How repetitive are genomes?
Genome sequences vary strongly in their repetitiveness and the causes for this are still debated. Here we propose a novel measure of genome repetitiveness, the index of repetitiveness, Ir, which can be computed in time proportional to the length of the sequences analyzed. We apply it to 336 genomes from all three domains of life. The expected value of Ir is zero for random sequences of any G/C content and greater than zero for sequences with excess repeats. We find that the Ir of archaea is significantly smaller than that of eubacteria, which in turn is smaller than that of eukaryotes. Mouse chromosomes have a significantly higher Ir than human chromosomes and within each genome the Y chromosome is most repetitive. A sliding window analysis reveals that the human HOXA cluster and two surrounding genes are characterized by local minima in Ir. A program for calculating the Ir is freely available at http://adenine.biz.fh-weihenstephan.de/ir/. The general measure of DNA repetitiveness proposed in this paper can be efficiently computed on a genomic scale. This reveals a broad spectrum of repetitiveness among diverse genomes which agrees qualitatively with previous studies of repeat content. A sliding window analysis helps to analyze the intragenomic distribution of repeats.


Using Practice Testing, Public Speaking, and Source Monitoring to Examine the Influences of Learning Strategies and Stress on Episodic Memory.
Prior research demonstrated that learning information via retrieval practice, which entails studying and taking practice tests, resulted in less memory impairment under stress than learning information via repeated studying. The present experiment combined three experimental procedures to further examine the memory mechanisms underlying the efficacy of retrieval practice in the context of stress. A list-discrimination task was implemented, in which participants learned two distinct wordlists. This was combined with a retrieval-practice manipulation, as half of the participants engaged in practice testing and half engaged in conventional studying during learning. A week later, participants underwent stress induction, using the Trier Social Stress Test. Before and after stress induction, participants completed tests of item and source memory (i.e., list discrimination). The combination of these three procedures yielded informative results: retrieval practice, in the context of stress, improved item memory but not source memory relative to conventional studying. Limitations and future directions for the use of this methodology are discussed.


The relative contributions of insect and bird pollinators to outcrossing in an African Protea (Proteaceae).
A useful, but seldom applied, measure of the effectiveness of different pollinators is their contribution to the rate of outcrossing. This measure is particularly useful in facultatively autogamous plants for which seed set cannot be used as a direct measure of pollinator effectiveness. We used selective exclusion experiments to assess the importance of insects for outcrossing in Protea caffra, a facultatively autogamous shrub with scented flowers that are visited frequently by both birds and insects (mainly beetles). Pollen loads on stigmas, pollen tube growth, seed set, seed mass, germination, and early seedling survivorship were similar for vertebrate-excluded and open-pollinated inflorescences. Pollen-supplementation mostly did not increase seed set, revealing resource limitation. Mean multilocus outcrossing rates, estimated using eight polymorphic allozyme loci, were similar for progeny from inflorescences excluded from bird visitors (0.65) and for those visited by both birds and insects (0.59). Wright's fixation indices indicated that the adult population is near Hardy-Weinberg equilibrium but differed markedly for maternal plants (F(IS) = -0.187 ± 0.065) and their early stage progeny (F(IS) = 0.258 ± 0.002). Since seed from self and cross hand-pollinations were equally viable in terms of germination, this discrepancy in F(IS) could be explained by inbreeding depression that occurs between germination and reproductive maturity. Since outcrossing rates were not reduced when birds were excluded, we infer that insects are effective agents of cross pollination in P. caffra. This helps to explain the evolution of traits associated with insect pollination, such as fruity floral scent, in this species.


Mixed osmolytes: the degree to which one osmolyte affects the protein stabilizing ability of another.
Mixtures of organic osmolytes occur in cells of many organisms, raising the question of whether their actions on protein stability are independent or synergistic. To investigate this question it is desirable to develop a system that permits evaluation of the effect of one osmolyte on the efficacy of another to either force-fold or denature a protein. A means of evaluating the efficacy of an osmolyte is provided by its m-value, an experimental quantity that measures the ability of the osmolyte to force a protein to unfold or fold. An experimental system is presented that enables evaluations of the m-values of osmolytes in the presence and absence of a second osmolyte. The experimental system involves use of a marginally stable protein in 10 mM buffer (pH 7, 200 mM salt, and 34 degrees C) that is at the midpoint of its native to denatured transition. These conditions enable determination of m-values for protecting and denaturing osmolytes in the presence and absence of a second osmolyte, permitting assessment of the extent to which the two osmolytes affect each other's efficacy. The two osmolytes investigated in this work are the denaturing osmolyte, urea, and the protecting osmolyte, sarcosine. Results show unequivocally that neither osmolyte alters the efficacy of the other in forcing the protein to fold or unfold-the osmolytes act independently on the protein despite their combined concentrations being in the multi-molar range. These osmolytes avoid altering one another's efficacy at these high concentrations because the number of osmolyte interaction sites on the protein is large and the binding constants are quite small. Consequently, the site occupancies are low enough in number that the two osmolytes neither compete nor cooperate in interacting with the protein.


On Pathology Laboratory Recruitment and Retention.
The specific aims of the study are to analyze relationships between the personality traits of laboratory professionals and choice of profession and preferred work settings. Data from practicing laboratory professionals were collected via a web-based survey tool to gather information about personality types, choice of profession, and work setting preferences among medical laboratory professionals. Results of the survey showed that INFJ (Introversion, Intuition, Feeling, Judging) is the most common medical laboratory personality type across the various laboratory work settings and that there are no significant differences between the practitioners' personality type and the choice of profession within pathology. The study revealed laboratorians from higher-volume laboratories were 1.2 times more likely to prefer Judging than lower-volume laboratories, and younger medical laboratory practitioners were 2.3 times more likely to gravitate toward Perceiving than their older counterparts. The results of this study shed light on how employers can build on the personality preferences of the laboratory workforce to improve personal job satisfaction and laboratory productivity, quality, and work culture. The research implications are useful for laboratory recruitment and retention.


Treatment of waste incinerator air-pollution-control residues with FeSO4: laboratory investigation of design parameters.
The key design parameters of a new process for treatment of air-pollution-control (APC) residues (the Ferrox-process) were investigated in the laboratory. The optimisation involved two different APC-residues from actual incinerator plants. The design parameters considered were: amount of iron oxide supplied, the liquid-to-solid ratio of the process, the separation of solids and wastewater, the sequence of material mixing, the possibilities of reuse of water, the feasibility of using secondary (brackish) water, and simple means to improve the wastewater quality. The investigation showed that an optimum process configuration could be obtained yielding a stabilised solid product with low leaching of heavy metals and a dischargable wastewater with high contents of salts (in order to remove salts from the solid product) and low concentrations of heavy metals. The amount of iron added to the APC-residues must be optimised for each residue. The overall water use can be limited to a L/S-ratio of 3 l kg(-1) including water used for washing of the treated products.


Rapid homogeneous immunoassay based on time-resolved Förster resonance energy transfer for serodiagnosis of acute hantavirus infection.
We recently introduced a homogeneous immunoassay based on time-resolved Förster resonance energy transfer (TR-FRET) elicited by fluorophore-labeled antigen and fluorophore-labeled protein L, bound by an immunoglobulin. As the first clinical application, we employ this approach (LFRET) in serodiagnosis of Puumala hantavirus (PUUV) infection. A reference panel containing serum from individuals with acute (n = 21) or past (n = 17) PUUV infection and from PUUV-seronegative individuals (n = 20) was used to define the parameters. The clinical assay performance was evaluated with a prospectively collected serum panel (panel 2; n = 153). Based on the results for panel 1, the threshold for positivity was set at a signal level that was 3-fold over background, while those with a signal <3-fold over the background level were considered PUUV seronegative. With panel 1, 20/21 acute- and 7/10 past-infection samples induced positive signals, compared to 0/20 seronegatives. With panel 2, a positive signal was obtained in 39/40 acute- and 4/10 past-infection samples, as opposed to 7/103 seronegatives. However, after IgG depletion, 58/61 acute-infection samples were LFRET positive, while all past-infection and seronegative samples were negative, corresponding to 100% specificity and 95% sensitivity in detection of acute PUUV infection. We demonstrate that the novel immunoassay is a promising tool for rapid serodiagnosis of acute Puumala virus infection.


Reliability, feasibility, and validity of the quality of interactions schedule (QuIS) in acute hospital care: an observational study.
Research into relational care in hospitals will be facilitated by a focus on staff-patient interactions. The Quality of Interactions Schedule (QuIS) uses independent observers to measure the number of staff-patient interactions within a healthcare context, and to rate these interactions as 'positive social'; 'positive care'; 'neutral'; 'negative protective'; or 'negative restrictive'. QuIS was developed as a research instrument in long term care settings and has since been used for quality improvement in acute care. Prior to this study, its use had not been standardised, and reliability and validity in acute care had not been established. In 2014 and 2015 a three - phase study was undertaken to develop and test protocols for the use of QuIS across three acute wards within one NHS trust in England. The phases were: (1) A pilot of 16 h observation which developed implementation strategies for QuIS in this context; (2) training two observers and undertaking 16 h of paired observation to inform the development of training protocols; (3) training four nurses and two lay volunteers according to a finalised protocol followed by 36 h of paired observations to test inter-rater agreement. Additionally, patients were asked to rate interactions and to complete a shortened version of the Patient Evaluation of Emotional Care during Hospitalisation (PEECH) questionnaire. Protocols were developed for the use of QuIS in acute care. Patients experienced an average of 6.7 interactions/patient/h (n = 447 interactions). There was close agreement between observers in relation to the number of interactions observed (Intraclass correlation coefficient (ICC) = 0.97) and moderate to substantial agreement on the quality of interactions (absolute agreement 73%, kappa 0.53 to 0.62 depending on weighting scheme). There was 79% agreement (weighted kappa 0.40: P < 0.001; indicating fair agreement) between patients and observers over whether interactions were positive, negative or neutral. Observers using clear QuIS protocols can achieve levels of agreement that are acceptable for the use of QuIS as a research instrument. There is fair agreement between observers and patients' rating of interactions. Further research is needed to explore the relationship between QuIS measures and reported patient experience.


[Thyroid dysfunction and amiodarone].
Although most patients remain clinically euthyroid, some develop amiodarone-induced hyperthyroidism (HPEAI) or hypothyroidism (HPOAI). The authors present a retrospective analysis of ten patients with amiodarone-induced thyroid dysfunction. Six patients were female and mean amiodarone intake was 17.7 months. HPOIA was more common (six patients). From all the patients with HPEAI, two had type 2, one had type 1, and one had type 3 hyperthyroidism. Symptoms suggestive of thyroid dysfunction occurred in five patients, most of them with HPOAI. In HPEAI, the most frequent symptom was exacerbation of arrhythmia (three patients). Discontinuation of amiodarone and treatment with levothyroxine was chosen in 83.3% of the HPOAI cases, while thyonamide treatment with corticosteroids and without amiodarone was the option in 75% of the HPEAI cases. There were three deaths, all in patients with HPEAI. HPEAI is potentially fatal. The clinical picture may be vague, so the thyroid monitoring is mandatory.


Specificity in Legume-Rhizobia Symbioses.
Most species in the Leguminosae (legume family) can fix atmospheric nitrogen (N₂) via symbiotic bacteria (rhizobia) in root nodules. Here, the literature on legume-rhizobia symbioses in field soils was reviewed and genotypically characterised rhizobia related to the taxonomy of the legumes from which they were isolated. The Leguminosae was divided into three sub-families, the Caesalpinioideae, Mimosoideae and Papilionoideae. Bradyrhizobium spp. were the exclusive rhizobial symbionts of species in the Caesalpinioideae, but data are limited. Generally, a range of rhizobia genera nodulated legume species across the two Mimosoideae tribes Ingeae and Mimoseae, but Mimosa spp. show specificity towards Burkholderia in central and southern Brazil, Rhizobium/Ensifer in central Mexico and Cupriavidus in southern Uruguay. These specific symbioses are likely to be at least in part related to the relative occurrence of the potential symbionts in soils of the different regions. Generally, Papilionoideae species were promiscuous in relation to rhizobial symbionts, but specificity for rhizobial genus appears to hold at the tribe level for the Fabeae (Rhizobium), the genus level for Cytisus (Bradyrhizobium), Lupinus (Bradyrhizobium) and the New Zealand native Sophora spp. (Mesorhizobium) and species level for Cicer arietinum (Mesorhizobium), Listia bainesii (Methylobacterium) and Listia angolensis (Microvirga). Specificity for rhizobial species/symbiovar appears to hold for Galega officinalis (Neorhizobium galegeae sv. officinalis), Galega orientalis (Neorhizobium galegeae sv. orientalis), Hedysarum coronarium (Rhizobium sullae), Medicago laciniata (Ensifer meliloti sv. medicaginis), Medicago rigiduloides (Ensifer meliloti sv. rigiduloides) and Trifolium ambiguum (Rhizobium leguminosarum sv. trifolii). Lateral gene transfer of specific symbiosis genes within rhizobial genera is an important mechanism allowing legumes to form symbioses with rhizobia adapted to particular soils. Strain-specific legume rhizobia symbioses can develop in particular habitats.


Cowpox virus encodes a fifth member of the tumor necrosis factor receptor family: a soluble, secreted CD30 homologue.
Cowpox virus (Brighton Red strain) possesses one of the largest genomes in the Orthopoxvirus genus. Sequence analysis of a region of the genome that is type-specific for cowpox virus identified a gene, vCD30, encoding a soluble, secreted protein that is the fifth member of the tumor necrosis factor receptor family known to be encoded by cowpox virus. The vCD30 protein contains 110 aa, including a 21-residue signal peptide, a potential O-linked glycosylation site, and a 58-aa sequence sharing 51-59% identity with highly conserved extracellular segments of both mouse and human CD30. A vCD30Fc fusion protein binds CD153 (CD30 ligand) specifically, and it completely inhibits CD153/CD30 interactions. Although the functions of CD30 are not well understood, the existence of vCD30 suggests that the cellular receptor plays a significant role in normal immune responses. Viral inhibition of CD30 also lends support to the potential therapeutic value of targeting CD30 in human inflammatory and autoimmune diseases.


Mortality of Eggs and Newly Hatched Larvae of Lobesia botrana (Lepidoptera: Tortricidae) Exposed to High Temperatures in the Laboratory.
The hypothesis that bunch-zone leaf removal reduces infestations of the European grapevine moth, Lobesia botrana (Denis & Schiffermüller) (Lepidoptera: Tortricidae), by increasing egg and larval mortality owing to sunlight exposure was evaluated in the laboratory by subjecting different egg stages (white, red-eyes, and black-head) and newly hatched larvae to high temperatures. Based on temperatures recorded in a northern Italian vineyard on sun-exposed berries belonging to south-west facing bunches, eggs were subjected to constant temperatures of 40 °C and 37 °C for one or two periods of 3 or 6 h, and to 24-h temperature cycle with peak of 40 °C. Larvae were exposed to 24-h high-temperature cycles with peaks of 35, 37, and 40 °C. The results showed partial egg mortality at 40 °C, increasing with exposure hours and periods, and as eggs matured. Egg mortality was not affected by exposure to 37 °C. Larval survival already decreased significantly at 37 °C and was even lower at 40 °C. These laboratory data are in agreement with the hypothesis that temperatures reached by berries exposed to sunlight cause egg and larval mortality. Data on egg and larval susceptibility to high temperatures have also implications for species distribution and effects of climate change.


Dipyreno- and diperyleno-anthracenes from glyoxylic Perkin reactions.
Twofold Perkin condensation of 2,5-dibromophenylene-1,4-diacetic acid with arylglyoxylic acids followed by cyclo-dehydrobromination leads to dipyreno- and diperyleno-anthracene tetraesters and diimides. The imides show surprisingly large absorption shifts versus the esters, illustrating that electron-withdrawing substituents at the anthracene unit efficiently impart long wavelength absorption in such electron-deficient graphene nanoribbon fragments.


First report of a strong association between genetic polymorphisms of the prion protein gene (PRNP) and susceptibility to chronic wasting disease in sika deer (Cervus nippon).
Prion diseases are incurable neurodegenerative disorders caused by proteinase K-resistant prion protein (PrPSc ) derived from normal prion protein (PrPC ) encoded by the prion protein gene (PRNP). Although the cervid PRNP gene plays a pivotal role in the pathological mechanism of chronic wasting disease (CWD), there is no existing association analysis between susceptibility to CWD and genetic polymorphisms of the PRNP gene in sika deer. We investigated genetic polymorphisms of the PRNP gene using amplicon sequencing in sika deer. In addition, to identify a genetic susceptibility factor, we compared the genotype, allele and haplotype frequencies of the PRNP gene between CWD-positive and CWD-negative sika deer. Furthermore, to assess the effect of the genetic polymorphisms on sika deer prion protein (PrP), we performed in silico analysis using PolyPhen-2, PROVEAN and AMYCO. Finally, we analysed the tertiary structure and electrostatic potential of sika deer PrP based on single nucleotide polymorphisms (SNPs) using the SWISS-MODEL and Swiss-PdbViewer programs. We found a total of 24 SNPs of the PRNP gene, including 22 novel SNPs (10 synonymous SNPs and 12 nonsynonymous SNPs), in sika deer. Among the nonsynonymous SNPs, we found a strong association of susceptibility to CWD with c.56G > A (Ser19Asn). In addition, we found that c.56G > A (Ser19Asn), c.296A > T (His99Leu) and c.560T > A (Val187Asp) were predicted to have damaging effects on sika deer PrP. Furthermore, we observed significant alterations in the electrostatic potential of sika deer PrP by genetic polymorphisms of the 187Asp allele. To the best of our knowledge, this was the first association study between genetic polymorphisms of the PRNP gene and susceptibility to CWD in sika deer.


Genomic aberrations in young and elderly breast cancer patients.
Age at breast cancer diagnosis is a known prognostic factor. Previously, several groups including ours have shown that young age at diagnosis is associated with higher prevalence of basal-like tumors and aggressive tumor phenotypes. Yet the impact of age at diagnosis on the genomic landscape of breast cancer remains unclear. In this study, we examined the pattern of somatic mutations, chromosomal copy number variations (CNVs) and transcriptomic profiles in young and elderly breast cancer patients. Analyses were performed on The Cancer Genome Atlas (TCGA) dataset. Patients with metastatic disease at diagnosis, classified as normal-like by PAM50 or had missing clinical information were excluded. Young patients were defined as ≤45 years of age, while elderly patients were those ≥70 years of age at breast cancer diagnosis. The remaining patients were classified as "intermediate". We evaluated the association between age at diagnosis and somatic mutations, CNV and gene expression in a logistic regression model adjusting for tumor size, nodal status, histology and breast cancer subtype. All analyses were corrected for multiple testing using the Benjamini-Hochberg approach. In this study, 125, 486 and 169 patients were ≤45, 46-69 and ≥70 years of age, respectively. Older patients had more somatic mutations (n = 44 versus 35 versus 31; P = 0.0009) and more CNVs, especially in ductal tumors (P = 0.02). Eleven mutations were independently associated with age at diagnosis, of which only GATA3 was associated with young age (15.2% versus 8.2% versus 9%; P = 0.003). Only two CNV events were independently associated with age, with more chr18p losses in older patients and more chr6q27 deletions in younger ones. Younger age at diagnosis was associated with higher expression of gene signatures related to proliferation, stem cell features and endocrine resistance. Age adds a layer of biological complexity beyond breast cancer molecular subtypes, classic pathological and clinical variables, worthy of further consideration in future drug development as we seek to refine therapeutic strategies in the era of personalized medicine.


Ion channels in human endothelial cells.
Ion channels were studied in human endothelial cells from umbilical cord by the patch clamp technique in the cell attached mode. Four different types of ion channels were recorded: i) potassium channel current that rectifies at positive potentials in symmetrical potassium solutions (inward rectifier); ii) low-conductance non-selective cation channel with a permeability ratio K:Na:Ca = 1:0.9:0.2; iii) high-conductance cation-selective channel that is about 100 times more permeable for calcium than for sodium or potassium; iv) high-conductance potassium channel with a permeability ratio K:Na = 1:0.05. The extrapolated reversal potential of the inwardly rectifying current was near to the potassium equilibrium potential. The slope conductance decreased from 27 pS in isotonic KCl solution to 7 pS with 5.4 mmol/l KCl and 140 mmol/l NaCl in the pipette but 140 mmol/l KCl in the bath. The low-conductance non-selective cation channel showed a single-channel conductance of 26 pS with 140 mmol/l Na outside, 28 pS with 140 mmol/l K outside, and rectified in inward direction in the presence of Ca (60 mmol/l Ca, 70 mmol/l Na, 2.7 mmol/l K in the pipette) at negative potentials. The current could be observed with either chloride or aspartate as anion. The high-conductance non-selective channel did not discriminate between Na and K. The single-channel conductance was about 50 pS. The extrapolated reversal potential was more positive than +40 mV (140 K or 140 Na with 5 Ca outside). Both the 26 and 50 pS channel showed a run-down, and they rapidly disappeared in excised patches. The high-conductance potassium channel with a single-channel conductance of 170 pS was observed only rarely. It reversed near the expected potassium equilibrium potential. The 26 pS channel could be stimulated with histamine and thrombin from outside in the cell-attached mode. Both the 26 pS as well as the 50 pS channel can mediate calcium flux into the endothelial cell.


Activation of the MAPK, ERK, following Leishmania amazonensis infection of macrophages.
IL-10 is a critical cytokine in determining host susceptibility to Leishmania spp. We previously demonstrated that macrophage-derived IL-10 could contribute to disease exacerbation, but the mechanisms whereby Leishmania infections led to IL-10 induction were not fully understood. In this study, we demonstrated that infection of macrophages with Leishmania amazonensis amastigotes led to the activation of the MAPK, ERK1/2. This activation was required, but not sufficient for IL-10 induction. In addition to ERK activation, an inflammatory stimulus, such as low m.w. hyaluronic acid from the extracellular matrix, must also be present. The combination of these two signals resulted in the superinduction of IL-10. We also demonstrated that IgG on the surface of Leishmania amastigotes was required to achieve maximal IL-10 production from infected macrophages. Surface IgG engages macrophage FcgammaR to induce ERK activation. Macrophages lacking FcgammaR, or macrophages treated with an inhibitor of spleen tyrosine kinase, the tyrosine kinase that signals via FcgammaR, failed to activate ERK and consequently failed to produce IL-10 following infection with Leishmania amastigotes. We confirmed that ERK1/2 activation led to the phosphorylation of histone H3 at the IL-10 promoter, and this phosphorylation allowed for the binding of the transcription factor, Sp1, to the IL-10 promoter. Finally, the administration of U0126, an inhibitor of ERK activation, to infected mice resulted in decreased lesion progression with reduced numbers of parasites in them. Thus, our findings reveal an important role of MAPK, ERK signaling in the pathogenesis of Leishmania infection.


[On prevention of post-surgery acute urinary retention].
The article highlights the issues of prevention of acute urinary retention after surgery. The authors present results of a study on the efficacy and safety of 1-blockers for the prevention of acute urinary retention after surgery. The results imply that the use of 1-blockers in pre- and postoperative period can significantly reduce the risk of postoperative ischuria, contribute to independent urination after a single urinary catheterization in patients who experience urinary retention, improve the postoperative period and promote recovery of patients.


Usefulness of NT-proBNP in the Follow-Up of Patients after Myocardial Infarction.
Since serial analyses of NT-proBNP in patients with acute coronary syndromes have shown that levels measured during a chronic, later phase are a better predictor of prognosis and indicator of left ventricular function than the levels measured during an acute phase, we sought to assess the association of NT-proBNP, measured 6 months after acute myocardial infarction (AMI), with traditional risk factors, characteristics of in-hospital and early postinfarction course, as well as its prognostic value and optimal cut-points in the ensuing 1-year follow-up. Fasting venous blood samples were drawn from 100 ambulatory patients and NT-proBNP concentrations in lithium-heparin plasma were determined using a one-step enzyme immunoassay based on the »sandwich« principle on a Dimension RxL clinical chemistry system (DADE Behring-Siemens). Patients were followed-up for the next 1 year, for the occurrence of new cardiac events. Median (IQR) level of NT-proBNP was 521 (335-1095) pg/mL. Highest values were mostly associated with cardiac events during the first 6 months after AMI. Negative association with reperfusion therapy for index infarction confirmed its long-term beneficial effect. In the next one-year follow-up of stable patients, multivariate Cox regression analysis revealed the independent prognostic value of NT-proBNP for new-onset heart failure prediction (p=0.014), as well as for new coronary events prediction (p=0.035). Calculation of the AUCs revealed the optimal NT-proBNP cut-points of 800 pg/mL and 516 pg/mL, respectively. NT-proBNP values 6 months after AMI are mainly associated with the characteristics of early infarction and postinfarction course and can predict new cardiac events in the next one-year follow-up.


A novel method of minimally invasive rectus abdominis muscle flap harvest: Laparoscopic surgeons take note.
The rectus abdominis muscle (RAM) is a workhorse flap to fill or repair abdominal defects. A drawback of an open RAM harvest is donor site morbidity, and minimally invasive techniques for flap harvesting have been previously proposed but involve vertical division of the rectus fascia. We present a case of a 52-year-old woman with a recurrent rectovaginal fistula in a radiated field treated with a laparoscopic low anterior resection with simultaneous RAM flap harvest utilising a single Pfannenstiel incision. Our novel modified laparoscopic-assisted RAM harvest technique prevents longitudinal violation of the anterior and posterior rectus sheaths, thereby promoting a quick recovery, improved cosmesis and decreased post-operative morbidity.


Higher-order assemblies of oligomeric cargo receptor complexes form the membrane scaffold of the Cvt vesicle.
Selective autophagy is the mechanism by which large cargos are specifically sequestered for degradation. The structural details of cargo and receptor assembly giving rise to autophagic vesicles remain to be elucidated. We utilize the yeast cytoplasm-to-vacuole targeting (Cvt) pathway, a prototype of selective autophagy, together with a multi-scale analysis approach to study the molecular structure of Cvt vesicles. We report the oligomeric nature of the major Cvt cargo Ape1 with a combined 2.8 Å X-ray and negative stain EM structure, as well as the secondary cargo Ams1 with a 6.3 Å cryo-EM structure. We show that the major dodecameric cargo prApe1 exhibits a tendency to form higher-order chain structures that are broken upon interaction with the receptor Atg19 in vitro The stoichiometry of these cargo-receptor complexes is key to maintaining the size of the Cvt aggregate in vivo Using correlative light and electron microscopy, we further visualize key stages of Cvt vesicle biogenesis. Our findings suggest that Atg19 interaction limits Ape1 aggregate size while serving as a vehicle for vacuolar delivery of tetrameric Ams1.


Historical biogeography of the highly diverse brown seaweed Lobophora (Dictyotales, Phaeophyceae).
The tropical to warm-temperate marine brown macroalgal genus Lobophora (Dictyotales, Phaeophyceae) recently drew attention because of its striking regional diversity. In this study we reassess Lobophora global species diversity, and species distributions, and explore how historical factors have shaped current diversity patterns. We applied a series of algorithmic species delineation techniques on a global mitochondrial cox3 dataset of 598 specimens, resulting in an estimation of 98-121 species. This diversity by far exceeds traditional diversity estimates based on morphological data. A multi-locus time-calibrated species phylogeny using a relaxed molecular clock, along with DNA-confirmed species distribution data was used to analyse ancestral area distributions, dispersal-vicariance-founder events, and temporal patterns of diversification under different biogeographical models. The origin of Lobophora was estimated in the Upper Cretaceous (-75 to -60 MY), followed by gradual diversification until present. While most speciation events were inferred within marine realms, founder events also played a non-negligible role in Lobophora diversification. The Central Indo-Pacific showed the highest species diversity as a result of higher speciation events in this region. Most Lobophora species have small ranges limited to marine realms. Lobophora probably originated in the Tethys Sea and dispersed repeatedly in the Atlantic (including the Gulf of Mexico) and Pacific Oceans. The formation of the major historical marine barriers (Terminal Tethyan event, Isthmus of Panama, Benguela upwelling) did not act as important vicariance events. Long-distance dispersal presumably represented an important mode of speciation over evolutionary time-scales. The limited geographical ranges of most Lobophora species, however, vouch for the rarity of such events.


Comparison of renal function indicators according to hydration volume in patients receiving intravenous acyclovir with CNS infection.
We aimed to compare the changes in renal function indicators as a function of hydration volume in patients treated with acyclovir for suspected herpes simplex virus (HSV) infection. We obtained data from 216 acyclovir-treated patients hospitalized between 2007 and 2012 for suspected HSV infection. Intravenous hydration volume and renal function indicators (serum creatinine [sCr], blood urea nitrogen [BUN], glutamate oxaloacetate transferase, glutamate pyruvate transferase, and uric acid levels; estimated glomerular filtration rate [eGFR]; and urine pH) were compared among the patients. The indicators were assessed before acyclovir infusion and after 3 days of acyclovir infusion. Before acyclovir infusion, all the indicators were within normal ranges in all groups (hydration volume lower than 2 L/day, higher than 2 L/day, and without hydration). After acyclovir infusion for 3 days, the groups without hydration and with a volume lower than 2 L/day showed increased sCr (2.22 ± 0.51 and 1.70 ± 0.35 mg/dl, respectively), BUN levels (28.33 ± 0.57 and 22.14 ± 7.95 mg/dl, respectively), and glutamate oxaloacetate transferase (48.00 ± 2.65 IU/L, without hydration) and eGFRs lower than the normal range (53.03 ± 3.05 and 59.66 ± 10.25 ml/min, respectively; p < .001 for all parameters). The renal function indicators were within normal limits in the group with a hydration volume higher than 2 L/day. Renal function indicators in acyclovir-treated patients varied according to hydration volume. Health care providers should consider whether the hydration volume in each patient receiving intravenous acyclovir is sufficient for preventing nephropathy.


Relief of jaundice by external beam radiotherapy and intraluminal brachytherapy in patients with extrahepatic cholangiocarcinoma: results without stenting.
To clarify whether external beam radiotherapy combined with intraluminal brachytherapy without stenting provides long-term relief of jaundice in extrahepatic cholangiocarcinoma patients. Twenty-five patients with unresectable hilar or distal cholangiocarcinoma were treated by external beam radiotherapy (30 or 50 Gy) combined with intraluminal brachytherapy (24 to 40 Gy). After radiotherapy, we removed the biliary drainage tubes from the patients who responded, and we did not perform stenting in these patients. In 19 (76%) patients, full patency was achieved at the treated lesion, and they were tube-free thereafter. The tube-free survival time in the 19 patients ranged from 7 to 468 days (median: 76 days). Cholangitis and gastroduodenal ulcer developed in 10 (40%) and 2 (8%) patients, respectively, as adverse events after the combined radiotherapy. The median survival time of all patients was 9.3 months, and their 6- and 12-month survival rates were 75.3% and 29.3%, respectively. The ratios of tube-free to overall survival in the 19 patients ranged from 2.4% to 79.4% (median: 26.8%). Eight patients died tube-free. Although there were limitations to the long-term relief of jaundice by this combined radiotherapy alone, tube-free status was achieved in some patients without stenting.


The Opioid Abuse Risk Screener predicts aberrant same-day urine drug tests and 1-year controlled substance database checks: A brief report.
The Opioid Abuse Risk Screener was developed to support well-informed decision-making in opioid analgesic prescribing by extending the breadth of psychiatric risk factors evaluated relative to other non-clinician-administered measures. We examined the preliminary predictive validity of the Opioid Abuse Risk Screener relative to the widely used Screener and Opioid Assessment for Patients with Pain-Revised in predicting aberrant urine drug tests and controlled substance database checks. The Opioid Abuse Risk Screener is significantly different from the Screener and Opioid Assessment for Patients with Pain-Revised in predicting aberrant same-day urine drug tests (Z = 2.912, p = 0.0036) and controlled substance database checks within 1 year of assessment (Z = 3.731, p = 0.0002). Promising preliminary analyses using machine learning methods are also discussed.


Relationship between heat shock protein induction and the binding of antibodies to the extractable nuclear antigens on cultured human keratinocytes.
The importance of environmental factors such as ultraviolet light and temperature in the pathogenesis of cutaneous lupus erythematosus is well recognized. Recent evidence suggests the presence of autoantibodies to heat shock proteins (HSP) in the sera and enhanced expression of the HSP70 gene in peripheral blood mononuclear cells of patients with systemic lupus erythematosus. We designed experiments to determine how HSP or stress protein inducers affect the cell surface binding of IgG antibodies from sera containing anti-SS-A/Ro and anti-ribonuclear protein (RNP) antibodies to keratinocytes because these antibodies are considered to be one of the immunologic triggers of cutaneous lupus erythematosus. Immunofluorescence and immunoblot analysis using a monoclonal antibody to the 72 kDa of HSP revealed that an 18-h incubation with 10 micrograms/ml of delta 12-PGJ2, one of cytotoxic prostaglandins, induced HSP72 formation in cultured human keratinocytes. delta 12-PGJ2 augmented the binding of IgG antibodies from sera containing anti-U1RNP and anti-SS-A/Ro antibodies to cultured keratinocytes, but produced no enhancement of the binding of IgG antibodies from sera containing anti-Sm or anti-DNA antibodies. Similar results were also obtained by using flow cytometry analysis. HSP was also induced by ultraviolet B irradiation. These results suggest that exposure of keratinocytes to stressors such as delta 12-PGJ2 and ultraviolet light increases the binding sites for U1RNP,SS-A/Ro, and SS-B/La antibodies. The association between HSP induction and the appearance of extractable nuclear antigens may provide a better understanding of why environmental stimuli can promote the development of erythematous lesions in the skin.


GC-MS profiling and bioactivity prediction of compounds from Momordica charantia L. extract.
Momordica species are vegetable crops, belonging to the family of Cucurbitaceae. For the present study plant samples of Momordica charantia were collected from different parts of Barak Valley of Assam, India. Hyphenated technique Gas Chromatography-Mass Spectroscopy (GC-MS) of the chromatographically separated components of the plant extract was performed to identify some of the principles. All the isolated compounds exhibited notable interaction with the enzyme Bone morphogenic protein upon in silico screening. All the compounds were screened in silico for toxicity and then docked with the target enzyme individually. It was found that these compounds are interacting with the target enzyme and is able to inhibit the enzyme activity more efficiently as compared to the reference ligand.


Functional upper airway obstruction.
Functional disorders of the vocal cords can present with acute, dramatic upper airway obstruction, with features mimicking asthma or functional dysphonia. We report the case of an eighteen-year-old female with acute airway obstruction initially misdiagnosed as asthma. Laryngoscopy, bronchoscopy and psychiatric assessment confirmed a diagnosis of functional disorder of the vocal cords. Although a diagnosis of exclusion, it must be borne in mind in order to reduce unnecessary medical therapy and surgical intervention. It is only with the correct diagnosis that patients can receive appropriate therapy.


[A fatal case of skin cancer 18 years after cadaveric donor renal transplantation].
A 39 years old man was admitted with chronic renal failure, and in June, 1968, he received renal transplantation from a cadaveric donor, 30 years old male, died from cerebral contusion. The graft had been well functioning for 18 years, and he had been engaged in his occupation. At the age of fifty-seven skin cancer (squamous cell carcinoma) occurred in his face, and surgical treatment was carried out. But it metastasized to left parotid, bilateral lungs and other areas of skin, and he died in October 1986, complicated by cerebral infarction. Some reports mention that the incidence of malignancies in posttransplant patients is greater than in general population. Our study reveals that immunological surveillance is weakened in posttransplant patients because of decrease of NK activity, but that the incidence of malignancies (male 0.78%, female 0.45%) is not so great as that reported. There is need of further investigation in this problem.


Retinoic acid induces differentiation of buffalo (Bubalus bubalis) embryonic stem cells into germ cells.
Development of precise and reproducible culture system for in vitro differentiation of embryonic stem (ES) cells into germ cells counts as a major leap forward for understanding not only the remarkable process of gametogenesis, otherwise obscured by limited availability of precursor primordial germ cells (PGCs), but in finally treating the catastrophic infertility. Taking into account the significant role of retinoic acid (RA) during in vivo gametogenesis, we designed the present study to investigate the effects of its stimulation on directing the differentiation of ES cells into germ cells. The effects of RA were analyzed across dose-and-time upon various stages of gametogenesis like PGC induction, meiosis initiation and completion, haploid cell formation and development of the final gamete (oocyte and spermatozoa). Out of the series of RA doses (2, 4, 8, 16, 20 and 30μM), 16μM RA for 8day culture interval was found to induce highest expression of PGC- and meiosis-associated genes like DAZL, VASA, SYCP3, MLH1, TNP1/2 and PRM2, while mature germ cell genes like BOULE and TEKT1 (Spermatocyte markers), GDF9 and ZP2 (Oocyte markers) showed higher expression at 2μM RA dose, suggesting functional concentration-gradient of RA activity. Immunocytochemistry revealed expression of germ lineage-specific markers like: c-KIT, DAZL and VASA (PGC-markers); SYCP3, MLH1 and PROTAMINE1 (Meiotic-markers); ACROSIN and HAPRIN (Spermatocyte-markers); and GDF9 and ZP4 (Oocyte-markers) in optimally differentiated embryoid bodies (EBs) and adherent cultures. We observed significantly reduced (p<0.05) concentration of 5-methyl-2-deoxycytidine in RA-differentiated EBs which is suggestive of the occurrence of methylation erasure. FACS analysis of optimally differentiated cultures detected 3.07% haploid cell population, indicating completion of meiosis. Oocyte-like structures (OLS) were obtained in adherent differentiated cultures. They had a big nucleus and a zona pellucida (ZP4) coat. They showed progression through 2-cell, 4-cell, 8-cell, morula and blastocyst-like structures upon extended culture beyond 14days.


1-Year Outcomes of a Multicenter Randomized Controlled Trial of the Ankura II Thoracic Endoprosthesis for the Endovascular Treatment of Stanford Type B Aortic Dissections.
The Ankura II Thoracic Stent Graft System (Lifetech, Shenzhen, China) is an evolution of the Ankura stent graft. This study reports one-year outcomes of the Ankura II Thoracic Stent Graft System for endovascular treatment of Stanford type B aortic dissections. The Ankura II Thoracic Aortic Endovascular Trial was a randomized, single-blinded, clinical trial conducted at 12 Chinese institutes. The enrolled patients diagnosed with Stanford type B aortic dissections (TBADs) were randomly assigned to the Ankura group or Ankura II group. Standard follow-up examinations were performed at 1, 6, and 12 months. Safety and efficacy data were analyzed. 132 patients with TBADs were enrolled. The outcomes for the primary safety end points revealed that the Ankura II stent graft was statistically non-inferior compared to the Ankura stent graft. The 1-month device-related major adverse events (1.6 vs. 0%; p = 0.48), 1-month all-cause mortality (1.7 vs. 4.5%; p = 0.621), 12-month survival rate (95.2 ± 2.7% vs. 94.1 ± 2.9%; p = 0.769), and major adverse event (MAE) rate (5.1 vs. 4.7% at 1 month; p = 0.73 and 5.8 vs. 8.9% at 12 months; p = 0.718) of Ankura II group are all comparable to Ankura group. The two groups showed similar primary effectiveness and true lumen expansion effect, and false lumen remodeling was improved in Ankura II group (-100.0 vs. -48.5%; p = 0.08). The one-year outcomes from this prospective, randomized, multicenter study demonstrate that Ankura II stent graft shows comparable results to Ankura for treating TBADs, resulting in low mortality rates, MAEs and reintervention rates. ChiCTR-TRC-12002844.


Differential levels of mRNAs for cytokines, the interleukin-2 receptor and class II DR/DQ genes in ovine interstitial pneumonia induced by maedi visna virus infection.
The relative levels of selected cytokine, interleukin-2 receptor, class II DR and DQ RNAs, and maedi visna virus (MVV) RNA were measured by reverse-transcriptase polymerase chain reaction (RT-PCR) in the lungs of sheep with natural maedi visna virus infection (n = 8) and a group of age/sex/breed-matched MVV seronegative sheep (n = 4). These animals were divided into two groups, irrespective of serostatus, according to the severity of lymphocytic interstitial pneumonia. The severity of lung lesions was determined by clinical sign, lung weight, and lesion sore in the lungs measured by three pathologic parameters. Sheep with lung lesions showed hyperelevated levels of granulocyte-macrophage colony-stimulating factor upregulated gamma-interferon, interleukin 2 receptor, and interleukins 1 beta, 4, and 10 mRNAs. Class II mRNAs were found not to be elevated in the lungs of sheep with lung lesions. Tumor necrosis factor alpha and transforming growth factor beta 1 mRNA levels were similar in all sheep lungs studied. We discuss the major roles played by granulocyte-macrophage colony-stimulating factor and type 2 cytokines in the pathogenesis of this disease and the possible stimulation of the production of these cytokines by viral surface glycoproteins.


Structure Elucidation and Identification of Novel Lead Molecules against Sulfur Import Protein cysA of Mycobacterium tuberculosis.
The present work considers the Sulphate import ABC transporter protein (cysA) as a potential drug target for the identification of inhibitors for the protein. The ABC (ATP binding cassette) transporters play a crucial role in the survival and virulence of Mycobacterium tuberculosis by the acquisition of micronutrients from host tissue. The 3D structural features of the cysA protein are built. Molecular scaffolds are identified by implementing active site identification, ADME properties, Virtual Screening, and a few other computational techniques. The theoretical model of cysA is predicted using homology modeling protocols, and the structure is validated by various validation methods. The prediction of partial dimer formation through protein-protein docking methods gave insight into the conformational changes taking place in the cysA protein. The natural substrate ATP is docked with cysA protein that confirms the ATP binding site. To find the drug-like compounds, virtual screening studies were carried out around the active site by several ligand databases. The findings demonstrate the significance of residues SER41, GLY42, ARG50, GLN85, HIS86, LYS91, ARG142, and ASP161 in drug-target interactions. The docking studies of existing TB drugs against cysA were also performed. The result analysis shows that none of the existing drugs inhibits the ATP active site, which confirms cysA as a promising drug target. Using in-silico methods, the ADME parameters of a few chosen ligand molecules are predicted and contrasted with the ADME characteristics of the available TB medications. The results revealed the values of ADME parameters of selected ligand molecules are more permissible than existing TB drugs, which emphasizes the drug-like activity of ligand molecules by inhibition of cysA proteins. The structural data, active site information, and selected ligand molecules help in the identification of new therapeutic scaffolds for Tuberculosis.


The effect of sodium bisulfite on the melting profiles of nucleic acids and on their respective nucleosides.
The effect of sodium bisulfite (0.27 M, pH 7) on melting behavior of DNA, yeast RNA and their respective nucleosides was studied. It was found that bisulfite added not only to pyrimidine bases but also to purine bases of nucleic acids and of nucleosides. The addition products were stable at higher temperatures but reverted to parent compounds at room temperature. The only exception was the addition product of uridine which was stable at room temperature and could be isolated by paper chromatography in a 42-62% yield. Heating of DNA solutions in the presence of bisulfite to 95 degrees C caused a 90% loss of absorbance at 260 nm. On cooling, the absorbance was essentially recovered. When compared to the melting behavior of DNA in 0.27 M NaCl or 0.09 M Na2SO4 (same ionic strength), it was found that bisulfite destabilized double helical structure of DNA and that reversible addition of bisulfite did occur much below the melting temperature of DNA observed in the other two solvents.


IgA rheumatoid factor as predictor of disease activity.
IgA rheumatoid factor (RF) was shown to occur in rheumatoid arthritis (RA) sera in 1963, but for almost 20 years thereafter this RF isotype received little attention. Solid-phase immunoassays, esp. ELISA, are now widely used for the detection of serum IgA RF which may be raised in RA, primary Sjögren's syndrome, systemic lupus erythematosus (SLE) and several other diseases. Over the past four years the following observations on IgA RF have been published: While IgM RF levels showed positive correlation with age, IgA RF levels did not. Gold therapy reduced IgA RF levels but the relationship to clinical improvement was not clear. In longitudinal studies, changes in overall disease activity, Ritchie articular index, grip strength and ESR showed a closer association with IgA RF than with IgM RF. Five groups have reported an association between IgA RF and the severity of erosive arthritis. In early RA, IgA RF may be a more specific predictor of disease severity than IgM RF or IgG RF. Further work is needed on the occurrence and biology of IgA RF, but measuring this isotype alone would improve the clinical specificity of routine RF tests.


Real-time gait event detection for lower limb amputees using a single wearable sensor.
This paper presents a rule-based real-time gait event/phase detection system (R-GEDS) using a shank mounted inertial measurement unit (IMU) for lower limb amputees during the level ground walking. Development of the algorithm is based on the shank angular velocity in the sagittal plane and linear acceleration signal in the shank longitudinal direction. System performance was evaluated with four control subjects (CS) and one transfemoral amputee (TFA) and the results were validated with four FlexiForce footswitches (FSW). The results showed a data latency for initial contact (IC) and toe off (TO) within a range of ± 40 ms for both CS and TFA. A delay of about 3.7 ± 62 ms for a foot-flat start (FFS) and an early detection of -9.4 ± 66 ms for heel-off (HO) was found for CS. Prosthetic side showed an early detection of -105 ± 95 ms for FFS whereas intact side showed a delay of 141 ±73 ms for HO. The difference in the kinematics of the TFA and CS is one of the potential reasons for high variations in the time difference. Overall, detection accuracy was 99.78% for all the events in both groups. Based on the validated results, the proposed system can be used to accurately detect the temporal gait events in real-time that leads to the detection of gait phase system and therefore, can be utilized in gait analysis applications and the control of lower limb prostheses.


Structural manifestations of proton transfer in complexes of 2,6-dichlorophenols with pyridines.
DFT B3LYP/6-31G(d,p) calculations were performed to describe the proton transfer reaction pathway in the 2,6-dichlorophenolate of pyridine. The aim of these calculations was to establish the character of the dependence of the structure parameters on the proton transfer and comparing the results with known structures, e.g. the 2,6-dichloro-4-nitro- and pentachlorophenolates of pyridines. To make this comparison more reliable, the calculations were repeated with the use of a reaction-field correction with the Onsager radius and electric permittivity taken from the solid-state measurements. The calculations show that the second approach gives a better description of the structural modifications during the proton transfer.


Zebraodes, new genus, with a new species from Vietnam (Lepidoptera: Tortricidae: Tortricinae: Archipini).
A new genus of Tortricinae, Zebraodes Heppner & Bae, n. gen., is described for the new species Zebraodes lucidalis Heppner & Bae, n. sp., from Vietnam (Tortricidae: Tortricinae: Archipini). The new genus is anomalous among archipines, resembling some Ceracini, but with genital features more similar to those of Archipini, notably the strongly convex phallus in the male genitalia, together with the strong uncus; however, the absence of a signum in the female genitalia is unusual for the tribe.


Studies on the vitamin D endocrine system in the avian.
This report summarizes the current understanding of the mode of action of vitamin D and emphasizes the contributions to this system that have been based on results obtained in avian species. A complex endocrine system coordinates the metabolism of vitamin D into 1,25-dihydroxycholecalciferol[1,25(OH)2D3] and 24R,25-dihydroxycholecalciferol[24R,25)OH)2D3] and 28 other metabolites; of these 16 were originally isolated and chemically characterized from avian systems. Key advances in understanding the mode of action of the seco-steroid 1,25(OH)2D3 and the scope of its action have been made by studying the tissue distribution of both its receptor and its gene-induced product, a 28,000-dalton calcium-binding protein termed calbindin-D28K. To date no less than 23 tissues have been found to have specific 1,25(OH)2D3 receptors; of these 10 were identified in avian studies. Similarly, nine tissues express the vitamin D-induced calbindin; seven have been reported in avian tissues. The second dihydroxylated metabolite, 24R,25(OH)2D3, has been reported to be capable of inducing a variety of specific biological effects, some when the steroid is administered alone and some in the presence of its companion dihydroxylated metabolite 1,25(OH)2D3. There is emerging evidence for the existence of specific receptors for 24R,25(OH)2D3, particularly in chondrocytes and parathyroid glands (both studied in avian systems). These observations collectively demonstrate the broad scope of the vitamin D endocrine system.


The epidermal growth factor receptor tyrosine kinase inhibitor gefitinib sensitizes colon cancer cells to irinotecan.
Epidermal growth factor receptor (EGFR) overactivity plays a significant role in colon cancer biology and has been associated with poor clinical prognosis. Early clinical trials reported efficacy of receptor-targeted compounds, including modulation of clinical irinotecan resistance. We investigated the effects of the EGFR tyrosine kinase inhibitor gefitinib on cellular determinants of irinotecan resistance in human colon cancer cells. At non-cytotoxic concentrations, gefitinib sensitized colon cancer cells to SN-38, the active metabolite of irinotecan. Gefitinib increased the SN-38-mediated induction of protein-linked DNA single-strand breaks in a dose-dependent manner, with no alteration of topoisomerase (Topo) I protein expression or enzymatic activity. Whereas Topo IIbeta protein expression was not affected by gefitinib, significant time- and concentration-dependent downregulation of Topo IIalpha protein and inhibition of its enzymatic function were observed, corresponding to a G1 phase cell cycle arrest. Gefitinib significantly inhibited EGFR-associated signaling molecules, including phospho-mitogen-activated protein kinase or protein kinase C, which may account for decreases in proliferation or topoisomerase activity, respectively. Although a dose-dependent decrease of the BCRP/MXR/ABCP half-transporter was observed under gefitinib, cellular pharmacokinetics revealed no significant differences in accumulation or retention of the active SN-38 lactone using reverse-phase HPLC analysis. This study delineates mechanisms that may contribute to the synergism observed between irinotecan and EGFR inhibitors.


Antisense and sense RNA probe hybridization to immobilized crude cellular lysates: a tool to screen growth hormone antagonists.
The growth-promoting effect of growth hormone (GH) is primarily mediated by insulin-like growth factor-1 (IGF-1). The liver is the main source of circulating IGF-I. The authors have used rodent primary hepatocytes for studies on pharmacological intervention of IGF-I mRNA expression. A 96-well nonradioactive IGF-1 mRNA quantification assay was developed, based on the hybridization of sense and antisense RNA probes, to replicate membranes with crude hepatocyte lysates. The sense hybridization was used as an internal standard. The antagonistic properties of a set of GH-receptor binding compounds were evaluated. Two compounds were found to down-regulate IGF-I mRNA. Effects due to metabolic inhibition or toxicity were excluded using a cell proliferation assay. To investigate potential unspecific transcriptional effects, the mRNA levels of the housekeeping genes, beta-actin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), were determined. Two other GH-regulated genes, cytochrome P450 2C12 (CYP2C12) and a rat homologue to the human alpha1B-glycoprotein (A1BG), were quantified by RNase protection assays and found to be down-regulated, confirming the antagonistic property of 1 compound. In conclusion, a direct filter hybridization assay of hepatocyte lysates using nonradioactive sense and antisense probes can be used for quantitative mRNA measurements and could constitute a valuable tool in screening for pharmacologically active compounds.


Combined strategy of endothelial cells coating, Sertoli cells coculture and infusion improves vascularization and rejection protection of islet graft.
Improving islet graft revascularization and inhibiting rejection become crucial tasks for prolonging islet graft survival. Endothelial cells (ECs) are the basis of islet vascularization and Sertoli cells (SCs) have the talent to provide nutritional support and exert immunosuppressive effects. We construct a combined strategy of ECs coating in the presence of nutritious and immune factors supplied by SCs in a co-culture system to investigate the effect of vascularization and rejection inhibition for islet graft. In vivo, the combined strategy improved the survival and vascularization as well as inhibited lymphocytes and inflammatory cytokines. In vitro, we found the combinatorial strategy improved the function of islets and the effect of ECs-coating on islets. Combined strategy treated islets revealed higher levels of anti-apoptotic signal molecules (Bcl-2 and HSP-32), survival and function related molecules (PDX-1, Ki-67, ERK1/2 and Akt) and demonstrated increased vascular endothelial growth factor receptor 2 (KDR) and angiogenesis signal molecules (FAk and PLC-γ). SCs effectively inhibited the activation of lymphocyte stimulated by islets and ECs. Predominantly immunosuppressive cytokines could be detected in culture supernatants of the SCs coculture group. These results suggest that ECs-coating and Sertoli cells co-culture or infusion synergistically enhance islet survival and function after transplantation.


Progestin Intrauterine Devices and Metformin: Endometrial Hyperplasia and Early Stage Endometrial Cancer Medical Management.
Globally, endometrial cancer is the sixth leading cause of female cancer-related deaths. Non-atypical endometrial hyperplasia (EH), has a lifetime progression rate to endometrial cancer ranging from less than 5%, if simple without atypia, to 40%, if complex with atypia. Site specific, long-acting intrauterine devices (IUDs) provide fertility sparing, progestin-based EH medical management. It is unclear which IUD is most beneficial, or if progesterone sensitizing metformin offers improved outcomes. For resolution, PubMed searches for "Mirena" or "Metformin," "treatment," "endometrial hyperplasia," or "stage 1 endometrial cancer," were performed, yielding 33 articles. Of these, 19 articles were included. The 60 mg high-dose frameless IUD/20 mcg levonorgestrel has achieved sustained regression of Grade 3 endometrial intraepithelial neoplasia for 14 years. Case series on early stage endometrial cancer (EC) treatment with IUDs have 75% or greater regression rates. For simple through complex EH with atypia, the 52 mg-IUD/10-20 mcg-LNG-14t has achieved 100% complete regression in 6-months. Clearly, IUDs have an outcome advantage over oral progestins. However, studies on metformin for EH, and of progestins or metformin for early stage EC management are underpowered, with inadequate dose ranges to achieve significant differences in, or optimal outcomes for, the treatment modalities. Therefore, outcomes from the feMMe trial for the 52 mg-IUD/10-20 mcg-LNG-14t and metformin will fill a gap in the literature.


Resistant Strains of Enterotoxigenic Staphylococcus aureus; Unknown Risk for Multiple Sclerosis Exacerbation.
Despite all advances in neurological sciences, there are unknown aspects in the epidemiology of multiple sclerosis (MS). Based on this hypothesis, the enterotoxigenic strains of Staphylococcus aureus (S. aureus) are possible risk factors for exacerbations of MS. The present study was carried out to investigate the role of resistant strains of enterotoxigenic S. aureus in MS exacerbation. Two-hundred nasal swab samples were collected from non-MS (n = 80), MS stable (n = 60) and MS exacerbation (n = 60) groups. Samples were cultured and those that were S. aureus-positive were analyzed for the presence of enterotoxins, using polymerase chain reaction (PCR). Antimicrobial susceptibility was performed using disk diffusion method. Ninety out of 200 nasal samples (45%) were positive for S. aureus. The highest levels of nasal colonization were seen in MS exacerbation group (68.33%). The most commonly detected enterotoxins were sea (30%), sec (15.55%) and seb (11.11%). There were significant differences between S. aureus colonization and type of samples (P = 0.026) and, also, between type of samples and prevalence of enterotoxins (P = 0.022). The highest levels of enterotoxigenic genes were seen in MS exacerbation group. The S. aureus strains had the highest levels of resistance against tetracycline (80%), ampicillin (72.22%), methicillin (66.66%), erythromycin (66.66%), oxacillin (63.33%), trimethoprim-sulfamethoxazole (61.11%) and cotrimoxazole (55.55%). Our findings should raise awareness about the role of sea and sec enterotoxins, in resistant strains of S. aureus, as a risk factor for MS exacerbation. It is better to keep MS patients away from polluted environments of hospitals and health centers.


Safety evaluation of clinical gene therapy using hepatocyte growth factor to treat peripheral arterial disease.
Therapeutic angiogenesis using angiogenic growth factors is expected to be a new treatment for patients with critical limb ischemia (CLI). Because hepatocyte growth factor (HGF) has potent angiogenic activity, we investigated the safety and efficiency of HGF plasmid DNA in patients with CLI as a prospective open-labeled clinical trial. Intramuscular injection of naked HGF plasmid DNA was performed in ischemic limbs of 6 CLI patients with arteriosclerosis obliterans (n=3) or Buerger disease (n=3) graded as Fontaine III or IV. The primary end points were safety and improvement of ischemic symptoms at 12 weeks after transfection. Severe complications and adverse effects caused by gene transfer were not detected in any patients. Of particular importance, no apparent edema was observed in any patient throughout the trial. In addition, serum HGF concentration was not changed throughout the therapy period in all patients. In contrast, a reduction of pain scale of more than 1 cm in visual analog pain scale was observed in 5 of 6 patients. Increase in ankle pressure index more than 0.1 was observed in 5 of 5 patients. The long diameter of 8 of 11 ischemic ulcers in 4 patients was reduced >25%. Intramuscular injection of naked HGF plasmid is safe, feasible, and can achieve successful improvement of ischemic limbs. Although the present data are conducted to demonstrate the safety as phase I/early phase IIa, the initial clinical outcome with HGF gene transfer seems to indicate usefulness as sole therapy for CLI.


The complement SC5b-9 complex mediates cell adhesion through a vitronectin receptor.
Adhesion of cells to the terminal complement complex of C5b through C9 containing the serum S-protein (SC5b-9) was investigated using a microtiter plate attachment assay with L8 myoblast indicator cells. The skeletal muscle-derived L8 myoblasts bound and spread on substratum coated with SC5b-9, and with the vitronectin/S-protein component of SC5b-9. The myoblasts did not adhere to substratum coated with collagen, laminin, or fibronectin. The cell attachment was blocked by antibody to vitronectin/S-protein, whereas antibody to the other components C5, C6, C7, C8, or C9 had minimal effect. The cells were not bound to free vitronectin because attachment activity was removed by adsorption with an anti-C6 antibody column. The L8 cell attachment was dependent on divalent cations, was blocked by synthetic peptides containing the amino acid sequence Arg-Gly-Asp, and was inhibited by antivitronectin receptor antibody. These results indicate that cells adhere to the SC5b-9 complex through interaction of the vitronectin component with an integrin vitronectin receptor. Cell attachment to terminal C complexes could be used for leukocyte adherence and migration during inflammation, and also for attachment of tissue cells during regeneration after disease or traumatic injury.


Involvement of CCL2/CCR2 macrophage recruitment in amodiaquine-induced liver injury.
Evidence suggests that macrophages may play a role in the development of idiosyncratic drug-induced liver injury (IDILI). However, there has yet to be a clear link between macrophage activation and the inflammatory infiltrate that is characteristic of IDILI. A major chemokine involved in the recruitment of macrophages into the liver is C-C motif chemokine ligand 2 (CCL2)/monocyte chemoattractant protein 1 (MCP1). Therefore, we tested the effect of this chemokine in an animal model of IDILI. Specifically, amodiaquine (AQ), which is known to cause IDILI in humans, causes mild liver injury in wild-type C57BL/6 mice that resolves despite continued AQ treatment, but it causes more severe liver injury that does not resolve in PD-1-/- mice co-treated with anti-CTLA-4 to impair immune tolerance. CCR2-/- mice treated with AQ were not protected from the expected AQ-induced liver injury seen in wild-type C57BL/6 mice. In contrast, anti-CCL2 antibodies attenuated the liver injury caused by AQ in the impaired immune tolerance model. The difference in response of the two models is likely due to a difference in the IDILI mechanism; the mild injury in wild-type animals is mediated by NK cells, while the more serious injury in the impaired immune tolerance model requires CD8 T-cells. The results from these experiments provide evidence that macrophage infiltration into the liver may not be involved in mild IDILI mediated by the innate immune system, but it does appear necessary in more severe IDILI involving cytotoxic T-cells.


Biotin transport and accumulation by cells of Lactobacillus plantarum. I. General properties of the system.
Waller, James R. (University of Cincinnati, Cincinnati, Ohio), and Herman C. Lichstein. Biotin transport and accumulation by cells of Lactobacillus plantarum. I. General properties of the system. J. Bacteriol. 90:843-852. 1965.-Resting cells of Lactobacillus plantarum were saturated with bound biotin by incubation in phosphate buffer with biotin and glucose for 2 hr. This bound biotin was stable to wide changes in temperature, pH, and reaction time. Free biotin could be eluted from the cells by suspending them in cold water or saline. Immersing the cells in boiling water for 30 sec released all free biotin. Recoveries of added biotin exceeded 92%. Free biotin uptake by bound biotin-saturated cells occurred by two mechanisms. One process was independent from temperature (Q(10), 1.25), pH, cellular metabolism, and inhibition by iodoacetate. The other mechanism was dependent upon temperature (Q(10), 2.58; optimum, 37 C), pH (optimum, 7.5), and active cellular metabolism, and was inhibited by iodoacetate. Activation energies of 3,700 and 13,800 cal per mole, respectively, were observed for glucose-independent and -dependent free biotin uptake. Both processes exhibited approximately the same degree of inhibition by homobiotin. Higher concentrations of homobiotin were required to inhibit growth than to inhibit free biotin uptake. Intracellular-extracellular ratios as high as 600 were established in the absence of glucose, whereas ratios of nearly 4,000 occurred in the presence of glucose.


Thalamic afferents from the brain stem. An experimental study using retrograde single and double labelling with HRP and iron-dextran in the rat. I. Medial and lateral reticular formation.
The reticulo-thalamic projection of the rat was studied in single and double experiments using 51 injections of HRP and/or iron-dextran. The medial reticular formation (excluding the ncl. pedunculo-pontinus, subncl. compactus, see Part II) sends out projections to the ncll. intralaminares (IL), ncl. mediodorsalis (MD), ncl. ventralis medialis (VM), ncl. ventralis lateralis (VL), ncl. ventralis basalis (VB) and ncl. posterior (PO) of the thalamus. The mesencephalic RF (ncl. cuneiformis and ncl. subcuneiformis) projects to all the above mentioned nuclei. The rhombencephalic RF (ncl. pontis oralis, ncl. pontis caudalis, ncl. gigantocellularis et paragigantocellularis dorsalis et lateralis) projects to the posterior IL (ncl. parafascicularis and ncl. centromedianus) and to the MD, pars paralaminaris. The pontine RF (ncl. pontis oralis et caudalis) projects to the posterior IL and to the medial part of the ncl. parafascicularis. The RF of the pons and the upper oblongata projects to the VL, VB and PO. The lateral RF (ncl. parvocellularis) projects solely from the tiers around the ncl. n. VII. to the posterior IL in what is a scanty projection. We found no connections projecting from the medial or lateral RF to the midline nuclei, to the lateral nuclei or to the ncl. commissurae posterioris; the projection to the ncl. reticularis thalami was left unstudied. The reticulo-thalamic pathways exhibit marked laterality. The mesencephalic RF projects to the thalamus in what is pronounced ipsilateral predominance, the pontine RF in ipsilateral predominance, the upper oblongata RF equally ipsi- and contralaterally, and the lower oblongata RF with a predominance of the contralateral component. Collateralization (double labelled cells) was found projecting solely to the IL and MD regions. It was both ipsilateral and contralateral. The discoveries included the topographical organization of the reticulo-thalamic projection: a major antero-posterior in the rhombencephalon and a mediolateral in the projections from the mesencephalon and from the pons. The most substantial projection was found aiming at the posterior IL nuclei (ncl. parafascicularis-centrum medianum), a less substantial one to the ventral and posterior nuclei, and the scantiest to the anterior IL nuclei (ncl. paracentralis, ncl. centralis lateralis).


Phase-engineered high-entropy metastable FCC Cu2-y Ag y (In x Sn1-x )Se2S nanomaterials with high thermoelectric performance.
Crystal-phase engineering to create metastable polymorphs is an effective and powerful way to modulate the physicochemical properties and functions of semiconductor materials, but it has been rarely explored in thermoelectrics due to concerns over thermal stability. Herein, we develop a combined colloidal synthesis and sintering route to prepare nanostructured solids through ligand retention. Nano-scale control over the unconventional cubic-phase is realized in a high-entropy Cu2-y Ag y (In x Sn1-x )Se2S (x = 0-0.25, y = 0, 0.07, 0.13) system by surface-ligand protection and size-driven phase stabilization. Different from the common monoclinic phase, the unconventional cubic-phase samples can optimize electrical and thermal properties through phase and entropy design. A high power factor (0.44 mW m-1 K-2), an ultralow thermal conductivity (0.25 W m-1 K-1) and a ZT value of 1.52 are achieved at 873 K for the cubic Cu1.87Ag0.13(In0.06Sn0.94)Se2S nanostructured sample. This study highlights a new method for the synthesis of metastable phase high-entropy materials and gives insights into stabilizing the metastable phase through ligand retention in other research communities.


Color removal from distillery spent wash through coagulation using Moringa oleifera seeds: use of optimum response surface methodology.
The effects of dosage, pH and concentration of salts were investigated for an optimized condition of color removal from the distillery spent wash. The optimization process was analyzed using custom response surface methodology (RSM). The design was employed to derive a statistical model for the effect of parameters studied on removal of color using Moringa oleifera coagulant (MOC). The dosage (20 and 60 ml), pH (7 and 8.5) and concentration of 0.25 M had been found to be the optimum conditions for maximum 56% and 67% color removal using sodium chloride (NaCl) and potassium chloride (KCl) salts respectively. The actual color removal at optimal conditions was found to be 53% and 64% respectively for NaCl and KCl salts which confirms close to RSM results. The effects of storage duration and temperature on MOC studied reveal that coagulation efficiency of MOC kept at room temperature was effective for 3 days and at 4 degrees C it performed coagulation up to 5 days.


The sequence-dependent unfolding pathway plays a critical role in the amyloidogenicity of transthyretin.
Human transthyretin (TTR) is an amyloidogenic protein whose aggregation is associated with several types of amyloid diseases. The following mechanism of TTR amyloid formation has been proposed. TTR tetramer at first dissociates into native monomers, which is the rate-limiting step in fibril formation. The monomeric species then partially unfold to form amyloidogenic intermediates that subsequently undergo a downhill self-assembly process. The amyloid deposit can be facilitated by disease-associated point mutations. However, only subtle structural differences were observed between the crystal structures of the wild type and the disease-associated variants. To investigate how single-point mutations influence the effective energy landscapes of TTR monomers, molecular dynamics (MD) simulations were performed on wild-type TTR and two pathogenic variants. Principal coordinate analysis on MD-generated ensembles has revealed multiple unfolding pathways for each protein. Amyloidogenic intermediates with the dislocated C strand-loop-D strand motif were observed only on the unfolding pathways of V30M and L55P variants and not for wild-type TTR. Our study suggests that the sequence-dependent unfolding pathway plays a crucial role in the amyloidogenicity of TTR. Analyses of side chain concerted motions indicate that pathogenic mutations on "edge strands" disrupt the delicate side chain correlated motions, which in turn may alter the sequence of unfolding events.


Binding of thrombin to thrombomodulin accelerates inhibition of the enzyme by antithrombin III. Evidence for a heparin-independent mechanism.
The endothelial cell surface provides a receptor for thrombin-designated thrombomodulin (TM) which regulates thrombin formation and the activity of the enzyme at the vessel wall surface by serving as a potent cofactor for the activation of protein C by thrombin. Heparin-like structures of the vessel wall have been proposed as another regulatory mechanism catalyzing the inhibition of thrombin by antithrombin III. In the present study, the interaction of antithrombin III with the thrombin-TM complex and its interference with heparin and polycations were investigated by using human components and TM isolated from the microvasculature of rabbit lung. Purified TM bound thrombin and acted as a cofactor for protein C activation. The addition of heparin (0.5 unit/mL) to the reaction mixture interfered neither with the binding of thrombin to TM nor with the activation of protein C. However, the polycations protamine (1 unit/mL) as well as polybrene (0.1 mg/mL) affected the thrombin-TM interaction. This was documented by an increase in the Michaelis constant from 8.3 microM for thrombin alone to 19.5 microM for thrombin-TM with the chromogenic substrate compound S-2238 in the presence of 1 unit/mL protamine. When the inhibition of thrombin by antithrombin III was determined, the second-order rate constant k2 = 8.4 X 10(3) M-1 s-1 increased about 8-fold in the presence of TM, implying an accelerative function of TM in this reaction. Although purified TM did not bind to antithrombin III-Sepharose, suggesting the absence of heparin-like structures within the receptor molecule, protamine reversed the accelerative effect of TM in the inhibition reaction.(ABSTRACT TRUNCATED AT 250 WORDS)


Body potassium content and 40K radiation dose to Iranian subjects.
In this study 215 Iranian subjects including 180 males and 35 females were investigated for potassium body contents, its variations with age, weight, body build index, and slenderness, as well as the radiation dose due to 40K. The average total body potassium is 147 +/- 22 g for males and 100 +/- 16 g for females. The average potassium concentrations for males and females are 1.98 +/- 0.34 and 1.60 +/- 0.29 g kg(-1) body weight, respectively. The potassium content decreases with age for both sexes. The 40K concentration decreases with slenderness in both sexes, particularly in females. The total body potassium increases with body build index for males, while its value is constant for females less than 40 y, and increases for females more than 40 y. The average effective doses due to 40K are 187 +/- 32 microSv y(-1) for males and 150 +/- 27 microSv y(-1) for females.


Probing OH stretching overtones of CH3OOH through action spectroscopy: Influence of dipole moment dependence on HOOC torsion.
The OH stretching overtones and OH stretch-HOOC torsion combination bands of methyl hydroperoxide are investigated using action spectroscopy initiated through vibrational state selected photodissociation. Our results for the room temperature spectra covering the 2nu(OH)-5nu(OH) regions suggest that the coarse vibrational structures appearing in the spectra can be understood using a simple two-dimensional vibration-torsion model involving the OH stretch and COOH torsion consistent with what has been previously reported. However, investigation of the jet-cooled spectrum for the 2nu(OH) band along with the results of ab initio calculations using coupled cluster methods reveals that the dependence of the transition dipole moment on the HOOC torsion angle cannot be neglected when simulating intensities of OH stretching overtone bands, as has been suggested by earlier room temperature studies. The present results demonstrate that transitions between torsional levels of different symmetries, which arise from the dependence of the dipole moment mu(r,tau) on the torsional angle, contribute significantly to the intensities of the vibrational overtone bands and are important in interpreting the temperature dependence of the spectral band profiles. Contributions from these transitions are largest for the 2nu(OH) and 3nu(OH) levels and fall off gradually for the higher overtones. In addition, results are presented investigating the orbital interactions in CH(3)OOH that influence changes in the HOOC adiabatic torsion potential with increased OH stretching excitation.


The nucleotide sequence of spinach chloroplast tryptophan transfer RNA.
Spinach chloroplast tRNATrp, purified by column chromatography and two-dimensional gel electrophoresis, has been sequenced using in vitro labeling techniques. The sequence is : pG-C-G-C-U-C-U-U-A-G-U-U-C-A-G-U-U-C-Gm-G-D-A-G-A-A-C-m2G-psi-G-G-G-psi-C-U-C-A-A*-A-A-C-C-C-G-A-U-G-N-C-G-U-A-G-G-T-psi-C-A-A-G-U-C-C-U-A-C-A-G-A-G-C-G-U-G -C-C-AOH. Like the E. coli suppressor tRNA psu+UGA which translates both the opal terminator codon U-G-A and the tryptophan codon U-G-G, spinach chloroplast tRNATrp has C-C-A as an anticodon and contains an A-U pair in the D-stem.


[Radiologic diagnosis of early complications following kidney transplantation].
Postoperative complications after renal transplantation, such as hematomas, abscesses, urinomas, lymphoceles and obstruction of the ureter, can be diagnosed by means of sonography. Early vascular complications, such as venous thrombosis and arterial occlusion or stenosis, can be recognized by duplex sonography and accurately depicted by angiography to allow planning of surgical intervention. Stenosis of the renal artery is amenable to treatment by percutaneous angioplasty. Parenchymatous complications (acute tubular necrosis, acute rejection, cyclosporin-A-toxicity) can cause changes in the intrarenal flow patterns at duplex sonography, but this examination does not allow accurate diagnosis and differentiation of these changes. Magnetic resonance imaging is a very promising method for the differential diagnosis of parenchymatous complications.


Induction of subterahertz surface waves on a metal wire by intense laser interaction with a foil.
We have demonstrated that a pulsed electromagnetic wave (Sommerfeld wave) of subterahertz frequency and 11-MV/m field strength can be induced on a metal wire by the interaction of an intense femtosecond laser pule with an adjacent metal foil at a laser intensity of 8.5×10^{18}W/cm^{2}. The polarity of the electric field of this surface wave is opposite to that obtained by the direct interaction of the laser with the wire. Numerical simulations suggest that an electromagnetic wave associated with electron emission from the foil induces the surface wave. A tungsten wire is placed normal to an aluminum foil with a gap so that the wire is not irradiated and damaged by the laser pulse, thus making it possible to generate surface waves on the wire repeatedly.


Extracting the Auditory Attention in a Dual-Speaker Scenario From EEG Using a Joint CNN-LSTM Model.
Human brain performs remarkably well in segregating a particular speaker from interfering ones in a multispeaker scenario. We can quantitatively evaluate the segregation capability by modeling a relationship between the speech signals present in an auditory scene, and the listener's cortical signals measured using electroencephalography (EEG). This has opened up avenues to integrate neuro-feedback into hearing aids where the device can infer user's attention and enhance the attended speaker. Commonly used algorithms to infer the auditory attention are based on linear systems theory where cues such as speech envelopes are mapped on to the EEG signals. Here, we present a joint convolutional neural network (CNN)-long short-term memory (LSTM) model to infer the auditory attention. Our joint CNN-LSTM model takes the EEG signals and the spectrogram of the multiple speakers as inputs and classifies the attention to one of the speakers. We evaluated the reliability of our network using three different datasets comprising of 61 subjects, where each subject undertook a dual-speaker experiment. The three datasets analyzed corresponded to speech stimuli presented in three different languages namely German, Danish, and Dutch. Using the proposed joint CNN-LSTM model, we obtained a median decoding accuracy of 77.2% at a trial duration of 3 s. Furthermore, we evaluated the amount of sparsity that the model can tolerate by means of magnitude pruning and found a tolerance of up to 50% sparsity without substantial loss of decoding accuracy.


Improving outcomes and mitigating costs associated with CAR T-cell therapy.
Since the historic approval of tisagenlecleucel for the treatment of B-cell acute lymphoblastic leukemia in 2017, chimeric antigen receptor (CAR) T-cell therapies have altered the treatment paradigm for hematologic malignancies. Five CAR T-cell products are now approved by the US Food and Drug Administration for a growing number of cancer indications and a global market worth billions is anticipated in the next 5 years. While uptake of CAR T-cell therapy is rapidly ramping up, there remain significant barriers to effective implementation and patient access, not least their price tag and substantial ancillary costs of care. CAR T-cell therapies currently have the potential to be cost-effective; however, improved safety and efficacy, outpatient administration, and a streamlined manufacturing process could make them even more so. In the meantime, payers and providers are tasked with facing the logistical complexities of CAR T-cell therapy and developing new payment and reimbursement strategies to ensure value-based care and optimal access today.


Detection of cranial meningiomas: comparison of ⁶⁸Ga-DOTATOC PET/CT and contrast-enhanced MRI.
PET imaging with somatostatin receptor ligands, such as (68)Ga-DOTATOC, is a well-established method for detection and target volume definition of meningiomas prior to radiotherapy. Since DOTATOC PET delivers a higher contrast between meningiomas and surrounding tissues than MRI, we conducted a retrospective analysis to compare the diagnostic accuracy of contrast-enhanced MRI (CE-MRI) with (68)Ga-DOTATOC PET/CT in patients with cranial meningiomas prior to radiotherapy. Over a period of 6 years, 134 patients (20-82 years of age, 107 women and 27 men) underwent cranial CE-MRI and (68)Ga-DOTATOC PET/CT. To compare the two methods, the lesions considered typical of meningiomas visually were counted and analysed with respect to their location and SUVmax. In the 134 patients investigated by both modalities, 190 meningiomas were detected by (68)Ga-DOTATOC PET/CT and 171 by CE-MRI. With knowledge of the PET/CT data, the MRI scans were reinvestigated, which led to the detection of 4 of the 19 incidental meningiomas, resulting in an overall detection rate of 92 % of the meningioma lesions that were found by PET/CT. Ga-DOTATOC PET/CT demonstrated an improved sensitivity in meningioma detection when compared to CE-MRI. Tumours adjacent to the falx cerebri, located at the skull base or obscured by imaging artefacts or calcification are particularly difficult to detect by MRI. Therefore (68)Ga-DOTATOC PET/CT may provide additional information in patients with uncertain or equivocal results on MRI or could help to confirm a diagnosis of meningioma based on MRI or could help to confirm MRI-based diagnosis of meningiomas in cases of biopsy limitations. It is possible that not only radiotherapy and surgical planning, but also follow-up strategies would benefit from this imaging modality.


ERCC1 mRNA expression is associated with the clinical outcome of non-small cell lung cancer treated with platinum-based chemotherapy.
We conducted a prospective study to analyze the expression of the excision repair cross-complementing group 1 (ERCC1) and ribonucleotide reductase subunit M1 (RRM1) genes in 297 Chinese patients with advanced non-small cell lung cancer (NSCLC). The goal of this study was to evaluate these genes as potential biomarkers for prediction of tumor response and clinical outcome. Patients with unresectable, locally advanced or metastatic NSCLC were enrolled between September 2007 and September 2009, and they were followed up until September 2012. A fluorescence-based real-time detection method was used to quantify relative levels of ERCC1 and RRM1 cDNA. Relative amounts of ERCC1 and RRM1 cDNA were calculated by comparing to actin. By the end of follow-up, 132 patients had died and 165 patients experienced progression. The median overall survival time was 18.7 months (range, 1-60 months). The median levels of ERCC1 and RRM1 were 2.46 x 10(-2) and 0.97 x 10(-2), respectively. Patients with low ERCC1 expression had a significantly higher rate of complete response to chemotherapy, with an OR (95%CI) of 1.56 (1.03-2.47). Moreover, individuals with low levels of ERCC1 had longer overall survival than patients with high expression, with an adjusted hazard ratio (95%CI) of 0.57 (0.35-0.93). In summary, low ERCC1 mRNA expression was associated with better response to chemotherapy and correlated with longer survival in advanced NSCLC patients treated with platinum-based chemotherapy.


Do doctors undertreat pain?
Routinely, physicians discount patients' pain reports and provide too little analgesia too late. Critics call them callous, sadistic, and Puritanical, but the causes of these clinical pratices are different -- namely, a psychological need to distance themselves from the pain they encounter and inflict, and more subtly, a peculiar concept of pain acquired in medical training. Physicians learn to think of pain as a symptom to observe and explore in diagnosing and monitoring disease -- not as a complaint to relieve quickly or fully. Moreover, pain-relief is regarded as subordinate to, and competing with, efforts to cure or maintain the life of a patient. This training, I suggest, gives physicians a new, clinical concept of pain at odds with their prior, lay concept of pain whose manifestations standardly call for sympathetic efforts at relief. The conceptual nature of this difference is obscured by thinking of pain as a solely private sensation, rather than as a sensation with public and social aspects (à la Wittgenstein). Although suppressed in certain clinical circumstances, these standard public and social aspects are shown in the very tests used in clinical pain research. This clinical pain concept is rooted in Medicine conceived as preeminently curative and life-prolonging. Physicians are, however, themselves undermining this professional self-definition (by treating AIDS and Alzheimer's patients; by no longer pressing their patients to 'fight to the end'; by collaborating with non-medical healers). Accordingly, pain-relief may gain greater therapeutic status, and, so too, the ordinary concept of pain that medical training has suppressed.


[SOCIAL-ECONOMIC-DEMOGRAPHIC DIFFERENCES IN REASONS NOT TO COMPLY IN TIME WITH ROUTINE CHILDHOOD VACCINATIONS].
In recent years, there has been an increase in parents who do not comply with recommended routine vaccines. The vaccination coverage rates in Israel are known, but the reasons for not completing immunizations are unknown. The aim of this study was to assess the extent of failure to complete the routine vaccinations in time and the reasons among different population groups. This is a retrospective case-control cohort study, among children born in 2009. A total of 14,232 medical records of children who received services at Mother-Child Health Clinics were checked. The children who received all three core vaccines (3th Hepatitis B, 4th DTaP and MMR vaccines) were considered fully compliant and those missing at least one vaccination were considered non-compliant. Six reasons for not complying were identified: medical reasons, parental refusal to vaccinate, requests for a deferred schedule of vaccinations, parental behavior, organizational reasons and others. The differences between groups were analyzed by univariate analysis and logistic regression. In this study, 7.4% of children, were not fully compliant with all three core vaccines within two years. The main reasons in the Jewish population were: refusal and requests for variation in scheduling (44.3%) and in the Arabic population: parental behavior (40.8%). The risk for not completing the vaccinations was higher in children of Jewish mothers, residence in the Hadera sub-district, parents who had an academic education and older parents. There are differences between Arab and Jewish populations in the reasons for not complying in time with childhood vaccinations. In order to obtain herd immunity in the Israeli population, we need to tailor interventions suitable for each population group according to their characteristics.


Patient-Centered Radiology Communications: Engaging Patients as Partners.
Patient-centered care is a model in which, by bringing the patient's perspective to the design and delivery of health care, we can better meet patents' needs, enhancing the quality of care. Patient-centered care requires finding ways to communicate effectively with a diverse patient population that has various levels of health literacy, cultural backgrounds, and unique needs and preferences. Moreover, multimedia resources have the potential to inform and educate patients promoting greater independence. In this review, we discuss the fundamentals of communication with the different modes used in radiology and the key elements of effective communication. Then, we highlight five opportunities along the continuum of care in the radiology practice in which we can improve communications to empower our patients and families and strengthen this partnership. Lastly, we discuss the importance on communication training of the workforce, optimizing and seamlessly integrating technology solutions into our workflows, and the need for patient feedback in the design and delivery of care.


The inverse problem of acoustic wave scattering by an air-saturated poroelastic cylinder.
The efficient use of plastic foams in a diverse range of structural applications like in noise reduction, cushioning, and sleeping mattresses requires detailed characterization of their permeability and deformation (load-bearing) behavior. The elastic moduli and airflow resistance properties of foams are often measured using two separate techniques, one employing mechanical vibration methods and the other, flow rates of fluids based on fluid mechanics technology, respectively. A multi-parameter inverse acoustic scattering problem to recover airflow resistivity (AR) and mechanical properties of an air-saturated foam cylinder is solved. A wave-fluid saturated poroelastic structure interaction model based on the modified Biot theory and plane-wave decomposition using orthogonal cylindrical functions is employed to solve the inverse problem. The solutions to the inverse problem are obtained by constructing the objective functional given by the total square of the difference between predictions from the model and scattered acoustic field data acquired in an anechoic chamber. The value of the recovered AR is in good agreement with that of a slab sample cut from the cylinder and characterized using a method employing low frequency transmitted and reflected acoustic waves in a long waveguide developed by Fellah et al. [Rev. Sci. Instrum. 78(11), 114902 (2007)].


A New MI-Based Visualization Aided Validation Index for Mining Big Longitudinal Web Trial Data.
Web-delivered clinical trials generate big complex data. To help untangle the heterogeneity of treatment effects, unsupervised learning methods have been widely applied. However, identifying valid patterns is a priority but challenging issue for these methods. This paper, built upon our previous research on multiple imputation (MI)-based fuzzy clustering and validation, proposes a new MI-based Visualization-aided validation index (MIVOOS) to determine the optimal number of clusters for big incomplete longitudinal Web-trial data with inflated zeros. Different from a recently developed fuzzy clustering validation index, MIVOOS uses a more suitable overlap and separation measures for Web-trial data but does not depend on the choice of fuzzifiers as the widely used Xie and Beni (XB) index. Through optimizing the view angles of 3-D projections using Sammon mapping, the optimal 2-D projection-guided MIVOOS is obtained to better visualize and verify the patterns in conjunction with trajectory patterns. Compared with XB and VOS, our newly proposed MIVOOS shows its robustness in validating big Web-trial data under different missing data mechanisms using real and simulated Web-trial data.