Social Distress among Cancer Patients: Differential Effects of Risk Factors and Attenuating Role of Culturally Specific Social Support.
Introduction: We investigated the association between social distress or toxicity and patients' clinical conditions, demographic characteristics, and social support and networks, and whether this association differs along the distribution of patients' distress levels. This study included 156 patients treated at King Abdulaziz Medical City, Riyadh, Saudi Arabia. Methods: We used the previously validated Social Toxicity Assessment Tool in Cancer (STAT-C) to assess cancer patients' distress. We analyzed distress level, the outcome variable of interest, and covariates to show distribution and identify associations. We then used logistic quantile regression for bounded outcomes to assess the association between social distress or toxicity and patients' clinical conditions, demographic characteristics, and social support and network. As an extension, we examined the interaction between disease status and social support, focusing on the moderating role of social support in attenuating the impact of disease status on social distress. Results: The median age of the patients was 51.2 (SD = 21.4, range 22 to 89), with 48.1% being older than 50 years. Of the 156 cancer patients analyzed, 82 (52.6%) were classified as burdened, and 50% of those with uncontrolled disease status were socially distressed. However, there were more socially distressed patients diagnosed within a year and patients undergoing treatment. There was a greater number of patients who shared their diagnosis with family, colleagues, and neighbors with social distress. The odds of suffering from social distress were higher in younger patients (50 years or younger) than in older patients. Social distress was lower in patients who underwent combined chemotherapy, surgery, and radiation compared with patients who received a single treatment regimen (OR = 0.65, CI, -0.820 to -0.036, p = 0.033). The odds of social distress were 67% higher in patients diagnosed within one year than in patients diagnosed more than one year prior (OR = 1.664, CI, 0.100-0.918, p = 0.015). Patients with uncontrolled disease conditions who shared their diagnosis and treatment with social networks were 48% less likely to experience social distress. Thus, sharing cancer diagnoses with social networks has a statistically significant moderating effect by attenuating the impact of disease status on social distress. Conclusion: Understanding the risk factors for social distress may be important for cancer management. Additionally, identifying the moderating role that patients' sharing of cancer diagnoses in social networks plays in attenuating the impact of disease status on social distress may provide healthcare providers with valuable insights for holistic culture-specific care.


Hoxa9 transduction induces hematopoietic stem and progenitor cell activity through direct down-regulation of geminin protein.
Hoxb4, a 3'-located Hox gene, enhances hematopoietic stem cell (HSC) activity, while a subset of 5'-located Hox genes is involved in hematopoiesis and leukemogenesis, and some of them are common translocation partners for Nucleoporin 98 (Nup98) in patients with leukemia. Although these Hox gene derivatives are believed to act as transcription regulators, the molecular involvement of the Hox gene derivatives in hematopoiesis and leukemogenesis remains largely elusive. Since we previously showed that Hoxb4 forms a complex with a Roc1-Ddb1-Cul4a ubiquitin ligase core component and functions as an E3 ubiquitin ligase activator for Geminin, we here examined the E3 ubiquitin ligase activities of the 5'-located Hox genes, Hoxa9 and Hoxc13, and Nup98-Hoxa9. Hoxa9 formed a similar complex with the Roc1-Ddb1-Cul4a component to induce ubiquitination of Geminin, but the others did not. Retroviral transduction-mediated overexpression or siRNA-mediated knock-down of Hoxa9 respectively down-regulated or up-regulated Geminin in hematopoietic cells. And Hoxa9 transduction-induced repopulating and clonogenic activities were suppressed by Geminin supertransduction. These findings suggest that Hoxa9 and Hoxb4 differ from Hoxc13 and Nup98-Hoxa9 in their molecular role in hematopoiesis, and that Hoxa9 induces the activity of HSCs and hematopoietic progenitors at least in part through direct down-regulation of Geminin.


Escape of Mexican Fruit Flies from Traps Baited with CeraTrap and Effect of Lure Feeding on Reproduction and Survival.
McPhail type traps are commonly used to catch tephritid pests using liquid baits as attractants. Unfortunately, these traps allow that some flies escape after feeding on the bait. Recently, a long-lasting and nontoxic enzymatic-hydrolyzed protein, known commercially as CeraTrap, has been introduced as a novel liquid bait. Because feeding on hydrolyzed proteins can improve reproduction in many tephritids, questions remain as to whether CeraTrap could provide a nutritious meal to flies that ingest the bait but escape the trap. In this study, we analyzed the efficacy of CeraTrap baited traps in capturing Anastrepha ludens flies of two different physiological states and two different diets. We also explored the effect of feeding on CeraTrap on the mating performance of males, ovarian development and fecundity of females and fly survival. Results demonstrated that CeraTrap attracted more protein-deprived and immature flies of both sexes and 2-30% of flies that entered the trap managed to escape. However, after feeding on the bait for 24 h, males exhibited lower mating success, longer latency to mate and longer copula durations than males fed on either sugar or sugar+ protein. Interestingly, females fed CeraTrap for 24 h developed ovaries as large as those females fed on sugar+ protein, however, they laid no eggs. When feeding continuously on CeraTrap, flies exhibited shorter lifespans and females laid no eggs. These findings indicate that, although some flies can ingest the bait and escape from the trap, their reproductive traits are negatively affected.


Working longer.
New NHS pension scheme rules come into force in April, and one of the changes is a higher retirement age. To help staff plan for this, NHS Employers' Working Longer Group will publish a suite a resources to help employees understand and prepare for the implications of working into their late sixties. In the meantime, a factsheet with information for staff can be found at tinyurl.com/mjb4u5f and for employers at http://www.nhsemployers.org/your-workforce/need-to-know/working-longer-group.


Determinants of anti-fibrotic response to mineralocorticoid receptor antagonist therapy: insights from the Eplerenone Post-Acute Myocardial Infarction Heart Failure Efficacy and Survival Study (EPHESUS) and Early Eplerenone Treatment in Patients with Acute ST-elevation Myocardial Infarction without Heart Failure (REMINDER) trials.
After myocardial infarction complicated by heart failure or diabetes, eplerenone (compared to placebo) significantly decreases amino-terminal propeptide of type III procollagen (PIIINP). Determining the subset of patients who are more prone to have a decrease in PIIINP and those who may respond better to the anti-fibrotic effects of mineralocorticoid receptor antagonists (MRA) therapy may be relevant for a personalized treatment approach. The aim of this study is to identify predictors of a PIIINP decrease and assess potential subgroups of "responders" to eplerenone. Clinical factors and biomarkers were evaluated as predictors of a PIIINP decrease from randomization to month 9 in 323 patients from the biomarker substudy of Eplerenone Post-Acute Myocardial Infarction Heart Failure Efficacy and Survival Study (EPHESUS). Additionally, the association between PIIINP decrease and the composite of cardiovascular (CV) death or CV hospitalization were also explored. External validation was performed in the REMINDER trial. Female sex, eplerenone, reperfusion therapy, potassium < 4 mmol/L, circulating levels of PIIINP ≥ 3.6 ng/mL and PINP ≥ 27 ng/mL predicted a PIIINP decrease (AUC = 0.75). Randomization PIIINP showed a significant interaction with the treatment allocation: patients with PIIINP ≥ 3.6 ng/mL had a better response (decrease in PIIINP) to eplerenone (OR for PIIINP ≥ 3.6 = 2.9, 95% CI 1.46-5.89, p = 0.003) and OR for PIIINP < 3.6 = 1.09, 95% CI 0.55-2.2, p = 0.8; interactionp = 0.026). These findings were internally robust using another statistical approach (LOESS). External validation showed good discrimination (AUC = 0.70). There was a tendency toward a lower rate of CV death/CV hospitalizations in patients with decreased PIIINP (adjusted HR = 0.52, 95% CI 0.26-1.02, p = 0.058). In patients who had a myocardial infarction, clinical factors used in combination and treatment with eplerenone were associated with a PIIINP decrease. Interestingly, higher randomization PIIINP levels might help in identifying patients more prone to have an "anti-fibrotic response" when treated with MRAs. Predictors of an antifibrotic response after MI complicated by HF. Several clinical factors and biomarkers predicted a PIIINP decrease after an MI complicated by HF. There was a significant interaction between baseline PIIINP levels and eplerenone treatment: patients with baseline PIIINP ≥ 3.6 mmol/L treated with eplerenone had the best response (PIIINP decrease).


Extremely superhydrophobic surfaces with micro- and nanostructures fabricated by copper catalytic etching.
We demonstrate a simple method for the fabrication of rough silicon surfaces with micro- and nanostructures, which exhibited superhydrophobic behaviors. Hierarchically rough silicon surfaces were prepared by copper (Cu)-assisted chemical etching process where Cu nanoparticles having particle size of 10-30 nm were deposited on silicon surface, depending on the period of time of electroless Cu plating. Surface roughness was controlled by both the size of Cu nanoparticles and etching conditions. As-synthesized rough silicon surfaces showed water contact angles ranging from 93° to 149°. Moreover, the hierarchically rough silicon surfaces were chemically modified by spin-coating of a thin layer of Teflon precursor with low surface energy. And thus it exhibited nonsticky and enhanced hydrophobic properties with extremely high contact angle of nearly 180°.


Evaluation of a clinical skills orientation program for residents.
Our purpose was to implement and evaluate an orientation program for residents, focusing on outpatient clinical skills. Eleven of 12 residents participated in a clinical skills orientation program immediately preceding the academic year. The skill stations included evaluation of abnormal uterine bleeding, intrauterine device insertion, basic infertility evaluation, endometrial and vulvar biopsies, pelvic organ prolapse quantification examination, hysterosalpingography and office hysteroscopy, ultrasound scanning, labor and delivery triage, and clinic administrative responsibilities. Before test, after test, and anonymous resident evaluations were used to evaluate the program. First-year residents demonstrated a statistically significant increase in posttest scores compared to pretest scores (42.5% vs 71.3%, P=.003). Only first-year resident posttest scores for the labor and delivery triage and basic infertility evaluation stations demonstrated statistically significant increases over pretest scores (14.3% vs 46.4%, P=.009; and 41.7% vs 83.3%, P=.049, respectively). Sixty-four percent of the residents rated the program as "very helpful." Most residents felt that the program was well organized and that the facilities were conducive to learning; all of the participants recommended an annual clinical orientation program. A clinical skills orientation program was well received and strongly desired by residents. First-year residents appeared to benefit the most from this orientation.


Gain in body fat is inversely related to the nocturnal rise in serum leptin level in young females.
Both genetic and environmental factors contribute to adolescent obesity. Evidence of a genetic basis for obesity development is substantial, although the exact mechanism of action has yet to be identified. The purpose of this study was to document the circadian rhythmicity of the serum leptin level in young females and to assess the impact of the change in body fat stores during growth on the nocturnal rise in the serum leptin level with implications for obesity traits. There was a significant rise in serum leptin at midnight and 0400 h, suggesting a diurnal variation in serum leptin concentrations (ANOVA F ratio = 6.2; P < 0.0001). There was also a strong association between relative total body fat and the average daytime serum leptin level (r = 0.78; P < 0.0001). The percent increase in the nocturnal leptin concentration was inversely related to the percent gain in total body fat (r = 0.45; P < 0.024). Forward stepwise regression analysis selected the change in total body fat over a 6-month interval as the most powerful determinant of the percent increase in the nocturnal leptin concentration (partial R2 = 0.203; beta = -0.450; SE of beta = 0.186; t = -2.418; P < 0.024). If the lack of a nocturnal rise in serum leptin persists over a longer period of time, it may have implications for the development of obesity, presumably by inadequate suppression of nighttime appetite.


HECT Domain and RCC1-Like Domain-Containing Protein 5 (HERC-5) Gene Polymorphisms in HIV-1-Infected Individuals: A Study from India.
HECT domain and RCC1-like domain-containing protein 5 (HERC-5) is one of the novel host restriction factors that is known to inhibit HIV release in vitro. Polymorphisms in other host restriction factors have been associated with HIV infection and disease progression. However, no report is available on the HERC-5 polymorphism in HIV-infected individuals. We studied the HERC-5 gene polymorphism in HIV-infected individuals and explored whether it is associated with different disease outcomes. Genomic DNA was isolated from 41 HIV-1 progressors, 39 long-term nonprogressors, and 74 HIV seronegative healthy donors for amplification of HERC5 Exon-18 and other regulatory regions followed by sequencing. We found no genetic variation in the known single-nucleotide polymorphism (SNP)-rs34457268 (Exon-18) of HERC-5 in HIV-infected individuals. Instead, a synonymous mutation at rs6857425 (T-C) was present in the same region among all study groups (p > .05), irrespective of their HIV status. We further noted two novel SNPs in Intron-18 region. To the best of our knowledge, this is first study to report the HERC5 gene polymorphism among HIV-infected groups.


The effect of prepartum feeding and lying space on metabolic health and immune function.
The determinants of metabolic and reproductive health disorders in the peripartum period and the degree to which feeding and lying space and management can influence health are only partially understood. The objective of this randomized controlled study was to determine whether providing noncompetitive feeding and lying access in the close-up dry period improves health and immune function. Forty-eight Holstein cows of all parities were randomly assigned to a treatment group of 6 to 10 cows in 1 pen with either 80% cows to stalls and 90 cm of feeding space per cow (understocked) or 120% stocking density and 45 cm of feeding space per cow (overstocked) for 3 wk before expected calving. All cows wore electronic data loggers to monitor daily standing and lying time. Video recordings representing d 7 to 9 after group formation were reviewed, and a competition index (C_Ind) was calculated for each cow by dividing the number of times a cow displaced another as an actor by its total number of actor and reactor displacements. Cows were categorized as high success (C_Ind ≥0.6), moderate success (0.4 ≤ C_Ind <0.6), or low success (C_Ind <0.4). Weekly blood samples measured nonesterified fatty acids, β-hydroxybutyrate, calcium, glucose, albumin, aspartate aminotransferase, bilirubin, haptoglobin, insulin, and insulin-like growth factor-1 from 3 wk before to 5 wk after calving. Measures of innate immune function (neutrophil phagocytosis and oxidative burst) were assessed at -2, -1, 1, 2, 3, and 5 wk relative to calving. Liver biopsies were collected at wk 1 and 3. Cows in the understocked group spent significantly more time per day lying; the back-transformed least squares means and 95% confidence interval were 14.8 h (13.9-15.6) versus 12.8 h (12.0-13.7). Controlling for parity, there was no difference between treatments in β-hydroxybutyrate, nonesterified fatty acids, glucose, insulin, insulin-like growth factor 1, aspartate aminotransferase, bilirubin, or haptoglobin concentrations. Throughout the study, cows in the understocked treatment had higher mean calcium and tended to have higher albumin and at 3 wk after calving tended to have lower mean liver triacylglycerol content. Overall, there was no treatment effect on phagocytosis, but cows with a higher C_Ind in the understocked treatment group had greater oxidative burst function. There was no effect of treatment on endometritis. Despite increased competition and lower lying time, the expected harmful effects of crowding and competition on metabolic indicators and innate immune function were mostly not observed. Although this does not refute the importance of access to feeding and lying space, these results indicate that metabolic and reproductive health is more complex than can be explained solely by exposure to what are understood to be best practices for space allowances.


Cerebellins are differentially expressed in selective subsets of neurons throughout the brain.
Cerebellins are secreted hexameric proteins that form tripartite complexes with the presynaptic cell-adhesion molecules neurexins or 'deleted-in-colorectal-cancer', and the postsynaptic glutamate-receptor-related proteins GluD1 and GluD2. These tripartite complexes are thought to regulate synapses. However, cerebellins are expressed in multiple isoforms whose relative distributions and overall functions are not understood. Three of the four cerebellins, Cbln1, Cbln2, and Cbln4, autonomously assemble into homohexamers, whereas the Cbln3 requires Cbln1 for assembly and secretion. Here, we show that Cbln1, Cbln2, and Cbln4 are abundantly expressed in nearly all brain regions, but exhibit strikingly different expression patterns and developmental dynamics. Using newly generated knockin reporter mice for Cbln2 and Cbln4, we find that Cbln2 and Cbln4 are not universally expressed in all neurons, but only in specific subsets of neurons. For example, Cbln2 and Cbln4 are broadly expressed in largely non-overlapping subpopulations of excitatory cortical neurons, but only sparse expression was observed in excitatory hippocampal neurons of the CA1- or CA3-region. Similarly, Cbln2 and Cbln4 are selectively expressed, respectively, in inhibitory interneurons and excitatory mitral projection neurons of the main olfactory bulb; here, these two classes of neurons form dendrodendritic reciprocal synapses with each other. A few brain regions, such as the nucleus of the lateral olfactory tract, exhibit astoundingly high Cbln2 expression levels. Viewed together, our data show that cerebellins are abundantly expressed in relatively small subsets of neurons, suggesting specific roles restricted to subsets of synapses.


[Pain Signal Processing in the Spinal Dorsal Horn and Glial Cells].
Pain signals from the skin are transmitted to the spinal dorsal horn via primary afferent sensory neurons, appropriately processed, and transmitted to the brain. Neuropathic pain is a pathological condition that results from structural and functional changes that occur after nerve damage. Glial cells are crucial to these changes and play important roles in developing and maintaining neuropathic pain. This review article will outline glial cells' mechanism of action in neuropathic pain and discuss the development of potential analgesics.


PDD symptoms in ADHD, an independent familial trait?
The aims of this study were to investigate whether subtle PDD symptoms in the context of ADHD are transmitted in families independent of ADHD, and whether PDD symptom familiality is influenced by gender and age. The sample consisted of 256 sibling pairs with at least one child with ADHD and 147 healthy controls, aged 5-19 years. Children who fulfilled criteria for autistic disorder were excluded. The Children's Social Behavior Questionnaire (CSBQ) was used to assess PDD symptoms. Probands, siblings, and controls were compared using analyses of variance. Sibling correlations were calculated for CSBQ scores after controlling for IQ, ADHD, and comorbid anxiety. In addition, we calculated cross-sibling cross-trait correlations. Both children with ADHD and their siblings had higher PDD levels than healthy controls. The sibling correlation was 0.28 for the CSBQ total scale, with the CSBQ stereotyped behavior subscale showing the strongest sibling correlation (r = 0.35). Sibling correlations remained similar in strength after controlling for IQ and ADHD, and were not confounded by comorbid anxiety. Sibling correlations were higher in female than in male probands. The social subscale showed stronger sibling correlations in elder than in younger sibling pairs. Cross-sibling cross-trait correlations for PDD and ADHD were weak and not-significant. The results confirm that children with ADHD have high levels of PDD symptoms, and further suggest that the familiality of subtle PDD symptoms in the context of ADHD is largely independent from ADHD familiality.


Novel MLH1 frameshift mutation in an extended hereditary nonpolyposis colorectal cancer family.
To present novel frameshift mutation c.31delC [p.L11X] in the MLH1 gene identified in an extended Bulgarian hereditary non-polyposis colorectal cancer (HNPCC) family and to analyze the molecular and clinical findings within the pedigree concerning the proposal of adequate individual prophylactic strategy for all mutation carriers. The pedigree of the family consists of 42 members in four generations. Search for mutations in the MLH1 and hMSH2 genes was performed in the proband. After PCR amplification of all exons including flanking intronic regions, amplicons were directly sequenced. The mutation was found in nine from the thirteen pedigree members who signed informed consent to participate in the study. In three adenocarcinomas, microsatellite instability and lack of the MLH1 protein expression were detected. The only one tubulovillous adenoma analyzed was microsatellite stable and the MLH1 protein showed an intact staining. The newly described mutation c.31delC is HNPCC causative. Besides the typical clinical features of the syndrome, we found a specific pathologic manifestation such as moderate to high differentiated adenocarcinomas of the colon. One of the mutation carriers developed a benign giant cell soft tissue tumor. The primary tumor localizations were frequently extracolonic and detailed yearly gastrointestinal and gynecological examinations have been proposed to the mutation carriers. We emphasize the importance of including the HNPCC genetic counseling and testing as well in the following surveillance of all patients at risk in the services covered by the health insurance in Bulgaria.


Calcium mobilisation and CCK secretion induced by modified fatty acids and latex microspheres reveal dual receptor mechanisms for lipid stimulation of STC-1 cells.
How fatty acids stimulate enteroendocrine cells to release cholecystokinin (CCK) is largely unknown. Recently, we proposed that the murine enteroendocrine cell line, STC-1, responds to insoluble fatty acid aggregates rather than fatty acid monomers in solution. This hypothesis led to two testable predictions. First, other insoluble particles of similar size but unrelated to fatty acid may be able to stimulate STC-1 cells in a similar fashion to dodecanoic acid and second, fatty acid sensing in STC-1 cells should be fairly insensitive to chemical modifications of the fatty acid as long as these modifications do not greatly alter the ability of the molecule to form insoluble aggregates. We used several analogues of dodecanoic acid and several varieties of latex microsphere (varying in size and surface charge) to see whether the predictions of our model hold. We found that while there was at least one latex microsphere that could induce CCK secretion and calcium mobilisation in STC-1 cells, there was a very poor correlation between the presence of insoluble aggregates and a cellular response. Instead the most important property, determining the potency of fatty acid analogues as stimulants of CCK secretion, was their amphipathicity. Removal of either the polar head or lipophilic tail completely abolished the ability of a given fatty acid analogue to stimulate STC-1 cells. These data suggested that while fatty acids can stimulate cells as aggregates, they may also be acting in monomeric form with the oil:water partitioning coefficient playing a crucial role. We finally resolved this issue with the observation that the sulfate ion greatly altered the response of STC-1 cells to monomeric dodecanoic acid. In the presence of sulfate, STC-1 cells will only respond to dodecanoic acid aggregates whereas when sulfate is replaced with chloride the cells clearly respond to dodecanoic acid monomers which are completely in solution. In summary, we propose that dodecanoic acid can stimulate STC-1 cells via two separate pathways one involving fatty acid monomers in solution and one involving fatty acid aggregates. Which pathway dominates depends on the presence of sulfate in the extracellular medium.


The fine structure of the proximal growth plate and metaphysis of the avian tibia: endochondral osteogenesis.
The ultrastructure of endochondral osteogenesis from the proximal end of tibias of 7 weeks old normal White Leghorn and broiler chickens is described. Little, if any, ultrastructural differences exists between these two strains of birds. In the growth plate, proliferation of chondrocytes and their matriceal production is approximately matched by resorption. Erosion of the metaphyseal aspect of the growth plate is accomplished under two sets of circumstances: firstly, when mineralization of the cartilaginous matrix is so scanty as to be discernible only by electron microscopy, and secondly, when calcification is gross and easily appreciated by light microscopy. The former process appears to be accomplished in the main by perivascular mononuclear cells and the latter, to a great extent, by chondroclasts. It appears that metaphyseal blood vessels expand, by saccular protrusions, into pre-existing spaces or those created by cellular erosion. Cones or plates of calcified cartilage, which extend into the metaphysis, act as a template for initial deposition of bone. Osteoblasts recently surrounded by osteoid have adjacent to their plasmalemma a pericellular sheath which is composed of ill-defined clumps of amorphous material as well as some incompletely aggregated collagenous fibrils. Beyond this sheath collagenous fibrils and fibres, with distinct periodicities (ranging from 57 to 69 nm) are observed. Towards the diaphysis the range of the periodicity of fibres in osteoid narrows to 64-69 nm.


Accelerated Antidepressant Response in Geriatric Inpatients.
Six elderly depressed inpatients were treated for 2 weeks with low doses of nortriptyline and fluoxeline in combination. All patients showed significant reductions in depressive symptoms with minimal side effects. Clinical improvement appeared unrelated to whether plasma levels of nortriptyline were therapeutic. The implications of this regimen for accelerated antidepressant response as well as the limitations of this report are discussed.


Is it all in the act?
Struggling to breathe is one of the most distressing symptoms to have, or to witness. Treating acute shortness of breath is rewarding for both the patient and the doctor. The patient often takes at least the initial dose of medication under your (or your practice nurse's) supervision. You know it was taken and taken correctly.


5S ribosomal RNA genes in six species of Mediterranean grey mullets: genomic organization and phylogenetic inference.
This paper describes a study of the 5S ribosomal RNA genes (5S rDNA) in a group of 6 species belonging to 4 genera of Mugilidae. In these 6 species, the relatively short 5S rDNA repeat units, generated by PCR and ranging in size from 219 to 257 bp, show a high level of intragenomic homogeneity of both coding and spacer regions (NTS-I). Phylogenetic reconstructions based on this data set highlight the greater phylogenetic and genetic diversity of Mugil cephalus and Oedalechilus labeo compared with the genera Liza and Chelon. Comparative sequence analysis revealed significant conservation of the short 5S rDNA repeat units across Chelon and Liza. Moreover, a second size class of 5S rDNA repeat units, ranging from roughly 800 to 1100 bp, was produced in the Liza and Chelon samples. Only short 5S rDNA repeat units were found in M. cephalus and O. labeo. The sequences of the long 5S rDNA repeat units, obtained in Chelon labrosus and Liza ramada, differ owing to the presence of 2 large insertion/deletions (indels) in the spacers (NTS-II) and show considerable sequence identity with NTS-I spacers. Interspecific sequence variation of NTS-II spacers, excluding the indels, is low. Southern-blot hybridization patterns suggest an intermixed arrangement of short and long repeat units within a single chromosome locus.


Involvement of endogenous noradrenaline release in methylene blue-induced contraction of isolated rabbit aorta.
The vasocontractile response to methylene blue (Meb) was investigated in isolated rabbit aorta. Meb (1-100 microM) induced a slowly developing contraction after a long latency in rabbit aortic strip. The maximal contraction was obtained by 50 microM Meb, which corresponded to 1 microM noradrenaline (NA)-induced contraction. Once the maximal contraction was induced by Meb, the strip completely lost the contractile response to a further application of Meb. The usual NA-induced contraction, however, could be observed in such a Meb-insensitive aortic strip. Meb-induced contractions were not affected by atropine, diphenhydramine, methysergide, indomethacin, nordihydroguaiaretic acid and removal of endothelial cells from the aortic strip, but they were abolished by prazosin. In aortic strips from rabbits pretreated with reserpine (3.0 mg/kg, i.m.) for a day, Meb failed to induce contraction. Meb evoked the [3H] release from [3H]NA-preloaded aortic strips. In high performance liquid chromatographic analysis, a considerable amount of NA was found in the bathing fluid of the aortic strip in the presence of Meb. In addition, Meb pretreatment inhibited [3H]NA uptake by the aortic strip and abolished the contractile response to an electrical stimulation of adrenergic nerve terminals. Although Meb decreased the basal level of cyclic GMP in the aortic strip, Meb-induced [3H]NA release from the aortic strip was not affected by 8-bromo cyclic GMP. These results suggest that Meb-induced contraction of rabbit aorta is due to the release of endogenous NA from its storage pools of intramural adrenergic nerves through an independent mechanism of its cyclic GMP lowering effect. In addition, incubation of aortic strips with Meb resulted in depleting the storage NA and blocking the nerve function, suggesting that Meb might be useful for a pharmacological tool as an adrenergic neuron blocking agent in vitro.


Association between atmospheric ozone levels and damage to human nasal mucosa in Florence, Italy.
We evaluated the effects of urban air pollutants on human nasal mucosa over an 8-month period on 102 subjects living in Florence, Tuscany, Italy. A group of subjects living in a city with a lower level of pollution (Sassari, Sardinia, Italy) was also analyzed. Nasal mucosa cells were harvested by brushing, a noninvasive procedure. Half of the cells were used for genotoxicity studies using the alkaline comet assay, and half for morphological studies. The levels of DNA damage in the nasal mucosa were considerably higher (+73%) in the subjects living in Florence than in Sassari. High levels of atmospheric ozone in Florence air correlated with DNA damage, and to the prevalence of inflammatory pathologies of the upper respiratory tract, although the ozone concentrations were below the Italian recommended attention level. Furthermore, higher levels of DNA damage were correlated with a dysfunction in the ability to maintain a normal epithelial cell structure. These data suggest an association between ozone air levels and damage in the upper respiratory tract. It remains unclear whether ozone itself or other associated pollutants are responsible for the observed alterations.


Association of the bitter taste receptor gene TAS2R38 (polymorphism RS713598) with sensory responsiveness, food preferences, biochemical parameters and body-composition markers. A cross-sectional study in Italy.
This study examined the relationship between TAS2R38 gene polymorphism (RS713598), G/G, C/G or C/C genotype, and sensory responsiveness, food preferences, biochemical parameters and body composition in a cross-sectional study in 118 adults (24 men and 94 women). The frequencies of C/C, G/G and C/G were respectively 20.3%, 29.7% and 50.0%. As regards taste responsiveness, subjects with G-allele had a higher perception threshold than the C/C genotype for 6-n-propyl-2-thiouracil (PROP) (p < .05), and caffeine (p < .05). The G-alleles had higher preferences for beer (OR: 6.25; p < .05), but lower for butter (OR: 0.64; p < .05) and cured meat (OR: 0.55; p < .05). Biochemical parameters and body composition markers did not differ between genotypes. Subjects with RS713598 polymorphism had a higher bitter taste perception threshold and higher or lower preferences for selected nutrient/energy dense foods, such as beer, butter and cured meat.


Timed release analgesics: Part 2. Nonopioids.
The problem of relieving patients' pain is a major challenge for health care professionals. New formulations of time-released pain medications offer exciting possibilities. However, there are cautions and patient education factors that are important when considering any timed-release product. This article looks at selected timed-release forms of nonopioids used for pain relief. Knowledge of the differences and similarities, benefits and cautions, and patient education factors will help the reader make safer and more informed decisions about prescribing and administering timed-release nonopioids. The first article in this two-part series (January/February 2001) examined narcotics.


Effects of mangrove removal on benthic organisms in the Siangshan Wetland in Hsinchu, Taiwan.
Mangrove degradation is a well-documented trend, but the spread of mangroves within the Siangshan Wetland in Hsinchu, Taiwan, runs contrary to that trend. The spread of mangroves changes the structure and functions of habitats for benthic organisms, causes infilling of estuaries and flooding and creates breeding grounds for small black mosquitoes. A large-scale mangrove-removal project was undertaken by the Hsinchu City Government from October 2015 to March 2016. They also investigated the consequences of mangrove removal on benthic organisms and adjacent habitats from October 2015 to September 2016, and the density, species count, Shannon-Wiener index (H'), and Pielou's evenness index (J') of the mangrove and non-mangrove regions were compared. In this study, we used satellite telemetry images to monitor fluctuations in mangrove density from 2006 to 2016. The non-mangrove region exhibited more variations than the mangrove region. After mangrove removal, species returned to their original habitats and noteworthy biological values significantly increased in the mangrove regions. This study presents evidence to argue that mangrove removal benefits benthic organisms. The results indicate that mangrove removal can be an appropriate habitat rehabilitation strategy for benthic organisms. The ecological findings of this study can inform coastal managers or other officials who seek to steward mangrove biomass.


Phosphoinositide control of membrane protein function: a frontier led by studies on ion channels.
Anionic phospholipids are critical constituents of the inner leaflet of the plasma membrane, ensuring appropriate membrane topology of transmembrane proteins. Additionally, in eukaryotes, the negatively charged phosphoinositides serve as key signals not only through their hydrolysis products but also through direct control of transmembrane protein function. Direct phosphoinositide control of the activity of ion channels and transporters has been the most convincing case of the critical importance of phospholipid-protein interactions in the functional control of membrane proteins. Furthermore, second messengers, such as [Ca(2+)]i, or posttranslational modifications, such as phosphorylation, can directly or allosterically fine-tune phospholipid-protein interactions and modulate activity. Recent advances in structure determination of membrane proteins have allowed investigators to obtain complexes of ion channels with phosphoinositides and to use computational and experimental approaches to probe the dynamic mechanisms by which lipid-protein interactions control active and inactive protein states.


Stable high-level expression of factor VIII in Chinese hamster ovary cells in improved elongation factor-1 alpha-based system.
Recombinant factor VIII (FVIII), used for haemophilia A therapy, is one of the most challenging among the therapeutic proteins produced in heterologous expression systems. Deletion variant of FVIII, in which the entire domain B is replaced by a short linker peptide, was approved for medical use. Efficacy and safety of this FVIII deletion variant are similar to full-length FVIII preparations while the level of production in CHO cells is substantially higher. Typical levels of productivity for CHO cell lines producing deletion variant FVIII-BDD SQ, described elsewhere, are 0.5-2 IU/ml, corresponding to the concentration of FVIII of about 0.2 μg/ml. Using standard vectors based on the cytomegalovirus promoter (CMV) and the dihydrofolate reductase cDNA we have previously obtained the cell line secreting 0.5 IU/ml of FVIII-BDD, which roughly corresponds to the previously published data. An expression system based on CHO genomic sequences including CHO-EEF1A promoter and Epstein-Barr virus terminal repeat fragment allowed us to achieve 80-fold increase in the production level as compared with the conventional expression system based on the CMV promoter. Immediately after the primary selection FVIII -producing cells secreted 5-10 IU/ml of FVIII-BDD, and after multi-stage methotrexate-driven amplification a stable clonal line 11A4H was selected, secreting 39 IU/ml of FVIII-BDD in the simple batch culturing conditions, which considerably exceeds known indicators for industrial producers of this protein. In contrast to other FVIII-BDD producing lines 11A4H accumulates low proportion of the secreted FVIII on the membrane. Its productivity may be further increased approximately two-fold by adding sodium butyrate and butylated hydroxyanisol to the culture medium. A five-stage purification process for the factor VIII was employed. It allowed isolation of the intact FVIII-BDD as was confirmed by mass spectrometry. Purified FVIII-BDD has a specific activity of 11,000 IU/mg, similar to known recombinant FVIII drugs. The recombinant FVIII-BDD was produced in CHO cells without addition of any animal-derived materials, purified and characterized. Novel genetic constructions for the expression of heterologous proteins combined with optimized cultivation method allowed to obtain the secretion level of biologically active recombinant FVIII increased by almost ten times as compared with the previously published analogues.


Outcome of segmental prosthesis reconstruction for diaphyseal bone tumors: a multi-center retrospective study.
The optimal reconstructive method after diaphyseal malignant bone tumor resection remains controversial. This multicenter clinical study was designed to investigate the clinical value and complications of segmental prosthesis in the repair of diaphyseal defects. We present 49 patients from three clinical centers treated with wide resection for primary or metastatic bone tumors involving the diaphysis of the femur, tibia, humerus, or ulna, followed by reconstruction using a modular intramedullary segmental prosthesis. Enrolled patients included 23 men and 26 women with a mean age of 63.3 years. Of these, seven patients had primary bone tumors and 42 patients had metastatic lesions. At the mean follow-up of 13.7 months, 17 patients were alive, 31 patients were deceased due to tumor progression, and one patient was dead of another reason. There were eight nononcologic complications (two with radial nerve injury, three with delayed incision healing, two with aseptic loosening in the proximal humerus prosthetic stem and one with structural failure) and three oncologic complications (three with primary tumor recurrence) among all patients. The incidence of complications in primary tumor patients (4/7, 57.1%) was higher than that in patients with metastatic tumors (7/42, 16.7%) (p = 0.036). Aseptic loosening and mechanical complications were not common for patients with primary tumors, although the reconstruction length difference was statistically significant (p = 0.023). No statistically significant differences were observed in limb function, while the mean musculoskeletal tumor society score was 21.2 in femora, 19.6 in humeri, and 17.8 in tibiae (p = 0.134). Segmental prostheses represent an optional method for the reconstruction of diaphyseal defects in patients with limited life expectancy. Segmental prostheses in the humerus experienced more complications than those used to treat lesions in the femur.


Phytochemicals Showing Antiangiogenic Effect in Pre-clinical Models and their Potential as an Alternative to Existing Therapeutics.
Angiogenesis, the formation of new blood vessels from a pre-existing vascular network, is an important hallmark of several pathological conditions, such as tumor growth and metastasis, proliferative retinopathies, including proliferative diabetic retinopathy and retinopathy of prematurity, age-related macular degeneration, rheumatoid arthritis, psoriasis, and endometriosis. Putting a halt to pathology-driven angiogenesis is considered an important therapeutic strategy to slow down or reduce the severity of pathological disorders. Considering the attrition rate of synthetic antiangiogenic compounds from the lab to reaching the market due to severe side effects, several compounds of natural origin are being explored for their antiangiogenic properties. Employing pre-clinical models for the evaluation of novel antiangiogenic compounds is a promising strategy for rapid screening of antiangiogenic compounds. These studies use a spectrum of angiogenic model systems that include HUVEC two-dimensional culture, nude mice, chick chorioallantoic membrane, transgenic zebrafish, and dorsal aorta from rats and chicks, depending upon available resources. The present article emphasizes the antiangiogenic activity of the phytochemicals shown to exhibit antiangiogenic behavior in these well-defined existing angiogenic models and highlights key molecular targets. Different models help to get a quick understanding of the efficacy and therapeutics mechanism of emerging lead molecules. The inherent variability in assays and corresponding different phytochemicals tested in each study prevent their immediate utilization in clinical studies. This review will discuss phytochemicals discovered using suitable preclinical antiangiogenic models, along with a special mention of leads that have entered clinical evaluation.


Microstructure and Quantitative Micromechanical Analysis of Wood Cell-Emulsion Polymer Isocyanate and Urea-Formaldehyde Interphases.
Emulsion polymer isocyanate (EPI) and urea-formaldehyde (UF) were selected as typical resin systems to investigate the microstructure of wood-adhesive interphases by fluorescence microscopy (FM) and confocal laser scanning microscopy (CLSM). Further, a quantitative micromechanical analysis of the interphases was conducted using nanoindentation. The FM results showed that the UF resin could penetrate the wood to a greater extent than the EPI resin, and that the average penetration depth for these two resin systems was higher in the case of latewood. CLSM allowed visualization of the resin distribution with contrasting colors, showing that the EPI resin could not penetrate the cell wall, whereas UF resin could enter the cell walls. The micromechanical properties of the cell walls were almost unaffected by EPI penetration but were significantly affected by UF penetration, especially in the first cell wall from the glueline. This further confirmed that only cell walls with resin penetration can improve the mechanical properties of the interphase regions.


Putting phylogeny into the analysis of biological traits: a methodological approach.
Phylogenetic comparative methods have long considered phylogenetic signal as a source of statistical bias in the correlative analysis of biological traits. However, the main life-history strategies existing in a set of taxa are often combinations of life history traits that are inherently phylogenetically structured. In this paper, we present a method for identifying evolutionary strategies from large sets of biological traits, using phylogeny as a source of meaningful historical and ecological information. Our methodology extends a multivariate method developed for the analysis of spatial patterns, and relies on finding combinations of traits that are phylogenetically autocorrelated. Using extensive simulations, we show that our method efficiently uncovers phylogenetic structures with respect to various tree topologies, and remains powerful in cases where a large majority of traits are not phylogenetically structured. Our methodology is illustrated using empirical data, and implemented in the adephylo package for the free software R.


Quantification of Pharmaceutical Compounds Based on Powder X-Ray Diffraction with Chemometrics.
We propose an approach for the simultaneous determination of multiple components in pharmaceutical mixed powder based on powder X-ray diffraction (PXRD) method coupled with chemometrics. Caffeine anhydrate, acetaminophen and lactose monohydrate were mixed at various ratios. The samples were analyzed by PXRD method in the ranges of 2θ=5.00-30.0 and 35.0-45.0 degrees. Obtained diffractograms were analyzed by conventional peak intensity method, multi curve resolution (MCR)-alternating least squares (ALS) method and partial least squares (PLS) method. Constructed PLS models can most accurately predict the concentrations among different methods used. Each regression vector of PLS correlates not only to the compound of interest but also to the coexisting compounds. The combination of PXRD and PLS methods is concluded to be powerful approach for analyzing multi components in pharmaceutical formulations.


Overcoming Challenges: Extending Cycle Life of Aqueous Zinc-Ion Batteries at High Zinc Utilization through a Synergistic Strategy.
Aqueous zinc-ion batteries (AZIBs) face challenges in achieving high energy density compared to conventional lithium-ion batteries (LIBs). The lower operating voltage and excessive Zn metal as anode pose constraints on the overall energy storage capacity of these batteries. An effective approach is to reduce the thickness of the Zn metal anode and control its mass appropriately. However, under the condition of using a thin Zn anode, the performance of AZIBs is often unsatisfactory. Through experiments and computational simulations, the electrode structural change and the formation of dead Zn as the primary reasons for the failure of batteries under a high Zn utilization rate are identified. Based on this understanding, a universal synergistic strategy that combines Cu foil current collectors and electrolyte additives to maintain the structural and thermodynamic stability of the Zn anode under a high Zn utilization rate (ZUR) is proposed. Specifically, the Cu current collectors can ensure that the Zn anode structure remains intact based on the spontaneous filling effect, while the additives can suppress parasitic side reactions at the interface. Ultimately, the symmetric cell demonstrates a cycling duration of 900 h at a 70% ZU, confirming the effectiveness of this strategy.


THE EFFECTS OF GENE FLOW ON REINFORCEMENT.
We explore the possibility that differences in the pattern of gene flow between populations may affect the evolution of reinforcement by comparing pairs of populations undergoing one-way migration versus symmetric migration. The case of symmetric migration is modeled by a two-island model, where the two populations exchange equal proportions of migrants each generation. One-way migration is modeled by a continent-island model, where migration is in one direction from a large continental population with a fixed genotype to an island population whose genotype frequencies can vary. Hybrid inviability is assumed to be caused by epistatic interactions between background loci. We examine the spread of an introduced preference allele for a previously unpreferred male trait that characterizes one of the populations. Computer simulations indicate that with a weak introduced preference, reinforcement is possible under a wide range of parameter values in a symmetric migration model but cannot occur in a one-way migration model. Reinforcement with one-way migration can occur only with a very strong introduced preference and very strong selection against hybrids. Our results suggest that the speciation of a peripheral isolate, which undergoes essentially one-way migration, may be difficult to complete if secondary contact occurs before reproductive isolation is fully developed.


Utility of exhaled nitric oxide as a noninvasive biomarker of lung inflammation in a disease model.
There is a great deal of interest in developing less invasive markers for monitoring airway inflammation and the effect of possible novel anti-inflammatory therapies that may take time to impact on disease pathology. Exhaled nitric oxide (eNO) has been shown to be a reproducible, noninvasive indicator of the inflammatory status of the airway in the clinic. The aim of the present study was to determine the usefulness of measuring eNO as a marker of the anti-inflammatory impact of glucocorticoid and an inhibitor of kappaB kinase-2 (IKK-2) inhibitor 2-[(aminocarbonyl)amino]-5-(4-fluorophenyl)-3-thiophenecarboxamide (TPCA-1), in a pre-clinical model of airway inflammation. Rats were given vehicle, budesonide or TPCA-1 prior to exposure to lipopolysaccharide, previously shown to induce an increase in eNO and airway neutrophilia/eosinophilia. Comparison of the effect of the two compounds on inflammatory components demonstrated a significant correlation between the impact on eNO and inflammatory cell burden in the airway. The current study demonstrates the usefulness of profiling potential disease-modifying therapies on exhaled nitric oxide levels and the way in which an effect on this noninvasive biomarker relates to effects on pathological parameters such as lung cellularity. Information from studies such as the current one would suggest that the measurement of exhaled nitric oxide has potential for monitoring inflammatory status in lung tissue.


Effect of liver pâté enrichment with flaxseed oil and flaxseed extract on lipid composition and stability.
There is an increase interest from food technologists in the improvement of the nutritional value of meat products, especially their lipid profile and shelf life. The aim of this study was to determine the effect of animal fat replacement with flaxseed oil (20%) and flaxseed extract addition (0.05%-0.25%) on changes in composition and stability of liver pâté lipid fractions. Replacement of animal fat with flaxseed oil was observed to lower the saturated and monoenoic fatty acid in the product's fatty acid profile by around 12% and to raise the polyene fatty acid contribution by more than 70% when compared to the control. The replacement also resulted in enriching the pâté with phytosterols (β-sitosterol, campesterol, and cycloartenol). The addition of 0.05% and 0.10% ethanol flaxseed extract significantly slowed down lipid oxidation changes during storage of the pâté. The study showed that a storage-stable meat product - liver pâté - characterized by a high nutritional value can be designed through the addition of flax oil rich in unsaturated fatty acids and flaxseed extract. The addition of 0.05% of the extract was found to be sufficient to improve the liver pâté's oxidative stability; it can thus be recommended for this type of product. © 2018 Society of Chemical Industry.


Do people serve as cues to smoke?
Recent research has identified that the environments in which smoking has previously occurred can alone, in the absence of any explicit smoking stimuli (e.g., cigarettes, lighters), serve as cues that induce robust craving to smoke. The goal of the present study was to determine if people can similarly function as smoking and nonsmoking cues capable of directly affecting smokers' cue-induced craving. Smokers (N = 72) borrowed cameras to take photos of the people in their lives around whom they do and do not smoke ("personal" smoking and nonsmoking people, PS and PN, respectively). Self-report and physiological cue reactivity to those photos were compared with smokers' reactivity to photos of people unknown to them ("standard" smoking and nonsmoking people, SS and SN, respectively). Results suggest that the people around whom smokers regularly smoke (PS) can alone function as cues capable of eliciting patterns of reactivity similar to that evoked by proximal and environment smoking cues, namely, increased craving to smoke, negative affect, and excitement. In contrast, the people around whom smokers do not smoke become associated with not smoking (PN) and serve a potential protective function by reducing craving and increasing calm. This novel investigation and its results have implications for promoting smoking cessation by developing strategies to manage a smoker's social environment.


Selective neck dissection following adjuvant therapy for advanced head and neck cancer.
In the past, surgeons believed that in order to eradicate regional disease, a radical or modified radical neck dissection was necessary. An evolution in surgical principles and the popularization of primary chemoradiation has raised the questions regarding the role of neck dissection and the extent of neck dissection following therapy. The aim of this study was to determine the efficacy of selective neck dissection (SND) for patients with N2 or N3 disease following treatment with primary radiation therapy or chemoradiation. A retrospective review of 58 patients with stage III or IV head and neck squamous cell carcinoma was conducted. The primary sites included base of tongue (n = 15), hypopharynx (n = 12), tonsil (n = 16), larynx (n = 11), and unknown primary (n = 4). Definitive treatment consisted of either concomitant chemoradiation (67.2%) or external beam radiation therapy (32.8%). In the monotherapy group, all patients received a total curative dose of 66 to 72 Gy in once-daily fractions of 180 to 200 cGy. The combined chemoradiation group received a similar radiation schedule and a 4-day continuous infusion of cisplatin (20 mg/m(2)/day) and 5-fluorouracil (1000 mg/m(2)/day). A planned SND of levels II to IV was performed on all the patients 3 to 6 weeks after completion of definitive medical therapy. Seventy neck dissections were performed on 58 patients with advanced neck disease following radiation or chemoradiation. The median time of follow-up was 34 months (range, 9-71 months) following the neck dissection. Pathologically, 22.4% (13/58) of the patients had viable tumor cells identified in the neck dissection specimen. Seventy-two percent of the patients are currently alive, and 28% died as a result of distant disease, local or regional recurrence, or other causes. Of patients who died from distant disease, 11% had pathological evidence of residual tumor cells identified in the neck dissection specimen. Of patients who died from local or regional disease, 50% had pathological evidence of residual tumor cells identified in the neck dissection specimen. The rate of regional recurrence following SND is similar to reported rates following modified/radical neck dissection. This suggests that SND provides an appropriate surgical option for advanced neck disease in select patients following adjuvant therapy.


BRAF inhibitor vemurafenib improves the antitumor activity of adoptive cell immunotherapy.
Combining immunotherapy with targeted therapy blocking oncogenic BRAFV600 may result in improved treatments for advanced melanoma. In this study, we developed a BRAFV600E-driven murine model of melanoma, SM1, which is syngeneic to fully immunocompetent mice. SM1 cells exposed to the BRAF inhibitor vemurafenib (PLX4032) showed partial in vitro and in vivo sensitivity resulting from the inhibition of MAPK pathway signaling. Combined treatment of vemurafenib plus adoptive cell transfer therapy with lymphocytes genetically modified with a T-cell receptor (TCR) recognizing chicken ovalbumin (OVA) expressed by SM1-OVA tumors or pmel-1 TCR transgenic lymphocytes recognizing gp100 endogenously expressed by SM1 resulted in superior antitumor responses compared with either therapy alone. T-cell analysis showed that vemurafenib did not significantly alter the expansion, distribution, or tumor accumulation of the adoptively transferred cells. However, vemurafenib paradoxically increased mitogen-activated protein kinase (MAPK) signaling, in vivo cytotoxic activity, and intratumoral cytokine secretion by adoptively transferred cells. Taken together, our findings, derived from 2 independent models combining BRAF-targeted therapy with immunotherapy, support the testing of this therapeutic combination in patients with BRAFV600 mutant metastatic melanoma.


Comparative analysis of protein unfoldedness in human housekeeping and non-housekeeping proteins.
Absence of any regular structure is increasingly being observed in structural studies of proteins. These disordered regions or random coils, which have been observed under physiological conditions, are indicators of protein plasticity. The wide variety of interactions possible due to the flexibility of these 'natively disordered' regions confers functional advantage to the protein and the organism in general. This concept is underscored by the increasing proportion of intrinsically unstructured proteins seen with the ascension in the complexity of the organisms. The 'natively unfolded/disordered' state of the protein can be predicted utilizing Uversky's or Dunker's algorithm. We utilized Uversky's prediction scheme and based on the unique position of a protein in the charge-hydrophobicity plot, a derived net score was used to predict the overall disorder of the human housekeeping and non-housekeeping proteins. Substantial numbers of proteins in both the classes were predicted to be unfolded. However, comparative genomic analysis of predicted unfolded Homo sapiens proteins with homologues in Caenorhabditis elegans, Drosophila melanogaster and Mus musculus revealed significant increase in unfoldedness in non-housekeeping proteins in comparison with housekeeping proteins. Our analysis in the evolutionary context suggests addition or substitution of amino acid residues which favour unfoldedness in non-housekeeping proteins compared to housekeeping proteins.


Amino acid-induced responses in CNS neurons of normal mouse and Wriggle Mouse Sagami.
Amino acid-induced responses and their regional heterogeneities in the motor cortical and hippocampal (CA1) neurons of normal mouse and Wriggle Mouse Sagami (WMS) were studied by conventional patch-clamp techniques. There were no differences in the current densities and EC50s of glutamate-, gamma-amino butyric acid (GABA)-, and glycine-induced responses between the two groups of mice. In both types of mice, the GABA-induced current density of cortical neurones was higher and EC50 was lower than in hippocampal neurones, whereas the responses induced by glycine and glutamate did not show any differences.


Humidity-Responsive Photonic Crystals with pH and SO2 Gas Detection Ability Based on Cholesteric Liquid Crystalline Networks.
Dynamic photonic crystals with tunable structural colors have been a hot topic in the research of anticounterfeiting devices, decoration, and detection. In this work, we prepared cholesteric liquid crystalline network (CLCN)-based photonic crystals that present humidity- and SO2 gas-responsive behaviors. The covalently cross-linked CLCN film presents humidity-responsive color changes due to the swelling/deswelling of the matrix under different humidity conditions. When treating the CLCN film with SO2 gas, the carboxylic salt converted to the acid and the film was not able to respond to the humidity change anymore. The mechanism of the SO2 gas-gated humidity responsiveness of the CLCN film was characterized. It was found that the acidic gas caused changes of pH, resulting in the conversion of the salt to acid and alteration of the surface property. The influence of concentration of SO2 gas and pH on humidity responsiveness of the CLCN film was investigated. We hope that this method provides inspirations for the design and fabrication of visualized pH and acidic gas detectors.


Differential expression of Notch1 intracellular domain and p21 proteins, and their clinical significance in gastric cancer.
Changes in the expression of the Notch1 intracellular domain (NICD) and p21 proteins have been shown to be closely associated with the development and progression of a number of cancers. The present study aimed to investigate the expression levels of the two proteins in gastric carcinoma and precancerous lesions, and to determine the clinical significance of this. A total of 109 gastric cancer, 57 precancerous gastric lesion, 50 chronic superficial gastritis and 17 normal gastric mucosa patients were recruited for immunohistochemical staining of NICD and p21 protein expression. The protein expression levels in the gastric cancer patient samples were associated with the clinicopathological and survival data. NICD protein levels were upregulated gradually from normal gastric mucosae through chronic superficial gastritis and precancerous gastric lesions to gastric cancer tissues (P<0.01), whereas p21 protein levels were downregulated accordingly (P<0.01). Increased NICD and a loss of p21 expression were closely associated with tumor dedifferentiation, depth of tumor invasion, lymph node metastasis, surface morphology and Lauren classification in gastric cancer. Thus, NICD expression was inversely associated with p21 expression. In addition, the overall survival rate was greater in NICD- and P21+ patients than in NICD+ and P21- patients, respectively (P<0.05). The COX regression multivariate analysis revealed that NICD+, p21-, depth of tumor invasion and lymph node metastasis were all independent prognostic factors for patients with gastric cancer. NICD and p21 proteins are differentially expressed in gastric cancer and the aberrant expression of these proteins is associated with an advanced tumor stage, tumor metastasis and overall patient survival. Future studies are required to further evaluate the two proteins as novel prognostic markers for patients with gastric cancer.


Heart transplantation for end-stage heart failure caused by iron overload.
Few reports exist concerning heart transplantation in recipients with end-stage myocardiopathy-associated heart failure caused by iron overload occurring with beta-thalassaemia, Diamond-Blackfan syndrome or haemochromatosis. Seven potential transplant candidates (six male, one female, mean age 26 years) with such heart failure, following desferrioxamine application subcutaneously over a number of years, and intravenously during their hospitalization before transplantation, were retrospectively analysed. Five were New York Heart Association (NYHA) class IV, three experienced one or more resuscitations immediately before transplantation could be performed. Continuous, high-volume, veno-venous haemofiltration was necessary in two patients. One of these two candidates additionally had to be bridged, first with a right ventricular, then with a biventricular assist device. Five of the seven patients survived, two with haemochromatosis, one with beta-thalassaemia major and one with Diamond-Blackfan syndrome following transplantation. One non-transplanted candidate with beta-thalassaemia major has been recompensated for 5 years. Survival was 14-74 months. Our results demonstrate the feasibility and indication of transplantation in patients with such heart failure and the satisfying outcome of immunosuppression is described.


Effect of Different Cooking Methods on the Bioactive Components, Color, Texture, Microstructure, and Volatiles of Shiitake Mushrooms.
The effects of different cooking methods (steaming, boiling, air frying, and oven baking) and cooking times (0, 5, 10, 15, and 20 min) on the bioactive components (total phenol, total flavonoid, crude polysaccharides, and eritadenine), color, texture, microstructure, and volatiles in shiitake mushrooms were investigated in this study. Steaming, boiling, and air frying for 5-20 min could decrease the contents of all the four bioactive components in the shiitake mushroom. However, oven baking for 5 min and 10 min showed the highest contents of total phenolics and total flavonoids, respectively. Moreover, the lowest losses of crude polysaccharides and eritadenine were observed for oven baking for 5 min and 15 min, respectively. The lightness of shiitake mushrooms was decreased by all treatments; however, steaming could keep a higher brightness compared with other methods. The microstructure was damaged by all cooking methods, especially air frying for 20 min. Meanwhile, steaming for 20 min decreased the hardness mostly, and there was no significant difference with air frying for 20 min. All cooking treatments decreased the complexity of the flavors and the relative contents of volatile compounds; the lowest contents were found when boiling for 5 min. From these results it can be seen that the physical, histological, and chemical features in shiitake mushroom were influenced by cooking methods and times. In addition, our results provide valuable information for the cooking and processing of shiitake mushrooms and other fungi.


[Implementation of a telediagnostic system for tuberculosis and determination of multi-drug resistance based in the MODS method in Trujillo, Peru].
To implement a system for remote diagnosis of tuberculosis and multidrug resistance (MDR) using the Microscopic-Observation Drug Susceptibility Assay (MODS) method in the Mycobacteria Laboratory, Trujillo Center of Excellence in Tuberculosis (CENEX-Trujillo). The system included a variant of an algorithm for recognition of Mycobacterium tuberculosis recently reported from digital images of MODS cultures of sputum samples. The recognition algorithm was optimized using a retraining statistical model based on digital images of MODS cultures from CENEX-Trujillo. Images of 50 positive MODS cultures of patients with suspected multidrug-resistant tuberculosis were obtained between January and October 2012 in the CENEX-Trujillo. The sensitivity and specificity to recognize strings of tuberculosis were 92.04% and 94.93% respectively using objects. The sensitivity and specificity to determine a positive tuberculosis field were 95.4% and 98.07% respectively using pictures. The results demonstrated the feasibility of the implementation of telediagnostics in remote locations, which may contribute to the early detection of multidrug-resistant tuberculosis by MODS method.


Graftless sinus augmentation with simultaneous dental implant placement: clinical results and biological perspectives.
After a sinus lifting procedure, the compartment around the implants under the sinus mucosal lining in the sinus floor is filled with a blood clot from surrounding bleeding. The aim of this study was to evaluate the feasibility of bone formation following graftless sinus lifting with the simultaneous placement of dental implants. Thirty graftless sinus lifting procedures were performed and 72 dental implants placed in 18 consecutive patients, using the lateral window approach. Clinical and radiological follow-up was conducted throughout the 6-month healing period. Biopsies of 30 cases were collected at 6 months post-treatment: 15 biopsies were taken from the newly formed bone near the basal floor and 15 from the newly formed bone near the elevated membrane. New bone consolidation in the maxillary sinus was apparent radiologically and histologically at 6 months after sinus augmentation, providing an average 6.14±1.34mm of bone-gain. Based on histological analysis and histomorphometric data, the consolidated bone in the augmented sinus comprised 56.7±11.9% to 59.9±13.4% vital bone tissue. Out of the 72 implants placed, only four failed, indicating a 94% overall implant survival rate. Based on this case series, blood clot can be considered autologous osteogenic graft material, to which osteoprogenitors can migrate, differentiate, and regenerate bone.


The nucleotide sequence of the genes, fanE and fanF of Escherichia coli K99 fimbriae.
K99 fimbriae of enterotoxigenic Escherichia coli consist of eight different subunits. A major subunit called fimbrillin forms fimbrial structure and a minor subunit called adhesin localizes at the tip of fimbriae and recognizes host receptor ganglioside. Within this eight gene cluster, fanE and fanF have not yet been sequenced. In this study, fanE and fanF genes were sequenced by analyzing several DNA fragments produced by endonuclease or exonuclease digestion. The fanE gene encoded 227 amino acids containing 20 amino acids of signal peptide starting from GTG (valine) and showed a homology to fanA-fanB. The fanF gene encoded 271 amino acids containing 20 amino acids of signal peptide starting from ATG (methionine) and showed homologies to the fanD gene, fimbrillin gene of F41, adhesin gene of P fimbriae (papG) and adhesin gene of Type 1 fimbriae (fimH). E and F subunits had fifteen and fourteen hydrophobic domains, respectively, which periodically appeared possibly forming a hydrophobic region.


Isolation of an Adoxophyes orana granulovirus (AdorGV) occlusion body morphology mutant: biological activity, genome sequence and relationship to other isolates of AdorGV.
A granulovirus (GV) producing occlusion bodies (OBs) with an unusual appearance was isolated from Adoxophyes spp. larvae in the field. Ultrastructural observations revealed that its OBs were significantly larger and cuboidal in shape, rather than the standard ovo-cylindrical shape typical of GVs. N-terminal amino acid sequence analysis of the OB matrix protein from this virus suggested that this new isolate was a variant of Adoxophyes orana granulovirus (AdorGV). Bioassays of this GV (termed AdorGV-M) and an English isolate of AdorGV (termed AdorGV-E) indicated that the two isolates were equally pathogenic against larvae of Adoxophyes honmai. However, AdorGV-M retained more infectivity towards larvae after irradiation with UV light than did AdorGV-E. Sequencing and analysis of the AdorGV-M genome revealed little sequence divergence between this isolate and AdorGV-E. Comparison of selected genes among the two AdorGV isolates and other Japanese AdorGV isolates revealed differences that may account for the unusual OB morphology of AdorGV-M.


Rapid quantification of adult and developing mouse spatial vision using a virtual optomotor system.
To develop a simple, rapid method of quantifying the spatial vision of mice. A rotating cylinder covered with a vertical sine wave grating was calculated and drawn in virtual three-dimensional (3-D) space on four computer monitors facing to form a square. C57BL/6 mice standing unrestrained on a platform in the center of the square tracked the grating with reflexive head and neck movements. The spatial frequency of the grating was clamped at the viewing position by repeatedly recentering the cylinder on the head. Acuity was quantified by increasing the spatial frequency of the grating until an optomotor response could not be elicited. Contrast sensitivity was measured at spatial frequencies between 0.03 and 0.35 cyc/deg. Grating acuity was measurable on the day of eye opening (postnatal day [P]15: mean acuity, 0.031 cyc/deg) and reached a maximum (approximately 0.4 cyc/deg) by P24. A peak in the contrast sensitivity function emerged on P16 (4.7, or 21% contrast at 0.064 cyc/deg). The peak remained at 0.064 cyc/deg and climbed to a maximum sensitivity of 24.5, or 4% contrast, by P29. Acuity was obtained in each mouse in <10 minutes, and a detailed contrast sensitivity curve was generated in approximately 30 minutes. The virtual optomotor system provides a simple and precise method for rapidly quantifying mouse vision. Behavioral measures of vision in mice are essential for interpreting the results of experiments designed to reveal the cellular and molecular mechanisms of vision and visual development and for evaluating potential treatments for visual diseases.


Cost-effectiveness of screening for primary aldosteronism in hypertensive patients in Australia: a Markov modelling analysis.
Primary aldosteronism affects 3-14% of hypertensive patients in the primary care setting and up to 30% in the hypertensive referral units. Although primary aldosteronism screening is recommended in patients with treatment-resistant hypertension, diagnosis at an earlier stage of disease may prevent end-organ damage and optimize patient outcomes. A Markov model was used to estimate the cost-effectiveness of screening for primary aldosteronism in treatment and disease (cardiovascular disease and stroke) naive hypertensive patients. Within the model, a 40-year-old patient with hypertension went through either the screened or the unscreened arm of the model. They were followed until age 80 or death. In the screening arm, the patient underwent standard diagnostic testing for primary aldosteronism if the screening test, aldosterone-to-renin ratio, was elevated above 70 pmol/l : mU/l. Diagnostic accuracies, transition probabilities and costs were derived from published literature and expert advice. The main outcome of interest was the incremental cost effectiveness ratio (ICER). Screening hypertensive patients for primary aldosteronism compared with not screening attained an ICER of AU$35 950.44 per quality-adjusted life year (QALY) gained. The results were robust to different sensitivity analyses. Probabilistic sensitivity analysis demonstrated that in 73% of the cases, it was cost-effective to screen at the commonly adopted willingness-to-pay (WTP) threshold of AU$50 000. The results from this study demonstrated that screening all hypertensive patients for primary aldosteronism from age 40 is cost-effective. The findings argue in favour of screening for primary aldosteronism before the development of severe hypertension in the Australian healthcare setting.


[Application and analysis of ethnic medicines in "Three Minorities" in Inner Mongolia].
This paper investigates and counts the ethnic medicines of the Ewenki,Daur and Oroqen ethnic groups,which are known as the " Three Minorities" in Inner Mongolia. Through the methods of literature collection,interview investigation,and resource investigation,different ethnic medicines were collected on the main diseases,drug varieties,drug-injection sites,and drug administration methods. Through data statistics and SPSS analysis,the similarities and individual differences between the three ethnic groups were clarified. The results indicated the predicament of the current national medicine,which is helpful for the protection and inheritance of ethnic medicine.


Glucocorticosteroids inhibit leukotriene production.
The mode of action of corticosteroids, important drugs in the treatment of inflammatory disease, is not yet fully understood. Corticosteroids are known to inhibit phospholipase A2 in unprimed eosinophils and basophils, preventing leukotriene synthesis, but their effect on cells that are already primed is unknown. As inflammatory cells from atopic subjects are often primed in vivo, we studied the effects of two potent corticosteroids on basophil sulfidoleukotriene production in peripheral blood mixed leukocytes (PBML) from in-season and out-of-season atopic individuals. Cells were incubated for 24 hours with mometasone furoate or beclomethasone dipropionate, primed with IL-3, stimulated with calcium ionophore, buffer, allergen or anti-IgE, and leukotriene production was quantified. Peripheral blood mononuclear leukocytes from five of ten donors (in season) produced elevated sulfidoleukotrienes without a stimulus; cells from seven donors responded to anti-IgE by increased sulfidoleukotrienes. Neither steroid consistently affected sulfidoleukotriene production in anti-IgE-stimulated cells which were releasing sulfidoleukotrienes in the absence of a stimulant. In comparison, sulfidoleukotriene production was significantly reduced by 0.01 to 10 nM beclomethasone dipropionate or mometasone furoate when the cells were primed with IL-3 after exposure to the drug and stimulated with calcium ionophore or allergen, but no dose-relationship was apparent. Leukotriene production by PBML in response to anti-IgE was potently inhibited by all concentrations of mometasone furoate (0.01 nM to 1 microM) with an inhibitory concentration50 of less than 0.01 nM. Beclomethasone dipropionate inhibited sulfidoleukotriene production in this group (inhibitory concentration50 6 nM) in a dose-dependent manner. Sulfidoleukotriene production and, conceivably, priming may be more effectively inhibited by mometasone furoate than beclomethasone dipropionate.


Roles of adrenomedullin 2 in regulating the cardiovascular and sympathetic nervous systems in conscious rats.
Recently, a new member of the calcitonin gene-related peptide (CGRP) family, adrenomedullin 2 (AM2) or intermedin (IMD), was identified. AM2/IMD has been shown to have a vasodilator effect in mice and rats and an effect on urine formation in rats. In the present study, we investigated the effects of intravenously infused rat AM2 (rAM2) on blood pressure (BP), heart rate (HR), renal sympathetic nerve activity (RSNA), and renal blood flow (RBF) in conscious unrestrained rats relative to the effects of rat adrenomedullin (rAM) and proadrenomedullin NH2-terminal 20 peptide (rPAMP). Intravenous infusion of rAM2 (5 nmol/kg) significantly decreased BP and increased HR, RSNA, and RBF. These hypotensive and sympathoexcitatory effects diminished after 20 min, and HR returned to control levels 30 min after cessation of the infusion. In contrast, a significant increase in RBF was still evident 60 min after cessation of the peptide infusion. The duration of BP, HR, and RSNA responses was longer with rAM (5 nmol/kg) than with rAM2 infusion, whereas the increases in RBF induced by rAM2 and rAM were similar in their amplitude and duration. Infusion of rPAMP (200 nmol/kg) increased HR and RSNA but had no effect on RBF. Baroreceptor denervation suppressed, but did not diminish, the increases in HR and RSNA to rAM2. These findings indicate that the physiological roles of rAM2 and rAM are similar and that rAM2 also has a long-lasting vasodilator action on the renal vascular bed.


A recombinant human interferon-alpha B/D hybrid with a broad host-range.
A recombinant interferon (IFN) hybrid has been found to have a broad host-range of activity in an antiviral assay (plaque reduction of vesicular stomatitis virus) and also high efficacy as an antiviral agent in at least 12 different animal cell species. The IFN hybrid consists of amino acids 1 to 60 from HuIFN-alpha B and amino acids 61 to 166 from HuIFN-alpha D. The profile of cross-species activity of the IFN-alpha B/D hybrid has been compared with that of HuIFN-alpha F, and of the parents HuIFN-alpha B and -alpha D. When both IFN-alpha B and -alpha D were active in a cell species, the hybrid IFN had comparable or better activity than the more active parental IFN. The hybrid shared a broad cross-species activity with IFN-alpha D. However, the IFN-alpha B/D hybrid was 10-fold more active on human cells, 30-fold more active on rabbit cells, and 50-fold more active on mouse cells than IFN-alpha D.


mRNA deadenylation by PARN is essential for embryogenesis in higher plants.
Deadenylation of mRNA is often the first and rate-limiting step in mRNA decay. PARN, a poly(A)-specific 3' --> 5' ribonuclease which is conserved in many eukaryotes, has been proposed to be primarily responsible for such a reaction, yet the importance of the PARN function at the whole-organism level has not been demonstrated in any species. Here, we show that mRNA deadenylation by PARN is essential for viability in higher plants (Arabidopsis thaliana). Yet, this essential requirement for the PARN function is not universal across the phylogenetic spectrum, because PARN is dispensable in Fungi (Schizosaccharomyces pombe), and can be at least severely downregulated without any obvious consequences in Metazoa (Caenorhabditis elegans). Development of the Arabidopsis embryos lacking PARN (AtPARN), as well as of those expressing an enzymatically inactive protein, was markedly retarded, and ultimately culminated in an arrest at the bent-cotyledon stage. Importantly, only some, rather than all, embryo-specific transcripts were hyperadenylated in the mutant embryos, suggesting that preferential deadenylation of a specific select subset of mRNAs, rather than a general deadenylation of the whole mRNA population, by AtPARN is indispensable for embryogenesis in Arabidopsis. These findings indicate a unique, nonredundant role of AtPARN among the multiple plant deadenylases.


[Action of neurotensin on duodenal alkaline secretion in rats. Comparison with pancreatic and gastric secretion].
Neurotensin is released from endocrine N cells of the ileum after meals, and might take part in the regulation of bicarbonate secretion by the pancreas and duodenum. The aim of this study was to define the effect of neurotensin on duodenal bicarbonate secretion, in comparison with its effect on gastric and pancreatic secretions. Neurotensin produced a dose-related increase of duodenal bicarbonate secretion with an ED50 of 60 pmol/kg.h. The maximal effect (about 2 times the basal level) was observed with 600 pmol/kg.h. Equimolar doses (600 pmol/kg.h) of xenopsin, neuromedin N, neurotensin 8-13 produced the same effect as neurotensin 1-13. Neurotensin fragments 1-11 and 9-13 (600 pmol/kg.h) had no significant effect on duodenal bicarbonate secretion. Indomethacin, atropine, naloxone, or the CCK antagonist L364,718 had no effect on neurotensin-stimulated bicarbonate secretion. Hexamethonium and vagotomy reduced the neurotensin effect by about 50 percent (P less than 0.05). Neurotensin produced a dose-related increase of pancreatic bicarbonate secretion with an ED50 of 150 pmol/kg.h, a decrease of gastric acid secretion with an ED50 of 2,400 pmol/kg.h, and a decrease of gastric pepsin secretion with an ED50 of 2,760 pmol/kg.h. This study shows that neurotensin stimulates duodenal bicarbonate secretion in doses which may be physiological. This biological activity depends on the presence of the C-terminal 8-13 fragment. The mechanism is complex, and depends, for approximately half, on vagal fibers (sensitive or motor), nicotinic synapses, and a non cholinergic effector. The other half of the effect, still unexplained, could be a direct effect on mucosal cells. Pancreatic and duodenal bicarbonate secretions were more sensitive to neurotensin than gastric secretion (acid and pepsin).


Rapid growth of liver hemangioma following interferon treatment for hepatitis C in a young woman.
Liver hemangioma is the most common benign liver tumor. Giant hemangiomas can grow and give various clinical symptoms. We describe a case of rapid growth of liver hemangioma in a seventeen-year-old girl. She was treated with interferon alpha 2b for hepatitis C. About 12 months later she was admitted to the hospital with symptoms of large epigastric mass, anemia and thrombocytopenia. Ultrasound scan and computed tomography confirmed the clinical picture, suggesting giant liver hemangioma. During surgery large cavernous hemangioma deeply penetrating into normal liver parenchyma was found. In view of recent hepatitis C infection the resection procedure (left extended hemihepatectomy) was assessed as being dangerous for the patient's life. Ligation of the left branches of hepatic artery and portal vein reducing tumor blood supply was performed. The postoperative period was uneventful; the hemangioma in the left lobe had shrunk. Ultrasound and CT scans performed one and two years after surgery showed that the size of the hemangioma remained stable and the patient was asymptomatic. In this described case three potential factors stimulating the growth of hemangioma (puberty, hepatitis C infection and interferon treatment) are discussed. The potential role of vascular endothelial growth factor and fibroblast growth factor is raised.


Associations between Weight-Adjusted Waist Index and Abdominal Fat and Muscle Mass: Multi-Ethnic Study of Atherosclerosis.
The weight-adjusted waist index (WWI) reflected body compositional changes with aging. This study was to investigate the association of WWI with abdominal fat and muscle mass in a diverse race/ethnic population. Computed tomography (CT) data from 1,946 participants for abdominal fat and muscle areas from the Multi-Ethnic Study of Atherosclerosis (785 Whites, 252 Asians, 406 African American, and 503 Hispanics) were used. Among them, 595 participants underwent repeated CT. The WWI was calculated as waist circumference (cm) divided by the square root of body weight (kg). The associations of WWI with abdominal fat and muscle measures were examined, and longitudinal changes in abdominal composition measures were compared. In all race/ethnic groups, WWI was positively correlated with total abdominal fat area (TFA), subcutaneous fat area, and visceral fat area, but negatively correlated with total abdominal muscle area (TMA) and abdominal muscle radiodensity (P<0.001 for all). WWI showed a linear increase with aging regardless of race and there were no significant differences in the WWI distribution between Whites, Asians, and African Americans. In longitudinal analyses, over 38.6 months of follow-up, all abdominal fat measures increased but muscle measures decreased, along with increase in WWI. The more the WWI increased, the more the TFA increased and the more the TMA decreased. WWI showed positive associations with abdominal fat mass and negative associations with abdominal muscle mass, which likely reflects the abdominal compositional changes with aging in a multi-ethnic population.


Activation of Peroxisome Proliferator-Activated Receptor Gamma and Disruption of Progesterone Synthesis of 2-Ethylhexyl Diphenyl Phosphate in Human Placental Choriocarcinoma Cells: Comparison with Triphenyl Phosphate.
2-Ethylhexyl diphenyl phosphate (EHDPP), an organophosphate flame retardant (OPFR), is frequently detected in human blood. In this study, the sensitive dual-luciferase reporter gene assay and molecular docking were used to investigate the activation of EHDPP to human peroxisome proliferator-activated receptor gamma (PPARG). Results show that EHDPP exhibited stronger PPARG activation (EC20: 2.04 μM) than triphenyl phosphate (TPhP) (EC20: 2.78 μM). EHDPP upregulated the gene expression of 3β-hydroxysteroid dehydrogenase type 1 (3β-HSD1) in human placental choriocarcinoma cells in a dose-dependent manner, and the lowest observable effective concentration was 10 μM, lower than that of TPhP (20 μM). EHDPP significantly altered progesterone secretion at a lower concentration (10 μM) than that of TPhP (20 μM), and both EHDPP and TPhP significantly promoted human chorionic gonadotropin (hCG) production at 20 μM. Furthermore, inactivation of PPARG by either a pharmacological inhibitor (GW9662) or small interfering RNA (siRNA) abolished the change in progesterone secretion and gene expression in the cells exposed to EHDPP, suggesting that the PPARG signaling pathway plays a role in the upregulation of progesterone by the two OPFRs. This is the first report to show that OPFRs can alter the biosynthesis of progesterone in the placenta, which could affect female reproduction and fetal development.


Piperazine amide linker for cyclative cleavage from solid support: traceless synthesis of dihydroquinoxalin-2-ones.
A piperazine amide linker for cyclative cleavage from solid support and its use in the traceless solid-phase synthesis of dihydroquinoxalinones are described. Piperazine was attached to Wang resin via a carbamate linkage and acylated with Fmoc-amino acids. Following Fmoc group removal, resin-bound amines were reacted with 1-fluoro-2-nitrobenzenes. The nitro group of the resulting 2-nitroanilines was reduced, and acyclic precursors, in contrast to traditional ester-type linkage, remained attached to the resin. Target dihydroquinoxalinones were obtained either by acid- or microwave-mediated cyclative cleavage. The synthesis provided crude compounds of high purity and enabled the preparation of stable immobilized linear intermediates. The linker is suitable for combinatorial synthesis of compound libraries.


Japanese Patients' and Physicians' Preferences for Anticoagulant Use in Atrial Fibrillation: Results from a Discrete-choice Experiment.
Background: Anticoagulants are recommended for stroke prevention in patients with atrial fibrillation (AF), but are associated with an increased risk of bleeding; therefore, physicians face benefit-risk tradeoffs when prescribing anticoagulants to AF patients. Although the unmet medical need for safer anticoagulants has been well-documented, there is limited information about the importance that patients and physicians place on cardiovascular events. Objectives: The aim of this study was to quantify patients' and physicians' willingness to accept tradeoffs between the benefits and risks of anticoagulants in order to 1) document the potential differences between patients' and physicians' perceptions of benefits and risks and 2) support physicians' clinical decision making. Methods: Preferences from Japanese AF patients and board-eligible or board-certified physicians were elicited using a discrete-choice experiment. Random-parameters logit models were used to estimate importance weights for treatment-related changes in the annual risk of stroke, myocardial infarction, embolism, and bleeding. Results: Japanese patients (N=152) and physicians (N=164) showed different preferences. In particular, among non-fatal outcomes, patients considered disabling stroke to be 16 times more important than non-major clinically relevant bleeding and 2.6 times more important than extra-cranial major bleeding. In contrast, physicians considered the same stroke risk to be 2.7 times more important than non-major clinically relevant bleeding and equally important as major bleeding. Conclusions: Results suggest that Japanese patients are willing to tolerate a greater risk of bleeding in exchange for stroke prevention than are Japanese physicians. The findings demonstrate the importance of physician-patient communication in treatment decisions involving stroke preventative therapies.


Development, validation, and clinical implementation of an assay to measure total antibody response to naglazyme (galsulfase).
Naglazyme (galsulfase, rhASB) was developed as enzyme replacement therapy for mucopolysaccharidosis type VI. Naglazyme generated an IgG antibody response in most patients. To better characterize Naglazyme immunogenicity, a solution phase bridged immunoassay was developed to measure total antibody response regardless of isotype. Overnight incubation of serum dilutions with rhASB labeled with biotin and ruthenium-based tags allowed antibody-antigen complexes to form prior to capture on a streptavidin plate. Neat serum was tolerated in the assay, with a 1:10 screening dilution implemented for testing. At this dilution, the assay was sensitive to 75 ng/ml anti-rhASB. Titers were reported as the highest dilution factor with signal above a 95% confidence interval from naïve individual sera. Precise measurement of titers, within two consecutive dilution factors, was observed across analysts and days. Clinical samples showed similar positive/negative results between the IgG ELISA and the total antibody ECLA, although with an imperfect correlation. Improvements in assay performance and implementation strategy altered some positive clinical samples to negative and vice versa. Comparison of the titer readout for clinical samples with the screening signal illustrates a range of relationships for signal versus sample dilution factor, confirming that signal from a screening dilution cannot directly predict the reported titer.


A novel replicon occurring naturally in Escherichia coli is a phage-plasmid hybrid.
A novel DNA replicon in Escherichia coli was identified. It is the smallest natural isolate (1282 bp) found so far. In the presence of phage M13 it grows as a filamentous single-stranded DNA phage. Contrary to previously identified mini-phages this replicon displays sequence homology only to parts of the M13 viral and complementary strand origin. In the absence of M13 this DNA replicates autonomously. The only gene (arp) of the replicon encodes a 32-kd protein, which is essential for autonomous replication. The host rep gene required for replication of single-stranded DNA phages is dispensable. Distinct replication mechanisms are thus involved during growth as defective phage or as autonomous plasmid.


Development of the pharyngeal arch skeleton in Catostomus commersonii (Teleostei: Cypriniformes).
Skeletal elements of the gill arches of adult cypriniform fishes vary widely in number, size, and shape and are important characters in morphologically based phylogenetic studies. Understanding the developmental basis for this variation is thus phylogenetically significant but also important in relation to the many developmental genetic and molecularly based studies of the early developing and hence experimentally tractable gill arches in the zebrafish, a cyprinid cypriniform. We describe the sequence of the chondrification and ossification of the pharyngeal arches and associated dermal bones from Catostomus commersonii (Catostomidae, Cypriniformes) and make selected comparisons to other similarly described pharyngeal arches. We noted shared spatial trends in arch development including the formation of ventral cartilages before dorsal and anterior cartilages before posterior. Qualitatively variable gill arch elements in Cypriniformes including pharyngobranchial 1, pharyngobranchial 4, and the sublingual are the last such elements to chondrify in C. commersonii. We show that the sublingual bone in C. commersonii has two cartilaginous precursors that fuse and ossify to form the single bone in adults. This indicates homology of the sublingual in catostomids to the two sublingual bones in the adults of cobitids and balitorids. Intriguing patterns of fusion and segmentation of the cartilages in the pharyngeal arches were discovered. These include the individuation of the basihyal and anterior copula through segmentation of a single cartilage rod, fusion of cartilaginous basibranchials 4 and 5, and fusion of hypobranchial 4 with ceratobranchial 4. Such "fluidity" in cartilage patterning may be widespread in fishes and requires further comparative developmental studies.


Baroreflex sensitivity and the blood pressure response to beta-blockade.
The objective of this analysis was to determine whether changes in baroreflex sensitivity (BRS) within 35 hypertensive patients (25 M, 10 F, mean age 47 years) treated with beta-blockade as monotherapy relate to reductions in ambulatory blood pressure (BP) or its variability. BP was recorded intra-arterially directly from the brachial artery before and during submaximal exercise. BRS was determined by the phenylephrine injection technique. MAP and its variability were determined for the awake period of 24-h BP monitoring. Subjects were randomised to one of atenolol, metoprolol, pindolol, or propranolol, and restudied after a mean of 5 months. Beta-blockade increased BRS in 24 patients and decreased BRS in 11. BRS increased from 6.53+/-4.94 to 9.40+/-8.62 ms/mm Hg (mean +/- s.d.) (P<0.01). Waking ambulatory MAP decreased from 125.8+/-15.8 to 106.4+/-16.2 mm Hg (P<0.0001), but its variability did not change. Higher BRS after chronic beta-blockade was associated with a decrease in waking ambulatory MAP (r = -0.55, P<0.001), but not with its variability (r = -0.08). Beta-blockade attenuated the pressor response to exercise, but there was a positive relationship between the effect of beta-blockade on BRS, and on the rise in systolic BP during bicycling (r = 0.63; P<0.001). Any dampening effect of beta-blockade on BP variability at rest in hypertensive patients with the greatest increase in BRS may be offset by increased pressor responses to physical activity such as exercise. Consequently, BP variability is unaffected, even though reductions in ambulatory BP during chronic beta-blockade are inversely related to changes in BRS. BP responses to beta-blockade may be a function of the action of this class of drugs on BRS. However, there is considerable variation, between subjects, in their effect on BRS. This may have implications for other conditions, such as dilated cardiomyopathy, or following myocardial infarction, in which improvement in BRS is one mechanism by which beta-adrenoceptor blockade could improve survival.