Combining a Simple Method for DNA/RNA/Protein Co-Purification and Arabidopsis Protoplast Assay to Facilitate Viroid Research.
Plant-viroid interactions represent a valuable model for delineating structure-function relationships of noncoding RNAs. For various functional studies, it is desirable to minimize sample variations by using DNA, RNA, and proteins co-purified from the same samples. Currently, most of the co-purification protocols rely on TRI Reagent (Trizol as a common representative) and require protein precipitation and dissolving steps, which render difficulties in experimental handling and high-throughput analyses. Here, we established a simple and robust method to minimize the precipitation steps and yield ready-to-use RNA and protein in solutions. This method can be applied to samples in small quantities, such as protoplasts. Given the ease and the robustness of this new method, it will have broad applications in virology and other disciplines in molecular biology.


The evolution of volatile compounds profile of "Toscano" dry-cured ham during ripening as revealed by SPME-GC-MS approach.
The volatile compounds profile is an important feature for the characterization of dry-cured hams. Some minor typical Italian products, such as 'Toscano' ham, have been poorly studied in regards to their composition of volatile compounds. In this article, we studied the evolution of the aromatic profile of 'Toscano' dry-cured ham by solid-phase microextraction-gas chromatographic-mass spectrometry (SPME-GC-MS) with ripening. Ten right thighs were cured according to the 'Toscano' PDO protocol, sampled at 0, 1, 3, 6 and 12 months and submitted to volatile compounds analysis by SPME with a Divinylbenzene (DVB)/Carboxen/Polydimethylsiloxane (PDMS) 75-µ Stable Flex fibre. An Agilent 5975C mass selective detector (MSD) spectrometer with electron ionization (EI) source operating in scan mode within the m/z 29-350 range was used for data collection. Seven internal standards, either deuterium labeled or absent in the specimens and chosen to represent low or high boiling esters, alcohols, acids or phenols, were added to the homogenized samples and used to normalize the SPME fibre response to account for response changes upon wearing. Linear calibrations were obtained in this way for selected representative compounds. Over 60 compounds belonging to esters, aldehydes, organic acids, ketones and alcohols were identified by comparison with spectral libraries and Kovats indices. Aldehydes were the most represented chemical family, followed by organic acids, alcohols, ketones and esters. The aldehydes and ketones increased during the first 3 months, when the larger formation of volatiles occurred. For other families, the evolution over time was less evident. The principal component and discriminant analyses of the aromatic profile were effective in classifying the hams at 0, 6 or 12 months of ripening while for 1 and 3 months' samples a partial overlapping was shown. These results represent the first characterization of 'Toscano' ham and may constitute the basis to identify the best ripening time and define an analytical quality standard for this typical ham.


Clinical and bacteriologic evaluation of BACTEC resin-containing blood culture medium.
In a retrospective study of 1,143 blood culture sets, BACTEC aerobic (6B) and osmotically stabilized (8B) media were compared individually with resin-containing (16B) medium for the isolation of bacteria from the blood of patients receiving antimicrobial therapy. The 16B medium was found to detect significantly more positive cultures than either 6B (P less than 0.01) or 8B (P less than 0.001). For 22 of 25 isolates of Staphylococcus aureus from 18 patients, 16B medium provided the only means of recovery. All but one of these patients were receiving appropriate antimicrobial therapy, and 16 of 18 had a previous blood culture set positive for S. aureus that did not include a 16B bottle. There was no evidence of a change in antimicrobial therapy in response to a 16B positive culture in these patients. No significant increase in the recovery of other gram-positive or gram-negative bacteria or decrease in the time to radiometric detection of positive cultures as a result of using 16B medium was noted.


The application of novel hydrophobic ionic liquids to the extraction of uranium(VI) from nitric acid medium and a determination of the uranyl complexes formed.
Novel ammonium based hydrophobic ionic liquids (ILs) have been synthesised and characterised, and their use in the liquid-liquid extraction of uranium(VI) from an aqueous nitric acid solution using tri-n-butyl phosphate (TBP), studied. On varying the nitric acid concentration, each IL was found to give markedly different results. Relatively hydrophilic ILs showed high uranium(VI) extractability at 0.01 M nitric acid solution which progressively decreased from 0.01 to 2 M HNO(3) and then increased again as the nitric acid concentration was increased to 6 M. An analysis of the mechanisms involved for one such IL, pointed to cationic-exchange being the predominant route at low nitric acid concentrations whilst at high nitric acid concentrations, anionic-exchange predominated. Strongly hydrophobic ILs showed low extractability for nitric acid concentrations below 0.1 M but increasing extractability from 0.1 M to 6 M nitric acid. The predominant mechanism in this case involved the partitioning of a neutral uranyl complex. The uranyl complexes were found to be UO(2)(2+)·(TBP)(3) for the cationic exchange mechanism, UO(2)(NO(3))(2)(TBP)(2) for the neutral mechanism and UO(2)(NO(3))(3)(-)·(TBP) for the anionic exchange mechanism.


[A clinicopathological study of IgG4-related ocular disease and differentiated diagnosis with non-IgG4-related ocular disorders].
To study the clinicopathologic features of IgG4-related ocular disease and differentiated diagnosis with non-IgG4-related ocular disorders. Forty-three cases were selected from the Department of Pathology, West China Hospital, Sichuan University from December 2008 to January 2015 with retrospective series case study. The clinical manifestation, laboratory examination, imaging findings, treatment and histopathological characteristics were analyzed. According to the diagnostic criteria for IgG4-related ocular disease in 2014 and relevant literatures, our cases were divided into two groups: IgG4-related ocular disease (23 samples from 18patients) and non-IgG4-related ocular disease (31 samples from 25 patients). The male-to-female ratio, the periductal fibrosis and the orbital lesion area was analyzed by Pearson chi-square. The Immunohistochemistry of IgG4+/HPF, IgG+/HPF and IgG4+/IgG+ ratio and age was analyzed by two sample t test. The involved structural tissue and the infiltration of eosinophils and lymphocytes and plasma cells was analyzed by signed-rank test. The significant statistic differences were observed between these two groups in immunohistochemical staining of IgG4 (>40/HPF) (t=12.17, P<0.01), IgG (t=2.91, P<0.01) and IgG4+/IgG+ ratio (≥40%) (t=16.98, P<0.01), the lymphocytic and plasmacytic infiltration (F=3.62, P=0.03),the periductal fibrosis(F=4.71 , P=0.03), the involved region of orbit (t=1.11, P=0.01), but not in the mean age and male-to-female ratio (t=1.09, P=0.58). IgG4 ROD should be distinguished form the non-specific inflammation disorders, lymphoma and so on. IgG4-related ocular disease often arises from middle-old man, characterized by bilateral orbital painless swelling and abundant IgG4+ plasma cells infiltration. Nearly all orbital tissue could be involved by IgG4-related ocular disease, especially lacrimal gland. But the characteristic obliterating phlebitis were not seen. The immunohistochemical staining of IgG4 and IgG plays an important role in differentiating IgG4-related ocular disease from non-IgG4-related ocular disorders. Combining clinical manifestation, laboratory examination, imaging findings with histopathology is helpful for diagnosis and differential diagnosis. (Chin J Ophthalmol, 2016, 52: 256-262).


Cross-talk between microbiota and immune fitness to steer and control response to anti PD-1/PDL-1 treatment.
Immune Checkpoint Inhibitors (ICIs) are improving the survival of cancer patients, however only the 20-30% of treated patients present clinical benefits. Toxicity represents the major cause of reduced dosage, delayed drug administration and therapy discontinuation. Hence in the context of multiple treatment possibilities, the identification of predictive markers of response and toxicity is a challenging approach for drug selection in order to obtain the best clinical benefit while minimizing the side effects. The loss of the protective function of intestinal barriers that interacts with the environment measured as increased intestinal permeability and the changes occurring in the microbiota composition have been proposed as a mechanism potentially explaining the pathogenesis of immune related toxicity.In this review we discuss the new perspectives on the involvement of PD-1 and PDL-1 in the cross talk between gut microbiota and immune fitness and how gut microbiota impacts on the efficacy of anti-PD-1 and anti-PDL-1 treatments in cancer.


Development and validation of real-time quantitative reverse transcriptase-polymerase chain reaction for monitoring gene expression in cardiac myocytes in vitro.
In this article we present validation of a real-time RT-PCR method to quantitate mRNA expression levels of atrial natriuretic peptide and c-fos in an in vitro model of cardiac hypertrophy. This method requires minimal sample and no postreaction manipulation. In real-time RT-PCR a dual-labeled fluorescent probe is degraded concomitant with PCR amplification. Input target mRNA levels are correlated with the time (measured in PCR cycles) at which the reporter fluorescent emission increases beyond a threshold level. The use of an oligo(dt) magnetic bead protocol to harvest poly(A) mRNA from cultured cells in 96-well plates minimized DNA contamination. We show that the GAPDH gene chosen for normalization of the RNA load is truly invariant throughout the biological treatments examined. We discuss two methods of calculating fold increase: a standard curve method and the DeltaDelta Ct method. Real-time quantitative RT-PCR was used to determine the time course of c-fos induction and the effect of varying doses of four known hypertrophy agents on atrial naturitic factor messenger RNA expression in cultured cardiac muscle cells. Our results agree with published data obtained from Northern blot analysis.


Individual effects of sodium, potassium, calcium, and magnesium chloride salts on Lactobacillus pentosus and Saccharomyces cerevisiae growth.
A quantitative investigation on the individual effects of sodium (NaCl), potassium (KCl), calcium (CaCl2), and magnesium (MgCl2) chloride salts against Lactobacillus pentosus and Saccharomyces cerevisiae, two representative microorganisms of table olives and other fermented vegetables, was carried out. In order to assess their potential activities, both the kinetic growth parameters and dose-response profiles in synthetic media (deMan Rogosa Sharpe broth medium and yeast-malt-peptone-glucose broth medium, respectively) were obtained and analyzed. Microbial growth was monitored via optical density measurements as a function of contact time in the presence of progressive chloride salt concentrations. Relative maximum specific growth rate and lag-phase period were modeled as a function of the chloride salt concentrations. Moreover, for each salt and microorganism tested, the noninhibitory concentrations and the MICs were estimated and compared. All chloride salts exerted a significant antimicrobial effect on the growth cycle; particularly, CaCl2 showed a similar effect to NaCl, while KCl and MgCl2 were progressively less inhibitory. Microbial susceptibility and resistance were found to be nonlinearly dose related.


Clinical profile and outcome of swine flu in Indian children.
To describe the clinical characteristics and outcome of Indian children infected with 2009 H1N1 influenza virus. Retrospective chart review. Outpatient department and hospitalized patients in a tertiary care hospital. Clinical details of 85 children (positive for the 2009 H1N1 virus infection tested by real time reverse transcriptas epolymerasechain reaction assay) were analyzed from medical charts. Of the 85 (55 boys) children positive for 2009 H1N1 virus infection, 64.7%were between 5 years to 16 years, and 35.3%were below 5 years age. The mean age of these children was 7.5±3.5 yr. Contact history was positive only in 22 (26%) cases. High grade fever was the most common symptom, followed by cough and rhinorrhea. Twenty-nine (34%) patients had an underlying co-morbid condition. Of the 34 patients who underwent chest radiography during evaluation, 18 children (52.9%) had findings consistent with lower respiratory tract infection. Antiviral therapy was initiated in 76 patients. Hospitalization was required in 30 (35.3%) children. Risk factors for hospitalization included underlying co-morbid condition, respiratory distress, vomiting, wheezing, diarrhea, hypotension and infiltrates/consolidation on chest radiograph. Mean length of hospitalization was 131±76 hours, irrespective of underlying disease. Three children developed Acute Respiratory Distress Syndrome and died. Clinical features and routine laboratory investigations in children with swine origin influenza were non-specific. Children with co-morbid condition, respiratory distress, vomiting, wheezing, diarrhea, hypotension and infiltrates/consolidation on chest radiograph were at higher risk of hospitalization.


The influence of CT and dual-energy X-ray absorptiometry (DXA) bone density on quantitative [18F] sodium fluoride PET.
[18F] sodium fluoride PET/CT provides quantitative measures of bone metabolic activity expressed by the parameters standardised uptake value (SUV) and bone plasma clearance (K i) that correlate with measurements of bone formation rate obtained by bone biopsy with double tetracycline labelling. Both SUV and K i relate to the tracer uptake in each millilitre of tissue. In general, the bone region of interest (ROI) includes both mineralised bone {generally with a high concentration of [18F]NaF} and bone marrow (with a much lower concentration), suggesting that correcting SUV and K i for volumetric bone mineral density (vBMD) and measuring them with respect to the tracer uptake in each gram of bone mineral might improve the correlation with the findings of bone biopsy. As a first test of this hypothesis, we looked for positive correlations between SUV and K i values with CT and DXA bone mineral density (BMD) parameters measured in the same ROI. A retrospective reanalysis was performed of 63 lumbar spine [18F]NaF PET/CT scans acquired in four earlier studies. The quantitative PET parameters SUV and K i were measured in L1-L4 and Hounsfield units (HU) measured on the CT scans in the same ROI. Spine BMD data was also obtained from DXA scans in the form of areal BMD and used to derive the bone mineral apparent density (BMAD, an estimate of vBMD). Scatter plots were drawn of SUV and K i against HU, BMAD and areal BMD and the Spearman rank correlation coefficients derived for each plot. All correlations were positive and statistically significant. Correlations were highest for HU (SUV: RS =0.513, P<0.0001; K i: RS =0.429, P=0.0005) and lowest for areal BMD (SUV: RS =0.353, P=0.005; K i: RS =0.274, P=0.03). The results demonstrate significant positive correlations between SUV and K i and vBMD measurements in the form of HU from CT or BMAD and areal BMD from DXA. These findings justify further exploration of the relationship between SUV and K i [18F]NaF PET/CT measurements and CT or DXA measurements of vBMD to examine whether normalization for bone density might improve their correlation with bone metabolic activity as measured by bone biopsy.


Galectin-3 Expression in Pancreatic Cell Lines Under Distinct Autophagy-Inducing Stimulus.
Hypoxia and nutrient deprivation are responsible for inducing malignant behavior in neoplastic cells. In these conditions, metabolic stress leads the cells to enhance their autophagic flux and to activate key molecules for homeostasis maintenance. Galectin-3 (Gal-3) is upregulated in pancreatic cancer and it is activated under the hypoxic atmosphere. We aimed to analyze the most effective autophagic-inducing conditions in pancreatic ductal adenocarcinoma cells and the effect exerted under these conditions in association with hypoxia on the Gal-3 expression. Gal-3 and the microtubule-associated protein light chain 3 beta (LC3) were accessed through western blot and immunofluorescence. Degradative vacuole quantification was analyzed by transmission electronic microscopy, and inhibition of Gal-3 was performed using siRNA. According to the analyses, the most effective conditions in the inducement of autophagy for PANC-1 and MIA PaCa-2 cells were nutritional deprivation and complete amino acid/glucose deprivation, respectively. PANC-1 cells presented higher Gal-3 when they were submitted to 24 h of nutritional deprivation alone and simultaneously nutritional and oxygen deprivation. Inhibition of Gal-3 causes a decrease of LC3 levels in all experimental conditions. These results confirm that Gal-3 is modulated by microenvironment factors and the possibility of Gal-3 participating in an adaptive response from PDAC cells to extreme conditions.


Feeding strategies for managing heat load in feedlot cattle.
Eighty-four Bos taurus crossbred steers were used to investigate effects of level and duration of limit-feeding feedlot cattle in a hot environment. Pens (four/treatment) of steers (seven/pen) were fed feedlot finishing diets and randomly assigned to the following treatments: 1) restricted to approximately 75% of feed consumed when offered ad libitum for 21-d duration (RES21); 2) restricted to approximately 75% of ad libitum for 42-d duration (RES42); and 3) feed offered ad libitum (ADLIB). Tympanic temperatures (TT) were measured via thermistors placed in the ear canal and attached to data loggers. Restricting feed intake for both 21- and 42-d reduced tympanic temperature when compared with ADLIB treatment groups under hot environmental conditions. Temperature reductions exceeded 0.5 degrees C (P < 0.05) depending on time of day. The reduced tympanic temperature is likely due to a reduction in metabolic heat load and/or a concurrent reduction in metabolic rate. Within respective periods, no differences (P > 0.05) were found among treatments for panting or bunching score. However, different proportions of cattle were found to be bunching and panting with ADLIB cattle displaying a greater number of bunched steers that were panting when compared with the other groups. When averaged across diet treatments, dark-colored cattle had the greatest percentage of cattle showing moderate to excessive panting, while light-colored cattle displayed the least panting under thermoneutral climatic conditions. Under hot (mean daily temperature-humidity index >74) conditions, dark-colored cattle tended to bunch more (P = 0.073) and pant more (P < 0.01) than light-colored cattle. Mean TT were 0.2 to 0.6 degrees C (P < 0.05) greater for dark- vs light-colored cattle under hot conditions. Limit-feeding feedlot cattle during early summer is a successful tool for enhancing animal comfort by alleviating the combined effects of high climatic and metabolic heat load.


[The analysis of failure diode laser treatment of active phase of retinopathy of prematurity].
The aim of this study was to analyze risk factors influencing diode laser treatment of active phase of retinopathy of prematurity. 106 premature infants were treated with the use of diode laser at Department of Neonatology since 1.01.2003-30.11.2004. The mean gestational age was 27.3, mean birth weight 1022 g. The earliest age of laser therapy was 37th day of life and the latest one 104th day (mean 64.3 days). 18 neonates were excluded from the study because they were from other departments, and 2 because they died. The results of treatment of the rest 86 children were evaluated. The occurence of retinal detachment or macular ectopy was classified as unfavorable outcome. The complete remission was obtained in 143 eyes (83.2%),the macular ectopy was diagnosed in 10 eyes (5.8%), the retinal detachment in 19 eyes (11%). Children with unfavorable outcomes were divided in two groups. The first one included 6 extremely immature infants, born between 24 to 25th week of gestation. The ROP in this group was diagnosed in I zone with plus disease. The second group included children with gestation age from 28 to 32 Hbd. 9 of them had severe infection (sepsis, congenital pneumonia, urinary tract infection), 1 serious maternal history (R type of diabetes). The procedure of laser therapy was performed on average at 72th day of life at the first group, and at 50th day at the second one. (1) Important risk factors of treatment failure of active phase of retinopathy of prematurity, is low gestational age and infections which take place in perinatal period. (2) In the second group of infants (gestational age from 28-32 Hbd), with treatment failure the retinopathy of prematurity occured much more earlier then in the group with successful treatment. (3) When making decision of the patient treatment besides estimation of the eye fundus and the progress of changes, we should also consider risk factors which have influence on retinopathy of prematurity, after treatment.


Direct and specific chemical control of eukaryotic translation with a synthetic RNA-protein interaction.
Sequence-specific RNA-protein interactions, though commonly used in biological systems to regulate translation, are challenging to selectively modulate. Here, we demonstrate the use of a chemically-inducible RNA-protein interaction to regulate eukaryotic translation. By genetically encoding Tet Repressor protein (TetR)-binding RNA elements into the 5'-untranslated region (5'-UTR) of an mRNA, translation of a downstream coding sequence is directly controlled by TetR and tetracycline analogs. In endogenous and synthetic 5'-UTR contexts, this system efficiently regulates the expression of multiple target genes, and is sufficiently stringent to distinguish functional from non-functional RNA-TetR interactions. Using a reverse TetR variant, we illustrate the potential for expanding the regulatory properties of the system through protein engineering strategies.


Hydrolysis behavior of bamboo fiber in formic acid reaction system.
The process of conversion of bamboo fiber into fermentable glucose in the formic acid reaction system was investigated using cross-polarization/magic angle spinning (13)C-nuclear magnetic resonance (CP/MAS (13)C NMR), X-ray diffraction. The results indicated that formic acid as an active agent was able to effectively penetrate into the interior space of the cellulose molecules, thus collapsing the rigid crystalline structure and allowing hydrolysis to occur easily in the amorphous zone as well as in the crystalline zone. The bamboo fiber was hydrolyzed using formic acid and 4% hydrochloric acid under mild conditions. The effects of temperature (55-75 degrees C), retention time (0-9 h) and catalyst, the concentration of glucose and the degradation pathway of glucose were analyzed. The main degradation pathway of glucose is that the hydroxyl group on the 2-carbon is protonated and cleaved off. Aluminum iso-propoxide prevented the degradation of glucose, and the acetone promoted the degradation of glucose to levulinic acid.


Analysis of human acellular nerve allograft reconstruction of 64 injured nerves in the hand and upper extremity: a 3 year follow-up study.
Human acellular nerve allografts have been increasingly applied in clinical practice. This study was undertaken to investigate the functional outcomes of nerve allograft reconstruction for nerve defects in the upper extremity. A total of 64 patients from 13 hospitals were available for this follow-up study after nerve repair using human acellular nerve allografts. Sensory and motor recovery was examined according to the international standards for motor and sensory nerve recovery. Subgroup analysis and logistic regression analysis were conducted to identify the relationship between the known factors and the outcomes of nerve repair. Mean follow-up time was 355 ± 158 (35-819) days; mean age was 35 ± 11 (14-68) years; average nerve gap length was 27 ± 13 (10-60) mm; no signs of infection, tissue rejection or extrusion were observed among the patients; 48/64 (75%) repaired nerves experienced meaningful recovery. Univariate analysis showed that site and gap length significantly influenced prognosis after nerve repair using nerve grafts. Delay had a marginally significant relationship with the outcome. A multivariate logistic regression model revealed that gap length was an independent predictor of nerve repair using human acellular nerve allografts. The results indicated that the human acellular nerve allograft facilitated safe and effective nerve reconstruction for nerve gaps 10-60 mm in length in the hand and upper extremity. Factors such as site and gap length had a statistically significant influence on the outcomes of nerve allograft reconstruction. Gap length was an independent predictor of nerve repair using human acellular nerve allografts. Copyright © 2016 John Wiley & Sons, Ltd.


Hormone therapy effects on social behavior and activity levels of surgically postmenopausal cynomolgus monkeys.
The purpose of the experiments reported here was to investigate central nervous system effects of commonly prescribed postmenopausal hormone therapies in a primate model, the cynomolgus monkey (Macaca fascicularis). The results of two experiments are reported. In the first, ovariectomized adult cynomolgus monkeys were treated for eight weeks each with oral micronized 17beta-estradiol (E2) (n=23), E2+medroxyprogesterone acetate (MPA) (n=23), E2+progesterone (P4) (n=23), and placebo (n=23) using a crossover design. In the second, ovariectomized adult cynomolgus monkeys were treated for eight weeks with oral micronized E2+oral micronized P4 (n=10), or E2+intravaginal micronized P4 delivered via a Silastic ring (n=8), or oral placebo and intravaginal placebo (n=5), using a parallel arm design. Behavior was recorded during weeks two through four. Cerebrospinal fluid (CSF) and blood were sampled, and 24h heart rate recorded by telemetry during weeks five through seven. Monoaminergic metabolites were assayed in CSF, and cortisol was assayed in serum. There were no significant effects of treatment on CSF monoaminergic metabolites or heart rate. E2+MPA increased cortisol concentrations. While there were some differences in effects between experiments, both progestogens and both routes of administration increased time spent resting, particularly resting in body contact, resulting in increased passive affiliative interaction. Thus, synthetic progestogens appear to be as sedating as progesterone, and the ring delivery system does not appear to protect the central nervous system from effects of progestogens. Further research is needed to explore social context as an important feature of behavioral response to steroid hormone regimens and to verify and extend knowledge of systemic effects of vaginal ring-delivered progestogens.


"Devil's gardens" in the Ecuadorian Amazon--association of the allelopathic tree Duroia hirsuta (Rubiaceae) and its "gentle" ants.
The relationship between the myrmecophytic tree Duroia hirsuta (Rubiaceae) and its associated, minute, non-aggressive Myrmelachista sp. (Formicinae) ants is poorly investigated, even though the small understory tree is surrounded by a conspicuos zone almost devoid of vegetation. As to the cause of these barely vegetated "Devil's Gardens", two alternative interpretations exist in the literature: the pruning activity of ants and allelopathy. We investigated the mutualistic Duroia-Myrmelachista system in the Ecuadorian Amazon, focusing on the phenomenon of the areas of reduced vegetation, as well as on the natural history of the associated ants. We conducted planting experiments and behavioral studies, including coccoids, the third partner in this mutualism. Because the well-studied Triplaris americana (Polygonaceae) possesses analogous vegetation free surroundings created by the assiduous mechanical pruning of its mutualistic ant partner (Pseudomyrmex sp.), parallel comparative investigations of this ant-plant association were conducted. The two systems are not equivalent; it was demonstrated that the remarkably inactive Myrmelachista ants are not responsible for the bare areas around Duroia plants. Instead, we regard allelopathy as the causative agent for the clear zones.


Response time studies of a new, portable mass spectrometer.
Mass spectrometers are frequently used by anesthesiologists perioperatively to monitor patients' respiratory function and levels of inhaled anesthetics. Due to size, complexity and expense, they are typically used in a time-sharing manner which degrades their performance. We assessed the accuracy of the Random Access Mass Spectrometer (RAMS), Marquette Electronics) which is small enough to be dedicated to a single patient. We compared the 10-90% rise times for O2, CO2, N2O and isoflurane for the RAMS with different catheter configurations to those of a MedSpect mass spectrometer (Allegheny International Medical Technology) operating under ideal conditions. For CO2 the lag of the RAMS relative to the MedSpect was also measured. Next, perioperative conditions were stimulated by ventilating anesthetized dogs with a variety of inhalatory gases and ventilatory parameters, and the interchangeability of the two devices was assessed. When fitted with a catheter with minimal dead space the MedSpect had rise times of 0.11-0.12 sec while the RAMS had rise times of 0.07-0.12 sec and a delay of 0.19 sec compared to the MedSpect. The rise times and delay of the RAMS increased when using a larger catheter and water trap. Although there were statistically significant differences in some values for inhaled and end-tidal gases under simulated perioperative conditions, particularly at the higher frequencies, these differences were small and for most purposes not clinically significant. Our results demonstrate that the RAMS configured for clinical conditions performs nearly as well as the MedSpect under ideal conditions. The small differences between the two, confined almost entirely to their end-tidal CO2 values, could be due to differences in instrument calibration, by the larger sampling catheter commonly used in clinical settings, or by a combination of both factors. Therefore the RAMS is sufficiently accurate for clinical use and would alleviate problems associated with time-shared mass spectrometers.


Characterization of essential domains and plasticity of the classical Swine Fever virus Core protein.
Pestiviruses are pathogens of cloven-hoofed animals, belonging to the Flaviviridae. The pestiviral particle consists of a lipid membrane containing the three envelope glycoproteins Erns, E1, and E2 and a nucleocapsid of unknown symmetry, which is composed of the Core protein and the viral positive-sense RNA genome. The positively charged pestiviral Core protein consists of 86 to 89 amino acids. To analyze the organization of essential domains, N- and C-terminal truncations, as well as internal deletions, were introduced into the Core coding sequence in the context of an infectious cDNA clone of classical swine fever virus strain Alfort. Amino acids 179 to 180, 194 to 198, and 208 to 212 proved to be of special importance for the generation of progeny virus. The results of transcomplementation of a series of C-terminally truncated Core molecules indicate the importance of Ala255 at the C terminus. The plasticity of Core protein was examined by the construction of concatemeric arrays of Core coding regions and the insertion of up to three yellow fluorescent protein (YFP) genes between two Core genes. Even a Core fusion protein with more than 10-fold-increased molecular mass was integrated into the viral particle and supported the production of infectious progeny virus. The unexpected plasticity of Core protein brings into question the formation of a regular icosahedric particle and supports the idea of a histone-like protein-RNA interaction. All viruses with a duplicated Core gene were unstable and reverted to the wild-type sequence. Interestingly, a nonviable YFP-Core construct was rescued by a mutation within the C-terminal domain of the nonstructural protein NS3.


The effects of different irrigation solutions on the bond strength of cemented fiber posts.
To study effect of root canal irrigant solutions on the bond strength of cemented fiber posts, and resin cement-tags in root canal dentin. Fifty-two human single-rooted anterior teeth were selected and stored in 1% sodium azide. Crowns were sectioned 2 mm incisal to the cemento enamel junction with diamond bur at high speed under water-cooling. After standard root canal treatment to tested teeth, they were randomly assigned into four groups (n=13) corresponding to the endodontic irrigant solution that was used during post space preparation. The treatment groups were: Group 1: 6.15% sodium hypochlorite; Group 2: 17% EDTA; Group 3: 6.15% NaOCl +17% EDTA; Group 4: 6.15% NaOCl +0.12% chlorhexidine solution. Parallel-sided fiber posts were used for all specimens. Samples were embedded in a cylindrical PVC mounting jig to facilitate perpendicular sectioning. With low speed diamond saw, cervical and apical specimens from each tooth were obtained and subjected to push-out test using universal testing machine. SEM was used to examine the root-canal dentin surface. Statistical analysis revealed significant difference among the groups at P<0.05. Group 2 treated with EDTA, had the highest bond strength; 18.63±2.85 MPa in cervical specimens and 13.49±3.67 MPa in apical specimens. The cervical specimens of Groups 2 and 3 were significantly different than the apical specimens in the same group. Adhesive failure between cement and dentin was the main failure mode observed in all groups. Groups irrigated with EDTA showed cleaner dentin surface and better resin tag formation, while groups treated with NaOCl showed less resin tag formation and insufficient smear layer removal. NaOCl had adverse effect on bond strength of self-adhesive resin cement to radicular dentin. EDTA irrigant solution produced higher bond strength and was more effective in removing smear layer than NaOCl.


Influence of oceanic factors on Anguilla anguilla (L.) over the twentieth century in coastal habitats of the Skagerrak, southern Norway.
The European eel (Anguilla anguilla L.) is distributed in coastal and inland habitats all over Europe, but spawns in the Sargasso Sea and is thus affected by both continental and oceanic factors. Since the 1980s a steady decline has been observed in the recruitment of glass eels to freshwater and in total eel landings. The eel is considered as critically endangered on the International Union for the Conservation of Nature and Natural Resources Red List of species. The Skagerrak beach seine survey from Norway constitutes the longest fishery-independent dataset on yellow/silver eels (starting in 1904). The Skagerrak coastal region receives larvae born in the Sargasso Sea spawning areas that have followed the Gulf Stream/North Atlantic Drift before they penetrate far into the North Sea. The Skagerrak coastal time series is therefore particularly valuable for exploring the impacts of oceanic factors on fluctuations in eel recruitment abundance. Analyses showed that Sargasso Sea surface temperature was negatively correlated with eel abundance, with a lag of 12 years revealing a cyclic and detrimental effect of high temperatures on the newly hatched larvae. The North Atlantic Oscillation index and inflow of North Atlantic water into the North Sea were negatively correlated with eel abundance, with a lag of 11 years. Increased currents towards the North Atlantic during high North Atlantic Oscillation years may send larvae into the subpolar gyre before they are ready to metamorphose and settle, resulting in low recruitment in the northern part of the distribution area for these years. The Skagerrak time series was compared with glass eel recruitment to freshwater in the Netherlands (Den Oever glass eel time series), and similar patterns were found revealing a cycle linked to changes in oceanic factors affecting glass eel recruitment. The recent decline of eels in the Skagerrak also coincided with previously documented shifts in environmental conditions of the North Sea ecosystem.


Sport, physical activity and social exclusion.
This paper is a brief review of the concept of social exclusion and its evolution. I address which individuals are excluded from sport and physical activity and how; link inclusion policies to the 'cross-cutting issues' and the idea of social capital; and outline the intervention policies being adopted in the new sport strategy 'Game Plan' (). I address the link between transport, exercise and health in a case study. Since these policies are new, research and evaluation has been short term and scattered, and outcome measurements have not yet received academic or professional consensus, it is too soon to say for sure what works or even to confirm what is best practice.


Dietary Intake and Type 2 Diabetes.
This editorial aims to examine the risk factors associated with type 2 diabetes and to discuss the evidence relating to dietary strategies for managing people with this condition. It is clear from the evidence presented that a range of dietary interventions can provide useful approaches for managing people with type 2 diabetes, including the regulation of blood glucose and lipid parameters, and for reducing the risks of acute and chronic diabetic complications.


[Alexithymia impact on EEG activity of anterior and posterior regions of the right hemisphere while experiencing positive and negative emotions].
The present study was aimed to assess the influence of alexithymia (an emotional disorder associated with difficulties in identification and describing personal feelings) on regional activity while experiencing positive or negative emotional activation. The 62-channels EEG was recorded in non-alexithymic (n = 27) and alexithymic (n = 17) subjects viewing emotional film clips. Effects of alexithymia were found in the upper theta, low and upper alpha frequency bands; for all the bands the alexithymics exhibited greater reactivity of the right hemisphere, suggesting enhanced avoidance motivational tendencies, negative effect and autonomic arousal. The present phenomenon suggests engagement of additional cognitive resources in identification of emotional feelings. Such a 'cognitive effort' can be a result of impaired referential links and deficit of symbolic processing of emotions.


Recent Update in the Diagnosis and Treatment of Bone Frailty in Patients with a Distal Radius Fracture.
Distal radius fracture (DRF) is the most common upper extremity fracture in the elderly. Patients with a DRF have a two to fourfold higher risk of a subsequent fracture than those with no history of fractures, and DRFs occur on average 15 years earlier than hip fractures. Therefore, patients with a DRF offer physicians an important opportunity to diagnose and treat osteoporosis to prevent a secondary fracture. In this review, we provide recent update in the diagnosis and treatment of bone frailty in patients with a DRF.


The efficiency of autologous stem cell transplantation as the first-line treatment for nodal peripheral T-cell lymphoma: results of a systematic review and meta-analysis.
Nodal peripheral T cell lymphoma (PTCL) confers a dismal prognosis when treated with conventional chemotherapy. Autologous stem cell transplantation (ASCT) seems a reasonable alternative in eligible patients. Nevertheless, a consensus on the role of ASCT as the first-line consolidation therapy for nodal PTCL patients has not been reached so far. A quantitative meta-analysis was performed via a systematic search in PubMed, Web of Science, Embase, and The Cochrane Library. The overall survival (OS), progression-free survival (PFS), hazard ratio (HR), and 95% confidence intervals (CIs) were compared and calculated from database inception to September 2021. Twelve articles were eligible. The results showed that ASCT could improve the survival of patients compared with chemotherapy alone. In terms of subtype analysis, results showed that angioimmunoblastic T cell lymphoma (AITL) patients could benefit more from chemotherapy followed by ASCT. Statistical differences were also confirmed for OS and PFS in different remission status, clinical stage, performance status (PS), chemotherapy regimen, and gender. ASCT could serve as the first-line consolidation treatment strategy for nodal PTCL patients, especially AITL patients. Early clinical stage, good PS status, CR before transplantation, CHOEP regimen, and female patients may indicate a better outcome.


Absorbed radiation dose to humans from technetium-99m-teboroxime.
Tissue distribution data obtained in nine normal volunteers were used to estimate the radiation dose to humans after intravenous administration of 99mTc-teboroxime (Cardiotec). Organ uptake as percent of injected dose was measured using quantitative SPECT. Non-linear regression analysis was performed on the organ time-activity data using SYSTAT software. Cumulative activities in these organs were determined by calculating the area under the respective curves after accounting for the physical decay of the radionuclide. The absorbed dose to individual organs was estimated using the MIRDOSE 2 program. The gallbladder and the upper large intestine (ULI) are the target organs and will receive respectively 26.5 and 33.2 muGy/MBq (98 and 123 mrad/mCi) 99mTc-teboroxime under the assumption that the gallbladder empties every 6 hr. The dose to the gallbladder decreases at shorter emptying intervals; with intervals of 3, 4, and 5 hr, the respective doses to the gallbladder are 18.2, 21.0 and 23.7 muGy/MBq (67.4, 77.8, and 87.9 mrad/mCi) 99mTc-teboroxime. However, the dose to ULI remains almost constant at 123 mrad/mCi and will be the limiting factor.


The relationship of the peripheral leukocyte count and cigarette smoking to pulmonary function among adult men.
The relationship of the peripheral leukocyte count to pulmonary function was examined in 1,510 men. There were 466 current cigarette smokers, 485 former cigarette smokers, and 559 never smokers enrolled in a longitudinal study of aging. Spirometry was performed twice on all subjects ten years apart. A multiple regression analysis indicated that leukocyte count was inversely related to baseline levels of both forced vital capacity (FVC) (p less than 0.001) and forced expiratory volume in one second (FEV1) (p less than 0.001) after adjustment for age, height, and smoking habits. Similarly, change in leukocyte count over a ten-year period was inversely related to follow-up levels of both FVC (p = 0.036) and FEV1 (p = 0.004) after adjustment for baseline levels of leukocyte count, pulmonary function, age, height, and smoking habits. These data suggest that the peripheral leukocyte count is an important determinant of level of pulmonary function. Further investigation of the pathophysiologic mechanisms involved in this association seems indicated.


Analysis of chromosome and karyotype in Bali cattle and Simmental-Bali (Simbal) crossbreed cattle.
Chromosome analysis is very important part for the initial analysis of genetics. Some genetics abnormalities can be detected at the chromosome level and are usually associated with the inherited diseases. Accurate and prompt information need to be obtained for the purposes of the prevention of the genetics abnormalities genetics risk. This study was aimed to find out and to analyze the different size and morphological chromosome of Bali cattle and its crosses with Simmental cattle (Simbal cattle). Ten Simbal cattle (5 male and 5 female) and 5 female Bali cattle were used in this study. Five milliliter blood was collected using venous puncture through the jugular vein of each cattle. Chromosome was derived from white blood cells (lymphocyte) of peripheral blood. The Bali cattle and Simbal cattle have diploid chromosome (2n) of 60, with 29 pair of autosomes and one pair of sex chromosome. All autosomes are acrocentric with centromere index of 25.13 to 29.52% for Simbal cattle and 21.61 to 24.84% for Bali cattle. Sex chromosomes ware sub-metacentric in Simbal cattle either male or female and metacentric in female Bali cattle. Average length of chromosomes of female Simbal, male Simbal and female Bali cattle were 0.29 +/- 0.04 micron, 0.30 +/- 0.05 micron and 0.24 +/- 0.02 micron, respectively. Chromosome size of female Bali cattle was smaller than Simbal cattle.


Predictivity of thyroid autoantibodies for the development of thyroid disorders in children and adolescents with Type 1 diabetes.
To investigate the prevalence of thyroid autoantibodies and their significance for the development of thyroid disorders in children and adolescents with Type 1 diabetes. Antibodies to thyroglobulin (anti-TG) and thyroperoxidase (anti-TPO) were measured in 216 patients (113 boys; median age 12.9 years (range 1-22 years)) with Type 1 diabetes (diabetes duration 2.5 years (0-14 years)) in a cross-sectional study. Sixteen patients with significantly elevated anti-TPO titres were followed longitudinally (6.0 years (4-13 years)) including the measurement of anti-TPO, anti-TG, T(3), T(4), thyroid-stimulating hormone (TSH) and ultrasound assessment. Twenty-two patients (10.0%) had significantly elevated titres of anti-TPO, 19 (8.7%) of anti-TG and 13 (5.9%) of both autoantibodies. Girls had more frequently elevated anti-TPO antibodies than boys (P < 0.05). Eight of 16 patients (50%) developed thyroid disorders defined by a TSH elevation (> or = 4.5 microU/ml) and/or sonographic thyroid abnormalities during a median time of 3.5 years (2-6 years) after first detection of anti-TPO positivity. They were characterized by higher levels of anti-TPO (P = 0.001) and a more frequent coexistence of anti-TG antibodies (P = 0.002) than those with no development of thyroid disorder even after an observation period of 5.5 years (5-10 years). Because 50% of children with diabetes and significant titres of anti-TPO develop thyroid problems within 3-4 years, examinations of thyroid antibodies should be performed yearly. In cases of significant antibody titres, thyroid function tests and ultrasound assessment are recommended in order to minimize the risk of undiagnosed hypothyroidism in these patients.


Ultrastructural study of the early development and localization of Loma salmonae in the gills of experimentally infected rainbow trout.
The early ultrastructural stages of Loma salmonae were studied in the gills of experimentally infected rainbow trout. No parasitic stages were identified during the first 2 wk of the infection. By week 3 postexposure (PE), uninucleate and binucleate meronts were recognized within host cells (no xenomas) associated with the capillary channels of secondary lamellae and lamellar arteries. An inflammatory reaction was absent. In secondary lamellae, infected cells were isolated from the capillary lumen, and some were recognized as pillar cells. In lamellar arteries, infected cells were localized beneath the endothelium and not in the lumen. Inflammatory reaction and destruction of parasites inside blood cells in the lumen of secondary lamellae were observed by week 4 PE. Three hypotheses, i.e., isolation, internalization, and evasion, are proposed to explain the localization of the infected cells in the gills. It is concluded that meronts are the earliest parasitic stage observed by week 3 PE, pillar cells are secondarily infected by phagocytosis of infected cells in the blood, endothelial cells of gills are not infected, and inflammatory response to the parasite starts by week 4 PE.


Effect of surface etching on the efficiency of ZnO-based dye-sensitized solar cells.
In this study, we report on an insight into the influential factors of the conversion efficiency of ZnO-based dye-sensitized solar cells. With ZnO nanoflowers as the photoelectric anode and dye N719 as the sensitizer, we found with the addition of N719 that the surfaces of ZnO nanoflowers were slightly etched at the beginning and gradually destroyed with time. On the basis of these observations, a series of experiments were further carried out to distinguish the influence of dye-induced aggregation from that of dye-induced etching on the solar-to-electric conversion efficiency. SEM observation reveals that there were no obvious dye/Zn(2+) aggregations on any of the samples. XRD results indicate that there was no new phase formed during the dye-sensitizing process. I-V measurements reveal clearly that the efficiency of ZnO-based DSSCs was inversely proportional to the etching level of ZnO surfaces. We concluded that the dye-induced etching of the ZnO anode may be an assignable cause that results in the low efficiency of ZnO-based DSSCs. The etching of ZnO may lead to low surface absorption efficiency of the dye, low electron mobility, and a high surface recombination ratio of photocarriers. Therefore, we suggest that special attention should be paid to protecting the surface structure of the ZnO anode during the dye-sensitizing process.


Dual Role of SIRT1 in Autophagy and Lipid Metabolism Regulation in Osteoarthritic Chondrocytes.
Background and Objectives: Osteoarthritis (OA) is one of the most common and highly prevalent types of arthritis, also considered a multiphenotypic disease with a strong metabolic component. Ageing is the primary risk factor for OA, while the age-related decline in autophagic activity affects cell function and chondrocyte homeostasis. The aim of this study was to investigate the role of sirtuin 1 (SIRT1) in autophagy dysregulation and lipid metabolism in human OA chondrocytes. Materials and Methods: OA chondrocytes were treated with Resveratrol, Hydroxycloroquine (HCQ) or 3-Methyladenine (3-MA) and HCQ or 3-MA followed by siRNA against SIRT1 (siSIRT1). Then, SIRT1, AcNF-κBp65, LOX-1 and autophagy-related proteins ATG5, ATG13, PI3K class III, Beclin-1, LC3 and ULK protein levels were evaluated using Western blot. Normal articular chondrocytes were treated under serum starvation and/or siSIRT1, and the protein expression levels of the above autophagy-related proteins were evaluated. The staining patterns of LC3/p62 and LOX-1 were analyzed microscopically by immunofluorescence. SIRT1/LC3 complex formation was analyzed by immunoprecipitation. Results: SIRT1 and LOX-1 protein expression were negatively correlated in OA chondrocytes. SIRT1 regulated LOX-1 expression via NF-κΒ deacetylation, while treatment with Resveratrol enhanced SIRT1 enzymatic activity, resulting in LOX-1 downregulation and autophagy induction. In OA chondrocytes, SIRT1 was recognized as an autophagy substrate, formed a complex with LC3 and was consequently subjected to cytoplasmic autophagosome-lysosome degradation. Moreover, siSIRT1-treated normal chondrocytes showed decreased autophagic activity, while double-treated (siSIRT1 and serum starvation) cells showed no induction of autophagy. Conclusions: Our results suggest that SIRT1 regulates lipid homeostasis through LOX-1 expression regulation. Additionally, we indicate that the necessity of SIRT1 for autophagy induction in normal chondrocytes, together with its selective autophagic degradation in OA chondrocytes, could contribute to autophagy dysregulation in OA. We, therefore, suggest a novel regulatory scheme that functionally connects lipid metabolism and autophagy in late-stage OA.


Interpretation of size-exclusion chromatography for the determination of molecular-size distribution of human immunoglobulins.
Molecular-size distribution by size-exclusion chromatography (SEC) [1] is used for the quantification of unwanted aggregated forms in therapeutic polyclonal antibodies, referred to as human immunoglobulins (Ig) in the European Pharmacopoeia. Considering not only the requirements of the monographs for human normal Ig (0338, 0918 and 2788) [2-4], but also the general chapter on chromatographic techniques (2.2.46) [5], several chromatographic column types are allowed for performing this test. Although the EDQM knowledge database gives only 2 examples of suitable columns as a guide for the user, these monographs permit the use of columns with different lengths and diameters, and do not prescribe either particle size or pore size, which are considered key characteristics of SEC columns. Therefore, the columns used may differ significantly from each other with regard to peak resolution, potentially resulting in ambiguous peak identity assignment. In some cases, this may even lead to situations where the manufacturer and the Official Medicines Control Laboratory (OMCL) in charge of Official Control Authority Batch Release (OCABR) have differing molecular-size distribution profiles for aggregates of the same batch of Ig, even though both laboratories follow the requirements of the relevant monograph. In the present study, several formally acceptable columns and the peak integration results obtained therewith were compared. A standard size-exclusion column with a length of 60 cm and a particle size of 10 µm typically detects only 3 Ig fractions, namely monomers, dimers and polymers. This column type was among the first reliable HPLC columns on the market for this test and very rapidly became the standard for many pharmaceutical manufacturers and OMCLs for batch release testing. Consequently, the distribution of monomers, dimers and polymers was established as the basis for the interpretation of the results of the molecular-size distribution test in the relevant monographs. However, modern columns with a smaller particle size provide better resolution and also reveal a class of components designated here as oligomers. This publication addresses the interpretation of the SEC test for Ig with respect to the following questions: - how can molecular-size distribution tests benefit from the use of the most recent column technology without changing the sense of well-established quality parameters? - is it possible to mathematically define a way to interpret chromatograms generated with various column types with the same fractionation range but different resolution power? - how should oligomers be considered regarding compliance with compendial specifications?


Phosphopeptide enrichment with TiO2-modified membranes and investigation of tau protein phosphorylation.
Selective enrichment of phosphopeptides prior to their analysis by mass spectrometry (MS) is vital for identifying protein phosphorylation sites involved in cellular regulation. This study describes modification of porous nylon substrates with TiO2 nanoparticles to create membranes that rapidly enrich phosphopeptides. Membranes with a 22-mm diameter bind 540 nmol of phosphoangiotensin and recover 70% of the phosphopeptides in mixtures with a 15-fold excess of nonphosphorylated proteins. Recovery is 90% for a pure phosphopeptide. Insertion of small membrane disks into HPLC fittings allows rapid enrichment from 5 mL of 1 fmol/μL phosphoprotein digests and concentration into small-volume (tens of microliters) eluates. The combination of membrane enrichment with tandem mass spectrometry reveals seven phosphorylation sites from in vivo phosphorylated tau (p-tau) protein, which is associated with Alzheimer's disease.


Design of Efficient Catalysts with Double Transition Metal Atoms on C2N Layer.
Heterogeneous catalysis often involves molecular adsorptions to charged catalyst site and reactions triggered by catalyst charges. Here we use first-principles simulations to design oxygen reduction reaction (ORR) catalyst based on double transition metal (TM) atoms stably supported by 2D crystal C2N. It not only holds characters of low cost and high durability but also effectively accumulates surface polarization charges on TMs and later deliveries to adsorbed O2 molecule. The Co-Co, Ni-Ni, and Cu-Cu catalysts exhibit high adsorption energies and extremely low dissociation barriers for O2, as compared with their single-atom counterparts. Co-Co on C2N presents less than half the value of the reaction barrier of bulk Pt catalysts in the ORR rate-determining steps. These catalytic improvements are well explained by the dependences of charge polarization on various systems, which opens up a new strategy for optimizing TM catalytic performance with the least metal atoms on porous low-dimensional materials.


The GTPase activating factor for transducin in rod photoreceptors is the complex between RGS9 and type 5 G protein beta subunit.
Proteins of the regulators of G protein signaling (RGS) family modulate the duration of intracellular signaling by stimulating the GTPase activity of G protein alpha subunits. It has been established that the ninth member of the RGS family (RGS9) participates in accelerating the GTPase activity of the photoreceptor-specific G protein, transducin. This process is essential for timely inactivation of the phototransduction cascade during the recovery from a photoresponse. Here we report that functionally active RGS9 from vertebrate photoreceptors exists as a tight complex with the long splice variant of the G protein beta subunit (Gbeta5L). RGS9 and Gbeta5L also form a complex when coexpressed in cell culture. Our data are consistent with the recent observation that several RGS proteins, including RGS9, contain G protein gamma-subunit like domain that can mediate their association with Gbeta5 (Snow, B. E., Krumins, A. M., Brothers, G. M., Lee, S. F., Wall, M. A., Chung, S., Mangion, J., Arya, S., Gilman, A. G. & Siderovski, D. P. (1998) Proc. Natl. Acad. Sci. USA 95, 13307-13312). We report an example of such a complex whose cellular localization and function are clearly defined.


Quantitative fluorescence measurement of cardiac Na+/Ca2+ exchanger inhibition by kinetic analysis in stably transfected HEK293 cells.
We developed a method to quantitatively evaluate the potency of Na+/Ca2+ exchanger (NCX) inhibitors with fluorescence microscopy in NCX1-transfected HEK 293 cells. The reverse mode and forward mode NCX activities were measured as the ascending slope of the early phase increase in cytoplasmic Ca2+ concentration after change to low Na+ extracellular solution and the descending rate (inverse of the exponential time constant) on return to normal solution, respectively. Both modes of NCX were inhibited by SEA0400 (2-[4-[(2,5-difluorophenyl)methoxy]phenoxy]-5-ethoxyaniline) and KB-R7943 (2-[2-[4-(4-nitrobenzyloxy)phenyl]ethyl]isothiourea methanesulfonate), and the concentration-inhibition relationships for both inhibitors were in good agreement with those previously reported in voltage clamped cardiomyocytes.


[Evaluation of autonomy and disability in elderly people in France, using O.E.C.D. questions and the number of illnesses (author's transl)].
Three approaches (traditional nosology, functional ability, socio-economic implications) to health status have been taken into account simultaneously in the development of different indicators from a sample of 2802 elderly persons aged 65 years and over: -using traditional nosology: the existence of different illnesses and total number of conditions for an individual, -on the functional level: the level of disability taking into account both physical and mental deficiencies, -on the socio-economic level: the basic level of dependency established according to the person's ability to perform activities of daily living. The techniques discussed will be generalized to larger populations allowing comparisons between the elderly living at home or in a private institution, and between socio-demographic groups.


Stability of complex coacervate core micelles containing metal coordination polymer.
We report on the stability of complex coacervate core micelles, i.e., C3Ms (or PIC, BIC micelles), containing metal coordination polymers. In aqueous solutions these micelles are formed between charged-neutral diblock copolymers and oppositely charged coordination polymers formed from metal ions and bisligand molecules. The influence of added salt, polymer concentration, and charge composition was investigated by using light scattering and cryo-TEM techniques. The scattering intensity decreases strongly with increasing salt concentration until a critical salt concentration beyond which no micelles exist. The critical micelle concentration increases almost exponentially with the salt concentration. From the scattering results it follows that the aggregation number decreases with the square root of the salt concentration, but the hydrodynamic radius remains constant or increases slightly. It was concluded that the density of the core decreases with increasing ionic strength. This is in agreement with theoretical predictions and is also confirmed by cryo-TEM measurements. A complete composition diagram was constructed based on the composition boundaries obtained from light scattering titrations.


Effect of time of day on the accuracy and consistency of the badminton serve.
In badminton, the short doubles serve requires accuracy, whereas the long singles serve also requires power. The present study investigated if there are time-of-day effects for these two types of serve. Eight male badminton players completed two familiarization sessions before being tested at 08:00, 14:00 and 20:00 hours, the session times being counterbalanced. At each session, subjects performed ten warm-up serves, followed by ten short and ten long test serves. A 1 m2 yellow grid was placed at the front of the recipient's service box (short serve) or at the back of the court along the centre line (long serve). Subjects scored 'hits' with their serves by putting the shuttle in the grid as close as possible to one corner of the target grids. Accuracy was assessed by calculating the average radial error (RE) of the serves, and consistency by calculating the standard deviation of the RE. Data were analysed by ANOVA. There was a time of day effect for temperature (p < 0.0005) and a trend in this for grip strength (p = 0.052). The accuracy of the 'hits' was highest at 14:00 hours (p < 0.05) and the short serve showed greater accuracy than the long serve (p < 0.0005). There was a significant effect of serve number with the short serve, subjects getting more accurate with later serves (p = 0.036). No significant variation in consistency was found (p > 0.05). In summary, the lack of agreement in profiles for temperature and the accuracies of long and short service suggests that some factor(s) that deteriorate during the course of the time spent awake might be involved, in addition to body temperature, in determining accuracy in the tasks measured.


Harnessing the power of technology for the treatment and prevention of eating disorders.
The objective of this article is to review the available literature regarding the development and evaluation of technology-enhanced interventions for eating disorders. Literature was reviewed pertaining to interventions that use technology (e.g., Internet, mobile phones) for prevention, guided self-help, treatment, relapse prevention, and carer support. A number of technology-enhanced approaches have been developed for each facet of the care spectrum, and experiences that are reported with their implementation are overall promising. However, only few of them have been evaluated in adequately designed and powered trials. This review suggests that technology-enhanced interventions offer multiple opportunities to improve care for eating disorders. More research is needed on the efficacy, effectiveness, cost-effectiveness, and reach of these approaches to ultimately estimate their public health impact. It is discussed to what extent innovative models of care integrating technology-enhanced interventions and face-to-face interventions may improve service delivery for eating disorders.


[Drug-induced acute pancreatitis: a personal contribution].
Drugs are commonly considered a rare cause of acute pancreatitis but there are an increasing number of reports of numerous medications that seem to be involved in the pathogenesis of acute pancreatitis with different degrees of causative relationship to the disease (definite--probable--possible). The number of drugs that have been associated to date with acute pancreatitis exceeds 260. The authors report here on their personal series of four cases of drug-induced acute pancreatitis (warfarin, lysinopril/hydrochlorothiazide, lamivudine/stavudine/indinavir, valproic acid), focusing on a number of epidemiological and clinical aspects.


The fur transcription regulator and fur-regulated genes in Clostridium botulinum A ATCC 3502.
Clostridium botulinum is a spore-forming bacterium that can produce a very powerful neurotoxin that causes botulism. In this study, we have investigated the Fur transcription regulators in Clostridium botulinum and Fur-regulated genes in Clostridium botulinum A ATCC 3502. We found that gene loss may be the main cause leading to the different numbers of Fur transcription regulators in different Clostridium botulinum strains. Meanwhile, 46 operons were found to be regulated by the Fur transcription regulator in Clostridium botulinum A ATCC 3502, involved in several functional classifications, including iron acquisition, iron utilization, iron transport, and transcription regulator. Under an iron-restricted medium, we experimentally found that a Fur transcription regulator (CBO1372) and two operons (DedA, CBO2610-CBO2614 and ABC transporter, CBO0845-CBO0847) are shown to be differentially expressed in Clostridium botulinum A ATCC 3502. This study has provided-us novel insights into the diversity of Fur transcription regulators in different Clostridium botulinum strains and diversity of Fur-targeted genes, as well as a better understanding of the dynamic changes in iron restriction occurring in response to this stress.


Systemic Regulatory T Cells and IL-6 as Prognostic Factors for Anatomical Improvement of Uveitic Macular Edema.
To investigate whether systemic immune mediators and circulating regulatory T cells (Tregs) could be prognostic factors for anatomic outcomes in macular edema secondary to non-infectious uveitis (UME). Multicenter, prospective, observational, 12-month follow-up study of 60 patients with UME. Macular edema was defined as central subfield thickness (CST) > 300 μm measured with spectral domain optical coherence tomography (SD-OCT). Serum samples and peripheral blood mononuclear cells (PBMC) were obtained from venous blood extraction at baseline. Serum levels of IL-1β, IL-6, IL-8, IL-17, MCP-1, TNF-α, IL-10, and VEGF were determined by Luminex. Tregs population, defined as CD3+CD4+FoxP3+ in PBMC, was determined by flow cytometry. Main outcome measure was the predictive association between searched mediators and CST sustained improvement, defined as CST < 300 microns or a 20% CST decrease, at 6 months maintained until 12-months compared to baseline levels. Multivariate logistic regression analysis showed an association between CST sustained improvement at 12 months follow-up and IL-6 and Tregs baseline levels. Higher IL-6 levels were associated with less events of UME improvement (OR: 0.67, 95% CI (0.45-1.00), P = 0.042), whereas higher levels of Tregs favored such improvement (OR: 1.25, 95% CI: 1.12-2.56, P = 0.049). Increased levels of Tregs and reduced levels of IL-6 in serum may be prognostic factors of sustained anatomical improvement in UME. These findings could enforce the opportunity to develop more efficient and personalized therapeutic approaches to improve long-term visual prognosis in patients with UME.


RNA-seq transcriptional profiling of Leishmania amazonensis reveals an arginase-dependent gene expression regulation.
Leishmania is a protozoan parasite that alternates its life cycle between the sand-fly vector and the mammalian host. This alternation involves environmental changes and leads the parasite to dynamic modifications in morphology, metabolism, cellular signaling and regulation of gene expression to allow for a rapid adaptation to new conditions. The L-arginine pathway in L. amazonensis is important during the parasite life cycle and interferes in the establishment and maintenance of the infection in mammalian macrophages. Host arginase is an immune-regulatory enzyme that can reduce the production of nitric oxide by activated macrophages, directing the availability of L-arginine to the polyamine pathway, resulting in parasite replication. In this work, we performed transcriptional profiling to identify differentially expressed genes in L. amazonensis wild-type (La-WT) versus L. amazonensis arginase knockout (La-arg-) promastigotes and axenic amastigotes. A total of 8253 transcripts were identified in La-WT and La-arg- promastigotes and axenic amastigotes, about 60% of them codifying hypothetical proteins and 443 novel transcripts, which did not match any previously annotated genes. Our RNA-seq data revealed that 85% of genes were constitutively expressed. The comparison of transcriptome and metabolome data showed lower levels of arginase and higher levels of glutamate-5-kinase in La-WT axenic amastigotes compared to promastigotes. The absence of arginase activity in promastigotes increased the levels of pyrroline 5-carboxylate reductase, but decreased the levels of arginosuccinate synthase, pyrroline 5-carboxylate dehydrogenase, acetylornithine deacetylase and spermidine synthase transcripts levels. These observations can explain previous metabolomic data pointing to the increase of L-arginine, citrulline and L-glutamate and reduction of aspartate, proline, ornithine and putrescine. Altogether, these results indicate that arginase activity is important in Leishmania gene expression modulation during differentiation and adaptation to environmental changes. Here, we confirmed this hypothesis with the identification of differential gene expression of the enzymes involved in biosynthesis of amino acids, arginine and proline metabolism and arginine biosynthesis. All data provided information about the transcriptomic profiling and the expression levels of La-WT and La-arg- promastigotes and axenic amastigotes. These findings revealed the importance of arginase in parasite survival and differentiation, and indicated the existence of a coordinated response in the absence of arginase activity related to arginine and polyamine pathways.


Giblets and abdominal fat of pomegranate seed oil fed chickens as a source of bioactive fatty acids.
The aim of present study was to determine whether chickens' (broilers Ross 308, n = 180, sex ratio 1:1) diet modification with different doses of grape or pomegranate seed oil will favourable change fatty acids and cholesterol content in selected giblets (liver and heart) or wastes (adipose tissue). It was also verify whether generated changes would make the giblets and wastes more valuable as dietary components or by-products for food industry. From 22 to 42 day of life, five diets were administered to chickens. Control grower diet (CON) contained 5% of soy oil, whereas in the experimental grower diets part of soy oil (1.5% or 2%) was replaced with specific amount of grape or pomegranate seed oil (GRAP 1.5; GRAP 2.0; POM 1.5; POM 2.0 respectively). Fatty acids and cholesterol content were determined with gas chromatography with flame-ionization detection. Pomegranate seed oil improved fatty acids profile more favourably than grape seed oil, which makes it a valuable additive in chickens' feeding. Abdominal fat of pomegranate seed oil supplemented chickens appeared to be the richest sources of rumenic acid and n3 polyunsaturated fatty acids, which allows to suggest its use in manufacturing of meat products to obtain foodstuffs rich in those essential nutrients. In principal component analysis (PCA), two principal components: PC1 and PC2, which were enough to explain 29.91% of variance of initial variables, allowed to a good separation of chickens fed with both doses of pomegranate seed oil from animals from control and grape seed oil fed groups. Because poultry addresses all nutritional, institutional and consumer requirements, enrichment of giblets in rumenic acid by pomegranate seed oil incorporation into chickens' diet may provide a valuable dietary source of bioactive fatty acids for consumers, especially of low-income countries.


Sleep quality and paranoia: The role of alexithymia, negative emotions and perceptual anomalies.
Recent evidence suggests that sleep problems are associated with psychotic like experiences including paranoia. However, the mechanisms underpinning this association are not well understood and thus studies modelling hypothesised mediating factors are required. Alexithymia, the inability to recognise and describe emotions within the self may be an important candidate. In two separate studies we sought to investigate factors mediating the relationship between sleep quality and paranoia using a cross-sectional design. Healthy volunteers without a mental health diagnosis were recruited (study 1, N = 401, study 2, N = 402). Participants completed a series of measures assessing paranoia, negative emotions, alexithymia and perceptual anomalies in an online survey. In study 1, regression and mediation analyses showed that the relationship between sleep quality and paranoia was partially mediated by alexithymia, perceptual anomalies and negative affect. In contrast, study 2 found that the relationship between sleep quality and paranoia was fully mediated by negative affect, alexithymia and perceptual anomalies. The link between sleep quality and paranoia is unclear and reasons for discrepant results are discussed. Novel findings in this study include the link between alexithymia and paranoia.


Cause-effect linkages between chemicals and populations of mink (Mustela vison) and otter (Lutra canadensis) in the Great Lakes basin.
Following outbreaks of reproductive failure in commercial ranching operations, laboratory experiments showed that mink are extremely sensitive to organochlorine chemicals, particularly PCBs and dioxins. The purpose of this paper is to test the hypothesis that, since wild mink are exposed to these compounds through consumption of Great Lakes fish, they might exhibit reproductive dysfunction and population declines. The otter, another piscivorous animal, should show the same effects. The available information is reviewed according to five epidemiological criteria. Harvest data are presented as a surrogate for the population status of mink and otters in certain locations around the Great Lakes. Data from Ohio show that the mink harvest between 1982 and 1987 from contaminated counties bordering Lake Erie was consistently lower (380 animals per year) than those from counties removed from Lake Erie (850 animals per year), suggesting an effect of chemicals on the status of mink populations. Preliminary studies from Ontario also suggest that mink harvest is lower in potentially "high PCB exposure areas" compared with lower exposure areas. Evidence is also presented on the harvest data for otters taken from four New York State counties adjacent to Lake Ontario and the St. Lawrence River. The harvest data from these four counties show that between 1960 and early 1970 otter harvest remained stable but has since increased. Increased harvest is consistent with improved water quality in Lake Ontario during the past 15 yr. Data relating to strength of association between chemicals and populations of mink and otter are weak and need to be further analyzed. The specificity of the effects of the chemicals on mink reproduction and mortality is well established from toxicological experiments, but there is poor resolution of the information on effects using field data. The strongest case for a causal relationship comes from consideration of the coherence criterion. In conclusion, before a causal link can be drawn between the status of mink and otter populations and exposure to organochlorine chemicals from the Great Lakes, a large amount of research and data analysis needs to be undertaken.


Improved glycosylation of a foreign protein by Tn-5B1-4 cells engineered to express mammalian glycosyltransferases.
The major advantages of using the baculovirus-insect cell system for recombinant protein production are its ability to produce large amounts of recombinant proteins and its ability to provide eucaryotic modifications, such as glycosylation. However, the glycans linked to recombinant glycoproteins produced by this system typically differ from those found on native mammalian products. This is an important problem because glycans on mammalian glycoproteins can influence their functions in many different ways. The inability of baculovirus-infected insect cells to produce glycans identical to those found on native mammalian glycoproteins is due, in part, to the absence of functional levels of certain glycosyltransferases in insect cells. Thus, the purpose of this study was to engineer these activities into Tn-5B1-4, an established insect cell line that is widely used as a host for baculovirus-mediated protein production. Expression plasmids were constructed in which cDNAs encoding mammalian beta1,4-galactosyltransferase and alpha2,6-sialyltransferase were placed under the transcriptional control of a baculovirus immediate early promoter. These plasmids were then used to isolate two different transgenic Tn-5B1-4 derivatives and the biological and biochemical properties of these cell lines were examined. The results show that both of the engineered insect cell lines have improved glycoprotein-processing capabilities, relative to the parental cell line.


A Multi-Technique Investigation of the Complex Formation Equilibria between Bis-Deferiprone Derivatives and Oxidovanadium (IV).
The increasing biomedical interest in high-stability oxidovanadium(IV) complexes with hydroxypyridinone ligands leads us to investigate the complex formation equilibria of VIVO2+ ion with a tetradentate ligand, named KC21, which contains two 3-hydroxy-1,2-dimethylpyridin-4(1H)-one (deferiprone) moieties, and with the simple bidentate ligand that constitutes the basic unit of KC21, for comparison, named L5. These equilibrium studies were conducted with joined potentiometric-spectrophotometric titrations, and the results were substantiated with EPR measurements at variable pH values. This multi-technique study gave evidence of the formation of an extremely stable 1:1 complex between KC21 and oxidovanadium(IV) at a physiological pH, which could find promising pharmacological applications.


Kinetic properties and Na+ dependence of rheogenic Na(+)-HCO3- co-transport in frog retinal pigment epithelium.
1. Na(+)-HCO3- co-transport across the retinal membrane of the frog retinal pigment epithelium was studied by means of double-barrelled pH-selective microelectrodes. Transient changes in the intracellular pH were monitored in response to abrupt changes in the Na+ concentration on the retinal side of the epithelium. 2. The experiments were performed as follows. The Na(+)-HCO3- co-transport was inhibited by perfusing the retinal side of the epithelium with a Na(+)-free solution. The co-transport was then stimulated by changing the perfusate from the Na(+)-free solution to a solution which contained from 5 to 110 mM-Na+. The resulting inward Na(+)-HCO3- co-transport produced an intracellular alkalinization, the initial rate of which was used to calculate the initial rate of Na(+)-HCO3- co-transport, JHCO3-. 3. The Na+ dependence of the Na(+)-HCO3- co-transport was studied at two different values of extracellular pH (7.40 and 7.10), at constant extracellular HCO3- concentration (27.5 mM) and at two different extracellular HCO3- concentrations (27.5 mM and 55 mM) at constant extracellular pH (7.40). In these experiments, the calculated values of JHCO3- followed single Michaelis-Menten kinetics with respect to the extracellular Na+ concentration. 4. The data are consistent with a model in which the co-transporter has a single binding site for the Na+ ion with an apparent affinity constant (apparent Km) of 37 mM. The apparent affinity constant for Na+ was independent of the extracellular concentration of CO3(2-) in the range of 16-65 microM, and of the extracellular HCO3- concentration in the range 27.5-55 mM. 5. The NaCO3- ion-pair hypothesis, in which sodium binds to the co-transporter and is translocated across the cell membrane as the NaCO3- ion pair, was analysed. For stoichiometries 1:2 and 1:3 of the Na(+)-HCO3- co-transport, the NaCO3- ion-pair hypothesis was found incompatible with the data. 6. The intracellular buffer capacity as measured by the CO2 method was 15 mM.


Predicting 30-Day Hospital Readmission Risk in a National Cohort of Patients with Cirrhosis.
Early hospital readmission for patients with cirrhosis continues to challenge the healthcare system. Risk stratification may help tailor resources, but existing models were designed using small, single-institution cohorts or had modest performance. We leveraged a large clinical database from the Department of Veterans Affairs (VA) to design a readmission risk model for patients hospitalized with cirrhosis. Additionally, we analyzed potentially modifiable or unexplored readmission risk factors. A national VA retrospective cohort of patients with a history of cirrhosis hospitalized for any reason from January 1, 2006, to November 30, 2013, was developed from 123 centers. Using 174 candidate variables within demographics, laboratory results, vital signs, medications, diagnoses and procedures, and healthcare utilization, we built a 47-variable penalized logistic regression model with the outcome of all-cause 30-day readmission. We excluded patients who left against medical advice, transferred to a non-VA facility, or if the hospital length of stay was greater than 30 days. We evaluated calibration and discrimination across variable volume and compared the performance to recalibrated preexisting risk models for readmission. We analyzed 67,749 patients and 179,298 index hospitalizations. The 30-day readmission rate was 23%. Ascites was the most common cirrhosis-related cause of index hospitalization and readmission. The AUC of the model was 0.670 compared to existing models (0.649, 0.566, 0.577). The Brier score of 0.165 showed good calibration. Our model achieved better discrimination and calibration compared to existing models, even after local recalibration. Assessment of calibration by variable parsimony revealed performance improvements for increasing variable inclusion well beyond those detectable for discrimination.


Postmastectomy electron-beam chest-wall irradiation in women with breast cancer.
This retrospective study evaluates the results of postmastectomy electron-beam chest-wall irradiation in patients with breast cancer. From 1980 to 1994, 144 women with localized breast cancer received postmastectomy radiotherapy. The chest wall was irradiated by electron beam, 6 to 12 MeV energy, depending on wall thickness, 2.0 Gy daily, 5 times/week for total dose of 50 Gy. Forty-one patients received 16-Gy boosts to the mastectomy scar. In addition, the supraclavicular and axilla areas were irradiated by anterior field with 6-MV photon beam. Median follow-up was 84 months. Fifteen patients (10%) had local-regional recurrence (LRR) and 57 patients (40%) had systemic relapse (SR). Median time from mastectomy to LRR was 20 months and median time to SR was 33 months. Axillary lymph nodes status influenced both LRR and SR. LRR rate was 0% in N0 and 12% in N1 disease; SR rate was 14% in N0 and 45% in N1 disease. Disease-free and overall survival was 58% and 67% in 10 years and 50% and 55% in 20 years, respectively. No cardiac toxicity was related to left chest-wall irradiation. Postmastectomy electron-beam chest-wall irradiation is as effective as photon-beam irradiation in breast cancer.


MiRNA-301b-3p induces proliferation and inhibits apoptosis in AML cells by targeting FOXF2 and regulating Wnt/β-catenin axis.
MiRNA-301b-3p functions as an oncomiRNA or tumor suppressor, and has been reported in various cancer types, including pancreatic, colorectal, oral, hepatocellular and lung cancers. Although the expression of miRNA-301b-3p is upregulated in acute myeloid leukemia (AML), its biological function and precise mechanisms remain unclarified. This study explores the roles of miRNA-301b-3p in AML, with the aim of ascertaining its regulatory action on Wnt/β-catenin axis by targeting Forkhead box F2 (FOXF2). The expression levels of miRNA-301b-3p and FOXF2 were measured by quantitative real-time PCR. The effects of miRNA-301b-3p knockdown and overexpression on cell proliferation were evaluated by CCK8 and cell counting assays, while cell apoptosis was analyzed by flow cytometry. The expression levels of apoptosis-related proteins, including FOXF2, and other targets in Wnt/β-catenin axis were determined by immunoblotting. Possible interaction between miRNA-301-3p and FOXF2 in AML cells was examined by luciferase reporter assays. MiRNA-301b-3p was dramatically upregulated in AML cells, and showed a negative correlation with FOXF2 expression. Downregulation of miRNA-301b-3p suppressed proliferation and promoted apoptosis in AML cells. MiRNA-301b targeted FOXF2 to regulate Wnt/β-catenin axis. In the rescue experiments, FOXF2 overexpression partly reversed the effect of miRNA-301b-3p mimic in AML cells. The current findings demonstrate that miRNA-301b-3p targets FOXF2 to induce proliferation and inhibit apoptosis in AML cells via regulation of Wnt/β-catenin axis.


Withdrawn: Development of a robotic system for prostate interventional therapy.
Withdrawal: Khalil, I. and Omisore, OM. (2021), Development of a robotic system for prostate interventional therapy. Int J Med Robot. Accepted Author Manuscript e2236. doi: http://10.1002/rcs.2236 The above article from The International Journal of Medical Robotics and Computer Assisted Surgery, published online on 29-Jan-2021 on Wiley Online Library (http://wileyonlinelibrary.com) has been withdrawn to allow the authors to register a patent for prostate interventional therapy, by agreement between the Editor-in-Chief and John Wiley & Sons Ltd.


A Nozaki-Hiyama-Kishi Ni(II)/Cr(II) coupling approach to the phomactins.
[reaction: see text] Through a unified synthetic strategy, appropriately functionalized bicyclic starting materials can be elaborated via Ni(II)/Cr(II) macrocylization to [9.3.1] bicycles. Elaboration of these core structures allows access to phomactin C/D analogues and establishes the first synthetic approach to phomactin A affording an intact octahydrochromene/macrocyclic ring system.


[Research progress on circular RNA in oral squamous cell carcinoma].
Circular RNA, a non-coding RNA that forms a covalently closed continuous loop, exists widely in eukaryotic cells. The biogenesis and biological function of this type of RNA indicate that it can play a crucial role in diseases such as tumors, neural system diseases, and cardiovascular diseases; moreover, this RNA may have great potential use as a biomarker in these diseases. Oral squamous cell carcinoma (OSCC) is a common malignancy in oral surgery that is difficult to cure, metastasizes easily, and has poor prognosis. In this review, we summarize the loop-forming mechanisms and functions of circular RNA and describe the progress of current research in the development of oral cancer.


Growth characteristics, grain filling, and assimilate transport in a shrunken endosperm mutant of barley.
The reported inheritance pattern of the seg1 shrunken endosperm mutant of barley (Hordeum vulgare L. cv Betzes) suggests that some defective process in the maternal plant tissues, and not in the endosperm, prevents normal grain filling in the mutant. To identify the physiological mechanism of the mutation, we compared growth, carbon exchange, and assimilate transport of Betzes and seg1 plants. Betzes and seg1 plants did not differ in mean relative growth rate, mean net assimilation rate, or carbon exchange rate. The rate and duration of grain growth of seg1 was lower than Betzes on intact plants and on detached, cultured spikes. Increasing the supply of sucrose in culture media up to 300 mm sucrose did not eliminate differences between normal and mutant grain growth. Translocation of (14)C-labeled assimilates into seg1 grains ceased by 21 days after anthesis, and assimilates were diverted to lower plant parts. In contrast, assimilates were still entering Betzes grains at 29 days after anthesis. Evidence suggests that some maternal spike or grain tissue is affected by the mutation after the onset of grain filling. Identification of the specific seg1 defect may provide information about the cessation of normal grain filling.


Determination of elastic modulus of the components at dentin-resin interface using the ultrasonic device.
The purpose of this study was to determine the elastic moduli of the components at resin-dentin interface with the use of an ultrasonic device. Dentin plates were obtained from freshly extracted bovine incisors with a shape in rectangular form. Resin composites and bonding agents were polymerized and trimmed in the same shape as the dentin specimens. The ultrasonic equipment employed in this study was comprised of a Pulser-Receiver, transducers, and an oscilloscope. Each elastic modulus was determined by measuring the longitudinal and shear wave sound velocities. The mean elastic modulus of mineralized dentin was 17.4 GPa, while that of demineralized dentin was 1.4 GPa. When the demineralized dentin was immersed in bonding agents, the elastic modulus changed to 3.7-4.7 GPa, and these values were significantly higher than those of demineralized dentin. A gradient in elastic modulus was detected as the analysis shifted from the dentin side to the resin composite.


Cobalt(III) complexes of [3(5)] adamanzane, 1,5,9,13-tetraazabicyclo[7.7.3]nonadecane. Report of an inert, chelate hydrogen carbonate ion.
Three cobalt(III) complexes of the macrocyclic tetraamine [3(5)]adamanzane (1,5,9,13-tetraazabicyclo[7.7.3]nonadecane) were isolated as salts. The X-ray crystal structures were solved for the compounds [Co([3(5)]adz)(CO(3))]AsF(6) (1b), [Co([3(5)]adz)(HCO(3))]ZnBr(4).H(2)O (2a), and [Co([3(5)]adz)(SO(4))]AsF(6).H(2)O (3a). The coordination geometry around the cobalt(III) ion is a distorted octahedron with the inorganic ligands at cis-positions. Complex 2 is the second example of a cobalt(III) complex for which the X-ray structure shows a chelate binding mode of the hydrogen carbonate entity. The pK(a) value of the [Co([3(5)]adz)(HCO(3))](2+) ion (2) was determined spectrophotometrically to be 0.27 (25 degrees C, I = 5.0 M). The protonation appears to occur at the noncoordinated carbonyl oxygen atom of the carbonate group, with hydrogen bonding to the crystal water molecule. Evidence is presented for this oxygen atom as the site of protonation in solution as well. In 5.0 M CF(3)SO(3)H a slow reaction of the carbonato complex, quantitatively yielding the [Co([3(5)]adz)(H(2)O)(2)](3+) ion, was observed. k(obs) = 7.9(1) x 10(-)(6) s(-)(1) at 25 degrees C.


Bioimmunotherapeutic targets on angiogenetic blood vessels in solid malignangies.
Physiological angiogenesis is a tightly regulated process that occurs mainly during reproduction, development and wound healing. Although angiogenesis is a continuous process, different consecutive steps can be identified, including: i) release of pro-angiogenetic factors; ii) release of proteolytic enzymes; iii) endothelial cell migration, morphogenesis and proliferation. Angiogenesis is also a hallmark of malignant diseases, and an inverse correlation between tumor vascularity and survival was demonstrated. Thus, strategies aimed at interfering with tumor blood supply by targeting tumor vasculature, presently represent promising new approaches for the treatment of solid malignancies. In fact, at least 30 angiogenetic inhibitors, utilized alone or in combination with other therapeutic agents, are currently being tested in clinical trials in humans. In this paper, we will review current knowledges on selected molecules expressed by endothelial cells and involved in distinct steps of the angiogenetic process, that represent potential targets for bioimmunotherapeutic approaches in human malignancies.


Cancer Vaccine Development Program collaborations.
Immunotherapy for cancer, which uses the body's immune system to fight the disease, is an increasingly active area of research. Successful therapies such as trastuzamab (Herceptin) for breast cancer and cytokine therapy for renal cell carcinoma and melanoma have validated the field as a viable area of investigation. However, the goal of developing an effective cancer vaccine has not yet been achieved. The military's Cancer Vaccine Development Program (CVDP) is collaborating with other military programs, along with civilian institutions, to advance scientific research surrounding cancer vaccines.


Fluorescent and colorimetric probes for mercury(II): tunable structures of electron donor and π-conjugated bridge.
A new series of intramolecular-charge-transfer (ICT) molecules (compounds 1, 2, and 3) were synthesized by attaching various electron-donating thiophenes groups to a triphenylamine backbone with an aldehyde group as the electron acceptor. Based on the protection reaction between ethanethiol and aldehyde, the corresponding dithioacetals (compounds S1, S2, and S3) were prepared to serve as novel colorimetric and fluorescent chemosensors for Hg(2+) ions. Also, compound S1 was further utilized to construct the chemical-reaction-based conjugated polymer probe (PS1) towards Hg(2+) ions. In the presence of as little as 10 nM Hg(2+), compound PS1 displayed an apparent change in the fluorescent intensity. The sensing processes were revealed to be mediated by ICT, as confirmed by time-dependent DFT calculations. Furthermore, compound S1 was successfully applied to microscopic imaging for the detection of Hg(2+) in HeLa cells with ratiometric fluorescent methods.


[Subarachnoid hemorrhage following commencement of danazol treatment in a patient well controlled on warfarin anticoagulation].
A 45-year-old woman with prosthetic valves replacement, was admitted with severe headache and vomiting one month after starting danazol treatment at 300 mg per day. She was receiving long-term anticoagulation with warfarin and dipyridamole, taking 3.5 mg and 300 mg per day respectively. The patient's thrombotest value was less than 6% at the time of admission. Cranial CT revealed subarachnoid hemorrhage. Warfarin and danazol treatment ware discontinued with replenishment of vitamin K. Recovery was uneventful. Danazol is 2, 3 isoxazol derivative of 17-alpha-ethinyl testosterone. As such, it shares the property of C 17 alkylated steroids in potentiating the action of coumarin. It is suggested that danazol affects the turnover of vitamin-K-dependent clotting factors, an impairment of synthesis being a likely mechanism. The possible hazard of the potentiating effect of danazol on warfarin should be widely appreciated.


[Relation between insulin resistance and amount of visceral adipose tissue].
Association of obesity with metabolic and cardiovascular complications depends on the adipose tissue distribution. The role of intraabdominal, i.e. visceral, adipose tissue in pathogenesis of insulin resistance is still not elucidated. The aim of this study was to investigate the relation between insulin resistance and contribution of visceral and subcutaneous fat in a group of women with a wide range of body weight. 62 women (age 21-66 years) among which 32 were non-obese and 30 obese (BMI > 30 kg/m2) were examined. The amount of visceral and subcutaneous fat was evaluated using computerized tomography, total body fat evaluated using bioimpedance, and the degree of insulin resistance was evaluated using glucose disposal (M) during euglycemic hyperinsulinemic clamp. Obese women had lower insulin sensitivity than non-obese (5.88 +/- 2.17 vs 3.32 +/- 1.44 mg/min/kg, p <0.001) and higher absolute amount of visceral fat. However, the relative amount of visceral fat (related to total body fat or subcutaneous fat) was not different between the two groups. In the entire study group, the magnitude of insulin sensitivity did correlate with absolute amount of total and visceral fat, but no correlation with relative amount of visceral fat was found. The results suggest that the absolute amount of fat, either total or visceral, is a stronger predictor of the degree of insulin resistance than the relative contribution of visceral fat.


Is it best to add native shrubs to a coastal sage scrub restoration project as seeds or as seedlings?
Ecological restoration frequently involves the addition of native plants, but the effectiveness (in terms of plant growth, plant survival, and cost) of using seeds versus container plants has not been studied in many plant communities. It is also not known if plant success would vary by species or based on functional traits. To answer these questions, we added several shrub species to a coastal sage scrub restoration site as seeds or as seedlings in a randomized block design. We measured percent cover, density, species richness, size, survival, and costs. Over the two years of the study, shrubs added to the site as seeds grew more and continued to have greater density than plants added from containers. Seeded plots also had greater native species richness than planted plots. However, shrubs from containers had higher survival rates, and percent cover was comparable between the planted and seeded treatments. Responses varied by species depending on functional traits, with deep-rooted evergreen species establishing better from container plants. Our cost analysis showed that it is more expensive to use container plants than seed, with most of the costs attributed to labor and supplies needed to grow plants. Our measurements of shrub density, survival, species richness, and growth in two years in our experimental plots lead us to conclude that coastal sage scrub restoration with seeds is optimal for increasing density and species richness with limited funds, yet the addition of some species from container plants may be necessary if key species are desired as part of the project objectives.