Chicken egg fetal liver DNA and histopathologic effects of structurally diverse carcinogens and non-carcinogens.
Chicken egg fetal livers were evaluated for histopathological changes produced by four genotoxic hepatocarcinogens: 2-acetylaminofluorene (AAF), aflatoxin B1 (AFB1), benzo[a]pyrene (BaP), diethylnitrosamine (DEN); four structurally related non- or weakly- carcinogenic comparators: fluorene (FLU), aflatoxin B2 (AFB2), benzo[e]pyrene (BeP), N-nitrosodiethanolamine (NDELA); two epigenetic hepatocarcinogens: clofibric acid (CFA), phenobarbital (PB); and the non-carcinogen, D-mannitol (MAN). CFA, PB and MAN were also assessed for formation of DNA adducts using the 32P nucleotide postlabeling (NPL) assay and for DNA breaks using the comet assay. CFA was also assessed in enhanced comet assay for oxidative DNA damage induction. Eggs were dosed on days 9- 11 of incubation. For genotoxicity evaluation, livers were collected 3h after the last dose. Liver qualitative histopathology assessment was performed on days 12 and 18 of incubation. CFA was negative for DNA adducts but yielded clear evidence of DNA breaks due to oxidative stress. PB and MAN produced no DNA adducts or breaks. Liver to body weight ratios were not affected in most groups, but were decreased in DEN groups, and increased after PB dosing. Livers from control groups, FLU, AFB2, BeP, NDELA, CFA, and MAN groups, displayed a typical hepatocellular trabecular pattern at both time points. In contrast, the four genotoxic carcinogens induced time- and dose- related interference with fetal liver cell processes of proliferation, migration and differentiation, leading to hepatocellular and cholangiocellular pleomorphic dysplasia and re-(de-) differentiation with distortion of the trabecular pattern. In addition, dosing with the high dose of DEN produced gallbladder agenesis. PB induced hepatocellular hypertrophy, interference with migration, expressed as distortion of the trabecular pattern, and a moderate cholangiocellular dysplasia. In summary, histopathological analysis of chicken fetal livers revealed developmental anomalies, as well as genotoxicity-induced and, in the case of PB, adaptive morphological changes. Thus, the model provides histopathological outcomes of molecular effects.


Radiopacity of zirconia-based all-ceramic crown systems.
The aim of this study was to evaluate the radiopacity of the core and veneer ceramics in four zirconia-based crown systems: In-Ceram Zirconia, In-Ceram YZ, Procera Zirconia, and Cercon. Two-millimeter-thick restoration slices were prepared and digitally radiographed alongside an aluminum stepwedge. The grayscale data were converted into millimeters of aluminum by an image analysis program. The two-way analysis of variance test detected significant differences with respect to all-ceramic system, layer ceramic, and the interaction of the two factors (P = .0001). All materials presented radiopacity values above the minimum recommended by the International Organization for Standardization.


The effect of tone-vocoding on spatial release from masking for old, hearing-impaired listeners.
Old, hearing-impaired listeners generally benefit little from lateral separation of multiple talkers when listening to one of them. This study aimed to determine how spatial release from masking (SRM) in such listeners is affected when the interaural time differences (ITDs) in the temporal fine structure (TFS) are manipulated by tone-vocoding (TVC) at the ears by a master hearing aid system. Word recall was compared, with and without TVC, when target and masker sentences from a closed set were played simultaneously from the front loudspeaker (co-located) and when the maskers were played 45° to the left and right of the listener (separated). For 20 hearing-impaired listeners aged 64 to 86, SRM was 3.7 dB smaller with TVC than without TVC. This difference in SRM correlated with mean audiometric thresholds below 1.5 kHz, even when monaural TFS sensitivity (discrimination of frequency-shifts in identically filtered complexes) was partialed out, suggesting that low-frequency audiometric thresholds may be a good indicator of candidacy for hearing aids that preserve ITDs. The TVC difference in SRM was not correlated with age, pure-tone ITD thresholds, nor fundamental frequency difference limens, and only with monaural TFS sensitivity before control for low-frequency audiometric thresholds.


Web-based learning in undergraduate medical education: development and assessment of an online course on experimental surgery.
In order to increase the number of practical and discussion classes offered to students in the traditional-curriculum scenario, while decreasing the lecture-based ones and to create an online community to share knowledge on surgery, we developed and assessed the first online course for undergraduate medical students on experimental surgery at the Federal University of Sao Paulo-UNIFESP, Brazil. The purposes of the present study are: describe and discuss the process and the lessons learned involved in developing an undergraduate web-based course and analyze the students' attitude towards this educational environment. A group of medical students was taught online during 5 weeks on the theory of experimental surgery through video quizzes, required readings, collaborative activities using discussion board and asynchronous communication. The students' knowledge gain, their web session variables and the results of the course evaluation were used to support our study. The students have significantly improved their knowledge on experimental surgery after the course. Among factors in the online course that could possibly have contributed to this gain, the interactive activities (video quizzes), key element in our online material, seemed to be promising for candidates. The evaluation results demonstrated high levels of course functionality, effectiveness of its online content and acceptance among medical students. This study indicated that a web-based course for undergraduate students may be successfully developed and implemented in medical settings and the students seem to be quite supportive. We encourage undergraduate medical learning strategies involving the Web.


Functional study of elafin cleaved by Pseudomonas aeruginosa metalloproteinases.
Elafin is a 6-kDa innate immune protein present at several epithelial surfaces including the pulmonary epithelium. It is a canonical protease inhibitor of two neutrophil serine proteases [neutrophil elastase (NE) and proteinase 3] with the capacity to covalently bind extracellular matrix proteins by transglutamination. In addition to these properties, elafin also possesses antimicrobial and immunomodulatory activities. The aim of the present study was to investigate the effect of Pseudomonas aeruginosa proteases on elafin function. We found that P. aeruginosa PAO1-conditioned medium and two purified Pseudomonas metalloproteases, pseudolysin (elastase) and aeruginolysin (alkaline protease), are able to cleave recombinant elafin. Pseudolysin was shown to inactivate the anti-NE activity of elafin by cleaving its protease-binding loop. Interestingly, antibacterial properties of elafin against PAO1 were found to be unaffected after pseudolysin treatment. In contrast to pseudolysin, aeruginolysin failed to inactivate the inhibitory properties of elafin against NE. Aeruginolysin cleaves elafin at the amino-terminal Lys6-Gly7 peptide bond, resulting in a decreased ability to covalently bind purified fibronectin following transglutaminase activity. In conclusion, this study provides evidence that elafin is susceptible to proteolytic cleavage at alternative sites by P. aeruginosa metalloproteinases, which can affect different biological functions of elafin.


Risk assessment of hepatocellular carcinoma in chronic hepatitis C patients by transient elastography.
The degree of liver fibrosis is the strongest indicator of risk for hepatocellular carcinoma (HCC) development. Recently developed transient elastography (Fibroscan, Echosens, France) noninvasively measures liver stiffness, and the correlation between the stiffness and liver fibrosis stage has been validated. In this cross-sectional study, we investigated the relationship between liver stiffness and HCC presence. Liver stiffness was measured in chronic hepatitis C patients (85 with HCC and 180 without) by transient elastography. Multivariate logistic regression was applied to assess the association with HCC presence. We computed the receiver operating characteristics (ROC) curves concerning the prediction of HCC presence and compared the areas under ROC curve (AUROC). We also calculated stratum-specific likelihood ratios (SSLR). Multivariate analysis showed that HCC presence was significantly associated with liver stiffness (P<0.0001) along with age, male, and alpha-fetoprotein concentration. AUROC was 0.805, 0.741, 0.714, 0.673, 0.670, and 0.654 for liver stiffness, alpha-fetoprotein, albumin, prothrombin activity, AST-platelet ratio index, and platelet count, respectively. Other parameters showed smaller AUROC. SSLR for HCC presence by liver stiffness was 0.22 (95% confidence interval: 0.11-0.42) in <10 kPa, 0.73 (0.39 to 1.39) in 10.1 to 15 kPa, 1.30 (0.80 to 2.12) in 15.1 to 25 kPa, and 5.0 (2.96 to 8.47) in >25 kPa. Liver stiffness measured by transient elastography is useful in demarcating chronic hepatitis C patients at a high risk for HCC, who require frequent check-up by imaging examinations.


Structures of respiratory syncytial virus nucleocapsid protein from two crystal forms: details of potential packing interactions in the native helical form.
Respiratory syncytial virus (RSV) is a frequent cause of respiratory illness in infants, but there is currently no vaccine nor effective drug treatment against this virus. The RSV RNA genome is encapsidated and protected by a nucleocapsid protein; this RNA-nucleocapsid complex serves as a template for viral replication. Interest in the nucleocapsid protein has increased owing to its recent identification as the target site for novel anti-RSV compounds. The crystal structure of human respiratory syncytial virus nucleocapsid (HRSVN) was determined to 3.6 Å resolution from two crystal forms belonging to space groups P2(1)2(1)2(1) and P1, with one and four decameric rings per asymmetric unit, respectively. In contrast to a previous structure of HRSVN, the addition of phosphoprotein was not required to obtain diffraction-quality crystals. The HRSVN structures reported here, although similar to the recently published structure, present different molecular packing which may have some biological implications. The positions of the monomers are slightly shifted in the decamer, confirming the adaptability of the ring structure. The details of the inter-ring contacts in one crystal form revealed here suggest a basis for helical packing and that the stabilization of native HRSVN is via mainly ionic interactions.


Does vertebral level of pedicle subtraction osteotomy correlate with degree of spinopelvic parameter correction?
Pedicle subtraction osteotomy (PSO) is a spinal realignment technique that may be used to correct sagittal spinal imbalance. Theoretically, the level and degree of resection via a PSO should impact the degree of sagittal plane correction in the setting of deformity. However, the quantitative effect of PSO level and focal angular change on postoperative spinopelvic parameters has not been well described. The purpose of this study is to analyze the relationship between the level/degree of PSO and changes in global sagittal balance and spinopelvic parameters. In this multicenter retrospective study, 70 patients (54 women and 16 men) underwent lumbar PSO surgery for spinal imbalance. Preoperative and postoperative free-standing sagittal radiographs were obtained and analyzed by regional curves (lumbar, thoracic, and thoracolumbar), pelvic parameters (pelvic incidence and pelvic tilt [PT]) and global balance (sagittal vertical axis [SVA] and T-1 spinopelvic inclination). Correlations between PSO parameters (level and degree of change in angle between the 2 adjacent vertebrae) and spinopelvic measurements were analyzed. Pedicle subtraction osteotomy distribution by level and degree of correction was as follows: L-1 (6 patients, 24°), L-2 (15 patients, 24°), L-3 (29 patients, 25°), and L-4 (20 patients, 22°). There was no significant difference in the focal correction achieved by PSO by level. All patients demonstrated changes in preoperative to postoperative parameters including increased lumbar lordosis (from 20° to 49°, p < 0.001), increased thoracic kyphosis (from 30° to 38°, p < 0.001), decreased SVA and T-1 spinopelvic inclination (from 122 to 34 mm, p < 0.001 and from +3° to -4°, p < 0.001, respectively), and decreased PT (from 31° to 23°, p < 0.001). More caudal PSO was correlated with greater PT reduction (r = -0.410, p < 0.05). No correlation was found between SVA correction and PSO location. The PSO degree was correlated with change in thoracic kyphosis (r = -0.474, p < 0.001), lumbar lordosis (r = 0.667, p < 0.001), sacral slope (r = 0.426, p < 0.001), and PT (r = -0.358, p < 0.005). The degree of PSO resection correlates more with spinopelvic parameters (lumbar lordosis, thoracic kyphosis, PT, and sacral slope) than PSO level. More importantly, PSO level impacts postoperative PT correction but not SVA.


Polychlorinated biphenyl (PCB) load, lipid reserves and biotransformation activity in migrating Atlantic salmon from River Mörrum, Sweden.
Atlantic salmon accumulate high levels of contaminants such as polychlorinated biphenyls (PCBs) in their lipids during the adult growth phase spent at sea. The lipids are later utilized during migration for swimming and biological adaptations. We hypothesize that migrating salmons' biotransformation processes are affected by the high levels of built-up PCBs compared to salmon that in a pre-migrational stage. For these analyses we sampled adult Atlantic salmon during migration in the Swedish River Mörrum and measured the 21 most common PCB congeners ( summation operatorPCB) and lipid levels in muscle tissue, aryl hydrocarbon receptor (AHR2) and cytochrome P4501A1 (CYP1A1) transcript levels as well as ethoxyresorufin-O-deethylase activity (EROD) in liver. We also determined which AHR2 genotypes the salmon carried. We show that EROD activity is correlated to CYP1A1 level but not to summation operatorPCB concentration. summation operatorPCB concentration does not predict levels of neither the AHR2 nor CYP1A1 genes. We find no associations between specific AHR2 transcription levels and AHR2 genotypes or a correlation between AHR2 and CYP1A1 transcription levels, which is in direct contrast to pre-migrational adult salmon from the Baltic Sea. When we compare River Mörrum to salmon we have previously sampled in the Baltic Sea we show that migrating salmon have significantly lower lipid levels in their muscles; higher muscle concentrations of summation operatorPCB on a lipid basis; and significantly lower CYP1A1 and EROD levels compared to salmon from the Baltic Sea. Also, transcript levels of three out of four AHR2 genes are significantly different. In conclusion, migrating Swedish Atlantic salmon carry higher concentrations of PCBs in their lipids compared to salmon in the Baltic Sea, but have lower activation of biotransformation genes and enzymes. Our results indicate that accumulated pollutants from the Baltic Sea are deactivated inside the migrating salmon's lipid tissues and increase in concentration when migration is initiated thereby limiting their impact on biotransformation processes.


Hypothyroidism and myocardial failure in two Great Danes.
Two Great Danes were evaluated for signs of congestive heart failure. Dilated cardiomyopathy was diagnosed in each dog. Both dogs were diagnosed concurrently with primary hypothyroidism. Following appropriate supplementation with levothyroxine, both dogs showed significant improvement in myocardial contractility, as demonstrated by increased fractional shortening and reductions in left atrial size, and left ventricular end-systolic and diastolic diameters. These observations suggest that hypothyroidism may lead to myocardial failure in Great Danes that is partially or fully reversible following thyroid hormone replacement therapy, and that hypothyroidism should be considered in all Great Danes presenting with systolic failure.


A novel C53/LZAP-interacting protein regulates stability of C53/LZAP and DDRGK domain-containing Protein 1 (DDRGK1) and modulates NF-kappaB signaling.
C53/LZAP (also named as Cdk5rap3) is a putative tumor suppressor that plays important roles in multiple cell signaling pathways, including DNA damage response and NF-kappaB signaling. Yet how its function is regulated remains largely unclear. Here we report the isolation and characterization of two novel C53/LZAP-interacting proteins, RCAD (Regulator of C53/LZAP and DDRGK1) and DDRGK1 (DDRGK domain-containing protein 1). Our co-immunoprecipitation assays confirmed their interactions, while gel filtration assay indicated that C53/LZAP and RCAD may form a large protein complex. Intriguingly, we found that RCAD knockdown led to dramatic reduction of C53/LZAP and DDRGK1 proteins. We also found that C53/LZAP and DDRGK1 became more susceptible to the proteasome-mediated degradation in RCAD knockdown cells, whereas their ubiquitination was significantly attenuated by RCAD overexpression. In addition, we found that RCAD, like C53/LZAP, also plays an important role in regulation of NF-kappaB signaling and cell invasion. Taken together, our findings strongly suggest that RCAD is a novel regulator of C53/LZAP tumor suppressor and NF-kappaB signaling.


Fungal radiation in the Cape Floristic Region: an analysis based on Gondwanamyces and Ophiostoma.
The Cape Floristic Region (CFR) displays high levels of plant diversity and endemism, and has received focused botanical systematic attention. In contrast, fungal diversity patterns and co-evolutionary processes in this region have barely been investigated. Here we reconstruct molecular phylogenies using the ITS and beta-tubulin gene regions of the ophiostomatoid fungi Gondwanamyces and Ophiostoma associated with southern African Protea species. Results indicate that they evolved in close association with Protea. In contrast to Protea, Ophiostoma species migrated to the CFR from tropical and subtropical Africa, where they underwent subsequent radiation. In both Gondwanamyces and Ophiostoma vector arthropods probably facilitated long-distance migration and shorter-distance dispersal. Although ecological parameters shaped most associations between ophiostomatoid fungi and Protea, there is congruence between fungal-host-associations and the systematic classification of Protea. These results confirm that the entire biotic environment must be considered in order to understand diversity and evolution in the CFR as a whole.


Upper respiratory tract colonization with Streptococcus pneumoniae in adults.
Most of the current evidence regarding pneumococcal upper respiratory colonization in adults suggests that despite high disease burden, carriage prevalence is low. Contemporary studies on adult pneumococcal colonization have largely followed the pediatric approach by which samples are obtained mostly from the nasopharynx and bacterial detection is evaluated by routine culture alone. Recent evidence suggests that the 'pediatric approach' may be insufficient in adults and pneumococcal detection in this population may be improved by longitudinal studies that include samples from additional respiratory sites combined with more extensive laboratory testing. In this article, relevant literature published in peer review journals on adult pneumococcal colonization, epidemiology, detection methods, and recommendations were reviewed. Respiratory carriage of Streptococcus pneumoniae has been underestimated in adults. Contemporary pneumococcal carriage studies in adults that collect samples from alternative respiratory sites such as the oropharynx, saliva, or nasal wash; are culture-enriched for pneumococcus; and use molecular diagnostic methods designed to target two pneumococcal DNA sequences should enhance pneumococcal detection in the adult respiratory tract. This finding may have implications for the interpretation of dynamics of pneumococcal transmission and vaccination.


Bioinspired Star-Shaped Poly(l-lysine) Polypeptides: Efficient Polymeric Nanocarriers for the Delivery of DNA to Mesenchymal Stem Cells.
The field of tissue engineering is increasingly recognizing that gene therapy can be employed for modulating in vivo cellular response thereby guiding tissue regeneration. However, the field lacks a versatile and biocompatible gene delivery platform capable of efficiently delivering transgenes to mesenchymal stem cells (MSCs), a cell type often refractory to transfection. Herein, we describe the extensive and systematic exploration of three architectural variations of star-shaped poly(l-lysine) polypeptide (star-PLL) with varying number and length of poly(l-lysine) arms as potential nonviral gene delivery vectors for MSCs. We demonstrate that star-PLL vectors are capable of self-assembling with pDNA to form stable, cationic nanomedicines. Utilizing high content screening, live cell imaging, and mechanistic uptake studies we confirm the intracellular delivery of pDNA by star-PLLs to MSCs is a rapid process, which likely proceeds via a clathrin-independent mechanism. We identify a star-PLL composition with 64 poly(l-lysine) arms and five l-lysine subunits per arm as a particularly efficient vector that is capable of delivering both reporter genes and the therapeutic transgenes bone morphogenetic protein-2 and vascular endothelial growth factor to MSCs. This composition facilitated a 1000-fold increase in transgene expression in MSCs compared to its linear analogue, linear poly(l-lysine). Furthermore, it demonstrated comparable transgene expression to the widely used vector polyethylenimine using a lower pDNA dose with significantly less cytotoxicity. Overall, this study illustrates the ability of the star-PLL vectors to facilitate efficient, nontoxic nucleic acid delivery to MSCs thereby functioning as an innovative nanomedicine platform for tissue engineering applications.


The signaling mechanisms of syndecan heparan sulfate proteoglycans.
Syndecans are membrane proteins controlling cell proliferation, differentiation, adhesion, and migration. Their extracellular domains bear versatile heparan sulfate chains that provide structural determinants for syndecans to function as coreceptors or activators for molecules like growth factors and constituents of the matrix. Syndecans also signal via their protein cores and their conserved transmembrane and cytoplasmic domains. The direct interactions and signaling cascades they support are becoming better characterized. These interactions are regulated by phosphorylation, induced clustering and shedding of the syndecan extracellular domain. Moreover evidence is emerging that syndecans concentrate in unconventional lipid domains and might govern novel vesicular trafficking pathways. Here we focus on recent findings that refine our understanding of the complex structure-function relationships of these cellular effectors.


Tumor Microenvironment and Hydrogel-Based 3D Cancer Models for In Vitro Testing Immunotherapies.
In recent years, immunotherapy has emerged as a promising novel therapeutic strategy for cancer treatment. In a relevant percentage of patients, however, clinical benefits are lower than expected, pushing researchers to deeply analyze the immune responses against tumors and find more reliable and efficient tools to predict the individual response to therapy. Novel tissue engineering strategies can be adopted to realize in vitro fully humanized matrix-based models, as a compromise between standard two-dimensional (2D) cell cultures and animal tests, which are costly and hardly usable in personalized medicine. In this review, we describe the main mechanisms allowing cancer cells to escape the immune surveillance, which may play a significant role in the failure of immunotherapies. In particular, we discuss the role of the tumor microenvironment (TME) in the establishment of a milieu that greatly favors cancer malignant progression and impact on the interactions with immune cells. Then, we present an overview of the recent in vitro engineered preclinical three-dimensional (3D) models that have been adopted to resemble the interplays between cancer and immune cells and for testing current therapies and immunotherapeutic approaches. Specifically, we focus on 3D hydrogel-based tools based on different types of polymers, discussing the suitability of each of them in reproducing the TME key features based on their intrinsic or tunable characteristics. Finally, we introduce the possibility to combine the 3D models with technological fluid dynamics platforms, reproducing the dynamic complex interactions between tumor cells and immune effectors migrated in situ via the systemic circulation, pointing out the challenges that still have to be overcome for setting more predictive preclinical assays.


Chromosomal sites of integration of simian virus 40 DNA sequences mapped by in situ hybridization in two transformed hybrid cell lines.
Simian virus 40 DNA sequences, integrated on human chromosome 7 in two transformed human-mouse hybrid somatic cell lines, were mapped to a specific chromosomal locus by in situ hybridization. To detect the integrated viral DNA by in situ hybridization, we increased the sensitivity of the technique by using as a probe 125I-labeled simian virus 40 cRNA (2 X 10(9) to 3 X 10(9) dpm/micrograms) prepared by in vitro transcription of simian virus 40 DNA with high-specific-radioactivity [125I]CTP. Although the viral nucleic acid was shown by blot hybridization in the two cell lines to be integrated in different restriction fragments, it was shown by in situ hybridization in the two cell lines to map to the same position, q31, on the long arm of human chromosome 7. The viral DNA integration sites were localized with a precision of +/- 1 silver grain diameter, equivalent to about 6.2 X 10(7) nucleotide pairs in the human genome. The procedures we describe may be adapted for localization of any gene on diploid chromosomes that can be cloned in a recombinant DNA vector.


Estradiol promotes EMT in endometriosis via MALAT1/miR200s sponge function.
Endometriosis is an estrogen-dependent benign gynecological disease that shares some common features of malignancy. Epithelial-mesenchymal transition (EMT) has been recognized as a core mechanism of endometriosis. MALAT1 is widely known as EMT promoter, while miR200 family members (miR200s) are considered as EMT inhibitors. Previous studies have reported that MALAT1 upregulation and miR200s downregulation are observed in endometriosis. MiR200c has been regarded as the strongest member of miR200s to interact with MALAT1. However, whether MALAT1/miR200c regulates EMT remains largely unclear. In this study, the roles of miR200s and MALAT1 in ectopic endometrium were investigated. Additionally, the effects of E2 on EMT and MALAT1/miR200s were examined in both EECs and Ishikawa cells. Notably, E2 could upregulate MALAT1 and downregulate miR200s expression levels and induce EMT in EECs and Ishikawa cells. PHTPP, an ERβ antagonist, could reverse the effect of E2. Overexpression of miR200c and knockdown of MALAT1 significantly inhibited E2-mediated EMT, suggesting that both miR200c and MALAT1 are involved in the E2-induced EMT process in endometriosis. In addition, a reciprocal inhibition was found between miR200s and MALAT1. Therefore, the role of MALAT1/miR200c in EMT is influenced by the presence of estrogen during endometriosis development.


Structural characterization of the RNase E S1 domain and identification of its oligonucleotide-binding and dimerization interfaces.
S1 domains occur in four of the major enzymes of mRNA decay in Escherichia coli: RNase E, PNPase, RNase II, and RNase G. Here, we report the structure of the S1 domain of RNase E, determined by both X-ray crystallography and NMR spectroscopy. The RNase E S1 domain adopts an OB-fold, very similar to that found with PNPase and the major cold shock proteins, in which flexible loops are appended to a well-ordered five-stranded beta-barrel core. Within the crystal lattice, the protein forms a dimer stabilized primarily by intermolecular hydrophobic packing. Consistent with this observation, light-scattering, chemical crosslinking, and NMR spectroscopic measurements confirm that the isolated RNase E S1 domain undergoes a specific monomer-dimer equilibrium in solution with a K(D) value in the millimolar range. The substitution of glycine 66 with serine dramatically destabilizes the folded structure of this domain, thereby providing an explanation for the temperature-sensitive phenotype associated with this mutation in full-length RNase E. Based on amide chemical shift perturbation mapping, the binding surface for a single-stranded DNA dodecamer (K(D)=160(+/-40)microM) was identified as a groove of positive electrostatic potential containing several exposed aromatic side-chains. This surface, which corresponds to the conserved ligand-binding cleft found in numerous OB-fold proteins, lies distal to the dimerization interface, such that two independent oligonucleotide-binding sites can exist in the dimeric form of the RNase E S1 domain. Based on these data, we propose that the S1 domain serves a dual role of dimerization to aid in the formation of the tetrameric quaternary structure of RNase E as described by Callaghan et al. in 2003 and of substrate binding to facilitate RNA hydrolysis by the adjacent catalytic domains within this multimeric enzyme.


Xenopus oocyte K+ current. II. Adenylyl cyclase-linked receptors on follicle cells.
Xenopus ovarian follicles consist of single large oocytes surrounded by a layer of small follicle cells that are coupled to the oocyte by gap junctions. Hyperpolarizing K+ currents can be detected in the oocytes of follicles stimulated with adenosine, isoproterenol, follicle-stimulating hormone (FSH), or microinjected adenosine 3',5'-cyclic monophosphate (cAMP). We show that cAMP accumulation can be detected in follicles incubated with the adenosine agonist 5'-N-ethylcarboxamidoadenosine (NECA), isoproterenol, or FSH, but only if forskolin and a phosphodiesterase inhibitor are also added. Treatment of follicles with collagenase has been reported to reduce, but usually not to eliminate, cAMP-activated K+ currents. In this study we show that collagenase treatment alone does not completely remove follicle cells or receptor-mediated cAMP accumulation measured in follicles. cAMP accumulation and cAMP-dependent K+ currents are both eliminated when the follicle cells are completely removed by a technique involving treatment of follicles with collagenase and hypertonic saline. Oocytes completely stripped of follicle cells fail to accumulate cAMP in response to receptor agonists and forskolin. Isolated follicle cells derived from single follicles (but without the oocyte present) accumulate cAMP in response to these drugs to an extent equivalent to the response seen in single intact follicles. Adenylyl cyclase-linked receptors of Xenopus follicles thus appear to be located exclusively on follicle cells. The data suggest that cAMP-dependent K+ currents, although measured in oocytes, may be generated in follicle cells which communicate with oocytes. Another possibility is that a high resting K+ conductance in follicle cells is communicated to oocytes via cAMP-sensitive gap junctions.


The human genus.
A general problem in biology is how to incorporate information about evolutionary history and adaptation into taxonomy. The problem is exemplified in attempts to define our own genus, Homo. Here conventional criteria for allocating fossil species to Homo are reviewed and are found to be either inappropriate or inoperable. We present a revised definition, based on verifiable criteria, for Homo and conclude that two species, Homo habilis and Homo rudolfensis, do not belong in the genus. The earliest taxon to satisfy the criteria is Homo ergaster, or early African Homo erectus, which currently appears in the fossil record at about 1.9 million years ago.


Modulation of preference for abstract stimuli following competence-based social status primes.
In the present study, we measured whether competence-related high and low social status attributed to two unknown individuals affects participants' implicit reactivity to abstract stimuli associated to the identity of the same individuals. During a status-inducing procedure, participants were asked to play an interactive game with two (fake) players coded as high vs low status based on their game competence. Before and after the game, a modified version of the Affective Misattribution Procedure (AMP) was administered in which the players' faces were used as primes. The evaluation target, as is typical to AMP, was a Chinese ideogram. There were two different presentation timings for the prime image: 75 ms and 17 ms. After the status-inducing procedure, the evaluation targets preceded by the high-status prime (i.e. best player's face) were rated as more pleasant than those preceded by the low-status prime (i.e. worst player's face). This effect was only found, however, for the 75 ms lasting prime. Moreover, explicit ratings of the primes showed that the high-status player was rated as more intelligent, competent and dominant than the low status one. These results indicate that implicit preference and explicit evaluation of unacquainted individuals are rapidly modulated by competence-based social status attribution, thus hinting at the plastic nature of social categorization and, relatedly, the malleability of visual preference.


Comprehensive Ex Vivo Transposon Mutagenesis Identifies Genes That Promote Growth Factor Independence and Leukemogenesis.
Aberrant signaling through cytokine receptors and their downstream signaling pathways is a major oncogenic mechanism underlying hematopoietic malignancies. To better understand how these pathways become pathologically activated and to potentially identify new drivers of hematopoietic cancers, we developed a high-throughput functional screening approach using ex vivo mutagenesis with the Sleeping Beauty transposon. We analyzed over 1,100 transposon-mutagenized pools of Ba/F3 cells, an IL3-dependent pro-B-cell line, which acquired cytokine independence and tumor-forming ability. Recurrent transposon insertions could be mapped to genes in the JAK/STAT and MAPK pathways, confirming the ability of this strategy to identify known oncogenic components of cytokine signaling pathways. In addition, recurrent insertions were identified in a large set of genes that have been found to be mutated in leukemia or associated with survival, but were not previously linked to the JAK/STAT or MAPK pathways nor shown to functionally contribute to leukemogenesis. Forced expression of these novel genes resulted in IL3-independent growth in vitro and tumorigenesis in vivo, validating this mutagenesis-based approach for identifying new genes that promote cytokine signaling and leukemogenesis. Therefore, our findings provide a broadly applicable approach for classifying functionally relevant genes in diverse malignancies and offer new insights into the impact of cytokine signaling on leukemia development.


The patterns and dynamics of genomic instability in metastatic pancreatic cancer.
Pancreatic cancer is an aggressive malignancy with a five-year mortality of 97-98%, usually due to widespread metastatic disease. Previous studies indicate that this disease has a complex genomic landscape, with frequent copy number changes and point mutations, but genomic rearrangements have not been characterized in detail. Despite the clinical importance of metastasis, there remain fundamental questions about the clonal structures of metastatic tumours, including phylogenetic relationships among metastases, the scale of ongoing parallel evolution in metastatic and primary sites, and how the tumour disseminates. Here we harness advances in DNA sequencing to annotate genomic rearrangements in 13 patients with pancreatic cancer and explore clonal relationships among metastases. We find that pancreatic cancer acquires rearrangements indicative of telomere dysfunction and abnormal cell-cycle control, namely dysregulated G1-to-S-phase transition with intact G2-M checkpoint. These initiate amplification of cancer genes and occur predominantly in early cancer development rather than the later stages of the disease. Genomic instability frequently persists after cancer dissemination, resulting in ongoing, parallel and even convergent evolution among different metastases. We find evidence that there is genetic heterogeneity among metastasis-initiating cells, that seeding metastasis may require driver mutations beyond those required for primary tumours, and that phylogenetic trees across metastases show organ-specific branches. These data attest to the richness of genetic variation in cancer, brought about by the tandem forces of genomic instability and evolutionary selection.


Guidelines for using the QuantiFERON-TB Gold test for detecting Mycobacterium tuberculosis infection, United States.
On May 2, 2005, a new in vitro test, QuantiFERON-TB Gold (QFT-G, Cellestis Limited, Carnegie, Victoria, Australia), received final approval from the U.S. Food and Drug Administration as an aid for diagnosing Mycobacterium tuberculosis infection. This test detects the release of interferon-gamma (IFN-g) in fresh heparinized whole blood from sensitized persons when it is incubated with mixtures of synthetic peptides representing two proteins present in M. tuberculosis: early secretory antigenic target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10). These antigens impart greater specificity than is possible with tests using purified protein derivative as the tuberculosis (TB) antigen. In direct comparisons, the sensitivity of QFT-G was statistically similar to that of the tuberculin skin test (TST) for detecting infection in persons with untreated culture-confirmed tuberculosis (TB). The performance of QFT-G in certain populations targeted by TB control programs in the United States for finding latent TB infection is under study. Its ability to predict who eventually will have TB disease has not been determined, and years of observational study of substantial populations would be needed to acquire this information. In July 2005, CDC convened a meeting of consultants and researchers with expertise in the field to review scientific evidence and clinical experience with QFT-G. On the basis of this review and discussion, CDC recommends that QFT-G may be used in all circumstances in which the TST is currently used, including contact investigations, evaluation of recent immigrants, and sequential-testing surveillance programs for infection control (e.g., those for health-care workers). This report provides specific cautions for interpreting negative QFT-G results in persons from selected populations. This report is aimed at public health officials, health-care providers, and laboratory workers with responsibility for TB control activities in the United States.


Gating of sensory responses of descending brain neurones during walking in crickets
Single descending brain neurones were recorded and stained intracellularly in the neck connectives of crickets while they walked upon a styrofoam ball under open-loop conditions. The animal's translational and rotational velocities were measured simultaneously, and various stimuli were used to investigate the neuronal response characteristics. Stimulation with a moving grating or an artificial calling song of 5 kHz induced optomotor behaviour and positive phonotaxis. An acoustic stimulus of 20 kHz elicited negative phonotaxis. <P> We report the first clear evidence for behaviourally dependent gating of sensory responses of identified descending brain neurones. Most descending cells only responded to visual stimuli or to an artificial calling song of 5 kHz while the animal was walking, indicating that the responses to these stimuli were gated by the walking activity of the animal. In contrast to this, responses to stimuli that elicit negative phonotaxis, such as acoustic stimuli of 20 kHz, were not gated. This indicates that the gating of sensory responses in these cells depends on the behavioural context of the stimulus. <P> From these findings, we conclude that significant information about the properties of sensory processing in higher-order neurones can only be gained from tests in behaviourally relevant paradigms. Important characteristics might otherwise be missed, thus leading to misinterpretations regarding their function. <P>


Patterns in blood pressure medication use in US incident dialysis patients over the first 6 months.
Several observational studies have evaluated the effect of a single exposure window with blood pressure (BP) medications on outcomes in incident dialysis patients, but whether BP medication prescription patterns remain stable or a single exposure window design is adequate to evaluate effect on outcomes is unclear. We described patterns of BP medication prescription over 6 months after dialysis initiation in hemodialysis and peritoneal dialysis patients, stratified by cardiovascular comorbidity, diabetes, and other patient characteristics. The cohort included 13,072 adult patients (12,159 hemodialysis, 913 peritoneal dialysis) who initiated dialysis in Dialysis Clinic, Inc., facilities January 1, 2003-June 30, 2008, and remained on the original modality for at least 6 months. We evaluated monthly patterns in BP medication prescription over 6 months and at 12 and 24 months after initiation. Prescription patterns varied by dialysis modality over the first 6 months; substantial proportions of patients with prescriptions for beta-blockers, renin angiotensin system agents, and dihydropyridine calcium channel blockers in month 6 no longer had prescriptions for these medications by month 24. Prescription of specific medication classes varied by comorbidity, race/ethnicity, and age, but little by sex. The mean number of medications was 2.5 at month 6 in hemodialysis and peritoneal dialysis cohorts. This study evaluates BP medication patterns in both hemodialysis and peritoneal dialysis patients over the first 6 months of dialysis. Our findings highlight the challenges of assessing comparative effectiveness of a single BP medication class in dialysis patients. Longitudinal designs should be used to account for changes in BP medication management over time, and designs that incorporate common combinations should be considered.


Modulation transfer of the human eye as a function of retinal eccentricity.
We measured the monochromatic image quality of the eye across a wide visual field (120 degrees), with natural pupil (4 mm) and accommodation (3 diopters). The method is based on the acquisition and the posterior processing of double-pass aerial images of a point source imaged on the retina, which was kept at a fixed distance from the eye at all retinal eccentricities. The two-dimensional modulation transfer functions (MTF's) computed from the aerial images show that astigmatism is the dominant monochromatic aberration in both the fovea and the periphery and is also the major cause of variability among individuals. We found a slower decline in optical quality with eccentricity than had been found by previous measurements. Our foveal results are in close agreement with those of Campbell and Gubisch [J. Physiol. (London) 186, 558-578 (1966)], but off-axis optical quality is much better than found previously by Jennings and Charman [Am. J. Optom. Physiol. Opt. 55, 582-590 (1978); Vision Res. 21, 445-454 (1981)]. The optical system of the eye seems to follow a wide-angle lens design: the optical quality in the center (fovea) is not particularly good (it is far from the diffraction limit at this pupil size), but the modulation transfer function remains roughly constant for a wide visual field.


Monitoring bacterial community structure and variability in time scale in full-scale anaerobic digesters.
Using a high-throughput pyrosequencing technology, this study assessed bacterial community structure and time-scale variability in great detail in seven full-scale anaerobic digesters operated variously in terms of influent substrate, digestion temperature, and reactor configuration. Pyrosequencing generated a total of 83,774 sequence reads from 40 digester sludge samples collected monthly for six months. The highest number of sequence reads were detected within Proteobacteria (20.5%), followed by those within Bacteroidetes (19.7%), Firmicutes (17.8%), and Chloroflexi (4.8%). The relative composition of bacterial populations was varied within the digesters as well as between the digesters, and the bacterial community structures were mainly influenced by digestion temperature. Detailed bacterial community structures were assessed by analyzing the operational taxonomic units (OTUs) based on 97% sequence similarity, which resulted in a total of 9051 OTUs. Among these, a total of 31 core OTUs were analyzed and inferred phylogenetically, which enabled us to classify the sequences within an unclassified phylum. Unclassified sequences were mostly affiliated with the sequences within Spirochaetes and Firmicutes. Interestingly, numerically dominant novel phylotypes (18% of the total sequence reads) presumably involved in anaerobic digestion within Spirochaetes were identified. Temporal variability was further explored using a non-metric multidimensional scaling ordination which demonstrated that the variability of the bacterial community within the digesters was smaller than between digesters. Correlation analysis demonstrated that digester performance and operational conditions affected the pattern of bacterial community in the ordination. Additionally, a multi-response permutation procedure revealed that the bacterial communities within the digesters were more similar than those belonging to other digesters statistically, demonstrating a patchiness of the digesters in the distribution of bacterial populations. Overall, this study revealed the correlation of bacterial community structure and time-scale variability with digester performance and operating conditions.


Health-related quality of life in patients with idiopathic pulmonary fibrosis--cross-sectional and longitudinal study.
To validate the cross-sectional and longitudinal use of the Medical Outcome Study Short Form 36 (SF-36) for measuring health-related quality of life (HRQL) in patients with idiopathic pulmonary fibrosis (IPF). Patients were administered the SF-36 and concomitantly underwent laboratory and physiologic tests and high-resolution computed tomography (HRCT). Forty-six patients participated in the initial cross-sectional study, and 32 patients who were available more than one year later again underwent these studies under the same conditions. Patients with IPF had significantly lower scores across all 8 domains of the SF-36 when compared with the general population. Significant decline of HRQL was observed in 2 physical domains. There were significant differences in within-subject changes in a few domains according to worsening of the physiologic parameters. Vital capacity as percent of predicted was significantly correlated with the results of 6 subscales and its changes were significantly correlated with those of 4 subscales. The 6-min-walk distance was correlated significantly with 3 subscales and its changes were significantly correlated with those of 4 subscales. Changes in the HRCT ground-glass score were significantly correlated with those of 3 subscales. No significant correlations between changes in 3 domains and those of any clinical parameters were observed. Patients with IPF had significantly impaired HRQL in both physical and psychological functions. This disease clearly decreased the physical aspects of HRQL over time. HRQL instruments should be incorporated into routine evaluations of IPF patients, since they measure dimensions not fully estimated by clinical assessment.


Asymptomatic carotid endarterectomy. Patient and surgeon selection.
The applicability of prospective carotid endarterectomy protocols to the general population has been questioned. Outcomes for asymptomatic patients undergoing carotid endarterectomy were compared with the results of the Asymptomatic Carotid Atherosclerosis Study (ACAS) patients treated concurrently at our institution. Asymptomatic patients undergoing carotid endarterectomies (n = 277) from 1987 to 1993 (ACAS enrollment period) were reviewed. Primary end points were mortality, myocardial infarction, and stroke. Five subgroups were studied: (1) ACAS surgical patients; (2) ACAS-eligible patients not enrolled and ACAS surgeons; (3) ACAS-eligible patients not enrolled and non-ACAS surgeons; (4) ACAS-ineligible patients and ACAS surgeons; and (5) ACAS-ineligible patients and non-ACAS surgeons. ACAS-eligible patients were younger (P = .014), had more severe carotid stenosis (P = .001), and had lower incidences of pulmonary (P = .015) and renal (P = .008) diseases compared with ineligible patients. Patient selection (ACAS eligibility) significantly improved outcomes for mortality (P = .014) and myocardial infarction (P = .006). Length of stay favored ACAS-eligible patients (P = .004). ACAS surgeons operated on more severely stenotic carotid lesions (P = .005) and on patients with a lower incidence of coronary artery disease (P = .007). There was no difference in outcomes between ACAS and non-ACAS surgeons. Patient selection was a significant factor in determining outcome. With strict adherence to ACAS enrollment guidelines, the conclusions of ACAS appear applicable to patients seen at our institution with asymptomatic carotid stenosis.


Geometric correlations and breakdown of mesoscopic universality in spin transport.
We construct a unified semiclassical theory of charge and spin transport in chaotic ballistic and disordered diffusive mesoscopic systems with spin-orbit interaction. Neglecting dynamic effects of spin-orbit interaction, we reproduce the random matrix theory results that the spin conductance fluctuates universally around zero average. Incorporating these effects into the theory, we show that geometric correlations generate finite average spin conductances, but that they do not affect the charge conductance to leading order. The theory, which is confirmed by numerical transport calculations, allows us to investigate the entire range from the weak to the previously unexplored strong spin-orbit regime, where the spin rotation time is shorter than the momentum relaxation time.


Targeting human DNA polymerase alpha for the inhibition of keratinocyte proliferation. Part 1. Homology model, active site architecture and ligand binding.
In order to understand the binding modes of human DNA polymerase alpha (pol alpha) inhibitors on a molecular level, a 3D homology model of the active site of the enzyme was proposed based on the application of molecular modelling methods and molecular dynamic simulations using available crystal coordinates of pol alpha relatives. Docking results for a series of known nucleotide analogue inhibitors were consistent with reported experimental binding data and offered the possibility to elucidate structure-activity relationships via investigations of active site-inhibitor interactions. Furthermore, the study could explain, at least partially, the inhibitory effect of aphidicolin on pol alpha. In molecular dynamics simulations, aphidicolin occupied the catalytic centre, but acted in a not truly competitive manner with respect to nucleotides. It destabilized the replicating "closed" form of the pol alpha and transferred the enzyme into the inactive "open" conformation. This result is consistent with recent experiments on the binding mode of aphidicolin.


Effect of a Cognitive-Behavioral Prevention Program on Depression 6 Years After Implementation Among At-Risk Adolescents: A Randomized Clinical Trial.
Adolescents whose parents have a history of depression are at risk for developing depression and functional impairment. The long-term effects of prevention programs on adolescent depression and functioning are not known. To determine whether a cognitive-behavioral prevention (CBP) program reduced the incidence of depressive episodes, increased depression-free days, and improved developmental competence 6 years after implementation. A 4-site randomized clinical trial compared the effect of CBP plus usual care vs usual care, through follow-up 75 months after the intervention (88% retention), with recruitment from August 2003 through February 2006 at a health maintenance organization, university medical centers, and a community mental health center. A total of 316 participants were 13 to 17 years of age at enrollment and had at least 1 parent with current or prior depressive episodes. Participants could not be in a current depressive episode but had to have subsyndromal depressive symptoms or a prior depressive episode currently in remission. Analysis was conducted between August 2014 and June 2015. The CBP program consisted of 8 weekly 90-minute group sessions followed by 6 monthly continuation sessions. Usual care consisted of any family-initiated mental health treatment. The Depression Symptoms Rating scale was used to assess the primary outcome, new onsets of depressive episodes, and to calculate depression-free days. A modified Status Questionnaire assessed developmental competence (eg, academic or interpersonal) in young adulthood. Over the 75-month follow-up, youths assigned to CBP had a lower incidence of depression, adjusting for current parental depression at enrollment, site, and all interactions (hazard ratio, 0.71 [95% CI, 0.53-0.96]). The CBP program's overall significant effect was driven by a lower incidence of depressive episodes during the first 9 months after enrollment. The CBP program's benefit was seen in youths whose index parent was not depressed at enrollment, on depression incidence (hazard ratio, 0.54 [95% CI, 0.36-0.81]), depression-free days (d = 0.34, P = .01), and developmental competence (d = 0.36, P = .04); these effects on developmental competence were mediated via the CBP program's effect on depression-free days. The effect of CBP on new onsets of depression was strongest early and was maintained throughout the follow-up period; developmental competence was positively affected 6 years later. The effectiveness of CBP may be enhanced by additional booster sessions and concomitant treatment of parental depression. clinicaltrials.gov Identifier:NCT00073671.


A ruptured large aneurysm of the ductus arteriosus.
A 60 year old man was admitted with acute back pain followed by hoarseness. An aneurysm of the ductus arteriosus Botalli was diagnosed and an operation was recommended because of the high risk of complications such as rupture, embolism, or infection. The patient and his family refused surgery, however. The patient died suddenly of a rupture of the aneurysm a year later; necropsy confirmed the diagnosis. Several diagnostic methods were used and enhanced computed tomography gave the best representation of the aneurysm as it was seen at necropsy. This case indicates that enhanced computed tomography is probably the most useful investigation in patients with this type of aneurysm and it confirms the importance of an aneurysmectomy.


Kinematics of stair descent in young and older adults and the impact of exercise training.
Stair descent is a challenging task in old age. This study firstly investigated lower extremity kinematics during stair descent in young (YOU) and healthy, community dwelling older adults (OLD). Secondly, the impact of an exercise training intervention on age-related differences in stair descent was assessed. At baseline, a motion analysis system was used to determine spatio-temporal gait variables and lower extremity kinematics as YOU (n=23, age=27+/-3 years) and OLD (n=34, age=73+/-4 years) descended a three step staircase. The older adults were then divided into training (TRA) and control (CON) groups. For 12 months, TRA performed resistance, aerobic, balance, and flexibility exercises under supervision in a class environment (twice per week) and unsupervised at home (once per week). CON carried on with normal daily activities. Following the intervention, baseline measurements were repeated in TRA and CON. At baseline, total descent, stride cycle, and single support times were longer in OLD than in YOU. In addition, sagittal plane knee motion was lower in OLD whilst frontal and transverse plane pelvis and hip motion were higher in OLD. Exercise training did not reduce the age-related differences observed. In conclusion healthy older adults perform stair descent at a slower speed and with greater motion outside the plane of progression than young adults. We found no evidence that these differences are reduced by generic exercise training, at least in non-frail older adults.


Relationship between phase and energy methods for disparity computation.
The phase and energy methods for computing binocular disparity maps from stereograms are motivated differently, have different physiological relevances, and involve different computational steps. Nevertheless, we demonstrate that at the final stages where disparity values are made explicit, the simplest versions of the two methods are exactly equivalent. The equivalence also holds when the quadrature-pair construction in the energy method is replaced with a more physiologically plausible phase-averaging step. The equivalence fails, however, when the phase-difference receptive field model is replaced by the position-shift model. Additionally, intermediate results from the two methods are always quite distinct. In particular, the energy method generates a distributed disparity representation similar to that found in the visual cortex, while the phase method does not. Finally, more elaborate versions of the two methods are in general not equivalent. We also briefly compare these two methods with some other stereo models in the literature.


Nanomechanical properties of glucans and associated cell-surface adhesion of Streptococcus mutans probed by atomic force microscopy under in situ conditions.
This study used atomic force microscopy (AFM) to probe the local cell-surface interactions associated with the glucan polymers of Streptococcus mutans, the macromolecules most commonly attributed to the virulence of this microbe. In situ force spectroscopy was used to quantitatively probe and correlate cell-surface adhesion and dynamics with S. mutans UA140 wild-type and five glucosyltransferase mutants. Adhesion between the tooth surface and S. mutans is largely mediated by glucan production from sucrose via three glucosyltransferases (Gtfs; GtfB, GtfC and GtfD). To monitor the contribution of these particular Gtfs, isogenic mutants of S. mutans were constructed by specific gene inactivation and compared to the wild-type under sucrose and non-sucrose conditions. We report direct measurement of the mechanical properties associated with glucan macromolecules demonstrating that the local adhesion strength increases in a time-dependent process, with a decrease in the average number of rupture events. This finding suggests that S. mutans attaches mainly through glucans to surfaces in the presence of sucrose. In addition, a possible role of the Gtf proteins in sucrose-independent attachment is supported by the decreased adhesion properties of the GtfBCD mutant compared to the wild-type.


Intra-arterial thrombolysis for acute stroke in patients 80 and older: a comparison of results in patients younger than 80 years.
Intra-arterial fibrinolytic therapy is a promising treatment for acute ischemic stroke. Few data are available on its use in elderly patients. The purpose of this study was to compare the baseline characteristics, complications, and outcomes between intra-arterially treated ischemic stroke patients aged > or = 80 years and their younger counterparts. Patients aged > or = 80 years (n = 33) were compared retrospectively with contemporaneous patients aged < 80 years (n = 81) from a registry of consecutive patients treated with intra-arterial thrombolysis over a 9-year period. The very elderly and younger cohorts were very similar in baseline characteristics, including pretreatment stroke severity (National Institutes of Health Stroke Scale [NIHSS] 17 versus 16), differing only in history of stroke/transient ischemic attack (42% versus 22%, P = .01) and weight (66.8 versus 75.8 kg; P = .02). Significant differences in recanalization (TIMI 2-3) rates could not be detected between the very elderly and younger patients (79% versus 68%, P = .10). Rates of major symptomatic hemorrhage (7% versus 8%) and any intracerebral hemorrhage (39% versus 37%) did not differ. Outcomes at 90 days showed lower rates of excellent functional outcome (mRS < or = 1, 26% versus 40%, P = .02) and survival (57% versus 80%, P = .01) among the very elderly. Intra-arterial fibrinolysis in the elderly can be accomplished with recanalization rates and hemorrhage rates equal to that in younger patients. Although mortality rates are higher and good functional outcomes are lower than in younger persons, nondisabling outcomes may be achieved in a quarter of patients. These findings suggest that the investigation and use of intra-arterial thrombolytic treatment in very elderly patients should not be avoided but pursued judiciously.


Successful percutaneous transluminal angioplasty in native coarctation of aorta in young adult: a case report.
Coarctation of aorta is a rare cause of secondary hypertension, premature death will occur if no appropriate treatment is given. Before 1980, the only effective treatment was surgery, and recurrent stenosis was frequent. After then several works on percutaneous transluminal angioplasty of this disease were reported, with promising short-term and long-term results, especially in children and young adults. This report concerns a 19-year-old male who had hypertension for four years; the hypertension was caused by coarctation of the aorta, confirmed by cardiac catheterization and angiography. Average blood pressure (BP) of his upper extremities was 200/110 mmHg which dropped to 150/90 mmHg immediately, and still two years, after angioplasty, whereas BP of the lower extremities maintained at about 150/80 mmHg measured by Korotkoff technique. Follow-up transesophageal echocardiography found no coarctation or aneurysm at the dilatation site one year later. Magnetic resonance imaging, performed two years later, showed an almost normal aortic wall at the coaxctation. The postoperative course was uneventful, and patient follow-up at the cardiology clinic has regularly shown an average BP around 150/80 mmHg.


MR imaging of bone marrow edema and joint effusion in patients with osteonecrosis of the femoral head: relationship to pain.
Our aim was to determine the occurrence of bone marrow edema and joint effusion and their relationship to pain in patients with osteonecrosis of the femoral head on the basis of MR imaging. There were 71 patients with osteonecrosis of the femoral head based on characteristic radiographic and MR imaging findings. All patients had surgical confirmation of the disease. Both hips were affected with osteonecrosis in 39 patients, whereas only one hip was involved in 31 patients. The last patient underwent an arthroplasty of one hip during the study and had only one hip imaged. We evaluated a total of 110 hips in this study, of which 98 were painful. We staged osteonecrosis of the femoral head, using the classification of Steinberg et al. The 31 unaffected hips served as controls. Bone marrow edema and joint fluid were evaluated on MR images. Bone marrow edema was defined as an ill-defined area of low signal intensity on T1-weighted images with corresponding high signal intensity on T2-weighted or inversion recovery images localizing to the femoral head, neck, and intertrochanteric region. The amount of joint fluid was graded from 0 to 3. The peak of bone marrow edema occurred in stage III disease (72%); its odds ratio was seven times greater than that for stage I osteonecrotic hips. Effusions of a grade greater than or equal to 2 were seen most often in stage III disease (92%), compared with 10% in the control hips. With an effusion, bone marrow edema was 12.6 times greater when the hip was painful than when it was not. Both bone marrow edema and joint effusions existed with a peak occurrence in stage III disease. Bone marrow edema seems to have a stronger association with pain than does joint effusion in osteonecrosis of the femoral head.


The CogBIAS longitudinal study of adolescence: cohort profile and stability and change in measures across three waves.
Adolescence is a time of considerable social, cognitive, and physiological development. It reflects a period of heightened risk for the onset of mental health problems, as well as heightened opportunity for flourishing and resilience. The CogBIAS Longitudinal Study (CogBIAS-L-S) aims to investigate psychological development during adolescence. We present the cohort profile of the sample (N = 504) across three waves of data collection, when participants were approximately 13, 14.5, and 16 years of age. Further, we present descriptive statistics for all of the psychological variables assessed including (a) the self-report mood measures, (b) the other self-report measures, and (c) the behavioural measures. Differential and normative stability were investigated for each variable, in order to assess (i) measurement reliability (internal consistency), (ii) the stability of individual differences (intra-class correlations), and (iii) whether any adolescent-typical developmental changes occurred (multilevel growth curve models). Measurement reliability was good for the self-report measures (> .70), but lower for the behavioural measures (between .00 and .78). Differential stability was substantial, as individual differences were largely maintained across waves. Although, stability was lower for the behavioural measures. Some adolescent-typical normative changes were observed, reflected by (i) worsening mood, (ii) increasing impulsivity, and (iii) improvements in executive functions. The stability of individual differences was substantial across most variables, supporting classical test theory. Some normative changes were observed that reflected adolescent-typical development. Although, normative changes were relatively small compared to the stability of individual differences. The development of stable psychological characteristics during this period highlights a potential intervention window in early adolescence.


Low-molecular-weight heparin inhibits hypoxic pulmonary hypertension and vascular remodeling in guinea pigs.
We have shown previously that antiproliferative unfractionated heparins block hypoxia-induced pulmonary arterial hypertension (PAH) and vascular remodeling, and hypothesized that low-molecular-weight heparins (LMWHs) would too. To determine the potential role and mechanisms of dalteparin and enoxaparin (two LMWHs) in inhibiting hypoxic PAH and vascular remodeling. Male Hartley guinea pigs were exposed for 10 days to normobaric 10% oxygen with dalteparin (5 mg/kg), enoxaparin (5 mg/kg), or with an equivalent volume of normal saline solution. Normoxic control animals (n = 5) received room air for 10 days. Bovine pulmonary artery smooth-muscle cells (PASMCs) were grown in 10% fetal bovine serum without heparin, with dalteparin (1 microg/mL) or with enoxaparin (1 microg/mL). Pulmonary arterial pressure (PAP), cardiac index, right ventricular heart weight divided by left ventricular plus septum weight (RV/LV+S), hematocrit, percentage of wall thickness of intraacinar vessels (%WT-IA), percentage of wall thickness of terminal bronchiole vessels (%WT-TA), and the percentage of thick-walled vessels (%Thick) were determined. In PASMCs, expression of p27 and cell growth were compared because in mice whole heparin depends on p27 for its antiproliferative action. In hypoxic animals, hematocrit, PAP, total pulmonary vascular resistance index, RV/LV+S, %WT-IA, %WT-TA, and %Thick all rose significantly vs normoxic control animals (p < 0.05); cardiac index was unchanged. Dalteparin but not enoxaparin significantly reduced PAP, total pulmonary vascular resistance index, and RV/LV + S (p < 0.05 vs hypoxia alone); inhibited PASMC growth; and upregulated p27 expression. Enoxaparin moderately reduced vascular remodeling, which did not translate into less pulmonary hypertension. Not all LMWHs are the same. Dalteparin was more effective than enoxaparin in inhibiting pulmonary hypertension and vascular remodeling in hypoxic guinea pigs.


Anticancer effects of garlic and garlic-derived compounds for breast cancer control.
Garlic and garlic-derived compounds reduce the development of mammary cancer in animals and suppress the growth of human breast cancer cells in culture. Oil-soluble compounds derived from garlic, such as diallyl disulfide (DADS), are more effective than water-soluble compounds in suppressing breast cancer. Mechanisms of action include the activation of metabolizing enzymes that detoxify carcinogens, the suppression of DNA adduct formation, the inhibition of the production of reactive oxygen species, the regulation of cell-cycle arrest and the induction of apoptosis. Selenium-enriched garlic or organoselenium compounds provide more potent protection against mammary carcinogenesis in rats and greater inhibition of breast cancer cells in culture than natural garlic or the respective organosulfur analogues. DADS synergizes the effect of eicosapentaenoic acid, a breast cancer suppressor, and antagonizes the effect of linoleic acid, a breast cancer enhancer. Moreover, garlic extract reduces the side effects caused by anti-cancer agents. Thus, garlic and garlic-derived compounds are promising candidates for breast cancer control.


Shear bond strength of orthodontic brackets bonded using halogen light and light-emitting diode at different debond times.
The aim of this in vitro study was to compare the photoactivation effects of QTH (Quartz-Tungsten-Halogen) and LED (Light-Emitting Diode) on the SBS (Shear Bond Strength) of orthodontic brackets at different debond times. Seventy-two bovine lower incisors were randomly divided into two groups according to the photoactivation system used (QTH or LED). The enamel surfaces were conditioned with Transbond self-etching primer, and APC (Adhesive Pre-Coated) brackets were used in all specimens. Group I was cured with QTH for 20 s and Group II with LED for 10 s. Both groups were subdivided according to the different experimental times after bonding (immediately, 24 h and 7 days). The specimens were tested for SBS and the enamel surfaces were analyzed according to the Adhesive Remnant Index (ARI). The statistical analysis included the Tukey's test to evaluate the main effects of photoactivation and debond time on SBS. The Chi-square test was used to compare the ARI values found for each group, and no statistically significant difference was observed. The debond time of 7 days for QTH photoactivation showed statistically greater values of SBS when compared to the immediate and 24 h periods. There was no statistically significant difference between the QTH and LED groups immediately and after the 24 h period. In conclusion, bonding orthodontic brackets with LED photoactivation for 10 s is suggested because it requires a reduced clinical chair time.


Status of alcohol absorption in drinking drivers killed in traffic accidents.
One issue which constantly confronts the forensic toxicologist in drinking driver cases is the relationship between the breath or blood alcohol concentration (AC) of the driver at the time of an event such as a traffic stop or an accident and the AC measured at a time subsequent to the event. In theory, the AC can be rising, on a plateau or declining at the time of the event. Several studies have indicated that the overwhelming majority of drinking drivers are on a plateau or are post-absorptive at the time of the event. In this study, driver fatality cases investigated by the Office of the Chief Medical Examiner, State of Maryland during a three-year period were reviewed. Included in this study were cases positive for alcohol in the blood at a cutoff of 0.01 g/dL and death occurring within 15 min of the accident. In fact, many of these deaths were instantaneous or near instantaneous based on the injuries documented by the medical examiner at autopsy. The blood and urine were analyzed for alcohol by head-space gas chromatography and urine AC to blood AC ratios were calculated. A total of 129 cases were included in this study. Eleven of the 129 cases (8.5%) had urine to blood AC ratio less than 1.0. It is likely that these individuals were in the absorptive phase at the time that the accident occurred. Thirty-two cases had a urine to blood AC ratio between 1.0 and 1.2 inclusive. In these cases, the subject could be viewed as in the plateau phase of the blood AC versus time curve. The remaining 86 cases had a urine to blood AC ratio greater than 1.2. This suggests that these individuals were in the post-absorptive state at the time of the accident. The information acquired from this study provides additional evidence to support the notion that the vast majority of individuals are not in the absorptive phase at the time of a traffic stop or an accident.


A Genome-Wide Functional Genomics Approach Identifies Susceptibility Pathways to Fungal Bloodstream Infection in Humans.
Candidemia, one of the most common causes of fungal bloodstream infection, leads to mortality rates up to 40% in affected patients. Understanding genetic mechanisms for differential susceptibility to candidemia may aid in designing host-directed therapies. We performed the first genome-wide association study on candidemia, and we integrated these data with variants that affect cytokines in different cellular systems stimulated with Candida albicans. We observed strong association between candidemia and a variant, rs8028958, that significantly affects the expression levels of PLA2G4B in blood. We found that up to 35% of the susceptibility loci affect in vitro cytokine production in response to Candida. Furthermore, potential causal genes located within these loci are enriched for lipid and arachidonic acid metabolism. Using an independent cohort, we also showed that the numbers of risk alleles at these loci are negatively correlated with reactive oxygen species and interleukin-6 levels in response to Candida. Finally, there was a significant correlation between susceptibility and allelic scores based on 16 independent candidemia-associated single-nucleotide polymorphisms that affect monocyte-derived cytokines, but not with T cell-derived cytokines. Our results prioritize the disturbed lipid homeostasis and oxidative stress as potential mechanisms that affect monocyte-derived cytokines to influence susceptibility to candidemia.


Performance analysis of professional, semiprofessional, and junior elite rugby league match-play using global positioning systems.
The aim of the present study was to examine the positional differences in physical performance measures of professional, semiprofessional, and junior elite rugby league match-play using portable Global Positioning Systems (GPSs). Twelve professional, 12 semiprofessional, and 18 junior elite male rugby league players were monitored during 5 regular-season competition matches using portable GPS software. The mean total distance traveled during professional (8,371 ± 897 m) and semiprofessional (7,277 ± 734 m) match-play was significantly (p < 0.05) greater than that traveled during elite junior (4,646 ± 978 m) match-play. Position-specific total distance traveled and distance traveled per minute of playing time were significantly (p < 0.05) less for junior elite backs (5,768 ± 765 m; 74 ± 11 m·min) and forwards (4,774 ± 564 m; 82 ± 5 m·min) in comparison to those in professional (backs: 8,158 ± 673 m; 101 ± 8 m·min and forwards: 8,442 ± 812 m; 98 ± 12 m·min) and semiprofessional (backs: 7,505 ± 765 m; 94 ± 8 m·min and forwards: 6,701 ± 678 m; 89 ± 8 m·min) match-play. Maximum running speed, maximum sprints, and total sprint distance traveled by professional players were all significantly (p < 0.05) greater than those traveled by junior elite players but not semiprofessional players during match-play. Professional backs and forwards performed significantly (p < 0.05) more maximum sprints and traveled greater total distance during match-play in comparison to semiprofessional and junior elite players. The present findings demonstrate minimal differences in the physical performance measures of professional and semiprofessional rugby league match-play. The position-specific performance characteristics of junior elite match-play indicate that current junior elite player-development pathways may not provide adequate preparation for players transitioning into professional competition.


Cysteine-Specific Labeling of Proteins with a Nitroxide Biradical for Dynamic Nuclear Polarization NMR.
Dynamic nuclear polarization (DNP) enhances the signal in solid-state NMR of proteins by transferring polarization from electronic spins to the nuclear spins of interest. Typically, both the protein and an exogenous source of electronic spins, such as a biradical, are either codissolved or suspended and then frozen in a glycerol/water glassy matrix to achieve a homogeneous distribution. While the use of such a matrix protects the protein upon freezing, it also reduces the available sample volume (by ca. a factor of 4 in our experiments) and causes proportional NMR signal loss. Here we demonstrate an alternative approach that does not rely on dispersing the DNP agent in a glassy matrix. We synthesize a new biradical, ToSMTSL, which is based on the known DNP agent TOTAPOL, but also contains a thiol-specific methanethiosulfonate group to allow for incorporating this biradical into a protein in a site-directed manner. ToSMTSL was characterized by EPR and tested for DNP of a heptahelical transmembrane protein, Anabaena sensory rhodopsin (ASR), by covalent modification of solvent-exposed cysteine residues in two (15)N-labeled ASR mutants. DNP enhancements were measured at 400 MHz/263 GHz NMR/EPR frequencies for a series of samples prepared in deuterated and protonated buffers and with varied biradical/protein ratios. While the maximum DNP enhancement of 15 obtained in these samples is comparable to that observed for an ASR sample cosuspended with ~17 mM TOTAPOL in a glycerol-d8/D2O/H2O matrix, the achievable sensitivity would be 4-fold greater due to the gain in the filling factor. We anticipate that the DNP enhancements could be further improved by optimizing the biradical structure. The use of covalently attached biradicals would broaden the applicability of DNP NMR to structural studies of proteins.


Helminth infections drive heterogeneity in human type 2 and regulatory cells.
Helminth infections induce strong type 2 and regulatory responses, but the degree of heterogeneity of such cells is not well characterized. Using mass cytometry, we profiled these cells in Europeans and Indonesians not exposed to helminths and in Indonesians residing in rural areas infected with soil-transmitted helminths. To assign immune alteration to helminth infection, the profiling was performed before and 1 year after deworming. Very distinct signatures were found in Europeans and Indonesians, showing expanded frequencies of T helper 2 cells, particularly CD161+ cells and ILC2s in helminth-infected Indonesians, which was confirmed functionally through analysis of cytokine-producing cells. Besides ILC2s and CD4+ T cells, CD8+ T cells and γδ T cells in Indonesians produced type 2 cytokines. Regulatory T cells were also expanded in Indonesians, but only those expressing CTLA-4, and some coexpressed CD38, HLA-DR, ICOS, or CD161. CD11c+ B cells were found to be the main IL-10 producers among B cells in Indonesians, a subset that was almost absent in Europeans. A number of the distinct immune profiles were driven by helminths as the profiles reverted after clearance of helminth infections. Moreover, Indonesians with no helminth infections residing in an urban area showed immune profiles that resembled Europeans rather than rural Indonesians, which excludes a major role for ethnicity. Detailed insight into the human type 2 and regulatory networks could provide opportunities to target these cells for more precise interventions.


Acute puerperal uterine inversion: case report.
Acute puerperal uterine inversion is a rare but potentially fatal obstetric emergency in which the uterine fundus collapses into the uterine cavity. Maternal mortality is high unless the condition is recognised and quickly corrected. The duration of time elapsed from moment of diagnosis to that of correction, along with rapid resuscitation measures are of utmost importance in its prognosis. A recent case managed successfully is described followed by a short review of the literature.


Old disease, new targets. Part-II, haematological malignancies.
In last two decades newer therapies in cancer treatment have emerged and have opened new horizons. New term of targeted therapy has emerged and for certain malignancies the paradigm has really changed after the introduction of these agents. We have learnt and have seen the outcome of some diseases after the addition of these monoclonal antibodies (MoABs) and tyrosine kinase inhibitors (TKIs). Rituximab a MoAB against CD-20 has really paved its role in the treatment of B-cell lymphomas and become the sort of standard therapy. The TKIs are newer agents available in a pill form and have inhibited many pathways at cellular level which are necessary for cancer development. Imatinib has really changed the prognosis and outcome of chronic myeloid leukemia (CML) remarkably. For those patients who develope intolerance to imatinib or their disease became resistant to the imatinib the newer agents like dasatinib and nilotinib are second line options. The major edge of these newer agents is more potency with low side-effect profile. The major concern remains the cost.


Alpha neurofeedback training improves SSVEP-based BCI performance.
Steady-state visual evoked potential (SSVEP)-based brain-computer interfaces (BCIs) can provide relatively easy, reliable and high speed communication. However, the performance is still not satisfactory, especially in some users who are not able to generate strong enough SSVEP signals. This work aims to strengthen a user's SSVEP by alpha down-regulating neurofeedback training (NFT) and consequently improve the performance of the user in using SSVEP-based BCIs. An experiment with two steps was designed and conducted. The first step was to investigate the relationship between the resting alpha activity and the SSVEP-based BCI performance, in order to determine the training parameter for the NFT. Then in the second step, half of the subjects with 'low' performance (i.e. BCI classification accuracy <80%) were randomly assigned to a NFT group to perform a real-time NFT, and the rest half to a non-NFT control group for comparison. The first step revealed a significant negative correlation between the BCI performance and the individual alpha band (IAB) amplitudes in the eyes-open resting condition in a total of 33 subjects. In the second step, it was found that during the IAB down-regulating NFT, on average the subjects were able to successfully decrease their IAB amplitude over training sessions. More importantly, the NFT group showed an average increase of 16.5% in the SSVEP signal SNR (signal-to-noise ratio) and an average increase of 20.3% in the BCI classification accuracy, which was significant compared to the non-NFT control group. These findings indicate that the alpha down-regulating NFT can be used to improve the SSVEP signal quality and the subjects' performance in using SSVEP-based BCIs. It could be helpful to the SSVEP related studies and would contribute to more effective SSVEP-based BCI applications.


Percutaneous autologous bone-marrow grafting for nonunions. Influence of the number and concentration of progenitor cells.
Bone marrow aspirated from the iliac crest contains progenitor cells that can be used to obtain bone-healing of nonunions. However, there is little available information regarding the number and concentration of these cells that are necessary to obtain bone repair. The purpose of this study was to evaluate the number and concentration of progenitor cells that were transplanted for the treatment of nonunion, the callus volume obtained after the transplantation, and the clinical healing rate. Marrow was aspirated from both anterior iliac crests, concentrated on a cell separator, and then injected into sixty noninfected atrophic nonunions of the tibia. Each nonunion received a relatively constant volume of 20 cm(3) of concentrated bone marrow. The number of progenitor cells that was transplanted was estimated by counting the fibroblast colony-forming units. The volume of mineralized bone formation was determined by comparing preoperative computerized tomography scans with scans performed four months following the injection. The aspirates contained an average (and standard deviation) of 612 +/- 134 progenitors/cm(3) (range, 12 to 1224 progenitors/cm(3)) before concentration and an average of 2579 +/- 1121 progenitors/cm(3) (range, 60 to 6120 progenitors/cm(3)) after concentration. An average total of 51 x 10(3) fibroblast colony-forming units was injected into each nonunion. Bone union was obtained in fifty-three patients, and the bone marrow that had been injected into the nonunions of those patients contained >1500 progenitors/cm(3) and an average total of 54,962 +/- 17,431 progenitors. The concentration (634 +/- 187 progenitors/cm(3)) and the total number (19,324 +/- 6843) of progenitors injected into the nonunion sites of the seven patients in whom bone union was not obtained were both significantly lower (p = 0.001 and p < 0.01, respectively) than those in the patients who obtained bone union. The volume of the mineralized callus measured at four months on the computerized tomography scans of the patients who had union ranged from 0.8 to 5.3 cm(3) (mean, 3.1 cm(3)). There was a positive correlation between the volume of mineralized callus at four months and the number (p = 0.04) and concentration (p = 0.01) of fibroblast colony-forming units in the graft. There was a negative correlation between the time needed to obtain union and the concentration of fibroblast colony-forming units in the graft (p = 0.04). Percutaneous autologous bone-marrow grafting is an effective and safe method for the treatment of an atrophic tibial diaphyseal nonunion. However, its efficacy appears to be related to the number of progenitors in the graft, and the number of progenitors available in bone marrow aspirated from the iliac crest appears to be less than optimal in the absence of concentration.


Children unable to perform screening tests in vision in preschoolers study: proportion with ocular conditions and impact on measures of test accuracy.
To examine the relative prevalence of ocular conditions among children who are unable to perform preschool vision screening tests and the impact on measures of screening test performance. Trained nurse and lay screeners each administered a Lea Symbols visual acuity (VA) test (Good-Lite, Inc., Steamwood, IL), Stereo Smile II test (Stereo Optical, Inc., Chicago, IL), and Retinomax Autorefractor (Right Manufacturing, Virginia Beach, VA), and SureSight Vision Screener (Welch Allyn, Inc., Skaneateles Falls, NY) examinations to 1475 children who later received a comprehensive eye examination to identify amblyopia, strabismus, significant refractive error, and unexplained reduced VA. The outcomes of the examination for children for whom screeners were unable to obtain results (Unables) were compared to the outcomes of children who passed and children who failed each screening test. When estimating sensitivity, specificity, and positive and negative predictive values (PPV and NPV), Unables were classified as either screening failures or screening passers. Less than 2% of children were classified as Unables for each test. The percentage with an ocular condition was at least two times higher for Unables than for screening passers for six of the eight modes of screening (P < 0.05). Considering Unables as screening failures, rather than screening passers, increased the estimate of sensitivity by 1% to 3% (depending on test) and decreased the estimate of specificity by 0% to 2%; PPV decreased by 0% to 4% for most tests, whereas NPV increased by <1%. Preschool children who are unable to perform VIP screening tests are more likely to have vision disorders than are children who pass the tests. Because < or =2% of children were unable to do each test, referring these children for an eye examination had little impact on the PPV and NPV of the tests, as administered in VIP.


Endpoints in oncology clinical trials.
In oncology clinical trials, many different endpoints can be used as primary or secondary endpoints. Advances in cancer treatment have provided longer survival outcomes, particularly in certain types of cancer. Overall survival is accepted as the gold standard endpoint for demonstrating clinical benefit; however, it is associated with some disadvantages such as requirement of long-term follow-up, requirement of higher number of patients, and high cost. Thus, the question "what is the most appropriate endpoint in clinical trials?" comes to mind. The present review discusses the endpoints in oncology clinical trials.


Aerating recycled water on mushroom composting sites affects its chemical analysis and the characteristics of odor emissions.
Recycled water (RW) containing compost leachate can be a potent source of foul odor on mushroom composting sites. Samples of RW were repeatedly collected from storage tanks and pits of 14 mushroom composting sites in England and Ireland. Relationships between the effects of submerged aeration of the RW, the chemical and microbial characteristics of the RW, and the odors emitted were investigated. Recycled water samples were analyzed for pH, electrical conductivity (EC), redox potential, and dissolved oxygen concentration after 7 to 14 d cold storage. Freeze-dried material from the RW samples was chemically profiled by pyrolysis gas chromatography-mass spectrometry (GC-MS), and the content of odor precursor compounds was determined by the ninhydrin colorimetric method. The headspace air of containerized RW samples was analyzed by thermal desorption GC-MS and with gas detection tubes and assessed for odor intensity (OI) and concentration by panelists and serial dilution olfactometry. The predominant odorants in the headspace and freeze-dried residues of RW samples were volatile sulfur-containing compounds and carboxylic acids. The headspace OI, EC, dry matter content, and redox potential of RW were interrelated. The headspace OI and combined concentration of hydrogen sulfide (H(2)S) + dimethyl sulfide of RW were correlated (R(2) = 0.635; P < 0.001). Prediction of the OI of RW by measuring RW EC and the concentration of headspace sulfides using gas detector tubes enables rapid and low-cost monitoring of RW from mushroom composting sites. Submerged aeration of RW in storage tanks or pits reduced the RW headspace air OI and the combined H(2)S + dimethyl sulfide concentration by 88%.


Three new fusidic acid derivatives and their antibacterial activity.
Two steroid acids, cephalosporin P1 and isocephalosporin P1, were isolated from Hapsidospora irregularis FERM BP-2511. These compounds are structurally related to fusidic acid. Their NMR data were completely assigned on the basis of the 2D NMR spectra. Incubation of these two compounds with Microbacterium oxydans CGMCC 1788 in Luria-Bertani broth yielded the same set of three new 3-dehydrogenated products, 3-keto-isocephalosporin P1, 3-keto-cephalosporin P1 and 6-deacetyl-3-keto-cephalosporin P1. The final pH of the bacterial culture was 9.0. Incubation of 3-keto-isocephalosporin P1 or 3-keto-cephalosporin P1 in Tris-HCl buffer (pH 9.0) revealed that these two compounds can convert to each other by shifting the acetyl group between C-6 and C-7. The acetyl group at C-6 or C-7 can also be removed by hydrolysis to yield the minor product 6-deacetyl-3-keto-cephalosporin P1. These fusidic acid derivatives were tested for the antibacterial activity against the Gram-positive pathogen Staphylococcus aureus. 3-Keto-cephalosporin P1 showed the highest activity among the five compounds, with a minimal inhibition concentration (MIC) of 4 μg/mL, which is more potent than the substrate cephalosporin P1. Both cephalosporin P1 and 3-keto-cephalosporin P1 were active against methicillin-resistant S. aureus, with the same MIC of 8 μg/mL.


Food habit of the juvenile of the Japanese newt Cynops pyrrhogaster.
The previous study showed that the red coloration of the ventral skin of the Japanese newt Cynops pyrrhogaster was associated with the number of carotenoid vesicles and the content of carotenoid in the pigment cell of the skin. To elucidate the mechanism for the red coloration of the skin of the newt, we studied the food habit of the juvenile from the Japanese newt Cynops pyrrhogaster. Sixty-two juveniles were collected in Fukue Island in Nagasaki Prefecture from November 2000 to May 2002 and divided into 2 groups according to the snout-vent length (SVL). Over 400 prey animals were obtained from the juveniles by stomach flushing. In the larger group (SVL>30.0mm), Collembola (45.4%) and Acari (12.6%), which are very common species of soil animals, were the prey animals dominant in number. In the group with the smaller SVL (<29.9mm), Collembola (30.4%) and Acari (25.4%) were in number as well. We also studied the food habit of the Japanese clouded salamander, Hynobius nebulosus. In the salamander, Doratodesmidae (56.5%) and Amphipoda (13%) were the prey animals dominant in number. Our results, taken together, suggest that the Japanese juvenile C. pyrrhogaster does not change its food habit as it grows, and that it eats soil animals common in its habitat. Moreover, the food habit of juvenile C. pyrrhogaster differs from that of H. nebulosus, although the juveniles of both species live in the same area.


Brain catecholamines and catecholamine-synthesizing enzymes in renovascular hypertension in the rat.
1. Noradrenaline content of several rat brain stem and hypothalamic nuclei falls transiently at 72 h after initiation of renovascular hypertension (one-kidney Goldblatt model). 2. Tyrosine hydroxylase activity is significantly reduced in posterior, paraventricular and periventricular nuclei of hypothalamus at this time but returns to control value by 7 days. 3. Treatment with hydrallazine, 5 mg/kg intraperitoneally, twice daily or methaoxamine, 5 mg/kg, three times daily for 3 days respectively raises and lowers the noradrenaline content of brain nuclei, suggesting that short-term changes in noradrenaline may be secondary to afferent baroreceptor input. 4. At later times after the development of renovascular hypertension (7 and 28 days) activity of phenylethanolamine-N-methyl transferase is increased in the nucleus of the solitary tract and the locus coeruleus. 5. Brain catecholamines may participate both early in the development and later in the maintenance of renovascular hypertension.


Significant residual fluorinated alcohols present in various fluorinated materials.
Polyfluorinated telomer alcohols and sulfonamides are classes of compounds recently identified as precursor molecules to the perfluorinated acids detected in the environment. Despite the detection and quantification of these volatile compounds in the atmosphere, their sources remain unknown. Both classes of compounds are used in the synthesis of various fluorosurfactants and incorporated in polymeric materials used extensively in the carpet, textile, and paper industries. This study has identified the presence of residual unbound fluoro telomer alcohols (FTOHs) in varying chain lengths (C6-C14) in several commercially available and industrially applied polymeric and surfactant materials. NMeFOSE, a perfluoroalkyl sulfonamido alcohol, was also detected in a commercially available carpet protector product. A method was developed to remove these residual compounds from polymeric and surfactant materials by dispersion in water and stripping of the volatiles using a constant flow of air and trapping on XAD resin. Using gas chromatography mass spectrometry analysis, it was determined that the fluorinated materials examined consist of 0.04-3.8% residual alcohols on a fluoro alcohol to dry mass basis. These values indicate that residual alcohols, left unreacted and unbound from the manufacturing process of fluorinated polymers and surfactants, could be a significant source of the polyfluorinated telomer alcohols and sulfonamides released into the environment. This study suggests that elimination or reduction of these residual alcohols from all marketed fluorinated polymers and fluorosurfactants is key in reducing the prevalence of perfluorinated acids formed in the environment.