techniques, implants and biologics, and equipment such as computer-assisted navigation and surgical robotics. With the arrival of real-time image guidance and navigation capabilities along with the computing ability to process and reconstruct these data into an interactive three-dimensional spinal "map", so too have the applications of surgical robotic technology. While spinal robotics and navigation represent promising potential for improving modern spinal surgery, it remains paramount to demonstrate its superiority as compared to traditional techniques prior to assimilation of its use amongst surgeons.The applications for intraoperative navigation and image-guided robotics have expanded to surgical resection of spinal column and intradural tumors, revision procedures on arthrodesed spines, and deformity cases with distorted anatomy. Additionally, these platforms may mitigate much of the harmful radiation exposure in minimally invasive surgery to which the patient, surgeon, and ancillary operating room staff are subjected.Spine surgery relies upon meticulous fine motor skills to manipulate neural elements and a steady hand while doing so, often exploiting small working corridors utilizing exposures that minimize collateral damage. Additionally, the procedures may be long and arduous, predisposing the surgeon to both mental and physical fatigue. In light of these characteristics, spine surgery may actually be an ideal candidate for the integration of navigation and robotic-assisted procedures.With this paper, we aim to critically evaluate the current literature and explore the options available for intraoperative navigation and robotic-assisted spine surgery.


The role of growth hormone in glucocorticoid-induced osteoporosis.
Glucocorticoid-induced osteoporosis (GIO) is the most common form of secondary osteoporosis. GC influence bone metabolism via a modulation of different components of the GH/IGF-I system. GH has multiple anabolic effects on bone, either direct or mediated by IGF-I. GH-deficient subjects have significant reduction in bone mineralization, bone turnover markers, and increased fracture risk when compared with healthy age-matched controls. The increase of bone remodeling achieved by recombinant human GH (rhGH) therapy may be helpful in both males and females with decreased bone turnover and impaired osteoblastic function such as subjects with GC excess. rhGH treatment may improve lean body mass and skeletal muscle mass that may further reduce fracture risk in GIO patients. Nevertheless, the real efficacy of rhGH and IGF-I treatment in GIO and during aging is still controversial and further well-designed prospective controlled studies are necessary in order to clarify this issue, thus identifying who could potentially benefit from GH treatment.


A phase II study of farnesyl transferase inhibitor R115777 in pancreatic cancer: a Southwest oncology group (SWOG 9924) study.
Ninety per cent of pancreatic adenocarcinomas (PC) contain mutations of the K-Ras proto-oncogene resulting in constitutively activated Ras protein. A critical step in Ras activation is farnesylation of Ras protein. Farnesyl transferase inhibitors are compounds that inhibit farnesylation. We report the results of a phase II trial of R115777, an oral farnesyl transferase inhibitor, in patients with surgically incurable locally advanced or metastatic PC. Between 6/1/2000 and 11/20/2001, 58 cases were accrued, 53 of whom were eligible and analyzable. Patients were required to have a performance status (PS) 0 to 1, be able to take oral medications, and to have adequate renal, hepatic, and hematologic functions. Fifty-five percent were male. Median age was 64.7 years (38.9 to 80.6), and patients had no previous systemic therapy for advanced PC. Treatment consisted of R115777 300 mg po bid given for 3 out of every 4 weeks. Toxicities were as follows: Grade 3 in 19/53 (36%), grade 4 in 53 (173%), and grade 5 in 53 (8%). Most frequent toxicities were: anemia 35/53 (66%), fatigue and malaise 33/53 (62%), nausea 31/53 (58%). Grade 5 toxicities included: thromboembolism 1, infection 2, other 1. Median survival was 2.6 months (mo) (95% CI 2.1-3.6), 6-mo survival is 19% (95% CI, 8-29%), median time to treatment failure was 1.4 mo (95% GI 1.1-1.6). R115777 is ineffective as monotherapy in advanced pancreatic cancer.


Understanding mental toughness in Australian soccer: perceptions of players, parents, and coaches.
We explored mental toughness in soccer using a triangulation of data capture involving players (n = 6), coaches (n = 4), and parents (n = 5). Semi-structured interviews, based on a personal construct psychology (Kelly, 1955/1991) framework, were conducted to elicit participants' perspectives on the key characteristics and their contrasts, situations demanding mental toughness, and the behaviours displayed and cognitions employed by mentally tough soccer players. The results from the research provided further evidence that mental toughness is conceptually distinct from other psychological constructs such as hardiness. The findings also supported Gucciardi, Gordon, and Dimmock's (2009) process model of mental toughness. A winning mentality and desire was identified as a key attribute of mentally tough soccer players in addition to other previously reported qualities such as self-belief, physical toughness, work ethic/motivation, and resilience. Key cognitions reported by mentally tough soccer players enabled them to remain focused and competitive during training and matches and highlighted the adoption of several forms of self-talk in dealing with challenging situations. Minor revisions to Gucciardi and colleagues' definition of mental toughness are proposed.


Tightly linked Rps12 and Rps13 genes provide broad-spectrum Phytophthora resistance in soybean.
The Phytophtora root and stem rot is a serious disease in soybean. It is caused by the oomycete pathogen Phytophthora sojae. Growing Phytophthora resistant cultivars is the major method of controlling this disease. Resistance is race- or gene-specific; a single gene confers immunity against only a subset of the P. sojae isolates. Unfortunately, rapid evolution of new Phytophthora sojae virulent pathotypes limits the effectiveness of an Rps ("resistance to Phytophthora sojae") gene to 8-15 years. The current study was designed to investigate the effectiveness of Rps12 against a set of P. sojae isolates using recombinant inbred lines (RILs) that contain recombination break points in the Rps12 region. Our study revealed a unique Rps gene linked to the Rps12 locus. We named this novel gene as Rps13 that confers resistance against P. sojae isolate V13, which is virulent to recombinants that contains Rps12 but lack Rps13. The genetic distance between the two Rps genes is 4 cM. Our study revealed that two tightly linked functional Rps genes with distinct race-specificity provide broad-spectrum resistance in soybean. We report here the molecular markers for incorporating the broad-spectrum Phytophthora resistance conferred by the two Rps genes in commercial soybean cultivars.


Midwives' and Women's Perception on Moyo Fetal Heart Rate Monitor for Intrapartum Fetal Heart Rate Monitoring; A Cross-Sectional Study.
The annual global neonatal mortality stands at 2.5 million deaths, 1 million of them dying within the first day of life. An additional 2.6 million are stillborn globally, the majority of them due to intrapartum events. Optimal fetal heart rate (FHR) monitoring has the potential to timely detect fetuses at risk and, if coupled with timely obstetric responses may save more newborns. Moyo is a new Doppler with nine crystals capable of monitoring FHR both intermittently and continuously. To assess women's and midwives' opinions on the use of Moyo for intrapartum FHR monitoring. We conducted a cross-sectional study using a structured questionnaire to assess women's and midwives' perception. Women who gave birth at the hospital who used Moyo were interviewed using a questionnaire immediately before discharge from the hospital. Twenty-eight midwives who have been using Moyo for more than 6 months were also interviewed using a structured questionnaire. Data were analyzed using excel and result presented in figures. In total 113 postpartum women who were monitored using Moyo were interviewed before discharge. Out of these, 46 (40.7%) were first-time mothers and the rest were multipara. In total, 95 women (84.1%) used Moyo and other devices for FHR monitoring, 81 (72%) said Moyo was better than Fetoscopes and handheld Doppler, two-third 75 (66.4%) felt that Moyo was comfortable and 93 (82.3%) would like Moyo to be used on them in the future. Out of 28 midwives, 11 (39.3%) used Moyo continuous only, 3 (10.7%) used Moyo intermittently only and 14 (50.0%) used both intermittent and continuous. Thirteen (46.4%) midwives prefer to use Moyo both intermittent and continuous. Sixteen (55.6%) said Moyo was effective, 21 (75%) felt comfortable to use Moyo, and 13 (46.4%) said Moyo was easy to use. The majority of midwives and women who used Moyo felt that Moyo was comfortable for intrapartum FHR monitoring. Moyo can be used both intermittently and continuously depending on the user's preferences.


Noncanonical peptides in complex with MHC class I.
The major histocompatibility complex (MHC) presents canonical medium-high-affinity peptides on the surface of antigen-presenting cells to T-cells. Recognition of peptide MHC by T-cells initiates a cascade of signals which maintains a T-cell-dependent immune response. In the design of vaccines, there is need for an understanding of how peptides bind to MHC class I molecules. Herein, the presentation of canonical anchor motif peptides to MHC class I, noncanonical anchor motif peptides, low-affinity peptides, peptides making use of new pockets, short peptides, long peptides, glycopeptides, retro-inverso peptides and prediction programs for peptides binding to MHC class I molecules is discussed. All this information will aid in the design of new and improved peptide-based vaccines.


Older people's views of quality of care: a randomised controlled study of continuum of care.
To analyse frail older people's views of quality of care when receiving a comprehensive continuum of care intervention, compared with those of people receiving the usual care (control group). The intervention included early geriatric assessment, case management, interprofessional collaboration, support for relatives and organising of care-planning meetings in older people's own homes. Prior studies indicate that tailored/individualised care planning conducted by a case manager/coordinator often led to greater satisfaction with care planning among older people. However, there is no obvious evidence of any effects of continuum of care interventions on older people's views of quality of care. Randomised controlled study. Items based on a validated questionnaire were used in face-to-face interviews to assess older people's views of quality of care at three, six and 12 months after baseline. Older people receiving a comprehensive continuum of care intervention perceived higher quality of care on items about care planning (p ≤ 0·005), compared with those receiving the usual care. In addition, they had increased knowledge of whom to contact about care/service, after three and 12 months (p < 0·03). The study gives evidence of the advantages of a combination of components such as organising care-planning meetings in older people's own homes, case management and interprofessional teamwork. The results have implications for policymakers, managers and professionals in the area of health and social care for older people to meet individual needs of frail older people.


Effects of incentivizing viral suppression in previously incarcerated adults living with HIV.
Background: The amount of HIV in a person's blood can be suppressed to an undetectable level through antiretroviral therapy medications (ART). Adhering to an ART regimen can improve a person's health and reduce HIV transmission. Despite these benefits, many people with HIV do not maintain the level of adherence required to achieve an undetectable viral load. This problem is particularly common among people who have been incarcerated.Objective: To determine effects of incentivizing viral suppression in previously incarcerated adults with HIV.Methods: Adults with HIV (N = 102) and detectable viral load (>200 copies/mL) were randomly assigned to a Usual Care or Incentive group. Usual Care participants did not earn incentives for viral suppression. Incentive participants earned incentives ($10/day maximum) for providing blood samples with a reduced or undetectable (<200 copies/mL) viral load. Assessments were conducted every 3 months. Results collected during the first year were aggregated and compared based on group assignment and incarceration history.Results: Previously incarcerated participants in the Incentive group provided more (OR: 2.9; CI: 1.3-6.8; p <.05) blood samples with an undetectable viral load (69%) than those in the Usual Care group (41%). Never-incarcerated participants in the Incentive group provided more (OR: 6.8; CI: 2.2-21.0; p <.01) blood samples with an undetectable viral load (78%) than those in the Usual Care group (36%). Effects of incentives did not differ by incarceration history.Conclusions: Incentivizing viral suppression can increase viral suppression (undetectable viral load) in people who have been incarcerated.


Purification and characterization of insulin from the Australian lungfish, Neoceratodus forsteri (Dipnoi).
The Australian lungfish Neoceratodus forsteri, a facultative air breather, is considered to be the most primitive of the extant Dipnoi and so occupies a uniquely important evolutionary position in the transition from fish to tetrapods. Insulin was isolated from an extract of the pancreas of N. forsteri and its primary structure established as: A-Chain, Gly-Ile-Val-Glu-Gln-Cys-Cys-His-Thr-Pro(10)-Cys-Ser-Leu-Tyr-Gln-Leu-G lu-Asn-Tyr-Cys(20)-Asn-Glu-Thr-Glu; B-Chain, Ala-Ala-Val-Asn-Gln-His-Leu-Cys-Gly-Ser(10)-His-Leu-Val-Glu-Ala-Leu- Tyr-Phe-Val-Cys(20)-Gly-Glu-Arg-Gly-Phe-Phe-Tyr-Leu-Pro- Lys(30)-Gly. This amino acid sequence is more similar to that of human insulin than to insulins from present-day amphibians. All the residues in human insulin that are considered to be important in receptor binding, dimerization, and hexamerization are conserved in lungfish insulin except for the substitution (Leu --> Phe) at the position corresponding to B17 in human insulin. Consistent with the assertion that the Dipnoi is a monophyletic group, insulins from N. forsteri and from the African lungfish Protopterus annectens contain extensions to the C-terminus of the A-chain and to the N-terminus of the B-chain that have not been found in other sarcopterygian species. However, the unusual amino acid substitutions found in insulin from P. annectens (e.g., GlyB21 --> Ala, GluB22 --> Asp, and ArgB23 --> Asn) are not present in N. forsteri insulin, suggesting that they occurred in the Protopterus lineage after divergence of the genera.


Myeloid madness: assessing diagnostic inconsistency between the new WHO and ICC schemes for myelodysplastic/myeloproliferative neoplasms.
The classification of haematological neoplasms recently underwent revision, generating two separate schemes-the International Consensus Classification and the fifth edition of the WHO classification. The new division into separate classification systems presents challenges for haematopathologists, haematologists/oncologists and patients. While it is too early to assess the full clinical impact, we sought to identify diagnostic discordance which may arise from applying separate classification schemes in myeloid neoplasia, and particularly in the challenging category of myelodysplastic syndrome/myeloproliferative neoplasms. A review of 64 such cases found 1 case with a significant discrepancy between the WHO and International Consensus Classification systems, and 9 cases with nominal discrepancies. Confusion from the use of conflicting diagnostic terms represents a potential source of patient harm, increased pathologist workload and burnout and erosion of clinician and patient trust.


[Alpha-1-antitrypsin and cystic fibrosis (author's transl)].
A methodical study of the alpha-1-antitrypsin serum level and the Pi type in 69 CF children, 156 CF heterozygote parents and 200 blood donors from Paris, has been made. Although the PiZ allele frequency was found to be higher in the CF population, especially in fathers, than in the reference population, the difference is not statistically relevant. Therefore there does not seem to be a relation between cystic fibrosis and the Pi alleles system.


Elucidating the individual effects of calcium and phosphate ions on hMSCs by using composite materials.
The biological performance of bone graft substitutes based on calcium phosphate bioceramics is dependent on a number of properties including chemical composition, porosity and surface micro- and nanoscale structure. However, in contemporary bioceramics these properties are interlinked, therefore making it difficult to investigate the individual effects of each property on cell behavior. In this study we have attempted to investigate the effects of calcium and inorganic phosphate ions independent from one another by preparing composite materials with polylactic acid (PLA) as a polymeric matrix and calcium carbonate or sodium phosphate salts as fillers. Clinically relevant bone marrow derived human mesenchymal stromal cells (hMSCs) were cultured on these composites and proliferation, osteogenic differentiation and ECM mineralization were investigated with time and were compared to plain PLA control particles. In parallel, cells were also cultured on conventional cell culture plates in media supplemented with calcium or inorganic phosphate to study the effect of these ions independent of the 3D environment created by the particles. Calcium was shown to increase proliferation of cells, whereas both calcium and phosphate positively affected alkaline phosphatase enzyme production. QPCR analysis revealed positive effects of calcium and of inorganic phosphate on the expression of osteogenic markers, in particular bone morphogenetic protein-2 and osteopontin. Higher levels of mineralization were also observed upon exposure to either ion. Effects were similar for cells cultured on composite materials and those cultured in supplemented media, although ion concentrations in the composite cultures were lower. The approach presented here may be a valuable tool for studying the individual effects of a variety of soluble compounds, including bioinorganics, without interference from other material properties.


Common functional networks in the mouse brain revealed by multi-centre resting-state fMRI analysis.
Preclinical applications of resting-state functional magnetic resonance imaging (rsfMRI) offer the possibility to non-invasively probe whole-brain network dynamics and to investigate the determinants of altered network signatures observed in human studies. Mouse rsfMRI has been increasingly adopted by numerous laboratories worldwide. Here we describe a multi-centre comparison of 17 mouse rsfMRI datasets via a common image processing and analysis pipeline. Despite prominent cross-laboratory differences in equipment and imaging procedures, we report the reproducible identification of several large-scale resting-state networks (RSN), including a mouse default-mode network, in the majority of datasets. A combination of factors was associated with enhanced reproducibility in functional connectivity parameter estimation, including animal handling procedures and equipment performance. RSN spatial specificity was enhanced in datasets acquired at higher field strength, with cryoprobes, in ventilated animals, and under medetomidine-isoflurane combination sedation. Our work describes a set of representative RSNs in the mouse brain and highlights key experimental parameters that can critically guide the design and analysis of future rodent rsfMRI investigations.


Subsensitivity of serotonin receptors after long-term treatment of rats with fluoxetine.
Binding of [3H]-serotonin ([3H]5-hydroxytryptamine, [3H]5HT) to high and low affinity sites in brain membranes from frontal cortex was compared in rats fed control chow or fluoxetine (0.01%) containing chow. A reduction in the number of high affinity binding sites and in the affinity of the low affinity binding sites was observed after 2, 4 or 6 weeks. Reduction in the number of [3H]5HT binding sites was also observed in cerebral cortex of rats given fluoxetine by daily intraperitoneal injection for 4 or 6 weeks. Thus, long-term treatment with a selective inhibitor of serotonin uptake, fluoxetine, caused subsensitivity of 5HT receptors in frontal cortex and cerebral cortex of rat brain.


Pavlovian conditioning of emotional responses to olfactory and contextual stimuli: a potential model for the development and expression of chemical intolerance.
Chemical intolerance (CI) in humans is a poorly understood phenomenon of uncertain etiology, seemingly influenced by multiple factors both within and between affected individuals. Several authors have suggested that the development of CI in some individuals may be due, at least in part, to Pavlovian conditioning processes in which the expression of overt symptoms to certain substances reflects classically conditioned responses to previously neutral olfactory and contextual stimuli. In this paper, we describe the potential relationship between olfactory and contextual conditioning in experimental animals and the development and expression of CI in humans. Furthermore, as significant advances have been made in delineating the brain areas that underlie these learned responses, we also review recent research on the contributions of the amygdala and perirhinal cortical region to olfactory and contextual fear conditioning.


Relationship Between Personality Disorders Scales, Pathological Personality Traits, and Six Domains of Functioning in Sample With Alcohol Use Disorder.
Studies reveal a functional impairment in patients with personality disorders (PDs), but there is not enough information to form conclusions about this relation in patients with alcohol use disorder (AUD). The aim of this study was to investigate to what extent a personality disorders scales including pathological personality traits (PPTs) predict six domains of functioning in patients with AUD. In total, 48 patients with AUD diagnosis, who were treated in the psychiatric clinics, aged 20 to 65 years [M = 37.5; SD = 12.08; 12 (25%) females and 36 (75%) males], filled out the demographic questionnaire, WHO Disability Assessment Schedule 2.0 (WHODAS 2.0, Latvian version) and Latvian Clinical Personality Inventory (LCPI v2.1.). All respondents signed the informed consent form. Stepwise regression analysis showed that PD Avoidant scale positively predicts impairment in Cognition and Getting along domains of functioning in AUD patients, but, on the PPTs level, it was found that Social withdrawal along with Irresponsibility and Guilt/Shame positively predict impairment in Cognition domain of functioning, and Social withdrawal along with Depressivity and Irresponsibility positively predict impairment in Getting along domain of functioning. The results of the study showed that PPT Orderliness negatively predicts impairment in Live activities domain of functioning. The PD Dependent scale and PPT Separation insecurity positively predict impairment in Participation domain of functioning. Obtained results add deeper insight into understanding of the relationship between personality disorders scales including pathological personality traits and six domains of functioning in patients with AUD.


What is craving? Models and implications for treatment.
Although many alcoholics experience craving, researchers have not yet developed a common, valid definition of the phenomenon. Numerous models of the mechanisms underlying craving have been suggested, however. One of those models--the neuroadaptive model--suggests that the prolonged presence of alcohol induces changes in brain-cell function. In the absence of alcohol, those changes cause an imbalance in brain activity that results in craving. Furthermore, the adaptive changes generate memories of alcohol's pleasant effects that can be activated when alcohol-related environmental stimuli are encountered, even after prolonged abstinence, thereby leading to relapse. Similarly, stressful situations may trigger memories of the relief afforded by alcohol, which could also lead to relapse. Neurobiological and brain-imaging studies have identified numerous brain chemicals and brain regions that may be involved in craving. Psychiatric conditions that affect some of these brain regions, such as depression or anxiety, also may influence craving. A better understanding and more reliable assessment of craving may help clinicians tailor treatment to the specific needs of each patient, thereby reducing the risk of relapse.


Identification of diverse Salmonella serotypes, virulotypes, and antimicrobial resistance phenotypes in waterfowl from Chile.
Salmonella enterica is a pathogen with a wide host-range that presents great concern in developed and developing countries. To determine and characterize Salmonella strains found in Chile's waterfowl, we sampled 758 birds along 2000 km of the Chilean coast. In this sample, 46 isolates from 10 serotypes were detected, several with multidrug resistance phenotypes and different combinations of virulence-associated genes (virulotypes). These results suggest that Salmonella infection in waterfowl in Chile could have impacts on public and animal health.


Microgel-based inks for paper-supported biosensing applications.
As a first step for the development of biosensing inks for inexpensive paper-based biodetection, we prepared paper strips printed with carboxylic poly( N-isopropylacrylamide) microgels that were modified either with an antibody or with a DNA aptamer. We found that the antibody and the DNA aptamer retained their recognition capabilities when coupled to microgel. The printed microgel remains stationary during chromatographic elution while the microgel-supported molecular recognition elements are accessible to their intended targets present in the elution solution. Our work indicates that microgels, large enough to isolate the biosensors from the paper surface, are sufficiently hydrophilic to be wetted during chromatographic elution, exposing the gel-supported affinity probes to their targets.


Selective stimulation of the spinal cord surface using a stretchable microelectrode array.
By electrically stimulating the spinal cord, it is possible to activate functional populations of neurons that modulate motor and sensory function. One method for accessing these neurons is via their associated axons, which project as functionally segregated longitudinal columns of white-matter funiculi (i.e., spinal tracts). To stimulate spinal tracts without penetrating the cord, we have recently developed technology that enables close-proximity, multi-electrode contact with the spinal cord surface. Our stretchable microelectrode arrays (sMEAs) are fabricated using an elastomer polydimethylsiloxane substrate and can be wrapped circumferentially around the spinal cord to optimize electrode contact. Here, sMEAs were used to stimulate the surfaces of rat spinal cords maintained in vitro, and their ability to selectively activate axonal surface tracts was compared to rigid bipolar tungsten microelectrodes pressed firmly onto the cord surface. Along dorsal column tracts, the axonal response to sMEA stimulation was compared to that evoked by rigid microelectrodes through measurement of their evoked axonal compound action potentials (CAPs). Paired t-tests failed to reveal significant differences between the sMEA's and the rigid microelectrode's stimulus resolution, or in their ranges of evoked CAP conduction velocities. Additionally, dual-site stimulation using sMEA electrodes recruited spatially distinct populations of spinal axons. Site-specific stimulation of the ventrolateral funiculus - a tract capable of evoking locomotor-like activity - recruited ventral root efferent activity that spanned several spinal segments. These findings indicate that the sMEA stimulates the spinal cord surface with selectivity similar to that of rigid microelectrodes, while possessing potential advantages concerning circumferential contact and mechanical compatibility with the cord surface.


EQ-5D-5L Valuation for the Malaysian Population.
The aim of this study was to develop an EQ-5D-5L value set reflecting the health preferences of the Malaysian adult population. Respondents were sampled with quotas for urbanicity, gender, age, and ethnicity to ensure representativeness of the Malaysian population. The study was conducted using a standardized protocol involving the EuroQol Valuation Technology (EQ-VT) computer-assisted interview system. Respondents were administered ten composite time trade-off (C-TTO) tasks and seven discrete choice experiment (DCE) tasks. Both linear main effects and constrained non-linear regression models of C-TTO-only data and hybrid models combining C-TTO and DCE data were explored to determine an efficient and informative model for value set prediction. Data from 1125 respondents representative of the Malaysian population were included in the analysis. Logical consistency was present in all models tested. Using cross-validation, eight-parameter models for C-TTO only and C-TTO + DCE hybrid data displayed greater out-of-sample predictive accuracy than their 20-parameter, main-effect counterparts. The hybrid eight-parameter model was chosen to represent the Malaysian value set, as it displayed greater out-of-sample predictive accuracy over C-TTO data than the C-TTO-only model, and produced more precise estimates. The estimated value set ranged from - 0.442 to 1. The constrained eight-parameter hybrid model demonstrated the best potential in representing the Malaysian value set. The presence of the Malaysian EQ-5D-5L value set will facilitate its application in research and health technology assessment activities.


Effects of diabetes mellitus and glycemic traits on cardiovascular morpho-functional phenotypes.
The effects of diabetes on the cardiac and aortic structure and function remain unclear. Detecting and intervening these variations early is crucial for the prevention and management of complications. Cardiovascular magnetic resonance imaging-derived traits are established endophenotypes and serve as precise, early-detection, noninvasive clinical risk biomarkers. We conducted a Mendelian randomization (MR) study to examine the association between two types of diabetes, four glycemic traits, and preclinical endophenotypes of cardiac and aortic structure and function. Independent genetic variants significantly associated with type 1 diabetes, type 2 diabetes, fasting insulin (FIns), fasting glucose (FGlu), 2 h-glucose post-challenge (2hGlu), and glycated hemoglobin (HbA1c) were selected as instrumental variables. The 96 cardiovascular magnetic resonance imaging traits came from six independent genome-wide association studies. These traits serve as preclinical endophenotypes and offer an early indication of the structure and function of the four cardiac chambers and two aortic sections. The primary analysis was performed using MR with the inverse-variance weighted method. Confirmation was achieved through Steiger filtering and testing to determine the causal direction. Sensitivity analyses were conducted using the weighted median, MR-Egger, and MR-PRESSO methods. Additionally, multivariable MR was used to adjust for potential effects associated with body mass index. Genetic susceptibility to type 1 diabetes was associated with increased ascending aortic distensibility. Conversely, type 2 diabetes showed a correlation with a reduced diameter and areas of the ascending aorta, as well as decreased distensibility of the descending aorta. Genetically predicted higher levels of FGlu and HbA1c were correlated with a decrease in diameter and areas of the ascending aorta. Furthermore, higher 2hGlu levels predominantly showed association with a reduced diameter of both the ascending and descending aorta. Higher FIns levels corresponded to increased regional myocardial-wall thicknesses at end-diastole, global myocardial-wall thickness at end-diastole, and regional peak circumferential strain of the left ventricle. This study provides evidence that diabetes and glycemic traits have a causal relationship with cardiac and aortic structural and functional remodeling, highlighting the importance of intensive glucose-lowering for primary prevention of cardiovascular diseases.


Uveitis-glaucoma-hyphema syndrome after handmade, anterior chamber lens implantation.
Two patients developed uveitis-glaucoma-hyphema syndrome secondary to implantation of handmade anterior chamber intraocular lenses (IOLs). The IOLs were designed and made by the patients' ophthalmologists in their native Ukraine. The cases illustrate innovative and unusual solutions to the shortage of quality IOLs in the developing world and the challenging management of patients who present with complications arising from these IOLs.


Shared decision-making at end-of-life is aided by graphical trending of illness severity.
The Rothman Index (RI) gives a visual picture of patient's condition and progress for the physician and family to view together. This case demonstrates how the RI graph facilitates physician-family communication. An 85-year-old man with normal pressure hydrocephalus and ventriculoperitoneal shunt presented with a subdural haematoma. He required a temporoparietal craniotomy and evacuation of left subdural haematoma, followed by care in an intensive inpatient rehabilitation unit. His course was complicated by aspiration pneumonia, dehydration, renal failure and phenytoin toxicity. During hospitalisation, the patient's RI graph was reviewed daily with his family. The RI provided an unambiguous visualisation of the trend of patient acuity, which depicted the patient's persistent decline in health, and made clear to the family the situation of the patient. This clarity was instrumental in prompting frank discussions of prognosis and consideration of comfort measures, resulting in timely transfer to hospice.


Antioxidant evaluation and oxidative stability of structured lipids from extravirgin olive oil and conjugated linoleic acid.
Structured lipid (SL) was synthesized from extravirgin olive oil (EVOO) and conjugated linoleic acid (CLA) via a lipase-catalyzed reaction. CLA provides a variety of health benefits, but it is not consumed in free fatty acid form. The synthesized SL olive oil contained 42.5 mol % CLA isomers, and the major isomers were cis-9,trans-11-CLA (16.9 mol %) and trans-10,cis-12-CLA (24.2 mol %). The antioxidant activity determined by the radical scavenging capacity with the 2,2-diphenyl-1-picrylhydrazyl radical was lower in SL olive oil than in EVOO. The oxidative stability was also lower in SL olive oil since it had a higher peroxide value, rho-anisidine value, and 2-thiobarbituric acid reactive substances values during 20 days of storage at 60 degrees C. This observation could be due to the reduction in the natural phenolic compounds (97%) and tocopherols (56%), and the incorporated CLA with two conjugated double bonds in the SL olive oil. The oxidative stability of SL olive oil was increased by added rosemary extracts at concentrations of 100, 200, and 300 ppm. The present study suggests that the SL olive oil may be a suitable way to incorporate or deliver CLA into human diets. However, the addition of a proper antioxidant would be required for improving its oxidative stability.


Protective effect of miR-18a in resected liver metastases of colorectal cancer and FOLFOX treatment.
Colorectal cancer ranks second in terms of cancer associated deaths worldwide, whereas miRNA play a pivotal role in the etiology of cancer and its metastases. Studying the expression and cellular function of miR-18a in metastatic colorectal cancer and association to progression-free survival. Colorectal liver metastases (N = 123) and primary colorectal cancer (N = 27) where analyzed by RT-PCR and correlated with clinical follow up data. Invasion and migration assays were performed with the liver metastatic cell line LIM2099 after miR-18a knockdown. Cell viability under FOLFOX treatment and knockdown was measured. We found that the expression of miR-18a was increased 4.38-fold in liver metastases and 3.86-fold in colorectal tumor tissue compared to healthy liver tissue and colorectal mucosa, respectively (p ≤ .001). Patients with a high miR-18a expression in liver metastases had a progression-free survival (PFS) of 13.6 months versus 8.9 months in patients with low expression (N = 123; p = .024). In vitro migration of LIM2099 cells was reduced after miR-18a knockdown and cell viability was significantly increased after miR-18a knockdown and treatment with folinic acid or oxaliplatin. Subgroup analysis of PFS revealed significant benefits for patients with high miR-18a expression receiving 5-FU, folinic acid or oxaliplatin. High expression of miR-18a in colorectal liver metastases might have a protective effect after resection of metastases and FOLFOX treatment regarding PFS.


Adverse Effects Under Th2 Biologics (Dupilumab and Tezepelumab) in a Patient with Severe Asthma and Atopic Dermatitis.
This is the first report of a patient with severe asthma and atopic dermatitis that developed local perioral skin infection which onset coincided with the patient's treatment with dupilumab (candida albicans) and later with tezepelumab (microscopic detection of yeast). Besides moderate headache, macular exanthema was found after administration of tezepelumab, which was subsequently accompanied by a worsening of symptoms upon reexposure to the treatment. Both sensations needed multidisciplinary treatment and both antibody therapies were stopped.


Acceptability of Psychosis Screening and Factors Affecting Its Implementation: Interviews With Community Health Care Providers.
Although screening for psychosis may reduce the duration of untreated psychosis, the barriers and facilitators associated with implementing such a procedure in various care settings have not been explored. Investigators conducted in-depth, semistructured interviews with 17 members of school counseling services or community mental health staff at sites that administer a psychosis screening tool. Using an inductive approach to thematic analysis, they evaluated the acceptability of psychosis screening and barriers to and facilitators of implementation. Participants reported few barriers to implementation. However, several service-, client-, and program-level factors were considered to significantly affect the implementation of screening. Most participants found that using the screening tool did not significantly affect their overall workload. Facilitators included leadership support, the novelty of using a technology-based screener, regular staff training, and the importance of establishing an effective link between community services and specialty care, with these factors important at different stages of the process. Screening for psychosis was associated with significant advantages over referrals based on clinical judgment alone, including increased speed and accuracy of identification, increased confidence in diagnosis, and the provision of a clear pathway to specialty treatment. The experiences of school counseling and community mental health teams suggest that incorporating a technology-based screening procedure for early psychosis is feasible. Identifying barriers and facilitators at various stages of the screening procedure may reduce the dropout of clients potentially eligible for early psychosis care.


Expression of p19INK4d, CDK4, CDK6 in glioblastoma multiforme.
Deregulation of the G1/S checkpoint is a frequent event in the development of glioblastoma multiforme (GBM). Previous studies have shown more than 50% of primary GBM tumours contain either complete loss of the p16INK4a locus or amplification of the CDK4 gene. Moreover, many heterozygosity studies have shown deletion on human chromosome 19p13.2, where the p19INK4d gene has been localized. We examined the expression of p19INK4d and its two CDK substrates in a series of glioma-derived cell lines and tumours. No gene rearrangement or deletion was observed in the p19INK4d gene in these cell lines; however, expression of CDK4 and CDK6 was elevated relative to matched normal brain tissue in eight of 18 GBM tumours (44%). Furthermore, CDK6 expression level was increased in 12/14 glioblastomas, but undetectable in tumour samples of a previous lower grade tumour from the same patient. These data attest to the functional importance of both CDK4 and CDK6 in astrocytic tumourigenesis, particularly during the later stages of tumour progression.


Synthesis of fused piperidinones through a radical-ionic cascade.
Azabicyclo[4.3.0]nonanes were assembled, from chiral allylsilanes possessing an oxime moiety, using a stereocontrolled formal [2 + 2 + 2] radical-ionic process. The cascade involves the addition of an alpha-iodoester to the less substituted end of the enoxime which is then followed by a 5-exo-trig cyclization onto the aldoxime function, producing an alkoxyaminyl radical species which finally lactamizes to afford the titled piperidinone. High levels of stereoinduction were observed, demonstrating the ability of a silicon group located at the allylic position to efficiently control the stereochemistry of the two newly created stereogenic centers. When the radical cascade was extended to ketoximes, the resulting sterically hindered alkoxyaminyl radical did not react further with the initiator Et3B to produce the expected nucleophilic amidoborane complex. In sharp contrast, this long-lived radical recombined with the initial alpha-stabilized ester radical to produce a cyclopentane incorporating two ester fragments.


Clinical characteristics and predictors of burn complicated with smoke inhalation injury: A retrospective analysis.
Fire smoke enters the human lungs through the respiratory tract. The damage to the respiratory tract and lung tissue is known as smoke inhalation injury (SII). Fire smoke can irritate airway epithelium cells, weaken endothelial cell adhesion and lyse alveolar type II epithelia cells, leading to emphysema, decreased lung function, pneumonia and risk of acute lung injury/acute respiratory distress syndrome (ARDS). The purpose of the present study was to analyze the clinical characteristics of patients with SII and the risk factors affecting their prognosis. A total of 103 patients with SII admitted between January 2016 to December 2021 to the Burns Unit of the Characteristic Medical Center of Chinese People's Armed Police Force and 983 Hospital of the Joint Logistics Support Force of the Chinese People's Liberation Army were selected for the present study. The demographics and clinical features between different severities of SII were analyzed. Univariate/multivariate logistic regression was used to analyze the potential predictors for severity, ARDS and mortality of patients with SII. Receiver operating characteristic (ROC) curves were used to screen independent risk factors and identify their prediction accuracy. It was concluded that total body surface area (TBSA), III burn area (of total %TBSA), cases of respiratory infections, ARDS morbidity, mortality, acute physiology and chronic health evaluation II, lung injury prediction score, lactic acid, white blood cells (WBC), alanine transaminase, blood urea nitrogen, serum creatinine and uric acid were indicators that were raised with increasing severity of SII. However red blood cells, hemoglobin, platelet count, total protein, albumin, and albumin/globulin were decreased with the increasing severity of SII (P<0.05). WBC >20.91 (109/l) was a reliable indicator for severe SII. Lactic acid >9.60 (mmol/l) demonstrated a high degree of accuracy in predicting ARDS development in patients with SII. Hemoglobin <83.00 (g/l) showed a high degree of accuracy in predicting mortality. In summary, the highlighted assessment parameters could be used to contribute to devising improved treatment plans to preempt worsening conditions (such as shock, ARDS, multiple organ dysfunction syndrome and death).


Glyphosate inhibits melanization and increases susceptibility to infection in insects.
Melanin, a black-brown pigment found throughout all kingdoms of life, has diverse biological functions including UV protection, thermoregulation, oxidant scavenging, arthropod immunity, and microbial virulence. Given melanin's broad roles in the biosphere, particularly in insect immune defenses, it is important to understand how exposure to ubiquitous environmental contaminants affects melanization. Glyphosate-the most widely used herbicide globally-inhibits melanin production, which could have wide-ranging implications in the health of many organisms, including insects. Here, we demonstrate that glyphosate has deleterious effects on insect health in 2 evolutionary distant species, Galleria mellonella (Lepidoptera: Pyralidae) and Anopheles gambiae (Diptera: Culicidae), suggesting a broad effect in insects. Glyphosate reduced survival of G. mellonella caterpillars following infection with the fungus Cryptococcus neoformans and decreased the size of melanized nodules formed in hemolymph, which normally help eliminate infection. Glyphosate also increased the burden of the malaria-causing parasite Plasmodium falciparum in A. gambiae mosquitoes, altered uninfected mosquito survival, and perturbed the microbial composition of adult mosquito midguts. Our results show that glyphosate's mechanism of melanin inhibition involves antioxidant synergy and disruption of the reaction oxidation-reduction balance. Overall, these findings suggest that glyphosate's environmental accumulation could render insects more susceptible to microbial pathogens due to melanin inhibition, immune impairment, and perturbations in microbiota composition, potentially contributing to declines in insect populations.


Analysis of 1,000 Type-Strain Genomes Improves Taxonomic Classification of Bacteroidetes.
Although considerable progress has been made in recent years regarding the classification of bacteria assigned to the phylum Bacteroidetes, there remains a need to further clarify taxonomic relationships within a diverse assemblage that includes organisms of clinical, piscicultural, and ecological importance. Bacteroidetes classification has proved to be difficult, not least when taxonomic decisions rested heavily on interpretation of poorly resolved 16S rRNA gene trees and a limited number of phenotypic features. Here, draft genome sequences of a greatly enlarged collection of genomes of more than 1,000 Bacteroidetes and outgroup type strains were used to infer phylogenetic trees from genome-scale data using the principles drawn from phylogenetic systematics. The majority of taxa were found to be monophyletic but several orders, families and genera, including taxa proposed long ago such as Bacteroides, Cytophaga, and Flavobacterium but also quite recent taxa, as well as a few species were shown to be in need of revision. According proposals are made for the recognition of new orders, families and genera, as well as the transfer of a variety of species to other genera. In addition, emended descriptions are given for many species mainly involving information on DNA G+C content and (approximate) genome size, both of which can be considered valuable taxonomic markers. We detected many incongruities when comparing the results of the present study with existing classifications, which appear to be caused by insufficiently resolved 16S rRNA gene trees or incomplete taxon sampling. The few significant incongruities found between 16S rRNA gene and whole genome trees underline the pitfalls inherent in phylogenies based upon single gene sequences and the impediment in using ordinary bootstrapping in phylogenomic studies, particularly when combined with too narrow gene selections. While a significant degree of phylogenetic conservation was detected in all phenotypic characters investigated, the overall fit to the tree varied considerably, which is one of the probable causes of misclassifications in the past, much like the use of plesiomorphic character states as diagnostic features.


State-based SHOSLIF for indoor visual navigation.
In this paper, we investigate vision-based navigation using the self-organizing hierarchical optimal subspace learning and inference framework (SHOSLIF) that incorporates states and a visual attention mechanism. With states to keep the history information and regarding the incoming video input as an observation vector, the vision-based navigation is formulated as an observation-driven Markov model (ODMM). The ODMM can be realized through recursive partitioning regression. A stochastic recursive partition tree (SRPT), which maps an preprocessed current input raw image and the previous state into the current state and the next control signal, is used for efficient recursive partitioning regression. The SRPT learns incrementally: each learning sample is learned or rejected "on-the-fly." The purposed scheme has been successfully applied to indoor navigation.


Achievements and objectives for 2019.
At the beginning of 2019 we wish to inform our readers and share with them the achievements and objectives of the Editorial Board. With the gradual growth of the journal, and to be more efficient and better serve the scientific societies it represents (Spanish Society of Gastroenterology [SEPD], Spanish Society of Gastrointestinal Endoscopy [SEED] and Spanish Association of Digestive Ultrasonography [AEED]), we made the decision to progressively increase the number of papers per issue, and to adapt manuscript classification to the requirements of Web of Science. The number of original papers was increased to six, and we look forward to having a total of eight, with two clinical notes and two images in digestive disorders, and keeping one review and/or special article per issue.


Cholera toxin: an intracellular journey into the cytosol by way of the endoplasmic reticulum.
Cholera toxin (CT), an AB(5)-subunit toxin, enters host cells by binding the ganglioside GM1 at the plasma membrane (PM) and travels retrograde through the trans-Golgi Network into the endoplasmic reticulum (ER). In the ER, a portion of CT, the enzymatic A1-chain, is unfolded by protein disulfide isomerase and retro-translocated to the cytosol by hijacking components of the ER associated degradation pathway for misfolded proteins. After crossing the ER membrane, the A1-chain refolds in the cytosol and escapes rapid degradation by the proteasome to induce disease by ADP-ribosylating the large G-protein Gs and activating adenylyl cyclase. Here, we review the mechanisms of toxin trafficking by GM1 and retro-translocation of the A1-chain to the cytosol.


Pheochromocytoma: An overlooked reversible cause of heart failure with reduced ejection fraction.
Pheochromocytoma is a rare, benign tumor of adrenal medulla, and its clinical symptoms are related to catecholamine production. Clinical presentation may vary in a broad spectrum. Dilated or hypertrophied cardiomyopathies are the possible clinical outcomes of pheochromocytoma. Pheochromocytoma should be kept in mind in the differential diagnosis where resistant hypertension, palpitations, headache, and sweating accompany cardiomyopathy. Excessive adrenergic stimulation causing catecholamine discharge can trigger hypertension crisis, pulmonary edema, and myocardial necrosis. Here in this report, we aimed to present the case of a patient with pheochromocytoma related cardiomyopathy who was totally recovered after surgical excision of the tumor.


Tentative breakpoints and areas of technical uncertainty for early reading automated disc diffusion for Enterobacterales.
Disc diffusion is a reliable, accurate and cost-efficient procedure for antimicrobial susceptibility testing (AST) but requires long (18-24 h) incubation times. Reading of disc diffusion after short incubation times (6-8 h) by automated systems is feasible but should be categorized with time-adapted breakpoints to reduce errors. This study systematically compared early readings (6 and 8 h) of disc diffusion using an automated system with that of the standard 18 h EUCAST method. Time-adapted tentative breakpoints were proposed to discriminate susceptible from resistant isolates and areas of technical uncertainty were defined to minimize the risk of errors. A total of 1106 Enterobacterales isolates with a wide variety of resistance mechanisms and resistance profiles were included. All isolates were analysed for susceptibility to amoxicillin/clavulanic acid, ceftriaxone, cefepime, meropenem, ciprofloxacin and gentamicin using the automated WASPLabTM system. Part of the collection (515 isolates) was also analysed for susceptibility to an additional 10 antibiotics. Separation between WT and non-WT populations was poorer at early incubation times than following standard incubation. Editing of rapid automated AST results after 6 and 8 h incubation with time-adapted breakpoints resulted in 84.0% and 88.5% interpretable results with assignment to the resistant or susceptible category. Major error and very major error rates for the 6 h readings were only 0.4% and 0.3%, virtually identical to those of 18 h AST reading. Time-adapted clinical breakpoints in disc diffusion testing for Enterobacterales allow for accurate automated AST interpretation after shortened incubation times for a large number of antibiotics, with the additional possibility of subsequent confirmation after 18 h incubation.


Pentoxifylline Decreases Dialysis Risk in Patients With Advanced Chronic Kidney Disease.
Few studies evaluated the effects of pentoxifylline on hard endpoints in patients with predialysis stage 5 chronic kidney disease (CKD). Thus, we tried to explore the effects of pentoxifylline and its interaction with renin-angiotensin-aldosterone system (RAAS) blockade on the development of endstage renal disease (ESRD) and mortality. This nationwide cohort study retrospectively included patients who had a serum creatinine level of >6 mg/dL and received erythropoiesis-stimulating agents (ESAs) between 2000 and 2010. We analyzed 7,366 pentoxifylline users and 7,366 propensity score-matched nonusers. Using Cox proportional hazard models, pentoxifylline reduced the risks of ESRD and the composite renal outcome but not that of mortality. In terms of the risks of developing ESRD, pentoxifylline alone exerted a comparable beneficial effect to combined therapy with an RAAS inhibitor and greater renoprotection than RAAS inhibitor monotherapy. This study suggests pentoxifylline is efficacious in slowing progression to ESRD in patients with predialysis stage 5 CKD.


[Experience of microbiologic diagnostics of acute pneumonia cases arose in town Verkhnyaya Pyshma in July-August 2007].
Bacteriologic, serologic, and PCR tests of blood, sputum, serum and autopsy samples from 91 patients were performed during outbreak of pneumonia in town Verkhnyaya Pyshma in July-August 2007. Streptococcus pneumoniae was isolated from 20% of diagnostically meaningful samples of sputum and from 50% of autopsy samples. Diagnosis of Legionnaires' disease was confirmed in 48% of cases. It was shown that association of Streptococcus pneumoniae and Legionella pneumophila determined the affect during such pneumonia. Need for establishment of diagnostic procedure for patients with pneumonia including those caused by atypical agents was revealed.


Industrial monosulfhydryl hydrocarbon compounds-a toxicological and pharmacokinetic profile.
The relevant published toxicological and pharmacokinetic data on the effects of industrial monosulfhydryl hydrocarbon (MSHC) compounds in humans and animals have been reviewed. These compounds are of occupational and environmental exposure concern to humans because they are manufactured in significant quantities, widely used, and insufficiently studied for long-term exposure effects and epidemiology. Both human and animal exposure studies indicate that the inhalation of low molecular weight (C1-C8) compounds at high concentrations may produce severe central nervous system depression and respiratory paralysis. Short-term inhalation at microgram per liter concentrations does not cause any significant toxicity in humans, but the odors do cause environmental concern. Some delayed toxicity and mortality are seen after subchronic exposure in animals. Respiratory organs are significantly damaged by the inhalation of MSHC compounds, whereas some organ pathology is observed in the spleen, liver, and kidneys after oral and intraperitoneal administration. Dermal toxicity is observed for several high molecular weight compounds (> C8). Of all the compounds tested, monosulfhydrylbenzene is most injurious to the eyes, whereas other compounds cause moderate irritation. Major detoxication products of MSHC compounds are carbon dioxide and urinary inorganic sulfate. Methylation and oxidation are the major pathways for detoxication, with the lungs and kidneys being major organ routes of excretion.


The sweet side of the cell cycle.
Cell division (mitosis) and gamete production (meiosis) are fundamental requirements for normal organismal development. The mammalian cell cycle is tightly regulated by different checkpoints ensuring complete and precise chromosomal segregation and duplication. In recent years, researchers have become increasingly interested in understanding how O-GlcNAc regulates the cell cycle. The O-GlcNAc post-translation modification is an O-glycosidic bond of a single β-N-acetylglucosamine sugar to serine/threonine residues of intracellular proteins. This modification is sensitive toward changes in nutrient levels in the cellular environment making O-GlcNAc a nutrient sensor capable of influencing cell growth and proliferation. Numerous studies have established that O-GlcNAcylation is essential in regulating mitosis and meiosis, while loss of O-GlcNAcylation is lethal in growing cells. Moreover, aberrant O-GlcNAcylation is linked with cancer and chromosomal segregation errors. In this review, we will discuss how O-GlcNAc controls different aspects of the cell cycle with a particular emphasis on mitosis and meiosis.


[Counterfeit phosphodiesterase type 5 inhibitors--growing safety risks for public health].
Counterfeit drugs, medical devises and dietary supplements are inherently dangerous and a growing problem. In Europe the growth of the counterfeit medication market is attributable in part to registration of phosphodiesterase type 5 inhibitors (PDE-5) used for the erectile dysfunction. "Viagra, Levitra and Cialis belong to this group. It has been estimated that up to 2.5 million men in Europe are exposed to an illicit sildenafil, suggesting that there may be as many illegal as legal users of sildenafil. In Europe a strong trend is observed towards increasingly professional counterfeits and imitations of Viagra, Cialis and Levitra, with regard to the appearance of tablets, capsules and packaging. The professional presentation will deceive potential consumers into assuming these products are legal, efficacious and safe. Globally, increased obstacles for counterfeiters are necessary to combat pharmaceutical counterfeiting, including fines and penalties. The worldwide nature of the counterfeit problem requires proper coordination between countries to ensure an adequate enforcement. We described the usefulness of the time-of-flight mass spectrometry with the electrospray ionization (LC-ESI-MS-TOF) and the X-ray powder diffraction method (XRPD) for PDE-5 counterfeit screening from the Polish illegal market.


Herpesvirus saimiri DNA in a lymphoid cell line established by in vitro transformation.
A lymphoid T-cell line (H1591) was established by infecting peripheral blood mononuclear cells from a cotton top marmoset with Herpesvirus saimiri OMI. Analysis of these in vitro-immortalized cells revealed nonintegrated, covalently closed circular viral DNA molecules in high multiplicities with substantial rearrangements and large deletions in their L-DNA (unique) regions. One subline, designated H1591 Er, contained circular viral DNA with one stretch of H-DNA (repetitive) and one of L-DNA; the L-DNA segment consisted of a linear fusion of a 53.2-kilobase-pair piece of L-DNA (left half of L-DNA) with a 15.2-kilobase-pair L-DNA fragment from the right end of the L-DNA region. The other subline, H1591 S, contained two short regions of L-DNA, each derived from the extreme ends of virion L-DNA. Both L-DNA regions of H1591 S cells contained inverted repetitions (15.0 +/- 0.2 and 9.1 +/- 4.7 kilobase pairs). The extensive deletions of L-DNA sequences in cell line H1591 indicate that at least 73% of the genetic information in H. saimiri is not required to maintain the persistence of viral DNA and the state of transformation in lymphoid T-cells.


Outcome Selection and Methodological Quality of Major and Minor Shoulder Surgery Studies: A Scoping Review.
Core outcome sets aim to select and standardize the choice of important outcomes reported in clinical trials to encourage more effective data synthesis, increase the reliability of comparing results, and minimize reporting bias. A core outcome set for elective shoulder surgery has yet to be defined, and therefore a systematic assessment of outcomes and methodology is necessary to inform the development of a core outcome set. The purpose of this study was to examine randomized controlled trials (RCTs) of patients having elective major or minor shoulder surgery to (1) identify the outcome domains reported; (2) determine specific outcome measurement tools that were utilized; and (3) assess the work for methodological quality and risk of bias. We conducted a scoping review (a review that identifies the nature and extent of research evidence) to explore the reported outcome domains, outcome tools, and methodological quality from RCTs conducted in shoulder surgery. We considered both major shoulder surgery (defined as arthroplasty, rotator cuff repair, stabilization procedures, biceps tenodesis, or Bankart repairs) and minor shoulder surgery (simple arthroscopy, capsular plication, lateral clavicular excisions, or subacromial decompression). We queried 10 electronic databases for studies published between January 2006 and January 2015. Studies were included if they were prospective, randomized controlled, clinical trials enrolling patients who received an elective shoulder surgical intervention. We extracted data relating to trial characteristics, primary outcomes, tools used to measure these outcomes as well as methodological quality indicators. We assessed indicators of methodological quality by exploring (1) the reproducibility of power analyses; and (2) whether the primary outcomes were powered to minimum clinically important differences. Risk of bias was also assessed with the Jadad score with scores between 0 (very high risk of bias) and 5 (very low risk). Findings were qualitatively analyzed and reported according to systematic and scoping review guidelines. We included 315 studies involving 30,232 patients; 266 studies investigated anesthetic, analgesic, or surgical interventions. Of the 315 studies included, the most common outcome domains evaluated were analgesic (n = 104), functional (n = 87), anesthetic (n = 56), and radiologic (n = 29) outcomes, with temporal patterns noted. Studies of major shoulder surgery most commonly reported functional primary outcome domains, whereas minor shoulder surgery studies most frequently reported analgesic primary outcome domains. There were 85 different primary outcome tools utilized, which included 20 functional, 20 anesthetic, 13 analgesic, and 12 radiologic. A methodological quality assessment revealed that 24% of studies had reproducible power analyses, 13% were powered to minimum clinically important differences, and risk of bias assessment demonstrated a median (interquartile range [range]) Jadad score of 4 (3-5 [1-5]). A wide range of outcome domains and outcome assessment tools are in common use in contemporary trials of patients undergoing elective surgery. Although some diversity is important to allow the assessment of patient populations that may have different goals, the large number of tools in common use may impair the ability of future meta-analyses to pool results effectively or even for systematic reviews to synthesize what is known. The limitations of methodological quality in RCTs may be improved by researchers following standard guidelines and considering the minimum clinically important differences in their trials to be of greater use to clinicians and their patients. Level I, therapeutic study.


Hybrid use of Raman spectroscopy and artificial neural networks to discriminate Mycobacterium bovis BCG and other Mycobacteriales.
Even in the face of the COVID-19 pandemic, Tuberculosis (TB) continues to be a major public health problem and the 2nd biggest infectious cause of death worldwide. There is, therefore, an urgent need to develop effective TB diagnostic methods, which are cheap, portable, sensitive and specific. Raman spectroscopy is a potential spectroscopic technique for this purpose, however, so far, research efforts have focused primarily on the characterisation of Mycobacterium tuberculosis and other Mycobacteria, neglecting bacteria within the microbiome and thus, failing to consider the bigger picture. It is paramount to characterise relevant Mycobacteriales and develop suitable analytical tools to discriminate them from each other. Herein, through the combined use of Raman spectroscopy and the self-optimising Kohonen index network and further multivariate tools, we have successfully undertaken the spectral analysis of Mycobacterium bovis BCG, Corynebacterium glutamicum and Rhodoccocus erythropolis. This has led to development of a useful tool set, which can readily discern spectral differences between these three closely related bacteria as well as generate a unique spectral barcode for each species. Further optimisation and refinement of the developed method will enable its application to other bacteria inhabiting the microbiome and ultimately lead to advanced diagnostic technologies, which can save many lives.


A novel method for in situ synthesis of SERS-active gold nanostars on polydimethylsiloxane film.
A novel one-step in situ synthesis of gold nanostars (AuNSs) on a pre-cured polydimethylsiloxane (PDMS) film is proposed for the fabrication of highly sensitive surface-enhanced Raman scattering (SERS) substrates. Plasmonic activity of the substrates was investigated by collecting SERS maps of 4-mercaptobenzoic acid (4-MBA). The applicability of these flexible substrates is further demonstrated by SERS-based pesticide detection on fruit skin.


Estimating and analyzing the spatial distribution of soil organic carbon in China.
Research on the terrestrial carbon cycle is an important component in the study of global change. Soil organic carbon, as the main part of the terrestrial carbon reservoir, plays an important role in the Earth's carbon cycle. To accurately estimate soil organic carbon storage, its composition and dynamic change must be determined. This presents a challenge to research on the soil carbon cycle, especially in China where the nationwide soil organic carbon reservoir largely remains unknown. This paper reports a research project that attempts to estimate the nationwide soil organic carbon reservoir. Data from 2473 soil profiles from the second national soil survey were collected and GIS technology was employed to quantify the national soil carbon reservoir. The analytical results show that the total amount of soil organic carbon is about 92.4 Pg (Pg = 10(15) g) and that the average carbon density is about 10.53 kg C m(-2). The spatial distribution of soil organic carbon was also analyzed and mapped. This study presents basic data and an analysis method for carbon-cycle studies and also provides scientific support for policy-making efforts to control CO2 emissions in China.


IL-17 inhibits chondrogenic differentiation of human mesenchymal stem cells.
Mesenchymal stem cells (MSCs) can differentiate into cells of mesenchymal lineages, such as osteoblasts and chondrocytes. Here we investigated the effects of IL-17, a key cytokine in chronic inflammation, on chondrogenic differentiation of human MSCs. Human bone marrow MSCs were pellet cultured in chondrogenic induction medium containing TGF-β3. Chondrogenic differentiation was detected by cartilage matrix accumulation and chondrogenic marker gene expression. Over-expression of cartilage matrix and chondrogenic marker genes was noted in chondrogenic cultures, but was inhibited by IL-17 in a dose-dependent manner. Expression and phosphorylation of SOX9, the master transcription factor for chondrogenesis, were induced within 2 days and phosphorylated SOX9 was stably maintained until day 21. IL-17 did not alter total SOX9 expression, but significantly suppressed SOX9 phosphorylation in a dose-dependent manner. At day 7, IL-17 also suppressed the activity of cAMP-dependent protein kinase A (PKA), which is known to phosphorylate SOX9. H89, a selective PKA inhibitor, also suppressed SOX9 phosphorylation, expression of chondrogenic markers and cartilage matrix, and also decreased chondrogenesis. IL-17 inhibited chondrogenesis of human MSCs through the suppression of PKA activity and SOX9 phosphorylation. These results suggest that chondrogenic differentiation of MSCs can be inhibited by a mechanism triggered by IL-17 under chronic inflammation.


An open medical imaging workstation architecture for platform-independent 3-D medical image processing and visualization.
A need for an entirely new medical workstation design was identified to increase the deployment of 3-D medical imaging and multimedia communication. Recent wide acceptance of the Word Wide Web (WWW) as a general communication service within the global network has shown how big the impact of standards and open systems can be. Information is shared among heterogeneous systems and diverse applications on various hardware platforms only by agreeing on a common format for information distribution. For medical image communications, the Digital Imaging and Communication in Medicine (DICOM) standard is possibly anticipating such a role. Logically, the next step is open software: platform-independent tools, which can as easily be transferred and used on multiple platforms. Application of the platform-independent programming language Java enables creation of plug-in tools, which can easily extend the basic system. Performance problems inherent to all interpreter systems can be circumvented by using a hybrid approach. Computationally intensive functions like image processing functions can be integrated into a natively implemented optimized image processing kernel. Plug-in tools implemented in Java can utilize the kernel functions via a Java-wrapper library. This approach is comparable to the implementation of computationally intensive operations in hardware.


The prognostic significance of early treatment response in pediatric relapsed acute myeloid leukemia: results of the international study Relapsed AML 2001/01.
The prognostic significance of early response to treatment has not been reported in relapsed pediatric acute myeloid leukemia. In order to identify an early and easily applicable prognostic factor allowing subsequent treatment modifications, we assessed leukemic blast counts in the bone marrow by morphology on days 15 and 28 after first reinduction in 338 patients of the international Relapsed-AML2001/01 trial. Both day 15 and day 28 status was classified as good (≤20% leukemic blasts) in 77% of patients. The correlation between day 15 and 28 blast percentages was significant, but not strong (Spearman correlation coefficient = 0.49, P<0.001). Survival probability decreased in a stepwise fashion along with rising blast counts at day 28. Patients with bone marrow blast counts at this time-point of ≤5%, 6-10%, 11-20% and >20% had 4-year probabilities of survival of 52%±3% versus 36%±10% versus 21%±9% versus 14%±4%, respectively, P<0.0001; this trend was not seen for day 15 results. Multivariate analysis showed that early treatment response at day 28 had the strongest prognostic significance, superseding even time to relapse (< or ≥12 months). In conclusion, an early response to treatment, measured on day 28, is a strong and independent prognostic factor potentially useful for treatment stratification in pediatric relapsed acute myeloid leukemia. This study was registered with ISRCTN code: 94206677.


Identification of Hybrid Insulin Peptides (HIPs) in Mouse and Human Islets by Mass Spectrometry.
We recently discovered hybrid insulin peptides (HIPs) as a novel class of post-translationally modified peptides in murine-derived beta cell tumors, and we demonstrated that these molecules are autoantigens in type 1 diabetes (T1D). A HIP consists of an insulin fragment linked to another secretory granule peptide via a peptide bond. We verified that autoreactive CD4 T cells in both mouse and human autoimmune diabetes recognize these modified peptides. Here, we use mass spectrometric analyses to confirm the presence of HIPs in both mouse and human pancreatic islets. We also present criteria for the confident identification of these peptides. This work supports the hypothesis that HIPs are autoantigens in human T1D and provides a foundation for future efforts to interrogate this previously unknown component of the beta cell proteome.


Methylation of zebularine investigated using density functional theory calculations.
Deoxyribonucleic acid (DNA) methylation is an epigenetic phenomenon, which adds methyl groups into DNA. This study reveals methylation of a nucleoside antibiotic drug 1-(β-D-ribofuranosyl)-2-pyrimidinone (zebularine or zeb) with respect to its methylated analog, 1-(β-D-ribofuranosyl)-5-methyl-2-pyrimidinone (d5) using density functional theory calculations in valence electronic space. Very similar infrared spectra suggest that zeb and d5 do not differ by types of the chemical bonds, but distinctly different Raman spectra of the nucleoside pair reveal that the impact caused by methylation of zeb can be significant. Further valence orbital-based information details on valence electronic structural changes caused by methylation of zebularine. Frontier orbitals in momentum space and position space of the molecules respond differently to methylation. Based on the additional methyl electron density concentration in d5, orbitals affected by the methyl moiety are classified into primary and secondary contributors. Primary methyl contributions include MO8 (57a), MO18 (47a), and MO37 (28a) of d5, which concentrates on methyl and the base moieties, suggest certain connection to their Frontier orbitals. The primary and secondary methyl affected orbitals provide useful information on chemical bonding mechanism of the methylation in zebularine.


Association between cannabis use and the risk of bladder cancer: results from the California Men's Health Study.
To investigate the association of cannabis use and tobacco smoking on the incidence of bladder cancer within the California Men's Health Study cohort. We evaluated the records of 84,170 participants in a multiethnic cohort of men aged 45-69 years. Information on demographic and lifestyle factors including smoking history and cannabis use was collected using mailed questionnaires between 2002 and 2003. We linked the study data with clinical records including cancer data from electronic health records. Overall 34,000 (41%) cohort members reported cannabis use, 47,092 (57%) reported tobacco use, 22,500 (27%) reported using both, and 23,467 (29%) used neither. Men were followed over an 11-year period and 279 (0.3%) developed incident bladder tumors. Among cannabis users, 89 (0.3%) developed bladder cancer in comparison to 190 (0.4%) men who did not report cannabis use (P < .001). After adjusting for age, race or ethnicity, and body mass index, using tobacco only was associated with an increased risk of bladder cancer (hazard regression [HR], 1.52; 95% confidence interval [CI], 1.12-2.07), whereas cannabis use only was associated with a 45% reduction in bladder cancer incidence (HR, 0.55; 95% CI, 0.31-1.00). Using both cannabis and tobacco was associated with an HR of 1.28 (95% CI, 0.91-1.80). Although a cause and effect relationship has not been established, cannabis use may be inversely associated with bladder cancer risk in this population.


Two-Year Evaluation of Osteochondral Repair with a Novel Biphasic Graft Saturated in Bone Marrow in an Equine Model.
Objective To evaluate a biphasic cartilage repair device (CRD) for feasibility of arthroscopic implantation, safety, biocompatibility, and efficacy for long-term repair of large osteochondral defects. Methods The CRD was press-fit into defects (10 mm diameter, 10 mm deep) created in the femoral trochlea of 12 horses. In the contralateral limb, 10 mm diameter full-thickness chondral defects were treated with microfracture (MFX). Radiographs were obtained pre- and postoperatively, and at 4, 12, and 24 months. Repeat arthroscopy was performed at 4 and 12 months. Gross assessment, histology, mechanical testing, and magnetic resonance imaging (MRI) were performed at 24 months. Results The CRD was easily placed arthroscopically. There was no evidence of joint infection, inflammation, or degeneration. CRD-treated defects had significantly more sclerosis compared to MFX early ( P = 0.0006), but was not different at 24 months. CRD had better arthroscopic scores at 4 months compared to MFX ( P = 0.0069). At 24 months, there was no difference in repair tissue on histology or mechanical testing. Based on MRI, CRD repair tissue had less proteoglycan (deep P = 0.027, superficial P = 0.015) and less organized collagen (deep P = 0.028) compared to MFX. Cartilage surrounding MFX defects had more fissures compared to CRD. Conclusion The repair tissue formed after CRD treatment of a large osteochondral lesion is fibrocartilage similar to that formed in simple chondral defects treated with MFX. The CRD can be easily placed arthroscopically, is safe, and biocompatible for 24 months. The CRD results in improved early arthroscopic repair scores and may limit fissure formation in adjacent cartilage.


Functional and structural characterization of the zebrafish Na+-sulfate cotransporter 1 (NaS1) cDNA and gene (slc13a1).
Sulfate plays an essential role during growth, development and cellular metabolism. In this study, we characterized the function and structure of the zebrafish (Danio rerio) Na+-sulfate cotransporter 1 (NaS1) cDNA and gene (slc13a1). Zebrafish NaS1 encodes a protein of 583 amino acids with 13 putative transmembrane domains. Expression of zebrafish NaS1 protein in Xenopus oocytes led to Na+-sulfate cotransport, which was significantly inhibited by thiosulfate, selenate, molybdate, and tungstate. Zebrafish NaS1 transport kinetics were: V(max) = 1,731.670 +/- 92.853 pmol sulfate/oocyte.hour and K(m) = 1.414 +/- 0.275 mM for sulfate and V(max) = 307.016 +/- 32.992 pmol sulfate/oocyte x hour, K(m) = 24.582 +/- 4.547 mM and n (Hill coefficient) = 1.624 +/- 0.354 for sodium. Zebrafish NaS1 mRNA is developmentally expressed in embryos from day 1 postfertilization and in the intestine, kidney, brain, and eye of adult zebrafish. The zebrafish NaS1 gene slc13a1 contains 15 exons spanning 8,716 bp. Characterization of the zebrafish NaS1 contributes to a greater understanding of sulfate transporters in a well-defined genetic model and will allow the elucidation of evolutionary and functional relationships among vertebrate sulfate transporters.


Dynamic-clamp analysis of wild-type human Nav1.7 and erythromelalgia mutant channel L858H.
The link between sodium channel Nav1.7 and pain has been strengthened by identification of gain-of-function mutations in patients with inherited erythromelalgia (IEM), a genetic model of neuropathic pain in humans. A firm mechanistic link to nociceptor dysfunction has been precluded because assessments of the effect of the mutations on nociceptor function have thus far depended on electrophysiological recordings from dorsal root ganglia (DRG) neurons transfected with wild-type (WT) or mutant Nav1.7 channels, which do not permit accurate calibration of the level of Nav1.7 channel expression. Here, we report an analysis of the function of WT Nav1.7 and IEM L858H mutation within small DRG neurons using dynamic-clamp. We describe the functional relationship between current threshold for action potential generation and the level of WT Nav1.7 conductance in primary nociceptive neurons and demonstrate the basis for hyperexcitability at physiologically relevant levels of L858H channel conductance. We demonstrate that the L858H mutation, when modeled using dynamic-clamp at physiological levels within DRG neurons, produces a dramatically enhanced persistent current, resulting in 27-fold amplification of net sodium influx during subthreshold depolarizations and even greater amplification during interspike intervals, which provide a mechanistic basis for reduced current threshold and enhanced action potential firing probability. These results show, for the first time, a linear correlation between the level of Nav1.7 conductance and current threshold in DRG neurons. Our observations demonstrate changes in sodium influx that provide a mechanistic link between the altered biophysical properties of a mutant Nav1.7 channel and nociceptor hyperexcitability underlying the pain phenotype in IEM.


The level of erbB2 expression predicts sensitivity to the cytotoxic effects of an intracellular anti-erbB2 sFv.
We have previously demonstrated that an intracellular antibody (sFv) directed against erbB2 can achieve a specific cytotoxicity in erbB2 overexpressing cancer cells of varying histogenesis. In order to further delineate the mechanistic basis of the induced apoptosis, transient and stable cotransfections were performed. Transient cotransfection of erbB2 mutant and chimeric molecules demonstrated that the cytoplasmic domain of erbB2, or the homologous cytoplasmic domain of the epidermal growth factor receptor, is required for apoptosis induction. These results were confirmed in assays utilizing differential derivation of stable clones. To examine the effects of varying ratios of the anti-erbB2 sFv and its target erbB2 we performed additional cotransfection experiments in erbB2 negative target cells. When erbB2 levels are held constant, observed cytotoxicity is proportional to the amount of sFv added. In addition, when sFv levels are held constant, increasing levels of cotransfected erbB2 can overcome the apoptotic response. These results indicate that a minimal threshold level of the sFv and its target are required to induce cytotoxicity. To examine this phenomenon in an erbB2 positive cell line, SKOV3 ovarian carcinoma cells were utilized to derive a stable clone expressing low levels of sFv. When this cell line was compared to the parental SKOV3 cell line, it was shown that less exogenous sFv was needed to induce cytotoxicity in the clone already expressing low levels of sFv, indicating that endogenous and exogenous levels of sFv are additive. In summary, the results presented here indicate that the carboxy-terminus of the intracellular domain of the erbB2 molecule is involved in the induction of apoptosis. Furthermore, the expression levels of the sFv and its target protein need to overcome a threshold level in order to achieve a cytotoxic response.