DTPACE: an effective, novel combination chemotherapy with thalidomide for previously treated patients with myeloma.
To improve outcome in previously treated patients (at least two cycles of standard therapy) with multiple myeloma, thalidomide was combined with cytotoxic chemotherapy as induction therapy. The regimen consisted of 4-days of oral dexamethasone, daily thalidomide, and 4 days of continuous-infusion cisplatin, doxorubicin, cyclophosphamide, and etoposide (DTPACE). Response to two cycles of DTPACE for induction was evaluated in 236 patients. Before being treated with DTPACE, 148 patients (63%) had shown progressive disease while receiving standard chemotherapy, and 55 patients (23%) had chromosome 13 abnormalities. The partial remission rate (PR) after two cycles of DTPACE was 32%, with 16% attaining a complete remission (CR) or near-CR (nCR; defined as only immunofixation electrophoresis-positive). Patients with high lactate dehydrogenase (LDH; n = 98) showed a better response than those with normal LDH (n = 138): PR or better, 43% v 27% (P =.01); CR + nCR, 25% v 11% (P =.01). Patients with chromosome 13 abnormalities (n = 55) responded equally well as the other patients (n = 181): PR or better, 35% v 33% (P =.84); CR + nCR, 17% v 15% (P =.73). Patients who received 100% dose of DTPACE for two cycles (n = 115) achieved higher response rates than those with less than 100% dose (n = 121): PR or better, 49% v 17% (P <.0001); CR + nCR, 27% v 6% (P <.0001). Combination therapy of oral dexamethasone and thalidomide with infusional chemotherapy is effective as induction therapy before autotransplantation, especially in patients with high-risk features.


New validated high-performance liquid chromatographic method for simultaneous analysis of ten flavonoid aglycones in plant extracts using a C18 fused-core column and acetonitrile-tetrahydrofuran gradient.
An HPLC method of high resolution has been developed and validated for the simultaneous determination of ten prominent flavonoid aglycones in plant materials using a fused-core C18-silica column (Ascentis® Express, 4.6 mm × 150 mm, 2.7 μm). The separation was accomplished with an acetonitrile-tetrahydrofuran gradient elution at a flow rate of 1 mL/min and temperature of 30°C. UV spectrophotometric detection was employed at 370 nm for flavonols (quercetin [QU], myricetin [MY], isorhamnetin [IS], kaempferol [KA], sexangularetin [SX], and limocitrin [LM]) and 340 nm for flavones (apigenin [AP], acacetin [AC], chrysoeriol [CH], and luteolin [LU]). The high resolution of critical pairs QU/LU (10.50), QU/CH (3.40), AP/CH (2.51), SX/LM (2.30), and IS/KA (2.70) was achieved within 30.3 min. The observed column back pressure was less than 4300 psi, thus acceptable for conventional HPLC equipment. The method was sensitive enough having LODs of 0.115-0.525 ng and good linearity (r > 0.9999) over the test range. The precision values, expressed as RSD values, were <7.5%, and the accuracy was in the range of 95.3-100.2% for all analytes except MY (73.8%). The method was successfully employed for the determination of flavonoids in several medicinal plants, such as Ginkgo biloba, Betula pendula, and a variety of Sorbus species.


Parkinson's disease and risk of mortality: meta-analysis and systematic review.
To evaluate the existing prospective observational studies on the morality risk among Parkinson's disease (PD) patients and determine the overall risk ratio (RR) of mortality by conducting a meta-analysis and systematic review. Original articles published in English were searched in PubMed and Embase databases prior to March 2013. Only prospective observational studies providing adjusted risk estimates related to PD and future mortality were considered eligible. Pooled adjusted RR and 95% confidence interval (CI) were computed either by fixed-effects models or by random-effects models. Eight studies with 72,833 participants were identified and analysed. In the pooled analyses, patients with PD had a greater risk of all-cause mortality (RR = 2.22; 95% CI: 1.78-2.77). Subgroup analyses based on the design, gender, follow-up duration and sample size showed that a consistent positive association between PD and the mortality risk in each subgroup. However, no statistical significance was found for the baseline age <65 years (RR = 1.42; 95% CI: 0.72-2.77). PD patients with dementia had particularly high mortality risks (RR = 3.78; 95% CI: 2.06-6.92). This meta-analysis indicated that among patients with PD, the all-cause mortality increased by 2.22-fold compared with the general population. PD patients with dementia particularly had higher risks of mortality.


Affinity labeling of mu opioid receptors by sulfhydryl alkylating derivatives of morphine and morphinone.
After reduction of a disulfide bond at or near the mu opioid binding site in rat brain membranes, incubating membranes with 14 beta-bromoacetamido derivatives of either morphine, dihydromorphine, morphinone, or dihydromorphinone resulted in the irreversible inhibition of mu opioid binding to rat brain membranes. Without the addition of the disulfide bond-reducing reagent dithiothreitol, these affinity ligands bound reversibly to opioid binding sites. Binding to either delta or kappa opioid binding sites was not altered by alkylation of the membranes with the affinity ligands. The percentage of irreversible inhibition of mu opioid binding was dependent on the time and temperature of the incubation of membranes with the affinity ligands and on the concentrations of dithiothreitol and the affinity ligands. Incubating membranes with morphine afforded almost complete protection from alkylation of the mu opioid binding site. Naloxone and the l-isomer levorphanol also protected the site from alkylation, whereas the d-isomer dextrorphan and the kappa-selective opioid U50,488H did not protect the site. The mu-selective peptide [D-Ala2, (Me)Phe4,Gly(ol)5]enkephalin was the peptide that afforded the greatest protection. These studies have shown that, after the reduction of a disulfide bond at or near the mu opioid binding site, this sulfhydryl group can be specifically alkylated, resulting in the affinity labeling of the mu opioid binding site.


Cytomegalovirus glycoprotein B sequence variation among Japanese bone marrow transplant recipients.
Glycoprotein B (gB) of human cytomegalovirus (CMV) is the major target protein of neutralizing antibodies, and four variant types are known. A previously reported polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method was modified by eliminating the cell culture step and amplifying a CMV fragment (corresponding to a.a. 441-450 of gB) by nested PCR. DNA was extracted from the sera of 27 pediatric bone marrow transplant recipients and subjected to nested PCR. Of the samples, 20 yielded PCR products, and the gB type was determined by RFLP. Of the 20 patients, 4 (20%) had gB type 1 (Towne type), 15 patients (75%) had gB type 2 (AD 169 type), and 1 patient (5%) had gB type 3. Previous studies showed that gB type was most common among Caucasians [Chou and Dennison: J Infect Dis 1991;163:1229-1234; Fries et al: J Infect Dis 1994;169:769-774]. Thus, gB genotypes seem to be distributed differently in Caucasians and Japanese. Further, nucleotide sequence analysis of the amplified region revealed that all the type 2 viruses had the same amino acid sequences. The type 3 sample had a novel amino acid substitution at position 498. Of the type 1 samples, 3 had amino acid substitutions in various positions: 1 sample at position 493, 1 sample at position 447 and 1 sample at position 452, 493 and 498.


Abrupt climate change at the end of the last glacial period inferred from trapped air in polar Ice
The last glacial period was terminated by an abrupt warming event in the North Atlantic approximately 15,000 years before the present, and warming events of similar age have been reported from low latitudes. Understanding the mechanism of this termination requires that the precise relative timing of abrupt climate warming in the tropics versus the North Atlantic be known. Nitrogen and argon isotopes in trapped air in Greenland ice show that the Greenland Summit warmed 9 +/- 3 degrees C over a period of several decades, beginning 14,672 years ago. Atmospheric methane concentrations rose abruptly over a approximately 50-year period and began their increase 20 to 30 years after the onset of the abrupt Greenland warming. These data suggest that tropical climate became warmer or wetter (or both) approximately 20 to 80 years after the onset of Greenland warming, supporting a North Atlantic rather than a tropical trigger for the climate event.


Uncertainty and risk in wildland fire management: a review.
Wildland fire management is subject to manifold sources of uncertainty. Beyond the unpredictability of wildfire behavior, uncertainty stems from inaccurate/missing data, limited resource value measures to guide prioritization across fires and resources at risk, and an incomplete scientific understanding of ecological response to fire, of fire behavior response to treatments, and of spatiotemporal dynamics involving disturbance regimes and climate change. This work attempts to systematically align sources of uncertainty with the most appropriate decision support methodologies, in order to facilitate cost-effective, risk-based wildfire planning efforts. We review the state of wildfire risk assessment and management, with a specific focus on uncertainties challenging implementation of integrated risk assessments that consider a suite of human and ecological values. Recent advances in wildfire simulation and geospatial mapping of highly valued resources have enabled robust risk-based analyses to inform planning across a variety of scales, although improvements are needed in fire behavior and ignition occurrence models. A key remaining challenge is a better characterization of non-market resources at risk, both in terms of their response to fire and how society values those resources. Our findings echo earlier literature identifying wildfire effects analysis and value uncertainty as the primary challenges to integrated wildfire risk assessment and wildfire management. We stress the importance of identifying and characterizing uncertainties in order to better quantify and manage them. Leveraging the most appropriate decision support tools can facilitate wildfire risk assessment and ideally improve decision-making.


Work function analysis of vegetarian entrée production.
Data on labor time for food production can be used as an effective management tool. It is essential for foodservice managers to know how labor time is being used (1). A continuous time study was conducted to determine total labor time for the production of eight vegetarian entrées in a hospital foodservice system. Two work areas were observed: the ingredient assembly area and the cooks' production area. Times were recorded by work function to identify how labor time was distributed. Results showed (a) observed frequency for each work function, (b) time expended in seconds per portion for each work function, (c) percentage distribution of labor time by work function, (d) total time for each employee involved in entrée production, and (e) percentage of total time in which each employee was involved in the production of each entrée. Total labor time varied by type of entrée, ranging from 39.97 to 19.33 seconds per portion. Entrées with the highest labor time required the largest amount of hand labor. A one-way analysis of variance indicated significant differences in mean labor time among the eight vegetarian entrées for direct labor time (p = .0009), and total labor time (p = .0018). No significant differences were found among entrées for indirect labor or delay time.


A simple gas chromatography method for the analysis of monoethanolamine in air.
A simple method determining airborne monoethanolamine has been developed. Monoethanolamine determination has traditionally been difficult due to analytical separation problems. Even in recent sophisticated methods, this difficulty remains as the major issue often resulting in time-consuming sample preparations. Impregnated glass fiber filters were used for sampling. Desorption of monoethanolamine was followed by capillary GC analysis and nitrogen phosphorous selective detection. Separation was achieved using a specific column for monoethanolamines (35% diphenyl and 65% dimethyl polysiloxane). The internal standard was quinoline. Derivatization steps were not needed. The calibration range was 0.5-80 μg/mL with a good correlation (R(2) = 0.996). Averaged overall precisions and accuracies were 4.8% and -7.8% for intraday (n = 30), and 10.5% and -5.9% for interday (n = 72). Mean recovery from spiked filters was 92.8% for the intraday variation, and 94.1% for the interday variation. Monoethanolamine on stored spiked filters was stable for at least 4 weeks at 5°C. This newly developed method was used among professional cleaners and air concentrations (n = 4) were 0.42 and 0.17 mg/m(3) for personal and 0.23 and 0.43 mg/m(3) for stationary measurements. The monoethanolamine air concentration method described here was simple, sensitive, and convenient both in terms of sampling and analytical analysis.


Recombinant p35 from bacteria can form Interleukin (IL-)12, but Not IL-35.
The Interleukin (IL)-12 family contains several heterodimeric composite cytokines which share subunits among each other. IL-12 consists of the subunits p40 (shared with IL-23) and p35. p35 is shared with the composite cytokine IL-35 which comprises of the p35/EBI3 heterodimer (EBI3 shared with IL-27). IL-35 signals via homo- or heterodimers of IL-12Rβ2, gp130 and WSX-1, which are shared with IL-12 and IL-27 receptor complexes, respectively. p35 was efficiently secreted in complex with p40 as IL-12 but not with EBI3 as IL-35 in several transfected cell lines tested which complicates the analysis of IL-35 signal transduction. p35 and p40 but not p35 and EBI3 form an inter-chain disulfide bridge. Mutation of the responsible cysteine residue (p40C197A) reduced IL-12 formation and activity only slightly. Importantly, the p40C197A mutation prevented the formation of antagonistic p40 homodimers which enabled the in vitro reconstitution of biologically active IL-12 with p35 produced in bacteria (p35bac). Reconstitution of IL-35 with p35bac and EBI3 did, however, fail to induce signal transduction in Ba/F3 cells expressing IL-12Rβ2 and gp130. In summary, we describe the in vitro reconstitution of IL-12, but fail to produce recombinant IL-35 by this novel approach.


Is there a place for preimplantation genetic diagnosis screening in recurrent miscarriage patients?
Chromosomal abnormalities are one of the factors known to interfere with normal embryo development; thus preimplantation genetic diagnosis (PGD) for chromosome anomalies may be a new tool for improving the pregnancy rate in selected groups of patients. Embryos from three groups of patients (control, aged and recurrent miscarriage patients) were screened by PGD using specific DNA probes for chromosomes 13, 16, 18, 21, 22, X and Y. The control and aged groups were included in the PGD study because the women carried sex-linked genetic disease. The frequencies of chromosome anomalies observed in older women (46.3%) and in recurrent miscarriage patients (53%) were significantly higher (P < 0.05) than the frequency in the control group (19.3%). After PGD screening and transfer of normal embryos, pregnancies were obtained in women who had undergone repeated abortions (approximate 25% pregnancy rate per transfer) but not in older women. On the basis of these data, it seems that PGD screening of embryos can help some women undergoing repeated abortions, as these techniques allow successful pregnancies to be obtained or, if no pregnancy can be obtained, the results may help the couple to decide whether to enter an embryo or gamete donation programme.


Guidelines for mouse and human DC functional assays.
This article is part of the Dendritic Cell Guidelines article series, which provides a collection of state-of-the-art protocols for the preparation, phenotype analysis by flow cytometry, generation, fluorescence microscopy, and functional characterization of mouse and human dendritic cells (DC) from lymphoid organs and various non-lymphoid tissues. Recent studies have provided evidence for an increasing number of phenotypically distinct conventional DC (cDC) subsets that on one hand exhibit a certain functional plasticity, but on the other hand are characterized by their tissue- and context-dependent functional specialization. Here, we describe a selection of assays for the functional characterization of mouse and human cDC. The first two protocols illustrate analysis of cDC endocytosis and metabolism, followed by guidelines for transcriptomic and proteomic characterization of cDC populations. Then, a larger group of assays describes the characterization of cDC migration in vitro, ex vivo, and in vivo. The final guidelines measure cDC inflammasome and antigen (cross)-presentation activity. While all protocols were written by experienced scientists who routinely use them in their work, this article was also peer-reviewed by leading experts and approved by all co-authors, making it an essential resource for basic and clinical DC immunologists.


Pediatric laparoscopic splenectomy.
Lateral laparoscopic splenectomy in adults, first reported in 1991, was begun with children in 1993. The authors reviewed records of 59 patients 2 to 17 years old who underwent laparoscopic splenectomy by the lateral approach between 1994 and 1998 at four medical centers. Patients received prophylactic penicillin or vaccinations preoperatively. Of the 59 patients, 51 required splenectomy for one of the following conditions: idiopathic thrombocytopenic purpura, hereditary spherocytosis, or sickle-cell disease. Splenomegaly was found in 86% of the patients, and ten accessory spleens were resected. No deaths or infection occurred, and only three patients had perioperative complications: acute chest crisis, small diaphragmatic injury, and intraoperative hemorrhage. One operation was converted to a minilaparatomy because of difficulty with specimen extraction. Pediatric laparoscopic splenectomy is safe and effective, resulting in little blood loss, rapid recovery, and a good cosmetic outcome.


Culture, intangibles and metrics in environmental management.
The demand for better representation of cultural considerations in environmental management is increasingly evident. As two cases in point, ecosystem service approaches increasingly include cultural services, and resource planners recognize indigenous constituents and the cultural knowledge they hold as key to good environmental management. Accordingly, collaborations between anthropologists, planners, decision makers and biodiversity experts about the subject of culture are increasingly common-but also commonly fraught. Those whose expertise is culture often engage in such collaborations because they worry a practitioner from 'elsewhere' will employ a 'measure of culture' that is poorly or naively conceived. Those from an economic or biophysical training must grapple with the intangible properties of culture as they intersect with economic, biological or other material measures. This paper seeks to assist those who engage in collaborations to characterize cultural benefits or impacts relevant to decision-making in three ways; by: (i) considering the likely mindset of would-be collaborators; (ii) providing examples of tested approaches that might enable innovation; and (iii) characterizing the kinds of obstacles that are in principle solvable through methodological alternatives. We accomplish these tasks in part by examining three cases wherein culture was a critical variable in environmental decision making: risk management in New Zealand associated with Māori concerns about genetically modified organisms; cultural services to assist marine planning in coastal British Columbia; and a decision-making process involving a local First Nation about water flows in a regulated river in western Canada. We examine how 'culture' came to be manifest in each case, drawing from ethnographic and cultural-models interviews and using subjective metrics (recommended by theories of judgment and decision making) to express cultural concerns. We conclude that the characterization of cultural benefits and impacts is least amenable to methodological solution when prevailing cultural worldviews contain elements fundamentally at odds with efforts to quantify benefits/impacts, but that even in such cases some improvements are achievable if decision-makers are flexible regarding processes for consultation with community members and how quantification is structured.


Comparison of unidimensional and bidimensional measurements in metastatic non-small cell lung cancer.
Tumour response evaluation after chemotherapy has become crucial in the development of many drugs. In contrast to the standard bidimensional WHO criteria, the recently described Response Evaluation Criteria In Solid Tumors are based on unidimensional measurements. The aim of the present study was to compare both methods in patients with metastatic non-small cell lung cancer. One hundred and sixty-four patients treated with two cisplatin-paclitaxel-based chemotherapy schedules between June 1994 and December 2000 were analysed. The measurements were reviewed by an independent panel of radiologists. Patient characteristics were: median age of 55 years (range 24-77 years) and a male to female ratio of 129 : 35. Adenocarcinoma and squamous carcinoma were the most common histologies. Vinorelbine was the third drug used in 77 patients and gemcitabine in 87. The ratio unidimensional/bidimensional was as follows: response 85 : 85; stable disease 32 : 32; progression 47 : 42 and not assessable 0 : 5. Kappa for agreement between responders was 0.951 (95% CI: 0.795-1.0) (P<0.001). Both WHO criteria and Response Evaluation Criteria In Solid Tumors give similar results in assessing tumour response in patients with non-small cell lung cancer after chemotherapy. The unidimensional measurement could replace the more complex bidimensional one.


A UNIFIED FRAMEWORK FOR ESTIMATING DIFFUSION TENSORS OF ANY ORDER WITH SYMMETRIC POSITIVE-DEFINITE CONSTRAINTS.
Cartesian tensors of various orders have been employed for either modeling the diffusivity or the orientation distribution function in Diffusion-Weighted MRI datasets. In both cases, the estimated tensors have to be positive-definite since they model positive-valued functions. In this paper we present a novel unified framework for estimating positive-definite tensors of any order, in contrast to the existing methods in literature, which are either order-specific or fail to handle the positive-definite property. The proposed framework employs a homogeneous polynomial parametrization that covers the full space of any order positive-definite tensors and explicitly imposes the positive-definite constraint on the estimated tensors. We show that this parametrization leads to a linear system that is solved using the non-negative least squares technique. The framework is demonstrated using synthetic and real data from an excised rat hippocampus.


The role of luteinizing hormone in human follicle development and oocyte fertility: evidence from in-vitro fertilization in a woman with long-standing hypogonadotrophic hypogonadism and using recombinant human follicle stimulating hormone.
To evaluate the relative importance of follicle stimulating hormone (FSH) and luteinizing hormone (LH) in follicular development and oocyte fertility in the human species, the use of recombinant human FSH, human menopausal gonadotrophin (HMG), and very highly purified urinary human FSH (FSH-HP) plus oestradiol valerate for ovarian stimulation and in-vitro fertilization (IVF) were compared in three cycles in a woman with isolated congenital gonadotrophin deficiency who had never been treated with ovarian stimulating agents. The total number of ampoules of gonadotrophins used was lower in the HMG treatment cycle. Ovarian response and IVF outcome in the three treatment cycles were as follows: (i) HMG cycle: normal follicular growth, normal pattern of oestradiol and inhibin through the menstrual cycle, high fertilization rate (93%); (ii) recombinant FSH cycle: normal follicular growth, low oestradiol and abnormal inhibin, finally poor rate of fertilization (28%); (iii) FSH-HP plus oestradiol valerate cycle: normal follicular growth, normal pattern of inhibin and poor fertilization rate (27%). Luteal plasma progesterone concentrations were much higher in the HMG treatment cycle. This case shows that FSH is the only factor required in order to induce follicular growth in the human, although LH or a product derived from its action may assist in order to achieve full follicular maturity and oocytes capable of fertilization. Though oestradiol might have a mediatory role in the process of follicular maturation, our results favour a direct primary role of LH in complete maturation of the follicle.


Pet-1 Controls Tetrahydrobiopterin Pathway and Slc22a3 Transporter Genes in Serotonin Neurons.
Coordinated serotonin (5-HT) synthesis and reuptake depends on coexpression of Tph2, Aadc (Ddc), and Sert (Slc6a4) in brain 5-HT neurons. However, other gene products play critical roles in brain 5-HT synthesis and transport. For example, 5-HT synthesis depends on coexpression of genes encoding the enzymatic machinery necessary for the production and regeneration of tetrahydrobiopterin (BH4). In addition, the organic cation transporter 3 (Oct3, Slc22a3) functions as a low affinity, high capacity 5-HT reuptake protein in 5-HT neurons. The regulatory strategies controlling BH4 and Oct3 gene expression in 5-HT neurons have not been investigated. Our previous studies showed that Pet-1 is a critical transcription factor in a regulatory program that controls coexpression of Tph2, Aadc, and Sert in 5-HT neurons. Here, we investigate whether a common regulatory program determines global 5-HT synthesis and reuptake through coordinate transcriptional control. We show with comparative microarray profiling of flow sorted YFP(+) Pet-1(-/-) and wild type 5-HT neurons that Pet-1 regulates BH4 pathway genes, Gch1, Gchfr, and Qdpr. Thus, Pet-1 coordinates expression of all rate-limiting enzymatic (Tph2, Gch1) and post-translational regulatory (Gchfr) steps that determine the level of mammalian brain 5-HT synthesis. Moreover, Pet-1 globally controls acquisition of 5-HT reuptake in dorsal raphe 5-HT neurons by coordinating expression of Slc6a4 and Slc22a3. In situ hybridizations revealed that virtually all 5-HT neurons in the dorsal raphe depend on Pet-1 for Slc22a3 expression; similar results were obtained for Htr1a. Therefore, few if any 5-HT neurons in the dorsal raphe are resistant to loss of Pet-1 for their full neuron-type identity.


5-fluorouracil, leucovorin, and irinotecan (FOLFIRI) as a third-line chemotherapy treatment in metastatic gastric cancer, after failure of fluoropyrimidine, platinum, anthracycline, and taxane.
Studies on the effects of third-line chemotherapy (CT) in advanced gastric cancer (GC) patients are still scarce. The aim of this study was to evaluate the efficacy and safety of the modified 5-fluorouracil, leucovorin, and irinotecan (mFOLFIRI) regimen as a third-line CT in metastatic GC patients, after failure of fluoropyrimidine, platinum, anthracycline, and taxane. After failure of first- and second-line therapies, 42 patients received third-line FOLFIRI (180 mg/m² irinotecan and 400 mg/m² leucovorin administered concomitantly as a 90-minute intravenous (IV) infusion on day 1, followed by a 400 mg/m² 5-fluorouracil IV bolus then 2600 mg/m² continuous infusion over 46 hours), between January 2009 and December 2015. FOLFIRI was administered for a median of 6 cycles (range 4-12 cycles). Eight patients achieved partial response, while 13 patients showed stable disease, resulting in the overall response rate (ORR) of 19% and disease control rate (DCR) of 50%. The most frequent grade 3-4 hematological and non-hematological toxicities were neutropenia (14.2%) and diarrhea (7.1%). The median progression-free survival (PFS) and overall survival (OS) from the start of third-line CT were 3.8 months (95% confidence interval [CI], 3.0-4.5) and 6.8 months (95% CI, 5.6-7.9), respectively. According to the multivariate analysis, two factors were independently predictive of the poor OS: >2 regions of metastasis (relative risk [RR], 2.6; 95% CI, 1.3-5.4) and a high level of carcinoembryonic antigen [CEA] (RR, 3.4; 95% CI, 1.6-7.4). In conclusion, FOLFIRI was well tolerated as third-line CT and showed promising PFS and OS in advanced GC patients, after failure of fluoropyrimidine, platinum, anthracycline, and taxane.


Environment Changes Genetic Effects on Respiratory Conditions and Allergic Phenotypes.
The prevalence of asthma and allergic diseases is disproportionately distributed among different populations, with an increasing trend observed in Western countries. Here we investigated how the environment affected genotype-phenotype association in a genetically homogeneous, but geographically separated population. We evaluated 18 single nucleotide polymorphisms (SNPs) corresponding to 8 genes (ADAM33, ALOX5, LT-α, LTC4S, NOS1, ORMDL3, TBXA2R and TNF-α), the lung function and five respiratory/allergic conditions (ever asthma, bronchitis, rhinitis, dermatitis and atopy) in two populations of Inuit residing either in the westernized environment of Denmark or in the rural area of Greenland. Our results showed that lung function was associated with genetic variants in ORMDL3, with polymorphisms having a significant interaction with place of residence. LT-α SNP rs909253 and rs1041981 were significantly associated with bronchitis risk. LT-α SNP rs2844484 was related to dermatitis susceptibility and was significantly influenced by the place of residence. The observed gene-phenotype relationships were exclusively present in one population and absent in the other population. We conclude that the genotype-phenotype associations relating to bronchitis and allergy susceptibility are dependent on the environment and that environmental factors/lifestyles modify genetic predisposition and change the genetic effects on diseases.


Successful use of chronic epoprostenol as a bridge to liver transplantation in severe portopulmonary hypertension.
Portopulmonary hypertension, defined as mean pulmonary artery pressure >25 mmHg in the presence of a normal pulmonary capillary wedge pressure and portal hypertension, is a known complication of end-stage liver disease that has been associated with high morbidity and mortality at the time of liver transplantation. We have recently reported the successful treatment of portopulmonary hypertension with chronic intravenous epoprostenol and now report the first patient with severe portopulmonary hypertension successfully treated with epoprostenol who subsequently underwent successful liver transplantation. A patient with severe portopulmonary hypertension was treated with intravenous epoprostenol, 23 ng/kg/min, for a 4-month period, after which the portopulmonary hypertension resolved and the patient underwent successful liver transplantation. The patient was discharged, continues to do well, and at 3 months is off epoprostenol with near normal pulmonary artery pressures. Chronic epoprostenol, in conjunction with a multidisciplinary, well-planned perioperative evaluation and treatment plan, may be the answer to a heretofore untreatable disease.


An optimal power-dispatching system using neural networks for the electrochemical process of zinc depending on varying prices of electricity.
Depending on varying prices of electricity, an optimal power-dispatching system (OPDS) is developed to minimize the cost of power consumption in the electrochemical process of zinc (EPZ). Due to the complexity of the EPZ, the main factors influencing the power consumption are determined by qualitative analysis, and a series of conditional experiments is conducted to acquire sufficient data, then two backpropagation neural networks are used to describe these relationships quantitatively. An equivalent Hopfield neural network is constructed to solve the optimization problem where a penalty function is introduced into the network energy function so as to meet the equality constraints, and inequality constraints are removed by alteration of the Sigmoid function. This OPDS was put into service in a smeltery in 1998. The cost of power consumption has decreased significantly, the total electrical energy consumption is reduced, and it is also beneficial to balancing the load of the power grid. The actual results show the effectiveness of the OPDS. This paper introduces a successful industrial application and mainly presents how to utilize neural networks to solve particular problems for the real world.


The binding of inhibitors to alpha-chymotrypsin at alkaline pH.
1. The binding of the competitive inhibitor N-acetyl-d-tryptophan amide to alpha-chymotrypsin has now been studied at pH values up to 10.6, by the technique of equilibrium dialysis. 2. This binding depends on the ionization of a group on the free enzyme with apparent pK(a) 9.3 at 5 degrees . 3. This group is tentatively identified as that responsible for an enzyme conformation change at high pH values, on which the catalytic activity of the enzyme also depends.


Treatment of tick-bite fever with erythromycin.
Because of the occasional need for an alternative drug for the treatment of tick-bite fever, a study of the value of erythromycin for the variety of tick-borne typhus fever occurring in southern Africa was undertaken. In guinea-pigs erythromycin 125 mg/kg for 5 days largely prevented the fever and other signs of the infection, but did not prevent the serological reaction as determined by the rickettsial complement fixation test. A series of 17 human patients were treated for 4 days with erythromycin 500 mg 6-hourly for adults and 30-50 mg/kg/d in 4 divided doses for children. Eleven patients appeared to respond favourably. It was concluded that erythromycin does not have as specific a value as tetracycline in the treatment of tick-bite fever, but that when tetracycline is contraindicated erythromycin is a useful alternative drug.


Shikonin induces apoptosis and G0/G1 phase arrest of gallbladder cancer cells via the JNK signaling pathway.
Shikonin, a natural product isolated from the roots of Lithospermum erythrorhizon, is considered to have antitumor effects. Gallbladder cancer (GBC) is a prevalent biliary tract malignancy with few curative therapeutic stragegies and poor prognosis. In the present study, we detected the effects of shikonin on GBC cells as well as the underlying molecular mechanisms. The results demonstrated that GBC cell proliferation was inhibited by shikonin as determined by MTT and colony formation assays. Flow cytometry results demonstrated that shikonin treatment enhanced apoptosis and promoted G0/G1 phase arrest in the GBC cells. Western blot assay showed that shikonin induced mitochondrial-dependent apoptosis via the JNK signaling pathway. Moreover, shikonin suppressed tumor growth in mice bearing GBC-derived xenografts in a dose‑dependent manner without side-effects. These results revealed that shikonin exhibits anticancer effects on GBC cells by inducing apoptosis and regulating the cell cycle. Taken together, shikonin may be a novel and safe chemotherapeutic agent for the treatment of GBC.


Recognition and referral of depression in patients with heart disease.
Routine screening and assessment for depression occurs rarely in clinical practice for a variety of reasons, including the absence of systems to facilitate the process. To identify nurses' knowledge and practice regarding depression screening and referral for cardiac patients following the implementation of education workshops and a validated screening tool with referral actions. Pre and post-test design using surveys and semi-structured interviews was conducted with a purposive sample of nurses in a large Australian metropolitan tertiary referral hospital. Prior to the introduction of the screening and referral tool, nurses engaged in an interactive one hour education workshop on the topic of depression and the tool; introduced to improve depression screening following an acute cardiac event. In the pre and post-survey 40 and 30 nurses, respectively, participated with 14 also engaging in semi-structured interviews. Eighty percent reported a 'good' understanding of depression post-program compared to 30% at baseline. Sixty percent reported routinely using the depression screening and referral tool. The interviews identified three main themes that supported the utility of the education and instrument: knowledge improvement; perceived self-efficacy and new knowledge into practice. The vast majority of participants reported increased skill, knowledge and confidence to screen and refer for depression post an acute cardiac event. The substantial increase in the number of nurses who engaged in screening and referral actions further demonstrated the success of the program. These encouraging results provide evidence that screening for depression can be achieved through adopting formalised processes.


IL-19 Contributes to the Development of Nonalcoholic Steatohepatitis by Altering Lipid Metabolism.
Interleukin (IL)-19, a member of the IL-10 family, is an anti-inflammatory cytokine produced primarily by macrophages. Nonalcoholic steatohepatitis (NASH) is a disease that has progressed from nonalcoholic fatty liver disease (NAFLD) and is characterized by inflammation and fibrosis. We evaluated the functions of IL-19 in a NAFLD/NASH mouse model using a 60% high fat diet with 0.1% methionine, without choline, and with 2% cholesterol (CDAHFD). Wild-type (WT) and IL-19 gene-deficient (KO) mice were fed a CDAHFD or standard diet for 9 weeks. Liver injury, inflammation, and fibrosis induced by CDAHFD were significantly worse in IL-19 KO mice than in WT mice. IL-6, TNF-α, and TGF-β were significantly higher in IL-19 KO mice than in WT mice. As a mechanism using an in vitro experiment, palmitate-induced triglyceride and cholesterol contents were decreased by the addition of IL-19 in HepG2 cells. Furthermore, addition of IL-19 decreased the expression of fatty acid synthesis-related enzymes and increased ATP content in HepG2 cells. The action of IL-19 in vitro suppressed lipid metabolism. In conclusion, IL-19 may play an important role in the development of steatosis and fibrosis by directly regulating liver metabolism and may be a potential target for the treatment of liver diseases.


Describing congenital heart disease by using three-part segmental notation.
The segmental classification of congenital heart disease, first conceptualized nearly 50 years ago, is now well established. The Van Praagh classification system, in particular, is commonly used throughout North America to facilitate communication between physicians from various specialties who are involved in diagnosing and managing congenital cardiovascular abnormalities. In the Van Praagh system, a three-part notation consisting of letters separated by commas and encompassed by a set of braces is used to succinctly describe the visceroatrial situs, the orientation of the ventricular loop, and the position and relation of the great vessels. For example, the notation "{S, D, S}" describes the normal anatomic configuration, in which the morphologic right atrium and largest hepatic lobe are on the patient's right side and the morphologic left atrium, stomach, and spleen are on the left side (situs solitus); the ventricular loop is curved rightward (dextro- or d-loop); and the aorta is posterior to and rightward of the main pulmonary artery (situs solitus). Because the Van Praagh notation imposes on its users a systematic approach to anatomic description, it is a helpful device for structuring the interpretation of imaging studies as well as the reporting of results: First, the morphologic right and left atria and ventricles must be identified; next, the visceroatrial situs and ventricular loop orientation may be determined from the positions of the cardiac chambers; and last, the position and relation of the great vessels (normal, inverted, transposed, or malpositioned) can be established. The article provides concise, step-by-step guidance for applying the Van Praagh system in the radiology reading room.


Phase II study of 2-chlorodeoxyadenosine in combination with chlorambucil in previously untreated B-cell chronic lymphocytic leukemia.
The objective was to determine the safety and efficacy of adding a maximally tolerated dose of 2-chlorodeoxyadenosine (2-CdA) to standard chlorambucil (CLB) therapy in previously untreated B-cell chronic lymphocytic leukemia (CLL). Thirty patients with CLL (median age, 64 years) received two courses of 2-CdA given intravenously (2 mg/m2 daily for 7 days) added to biweekly administration of CLB at 30 mg/m2 given orally. The diagnosis of CLL, treatment indications, and response criteria were according to the National Cancer Institute established guidelines. Sixteen patients (53%) had advanced-stage disease, and four (13%) had trisomy 12 abnormality. The overall remission rate was 80%, including 20% complete remission (CR), 30% nodular partial remission (nPR), and 30% partial remission (PR). Minimal residual disease was detected phenotypically in two of five patients with CR and in eight of nine with nPR. Overall, CR, nPR, and PR rates were not influenced significantly by the presence of cytogenetic abnormalities or advanced clinical stage. With a median follow-up of 33 months, 58% of patients who had a response had relapse. Median time to progression in all 30 patients was 30 months, and time to progression and progression-free survival were not significantly different for the different response groups, clinical stages, or cytogenetic groups. Severe neutropenia and thrombocytopenia occurred in 33% and 7% of patients, respectively. Only two patients had documented bacterial infections, and four had herpetic infections. Concurrent combination chemotherapy with abbreviated doses of 2-CdA and standard-dose CLB is feasible and safe in previously untreated CLL. Antitumor activity may be superior to that of CLB alone given in conventional doses. Whether a different schedule of combining these two agents would result in improved outcome is being investigated.


Molecular evolution of bat color vision genes.
The two suborders of bats, Megachiroptera (megabats) and Microchiroptera (microbats), use different sensory modalities for perceiving their environment. Megabats are crepuscular and rely on a well-developed eyes and visual pathway, whereas microbats occupy a nocturnal niche and use acoustic orientation or echolocation more than vision as the major means of perceiving their environment. In view of the differences associated with their sensory systems, we decided to investigate the function and evolution of color vision (opsin genes) in these two suborders of bats. The middle/long wavelength (M/L) and short wavelength (S) opsin genes were sequenced from two frugivorous species of megabats, Haplonycteris fischeri and Pteropus dasymallus formosus, and one insectivorous species of microbat, Myotis velifer. Contrary to the situation in primates, where many nocturnal species have lost the functional S opsin gene, both crepuscular and strictly nocturnal species of bats that we examined have functional M/L and S opsin genes. Surprisingly, the S opsin in these bats may be sensitive to UV light, which is relatively more abundant at dawn and at dusk. The M/L opsin in these bats appears to be the L type, which is sensitive to red and may be helpful for identifying fruits among leaves or for other purposes. Most interestingly, H. fischeri has a recent duplication of the M/L opsin gene, representing to date the only known case of opsin gene duplication in non-primate mammals. Some of these observations are unexpected and may provide insights into the effect of nocturnal life on the evolution of opsin genes in mammals and the evolution of the life history traits of bats in general.


Estimating the accuracy of pharmacophore-based detection of cognate receptor-ligand pairs in the immunoglobulin superfamily.
Secreted and membrane-bound members of the immunoglobulin superfamily (IgSF) encompass a large, diverse array of proteins that play central roles in immune response and neural development, and are implicated in diseases ranging from cancer to rheumatoid arthritis. Despite the potential biomedical benefits of understanding IgSF:IgSF cognate receptor-ligand interactions, relatively little about them is known at a molecular level, and experimentally probing all possible receptor-ligand pairs is prohibitively costly. The Protein Ligand Interface Design (ProtLID) algorithm is a computational pharmacophore-based approach to identify cognate receptor-ligand pairs that was recently validated in a pilot study on a small set of IgSF complexes. Although ProtLID has shown a success rate of 61% at identifying at least one cognate ligand for a given receptor, it currently lacks any form of confidence measure that can prioritize individual receptor-ligand predictions to pursue experimentally. In this study, we expanded the application of ProtLID to cover all IgSF complexes with available structural data. In addition, we introduced an approach to estimate the confidence of predictions made by ProtLID based on a statistical analysis of how the ProtLID-constructed pharmacophore matches the structures of candidate ligands. The confidence score combines the physicochemical compatibility, spatial consistency, and mathematical skewness of the distribution of matches throughout a set of candidate ligands. Our results suggest that a subset of cases meeting stringent confidence criteria will always have at least one successful receptor-ligand prediction.


Studies on polyethylene glycol-monoclonal antibody conjugates for fabrication of nanoparticles for biomedical applications.
The objective of this work is to synthesize and characterize PEGylated monoclonal antibody using the reactivity of oligosaccharide residues in the Fc region of trastuzumab and pertuzumab with a view to preserving their activities. The hydrazide-functionalized PEG monomethacrylate was synthesized and reacted with NaIO4-generated aldehyde groups on glycans in the Fc-domain of trastuzumab and pertuzumab. The conjugates were purified by HPLC. SAMSA-fluorescein substitution method and MALDI MS spectroscopy were used to determine the number of PEG per antibody. Preliminary biological studies involved antiproliferative studies and binding (flow cytometry) following treatments with SKBR3 (HER2-overexpressing) cells and the control. 1H NMR and 13C NMR confirmed the formation of hydrazide-functionalized PEG monomethacrylate. MALDI mass-spectrometry showed that there are two PEGs per each antibody and it appears more reliable than the degree of SAMSA-fluorescein substitution method. HER-2 binding assay showed that PEGylated monoclonal antibody bound less efficiently to SKBR3 (high HER-2 expressing) cells than unmodified trastuzumab and pertuzumab. In vitro growth inhibitory effects of unmodified monoclonal antibodies increased with increase in concentration; while the in vitro growth inhibitory effects of PEGylated monoclonal antibodies also increased (but less than the pure antibody) with concentration and it appeared to be more active than unmodified mAbs at higher concentration. The results indicate that PEG can be site-specifically attached via the oxidized glycans in the Fc domain of monoclonal antibodies but the process needs further optimization in terms of PEG size and biological testing at each stage of development.


[Locoregional radiotherapy after mastectomy not useful for all Dutch breast cancer patients].
Two recent randomized clinical trials from Denmark and Canada show that postoperative radiotherapy of breast cancer patients in whom the findings revealed a high risk of dying from the disease not only causes improvement of the locoregional tumour control but also prolongs survival. However, this does not establish definitely which patients belong to this 'high risk category'. Results of retrospective studies from Leiden suggest that selective withholding of postoperative radiotherapy in the Netherlands is justified for large groups of breast cancer patients. Both clinically manifest parasternal recurrences and axillary recurrences are rare among these patients. The condition is, however, that the axillary dissection is radical and performed properly.


Maternal and paternal contribution to intergenerational psychosocial transmission of paan chewing.
Paan chewing is a recognized risk factor for oral cancer in the Asian population. However, there is currently little evidence about the intergenerational psychosocial transmission of paan chewing in South Indian families. We investigated the association between parental and participant's paan chewing in a South Indian population. A subset of data was drawn from a hospital-based case-control study on oral cancer, the HeNCe Life study, conducted at Government Dental and Medical Colleges of Kozhikode, South India. Analyses were based on 371 noncancer control participants having diseases unrelated to known risk factors for oral cancer. Demographics, behavioral habits (e.g., paan chewing, smoking), and indicators of socioeconomic position (SEP) of both participants and their parents were collected with the use of a questionnaire-based interview and a life grid technique. Unconditional logistic regression assessed odds ratios (OR) and 95% confidence intervals (95% CI) for the associations between parental and participant's paan chewing, adjusted for confounders. Over half of the participants were males (55.2%), and the mean age of participants was 59 (SD = 12) years. After adjusting for age, religion, parents' SEP, parents' education, smoking and alcohol consumption, and perceived parenting behavior, we observed that maternal paan chewing and paternal paan chewing were significantly associated with the participant's paan chewing ([OR = 2.40, 95% CI = 1.11-5.21] and [OR = 3.05, 95% CI = 1.48-6.27], respectively). Intergenerational psychosocial transmission of the habit of paan chewing could occur through shared sociocultural or environmental factors.


Cytokine mRNA expression in the liver of patients with primary biliary cirrhosis (PBC) and chronic hepatitis B (CHB).
The expression of cytokine mRNA species was determined in liver biopsies from six normal subjects, 18 patients with PBC and 14 patients with hepatitis B e antigen (HBeAg)-positive CHB using a reverse transcriptase-polymerase chain reaction (RT-PCR) technique. cDNA, obtained by reverse transcription using oligo d(T) primers, was amplified by PCR using primers specific for the coding region of seven different cytokines (IL-1, IL-2, IL-4, IL-5, IL-6, interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha)). The abundance of some cytokines (IL-2, IL-4, IL-5 and IFN-gamma) was also estimated by semiquantitative RT-PCR, using as standards dilutions of synthetic cytokine mRNA transcripts, that could be distinguished electrophoretically from respective native cytokine mRNAs. Hepatic inflammation was assessed by a semiquantitative histologic score and by amplification of mRNA for T cell receptor (TCR)-alpha. mRNAs for IL-1 and IL-6 were detected in only one control liver. In CHB, mRNAs for IL-1, IL-2, IL-4, IL-5 and IFN-gamma were detected in 43%, 60%, 80%, 20%, and 54% of biopsies, respectively. mRNA for IFN-gamma and IL-4, but not IL-1, tended to be associated with severe inflammation. In five biopsies semiquantitative analyses revealed increased levels of mRNA for TCR-alpha and, when transcripts were detectable, high levels of mRNA for IFN-gamma and IL-4. In PBC, mRNA for IFN-gamma was detected in 60% of biopsies, but no mRNAs for IL-1, IL-2, IL-4, IL-5, or IL-6, or for TNF-alpha, were detected. Semiquantitative analyses revealed that absolute levels of mRNA for IFN-gamma tended to correlate with the severity of hepatic inflammation. The results suggest that: (i) there may be fundamental differences in the roles that cytokines play in the hepatic inflammatory processes of PBC and CHB; and (ii) while hepatic IFN-gamma mRNA expression is not specific for PBC, IFN-gamma may play a prominent role in the immunopathogenesis of PBC.


Targeted expression of a conditional oncogene in hematopoietic cells of transgenic mice.
We have produced two lines of transgenic mice in which the expression of temperature-sensitive SV-40 large T antigen is targeted to bone marrow megakaryocytes via the platelet factor 4 (PF4) tissue-specific promoter. The progeny of these transgenic mice were observed for about 3 mo, and no malignancies were detected over this period of time. The offspring of these transgenic mice, 6- to 12-wk of age, served as a source of bone marrow cells, which upon in vitro cultivation at the permissive temperature yielded immortalized cell lines (MegT). At the permissive temperature, MegT cells exhibit the characteristics of early 2N and 4N megakaryocytes which include the presence of specific gene products such as PF4, glycoprotein IIb, acetylcholinesterase, and CD45 as well as the absence of molecular markers of other cell lineages such as the macrophage marker Mac-1, the T helper cell marker CD4, the mast cell marker IgE, the T cell marker CD2 or the erythroid cell marker alpha-globin. The inactivation of the oncogene by a shift of temperature from 34 degrees to 39.5 degrees C produces a reduction in the frequency of the 2N cells, in conjunction with the appearance of 8N and 16N cells, consisting of 27 and 3% of total cells, respectively. Thus, we have generated hematopoietic cell lines that are trapped in the early stages of megakaryocyte commitment, but able to undergo part of the normal program of terminal differentiation.


Estimated exposure to bisphenol A in breastfed and breastfed plus formula-fed infants in Turkey: a comparison study.
This study aimed to estimate and compare dietary exposure to bisphenol A (BPA) in exclusively breastfed (EBF) and breastfed plus formula-fed (BF + FF) infants. A total of 70 mothers and their 0-6 month-old infants (40 in the EBF group and 30 in BF + FF group) were included in the study. After the questionnaire form was applied to the mothers, maternal breast milk, infant formula, and infant urine were collected from mother-infant dyads. Total BPA levels in breast milk, infant formula, and infant urine samples were analyzed by the high-pressure liquid chromatography (HPLC). While BPA was detected in 92.5% of the breast milk samples in the EBF group (mean ± SD = 0.59 ± 0.29 ng/mL), BPA was detected in all of the breast milk samples in the BF + FF group (mean ± SD= 0.72 ± 0.37 ng/mL) (p < 0.05). Similarly, 100% of the infant formula samples in the BF + FF group had detectable levels of BPA (mean ± SD = 7.54 ± 1.77 ng/g formula). The mean urinary BPA levels in the EBF infants (4.33 ± 1.89 µg/g creatinine) were not statistically different from the BF + FF infants (5.81 ± 0.11 µg/g creatinine) (p > 0.05). The average daily BPA intake in EBF infants (0.18 ± 0.13 µg/kg body weight (bw)/day) was found to be significantly higher than in BF + FF infants (0.12 ± 0.09 µg/kg bw/day) (p < 0.05). The estimated dietary intakes of BPA for infants in both groups were below the temporary tolerable daily intake (t-TDI) (4 µg/kg bw/day). Consequently, BPA intake of EBF and BF + FF infants were within safe daily limits during the first six months of life.


Enhanced survival in locoregionally confined anaplastic thyroid carcinoma: a single-institution experience using aggressive multimodal therapy.
Historical outcomes in anaplastic thyroid carcinoma (ATC) are poor, with a median survival of only 5 months and <20% of patients surviving 1 year from diagnosis. We hypothesized that survival in newly diagnosed patients with stages IVA and IVB locoregionally confined ATC might be improved by utilizing an aggressive therapeutic approach, prioritizing both the eradication of disease in the neck and preemptive treatment of occult metastatic disease. Between January 1, 2003, and December 31, 2007, 25 new ATC patients were evaluated at our institution. Of these 25 patients, 10 (40%) had metastatic disease at diagnosis and therefore underwent palliative treatment, whereas 5 (20%) had regionally confined disease and desired treatment at their local medical facilities. The remaining 10 consecutive patients (40%) had regionally confined ATC and elected aggressive therapy combining individualized surgery (where feasible), intensity-modulated radiation therapy (IMRT), and radiosensitizing + adjuvant chemotherapy intending four cycles of docetaxel + doxorubicin. Outcomes were assessed on an intention to treat basis. There were no deaths from therapy, but hospitalization was required in two patients (20%) because of treatment-related adverse events. Five patients (50%) are alive and cancer-free, all having been followed >32 months (range: 32-89 months; median: 44 months) with a median overall Kaplan-Meier survival of 60 months. Overall survival at 1 and 2 years was 70% and 60%, respectively, compared to <20% historical survival at 1 year in analogous patients previously treated with surgery and conventional postoperative radiation at our and other institutions. Although based upon a small series of consecutively treated patients, an aggressive approach combining IMRT and radiosensitizing plus adjuvant chemotherapy appears to improve outcomes, including survival in stages IVA and IVB regionally confined ATC, but remains of uncertain benefit in patients with stage IVC (metastatic) disease. Also uncertain is the optimal chemotherapy regimen to use in conjunction with IMRT. Further multicenter randomized trials are required to define optimal therapy in this rare but deadly cancer.


Cell-wall dyes interfere with Cryptococcus neoformans melanin deposition.
Melanization is an intrinsic characteristic of many fungal species, but details of this process are poorly understood because melanins are notoriously difficult pigments to study. While studying the binding of cell-wall dyes, Eosin Y or Uvitex, to melanized and non-melanized Cryptococcus neoformans cells we noted that melanization leads to reduced fluorescence intensity, suggesting that melanin interfered with dye binding to the cell wall. The growth of C. neoformans in melanizing conditions with either of the cell-wall dyes resulted in an increase in supernatant-associated melanin, consistent with blockage of melanin attachment to the cell wall. This effect provided the opportunity to characterize melanin released into culture supernatants. Released melanin particles appeared mostly as networked structures having dimensions consistent with previously described extracellular vesicles. Hence, dye binding to the cell wall created conditions that resembled the 'leaky melanin' phenotype described for certain cell-wall mutants. In agreement with earlier studies on fungal melanins biosynthesis, our observations are supportive of a model whereby C. neoformans melanization proceeds by the attachment of melanin nanoparticles to the cell wall through chitin, chitosan, and various glucans.


Filling of microcirculation in skeletal muscles during timed India ink perfusion.
Lower leg muscles of anesthetized rabbits were perfused in situ with heparinized India ink at flows and pressures comparable to normal resting levels of 4 ml.min-1.100 g-1). Paired cross sections were counterstained with eosin to show ink-containing microvessels and reacted for alkaline phosphate to show all vessels. The fraction of microvessels filled with ink (Fi) increased progressively with perfusion time. At 3.5 s, mean Fi for the muscles studied fell between 0.12 and 0.19. At 60-90 s, the following levels were reached: medial gastrocnemius 0.74, lateral gastrocnemius 0.76, tibialis anterior 0.59, and soleus 0.80. The number of open capillaries and their distribution of flow velocities can be inferred from such data only if the perfusion rate is known or by recourse to a specified anatomic model. The time course of ink appearance shows best agreement with 60-80% of the vessels open and accessible to ink, with microvascular transit times ranging from less than 3 to greater than 30 s. If microvascular path lengths are assumed to be uniform, the range of velocities must be four times to one-fourth the mean, with 15-30% of the microvessels perfused at velocities equal to or greater than the mean. Alternatively, if microvascular velocity is assumed uniform, flow path lengths must vary from one-fourth to four times the average. Anatomic measurements of other suggest that less than one-half the variability in ink transit is attributable to differences in microvascular length. Thus both length and velocity must vary among alternate arteriovenous pathways.


Association of serum creatinine with abnormal hemodynamics and mortality in pulmonary arterial hypertension.
Renal dysfunction predicts mortality in patients with cardiovascular disease. How renal dysfunction relates to hemodynamics and mortality in pulmonary arterial hypertension (PAH) remains unclear. We performed a cohort study of 500 patients with World Health Organization group I PAH from 1982 to 2006 with data on demographics, comorbidities, medications, functional class, laboratory tests, exercise testing results, and hemodynamics. Serum creatinine (SCr) was determined on entry into the study (initial PAH clinic visit). Vital status was determined from hospital records and the Social Security Death Index. We used a Cox proportional hazards analysis to determine whether SCr was an independent predictor of mortality. Mean age on entry into the study was 48+/-14 years, and 79% of subjects were female. Mean SCr was 1.05+/-0.35 mg/dL. Elevated SCr was associated with higher right atrial pressure and lower cardiac index. During a median follow-up of 3.5 years, 279 deaths (55.8% of the cohort) occurred. Compared with patients with SCr <1.0 mg/dL, those with SCr 1.0 to 1.4 mg/dL and SCr >1.4 mg/dL had an increased hazard ratio of death (unadjusted hazard ratio 1.65, 95% confidence interval 1.26 to 2.17, P<0.0001 for SCr 1.0 to 1.4 mg/dL; unadjusted hazard ratio 2.54, 95% confidence interval 1.73 to 3.71, P<0.0001 for SCr >1.4 mg/dL). On multivariable analysis, we found a significant interaction between SCr and right atrial pressures (interaction P<0.0001); increased SCr best predicted death in patients with right atrial pressure <10 mm Hg. Renal dysfunction is associated with a worse hemodynamic profile and is an independent predictor of mortality in PAH. Measurement of SCr is practical and offers a simple way to noninvasively predict outcome.


[Effect of saffron aqueous extract on the level of blood glucose in experimental diabetes mice].
To investigate the effects of saffron aqueous extract (SE) on blood glucose, lipid and pancreatic tissue in streptozocin-induced diabetes mice. Diabetes mellitus mice were established by intraperitoneal injection of streptozocin (60 mg/kg) for two consecutive days. The 30 well-established diabetes mice were randomly divided into three groups(n=10):diabetic mellitus (DM) group, SE treated (SE) group and positive control (metformin hydrochloride, MH) group. Another ten normal mice were selected as normal control (NC) group. The mice in SE and MH groups were intragastrically administered with SE 100 mg/kg or MH 100 mg/kg once a day for 6 weeks, mice in DM and NC were given normal saline. The amount of food-intake, water consumption and body weight were measured weekly, the changes of the indicators including fasting blood glucose (FBG), oral glucose-tolerance test (OGTT), glycated serum protein (GSP), insulin (INS) and blood lipid were determined after 6 weeks of continuous administration. The pathologic changes in the pancreas tissues were detected by HE staining. Compared with the normal control group, the amount of food-intake, water consumption, area under the curve, FBG, GSP, and total cholesterol (TC) were significantly increased, while fasting weight, INS and high density lipoprotein cholesterol (HDL-c) were dramatically decreased in DM group. Compared with DM group, the water consumption, FBG, area under the curve and TC in SE group were starkly declined with a notable elevation of HDL-c and INS. In addition, the biopsy from DM mice showed the structure of pancreas islet was destroyed and reduced, and vascular proliferation with irregular shape. The damaged pancreas was obviously repaired by giving SE. The saffron aqueous extract had efficacy on blood glucose and blood lipids reduction, improvement on damaged pancreas in streptozocininduced diabetic mice, which suggested that saffron could be used for the treatment in diabetes.


Late talkers and later language outcomes: Predicting the different language trajectories.
The aim of the current study was to investigate the risk factors present at 2 years for children who showed language difficulties that persisted from 2 to 10 years and difficulties that emerged later, at 10 years. Participants (n = 783) were drawn from the Raine Study in Western Australia. Patterns of change from 2 to 10 years were identified based on child performance on the Language Development Survey and the Clinical Evaluation of Language Fundamentals, respectively. Logistic regression analyses were conducted to test whether parental, family and child characteristics present at 2 years predicted poorer language outcomes at age 10. Across the 8 year period, 5.6% of the children displayed consistently low language skills, 5.9% improved skills and 23.2% deteriorated skills. Compared to children with consistently typical skills, the deteriorated group was more likely to have mothers who smoked during pregnancy, fathers with incomplete secondary education, low family income, poor early literacy environment and be male. Children showing consistently low language skills were more likely to have mothers who smoked during pregnancy than late talkers whose early delays improved. Results provide evidence of some modifiable risk factors at 2 years which are associated with language outcome.


Self-rated health in rural Appalachia: health perceptions are incongruent with health status and health behaviors.
Appalachia is characterized by poor health behaviors, poor health status, and health disparities. Recent interventions have not demonstrated much success in improving health status or reducing health disparities in the Appalachian region. Since one's perception of personal health precedes his or her health behaviors, the purpose of this project was to evaluate the self-rated health of Appalachian adults in relation to objective health status and current health behaviors. Appalachian adults (n = 1,576) were surveyed regarding health behaviors - soda consumer (drink ≥ 355 ml/d), or non-consumer (drink < 355 ml/d), fast food consumer (eating fast food ≥ 3 times/wk) or healthy food consumer (eating fast food < 3 times/wk), smoking (smoker or non-smoker), exercise (exerciser > 30 min > 1 d/wk) and sedentary (exercise < 30 min 1 d/wk), blood pressure medication (yes, no), and self-rated health. Blood pressure was measured through auscultation and serum cholesterol measured via needle prick. Weight status was based on BMI: normal weight (NW ≥ 18.5 and < 25.0), overweight (OW ≥ 25.0 and < 30.0), and obese (OB ≥ 30.0). Jaccard Binary Similarity coefficients, odds ratios, chi-square, and prevalence ratios were calculated to evaluate the relationships among self-rated health, objective health status, and health behaviors. Significance was set at p < 0.05. Respondents reported being healthy, while being sedentary (65%), hypertensive (76%), overweight (73%), or hyperlipidemic (79%). Between 57% and 66% of the respondents who considered themselves healthy had at least two disease conditions or poor health behaviors. Jaccard Binary Similarity coefficients and odds ratios showed the probability of reporting being healthy when having a disease condition or poor health behavior was high. The association between self-rated health and poor health indicators in Appalachian adults is distorted. The public health challenge is to formulate messages and programs about health and health needs which take into account the current distortion about health in Appalachia and the cultural context in which this distortion was shaped.


Simple approach for fast real-time line scan microscopic imaging.
A simple fast line scan microscopic imaging approach based on a wavelength-space-time mapping technique has been proposed. With a lab-made subpicosecond pulse laser with 10 dB bandwidth of 12 nm, we experimentally demonstrate a free-space optical apparatus designed for fast line scan imaging of microscopic objects. This system has a spatial resolution of 22 μm, field-of-view of 2.5 mm, and line scan rate of 20.9 MHz. By imaging a modified unitraveling carrier photodetector, we demonstrate the application of semiconductor device inspection for speeding up quality control.


Serotonin transporters upregulate with chronic cocaine use.
Cocaine potently inhibits serotonin (5-HT) reuptake in cell bodies and at nerve terminals and 5-HT has been implicated as a modulator of dopaminergic neurotransmission. Chronic use of cocaine may lead to a "serotonin-deficit" form of 5-HT dysregulation. We have examined the status of the 5-HT transporter (SERT) using ligand binding and autoradiographic methods in subgroups of cocaine overdose deaths. Quantitative autoradiography of [125I]RTI-55 was used to map and measure the effect of chronic cocaine use on SERT densities in the striatum, substantia nigra, amygdala, and adjacent paralimbic cortical areas of cocaine overdose (CO) victims with and without preterminal evidence of excited delirium (ED). SERT densities were elevated in the nucleus accumbens and throughout the anterior and posterior sectors of striatum in CO victims compared with age-matched and drug-free control subjects. In contrast, SERT densities were increased significantly in the anterior striatum, but not the posterior sectors in ED victims. Significant elevations in SERT were measured in the orbitofrontal gyrus (Brodmann area 11), the anterior portion of the insular cortex and the cingulate gyrus (Brodmann area 24) in CO and ED victims. Saturation binding site analysis demonstrated an increase in the density of RTI-55 binding sites with no change in the affinity of the radioligand for the SERT. Chronic cocaine exposure upregulated SERT densities in the substantia nigra of the CO, but not ED victims. The lack of SERT upregulation in the substania nigra and posterior striatum suggests the possibility of a distinct phenotype for fatal ED victims that exhibited an acute onset of bizarre and violent behavior prior to death. Adaptive changes in the SERT densities may contribute to depressed mood and drug craving associated with acute cocaine abstinence.


Understanding long-term changes in species abundance using a niche-based approach.
One of the major challenges to understanding population changes in ecology for assessment purposes is the difficulty in evaluating the suitability of an area for a given species. Here we used a new simple approach able to faithfully predict through time the abundance of two key zooplanktonic species by focusing on the relationship between the species' environmental preferences and their observed abundances. The approach is applied to the marine copepods Calanus finmarchicus and C. helgolandicus as a case study characterising the multidecadal dynamics of the North Sea ecosystem. We removed all North Sea data from the Continuous Plankton Recorder (CPR) dataset and described for both species a simplified ecological niche using Sea Surface Temperature (SST) and CPR Phytoplankton Colour Index (PCI). We then modelled the dynamics of each species by associating the North Sea's environmental parameters to the species' ecological niches, thus creating a method to assess the suitability of this area. By using both C. finmarchicus and C. helgolandicus as indicators, the procedure reproduces the documented switches from cold to warm temperate states observed in the North Sea.


Recombination between viral DNA and the transgenic coat protein gene of African cassava mosaic geminivirus.
Nicotiana benthamiana was transformed with three different constructs (pCRA1, pCRA2 and pJC1) containing the coat protein coding sequence of African cassava mosaic virus (ACMV). Transformed plants were inoculated with a coat protein deletion mutant of ACMV that induces mild systemic symptoms in control plants. Several inoculated plants of transgenic lines CRA1/3, CRA1/4, CRA2/1 and CRA2/2 developed severe systemic symptoms typical of ACMV. DNA analysis revealed that, in these plants, recombination had occurred between the mutant viral DNA and the integrated construct DNA, resulting in the production of recombinant virus progeny with 'wild-type' characteristics. No reversion of mutant to 'wild-type' virus was observed in pJC1-transformed plants. Recombinant virus from several transgenic plants was analysed by PCR and parts of DNA A of virus progeny were cloned. Sequence analysis revealed that only a few nucleotides were changed from the published sequence.


Occurrence of Trichosirocalus horridus (Coleoptera: Curculionidae) on native Cirsium altissimum versus exotic C. vulgare in North American tallgrass prairie.
Postrelease studies can provide data with which to evaluate expectations based on prerelease tests of biological control insects. In 2004, we observed Trichosirocalus horridus Panzer (Coleoptera: Curculionidae), the rosette weevil introduced into North America against Eurasian thistles, feeding on native tall thistle, Cirsium altissimum L. Spreng., in tallgrass prairie. In this study, we examined the rosette weevil's use of tall thistle, compared with its use of the co-occurring exotic bull thistle, C. vulgare (Savi) Tenore. For both thistle species, we quantified weevil frequency, abundance, and seasonal variation in incidence, using both timed observations at two sites over two growing seasons (2004, 2005) and dissections of thistle flowering shoots from 13 sites (2005). Based on prerelease information, we expected the Eurasian thistle to be the quantitatively preferred host plant for this Eurasian weevil. Instead, we found that both the frequency of infestation and the mean number of adult rosette weevils per plant were at least as high, and sometimes higher, on the native thistle as on the exotic thistle. Furthermore, adult weevil phenology coincided on the two host species. This study provides new quantitative evidence of nontarget feeding by another weevil released for thistle biological control; and it raises important questions for further research. We conclude that continued new releases, as well as augmentation of existing populations, of T. horridus should wait until more research is done on the impact of the nontarget occurrence now reported for this biological control insect.


Effect of the uvrD mutation on excision repair.
A pair of related Escherichia coli K-12 strains, one of which contains the uvrD101 mutation, were constructed and compared for ability to perform various steps in the excision repair of deoxyribonucleic acid damage inflicted by ultraviolet radiation. The results of this study indicated: (i) ultraviolet sensitivity in the uvrD101 mutant was greater than that of wild type but less than that measured in an incision-deficient uvrA mutant; (ii) host cell reactivation paralleled the survival data; (iii) postirradiation deoxyribonucleic acid degradation was virtually identical in the two strains; (iv) incision, presumably at the sites of pyrimidine dimers, proceeded normally in the uvrD101 strain; (v) excision of pyrimidine dimers was markedly reduced in both rate and extent in the uvrD101 mutant; (vi) the amount of repair resynthesis was the same in both strains, and there was no evidence of abnormally long repair patches in the uvrD mutant; and (vii) rejoining of incision breaks was slow and incomplete in the uvrD strain. These data suggest that the ultraviolet sensitivity conferred by the uvrD mutation arises from inefficient removal of pyrimidine dimers or from failure to close incision breaks. The data are compatible with the notion that the uvrD+ gene produce affects the conformation of incised deoxyribonucleic acid molecules.


[Ligamentoplasty using the peroneus brevis in the treatment of chronic instabilities of the ankle. Long-term review].
Sixteen cases in which a ligamentoplasty using the peroneus brevis have been followed up for more than 8 years. In half of the cases dorsi-flexion of the ankle was diminished and the subtaloid joint was stiff. There was no anterior drawer sign of the talus. No case of progressive degenerative arthritis of the ankle joint was seen.


Conceptualizing and measuring historical trauma among American Indian people.
This article reports on the development of two measures relating to historical trauma among American Indian people: The Historical Loss Scale and The Historical Loss Associated Symptoms Scale. Measurement characteristics including frequencies, internal reliability, and confirmatory factor analyses were calculated based on 143 American Indian adult parents of children aged 10 through 12 years who are part of an ongoing longitudinal study of American Indian families in the upper Midwest. Results indicate both scales have high internal reliability. Frequencies indicate that the current generation of American Indian adults have frequent thoughts pertaining to historical losses and that they associate these losses with negative feelings. Two factors of the Historical Loss Associated Symptoms Scale indicate one anxiety/depression component and one anger/avoidance component. The results are discussed in terms of future research and theory pertaining to historical trauma among American Indian people.


Genetics of suicide attempts in individuals with and without mental disorders: a population-based genome-wide association study.
Family studies have shown an aggregation of suicidal behavior in families. Yet, molecular studies are needed to identify loci accounting for genetic heritability. We conducted a genome-wide association study and estimated single nucleotide polymorphisms (SNP) heritability for a suicide attempt. In a case-cohort study, national data on all individuals born in Denmark after 1981 and diagnosed with severe mental disorders prior to 2013 (n = 57,377) and individuals from the general population (n = 30,000) were obtained. After quality control, the sample consisted of 6024 cases with an incidence of suicide attempt and 44,240 controls with no record of a suicide attempt. Suggestive associations between SNPs, rs6880062 (p-value: 5.4 × 10-8) and rs6880461 (p-value: 9.5 × 10-8), and suicide attempt were identified when adjusting for socio-demographics. Adjusting for mental disorders, three significant associations, all on chromosome 20, were identified: rs4809706 (p-value: 2.8 × 10-8), rs4810824 (p-value: 3.5 × 10-8), and rs6019297 (p-value: 4.7 × 108). Sub-group analysis of cases with affective disorders revealed SNPs associated with suicide attempts when compared to the general population for gene PDE4B. All SNPs explained 4.6% [CI-95: 2.9-6.3%] of the variation in suicide attempt. Controlling for mental disorders reduced the heritability to 1.9% [CI-95: 0.3-3.5%]. Affective and autism spectrum disorders exhibited a SNP heritability of 5.6% [CI-95: 1.9-9.3%] and 9.6% [CI-95: 1.1-18.1%], respectively. Using the largest sample to date, we identified significant SNP associations with suicide attempts and support for a genetic transmission of suicide attempt, which might not solely be explained by mental disorders.


Cyclic Nucleotide Monophosphates and Their Cyclases in Plant Signaling.
The cyclic nucleotide monophosphates (cNMPs), and notably 3',5'-cyclic guanosine monophosphate (cGMP) and 3',5'-cyclic adenosine monophosphate (cAMP) are now accepted as key signaling molecules in many processes in plants including growth and differentiation, photosynthesis, and biotic and abiotic defense. At the single molecule level, we are now beginning to understand how cNMPs modify specific target molecules such as cyclic nucleotide-gated channels, while at the systems level, a recent study of the Arabidopsis cNMP interactome has identified novel target molecules with specific cNMP-binding domains. A major advance came with the discovery and characterization of a steadily increasing number of guanylate cyclases (GCs) and adenylate cyclases (ACs). Several of the GCs are receptor kinases and include the brassinosteroid receptor, the phytosulfokine receptor, the Pep receptor, the plant natriuretic peptide receptor as well as a nitric oxide sensor. We foresee that in the near future many more molecular mechanisms and biological roles of GCs and ACs and their catalytic products will be discovered and further establish cNMPs as a key component of plant responses to the environment.


Dental hygiene intervention to prevent nosocomial pneumonias.
Nosocomial and ventilator associated pneumonias that plague critically ill, elderly and long-term care residents could be reduced with effective oral hygiene practices facilitated collaboratively between nurses and dental hygienists. Nosocomial pneumonias, specifically aspiration pneumonias and ventilator-associated pneumonias in the elderly and infirm have become a major health care issue, The provision of oral care in hospital and hospital-like facilities presents challenges that can prevent patients from receiving optimal oral care One sequela can be aspiration pneumonia which ranks first in mortality and second in morbidity among all nosocomial infections. Since aspiration pneumonia is linked to the colonization of oral bacteria in dental plaque and biofilm, it is time to look for creative solutions to integrating the expertise of dental hygienists into health care teams in these institutional settings. A comprehensive review of the literature was conducted regarding the etiology and prevalence of health care related pneumonias. Evidence describing the challenges and barriers that the nurses, nursing staff, and dental hygienists face in the provision of oral care in hospitals and long-term care facilities is provided. Intercollaborative solutions to providing optimal oral care in hospitals and long-term care facilities are suggested. Dental hygienists have the expertise and practice experience to provide oral care in hospitals, long-term care and residential facilities. They can contribute to solving oral care challenges through intercollaboration with other health care team members. Yet, there are long-standing systemic barriers that must be addressed in order to provide this optimal care. Dental hygienists becoming better assimilated within the total health care team in hospital and residential facilities can positively impact the suffering, morbidity and mortality associated with aspiration pneumonias.


Neutral, Negative, or Negligible? Changes in Patient Perceptions of Disease Risk Following Receipt of a Negative Genomic Screening Result.
Most individuals who undergo genomic screening will receive negative results or results not sufficient to warrant a clinical response. Even though a majority of individuals receive negative results, little is known about how negative results may impact individuals' perception of disease risk. Changes in risk perception (specifically reductions in perceived risk) may affect both probands and their family members if inaccurate information is communicated to family members. We surveyed patients who received negative results as part of their participation in a genomic screening study and assessed their perceptions of disease risk following receipt of results. Participants had either hyperlipidemia or colon polyps (or both) and received their negative genomic screening results by mail. Of 1712 total individuals recruited, 1442 completed the survey (84.2% completion rate). Approximately one quarter of individuals believed their risk for heart disease to be lower and approximately one third of individuals believed their risk for colon cancer to be lower after receiving and evaluating their negative genomic screening result. 78% of those who believed their risk for one or both diseases had declined had already shared or intended to share their result with family members. Our study suggests patients may interpret a negative genomic screening result as implying a reduction in their overall disease risk.


Dog hepatocytes are key effector cells in the liver innate immune response to Leishmania infantum.
Hepatocytes constitute the majority of hepatic cells, and play a key role in controlling systemic innate immunity, via pattern-recognition receptors (PRRs) and by synthesizing complement and acute phase proteins. Leishmania infantum, a protozoan parasite that causes human and canine leishmaniasis, infects liver by establishing inside the Kupffer cells. The current study proposes the elucidation of the immune response generated by dog hepatocytes when exposed to L. infantum. Additionally, the impact of adding leishmanicidal compound, meglumine antimoniate (MgA), to parasite-exposed hepatocytes was also addressed. L. infantum presents a high tropism to hepatocytes, establishing strong membrane interactions. The possibility of L. infantum internalization by hepatocytes was raised, but not confirmed. Hepatocytes were able to recognize parasite presence, inducing PRRs [nucleotide oligomerization domain (NOD)1, NOD2 and Toll-like receptor (TLR)2] gene expression and generating a mix pro- and anti-inflammatory cytokine response. Reduction of cytochrome P 450s enzyme activity was also observed concomitant with the inflammatory response. Addition of MgA increased NOD2, TLR4 and interleukin 10 gene expression, indicating an immunomodulatory role for MgA. Hepatocytes seem to have a major role in coordinating liver's innate immune response against L. infantum infection, activating inflammatory mechanisms, but always balancing the inflammatory response in order to avoid cell damage.


[Revascularizing osteotrepanation in combined surgical treatment of chronic lower limb ischemia].
Short and long-term results of revascularizing osteotrepanations by Zusmanovich's method performed in 80 patients with lower limb critical ischemia were analyzed. The operation is effective in distal and diffuse forms of atherosclerosis and inflammatory diseases if it is impossible to perform reconstruction. Isolated revascularizing osteotrepanation is indicated for patients with recurrence of critical ischemia who earlier have undergone conventional surgical procedures. Duplex sonography is the main method for diagnosis and determination of indications for surgery which provides an objective criterion of the treatment failure - the index of tibial arteries resistance.


Hepatic glucose sensing: does flux matter?
In this issue of the JCI, Denechaud et al. report studies investigating the role of the liver X receptors (LXRs) LXRalpha and LXRbeta in carbohydrate sensing by the liver (see the related article beginning on page 956). The results of this study, which utilized LXRalpha/beta double-KO mice, strongly contradict a recent Nature report that proposed that LXRalpha/beta sense glucose independent of metabolic flux. The reported findings further support a key role for the carbohydrate-responsive element-binding protein (ChREBP) in the regulation of lipogenic genes by glucose and dietary carbohydrates.


Reinforcing properties of stereoisomers of the putative sigma agonists N-allylnormetazocine and cyclazocine in rhesus monkeys.
There are many similarities in the effects of phencyclidine (PCP) and opioids with sigma agonist activity such as N-allylnormetazocine (SKF-10,047) and cyclazocine. Yet PCP and sigma agonists differ in reinforcing properties. PCP is reliably self-administered in animal models and has abuse liability in humans, whereas (+/-)-N-allylnormetazocine and (+/-)-cyclazocine are not self-administered or abused. Inasmuch as other research has demonstrated differences in the selectivity of PCP-like effects of the stereoisomers of opioids, this study compared the reinforcing properties of PCP to the reinforcing properties of isomers of N-allylnormetazocine and cyclazocine. Rhesus monkeys were trained to lever-press on a fixed-ratio 10 schedule for i.v. cocaine (50 micrograms/kg/injection). During substitution tests, cocaine was replaced with various doses of test drugs or vehicle. The drugs tested were PCP, (+/-)-N-allylnormetazocine, (-)-N-allylnormetazocine, (+)-N-allylnormetazocine, (+/-)-cyclazocine, (-)-cyclazocine and (+)-cyclazocine. PCP and the (+)-isomers of both benzomorphans, at one or more doses, were self-administered at rates which exceeded rates maintained by vehicle and which equaled or exceeded cocaine-maintained base-line rates. Neither the racemates nor the (-)-isomers of N-allylnormetazocine or cyclazocine were self-administered by the monkeys at rates which were greater than vehicle control levels of responding. Reinforcing doses of the three compounds resulted in injections evenly distributed across the 60-min session in a pattern similar to cocaine self-administration, whereas the racemic mixtures and the (-)-isomers produced negatively accelerated patterns of injections which resembled typical saline patterns of extinction responding. These data provide further evidence that the (+)-isomers of sigma agonists are more selective for PCP-like effects than the (-)-isomers in monkeys.


Sequencing Grade Tandem Mass Spectrometry for Top-Down Proteomics Using Hybrid Electron Capture Dissociation Methods in a Benchtop Orbitrap Mass Spectrometer.
Compared to traditional collision induced dissociation methods, electron capture dissociation (ECD) provides more comprehensive characterization of large peptides and proteins as well as preserves labile post-translational modifications. However, ECD experiments are generally restricted to the high magnetic fields of FTICR-MS that enable the reaction of large polycations and electrons. Here, we demonstrate the use of an electromagnetostatic ECD cell to perform ECD and hybrid ECD methods utilizing 193 nm photons (ECuvPD) or collisional activation (EChcD) in a benchtop quadrupole-Orbitrap mass spectrometer. The electromagnetostatic ECD cell was designed to replace the transfer octapole between the quadrupole and C-trap. This implementation enabled facile installation of the ECD cell, and ions could be independently subjected to ECD, UVPD, HCD, or any combination. Initial benchmarking and characterization of fragmentation propensities for ECD, ECuvPD, and EChcD were performed using ubiquitin (8.6 kDa). ECD yielded extensive sequence coverage for low charge states of ubiquitin as well as for the larger protein carbonic anhydrase II (29 kDa), indicating pseudo-activated ion conditions. Additionally, relatively high numbers of d- and w-ions enable differentiation of isobaric isoleucine and leucine residues and suggest a distribution of electron energies yield hot-ECD type fragmentation. We report the most comprehensive characterization to date for model proteins up to 29 kDa and a monoclonal antibody at the subunit level. ECD, ECuvPD, and EChcD yielded 93, 95, and 91% sequence coverage, respectively, for carbonic anhydrase II (29 kDa), and targeted online analyses of monoclonal antibody subunits yielded 86% overall antibody sequence coverage.


Synthesis of oxime ethers via a formal reductive O-H bond insertion of oximes to α-keto esters.
This study describes an efficient approach to access oxime ethers via P(III)-mediated O-H bond insertion reaction of oximes with α-keto esters. The strategy involves the protonation of in situ generated Kukhtin-Ramirez adducts, followed by SN2-type reaction. Important features include a good functional group tolerance, operational simplicity, and application to gram scale synthesis and the synthesis of an acaricide.


Morbidity from diarrhoea, cough and fever among young children in Nigeria.
Diarrhoea, cough and fever are the leading causes of childhood morbidity and mortality in sub-Saharan Africa. Despite it being a determinant of mortality in many developing countries, geographical location has seldom been considered as an explanatory factor for the large regional variations seen in the childhood morbidity attributed to these causes in this area. The relevant data collected in two Nigerian Demographic and Health Surveys, one in 1999 and the other in 2003, have now therefore been analysed and compared. The aim was to reveal and explore inequalities in the health of Nigerian children by mapping the spatial distribution of childhood morbidity associated with recent diarrhoea, cough and fever and accounting for important risk factors, using a Bayesian geo-additive model based on Markov-chain-Monte-Carlo techniques. Although the overall prevalences of recent diarrhoea, cough and fever recorded in 1999 (among children aged <3 years) were similar to those seen in 2003 (among children aged <5 years), the mapping of residual spatial effects indicated that, in each survey, the morbidity attributable to each of these causes varied, differently, at state level. Place of birth (hospital v. other), type of feeding (breastfed only v. other), parental education, maternal visits to antenatal clinics, household economic status, marital status of mother and place of residence were each significantly associated with the childhood morbidity investigated. In both surveys, children from urban areas were found to have a significantly lower risk of fever than their rural counterparts. Most other factors affecting diarrhoea, cough and fever differed in the two surveys. The risk of developing each of these three conditions increased in the first 6-8 months after birth but then gradually declined. The analysis explained a significant share of the pronounced residual spatial effects. Maps showing the prevalences of diarrhoea, cough and fever in young children across Nigeria were generated during this study. Such maps should facilitate the development of policies to fulfil the Millennium Development Goals in Nigeria and throughout sub-Saharan Africa.


Findings from a prospective cohort study evaluating the effects of International Health Advisors' work on recently settled migrants' health.
Several interventions have been carried out to tackle health inequalities between migrant groups, especially refugees, and native-born European populations. These initiatives are often address language or cultural barriers. One of them is the International Health Advisors (IHA) in Sweden; a peer education intervention aimed at providing health information for recently settled migrants. It is known that social determinants, such as educational level and access to social capital, affect health. Social determinants may also affect how health information is received and transformed into practice. The aims of this study was to a) assess the impact of the IHA on recently settled migrants' self-reported health status, and received health information; b) determine the moderating role of educational level and social capital; and c) critically discuss the outcomes and suggest implications for health promotion practice. The study was designed as a prospective cohort study. A postal questionnaire translated to Arabic was sent to recently settled Iraqi migrants in eight counties in Sweden, in May 2008 and May 2010. Two of the counties were exposed to the intervention, and six were used as references. The proportion of individuals who reported that they had received information on healthy diet and physical exercise was higher in the intervention group than in the non-intervention group (OR 2.31, 95% CI 1.02-5.22), after adjustments. Low social participation was negatively associated with deteriorated or unchanged health needs (OR 0.47, 95% CI 0.24-0.92). No other statistically significant differences in health outcomes could be observed between the groups. No signs of effect modification on this association by social capital or educational level could be found. Health information provided by the IHA increased self-reported level of knowledge on healthy diet and physical exercise. The interpretation of the observed negative association between low social participation and deteriorated or unchanged health needs is that participation was limited to one's own social group, and therefore had limited positive influence on health seeking behaviour. The lack of measurable improvements in health status could be explained by limitations in the study, in the theoretical assumptions underlying the intervention, and in the implementation of the intervention. Further research is needed to understand success factors in health promoting interventions among recently settled migrants better.


Control of antiviral defenses through hepatitis C virus disruption of retinoic acid-inducible gene-I signaling.
Hepatitis C virus (HCV) is a major human pathogen that infects 170 million people. A hallmark of HCV is its ability to establish persistent infections reflecting the evasion of host immunity and interference with alpha/beta-IFN innate immune defenses. We demonstrate that disruption of retinoic acid-inducible gene I (RIG-I) signaling by the viral NS3/4A protease contributes to the ability of HCV to control innate antiviral defenses. RIG-I was essential for virus or HCV RNA-induced signaling to the IFN-beta promoter in human hepatoma cells. This signaling was disrupted by the protease activity of NS3/4A, which ablates RIG-I signaling of downstream IFN regulatory factor 3 and NF-kappaB activation, attenuating expression of host antiviral defense genes and interrupting an IFN amplification loop that otherwise suppresses HCV replication. Treatment of cells with an active site inhibitor of the NS3/4A protease relieved this suppression and restored intracellular antiviral defenses. Thus, NS3/4A control of RIG-I supports HCV persistence by preventing IFN regulatory factor 3 and NF-kappaB activation. Our results demonstrate that these processes are amenable to restoration through pharmacologic inhibition of viral protease function.