Clinical mutational profiling of bone metastases of lung and colon carcinoma and malignant melanoma using next-generation sequencing.
Bone is a common metastatic site for solid tumors and is often the only source for molecular testing. Current routine decalcification protocols for the processing of bone specimens damage nucleic acids, leading to a high failure rate. In this retrospective quality-assessment analysis, preanalytic factors that contributed to the failure of mutational profiling in metastatic bone specimens were evaluated using a next-generation sequencing assay. Mutational profiling was conducted in 33 formalin-fixed, paraffin-embedded bone lesions that were submitted to a clinical laboratory. Adequate depth of coverage was obtained in 21 specimens, of which, 16 had mutations detected. "No results" were reported in 12 specimens because the NGS assay failed. There was a significantly higher failure rate in bone specimens compared with nonbone specimens (36% vs 2.3%, respectively). Although nonbone specimens had a higher failure rate in biopsy/fine-needle aspiration (FNA) specimens, in-house bone biopsy/FNA specimens with or without short-duration surface decalcification had a lower failure rate than resected bone specimens (11% vs 60%, respectively). The high failure rate in resected metastatic bone specimens was associated with regular decalcification but not with low DNA input. Next-generation sequencing assays demonstrated clinical utility in metastatic bone specimens. FNA (smear and cell block) and core-biopsy specimens provided adequate resources of nucleic acids for molecular profiling of metastatic bone lesions. The current findings suggest the need for developing specific tissue procurement and processing protocols for bone metastases and greater use of small biopsy and FNA specimens. Cancer Cytopathol 2016;124:744-53. © 2016 American Cancer Society.


Association between level of education and oral health status in 35-, 50-, 65- and 75-year-olds.
The aim of the present study was to evaluate the association between educational level and dental disease, treatment needs and oral hygiene habits. Randomized samples of 35-, 50-, 65- and 75-year-olds, classified according to the educational level: [low (LE): elementary school or higher (HE)], were identified. In 1091 subjects, a number of characteristics such as (i) number of teeth, (ii) periodontal attachment levels (PAL), (iii) caries and (iv) occlusal function were recorded. Educational level, oral hygiene and dietary habits were self-reported. Non-parametric variables were analyzed by chi2, Mann-Whitney U-Wilcoxon's rank sum tests, and parametric variables by Student's t-test (level of significance 95%). A two-way anova was performed on decayed, missing and filled surfaces to investigate the interaction between age and educational level. All statistical procedures were performed in the SPSS statistical package. The number of remaining teeth was similar for LE and HE in the 35-year olds (25.8 versus 26.6), but in the older age groups LE had significantly a larger number of missing teeth. The LE groups (except in 65-year olds) exhibited significantly more PAL loss. LE had significantly fewer healthy gingival units in all but the 75-year age group. In all age groups, LE had fewer intact tooth surfaces and a significantly poorer occlusal function. The frequency of tooth cleaning measures and dietary habits did not differ between LE and HE. Educational level was shown to influence the oral conditions and should be considered in assessing risk, and in planning appropriate preventive measures.


K+ channel currents in rat ventral prostate epithelial cells.
Electrophysiological function of the normal prostate has not been extensively studied. In particular, ion channel currents and their regulation have not been studied in freshly-isolated prostate cells. Rat prostate secretory epithelial (RPSE) cells were isolated by collagenase treatment. Columnar epithelial cells were used for nystatin-perforated, whole-cell voltage clamp, and the intracellular Ca(2+) concentration ([Ca(2+)](i)) was measured using fura-2. Step-like depolarizing pulses (900 msec) starting from - 90 mV induced outwardly rectifying K(+) currents without inactivation. ACh (10 microM) or ATP (100 microM) increased the outward current and hyperpolarized the cell membrane potential. Ionomycin (0.1 microM), a Ca(2+) ionophore, induced a similar increase in the outward current. TEA (5 mM), charybdotoxin (50 nM), and iberiotoxin (30 nM) inhibited the effect of ACh (or ATP) on the outward current, whereas apamin (100 nM) had no effect. The [Ca(2+)](i) of RPSE cells was increased by ACh, ATP, and UTP. RPSE cells have iberiotoxin-sensitive Ca(2+)-activated K(+) channels that may play an important role in the exocrine secretions of the prostate.


Understanding the effect of residual aluminum salt coagulant on activated sludge in sequencing batch reactor: Performance response, activity restoration and microbial community evolution.
To investigate the effect of residual coagulant after coagulation pretreatment on activated sludge system of wastewater treatment plants (WWTPs), comparative evaluation of lab-scale sequencing batch reactors under different poly-aluminum chloride (PAC) concentrations (20 and 55 mg/L), presenting the performance differences of reactors. Results showed that the PAC concentration of 20 mg/L slightly enhanced the average removal efficiencies of chemical oxygen demand (COD) and total nitrogen (TN), up to 93.43% and 72.52%. Whereas, an inhibition effect was exerted at the PAC concentration of 55 mg/L, the average removal efficiencies decreased to 88.56% and 57.80% respectively. Similarly, the residual aluminum salts showed a concentration effect of low promotion and high inhibition on sludge activity index. The content of specific oxygen utilization rate (SOUR) and dehydrogenase (DHA) sharply decreased by 30.17% and 53.56% under the high PAC concentration of 55 mg/L. Activity recovery phase showed that the suppression of aluminum salt coagulant on biological system was reversible. High-throughput sequencing presented that the relative abundance of microbes showed obvious variations at different PAC concentrations, and certain bacteria in Chloroflexi and Bacteroidota exhibited better adaptability to the high PAC concentration environment. Nevertheless, the antagonism action between denitrifying genera and other genera as well as the downregulation of functional enzymes regarding nitrogen metabolism gave rise to the deterioration of denitrification under the high PAC concentration of 55 mg/L. This study revealed the influence mechanism of residual aluminum salt coagulant on activated sludge system, providing strategies for efficient decontamination and long-term stable operation of biological system in wastewater treatment plant under the condition of adding PAC.


Identification and Characterization of Some Genes, Enzymes, and Metabolic Intermediates Belonging to the Bile Acid Aerobic Catabolic Pathway from Pseudomonas.
The study of the catabolic potential of microbial species isolated from different habitats has allowed the identification and characterization of bacteria able to assimilate bile acids and/or other steroids (e.g., testosterone and 4-androsten-3,17-dione) under aerobic conditions through the 9,10-seco pathway. From soil samples, we have isolated several strains belonging to genus Pseudomonas that grow efficiently in chemically defined media containing some cyclopentane-perhydrophenanthrene derivatives as carbon sources. Genetic and biochemical studies performed with one of these bacteria (P. putida DOC21) allowed the identification of the genes and enzymes belonging to the route involved in bile acids and androgens, the 9,10-seco pathway in this bacterium. In this manuscript, we describe the most relevant methods used in our lab for the identification of the chromosomal location and nucleotide sequence of the catabolic genes (or gene clusters) encoding the enzymes of this pathway, and the tools useful to establish the role of some of the enzymes that participate in this route.


Dopamine D3 receptor in the nucleus accumbens alleviates neuroinflammation in a mouse model of depressive-like behavior.
We recently reported that dopamine D3 receptor (D3R) was involved in inflammation-related depression. Nucleus accumbens (NAc) inflammation is implicated in the development and progression of depression, but its regulatory mechanism remains largely unknown. In a mouse model of NAc neuroinflammation induced by bilateral NAc injection of lipopolysaccharide (LPS), we observed that NAc neuroinflammation triggered depressive-like behaviors, and D3R expression decline and microglial activation in the NAc. A selective knockdown of D3R in the NAc elicited depressive-like behaviors, while re-expression of D3R in the NAc of global D3RKO mice alleviated depressive-like behaviors induced by D3R deficiency. D3R downregulation in the NAc shifted microglia toward a proinflammatory state, which was validated with cultured mouse microglial cultures. Further in vitro results demonstrated that D3R inhibition induced microglia to enter a proinflammatory state primarily through the Akt signaling pathway. In conclusion, our results suggest that D3R expression in the NAc may inhibit microglial proinflammatory responses in the NAc, thus alleviating NAc neuroinflammation and subsequent depressive-like behaviors through the Akt signaling pathway.


Acid and neutral lipases of cultured adipocytes of infants and children.
In order to determine whether the mobilization of intracellular triglycerides observed in cultured human adipocytes is associated with changes in the activities of acid and neutral lipases, the activities of both enzymes were measured weekly on cultured adipocytes for several months. The activity of acid lipase was initially very low, but rose to levels 40 times the original activity within 3 months. The activity of neutral lipase decreased rapidly within the first 2-4 weeks and remained at approximately 25% of original levels thereafter.


Magnitude and categories of pedestrian fatalities in South Africa.
Road traffic injuries, a major global public health burden, are concentrated in low-income and middle-income countries.(1) In contrast to high-income countries, pedestrians make up the largest group of road traffic injuries and fatalities in low- and middle-income countries. This article presents an analysis of pedestrian fatalities (2001-2004) in four South African cities. The article describes the magnitude, demographic, and temporal factors associated with pedestrian fatalities and presents a typological analysis to identify particular groups of at risk pedestrians. The analysis can serve to inform the development of prevention programmes tailored to the needs of specific at risk pedestrian groups. Data were obtained from the National Injury Mortality Surveillance System (NIMSS). The results indicated that there were a total of 7433 pedestrian deaths (2001-2004) for the four cities and the majority occurred over weekends. Most (56.7%) were between ages 20 and 44 years. Overall, there were 3.3 male pedestrian deaths for every female pedestrian death, and over half (58%) of the 4004 cases tested were positive for alcohol. A typological analysis identified three categories of pedestrian fatalities: (1) male pedestrian fatalities that showed high levels of alcohol concentrations, (2) female and elderly pedestrian deaths that occurred between 6 AM and midday, and (3) children, adolescents, and young adult pedestrian fatalities that typically occurred during weekday afternoons and evenings. The findings call for multiple strategies for combating pedestrian fatalities.


Ultrahydrophobicity indicates a non-adhesive default state in gecko setae.
Geckos may represent the world's most demanding adhesives application. The adhesive setae on the toes of climbing geckos must adhere strongly yet avoid fouling or attachment at inappropriate times. We tested the hypothesis that gecko setae are non-adhesive in their unloaded default state by comparing the water droplet contact angle (theta) of isolated setal arrays to the smooth surface of eye spectacle scales of tokay geckos (Gekko gecko). At equilibrium, theta was 98.3 +/- 3.4 degrees in spectacle scales of live geckos and 93.3 +/- 3.5 degrees in isolated spectacles. Isolated setal arrays were ultrahydrophobic, with theta of 160.6 +/- 1.3 degrees (means +/- SD). The difference in theta of setal arrays and smooth spectacles indicates a very low contact fraction. Using Cassie's law of surface wettability, we infer that less than 6.6% of the surface of unloaded setae is solid and at least 93.4% is air space. We calculated that the contact fraction must increase from 6.6% in the unloaded state to 46% in the loaded state to account for previously measured values of adhesion. Thus gecko setae may be non-sticky by default because only a very small contact fraction is possible without mechanically deforming the setal array.


Is the nasal cycle an artifact? The role of asymmetrical postures.
Both the nasal cycle and postural asymmetries between the lateral halves of the body will cause the patent cavity of the nasal airway to change sides. Therefore, the cycle might result from adjustments of posture during prolonged periods of nasal resistance measurements. Posterior rhinometric measurements of unilateral nasal resistance were made three times for 5 to 7 hours on a patient in whom 11 periods of lateral recumbency had not induced nasal resistance changes. A nasal cycle was observed. It is concluded that the cycle is not the product of asymmetrical body pressures.


Synthesis and biological activity of polyprenols.
The polyprenols and their derivatives are highlighted in this study. These lipid linear polymers of isoprenoid residues are widespread in nature from bacteria to human cells. This review primarily presents the synthesis and biological activities of polyprenyl derivatives. Attention is focused on the synthesis and biological activity of dolichols, polyprenyl ester derivatives and polyprenyl amines. Other polyprenyl derivatives, such as oxides of polyprenols, aromatic polyprenols, polyprenyl bromide and polyprenyl sulphates, are mentioned. It is noted that polyprenyl phosphates and polyprenyl-linked glycosylation have better antibacterial, gene therapy and immunomodulating performance, whereas polyprenyl amines have better for antibacterial and antithrombotic activity. Dolichols, polyprenyl acetic esters, polyprenyl phosphates and polyprenyl-linked glycosylation have pharmacological anti-tumour effects. Finally, the postulated prospect of polyprenols and their derivatives are discussed. Further in vivo studies on the above derivatives are needed. The compatibility of polyprenols and their derivatives with other drugs should be studied, and new preparations of polyprenyl derivatives, such as hydrogel glue and release-controlled drugs, are suggested for future research and development.


How to classify Raynaud's phenomenon. Long-term follow-up study of 73 cases.
Raynaud's phenomenon without an underlying cause was diagnosed in 96 consecutive patients in 1978 to 1979. Seventy-three patients were available for long-term follow-up. They were classified on initial evaluation as having primary Raynaud's phenomenon (49 patients) when no clinical, laboratory, or serologic abnormality was detected, and as having suspected secondary Raynaud's phenomenon when at least one finding was abnormal. Re-evaluation was performed in 1984 to 1985 after an average duration of Raynaud's phenomenon of 14.9 +/- 12 years. The average duration of the follow-up from initial to final evaluation was 4.7 +/- 1 years. On final evaluation, none of the 49 patients with an initial diagnosis of primary Raynaud's phenomenon had evidence of secondary Raynaud's phenomenon, whereas 14 of the 24 patients with suspected secondary Raynaud's phenomenon had a definite diagnosis. Among them, there were 13 connective tissue diseases. The study proved that Raynaud's phenomenon without an underlying cause must be followed up for more than two years, contrary to what was recommended previously, before it can be rightly diagnosed as primary Raynaud's phenomenon. Moreover, the results suggested that, in order to distinguish early primary Raynaud's phenomenon from suspected secondary Raynaud's phenomenon, a simple and noninvasive evaluation is sufficient. In this study, the evaluation included history and clinical examination, tests for antinuclear antibodies, radiography of hands, chest roentgenography, and nailfold capillary microscopy.


Effects of dibutyryl guanosine 3',5'-cyclic monophosphate and sodium nitroprusside in pepsinogen secretion from guinea pig chief cells with respect to intracellular Ca2+.
Both Ca2+ and adenosine 3',5'-cyclic monophosphate act as intracellular second messengers in pepsinogen secretion from chief cells. However, the role of intracellular guanosine 3',5'-cyclic monophosphate (cGMP) in this process has not been defined. Although dibutyryl cGMP (dbcGMP), a membrane-permeable derivative of cGMP, has been shown to inhibit pepsinogen secretion only stimulated by cholecystokinin (CCK), the intracellular mechanism of this effect remains unclear. We evaluated the role of intracellular cGMP in pepsinogen secretion from monolayer cultured guinea pig chief cells using dbcGMP and sodium nitroprusside, both of which increase intracellular cGMP. Dibutyryl cGMP and sodium nitroprusside have now been shown to inhibit pepsinogen secretion induced by not only CCK octapeptide but also carbamylcholine chloride and ionomycin in a dose-dependent manner. Furthermore, dbcGMP reduced the increase in intracellular free Ca2+ concentration induced by carbamylcholine chloride, CCK octapeptide, and ionomycin. These results suggest that intracellular cGMP may inhibit pepsinogen secretion by reducing the intracellular free Ca2+ concentration.


Long-term follow-up of external dacryocystorhinostomy and the factors affecting its success.
To evaluate the long-term follow-up results of external dacryocystorhinostomy (Ex-DCR) and to assess the role of some factors in its success. Between January 1990 and November 2002, 437 nasolacrimal systems of 387 patients were operated on and included in the study. The success rates were determined on the basis of the last examinations of the patients, and the correlations between success rates and factors such as age, gender, the duration of obstruction, history of acute dacryocystitis, the side of obstruction, and the postoperative follow-up period were calculated. The mean follow-up period was 5.9 +/- 2.5 years. Fifty cases (12.9%) were operated bilaterally. The initial success rate was 91.5% and the last success rate was 98.4% after reoperations. We found meaningful correlations between success and duration of obstruction, age of the patient, and the duration of postoperative follow-up (r = 0.18, 0.17 and -0.14, p = 0.036, 0.001 and 0.002, respectively). In comparison to newly developed surgical techniques for the treatment of nasolacrimal duct obstruction, Ex-DCR still appears to offer higher success rates, even after long-term follow-up. In addition to being a cost-effective procedure, it requires no sophisticated instruments. In our opinion, the high success rates may be explained by the nature of the operation, which creates a fistula between the sac and the nasal mucosa.


Development and validation of a prognostic model for leflunomide discontinuation with abnormal blood tests during long-term treatment: cohort study using data from the Clinical Practice Research Datalink Gold and Aurum.
To develop and validate a prognostic model for LEF discontinuation with abnormal blood test results. Data from the Clinical Practice Research Datalink Gold and Aurum were used for model development and external validation, respectively. Participants prescribed LEF between 1 January 2007 and 31 December 2019 were followed up from 6 months after the first general practitioner prescription to the earliest of date of outcome, death, 5 year follow-up or 31 December 2019. Candidate prognostic factors were ascertained using theory and data-driven approaches. Penalized Cox regression was performed to develop the risk equation, followed by internal validation using 500 bootstraps to correct for optimism. Multiple imputation was applied to handle missing data. Model performance was assessed in terms of calibration and discrimination. Data for 1487 and 2329 participants contributing 3140 and 5246 person-years follow-up were included in the development and validation cohorts, respectively. Thirteen candidate predictors were included in the model. Epilepsy and either cytopenia or elevated liver enzymes during the first 6 months of shared-care LEF prescription were strong predictors of drug discontinuation with a hazard ratio of 4.39 (95% CI 1.74, 11.06) and 3.06 (2.15, 4.35), respectively. The unadjusted and optimism-adjusted calibration slope in development data was 1.00 (95% CI 0.75, 1.25) and 0.72 (95% CI 0.47, 0.97), respectively. The calibration slope in validation data was 0.91 (95% CI 0.74, 1.07). The model showed prognostic separation with an optimism-adjusted Royston D statistic of 0.73 (95% CI 0.44, 1.02). We have developed and externally validated an easy-to-use prognostic model that may be used to risk stratify monitoring for LEF toxicity and to make informed choices about risks when choosing treatments.


Cadmium and mercury nephrotoxicity.
Despite increasing attempts to control environmental pollution, changes in the distribution and bioavailability of toxic metals like mercury and cadmium are still occurring. Apart from natural processes, other contributory factors include the gradual spread of industrialization, the use of sewage sludge as a fertilizer and the acidification of Northern Hemisphere groundwater. Animals (including man and domestic varieties) can accumulate harmful concentrations of toxic metals. We therefore looked for damage to the kidneys in seabirds contaminated with mercury and cadmium and made comparisons with kidneys from three other groups of animals: seabirds from an uncontaminated colony, metal-dosed birds and metal-dosed mice. We report here that, comparing all these groups of animals, individuals with comparatively high levels of metals had nephrotoxic lesions of a similar type and severity. Moreover, the metal concentrations at which damage began and at which biochemical changes could be detected were below those presently considered as relatively safe for humans by the World Health Organization.


Shedding light on stellate cells.
The relationship between grid cells and two types of neurons found in the medial entorhinal cortex has been clarified.


Bioenergetics in the aging Fischer 344 rat: effects of exercise and food restriction.
The capacity for energy production was evaluated in male, Fischer 344 rats as they advanced from adulthood through senescence. At 10 months of age, the animals were divided into three groups: sedentary, fed ad libitum (S); exercised by treadmill running, fed ad libitum (E); and sedentary, caloric restricted by alternate day feeding (R). Activities of selected enzymes, ADP-stimulated respiration and levels of cytochromes, were determined in homogenates of liver and gastrocnemius muscle prepared from young controls (10-month old S) and 18-, 24-, and 30-month old animals. In liver, age-linked decrements were found in the activities of 3-hydroxyacyl-CoA dehydrogenase (S, E, and R) and citrate synthase (S), and in cytochrome c content (S and E), whereas substrate-catalysed oxidations were unaffected. In the gastrocnemius muscle (S, E, and R), respiration, activities of enzymes of the Krebs cycle and glycolysis, and cytochrome content were decreased after the age of 18 months. Oxidative capacity was increased in muscle through exercise (about 40%) and in liver by food restriction (about 20%). Body and soleus muscle mass declined similarly in all groups (about 14% from 30 to 18 months of age), whereas the loss of weight in the gastrocnemius muscle was much greater (34%). The data indicate that energy metabolism in the senescent animal is competent to meet its needs and age-related declines in energy metabolism are secondary to the aging process.


The influence of cognitive reserve on seizure-induced injury.
Status epilepticus (SE) is associated with a significant risk of cognitive impairment. While many factors likely determine cognitive outcome following SE, there is evidence that cognitive ability prior to a neurological insult may be an important determinant of outcome. Patients with greater cognitive abilities or so-called cognitive reserve may be less vulnerable to injury than patients with limited cognitive ability. Here we tested the hypothesis that cognitive abilities prior to SE would be predictive of cognitive outcome. Immature rats were tested in the water maze, a test of visual-spatial memory, and divided into fast and slow learners. Animals were then subjected to SE and retested in the water maze 23 days later. Control rats were tested in the same manner but not subjected to SE. SE resulted in marked impairment in water maze performance. However, no statistical difference was noted in performance between slow and fast learners in either the SE or control group. Likewise, no differences were seen in the histopathology of the slow and fast learners. This study demonstrates that SE adversely effects visual-spatial memory equally in both fast and slow learners and does not support the theory that cerebral reserve plays a major role in cognitive function following a cerebral insult.


Association Between Dose and Duration of Cisplatin Exposure with Cytotoxicity Effect on Nasopharyngeal Carcinoma Stem Cell.
Nasopharyngeal carcinoma (NPC) is ranked 6th of malignant tumors in Indonesia. To analyze the correlation of dose and duration of cisplatin exposure with cytotoxic effects on nasopharyngeal carcinoma stem cells. The biopsy NPC tissue was cultured and processed to obtain NPC stem cells to be treated with cisplatin different doses and durations (24 and 48 h). The number of dead cells after exposure will be calculated using a hemocytometer. Death stem cell density of NPC at exposure of 2 μg/ml cisplatin dose was 81.37%, while the smallest death cell density a dose of 0.05 μg/ml after a 72-h observation was 21.3%. The coefficient correlation 0.827 and value p = 0.000 (p < 0.05). The analysis of the correlation between cisplatin exposure duration and death cell was also significant with the correlation coefficient - 0.357 and the value p = 0.001 (p < 0.05). There was a correlation between the increased dose of cisplatin with the cytotoxicity effects on NPC stem cell.


Estrogen receptor subtypes localization shifts in cultured mouse ovarian follicles.
Estrogens play an important role in the growth, differentiation, and function of female reproductive tissues. Estrogen signals through estrogen receptors (ERs), members of the nuclear receptor superfamily. The two major forms, ERalpha and ERbeta, are expressed in the mouse ovary, where ERbeta is predominantly expressed in granulosa cells, and ERalpha in theca cells. In this study, we determined the expression pattern of ER subtypes within mouse follicles cultured from the early preantral stage up to the preovulatory stage and after an ovulatory stimulus in different culture conditions. Immunohistochemical studies performed at different time points of culture revealed that ERbeta was found exclusively in granulosa cell nuclei regardless of follicular growth stage or culture conditions. In contrast, ERalpha was found in oocyte, granulose, and theca cells, and its subcellular localization differed between follicular growth stages and culture conditions. A shift from a predominant cytoplasmic to a predominant nuclear immunolocalization was observed in granulosa cells as follicles reached the antral growth phase, and was postponed in culture conditions with minimal growth factor supplementation. In response to hCG, ERbeta protein levels in luteinized granulosa cells spectacularly declined to undetectable levels, while ERalpha immunostaining again shifted to cytoplasmic regions, but not in theca cells.


Functional characterization of developmentally critical genes in the white-backed planthopper: Efficacy of nanoparticle-based dsRNA sprays for pest control.
Epidermal growth factor receptor (EGFR), zinc finger homeodomain-2 (zfh-2), Abdominal-A (Abd-A), and Abdominal-B (Abd-B) regulate the growth and development of the insect abdomen. However, their potential roles in pest control have not been fully assessed. The development of insecticide resistance to multiple chemistries in the white-backed planthopper (WBPH), a major pest of rice, has prompted interest in novel pest control approaches that are ecologically friendly. Although pest management approaches based on double-stranded RNA (dsRNA)-mediated RNA interference (RNAi) have potential, their susceptibility to degradation limits large-scale field applications. These limitations, however, can be overcome with nanoparticle-dsRNA complexes that have greater environmental stability and improved cellular uptake. In this study, at 5 days post-injection, transcripts for the four gene targets were reduced relative to controls and all of the experimental groups exhibited significant phenotypic defects and increased mortality. To evaluate the potential of these gene targets for field applications, a nanocarrier-dsRNA spray delivery system was assessed for RNAi efficacy. At 11 days post-spray, significant phenotypic defects and increased mortality were observed in all experimental groups. Taken together, the results confirm the suitability of the target genes (SfEGFR, Sfzfh-2, SfAbd-A, and SfAbd-B) for pest management and demonstrate the efficacy of the nanocarrier spray system for inducing RNAi-mediated knockdown. As such, the study lays the foundation for the further development and optimization of this technology for large-scale field applications. © 2022 Society of Chemical Industry.


Plastomes of Sonchus (Asteraceae) endemic to the Atlantic Madeira archipelago: Genome structure, comparative analysis, and phylogenetic relationships.
The woody Sonchus alliance, a spectacular example of adaptive radiation with six genera and approximately 31 species, is found exclusively on three Macaronesian Islands (Madeira, Canaries, and Cape Verdes) in the Atlantic Ocean. Four of the Sonchus taxa are restricted to Madeira, including shrubs and small trees at higher elevations (S. fruticosus and S. pinnatus), and caudex perennials in the lower coastal areas (S. ustulatus subsp. maderensis and S. ustulatus subsp. ustulatus). The Madeiran Sonchus stemmed from a single colonization event that originated from the Canaries < 3 million years ago. However, the plastome evolution and species relationships remains insufficiently explored. We therefore assembled and characterized the plastomes of four Sonchus taxa from Madeira and conducted a phylogenomic analysis. We found highly conserved plastome sequences among the taxa, further supporting a single and recent origin. We also found highly conserved plastomes among the cosmopolitan weedy Sonchus, Macaronesian Sonchus in the Atlantic, and Juan Fernández Islands Dendroseris in the Pacific. Furthermore, we identified four mutation hotspot regions (trnK-rps16, petN-psbM, ndhF-Ψycf1, and ycf1) and simple sequence repeat motifs. This study strongly supports the monophyly of Madeiran Sonchus. However, its relationship with the remaining woody Sonchus alliance from the Canary Islands requires further investigation.


[Microarray CGH: principle and use for constitutional disorders].
Chips technology has allowed to miniaturize process making possible to realize in one step and using the same device a lot of chemical reactions. The application of this technology to molecular cytogenetics resulted in the development of comparative genomic hybridization (CGH) on microarrays technique. Using this technique it is possible to detect very small genetic imbalances anywhere in the genome. Its usefulness has been well documented in cancer and more recently in constitutional disorders. In particular it has been used to detect interstitial and subtelomeric submicroscopic imbalances, to characterize their size at the molecular level or to define the breakpoints of translocation. The challenge today is to transfer this technology in laboratory medicine. Nevertheless this technology remains expensive and the existence of numerous sequence polymorphisms makes its interpretation difficult. Finally its is unlikely that it will make karyotyping obsolete as it does not allow to detect balanced rearrangements which after meiotic segregation might result in genome imbalance in the progeny.


Astrocyte reactivity in the glia limitans superficialis of the rat medial prefrontal cortex following sciatic nerve injury.
The glia limitans superficialis (GLS) on the rodent cortical surface consists of astrocyte bodies intermingled with their cytoplasmic processes. Many studies have observed astrocyte reactivity in the medial prefrontal cortex (mPFC) parenchyma induced by a peripheral nerve injury, while the response of GLS astrocytes is still not fully understood. The aim of our study was to identify the reactivity of rat GLS astrocytes in response to sciatic nerve compression (SNC) over different time periods. The alteration of GLS astrocyte reactivity was monitored using immunofluorescence (IF) intensities of glial fibrillary acidic protein (GFAP), glutamine synthetase (GS), and NFκBp65. Our results demonstrated that SNC induced GLS astrocyte reactivity seen as increased intensities of GFAP-IF, and longer extensions of cytoplasmic processes into lamina I. First significant increase of GFAP-IF was observed on post-operation day 7 (POD7) after SNC with further increases on POD14 and POD21. In contrast, dynamic alteration of the extension of cytoplasmic processes into lamina I was detected as early as POD1 and continued throughout the monitored survival periods of both sham and SNC operations. The reactivity of GLS astrocytes was not associated with their proliferation. In addition, GLS astrocytes also displayed a significant decrease in GS immunofluorescence (GS-IF) and NFκB immunofluorescence (NFκB-IF) in response to sham and SNC operation compared with naïve control rats. These results suggest that damaged peripheral tissues (following sham operation as well as peripheral nerve lesions) may induce significant changes in GLS astrocyte reactivity. The signaling mechanism from injured peripheral tissue and nerve remains to be elucidated.


The comparison of the survival outcome between robotic-assisted radical prostatectomy and radiation therapy for localized prostate cancer in men over 70 years: Korean Nationwide Observational Study.
This study aimed to investigate the survival after robotic-assisted radical prostatectomy (RARP) in comparison with radiation therapy (RT) in elderly patients (≥ 70 years) with non-metastatic prostate cancer (PCa), given the contemporary Korean life span of 80 years. From the National Health Insurance Sharing Service data, men aged ≥ 70 years diagnosed with PCa from 2006 through 2016 undergoing RARP or RT without systemic chemotherapy or prolonged androgen deprivation therapy (ADT) were identified, then the mortality was compared after adjusting host factors. Data revealed 13,952 patients with RARP (N = 7535) or RT (N = 6417). Patients who underwent RARP were younger (73.81 ± 3.58 vs. 75.05 ± 3.83 years) and had a higher income. While the majority of the RARP group (94.1%) received no additional therapy, 57.4% of the RT group combined ADT. The adjusted mortality rate for total patients was lower in the RARP group (HR = 0.767, 95% CI 0.072-0.818, p < 0.0001). From the age of 75 years, more patients underwent RT (n = 3239) than RARP (n = 2478). When dividing patients by this age cut-off, the adjusted mortality for the younger patient (70-74 years) was lower in the RARP group than the RT group (HR = 0.580, p < 0.0001). In contrast, the adjusted mortality rate for men older than 75 years was similar (HR = 0.98, 95% CI 0.897-1.07, p = 0.649). This study demonstrates that even the patient over 75 years old who could afford to undergo robotic-assisted radical prostatectomy for non-metastatic PCa had a similar survival in comparison with RT, which had long been a standard of care for this particular age group.


Monte Carlo-based noise compensation in coil intensity corrected endorectal MRI.
Prostate cancer is one of the most common forms of cancer found in males making early diagnosis important. Magnetic resonance imaging (MRI) has been useful in visualizing and localizing tumor candidates and with the use of endorectal coils (ERC), the signal-to-noise ratio (SNR) can be improved. The coils introduce intensity inhomogeneities and the surface coil intensity correction built into MRI scanners is used to reduce these inhomogeneities. However, the correction typically performed at the MRI scanner level leads to noise amplification and noise level variations. In this study, we introduce a new Monte Carlo-based noise compensation approach for coil intensity corrected endorectal MRI which allows for effective noise compensation and preservation of details within the prostate. The approach accounts for the ERC SNR profile via a spatially-adaptive noise model for correcting non-stationary noise variations. Such a method is useful particularly for improving the image quality of coil intensity corrected endorectal MRI data performed at the MRI scanner level and when the original raw data is not available. SNR and contrast-to-noise ratio (CNR) analysis in patient experiments demonstrate an average improvement of 11.7 and 11.2 dB respectively over uncorrected endorectal MRI, and provides strong performance when compared to existing approaches. Experimental results using both phantom and patient data showed that ACER provided strong performance in terms of SNR, CNR, edge preservation, subjective scoring when compared to a number of existing approaches. A new noise compensation method was developed for the purpose of improving the quality of coil intensity corrected endorectal MRI data performed at the MRI scanner level. We illustrate that promising noise compensation performance can be achieved for the proposed approach, which is particularly important for processing coil intensity corrected endorectal MRI data performed at the MRI scanner level and when the original raw data is not available.


Synthetic chloride-selective carbon nanotubes examined by using molecular and stochastic dynamics.
Synthetic channels, such as nanotubes, offer the possibility of ion-selective nanoscale pores which can broadly mimic the functions of various biological ion channels, and may one day be used as antimicrobial agents, or for treatment of cystic fibrosis. We have designed a carbon nanotube that is selectively permeable to anions. The virtual nanotubes are constructed from a hexagonal array of carbon atoms (graphene) rolled up to form a tubular structure, with an effective radius of 4.53 Å and length of 34 Å. The pore ends are terminated with polar carbonyl groups. The nanotube thus formed is embedded in a lipid bilayer and a reservoir containing ionic solutions is added at each end of the pore. The conductance properties of these synthetic channels are then examined with molecular and stochastic dynamics simulations. Profiles of the potential of mean force at 0 mM reveal that a cation moving across the pore encounters an insurmountable free energy barrier of ∼25 kT in height. In contrast, for anions, there are two energy wells of ∼12 kT near each end of the tube, separated by a central free energy barrier of 4 kT. The conductance of the pore, with symmetrical 500 mM solutions in the reservoirs, is 72 pS at 100 mV. The current saturates with an increasing ionic concentration, obeying a Michaelis-Menten relationship. The pore is normally occupied by two ions, and the rate-limiting step in conduction is the time taken for the resident ion near the exit gate to move out of the energy well.


Malignant melanoma: sister chromatid exchange analysis in three families.
Sister chromatid exchange (SCE) was analyzed in stimulated lymphocytes and skin fibroblasts in members of three families with cutaneous malignant melanoma (CMM). Two of these families were characterized by familial CMM; the other family had one patient affected by CMM and two others with other cutaneous melanocytic lesions. All the patients had undergone surgery but no chemotherapy. Higher and differing SCE rates were found in lymphocytes and in fibroblasts of all patients. A wide range of SCE distribution was found in patients with high SCE rate. A few healthy close relatives also showed relatively high SCE rates and wide range distributions. These subjects may be regarded as a subset of family members at high risk for developing cancer. The variability of SCE rates and distribution may reflect genetic heterogeneity of CMM.


Symptoms and diagnosis of asthma in a general population – longitudinal results from the SHIP database.
Against the background of an increasing prevalence of allergies, the epidemiology of asthma in relation to age is still a matter of debate. To further clarify the prevalence of asthma, asthmatic symptoms, and their change over time within a population sample, we analyzed longitudinal data from the Study of Health in Pomerania (SHIP). Standardized questionnaires for asthma and asthmatic symptoms were available in 4310 individuals aged 20–79 years [corrected]. The population sample underwent a 5-year follow-up with 3300 subjects reexamined. The questionnaire evaluated estimates to describe the prevalence in wheeze, chest tightness, shortness of breath, coughing, nocturnal symptoms, and asthma attacks in the past 12 months, current medications, current asthma, and nasal allergies at baseline and its longitudinal net change. The prevalence of current asthma at the baseline study was 1.8% [95% confidence interval (CI 1.4–2.2)], those of nasal allergies 16.7% (95% CI 15.6–17.9). Sixteen percent of asthmatic individuals were not medically treated. The net changes in asthmatic symptoms per 60 months of follow-up ranged between a decrease by 2.0% (chest tightness at night) and an increase by 1.2% (nocturnal attack of coughing). The proportions of subjects with current asthma remained unchanged. The prevalence of current asthma and symptoms was higher in young individuals, whereas the net change over time was slightly increasing at the age of 36 and above. Despite the fact of a high prevalence of subjects complaining of nasal allergies, the overall prevalence of current asthma remained low over time. The prevalence in asthma and asthmatic symptoms as well as its net changes over 5 years were age dependent.


Phosphorus-31 magnetic resonance spectroscopy of experimentally induced arthritis in rats.
Phosphorus-31 magnetic resonance spectroscopy (MRS) of the hind paws of rats was performed at 1.5 Tesla before and during the course of an experimentally-induced inflammatory arthritis. Arthritis was induced by daily subcutaneous administration of 6-sulfanilamidoindazole, an antibacterial sulfa known to produce an acute, self-limited arthritis and periarthritis in the hind paws of rats. Phosphorus-31 spectra obtained after the development of clinical arthritis showed a significant (p less than 0.01) increase in the intensity of a group of resonances occurring downfield from phosphocreatine as compared to spectra obtained before treatment. In all rats, this increase correlated well with histological evidence of inflammation as well as with the degree of inflammation judged clinically (r = 0.89, p less than 0.001), and was present before roentgenographic evidence of bony involvement. The use of 31P MRS may permit evaluation of the severity of an inflammatory arthritis with greater accuracy than the bony changes definable by plain roentgenograms.


[Prognosis of the inflammatory complications after radical operations for cancer of the colon and rectum].
The study was concerned with identification of a group at high risk for complications following surgery for colorectal cancer. A correlational analysis of a complicated postoperative clinical course versus patients' initial status and type of surgery was carried out. A regression equation was obtained for predicting immediate results of surgery as well as assessing the effect of the said factors on complication development in the course of correction treatment.


The prevention and conquest of scurvy, beri-beri, and pellagra.
I have described how many years of research were needed to overcome three nutritional diseases that occurred throughout the world. When human beings migrated from tropical and subtropical regions to the temperate zone, they lost their year-round supplies of vitamin C, and scurvy began to occur. The emergence of beri-beri had a mechanical basis; the introduction of milling, which prevented the rancidity of rice and wheat, also removed the essential vitamin B1. The circumstance leading to the third deficiency disease was reliance on maize, without supplementing it with "protective" foods, such as meat, milk, and vegetables. Under conditions of poverty, the protective foods became expensive, and pellagra appeared. Synthetic chemistry, together with improvements in the diet and in education, largely overcame scurvy, beri-beri, and pellagra, but deficiencies of vitamins A, C, and folic acid still occur widely in economically disadvantaged populations, and this is a challenge to those who wish to improve public health. The brilliant prophesies of Casimir Funk, made in 1912, were fulfilled within 25 years. Today, his word "vitamin" is universally familiar. The account of the discovery, identification and synthesis of the vitamins is the story of how human beings have overcome nutritional deficiencies that were imposed by cultural and economic disadvantages or just plain ignorance. These discoveries were a triumph of science and technology. Much of the research depended on the use of experimental animals: guinea-pigs, rats, dogs, monkeys, and chickens. Vitamins are now added to the diets of both animals and human beings.


Effects of morphine and two enzyme resistant enkephalins on schedule-controlled responding in the rat.
Water-deprived rats to respond for access to a water-filled dipper under a 20 response fixed-ratio schedule. After training, the rats were each implanted with a lateral ventricle cannula and the effects of intraventricular injection of morphine, (N-methyl-tyrosyl)1-des COOH-Norleucyl5-Enkephalin, and (D-Ala)2 -(des COOH-Norleucyl)5-Enkephalin were determined. All three compounds produced dose-related decreases in FR20 responding. Morphine was about 5 times more potent than either of the two peptides. Naloxone (1.0 mg/kg, SC) antagonized the response-rate decreasing effects of intraventricular morphine and the two opiate peptides.


Rapid and sensitive detection of Enterobacter sakazakii by cross-priming amplification combined with immuno-blotting analysis.
Enterobacter sakazakii is a widespread and life-threatening bacterium especially in polluted powdered infant milk formula. Several methods have been developed for detection of E. sakazakii such as physiological and biochemical methods, PCR and loop-mediated isothermal amplification. However, these procedures were disadvantages due to a long assay time, low sensitivity or the use of toxic reagents. Our method of cross-priming amplification (CPA) under isothermal conditions combined with immuno-blotting analysis made the whole detection procedure more sensitivity and lower time-consuming. A set of specific displacement primers, cross primers and testing primers were designed based on six specific sequences in E. sakazakii 16S-23S rDNA internal transcribed spacer. Under isothermal condition at 63 °C for 60 min, the specific amplification and hybridization steps were processed simultaneously. The specificity of the CPA was tested in panel of 54 different bacterial strains and 236 milk powder products. Two red signal lines were developed on the BioHelix Express strip in all of positive E. sakazakii strains, and only one signal line was demonstrated by non- E. sakazakii bacterial strains. The limit of decetion of CPA was 6.3 ± 2.7277 fg for the genomic DNA, 88 ± 8.7892 cfu/ml for pure bacterial culture, and 3.2 ± 2.0569 cfu per 100 g milk powder with pre-enrichment. The current study demonstrated that the assay method of CPA combined with immuno-blotting analysis was a specific and sensitive detection for the rapid detection of E. sakazakii.


Systemic hypoxia inhibits T cell response by limiting mitobiogenesis via matrix substrate-level phosphorylation arrest.
Systemic oxygen restriction (SOR) is prevalent in numerous clinical conditions, including chronic obstructive pulmonary disease (COPD), and is associated with increased susceptibility to viral infections. However, the influence of SOR on T cell immunity remains uncharacterized. Here we show the detrimental effect of hypoxia on mitochondrial-biogenesis in activated mouse CD8+ T cells. We find that low oxygen level diminishes CD8+ T cell anti-viral response in vivo. We reveal that respiratory restriction inhibits ATP-dependent matrix processes that are critical for mitochondrial-biogenesis. This respiratory restriction-mediated effect could be rescued by TCA cycle re-stimulation, which yielded increased mitochondrial matrix-localized ATP via substrate-level phosphorylation. Finally, we demonstrate that the hypoxia-arrested CD8+ T cell anti-viral response could be rescued in vivo through brief exposure to atmospheric oxygen pressure. Overall, these findings elucidate the detrimental effect of hypoxia on mitochondrial-biogenesis in activated CD8+ T cells, and suggest a new approach for reducing viral infections in COPD.


Least-squares estimators for the center and radius of circular patterns.
Applying the least-squares methods to data points of a pattern border, we have estimated the center and radius of a circular pattern.


Ultraviolet light immunomodulation of canine islets for prolongation of allograft survival.
Ultraviolet (UV) light treatment of donor islets has been shown to be effective for the prolongation of islet allograft survival in rodent models. This study evaluated UV as an immunomodulator of canine islets. The effects of UV irradiation on islet secretory function in vitro revealed a trend of increasing basal insulin release with increasing doses of UV and a corresponding significant decrease in glucose-mediated insulin release (expressed as percentage of basal fractional insulin release) beginning at UV light exposures of 200-300 J/m2 (n = 3, P less than 0.05). Proliferative responses to UV-irradiated allogeneic peripheral blood leukocytes and islets were significantly decreased by 53-112% (P less than 0.05) in 27 of 29 mixed-lymphocyte cultures and by 35-74% (P less than 0.05) in 4 of 5 mixed-lymphocyte islet culture experiments, respectively, beginning at 200-600 J/m2. Autotransplantation of nonirradiated (n = 8) and irradiated islets (600 J/m2, n = 6) resulted in a 1-mo graft survival rate of 75% for the control group and 50% for the irradiated group. Allotransplantation of irradiated islets (600 J/m2) into either nonimmunosuppressed recipients (1 donor to 1 recipient, n = 8) or recipients of subimmunosuppressive doses of cyclosporin (2 donors to 1 recipient, n = 4) resulted in 100% rejection by day 10. In contrast, when islets were cultured for 24 h postirradiation and transplanted into cyclosporin-treated pancreatectomized recipients (2 donors to 1 recipient), 3 of 7 grafts were prolonged beyond day 10 to days 16, 26, and greater than 100.(ABSTRACT TRUNCATED AT 250 WORDS)


Timing of Tracheostomy for Prolonged Respiratory Wean in Critically Ill Coronavirus Disease 2019 Patients: A Machine Learning Approach.
To propose the optimal timing to consider tracheostomy insertion for weaning of mechanically ventilated patients recovering from coronavirus disease 2019 pneumonia. We investigated the relationship between duration of mechanical ventilation prior to tracheostomy insertion and in-hospital mortality. In addition, we present a machine learning approach to facilitate decision-making. Prospective cohort study. Guy's & St Thomas' Hospital, London, United Kingdom. Consecutive patients admitted with acute respiratory failure secondary to coronavirus disease 2019 requiring mechanical ventilation between March 3, 2020, and May 5, 2020. Baseline characteristics and temporal trends in markers of disease severity were prospectively recorded. Tracheostomy was performed for anticipated prolonged ventilatory wean when levels of respiratory support were favorable. Decision tree was constructed using C4.5 algorithm, and its classification performance has been evaluated by a leave-one-out cross-validation technique. One-hundred seventy-six patients required mechanical ventilation for acute respiratory failure, of which 87 patients (49.4%) underwent tracheostomy. We identified that optimal timing for tracheostomy insertion is between day 13 and day 17. Presence of fibrosis on CT scan (odds ratio, 13.26; 95% CI [3.61-48.91]; p ≤ 0.0001) and Pao2:Fio2 ratio (odds ratio, 0.98; 95% CI [0.95-0.99]; p = 0.008) were independently associated with tracheostomy insertion. Cox multiple regression analysis showed that chronic obstructive pulmonary disease (hazard ratio, 6.56; 95% CI [1.04-41.59]; p = 0.046), ischemic heart disease (hazard ratio, 4.62; 95% CI [1.19-17.87]; p = 0.027), positive end-expiratory pressure (hazard ratio, 1.26; 95% CI [1.02-1.57]; p = 0.034), Pao2:Fio2 ratio (hazard ratio, 0.98; 95% CI [0.97-0.99]; p = 0.003), and C-reactive protein (hazard ratio, 1.01; 95% CI [1-1.01]; p = 0.005) were independent late predictors of in-hospital mortality. We propose that the optimal window for consideration of tracheostomy for ventilatory weaning is between day 13 and 17. Late predictors of mortality may serve as adverse factors when considering tracheostomy, and our decision tree provides a degree of decision support for clinicians.


Fatty fish consumption reduces lipophilic index in erythrocyte membranes and serum phospholipids.
Lipophilic index (LI) has been introduced to assess the overall fatty acid lipophilicity and as a simple estimate of membrane fluidity. However, little is known on effect of diet on LI. We tested if Camelina sativa oil (CSO) high in ALA, fatty fish (FF) or lean fish (LF) affect LI as compared to control diet and, secondarily, if the LI is associated with HDL lipids and functionality and LDL lipidome. We used data from two randomized clinical trials. The AlfaFish intervention lasted 12 weeks and 79 subjects with impaired glucose tolerance were randomized to FF, LF, CSO or control group. In the Fish trial, 33 subjects with myocardial infarction or unstable ischemic heart attack were randomized to FF, LF or control group for 8 weeks. LI was calculated from erythrocyte membrane fatty acids in AlfaFish and from serum phospholipids in Fish trial. HDL lipids were measured using high-throughput proton nuclear magnetic resonance spectroscopy. There was a significant decrease in LI in the FF group in the AlfaFish (fold change 0.98 ± 0.03) and in the Fish trial (0.95 ± 0.04) and the decrease differed from that of control group in both trials and from CSO group in the AlfaFish study. There were no significant changes in LI in LF or CSO groups. The mean diameter of HDL particles and concentration of large HDL particles were inversely associated with LI. FF consumption decreased LI indicating better membrane fluidity in subjects with impaired glucose tolerance or coronary heart disease.


Design of invisibility anti-cloak for two-dimensional arbitrary geometries.
In this paper, we derive the material parameter formulae for designing an electromagnetic invisibility anti-cloak of two-dimensional arbitrary geometry, which is conformal with the cloaked object. Different shapes of electromagnetic invisibility anti-cloaks are proposed to verify the correctness and effectiveness of the proposed formulae. The simulation results show that the invisibility anti-cloak can break cloak shielding and make the external electromagnetic waves into the cloak. This is not only to realize the transfer of information, but will not affect the role of cloak of stealth.


Spirituality, psychotherapy and music in palliative cancer care: research projects in psycho-oncology at an oncology center in Switzerland.
It is recognized as increasingly important in palliative care that spiritual needs of terminally ill patients should be acknowledged and addressed. Two research projects investigated the feasibility of psychotherapeutic and music therapeutic assistance offered to advanced cancer patients. The first project (1998-2000) sought to improve the understanding of the effect of therapeutic support given to 80 patients and the characteristics of the dying process. The second project (2000-2003) assessed the significance of spiritual experiences in illness and affliction. Empathic therapeutic assistance, observations and systematic record keeping were combined with statistical assessment in an interdisciplinary approach. A respectful attitude and spiritual care were taken to perceive and analyse changes in border areas of life. The first project studied the rules and methods of terminal communication and described three stages in the dying process. After a 'passing through,' the dying often had a spiritual opening leading in a state beyond all pain. The second project concentrated on spiritual experiences. Of 251 treated patients, 135 had such experiences. Spiritual experiences can have a great impact on physical and emotional well-being (alleviate pain, release from anxiety and despair, engender feelings of serenity and wholeness) and facilitate dying. Music therapy, psychotherapy and spiritual assistance offer essential methods for psycho-oncology and palliative care. A holistic and interdisciplinary approach is needed to assist patients in their complex suffering. Our findings on spiritual experience and terminal communication should stimulate further research in a still unexplored territory of clinical research.


A general procedure for the purification of human beta-secretase expressed in Escherichia coli.
Expression and purification of human beta-secretase (BACE1) in bacteria have been plagued with issues concerning solubility, inhomogeneous N-terminus, and lack of enzymic activity. Several forms of the mature human BACE1 have been expressed in Escherichia coli with different N-terminal extensions and without the C-terminus transmembrane domain. Although each of the proteins expresses in inclusion bodies, a generalized protocol has been developed to solubilize, refold, and purify these BACE1 variants. The resultant proteins are homogeneous and monodispersed in solution. Each possesses a unique N-terminus. Activity assays using the peptide substrate 7-methoxycoumarin-4-yl-SEVNLDAEFK-2,4-dinitrophenyl-RR, corresponding to the beta-secretase cleavage sequence in the amyloid precursor protein with the Swedish mutations of N(670)L(671) substituting for the residues K(670)M(671), reveal a kcat and KM of 9.3 min(-1) and 55 microM, respectively.


Exploring the therapeutic promise of targeting Rho kinase in rheumatoid arthritis.
Rheumatoid arthritis (RA) is a prevalent systemic autoimmune disease caused by dysregulated inflammatory reactions, T lymphocyte invasion into the joints, and articular thickening. Immune cells, primarily tumor necrosis factor-alpha (TNF-α) and chemokines (interleukin or IL-1), which are predominantly generated by activated macrophages cells, have also been involved with the pathogenesis of rheumatoid arthritis. Rho GTPases are integral factors of biochemical cascades utilized by antigens, and also by cellular receptors, cytokines, and chemokines, to modulate inflammatory reactions, according to growing data. The Rho family is a group of G proteins that govern a variety of biological and physiological activities such as mobility, actin stress fiber production, growth, and polarity. Research suggests that the Rho A and Rho-associated coiled-coil kinase (ROCK) regulatory cascade could be essential in several autoimmune conditions, including RA. ROCK is activated in the synovial of rheumatoid arthritis patients, while the blocking of ROCK with fasudil could also decrease IL-6, TNF-α, and IL-1. This review covers current developments in understanding the overactivation of Rho enzyme activity in RA suppressed by ROCK inhibitors which can be utilized for the treatment of autoimmune disease. We offer an outline of the function of ROCK inhibitors in immune cells and discuss findings which emphasize the rising participation of this category of kinases within the pathological process of autoimmune disorders. Assuming the potential ability of ROCK as a therapeutic, we define approaches that might be used to inhibit Rho kinase activity in rheumatoid disorders.


MiR-21 down-regulation suppresses cell growth, invasion and induces cell apoptosis by targeting FASL, TIMP3, and RECK genes in esophageal carcinoma.
miR-21 is overexpressed in esophageal squamous cell carcinoma (ESCC) and is thought to be correlated with the development of the cancer. The target gene of miR-21 including FASL, TIMP3 and RECK is revealed by researchers. miR-21 may be involved in the tumorgenesis of ESCC by targeting FASL, TIMP3 and RECK. The purpose of this study was to explore the mechanism of miR-21 in the development of ESCC. miR-21 expression in ESCC and the matched non-malignant adjacent tissues (NMATs) was examined by qRT-PCR. Cell growth, cell apoptosis and cell invasion ability of EC9706 and EC-1 cells was examined after the cells were transfected with miR-21 inhibitor. The potential target genes of miR-21 including FASL, TIMP3 and RECK were examined by western blot and Luciferase reporter assay. miR-21 expression was increased significantly in ESCC tissues compared with NMAT. miR-21 down-regulation inhibits cell growth, cell invasion and induces cells to apoptosis. FASL, TIMP3 and RECK are direct targets of miR-21. miR-21 down-regulation inhibits cell growth, invasion and induces cells to apoptosis by targeting FASL, TIMP3 and RECK genes.


Clinical Findings Associated with Ornithodoros brasiliensis Tick Parasitism in Travelers, Southern Brazil.
We report a series of clinical cases associated with parasitism by the Ornithodoros brasiliensis tick in a group of travelers in the Caxias do Sul municipality, Southern Brazil. These cases draw attention to underdiagnosed noninfectious syndromes caused by ticks with restricted local distributions.


Rapid detection of hepatitis B surface antigen by double antibody radioimmunoassay.
HBsAg and anti-HBs can be detected by double-antibody radioimmunoassay in a reliable routine overnight assay of sensitivity approaching that of solid-phase immunoradiometric assay. The test requires careful standardisation and regular preparation of I125-HBsAg of appropriate quality, but otherwise uses only small quantities of commercially available reagents. In this paper, the reaction kinetics of the first incubation (test sample + limiting amount of rabbit anti-HBs), second incubation (reaction mixture + I125 -HBsAg) and precipitation reaction (reaction mixture + antirabbit globulin) were examined, to determine the minimum time required for the assay compatible with adequate sensitivity and user convenience; in addition, in order to reduce the time of radioactive counting, the maximum amount of I125-HBsAg per test compatible with a satisfactory assay was examined. It was concluded that, by terminating each reaction before equilibrium was reached, results could be obtained in 2--3 hours with a detection limit similar to the standard overnight procedure. To allow routine examination of large numbers of samples, the possibilities of complete automation of this liquid-phase type of assay were discussed.


The antinociceptive effect of combined systemic administration of morphine and the glycine/NMDA receptor antagonist, (+)-HA966 in a rat model of peripheral neuropathy.
1. We evaluated the ability of the functional antagonist at the glycine site of the N-methyl-D-aspartate (NMDA) receptor complex, (+)-(1-Hydroxy-3-aminopyrrolodine-2-one) ((+)-HA966), to modulate the antinociceptive action of systemic morphine in a rat model of neuropathic pain produced by chronic constriction injury to the sciatic nerve. Mechanical (vocalization threshold to hindpaw pressure) and thermal (struggle latency to hindpaw immersion into a water bath) stimuli were used. 2. In the mechanical test, morphine (0.05, 0.1 and 0.3 mg kg(-1), i.v.) alone produced dose-dependent effects in both neuropathic and uninjured rats. Likewise, morphine (0.1, 0.3 and 1 mg kg(-1), i.v.) dose-dependently increased struggle latencies of the nerve-injured hindpaw in the hot noxious (46 degrees C) test but was ineffective in the non-noxious warm (44 degrees C) and cold (10 degrees C) test. 3. Pretreatment with (+)-HA966 (2.5 mg kg(-1), s.c.) dose-dependently enhanced the effect of morphine in the mechanical test with the relative potency being nerve-injured hindpaw > contralateral hindpaw > uninjured rat. 4. Likewise, (+)-HA966 dose-dependently enhanced the effect of morphine against a hot (46 degrees C) stimulus and produced, in combination with morphine, a dose-dependent effect against a warm (44 degrees C) stimulus. In the cold (10 degrees C) test, (+)-HA966 reversed the ineffectiveness of the highest dose of morphine. 5. Naloxone blocked the effect of the combination of (+)-HA966 with morphine in all tests. The drug combination produced no motor deficits in animals using the rotarod test. 6. These findings suggest that combined administration of antagonists, acting at the glycine site of the NMDA receptor complex and morphine may be a promising approach in the treatment of neuropathic and acute pain.


Hysterosalpingographic follow-up of the partial salpingectomy type of sterilization.
Two hundred and eleven of 743 patients with salpingectomy-type sterilization had hysterosalpingographic (HSG) follow-up. This type of sterilization was performed post partum, electively suprapubic, vaginally (including fimbriectomies), or at the time of cesarean section. A Foley catheter technique was used for hysterosalpingography. Of these, 1.895 per cent had fistula on hysterosalpingography. Mechanism of fistula formations and recanalization is discussed. A modification of the Pomeroy sterilization procedure is described and discussed.


Human erythrocyte thiopurine methyltransferase: radiochemical microassay and biochemical properties.
A radiochemical micromethod for the determination of thiopurine methyltransferase (TPMT) activity in human red blood cells (RBC) is described. Both 6-mercaptopurine and 6-thioguanine were substrates for the TPMT activity in the human RBC. Apparent Michaelis-Menten (KM) values for 6-mercaptopurine and 6-thioguanine were 3.2 X 10(-4) M and 2.0 X 10(-4) M, respectively. The apparent KM value for S-adenosyl-L-methionine, a co-substrate for the reaction, was 1.7 X 10(-6) M. The pH optimum for the reaction was approximately 7.5. Blood samples from 73 randomly selected adult subjects had a mean activity of 10.2 +/- 2.4 (mean +/- S.D.) units/ml packed red blood cells. The range of activities was from 4.6 to 14.2 units/ml. The results of experiments in which partially purified human kidney TPMT was added to RBC lysates and of experiments in which "low" and "high" activity lysates were mixed gave no indication that individual variations in RBC TPMT activity were due to endogenous inhibitors or activators of the enzyme.


Current Indications for Transoral Robotic Surgery in Oropharyngeal Cancer.
The incidence of oropharyngeal squamous cell carcinoma (OPSCC) is increasing dramatically and is conclusively linked to increasing rates of human papillomavirus (HPV) infection. HPV-related oropharyngeal cancers have been shown to occur in a unique demographic group and show favorable oncologic outcomes compared with HPV-negative OPSCC. There has been a paradigm shift in the treatment of early-stage OPSCC, with most patients now undergoing primary surgery in the United States. Transoral robotic surgery is associated with excellent oncologic and functional outcomes in the treatment of OPSCC and is increasingly being used for a broader range of oropharyngeal indications.


Systematic review and meta-analysis of randomized clinical trials of self-expanding metallic stents as a bridge to surgery versus emergency surgery for malignant left-sided large bowel obstruction.
Use of self-expanding metallic stents (SEMS) as a bridge to surgery has been suggested as an alternative management for acute malignant left-sided colonic obstruction, as emergency surgery has a high risk of morbidity and mortality. This meta-analysis evaluated high-quality evidence comparing preoperative SEMS with emergency surgery. Relevant randomized clinical trials (RCTs) were identified from the Cochrane Central Register of Controlled Trials, MEDLINE, Embase and PubMed (1990-2011). Primary outcomes were primary anastomosis, stoma and in-hospital mortality rates. Secondary outcomes included anastomotic leak, 30-day reoperation and surgical-site infection rates. Four RCTs with 234 patients were included. Technical and clinical success rates for stenting were 70·7 per cent (82 of 116) and 69·0 per cent (80 of 116) respectively. The clinical perforation rate was 6·9 per cent (8 of 116) and the silent perforation rate 14 per cent (11 of 77). SEMS intervention resulted in significantly higher successful primary anastomosis (risk ratio (RR) 1·58, 95 per cent confidence interval 1·22 to 2·04; P < 0·001) and lower overall stoma (RR 0·71, 0·56 to 0·89; P = 0·004) rates. There was no difference in primary anastomosis, permanent stoma, in-hospital mortality, anastomotic leak, 30-day reoperation and surgical-site infection rates. Three trials were stopped prematurely, one because the emergency surgery group had a significantly increased anastomotic leak rate, and two others because of stent-related complications and increased 30-day morbidity following SEMS management. Technical and clinical success rates for stenting were lower than expected. SEMS is associated with a high incidence of clinical and silent perforation. However, as a bridge to surgery, SEMS has higher successful primary anastomosis and lower overall stoma rates, with no significant difference in complications or mortality.


Cytochrome b positive X-linked chronic granulomatous disease: a normal cell surface expression of cytochrome b.
The polymorphonuclear (PMN) cells from a patient with cytochrome b positive X-linked chronic granulomatous disease (Xb+ CGD) were studied using flow cytometry. Both the cell surface expression of monoclonal antibody defined cytochrome b and the superoxide production (intracellular 2',7'-Dichlorofluorescin Diacetate oxidation) were investigated at a single cell level. Flow cytometry clearly demonstrated the complete absence of superoxide production in the patient's PMN cells, the mosaicism in his mother's PMN cells and also indicated the normal cell surface expression of cytochrome b. The results obtained by Western blot analysis and reduced-minus-oxidized spectra confirmed the presence of functional and normal amounts of cytochrome b. We concluded that this is a case of Xb+ CGD with a normal cell surface expression of cytochrome b.


Saccharomyces cerevisiae KTR4, KTR5 and KTR7 encode mannosyltransferases differentially involved in the N- and O-linked glycosylation pathways.
Saccharomyces cerevisiae is a model to understand basic aspects of protein glycosylation pathways. Although these metabolic routes have been thoroughly studied, there are still knowledge gaps; among them, the role of the MNT1/KRE2 gene family. This family is composed of nine members, with only six functionally characterized. The enzymes Ktr1, Ktr3, and Mnt1/Kre2 have overlapping activities in both O-linked and N-linked glycan synthesis; while Ktr2 and Yur1 participate exclusively in the elongation of the N-linked glycan outer chain. KTR6 encodes for a phosphomannosyltransferase that synthesizes the cell wall phosphomannan. Here, we aimed to establish the functional role of KTR4, KTR5 and KTR7 in the protein glycosylation pathways, by using heterologous complementation in Candida albicans null mutants lacking members of the MNT1/KRE2 gene family. The three S. cerevisiae genes restored defects in the C. albicans N-linked glycosylation pathway. KTR5 and KTR7 partially complemented a C. albicans null mutant with defects in the synthesis of O-linked glycans, and only KTR4 fully elongated the O-linked glycans like wild-type cells. Therefore, our results suggest that the three genes have a redundant activity in the S. cerevisiae N-linked glycosylation pathway, but KTR4 plays a major role in O-linked glycan synthesis.


Taxifolin-3-O-glucoside from Osbeckia nepalensis Hook. mediates antihyperglycemic activity in CC1 hepatocytes and in diabetic Wistar rats via regulating AMPK/G6Pase/PEPCK signaling axis.
Osbeckia nepalensis Hook. f. is an ICMR documented plant well known for its antidiabetic uses among the folk people of Northeast Region of India. In-depth study with scientific substantiation of the plant may uphold the therapeutic potential against the treatment of type 2 diabetes mellitus (T2DM). The present study evaluates the traditionally claimed prophylactic potential of O. nepalensis and its extracts along with the isolated compound taxifolin-3-O-glucoside (TG) against the downregulation of T2DM related hepatic gluconeogenesis through in vitro, in vivo and in silico conditions as a means of ameliorating hyperglycemia. Antidiabetic potential of O. nepalensis was carried out in both CC1 hepatocytes (in vitro) and STZ-induced diabetic male Wistar rats (in vivo). Enriched bioactive fraction and bioactive molecules were isolated through bioactivity-guided fractionation, yielding two major molecules, taxifolin-3-O-glucoside and quercitin-3-O-rhamnoside. The bioactivity of taxifolin-3-O-glucoside was validated through immunoblotting techniques aided by in silico molecular docking and simulations. Methanolic extract of O. nepalensis and taxifolin-3-O-glucoside (TG) isolated thereof enhanced the uptake of glucose in CC1 hepatocytes and downregulates the gluconeogenic enzymes (G6Pase and PEPCK) and its related transcription factors (FOXO1, HNF4α and PGC1α) through the stimulation of AMPK phosphorylation in in vitro condition. Moreover, in in vivo experiments, the in vitro most active fraction BuSFr1 (consisting of the two active major compounds taxifolin-3-O-glucoside and quercitin-3-O-rhamnoside) exhibited a substantial decrease in elevated blood glucose level and increase the glucose tolerance as well as plasma insulin level. In silico molecular docking and simulations for TG with the protein G6Pase inferred the docking sites and stability and showed taxifolin-3-O-glucoside as more potent and non-toxic as compared to quercitin-3-O-rhamnoside. The traditionally claimed antidiabetic effect of O. nepalensis has been proved to be effective in lowering the blood glucose level through in vitro, in vivo and in silico analysis which will pave a way for the development of antidiabetic phytopharmaceutical drugs which can be validated through further clinical studies.


Integral approach for the evaluation of poultry manure, compost, and digestate: Amendment characterization, mineralization, and effects on soil and intensive crops.
The egg industry has increased its production worldwide during the last decades. Several waste management strategies have been proposed to treat large volumes of poultry manure. Composting and anaerobic digestion are the main stabilization processes used. However, there are disagreements on the criteria for applying raw and treated poultry manure to the soil. We studied the relationship between physicochemical, toxicological, microbiological, parasitological, and metabarcoding parameters of raw and treated poultry manure (compost and digestate). Subsequently, we evaluated the mineralization of C, N and P, and the effects of amended soil on horticultural and ornamental crops. Compost and digestate presented better general conditions than poultry manure for use as organic soil amendments. The highest pathogenic microorganism content (total and fecal coliforms, Escherichia coli, and Salmonella spp.) was recorded for poultry manure. Multivariate analyses allowed associating a lower phytotoxicity with compost and a higher microbial diversity with digestate. Therefore, only compost presented stability and maturity conditions. We found high released CO2-C, N loss, and P accumulation in soil amended with a high dose of poultry manure during mineralization. However, high doses of poultry manure and digestate increased the biomass production in the valorization assay. We recommend the soil application of stabilized and mature poultry manure-derived amendments, which reduce the negative impacts on the environment and promote more sustainable practices in agricultural systems.


Pediatric issues with cavovarus foot deformities.
Cavovarus foot deformity in children has numerous etiologies with a general pathophysiologic mechanism of muscle imbalance. It is of great importance in the evaluation of a child with a cavovarus foot to determine the underlying cause of the deformity, as the most common origin is a progressive neurologic condition that may be complicated by other orthopedic problems. Treatment options typically are surgical, with limited indications for nonsurgical modalities, and must consider the age of the patient, the nature of the neurologic disease, and the severity of the deformity. Current surgical procedures can be divided into soft tissue procedures to rebalance the muscle forces, osteotomies, and triple arthrodesis. Triple arthrodesis is considered a salvage procedure reserved for the older child with severe, rigid deformity who has failed other surgical treatments.


Tissue specificity in the expression of Friend erythroleukemic virus sequences in infected mouse tissues.
The SQA cell line produces Friend leukemia virus that remains leukemogenic after serial passages in vitro. The state of the provirus and its expression were investigated in newborn and adult mouse tissues, using probes specific for ecotropic and xenotropic sequences. Genomic ecotropic and xenotropic sequences were similar in size in spleen and liver of infected and control animals but appear amplified in infected tissues. Expression of these sequences however differed. Several species of xenotropic and ecotropic-specific RNAs were detected in infected spleens, in SQA cells and in the liver of newborn infected animals but were absent in infected adult liver and control tissues. These results suggest that activation and expression of ecotropic and xenotropic endogenous sequences may play a role in pathogenesis of the disease.


Factors influencing uptake of genetic testing for colorectal cancer risk in an Australian Jewish population.
There is a significant excess of colorectal cancer in the Australian Ashkenazi Jewish community. This excess can partially be attributed to inherited factors that are over represented in this population, such as the APC variant I1307K, which is associated with a modest increase in colorectal cancer risk. There is currently only sporadic clinical genetic testing offered for this variant, as neither the exact increase in cancer risk and therefore the appropriate screening strategies for I1307K carriers, nor the acceptability of such testing in Jewish communities have been determined. This study reports a high acceptability of such genetic testing within a community sample of 300 Australian Jewish individuals--94% of participants would have a test for predisposition to colorectal cancer and a majority would make this decision based on the desire for information for their families and to decrease their own cancer risk. Some concerns were noted about genetic testing for cancer predisposition, including insurance discrimination, test accuracy and confidentiality.


Effects of Breathe Right on snoring: a polysomnographic study.
The nasal vestibule is a major site of resistance to airflow in healthy subjects. A high nasal resistance may increase snoring. Activation of the alae nasi and alar retraction reduce resistance to airflow and improve ventilation. The Breathe Right (BR) device has been proposed to reduce or eliminate snoring by improving nasal breathing. We assessed the efficacy of BR on sleep quality and snoring during 2 full-night polysomnographies, the first without and the second with BR. Ten non-apnoeic snorers were studied. Snoring was present during 22-98% of total sleep time during the control night. Ear-nose-throat examination disclosed a nasal valve anomaly in five subjects, objectivated by anterior and posterior rhinomanometry. Quality of sleep and snoring were not influenced by BR, even when different sleep stages were analysed separately. No difference in snoring index was found between snorers with or without nasal valve anomaly. We conclude that BR is ineffective in relieving snoring in non-apnoeic snorers.


The potential influence of interferon-α on the growth of the growing teratoma: A case report.
Growing teratoma syndrome (GTS) is rare. It is fatal if the teratoma is unresectable. A standard systemic therapy is not established. The efficacy of interferon-alpha (IFN-α) for GTS was described but the treatment periods were relatively short. A 23-year-old Japanese male with bulky retroperitoneal lymph node and multiple lung metastases that progressed to GTS was administered 6 × 106 units of natural IFN-α 2 × /week. Since the IFN-α treatment suppressed both lesions' growth, it was continued for >10 years. The patient is well with controlled metastases (135 months since the IFN-α's initiation). This is apparently the longest follow-up of INF-α treatment for GTS.


Characterization of plant Aurora kinases during mitosis.
The Aurora kinase family is a well-characterized serine/threonine protein kinase family that regulates different processes of mitotic events. Although functions of animal and yeast Aurora kinases have been analyzed, plant aurora kinases were not identified and characterized. We identified three Aurora kinase orthologs in Arabidopsis thaliana and designated these as AtAUR1, AtAUR2, and AtAUR3. These AtAURs could phosphorylate serine 10 in histone H3, in vitro. Dynamic analyses of GFP-fused AtAUR proteins revealed that AtAUR1 and AtAUR2 localized at the nuclear membrane in interphase and located in mitotic spindles during cell division. AtAUR1 also localized in the cell plates. AtAUR3 showed dot-like distribution on condensed chromosomes at prophase and then localized at the metaphase plate. At late anaphase, AtAUR3 is evenly localized on chromosomes. The localization of AtAUR3 during mitosis is very similar to that of phosphorylated histone H3. Interestingly, an overexpression of AtAUR3 induces disassembly of spindle microtubules and alteration of orientation of cell division. Our results indicate that plant Aurora kinases have different characters from that of Aurora kinases of other eukaryotes.


Connecting the dots in ethology: applying network theory to understand neural and animal collectives.
A major goal shared by neuroscience and collective behavior is to understand how dynamic interactions between individual elements give rise to behaviors in populations of neurons and animals, respectively. This goal has recently become within reach, thanks to techniques providing access to the connectivity and activity of neuronal ensembles as well as to behaviors among animal collectives. The next challenge using these datasets is to unravel network mechanisms generating population behaviors. This is aided by network theory, a field that studies structure-function relationships in interconnected systems. Here we review studies that have taken a network view on modern datasets to provide unique insights into individual and collective animal behaviors. Specifically, we focus on how analyzing signal propagation, controllability, symmetry, and geometry of networks can tame the complexity of collective system dynamics. These studies illustrate the potential of network theory to accelerate our understanding of behavior across ethological scales.


Cadence-based classification of moderate-intensity overground walking in 41- to 85-year-old adults.
Walking cadence (steps/min) has emerged as a valid proxy of physical activity intensity, with consensus across numerous laboratory-based treadmill studies that ≥100 steps/min approximates absolutely defined moderate intensity (≥3 metabolic equivalents; METs). We recently reported that this cadence threshold had a classification accuracy of 73.3% for identifying moderate intensity during preferred pace overground walking in young adults. The purpose of this study was to evaluate and compare the performance of a cadence threshold of ≥100 steps/min for correctly classifying moderate intensity during overground walking in middle- and older-aged adults. Participants (N = 174, 48.3% female, 41-85 years of age) completed laboratory-based cross-sectional study involving an indoor 5-min overground walking trial at their preferred pace. Steps were manually counted and converted to cadence (total steps/5 min). Intensity was measured using indirect calorimetry and expressed as METs. Classification accuracy (sensitivity, specificity, accuracy) of a cadence threshold of ≥100 steps/min to identify individuals walking at ≥3 METs was calculated. The ≥100 steps/min threshold demonstrated accuracy of 74.7% for classifying moderate intensity. When comparing middle- vs. older-aged adults, similar accuracy (73.4% vs. 75.8%, respectively) and specificity (33.3% vs. 34.5%) were observed. Sensitivity was high, but was lower for middle- vs. older-aged adults (85.2% vs. 93.9%, respectively). A cadence threshold of ≥100 steps/min accurately identified moderate-intensity overground walking. Furthermore, accuracy was similar when comparing middle- and older-aged adults. These findings extend our previous analysis in younger adults and confirm the appropriateness of applying this cadence threshold across the adult lifespan.


Parametric analysis of surfactant-aided imbibition in fractured carbonates.
Many carbonate oil reservoirs are oil-wet and fractured; waterflood recovery is very low. Dilute surfactant solution injection into the fractures can improve oil production from the matrix by altering the wettability of the rock to a water-wetting state. A 2D, two-phase, multicomponent, finite-volume, fully-implicit numerical simulator calibrated with our laboratory results is used to assess the sensitivity of the process to wettability alteration, IFT reduction, oil viscosity, surfactant diffusivity, matrix block dimensions, and permeability heterogeneity. Capillarity drives the oil production at the early stage, but gravity is the major driving force afterwards. Surfactants which alter the wettability to a water-wet regime give higher recovery rates for higher IFT systems. Surfactants which cannot alter wettability give higher recovery for lower IFT systems. As the wettability alteration increases the rate of oil recovery increases. Recovery rate decreases with permeability significantly for a low tension system, but only mildly for high tension systems. Increasing the block dimensions and increasing oil viscosity decreases the rate of oil recovery and is in accordance with the scaling group for a gravity driven process. Heterogeneous layers in a porous medium can increase or decrease the rate of oil recovery depending on the permeability and the aspect ratio of the matrix block.


Melanoma and rheumatoid arthritis (brief report).
The aim of this study is to assess melanoma risk in rheumatoid arthritis (RA). A literature review was performed, retrieving observational studies over 1990-2010 that provided estimates of relative risk of melanoma associated with RA, compared to the general population. We generated standardized incidence ratio (SIR) estimates across all studies, first pooling the data and then performing a random-effects model to generate the SIRs. We retrieved 713 citations; after reviewing the titles and abstracts, 124 were further reviewed, and of these, 11 met our inclusion criteria for analysis. Pooling the data, there were a total of 601 melanomas that were observed over 1,351,061 patient-years of follow-up, or 4.4 cases per 10,000 years. The expected number of melanomas over this interval was 596.2 for a pooled SIR of 1.01 [95% confidence interval (CI), 0.93, 1.09]. Excluding the two patient groups that were known to be exposed to biologics, the SIR estimate was unchanged (1.01; 95 CI, 0.93, 1.10). Our random-effects model similarly indicated an SIR for melanoma of 0.95 [95% credible interval (CrI), 0.86, 1.03] overall and 0.95 (95% CrI, 0.86, 1.04) excluding the two patient groups that were known to be exposed to biologics. These results do not highlight an important increased risk for melanoma in RA patients over-all, compared to the general population. It is not clear whether the risk is different for patients specifically exposed to biologic agents, although data are relatively few. Further study, especially of RA patients with a past history of melanoma, is warranted.


The Diversity of Small Mammals along a Large River Valley Revealed from Pellets of Tawny Owl Strix aluco.
The Vistula River is one of the largest European semi-natural rivers of high ecological value that functions as an ecological corridor. To assess the structure of small mammal communities along the Vistula River, an analysis of the diet of an opportunistic predator, the tawny owl Strix aluco, was used. A total of 6355 individuals of 19 species were found, including 5 soricomorph species, 12 rodents, 1 carnivore, and 1 bat species. Tawny owls most frequently caught Apodemus agrarius, Clethrionomys glareolus, Apodemus flavicollis, and Microtus arvalis. Rodents dominated small mammal communities (90%), followed by soricomorphs (8%), and the share of Chiroptera was significant (2%). Using Ward's method in cluster analysis, three clusters of sites with similar mammal communities were identified. The cluster that included 17 study sites with the dominance of agriculture habitats was inhabited by diverse mammal communities with a high number of species. In the cluster composed of three suburban forest sites, mammal communities had the lowest diversity, although the high species richness and the highest shares of the forest species (A. flavicollis, C. glareolus, and Nyctalus noctula). Mammal communities in the cluster were composed of three urban sites were dominated by A. agrarius and M. arvalis. The study indicates the high species richness of small mammals in floodplains of the Vistula River and the adjacent areas in central Poland. The floodplain offers suitable habitats for species associated with forests, water bodies, agricultural land, and developed areas. The data collected confirms earlier model predictions about the presence of well-connected local populations of forest mammals along the Vistula River.


Seed size selection in heteromyids : A second look.
The general conclusion of this paper is that heteromyids do not select seed sizes on the basis of their body size. My conclusion comes from the analysis of new data from central New Mexico, analysis of data in the literature on food habits of heteromyids, and a reanalysis of Brown and Lieberman (1973) and Brown (1975). All of these sources agree that no seed size selection exists.Although no pattern of simple seed size selection was found, interesting differences were noticed among species. First, the tendency to husk appears to be related to the size of the rodent. Second, large heteromyids may depend on fruiting heads made up of small seeds. This may or may not have significance to the coexistece of heteromyid communities. At present, insufficient data are available to make conclusions along these lines.Intuitively satisfying hypotheses, such as seed size allocation by heteromyids, are normally very hard to lay to rest. It is my hope that this paper demonstrates the weight of evidece is against seed size allocation in heteromyids. Those who wish to maintain this hypothesis, or reveal that it or related hypotheses have anything to do with heteromyid coexistence, must now produce data to support their position.


Functional neuroanatomy of the basal ganglia as studied by dual-probe microdialysis.
Dual probe microdialysis was employed in intact rat brain to investigate the effect of intrastriatal perfusion with selective dopamine D1 and D2 receptor agonists and with c-fos antisense oligonucleotide on (a) local GABA release in the striatum; (b) the internal segment of the globus pallidus and the substantia nigra pars reticulata, which is the output site of the strionigral GABA pathway; and (c) the external segment of the globus pallidus, which is the output site of the striopallidal GABA pathway. The data provide functional in vivo evidence for a selective dopamine D1 receptor-mediated activation of the direct strionigral GABA pathway and a selective dopamine D2 receptor inhibition of the indirect striopallidal GABA pathway and provides a neuronal substrate for parallel processing in the basal ganglia regulation of motor function. Taken together, these findings offer new therapeutic strategies for the treatment of dopamine-linked disorders such as Parkinson's disease, Huntington's disease, and schizophrenia.


Adenosine deaminase production by an endophytic bacterium (Lysinibacillus sp.) from Avicennia marina.
The present study was carried out with the following objectives: (1) to isolate the endophytic bacilli strains from the leaves of mangrove plant Avicennia marina, (2) to screen the potential strains for the production of adenosine deaminase, (3) to statistically optimize the factors that influence the enzyme activity in the potent strain, and (4) to identify the potent strain using 16S rRNA sequence and construct its phylogenetic tree. The bacterial strains isolated from the fresh leaves of a mangrove A. marina were assessed for adenosine deaminase activity by plating method. Optimization of reaction process was carried out using response surface methodology of central composite design. The potent strain was identified based on 16S rRNA sequencing and phylogeny. Of five endophytic strains, EMLK1 showed a significant deaminase activity over other four strains. The conditions for maximum activity of the isolated adenosine deaminase are described. The potent strain EMLK1 was identified as Lysinibacillus sp. (JQ710723) being the first report as a mangrove endophyte. Mangrove-derived endophytic bacillus strain Lysinibacillus sp. EMLK1 is proved to be a promising source for the production of adenosine deaminase and this enzyme deserves further studies for purification and its application in disease diagnosis.


Studies on pituitary follitropin. X. Biochemical, receptor binding and immunological properties of deglycosylated ovine hormone.
Treatment of ovine pituitary follitropin with anhydrous hydrogen fluoride at 0 degrees C for 60 min resulted in the removal of approximately 80% of the sugars without affecting the polypeptide moiety. Fucose, sialic acid and N-acetylgalactosamine were eliminated completely while the loss of hexoses and N-acetylglucosamine amounted to 86% and 56% respectively. The treatment had no effect on the quaternary structure of the hormone. Consistent with the loss of carbohydrate, the elution volume of the hormone on Sephadex G100 was increased and the deglycosylated hormone lost its ability to interact with the lectin concanavalin-A immobilized on Sepharose. Deglycosylation rendered the hormone less acidic as reflected by altered electrophoretic mobility in polyacrylamide gels at pH 8.9 and 4.5. The UV absorption spectrum of the hormone was not affected by deglycosylation but tryptophan fluorescence was slightly decreased. The receptor-binding activity of the hormone as estimated by a specific radioreceptor assay using adult bovine testicular membranes and labeled 125I-FSH was increased by 250% following deglycosylation. In a conformation specific radioimmunoassay, the deglycosylated hormone showed 150% activity as compared to the native hormone. Thus, the full integrity of the carbohydrate moiety in ovine follitropin is not required for effective receptor binding and immunological activity.


Data in intelligent approach for estimation of disc cutter life using hybrid metaheuristic algorithm.
This data in brief presents the monitoring data measured during shield tunnelling of Guangzhou-Shenzhen intercity railway project. The monitoring data includes shield operational parameters, geological conditions, and geometry at the site. The presented data were arbitrarily split into two subsets including the training and testing datasets. The field observations are compared to the forecasting values of the disc cutter life assessed using a hybrid metaheuristic algorithm proposed for "Prediction of disc cutter life during shield tunnelling with artificial intelligent via incorporation of genetic algorithm into GMDH-type neural network" [1]. The presented data can provide a guidance for cutter exchange in shield tunnelling.


Perceptions of health professionals on subcutaneous hydration in palliative care: A qualitative study.
Evidence indicates that hypodermoclysis is as safe and effective as intravenous rehydration in the treatment of the symptomatology produced by mild to moderate dehydration in patients for whom oral route administration is not possible. However, the knowledge about the use of the subcutaneous hydration and its correlates is still limited. To explore the perceptions, attitudes and opinions of health professionals in palliative care on the administration of subcutaneous hydration. This is a qualitative focus group study with health professionals of palliative care. Four focus groups were carried out until data saturation. A qualitative content analysis was performed. A total of 37 participants, physicians and nurses, were recruited from different services of palliative care in Spain. In all, 856 meaning units were identified, from which 56 categories were extracted and grouped into 22 sub-themes, which were distributed among four themes: 'factors which influence the hydration decision', 'factors related to the choice of the subcutaneous route for hydration', 'the subcutaneous hydration procedure' and 'performance guidelines and/or protocols'. Variables which most often influence the use of subcutaneous route to hydration are those that are linked to the characteristics of the patient, the team and the family, and other like the context and professionals' subjective perceptions about this medical practice.