Neem gum based pH responsive hydrogel matrix: A new pharmaceutical excipient for the sustained release of anticancer drug.
The present research work was aimed to synthesize neem gum-based site-specific drug delivery device for anticancer drug methotrexate at different pH condition. The hydrogel-based drug delivery device was synthesized by optimizing reaction parameters using a factorial design approach response surface method. This model comprised of various sets of reactions with varying concentrations of solvent, crosslinker, initiator and monomer under microwave radiation. Characterization of the candidate hydrogel was done using UV-visible spectrophotometer, FTIR, SEM, Raman, and XRD techniques. The release profile of the hydrogels network was studied through a methotrexate under different pH conditions. The drug encapsulation capacity was found to be around 93% and 90% in pH 7.4 and 6.8. Drug release through the synthesized hydrogel matrix was found to show non-Fickian behaviour at each medium. The hydrogel network showed less release in pH 6.8 than pH 7.4, suggesting that hydrogels may be suitable drug carriers for release of anticancer drug delivery system. Hemolysis testing was also done to check the compatibility of the synthesized drug delivery device with the four different blood samples. Hemolysis was found to be less than 1% in the case of all blood groups, which indicates that the synthesized candidate polymers are biocompatible with all blood groups.


The effect of artificial feeding on cholestasis, gallbladder sludge and lithiasis in infants: correlation with plasma cholecystokinin levels.
The occurrence of hepatic cholestatis (judged by fasting serum bile acid levels), gallbladder sludge formation and lithiasis (ultrasonography) and their correlation with plasma cholecystokinin (CCK) levels was studied in a group of children on continuous total parenteral nutrition (TPN) (n = 95), and later in 40 of these children on cyclic TPN (cTPN). After resumption of oral feeding, 75 were studied on partial oral feeding (2-4 meals) and 40 on constant rate enteral nutrition (CREN) then 45 on total oral feeding (4-6 meals). Gallbladder sludge occurred in 23% of the children on TPN for 1 month and 32% of those on cTPN for 3 months. On CREN, the sludge rate was unchanged, but dropped significantly (17%) on partial oral feeding, and disappeared in children on total oral feeding. Serum bile acids were abnormal in 80% of children on TPN or cTPN and diminished significantly on total oral feeding only. Plasma CCK levels on TPN, cTPN and CREN were identical to fasting levels of children on total oral feeding. Plasma CCK levels increased significantly 1 h post-prandially during both partial (p < 0.02) and total oral feeding (p < 0.001). There was a significant negative correlation between the gallbladder sludge rate and CCK levels for all methods of feeding. This study demonstrates the frequent occurrence of hepatic cholestasis in infants, and the much lower frequency of gallbladder sludge in children compared to adults on TPN. Plasma CCK levels obtained during the different methods of feeding could explain the reduction and eventual disappearance of sludge following stimulation of CCK secretion by discontinuous feeding.


Wealth redistribution promotes happiness.
How much happiness could be gained if the world's wealth were distributed more equally? Despite decades of research investigating the relationship between money and happiness, no experimental work has quantified this effect for people across the global economic spectrum. We estimated the total gain in happiness generated when a pair of high-net-worth donors redistributed US$2 million of their wealth in $10,000 cash transfers to 200 people. Our preregistered analyses offer causal evidence that cash transfers substantially increase happiness among economically diverse individuals around the world. Recipients in lower-income countries exhibited happiness gains three times larger than those in higher-income countries. Still, the cash provided detectable benefits for people with household incomes up to $123,000.


Regulation of ASAP1 by phospholipids is dependent on the interface between the PH and Arf GAP domains.
ASAP1 is an Arf GAP with a PH domain immediately N-terminal to the catalytic Arf GAP domain. PH domains are thought to regulate enzymes by binding to specific phosphoinositide lipids in membranes, thereby recruiting the enzyme to a site of action. Here, we have examined the functional relationship between the PH and Arf GAP domains. We found that GAP activity requires the cognate PH domain of ASAP1, leading us to hypothesize that the Arf GAP and PH domains directly interact to form the substrate binding site. This hypothesis was supported by the combined results of protection and hydrodynamic studies. We then examined the role of the PH domain in the regulation of Arf GAP activity. The results of saturation kinetics, limited proteolysis, FRET and fluorescence spectrometry support a model in which regulation of the GAP activity of ASAP1 involves a conformational change coincident with recruitment to a membrane surface, and a second conformational change following the specific binding of phosphatidylinositol 4,5-bisphosphate.


Research agenda for preterm birth: recommendations from the March of Dimes.
Preterm birth (PTB) is a common, serious, and costly health problem affecting nearly 1 in 8 births in the United States. Burdens from PTB are especially severe for the very preterm infant (<32 weeks' gestation), comprising 2% of all US births. Successful prevention needs to include newly focused and adequately funded research, incorporating new technologies and recognition that genetic, environmental, social, and behavioral factors interact in complex pathogeneses and multiple pathways leading to PTB. The March of Dimes Scientific Advisory Committee created this prioritized research agenda, which is aimed at garnering serious attention and expanding resources to make major inroads into the prevention of PTB, targeting six major, overlapping categories: epidemiology, genetics, disparities, inflammation, biologic stress, and clinical trials. Analogous to other common, complex disorders, progress in prevention will require incorporating multipronged risk reduction strategies that are based on sound scientific discovery, as well as on effective translation into clinical care.


Structure of the M2 protein gene of sonchus yellow net virus.
The nucleotide sequence of the gene immediately following the nucleocapsid protein gene of sonchus yellow net virus (SYNV), a plant rhabdovirus, is presented. Serological reactions of SYNV proteins with antibodies elicited by a fusion protein constructed from the sequenced gene indicate that this gene encodes an SYNV structural protein designated M2. The 5' end of the M2 protein mRNA appears to correspond to position 1700 relative to the 3' end of SYNV RNA, because an extension product that maps to this position was synthesized by reverse transcription of polyadenylated [poly(A)+] RNA from infected tobacco that had been primed with an SYNV-specific oligodeoxyribonucleotide. The 3' end of the gene encoding the M2 protein is defined by a recombinant DNA plasmid derived from poly(A)+ RNA from SYNV-infected plants. This plasmid contains an insert with a 3'-terminal region corresponding to a uridylate-rich sequence present at positions 2832 to 2836 on SYNV genomic (g) RNA. These data thus suggest that the M2 protein mRNA is 1132 nucleotides (NT) long, excluding the poly(A) tail, and consists of a 50-NT untranslated 5' region, a 1035-NT open reading frame (ORF), and a 47-NT untranslated 3'region. The ORF is capable of encoding a 345-amino acid protein with a calculated molecular weight of 38,332. A small region of the M2 protein appears to have some similarity to the phosphoproteins of other rhabdoviruses. An identical 14-NT region occurs at the two sequenced gene junctions on SYNV gRNA and shares homology with regions separating the genes of some animal rhabdoviruses.


Activation of phospholipase D in CHO cells transfected with the human epidermal growth factor (EGF) receptor: differential effects of protein kinase C activation and EGF.
Multiple intracellular signal transduction pathways, including phospholipases A2 and D, can be activated by epidermal growth factor (EGF) in both a protein kinase C (PKC)-dependent and -independent manner. We investigated the activation of phospholipase D (PLD) by a PKC activator, phorbol myristate acetate (PMA) and by EGF in CHO cells transfected with the full-length EGF receptor. In cells labelled with arachidonic acid or linoleic acid, PMA activated a PLD, determined by formation of the transphosphatidylation product phosphatidylethanol in the presence of ethanol. A basal PLD activity was seen in linoleic acid-labelled cells but not in cells labelled with arachidonic acid. This basal activity was augmented by the protein phosphotyrosine phosphatase inhibitor vanadate and reduced by tyrosine kinase inhibition and was contributed to by PKC, as activity could not be elicited following prolonged exposure to phorbol ester, known to down-regulate some PKC isoforms. By contrast, EGF failed to stimulate formation of phosphatidylethanol in cells labelled with either fatty acid species. It is proposed that in the basal condition PKC-dependent PLD activation and protein tyrosine kinase phosphorylation are linked (possibly by a phospholipase C (PLC)-mediated formation of diacylglycerol); EGF which activated a phospholipase A2 (PLA2) but which failed to elicit PLC activation in these cells is without further effect on PLD.


Iron deficiency caused by 7 weeks of intensive physical exercise.
The present study was designed to evaluate the effect of an intensive physical training program involving both isometric and isotonic activities on the body iron status of 8 females and 11 males (age 20 +/- 1 year). The training was carried out over a 7 week period and included 8 h of varying physical activities each day. Venous blood samples were obtained from the subjects prior to the beginning of the training, on day 2 and in weeks 2, 4, 6 and 7 of the program. Blood samples were analyzed for iron, ferritin and hemoglobin (Hb) concentrations, total iron binding capacity (TIBC) and red blood cell count (RBC). Iron levels of males and females decreased 65% after 2 weeks of training (p less than 0.001). At the end of the training program 5 males and 6 females had lower than normal iron values (less than 13.4 mumol.l-1). TIBC increased 25% in women and 18% in men following 2 and 4 weeks of training (p less than 0.001) and remained at this elevated level throughout the training period. Ferritin levels decreased 50% in both sexes after 4 weeks of exercise (p less than 0.05) and remained at this level until the end of the training. Hb and RBC decreased 8-10% in both sexes during the training period. In two of the women anemia occurred after 4 weeks of training. The development of latent iron deficiency in a substantial number of participants after a relatively short period of training is uncommon and may reflect the high intensity of exercise required in this program.(ABSTRACT TRUNCATED AT 250 WORDS)


Pretreatment levels of serum squamous cell carcinoma antigen and urine polyamines in women with squamous cell carcinoma of the cervix.
To investigate whether pretreatment levels of serum squamous cell carcinoma antigen (SCCA) and urine polyamines can predict lymph node metastases in patients with early stage cervical carcinoma. Pretreatment measurement of serum SCCA and urine polyamine levels was carried out for 419 women. Of those women, 104 with stage IB and IIA cancer received radical surgery and had tumor size reassessed postoperatively. The women had increased levels of serum SCCA (>2.0 ng/mL) and elevated urine polyamines (>45 micromol/g of creatinine) with advanced cancer stage (P<0.01). The median SCCA level was significantly higher in women with metastatic disease than that in those without lymph node involvement (3.9 vs. 1.1; P<0.01). Women with nodal involvement also had significantly higher median levels of urine polyamines than those without nodal disease. Pretreatment measurement of SCCA and urine polyamine levels may help in predicting lymph node metastases in women with early stage cervical carcinoma.


Anatomic Placement of the Femoral Tunnels in Double-Bundle Anterior Cruciate Ligament Reconstruction Correlates With Improved Graft Maturation and Clinical Outcomes.
To compare maturation of reconstructed graft on second-look arthroscopy and clinical outcomes between 2 groups: the provisional anatomic (PA) group, with both the anteromedial (AM) and posterolateral (PL) femoral tunnels in their anatomic location, and the nonanatomic (NA) group, with either 1 of the 2 femoral tunnels beyond its anatomic location after double-bundle anterior cruciate ligament reconstruction. We enrolled 154 patients who underwent 3-dimensional computed tomography scanning and second-look arthroscopy after double-bundle anterior cruciate ligament reconstruction. All of the patients were divided into the PA and NA groups according to the femoral tunnel position determined by the quadrant method. Graft maturation was evaluated with 3 subsections, including integrity, tension, and synovial coverage with revascularization, on second-look arthroscopy. We also compared Lachman test, pivot-shift test, KT-2000 (MEDmetric, San Diego, CA), and International Knee Documentation Committee grades at the last follow-up. Of the 154 patients, 88 were classified as the PA group and 66 as the NA group by the quadrant method. A difference existed between groups for the AM tunnel position but not for the PL tunnel position. The PA group showed a higher graft maturation score (P < .001 for all comparisons) and better results according to the International Knee Documentation Committee knee rating, Lachman test, pivot-shift test, and KT-2000 assessment (P < .001 for all comparisons). The PA group with anatomic femoral tunnel placement showed a higher graft maturation score on second-look arthroscopy, along with better clinical outcomes, than the NA group. There was a significant difference in the AM femoral tunnel position but not in the PL tunnel position between the 2 groups. Level III, retrospective comparative study.


Anti-cancer activity of two novel heterocyclic compounds through modulation of VEGFR and miR-122 in mice bearing Ehrlich ascites carcinoma.
Metastasis in breast cancer is a leading cause of mortality among women in many countries. This study investigated the anti-cancer role of benzoimidazoquinazoline and benzimidazotriazin; two novel compounds that were designed, synthesized, structurally elucidated, and biologically evaluated as potent anti-angiogenic agents that act through inhibition of vascular endothelial growth factor receptor-2 (VEGFR2). Breast cancer was induced by inoculation of Ehrlich Ascites Carcinoma (EAC) cells. Seventy swiss albino mice were randomly divided into 7 groups, 10 animals each: (1) normal, (2) control EAC group, (3) cisplatin treated group, (4&5) benzoimidazoquinazoline treated (5 mg/kg and 10 mg/kg), (6&7) benzimidazotriazin treated (5 mg/kg and 10 mg/kg). The expression of miR-122 was assessed in the tumor tissue by quantitative PCR, and the VEGF level was determined in serum by ELISA. VEGFR2 and cluster of differentiation (CD)34 were assessed by immunohistochemistry. Serum ALT, AST, creatinine, and urea were measured. Treatment with benzoimidazoquinazoline and benzimidazotriazin decreased tumor weight and serum levels of VEGF, and down-regulated expression of VEGFR2 and CD34 in the tumor tissue. miR-122 was upregulated, particularly in the benzimidazotriazin (10 mg/kg) group. Relative to cisplatin, the novel compounds were less toxic to kidneys. Benzoimidazoquinazoline and benzimidazotriazin are promising anti-cancer agents that act through inhibition of angiogenesis and thus provide a new strategy for advancement of chemotherapy.


Hepatoprotective activity of two plants belonging to the Apiaceae and the Euphorbiaceae family.
The different extracts of Apium graveolens Linn. (Apiaceae) and Croton oblongifolius Roxb. (Euphorbiaceae) were tested for their hepatoprotective activity against CCl(4) induced hepatotoxicity in albino rats. The degree of protection was measured by using biochemical parameters like serum transaminases (SGOT and SGPT), alkaline phosphatase, total protein and albumin. The methanolic extracts showed the most significant hepatoprotective activity comparable with standard drug silymarin. Other extracts namely petroleum ether and acetone also exhibited a potent activity.


Tourism and glocalization "Local" Tour Guiding.
In tourism studies globalization and localization are often conceived of as a binary opposition. The ethnography of an Indonesian group of tour guides presented here illustrates how the global and the local are intimately intertwined through what has been described as the process of "glocalization". The guides studied are remarkable front-runners of glocalization. They fully participate in global popular culture and use new technologies in their private lives. While guiding, however, they skillfully represent the glocalized life around them as a distinctive "local", adapted to the tastes of different groups of international tourists. It is concluded that tourism offers excellent opportunities to study glocalization, but that more grounded research is needed.


Prevalence and risk factors of excessive daytime sleepiness in major depression: A study with 703 individuals referred for polysomnography.
To date, few studies have investigated the prevalence and risk factors of excessive daytime sleepiness (EDS) in major depression. Thus, the aim of the present study was to examine the prevalence and risk factors of EDS in a large sample of individuals with major depression. Data from 703 individuals with major depression were retrospectively collected from the sleep laboratory research database of Erasme Hospital for analysis. A score of > 10 on the Epworth Sleepiness Scale was used as the cut-off for EDS. Logistic regression analyses were conducted to examine the clinical and demographic risk factors of EDS in major depression. The prevalence of EDS in our sample was 50.8%. Multivariate logistic regression analysis revealed that the following were significant risk factors of EDS in major depression: non-use of short to intermediate half-life benzodiazepine receptor agonists, BMI ≥ 25 kg/m², age < 60 years, C-reactive protein > 7 mg/L, Beck Depression Inventory score ≥ 16, atypical depression, apnea-hypopnea index ≥ 15/h, and use of selective serotonin reuptake inhibitors or serotonin-norepinephrine reuptake inhibitors. To evaluate EDS, we used the Epworth Sleepiness Scale, which only allows for a subjective measure of daytime sleepiness. EDS is a common symptom in individuals with major depression. In this subpopulation, interventions are possible for most risk factors of EDS, which justifies improved management of this symptom to avoid its negative consequences.


The donor health assessment questionnaire: potential for format change and computer-assisted self-interviews to improve donor attention.
The Canadian donor health assessment questionnaire (DHAQ) has developed gradually over many years. The purpose of this study was to determine whether the format and method of administration of the DHAQ influences donor attentiveness. Between May 2004 and September 2005, a total of 1,397 donors participated in the study by completing the DHAQ by one of three methods: the current Canadian Blood Services (CBS) format and method of administration, which is partly self-administered and partly interviewer-administered (Method 1); the DHAQ reformatted to the AABB Uniform Donor Health Questionnaire format and self-administered (Method 2); and an audiovisual computer-assisted self-interview (CASI; Method 3). This was followed by a short, scripted interview assessing recall of 17 specific items queried on the DHAQ. Time to completion of the DHAQ and degree of familiarity with computer use were also assessed. The percentages of donors identifying all 17 items correctly were 9.4, 20.9, and 34.8 percent and the mean percentages recall of items were 53.9, 56.0, and 69.8 percent with DHAQ administration Methods 1, 2, and 3, respectively (p < 0.0001). This difference was largely attributable to the poor recall of items queried as part of a list in Method 1. Mean times to complete the DHAQ were 3.1, 3.8, and 8.1 minutes for Methods 1, 2, and 3, respectively. More than 95 percent of donors had used a personal computer in the past year. The current format of the Canadian DHAQ is not optimal for donor attention to specific questions asked as part of a list. Attention was improved by use of AABB uniform donor history questionnaire format and was best with use of a CASI format.


Experimental studies on the disappearance of subretinal fluid after episcleral buckling procedures without drainage.
We have developed an experimental model to show that an additional mechanism apart from absorption by pigment epithelium is responsible for the absorption of subretinal fluid. The cause of a fast diminution of subretinal fluid after a buckling procedure is the prominence of the buckle as an obstacle to fluid movements in the eye. This results in any fluid being pushed through the retinal hole into the vitreous. When the retinal hole is closed by the buckle, the remaining part of the subretinal fluid is slowly absorbed by the pigment epithelium present. We performed a preliminary qualitative and quantitative analysis of the phenomena occurring using the finite element method of Zienkiewicz.


[Detection and evaluation of drug addict patients seen at a primary care center].
Description of drug-dependent patients who attend a primary care centre (PCC). A prospective study. Barceloneta PCC, located in a waterside area of Barcelona's old quarter. The analysis was carried out on 43 drug-dependent patients who attended the PCC over a two-year period requesting detoxication treatment as well as on patients who attended for treatment of other organic pathologies or to request medication and in whom an addiction problematic was detected. Patients who were already following a course of treatment in a specialised drug dependency centre or who were dependent on alcohol were excluded. Detection and evaluation of drug dependency and of hygienic-dietary problems. A structured interview which looked at the drug history, sociodemographic characteristics and hygienic-dietary habits was employed. It was observed that 90.7% of the drug-dependent patients used heroin and 44% of these had not been previously treated for addiction. 88.4% were male and 51.2% had a criminal record. 72% practised one of the types of behaviour putting them at risk of HIV infection. The majority of patients addicted to drugs who were identified at a PCC are dependent on heroin as their primary drug. 44% of them had received no previous treatment for their addiction.


Microbial complexes levels in conventional and self-ligating brackets.
The aims were to evaluate the levels of bacterial species in saliva and in situ and to assess whether the design of brackets influences the risk of developing periodontal disease. Twenty patients (13.3 mean age) were bonded with self-ligating brackets and a conventional bracket. Saliva was collected before bonding and 30 and 60 days after bonding. One sample of each bracket was removed 30 and 60 days after bonding. The analysis was determined by checkerboard DNA-DNA hybridization. The data was evaluated by the non-parametric test. A significant increase in the levels of bacterial species in the saliva occurred in 15 of the 22 analyzed species. The self-ligating brackets presented the highest incidence percentages for the orange and red complexes 60 days after bonding. In situ analyses showed different patterns according to the bracket design. The levels of Campylobacter rectus showed significant differences (p = 0.011) 60 days after bonding among the three brackets; the highest values were observed in the In-Ovation®R bracket. The bracket design seems to influence the levels of bacterial species involved in periodontal disease. Considering the wide variety of bacterial species, additional studies are needed to aid in the establishment of effective protocols to prevent the development of periodontal disease during orthodontic treatment. A dynamic alteration in the oral microbiota may lead to inflammatory reactions in the supporting soft and hard tissues. The different types of brackets interfere with bacterial adherence. Bracket design should be considered in orthodontic treatment.


Protein arginine deiminase 2 (PAD2) modulates the polarization of THP-1 macrophages to the anti-inflammatory M2 phenotype.
Macrophages are effector cells of the innate immune system that undergo phenotypical changes in response to organ injury and repair. These cells are most often classified as proinflammatory M1 and anti-inflammatory M2 macrophages. Protein arginine deiminase (PAD), which catalyses the irreversible conversion of protein-bound arginine into citrulline, is expressed in macrophages. However, the substrates of PAD and its role in immune cells remain unclear. This study aimed to investigate the role of PAD in THP-1 macrophage polarization to the M1 and M2 phenotypes and identify the citrullinated proteins and modified arginines that are associated with this biological switch using mass spectrometry. Our study showed that PAD2 and, to a lesser extent, PAD1 and PAD4 were predominantly expressed in M1 macrophages. We showed that inhibiting PAD expression with BB-Cl-amidine decreased macrophage polarization to the M1 phenotype (TNF-α, IL-6) and increased macrophage polarization to the M2 phenotype (MRC1, ALOX15). This process was mediated by the downregulation of proteins involved in the NF-κβ pathway. Silencing PAD2 confirmed the activation of M2 macrophages by increasing the antiviral innate immune response and interferon signalling. A total of 192 novel citrullination sites associated with inflammation, cell death and DNA/RNA processing pathways were identified in M1 and M2 macrophages. We showed that inhibiting PAD activity using a pharmacological inhibitor or silencing PAD2 with PAD2 siRNA shifted the activation of macrophages towards the M2 phenotype, which can be crucial for designing novel macrophage-mediated therapeutic strategies. We revealed a major citrullinated proteome and its rearrangement following macrophage polarization, which after further validation could lead to significant clinical benefits for the treatment of inflammation and autoimmune diseases.


A potent drug candidature of Cu(II) pyrazino[2,3-f][1,10]phenanthroline complexes with bioactive ligands: synthesis, crystal structures, biomolecular interactions, radical scavenging and cytotoxicities.
A novel ternary copper(II) complexes, - [Cu(py-phen)(asn)(NO3)(H2O)] (1) and [Cu(py-phen)(trp)(H2O)]NO3 (2)- (py-phen: pyrazino[2,3-f][1,10]phenanthroline, asn: asparagine, trp: tryptophan), have been synthesized and characterized by CHN analysis, ESI-MS, FTIR and single-crystal X-ray diffraction techniques. Interaction of the complexes 1 and 2 with CT-DNA has been investigated by absorption spectral titration, EB and Hoechst 33258 displacement assay. The interaction between the complexes 1 and 2 and BSA was investigated by electronic absorption and fluorescence spectroscopy methods. The experimental outcomes indicate that the fluorescence quenching mechanism between the complexes 1 and 2 and BSA is a static quenching process. The Stern-Volmer constants, binding constants, binding sites and the corresponding thermodynamic parameters (ΔG, ΔH, ΔS) of BSA + complex systems were determined at different temperatures. The binding distance between the complexes 1 and 2 and BSA was calculated according to FRET. The effect of the complexes 1 and 2 on the conformation of BSA was also examined using synchronous, two dimensional (2D) and three dimensional (3D) fluorescence spectroscopy. Radical scavenging activity of the complex was determined in terms of EC50, using the DPPH and H2O2 method. The anticancer activities of the complexes 1 and 2 were investigated using an XTT assay against three cancer cell lines (MCF-7, Caco-2 and A549) and non-tumor cell line (BEAS-2B). Communicated by Ramaswamy H. Sarma.


Sequence similarities among monkey ori-enriched (ors) fragments.
Nucleotide sequences have been determined for eight ors (ori-enriched sequence) fragments isolated from monkey DNA by a method that was designed to enrich for origins of DNA replication [Kaufmann et al., Mol. Cell. Biol. 5 (1985) 721-727]. Evidence has been presented that some or possibly all of these sequences can serve, albeit inefficiently, as oris in vivo [Frappier and Zannis-Hadjopoulos, Proc. Natl. Acad. Sci. USA 84 (1987) 6668-6672]. Two of the fragments were found to contain the long terminal repeat-like elements of the 'O-family' of moderately repetitive sequences that are present in human DNA as a transposon-like element [Paulson et al., Nature 315 (1985) 359-361]. Extensive pair-wise comparisons of the sequences failed to detect any statistically significant common sequences, except for long asymmetrically distributed A + T-rich stretches. Nonetheless, when the ors fragments were examined for the presence of published consensus sequences, seven of eight were found to contain the control sequence described by Dierks et al. [Cell 32 (1983) 695-706], and the same seven of eight were found to contain both the scaffold attachment region T consensus [Gasser and Laemmli, Cell 46 (1986) 521-530] and the minimal Saccharomyces cerevisiae autonomously replicating sequence consensus [e.g., Palzkill and Newlon, Cell 53 (1988) 441-450].


Scaffold design for artificial tissue with bone marrow stem cells.
The aim of this study was to test polymeric materials (collagen, fibrin, polyimide film, and polylactic acid) for single- and multi-layer scaffold formation. In our study, we used rabbit bone marrow stem cells (rBMSCs) and human mesenchymal stem cells (hMSCs) with materials of a different origin for the formation of an artificial scaffold, such as a collagen scaffold, fibrin scaffold produced from clotted rabbit plasma, electrospun poly(lactic acid) (PLA) mats, polyimide film (PI), and the combination of the latter two. Cell imaging was performed 3-14 days after cell cultivation in the scaffolds. Time-lapse imaging was used to determine hMSC mobility on the PI film. Cell incorporation in collagen and clotted fibrin scaffolds was evaluated after 2-week cultivation in vitro. Histological analysis showed that cells penetrated only external layers of the collagen scaffold, while the fibrin clot was populated with rBMSCs through the entire scaffold thickness. As well, cell behavior on the laser micro-structured PI film was analyzed. The mobility of hMSCs on the smooth PI film and the micro-machined surface was 20±2μm/h and 18±4μm/h, respectively. After 3-day cultivation, hMSCs were capable of spreading through the whole 100±10μm-thick layer of the electrospun PLA scaffold and demonstrated that the multilayer scaffold composed of PI and PLA materials ensured a suitable environment for cell growth. The obtained results suggest that electrospinning technology and femtosecond laser micro-structuring could be employed for the development of multi-layer scaffolds. Different biopolymers, such as PLA, fibrin, and collagen, could be used as appropriate environments for cell inhabitation and as an inner layer of the multi-layer scaffold. PI could be suitable as a barrier blocking cell migration from the scaffold. However, additional studies are needed to determine optimal parameters of inner and outer scaffold layers.


Postjuvenile molt in east African and Central European stonechats (Saxicola torquata axillaris, S.t. rubicula) and its modification by photoperiod.
Twenty-eight stonechats of the European race (Saxicola torquata rubicula) from Austria and thirty-one stonechats of the Central African race (S.t. axillaris) from equatorial Kenya were handraised and subsequently investigated with regard to the temporal pattern of their postjuvenile molt. About one half of the birds of each race were held under their own native photoperiod and the other half under the photoperiodic conditions of the other race. The results demonstrated clear differences in the postjuvenile molt between the two races when birds were kept in the photoperiod under which they normally live. The African birds began to molt earlier and molted longer than their European conspecifics. The time course of postjuvenile molt was affected by photoperiod in both races as molt began and ended earlier under the equatorial photoperiod than under the European photoperiodic simulation. The question why the African birds showed strong photoperiodic reactions under these experimental conditions although in their natural environment they experience only minute photoperiodic variations, is critically evaluated: Three possible explanations are discussed: (1) the photoperiodic reaction may represent a relict from a time when ancestors of the tropical populations still lived in more temperate zones; (2) it may be due to the occasional immigration of conspecifics from populations living further north or south; (3) it may result from effects on a (possibly circadian) submechanism of the system controlling annual cycles which is normally affected by other environmental cues but can also be influenced by photoperiodic variations.


White shrimp Litopenaeus vannamei that have received mixtures of heat-killed and formalin-inactivated Vibrio alginolyticus and V. harveyi exhibit recall memory and show increased phagocytosis and resistance to Vibrio infection.
Heat-killed Vibrio alginolyticus (HVa), formalin-inactivated V. alginolyticus (FVa), heat-killed Vibrio harveyi (HVh), formalin-inactivated V. harveyi (FVh), live V. alginolyticus (LVa), and live V. harveyi (LVh) were used in this study. White shrimp Litopenaeus vannamei receiving two mixtures (HVa + FVa) or four mixtures (HVa + FVa + HVh + FVh) served as primary exposure, and shrimp receiving LVa or LVh afterward served as secondary exposure. Shrimp receiving marine saline and then receiving either LVa or LVh served as controls. Phagocytic activity and clearance efficiency were examined in shrimp that received two mixtures after 1-8 weeks and then received LVa. Both the phagocytic activity and clearance efficiency of shrimp receiving two mixtures were significantly higher than in control shrimp after 1-8 weeks. In another experiment, phagocytic activity and clearance efficiency were examined in shrimp that received four mixtures after 1-8 weeks and then received LVa and LVh, respectively. The phagocytic activity of shrimp receiving four mixtures was significantly higher than in control shrimp after 1-8 weeks post exposure to LVa and LVh. The clearance efficiency of shrimp receiving four mixtures was significantly higher than in control shrimp after 1-6 weeks post exposure to LVa, and 1-7 weeks post exposure to LVh. In the other experiment, the survival rate of shrimp that received four mixtures after five weeks were challenged with LVa at 6.4 × 107 colony-forming units (cfu) shrimp-1 and LVh at 4.4 × 106 cfu shrimp-1. Shrimp that received marine saline for five weeks and then challenged with LVa and LVh at a same dose served as challenged controls. The survival rate of shrimp that received four mixtures was significantly higher (90%) than that of control shrimp (67%), and significantly higher (73%) than that of control shrimp (53%) after 3-7 days post challenge with LVa and LVh. It is concluded that the mixtures have feature of adjuvant and antigen, and shrimp receiving mixtures of heat-killed and formalin-inactivated V. alginolyticus and V. harveyi even after 5-8 weeks exhibit memory recall and show increased phagocytosis and resistance to Vibrio infections.


How should I treat stent dislodgement in a STEMI patient resulting in dissection of left main and left circumflex arteries?
A 52-year-old female presented with acute anterior ST-elevation myocardial infarction (STEMI) within one hour of symptom onset to the emergency department. She was referred for urgent primary angioplasty. Physical examination, laboratory investigations, ECG, urgent percutaneous coronary intervention (PCI). Single-vessel coronary artery disease (SVD). Intended to stent culprit lesion. However, stent dislodged in left main coronary artery (LMCA) during attempted PCI to diffuse mid segment of left anterior descending (LAD). Initial attempt failed to retrieve the dislodged stent with snare. Dislodged stent removed with multiple wire technique, complicated by severe dissection in LAD and left circumflex artery back into the LMCA. The stent was trapped at tip of 6 Fr right femoral sheath, unable to be withdrawn. What next?


Injectable LiNc-BuO loaded microspheres as in vivo EPR oxygen sensors after co-implantation with tumor cells.
Electron paramagnetic resonance (EPR) oximetry is a technique which allows accurate and repeatable oxygen measurements. We encapsulated a highly oxygen sensitive particulate EPR spin probe into microparticles to improve its dispersibility and, hence, facilitate the administration. These biocompatible, non-toxic microspheres contained 5-10 % (w/w) spin probe and had an oxygen sensitivity of 0.60 ± 0.01 µT/mmHg. To evaluate the performance of the microparticles as oxygen sensors, they were co-implanted with syngeneic tumor cells in 2 different rat strains. Thus, tissue injury was avoided and the microparticles were distributed all over the tumor tissue. Dynamic changes of the intratumoral oxygen partial pressure during inhalation of 8 %, 21 %, or 100 % oxygen were monitored in vivo by EPR spectroscopy and quantified. Values were verified in vivo by invasive fluorometric measurements using Oxylite probes and ex vivo by pimonidazole adduct accumulation. There were no hints that the tumor physiology or tissue oxygenation had been altered by the microparticles. Hence, these microprobes offer great potential as oxygen sensors in preclinical research, not only for EPR spectroscopy but also for EPR imaging. For instance, the assessment of tissue oxygenation during therapeutic interventions might help understanding pathophysiological processes and lead to an individualized treatment planning or the use of formulations with hypoxia triggered release of active agents.


Variation in immunoreactive trypsinogen concentrations among Michigan newborns and implications for cystic fibrosis newborn screening.
To investigate variation in immunoreactive trypsinogen (IRT) concentrations by race, sex, birth weight, and gestational age and their implications for the use of percentile-based cutoffs for cystic fibrosis (CF) newborn screening (NBS) programs. This cross-sectional population-based study of resident infants screened in Michigan investigates associations between demographic and perinatal variables and IRT concentrations after controlling for covariates. This study also analyzed how 96th and 99.8th IRT concentration percentiles values calculated by Michigan NBS vary by demographic and perinatal factors. Characteristics of infants having high (≥99.8th percentile) IRT concentrations and negative DNA tests are also explored. IRT mean concentrations and percentiles vary significantly by race, birth weight, gestational age, and to a lesser degree by sex. The greatest variation in mean IRT concentrations was observed among racial categories; black infants had an adjusted mean concentration of 36 ng/ml and Asian/Pacific Islander infants had a mean concentration of 25 ng/ml compared to an average concentration of 28 ng/ml in white infants and infants of other races. Variation in IRT concentrations resulted in the over-representation of certain groups referred for secondary testing, particularly referrals for sweat testing based on very high (≥99.8th percentile) concentrations alone, which is no longer recommended in Michigan. Further research may be warranted to evaluate initial IRT cutoffs used for CF NBS.


The preparation and in vivo applications of caged peptides and proteins.
Cellular behavior, such as mitosis and motility, are controlled by both when and where specific intracellular signaling pathways are activated in response to environmental cues. Analogous temporally and spatially controlled events occur throughout the lifetime of an organism (e.g. embryogenesis). Consequently, reagents that can be switched on (or off) at any time or at any place in a cell, a tissue, or a living animal, represent the means by which the biochemical basis of spatially and temporally sensitive biological behavior can be evaluated. This review summarizes recent advances in the design and synthesis of light-activated ('caged') peptides and proteins as well as the application of these caged reagents to unanswered questions in biology.


MicroRNA-19a/b regulates multidrug resistance in human gastric cancer cells by targeting PTEN.
Multidrug resistance (MDR) is the major cause of failure of gastric cancer chemotherapy. Members of the miR-17-92 cluster, including miR-19a/b, are considered oncomiRs and influence multiple aspects of the malignant phenotype of gastric cancer. However, the role of miR-19a/b in MDR in gastric cancer and its underlying mechanism remain unclear. In this study, we found that miR-19a/b were upregulated in MDR cell lines. Our results also showed that miR-19a/b upregulation decreased the sensitivity of gastric cancer cells to anticancer drugs. We further confirmed that miR-19a/b accelerated the ADR efflux of gastric cancer cells by increasing the levels of mdr1 and P-gp and that miR-19a/b suppressed drug-induced apoptosis by regulating Bcl-2 and Bax. Finally, we verified that PTEN, an inhibitor of AKT phosphorylation, is the functional target of miR-19a/b. Overall, these findings demonstrated that miR-19a/b promote MDR in gastric cancer cells by targeting PTEN.


Crystallization and preliminary crystallographic data of chicken gizzard G-actin . DNase I complex and Physarum G-actin . DNase I complex.
Smooth muscle G-actin from chicken gizzard and Physarum plasmodium G-actin both interact with DNase I and form 1 : 1 complexes. These complexes were crystallized by using polyethylene glycol 6000 as a precipitant. Both crystals belong to the same orthorhombic space group P2(1)2(1)2(1). The cell dimensions of chicken gizzard G-actin.DNase I complex are a=42.00 +/- 0.07 A, b=225.3 +/- 0.4 A, and c=77.4 +/- 0.1 A, while those of Physarum G-actin.DNase I complex are a=42 A, b=221 A, and c=77 A.


In vitro biomechanical study of femoral torsion disorders: effect on moment arms of thigh muscles.
Lower limb torsion disorders have been considered as a factor inducing gonarthrosis and the three-dimensional effect of the surgical correction is not well reported yet. This paper reports an in vitro study aiming at quantifying the relationships between experimental femoral torsion disorders and moment arms of thigh muscles. Five unembalmed lower limbs were used and fixed on an experimental jig. Muscles were loaded and 6 Linear Variable Differential Transformers were used to measure tendon excursions. Experimental osteotomies were performed to simulate torsions by steps of 6° up to 18°. Moment arms of the main thigh muscles were estimated by the tendon excursion method during knee flexion. Moment arms of the tensor of fascia latae, the gracilis and the semitendinosus were significantly influenced by experimental conditions while the rectus femoris, the biceps femoris and the semimembranosus did not show modifications. Medial femoral torsion decreased the moment arm of both the gracilis and the semimembranosus. Opposite changes were observed during lateral femoral torsion. The moment arm of the tensor of fascia latae decreased significantly after 30° of knee flexion for 18° of medial femoral torsion. Our results showed that medial and lateral femoral torsion disorders induced alterations of the moment arms of the muscles located medially to the knee joint when applied in aligned lower limbs. These results highlight a potential clinical relevance of the effect of femoral torsion alterations on moment arms of muscles of the thigh which may be related, with knee kinematics modifications, to the development of long-term knee disease.


Sca-1+ cardiac progenitor cell therapy with cells overexpressing integrin-linked kinase improves cardiac function after myocardial infarction.
This study was to investigate the effect of integrin-linked kinase (ILK) on the transplantation efficiency of stem cell antigen-1-positive cardiac progenitor cells (Sca-1 CPCs) in a mouse myocardial infarction (MI) model. Sca-1 CPCs were isolated from C57/BL6 mice heart tissues and genetically modified with adenovirus vector containing green fluorescent protein (GFP)/ILK or GFP. Cell viability, migration, DNA synthesis, proliferation, and apoptosis were assessed in vitro. Immediately after MI, treated animals received 5×10 GFP-CPC or ILK-CPC transplantation into the peri-infarct myocardium. Cardiac function, exercise ability, cardiac morphology, angiogenesis, cardiomyocyte apoptosis, as well as ILK-related protein expression were measured. Acute and long-term cell survival after cell transplantation was assessed. Overexpression of ILK increased the viability, migration, DNA synthesis, proliferation, and survival of Sca-1 CPCs in vitro. Protein expression of phosphorylated Akt and cyclin D1 were up-regulated. In our in vivo experiment, more transplanted cells were found in the peri-infarct myocardium in ILK-CPC group 3 days after cell transplantation, but there was no difference between the two groups 4 weeks later. ILK-CPC group showed reduced infarct size 7 days after cell transplantation. Long-term observation showed improved cardiac function indicated by higher percent fractional shortening and lower left ventricular end systolic diameter/left ventricular end diastolic diameter, better exercise ability, increased angiogenesis, decreased fibrosis and apoptosis, as well as up-regulation of ILK, Cdc42, and Aurora B protein expression in ILK-CPC group 4 weeks after cell transplantation. ILK-overexpressed Sca-1 CPCs showed improved therapeutic efficacy in MI.


The rise of the clinical teaching fellow: a personal view of the postgraduate experience.
Clinical teaching fellowships are becoming increasingly popular, with the numbers of posts ever-expanding. This increase has accelerated as education and training start to recover from the effects of the COVID-19 pandemic. However, the nature of these roles, the entry requirements and the potential benefits are often poorly defined. This article outlines the author's experience of working as a clinical teaching fellow for a year and provides tips on what to look out for when considering these roles, as well as how to get the most out of them.


Mammalian SIRT1 limits replicative life span in response to chronic genotoxic stress.
The Saccharomyces cerevisiae chromatin silencing factor Sir2 suppresses genomic instability and extends replicative life span. In contrast, we find that mouse embryonic fibroblasts (MEFs) deficient for SIRT1, a mammalian Sir2 homolog, have dramatically increased resistance to replicative senescence. Extended replicative life span of SIRT1-deficient MEFs correlates with enhanced proliferative capacity under conditions of chronic, sublethal oxidative stress. In this context, SIRT1-deficient cells fail to normally upregulate either the p19(ARF) senescence regulator or its downstream target p53. However, upon acute DNA damage or oncogene expression, SIRT1-deficient cells show normal p19(ARF) induction and cell cycle arrest. Together, our findings demonstrate an unexpected SIRT1 function in promoting replicative senescence in response to chronic cellular stress and implicate p19(ARF) as a downstream effector in this pathway.


The effects of in vivo inactivation of GABA-transaminase and glutamic acid decarboxylase on levels of GABA in the rat retina.
Gabaculine and gamma-vinyl GABA (GVG) are specific enzyme-activated irreversible inhibitors of GABA-transaminase (GABA-T). gamma-Acetylenic GABA (GAG) irreversibly inhibits both GABA-T and glutamic acid decarboxylase (GAD). Subcutaneous injection of any of those compounds rapidly elevated levels of GABA in the retinae of rats. After injection of 10 mg/kg gabaculine, levels of retinal GABA climbed 5-fold in 4 h, and peaked 16 h after injection at levels approximately 7 times those from water-injected control rats. They remained significantly elevated compared to control levels for at least 6 days after injection. The postgabaculine increase in levels of retinal GABA was linear with time between 0.5 and 4 h after injection. In contrast, retinal GABA levels peaked at less than 3 times control levels within 8 h of injection of 50 mg/kg GAG and returned to baseline levels within 4 days. GAG, upon coadministration with gabaculine, significantly attenuated the postgabaculine rise in levels of GABA in retinae. Neither the rate of rise, nor the maximum levels, of retinal GABA was so great after injection of GAG plus gabaculine, compared to those after injection of gabaculine alone. The degree to which postgabaculine GABA accumulation was inhibited in the retina by 50 mg/kg GAG closely corresponded with the extent to which that dose of GAG inactivated retinal GAD activity. The results of this study extend previous reports from this laboratory that systemically administered gabaculine, GVG and GAG all inactivate target enzymes more potently in retina than in other brain regions.(ABSTRACT TRUNCATED AT 250 WORDS)


Pyruvate kinase from human skeletal muscle.
A simple method is described for the isolation of crystalline pyruvate kinase from human skeletal muscle. The enzyme was purified by ammonium sulfate fractionation, heat treatment and crystallization. Two crystal forms of pyruvate kinase differing in solubility but not in specific activity were found. The homogenous enzyme preparations in triethanolamine buffer, pH 7.6 reveal at 25 degrees a specific activity of 245 U per mg protein, and of 340 U/mg in potassium phosphate buffer (50 mM). The enzyme is activated by inorganic phosphate and fructosediphosphate to the same extent, and inhibited non competetively by ammonium ion. The molecular weight as measured by gel filtration is 220,000 daltons and the enzyme molecule is composed of 4 subunits.


Association of the ACTN3 R577X polymorphism with power athlete status in Russians.
The alpha-actinin-3 (ACTN3) gene encodes a Z-disc structural protein which is found only in fast glycolytic muscle fibers. A common nonsense polymorphism in codon 577 of the ACTN3 gene (R577X) results in alpha-actinin-3 deficiency in XX homozygotes. Previous reports have shown a lower proportion of the ACTN3 XX genotype in power-oriented athletes compared to the general population. In the present study we tested whether XX genotype was under-represented in Russian power-oriented athletes. The study involved 486 Russian power-oriented athletes of regional or national competitive standard. ACTN3 genotype and allele frequencies were compared to 1,197 controls. The frequencies of the ACTN3 XX genotype (6.4 vs. 14.2%; P < 0.0001) and X allele (33.3 vs. 38.7%; P = 0.004) were significantly lower in power-oriented athletes compared to controls. Furthermore, the lowest (3.4%) frequency of the ACTN3 XX genotype was found in a group of highly elite athletes, supporting the hypothesis that the presence of alpha-actinin-3 has a beneficial effect on the function of skeletal muscle in generating forceful contractions at high velocity. In conclusion, ACTN3 R577X polymorphism was associated with power athlete status in Russians.


Ethanol inhibits mitogen activated protein kinase activity and growth of vascular smooth muscle cells in vitro.
The aim of this study was to determine the effect of ethanol on vascular smooth muscle cell proliferation and mitogen activated protein kinase (MAPK) signaling. Rat aortic smooth muscle cell growth in vitro was determined by measuring cell counts and [3H]thymidine incorporation. MAPK signaling was determined by assessing MEK (also referred to as MAPK kinase) activity by measuring phosphorylated extracellular signal-regulated kinase (pp44ERK - 1 and pp42ERK - 2) expression, and ERK activity by measuring ERK-2-dependent phosphorylation of myelin basic protein (MBP). In quiesced smooth muscle cells, ethanol treatment (24 h) inhibited serum-stimulated mitogenesis in a dose-dependent manner, (IC50 = 60 mM), in the absence of any effect on smooth muscle cell viability. In addition, ethanol treatment caused a significant shift to the right in the smooth muscle cell growth curve, extending the population doubling time from approximately 48 h (control) to approximately 70 h (ethanol). Acute (15 min) ethanol treatment reduced serum-stimulated pp44ERK - 1 and pp42ERK - 2 expression in a dose dependent fashion; 24.5+/-1.5% and 77.6+/-3.2% inhibition for 20 mM and 160 mM ethanol, respectively. Furthermore, there was a significant dose-dependent decrease in ERK2 activity in ethanol treated smooth muscle cells as compared to control smooth muscle cells. These data demonstrate an inhibitory effect of ethanol on smooth muscle cell proliferation and MAPK signalling in vitro. It is tempting to speculate that these actions of ethanol may contribute to its cardiovascular effects in vivo.


Antigen-specific B-cell function in human autoimmune thyroiditis.
T-B cells from the peripheral circulation of patients with autoimmune thyroiditis were cocultured with sheep red blood cells (SRBC) or soluble human thyroglobulin (Tg), a self-antigen. The B-cell mitogen, Staphylococcus aureus, combined with macrophage-derived B-cell differentiating factor, induced in vitro lymphoid activation and proliferation in the presence or absence of Tg or SRBC, which was monitored after 6 days by specific anti-Tg and anti-SRBC plaque-forming cell (PFC) responses (expressed as PFC per 10(6) T-B cells). In the presence of SRBC, significantly more anti-SRBC PFC (322 +/- 113, SE) were generated in normals (N = 5) compared with autoimmune thyroiditis patients (58 +/- 36) (N = 8) (P less than or equal to 0.001), data consistent with an antigen-specific T-cell defect. Anti-Tg PFC, not detectable in normal controls, were observed in patients in the absence (111 +/- 41) and presence (171 +/- 64) of Tg (10-1000 ng/ml). However, variable responses were noted after such coculture experiments with Tg. Three patients demonstrated amplification of anti-Tg PFC, while three showed antigen-related inhibition of anti-Tg PFC. These data indicated heterogeneity of responses to Tg antigen in patients with autoimmune thyroid disease compounded by significantly depressed antigen-specific induction mechanisms.


Malaria and urbanization in central Africa: the example of Brazzaville. Part I: Description of the town and review of previous surveys.
For a growing number of citizens, the upheavals caused by rapid urban development completely transform malaria epidemiology in tropical Africa. This paper is the first of a series on this phenomenon in Brazzaville, capital of the Congo Republic, where a third of the country's population is already concentrated. After describing the main human and physical geographical aspects of the town, the most significant results of the previous studies of this town and the surrounding rural area are reviewed. They show the existence, until the beginning of the 1950s of a stable, holoendemic situation characterized by a malaria prevalence approaching 90% in children in both urban and rural areas. Since then the intensive development of anti-malaria campaigns in urban areas over about ten years led temporarily to a considerable decrease in the level of endemicity, while in rural areas it remained unchanged. In the 1960s, the vector control and systematic chemoprophylactic measures gradually stopped, permitting the study of a new malaria dynamic related only to the evolving particularities of a changing urban ecosystem.


Acoustic reflectometry esophageal profiles minimally affected by massive gas ventilation.
Acoustic reflectometry can be used to distinguish between breathing tube placement in an esophagus vs the trachea via characteristic area-distance profiles for both cavities. In the cardiopulmonary resuscitation setting, capnography may be useless because the patient has little or no pulmonary circulation. With the breathing tube in the esophagus, can massive ventilation with a manual resuscitation bag, as might occur in the cardiopulmonary resuscitation setting, markedly alter the form of the obtained esophageal reflectometry profile? Nine hounds were induced, endotracheally intubated, mechanically ventilated, and anesthetized. Area-distance profiles were obtained with a 2-microphone acoustic reflectometer customized to measure areas up to 50 cm. Acoustic reflectometer profiles were obtained in intubated esophagi as follows: (1) baseline nonventilated state, (2) after aggressive 2-handed manual ventilation with high inspiratory pressures, rapid respiratory rates, and large tidal volumes for periods of 0.5, 1, and 1.5 minutes, upon detachment of the resuscitation bag, and (3) after esophagogastric decompression. We hypothesized that massive gas ventilation has no effect on the esophageal peak areas (null hypothesis), and used a paired t test for statistical significance (P < .05). For times of 0.5, 1.0, and 1.5 minutes, the ventilation volumes (mean +/- SD) were 25 +/- 7, 49 +/- 8, and 70 +/- 18 L. Massive gas ventilation caused minimal broadening and slight distal spread of the basal "hump". The mean peak area change was 0.18 +/- 0.35 cm2. For a paired t test (n = 9, df = 8), the corresponding t value was 1.54, with a P value of .16, which was incompatible with the null hypothesis. The experimental observations indicate a minimal effect of massive gas ventilation on the acoustic reflectometry esophageal profile. Hence, operator recognition of the altered canine acoustic reflectometer profile as that of an esophageal cavity is maintained, indicating that acoustic reflectometry may be useful in correctly identifying the site of breathing tube placement in out-of-hospital cardiac arrest situations despite massive esophageal ventilation.


Adequate Pelvic Lymph Node Dissection in Radical Cystectomy in the Era of Neoadjuvant Chemotherapy: A Meta-Analysis and Systematic Review.
Radical cystectomy (RC) with pelvic lymphadenectomy (PLND) serves as the gold-standard treatment for muscle-invasive bladder cancer (MIBC). Numerous studies have shown that the number of lymph nodes (LN) removed during RC could affect patient prognosis. However, these studies confirmed the association between PLND and survival outcomes prior to the widespread adoption of neoadjuvant chemotherapy (NAC). Consequently, this study aimed to investigate the prognostic role of PLND in patients previously pretreated with NAC. A systematic review and meta-analysis were performed using PubMed, Web of Knowledge, and Scopus databases. The selected studies contained a total of 17,421 participants. The meta-analysis indicated a significant correlation between adequate PLND and overall survival in the non-NAC group. However, a survival benefit was not observed in patients undergoing RC with preoperative systemic therapy, regardless of the LN cut-off thresholds. The pooled HR for ≥10 and ≥15 LN were 0.87 (95% CI 0.75-1.01) and 0.87 (95% CI 0.76-1.00), respectively. The study results suggest that NAC mitigates the therapeutic significance of PLND, as patients pre-treated with NAC no longer gain oncological benefits from more extensive lymphadenectomy. This highlights the analogous roles of NAC and PLND in eradication of micrometastases and in prevention of distal recurrence post-RC.


Gap junctions in development--a perspective.
The status of the gap junction as a pathway for cellular interactions during development is reviewed. Current evidence suggests that gap junctions play an important part in ensuring normal development, although the precise role of gap junctional communication remains to be defined. Communication through gap junctions acts alongside cellular interactions achieved by the release of growth factors during embryogenesis. Differences between groups of developing cells may be reflected in, and possibly controlled by, alterations in the selectivity of the gap junctions. It seems likely that gap junctional communication is involved in the control of embryonic patterning rather than phenotypic differentiation.


Self-assembling tryptophan-based designer peptides as intracellular delivery vehicles.
A series of tryptophan-based peptides W1a, b-W4a, b, with diverse architectures were designed and synthesized. These tryptophan containing peptides can self-assemble to spherical particle. This self-assembled system was demonstrated to encapsulate rhodamine B and penetrate the cell membrane.


Pathogenetic mechanisms in combined cilioretinal artery and retinal vein occlusion: a reappraisal.
We examined nine patients who presented cilioretinal artery occlusion (CLRAO) associated with retinal vein occlusion (RVO). CLRAO was probably secondary to the raised intraluminal resistance consequent to the RVO in patients showing initially a delayed filling of the cilioretinal artery in fluorescein angiography. Interestingly, these patients presented an ophthalmoscopically more severe form of RVO and had systemic predisposing factors for a RVO. In patients presenting a physiological perfusion of the cilioretinal artery in fluorescein angiography, RVO was a self limited disease and etiologic factors were not found. This may suggest that in these patients the CLRAO probably occurred simultaneously with the RVO after a decrease in perfusion pressure in both retinal and cilioretinal arterial systems. In this combined vaso-occlusive retinopathy the vulnerability of cilioretinal arteries can be explained either by the absence of autoregulation or by their lower perfusion pressure gradient in comparison with retinal arteries.


Inflammatory cytokines in newborn infants.
Serum levels of IL-1beta, IL-6 and TNF-alpha were measured in 48 healthy, termed neonates on the 1st (N1), 5th (N5) and 40th (N40) day after birth, compared with those in maternal serum (MS), umbilical cord (UC) and adult controls. Cytokine values in N1 and N5 were significantly elevated, than those in UC and in controls (P<0.0001). IL-1beta and IL-6 declined significantly from N1 to N40 (P<0.0001), while TNF-alpha increased significantly from N1 to N5 and declined thereafter. MS infinity IL-1beta and IL-6, but not MS infinity TNF-alpha, were significantly higher than those of controls (P<0.0001). IL-1beta values depended on the mode of delivery. In conclusion, the increased concentrations of IL-1beta, IL-6 and TNF-alpha during the perinatal period might suggest their involvement in an inflammation-like process during normal parturition, and reflect also a newborn immune response to the stress of delivery and environmental changes.


Sunflower seeds classification based on sparse convolutional neural networks in multi-objective scene.
Generally, sunflower seeds are classified by machine vision-based methods in production, which include using photoelectric sensors to identify light-sensitive signals through traditional algorithms for which the equipment cost is relatively high and using neural network image recognition methods to identify images through cameras for which the computational cost is high. To address these problems, a multi-objective sunflower seed classification method based on sparse convolutional neural networks is proposed. Sunflower seeds were obtained from the video recorded using the YOLOv5 Object detection algorithm, and a ResNet-based classification model was used to classify the seeds according to differences in appearance. The ResNet has the disadvantages of having numerous parameters and high storage requirements; therefore, this study referred to the Lottery Ticket Hypothesis and used the Iterative Magnitude Pruning algorithm to compress the sunflower seed classification model, aiming to ascertain the optimal sparse sub-network from the classification model. Experiments were conducted to compare the effects on model performance before and after pruning, pruning degree, and different pruning methods. The results showed that the performance of the ResNet-based sunflower seed classification model using global pruning was the least affected by pruning, with a 92% reduction in the number of parameters, the best accuracy is 0.56% better than non-pruned and 9.17% better than layer-wise pruning. These findings demonstrate that using the Iterative Magnitude Pruning algorithm can render the sunflower seed classification model lightweight with less performance loss. The reduction in computational resources through model compression reduces the cost of sunflower seed classification, making it more applicable to practical production, and this model can be used as a cost-effective alternative to key sunflower seed classification techniques in practical production.


Imaging diagnosis and minimally-invasive management of necrotizing fasciitis in a dog.
Necrotising fasciitis is a rapidly progressive, aggressive bacterial infection of the subcutis associated with significant morbidity and mortality in both man and domestic animals. To the best of our knowledge, this is the first veterinary report of magnetic resonance imaging findings in necrotising fasciitis, and the first reported case in a dog to be successfully treated with minimally invasive surgical intervention.


Pharmacokinetics of clearance in the maternal-fetal amniotic fluid system of the rat.
A pharmacokinetic model of the maternal-fetal amniotic fluid system was designed to permit calculation of clearances from the steady-state concentrations of p-phenylbenzoic acid (PPBA) during infusion of PPBA into the maternal vein and the amniotic fluid of pregnant rats on the 16th to the 21st day of gestation. Transplacental clearance from the fetus to the mother maintained fairly constant values until the 19th day of gestation but decreased on Days 20 and 21 of gestation. Both clearances from the fetus into the amniotic fluid and from the amniotic fluid into the fetus showed higher values during the earlier stages of gestation, indicating that amniotic fluid may play an important role in the pharmacokinetics of PPBA in the fetus during the earlier stage of gestation. Fetal tissue clearance appeared to contribute to the elimination of PPBA in the fetus on Days 20 and 21 of gestation. Transplacental clearance from the mother to the fetus and clearance by nonplacental elimination in the mother increased with the day of gestation. The relationship between developmental changes in clearances and physiological changes in placenta and fetal tissues is discussed. The present model provides a basis for calculation of clearances in the maternal-fetal amniotic fluid system.


Optimizing immunofluorescence for the testing of autoantibodies in inner ear disorders.
A method is described for the immunofluorescent testing of autoantibodies in the sera of patients with vestibulo-cochlear disorders, using fixed decalcified inner ear tissue preparations from guinea pigs. Fixation in cold ethanol and decalcification in EDTa at 4 degrees C preserved the immunological reactivity of the inner ear tissue in use and allowed excellent delineation of its structural details. Counterstaining of the sections with Evans-blue dye aided in a better identification of the inner ear anatomical structures and in a more precise location of the antibodies present. By mounting the sections in p-phenylenediamine-PBS-glycerin solution, the rapid extinction of the fluorescence seen during photomicroscopy was retarded and the intensity of the immunofluorescence was enhanced. Preservation of the slides for documentation was improved by fixing the intermediate layer of the antigen and the labeled antibody in cold ethanol. The inner ear tissue preparations retained their fluorescent staining up to 6 months. Our method is convenient for screening patients with inner ear disorders for autoantibodies against the different cellular elements and for testing the possible presence of antibodies against the inner ear tissue. So far, we believe that the antibodies detected are not tissue (inner ear) specific.


Treatment of acute diarrhea.
The application of new pathophysiologic information to the treatment of diarrheal diseases has greatly enhanced the outlook for millions of young patients in many developing countries. This article outlines the basis and nature of these therapeutic advances in the belief that they are relevant, not only in Asia and South America, but also throughout North America.


Structural and evolutionary insights into the multidomain galectin from the red abalone Haliotis rufescens with specificity for sulfated glycans.
Galectins are an evolutionarily ancient family of lectins characterized by their affinity for β-galactosides and a conserved binding site in the carbohydrate recognition domain (CRD). These lectins are involved in multiple physiological functions, including the recognition of glycans on the surface of viruses and bacteria. This feature supports their role in innate immune responses in marine mollusks. Here, we identified and characterized a galectin, from the mollusk Haliotis rufescens (named HrGal), with four CRDs that belong to the tandem-repeat type. HrGal was purified by affinity chromatography in a galactose-agarose resin and exhibited a molecular mass of 64.11 kDa determined by MALDI-TOF mass spectrometry. The identity of HrGal was verified by sequencing, confirming that it is a 555 amino acid protein with a mass of 63.86 kDa. This protein corresponds to a galectin reported in GenBank with accession number AHX26603. HrGal is stable in the presence of urea, reducing agents, and ions such as Cu2+ and Zn2+. The recombinant galectin (rHrGal) was purified from inclusion bodies in the presence of these ions. A theoretical model obtained with the AlphaFold server exhibits four non-identical CRDs, with a β sandwich folding and the representative motifs for binding β-galactosides. This allows us to classify HrGal within the tandem repeat galectin family. On the basis of a phylogenetic analysis, we found that the mollusk sequences form a monophyletic group of tetradomain galectins unrelated to vertebrate galectins. HrGal showed specificity for galactosides and glucosides but only the sulfated sugars heparin and ι-carrageenan inhibited its hemagglutinating activity with a minimum inhibitory concentration of 4 mM and 6.25 X 10-5% respectively. The position of the sulfate groups seemed crucial for binding, both by carrageenans and heparin.


Constitutive androstane receptor regulates the intestinal mucosal response to injury.
The pathogenesis of the inflammatory bowel diseases (IBD), comprising Crohn's disease (CD) and ulcerative colitis (UC), involves aberrant interactions between a genetically susceptible individual, their microbiota and environmental factors. Alterations in xenobiotic receptor expression and function are associated with increased risk for IBD. Here, we have assessed the role of the constitutive androstane receptor (CAR), a xenobiotic receptor closely related to the pregnane X receptor, in the regulation of intestinal mucosal homeostasis. CAR expression was assessed in intestinal mucosal biopsies obtained from CD and UC patients, and in C57/Bl6 mice exposed to dextran sulphate sodium (DSS; 3.5% w/v in drinking water) to evoke intestinal inflammation and tissue damage. CAR-deficient mice were exposed to DSS and mucosal healing assessed. Modulation of wound healing by CAR was assessed in vitro. The therapeutic potential of CAR activation was evaluated, using 3,3',5,5'-tetrachloro-1,4-bis(pyridyloxy)benzene (TCPOBOP), a selective rodent CAR agonist. CAR expression was reduced in CD and UC samples, compared with expression in healthy controls. This was reproduced in our DSS studies, where CAR expression was reduced in colitic mice. CAR-deficient mice exhibited reduced healing following DSS exposure. In vitro, CAR activation accelerated intestinal epithelial wound healing by enhancing cell migration. Lastly, treating mice with TCPOBOP, following induction of colitis, enhanced mucosal healing. Our results support the notion that xenobiotic sensing is altered during intestinal inflammation, and suggest that CAR activation may prove effective in enhancing mucosal healing in patients with IBD.


Epidemiology and outcomes of pediatric burns over 35 years at Parkland Hospital.
Since opening its doors in 1962, the Parkland Burn Center has played an important role in improving the care of burned children through basic and clinical research while also sponsoring community prevention programs. The aim of our study was to retrospectively analyze the characteristics and outcomes of pediatric burns at a single institution over 35 years. The institutional burn database, which contains data from January 1974 until August 2010, was retrospectively reviewed. Patients older than 18 years of age were excluded. Patient age, cause of burn, total body surface area (TBSA), depth of burn, and patient outcomes were collected. Demographics were compared with regional census data. Over 35 years, 5748 pediatric patients were admitted with a thermal injury. Males comprised roughly two-thirds (66.2%) of admissions. Although the annual admission rate has risen, the incidence of pediatric burn admissions, particularly among Hispanic and African American children has declined. The most common causes of admission were scald (42%), flame (29%), and contact burns (10%). Both the median length of hospitalization and burn size have decreased over time (r(2)=0.75 and 0.62, respectively). Mortality was significantly correlated with inhalation injury, size of burn, and history of abuse. It was negatively correlated with year of admission. Over 35 years in North Texas, the median burn size and incidence of pediatric burn admissions has decreased. Concomitantly, length of stay and mortality have also decreased.


The molecular size and shape of xanthan, xylinan, bronchial mucin, alginate, and amylose as revealed by electron microscopy.
Electron microscopy of some selected, vacuum-dried and rotary-shadowed, polyelectrolytic polysaccharides and glycoproteins adsorbed to mica indicates that this technique can yield reliable information about polymer conformation for chains with persistence lengths q exceeding about 10 nm. Statistical analyses of the local polymer tangent-direction yield q = 150 nm for double-stranded xanthan, q = 60 nm for single-stranded xanthan, q = 45 nm for xylinan, q = 16 nm for alginate (90% beta-D-mannuronic acid), and q = 15 nm for human-bronchial mucin. These values are all in adequate agreement with values of q obtained by using other techniques. Amylose, on the other hand, appears as non-randomly aligned chains. The observed contour lengths of amylose indicate a mass per unit length of 1440 dalton/nm, consistent with a pseudo-helical conformation.


Diamine-modified porous indium frameworks with crystalline porous materials (CPM)-5 structure for carbon dioxide fixation under co-catalyst and solvent free conditions.
In the present work, functional diamine groups into indium frameworks to synthesize cyclic carbonates from CO2 and epoxides with efficient catalytic activity in the absence of co-catalyst and solvent are reported for the first time. Crystalline porous materials (CPM)-5 modified with 1,2-phenylene diamine and ethylene diamine (CPM-5-PhDA and CPM-5-EDA), were prepared using a post-synthetic modification (PSM) method. The properties of the modified CPM-5 were characterized by powder X-ray diffraction (PXRD), Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), N2-adsorption, scanning electron microscopy (SEM), CO2 adsorption, and temperature programmed desorption TPD methods. The presence of diamine groups as basic sites and indium Lewis acid sites in the framework structure were desirable for high catalytic activity. For a given catalyst weight, CPM-5-PhDA was the best candidate to appear with great catalytic activity and selectivity for the cycloaddition reaction at 100°C and 1 MPa CO2 under co-catalyst and solvent free conditions. CPM-5-PhDA also was found to afford large and bulky epoxides. The catalyst can be easily separated and reused five times without any decline in activity.


Establishment of an African green monkey model for COVID-19 and protection against re-infection.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for an unprecedented global pandemic of COVID-19. Animal models are urgently needed to study the pathogenesis of COVID-19 and to screen vaccines and treatments. We show that African green monkeys (AGMs) support robust SARS-CoV-2 replication and develop pronounced respiratory disease, which may more accurately reflect human COVID-19 cases than other nonhuman primate species. SARS-CoV-2 was detected in mucosal samples, including rectal swabs, as late as 15 days after exposure. Marked inflammation and coagulopathy in blood and tissues were prominent features. Transcriptome analysis demonstrated stimulation of interferon and interleukin-6 pathways in bronchoalveolar lavage samples and repression of natural killer cell- and T cell-associated transcripts in peripheral blood. Despite a slight waning in antibody titers after primary challenge, enhanced antibody and cellular responses contributed to rapid clearance after re-challenge with an identical strain. These data support the utility of AGM for studying COVID-19 pathogenesis and testing medical countermeasures.


Blood glucose and epicardial adipose tissue at the hospital admission as possible predictors for COVID-19 severity.
To study the possible association of CT-derived quantitative epicardial adipose tissue (EAT) and glycemia at the admission, with severe outcomes in patients with COVID-19. Two hundred and twenty-nine patients consecutively hospitalized for COVID-19 from March 1st to June 30th 2020 were studied. Non contrast chest CT scans, to confirm diagnosis of pneumonia, were performed. EAT volume (cm3) and attenuation (Hounsfield units) were measured using a CT post-processing software. The primary outcome was acute respiratory distress syndrome (ARDS) or in-hospital death. The primary outcome occurred in 56.8% patients. Fasting blood glucose was significantly higher in the group ARDS/death than in the group with better prognosis [114 (98-144) vs. 101 (91-118) mg/dl, p = 0.001]. EAT volume was higher in patients with vs without the primary outcome [103 (69.25; 129.75) vs. 78.95 (50.7; 100.25) cm3, p < 0.001] and it was positively correlated with glycemia, PCR, fibrinogen, P/F ratio. In the multivariable logistic regression analysis, age and EAT volume were independently associated with ARDS/death. Glycemia and EAT attenuation would appear to be factors involved in ARDS/death with a trend of statistical significance. Our findings suggest that both blood glucose and EAT, easily measurable and modifiable targets, could be important predisposing factors for severe Covid-19 complications.


Cyclin-Dependent Kinase Inhibitor 2A is a Key Regulator of Cell Cycle Arrest and Senescence in Endothelial Colony-Forming Cells in Moyamoya Disease.
Endothelial colony-forming cells (ECFCs) have been reported to play an important role in the pathogenesis of moyamoya disease (MMD). We have previously observed stagnant growth in MMD ECFCs with functional impairment of tubule formation. We aimed to verify the key regulators and related signaling pathways involved in the functional defects of MMD ECFCs. ECFCs were cultured from peripheral blood mononuclear cells of healthy volunteers (normal) and MMD patients. Low-density lipoproteins uptake, flow cytometry, high content screening, senescence-associated β-galactosidase, immunofluorescence, cell cycle, tubule formation, microarray, real-time quantitative polymerase chain reaction, small interfering RNA transfection, and western blot analyses were performed. The acquisition of cells that can be cultured for a long time with the characteristics of late ECFCs was significantly lower in the MMD patients than the normal. Importantly, the MMD ECFCs showed decreased cellular proliferation with G1 cell cycle arrest and cellular senescence compared to the normal ECFCs. A pathway enrichment analysis demonstrated that the cell cycle pathway was the major enriched pathway, which is consistent with the results of the functional analysis of ECFCs. Among the genes associated with the cell cycle, cyclin-dependent kinase inhibitor 2A (CDKN2A) showed the highest expression in MMD ECFCs. Knockdown of CDKN2A in MMD ECFCs enhanced proliferation by reducing G1 cell cycle arrest and inhibiting senescence through the regulation of CDK4 and phospho retinoblastoma protein. Our study suggests that CDKN2A plays an important role in the growth retardation of MMD ECFCs by inducing cell cycle arrest and senescence.


Sequence Learning in Minimally Verbal Children With ASD and the Beneficial Effect of Vestibular Stimulation.
People with autism spectrum disorder (ASD) and especially the minimally verbal, often fail to learn basic perceptual and motor skills. This deficit has been demonstrated in several studies, but the findings could have been due to the nonoptimal adaptation of the paradigms. In the current study, we sought to characterize the skill learning deficit in young minimally verbal children with ASD and explore ways for improvement. For this purpose, we used vestibular stimulation (VS) whose beneficial effects have been demonstrated in the typical population, but the data regarding ASD are limited. We trained 36 children ages 6-13 years, ASD (N = 18, 15 of them minimally verbal) and typical development (TD, N = 18), on a touch version of the visual-motor Serial-Reaction-Time sequence-learning task, in 10 short (few minutes) weekly practice sessions. A subgroup of children received VS prior to each training block. All the participants but two ASD children showed gradual median reaction time improvement with significant speed gains across the training period. The ASD children were overall slower (by ~250 msec). Importantly, those who received VS (n = 10) showed speed gains comparable to TD, which were larger (by ~100%) than the ASD controls, and partially sequence-specific. VS had no effect on the TD group. These results suggest that VS has a positive effect on learning in minimally verbal ASD children, which may have important therapeutic implications. Furthermore, contrary to some previous findings, minimally verbal children with ASD can acquire, in optimal conditions, procedural skills with few short training sessions, spread over weeks, and with a similar time course as non-ASD controls. Autism Res 2020, 13: 320-337. © 2019 International Society for Autism Research, Wiley Periodicals, Inc. LAY SUMMARY: Minimally verbal children with ASD who received specially adjusted learning conditions showed significant learning of a visual-motor sequence across 10 practice days. This learning was considerably improved with vestibular stimulation before each short learning session. This may have important practical implications in the education and treatment of ASD children.


Sodium accumulation in the skin is associated with higher density of skin lymphatic vessels in patients with arterial hypertension.
Recent studies, conducted mainly on the rodent model, have demonstrated that regulatory pathway in the skin provided by glycosaminoglycans, nuclear factor of activated T cells 5 (NFAT5), vascular endothelial growth factor C (VEGF-C) and process of lymphangiogenesis may play an important role in extrarenal regulation of sodium (Na+) balance, body water volume, and blood pressure. We aimed to investigate the concentrations and relations among the main factors of this pathway in human skin to confirm that this regulatory axis also exists in humans. Skin specimens from patients diagnosed with arterial hypertension and from control group were histologically and molecularly examined. The primary hypertensive and control groups did not differ in Na+ concentrations in the skin. However, the patients with hypertension and higher skin Na+ concentration had significantly greater density of skin lymphatic vessels. Higher skin Na+concentration was associated with higher skin water content. In turn, skin water content correlated with factors associated with lymphangiogenesis, i.e. NFAT5, VEGF-C, and podoplanin (PDPN) mRNA expression in the skin. The strong mutual pairwise correlations of the expressions of NFAT5, VEGF-C, vascular endothelial growth factor D (VEGF-D) and PDPN mRNA were noted in the skin in all of the studied groups. Our study confirms that skin interstitium and the lymphatic system may be important players in the pathophysiology of arterial hypertension in humans. Based on the results of our study and existing literature in this field, we propose the hypothetical model which might explain the phenomenon of salt-sensitivity.


Secretory lysosomes.
Regulated secretion of stored secretory products is important in many cell types. In contrast to professional secretory cells, which store their secretory products in specialized secretory granules, some secretory cells store their secretory proteins in a dual-function organelle, called a secretory lysosome. Functionally, secretory lysosomes are unusual in that they serve both as a degradative and as a secretory compartment. Recent work shows that cells with secretory lysosomes use new sorting and secretory pathways. The importance of these organelles is highlighted by several genetic diseases, in which immune function and pigmentation--two processes that normally involve secretory lysosomes--are impaired.


Modification of egg production and packaging in response to food availability by Nassarius pauperatus.
Adjacent populations of Nassarius pauperatus differ significantly in how hungry their members are. The numbers of eggs and egg capsules produced are positively correlated with how hungry members of a population are, but the average number of eggs per capsule is negatively correlated with hunger. These results were confirmed in a laboratory experiment using two groups from the same population maintained with different levels of food.The significance of this behaviour is discussed and the hypothesis proposed that the flexibility of reproductive patterns is correlated with dispersive ability.


Myelodysplasia in cadaver renal allografts: a report of four cases.
A report of four cases of severe myelodysplastic syndromes (MDS) culminating in death in cadaver renal recipients is presented. All patients received predominantly antimetabolite immunosuppressive therapy. Abnormal cytogenetics were found in two cases. Survival was brief in all cases from the time of diagnosis. These cases add to the increasing number of patients treated with antimetabolite drugs for nonmalignant conditions who develop severe hematological abnormalities which might be a prodrome of acute nonlymphocytic leukemia.


Effect of magnesium and polyamines on the structure of yeast tRNAPhe.
The effect of magnesium and polyamines (spermine, spermidine, putrescine and cadaverine) on the structure of yeast tRNAPhe has been investigated. It is found that magnesium induces structural changes and stabilizes hydrogen bonds in the temperature range 22--44 degrees C in 0.17 M sodium. The number of Mg2+ which affect tRNA structure increases from 1 +/- 1 at 22 degrees C to 4 +/- 1 at 44 degrees C and the number of additional base pairs formed in the presence of magnesium increases from 1 +/- 1 at 22 degrees C to 4 +/- 1 at 44 degrees C. The spectral changes are more-or-less sequential. The polyamine spermine stabilizes some, but not all, of the structural features stabilized by magnesium at 44 degrees C, and the combination of magnesium and spermine, at low levels, is more effective than either cation alone in stabilizing tRNA structure. Comparison of the effects of spermine, spermidine, putrescine and cadaverine indicates that it is the asymmetric triamine unit which is important in the stabilization. Some spectral changes induced by magnesium can be assigned to stabilization of specific tertiary structure interactions and to alteration of stacking adjacent to U8-A14.


An event based topic learning pipeline for neuroimaging literature mining.
Neuroimaging text mining extracts knowledge from neuroimaging texts and has received widespread attention. Topic learning is an important research focus of neuroimaging text mining. However, current neuroimaging topic learning researches mainly used traditional probability topic models to extract topics from literature and cannot obtain high-quality neuroimaging topics. The existing topic learning methods also cannot meet the requirements of topic learning oriented to full-text neuroimaging literature. In this paper, three types of neuroimaging research topic events are defined to describe the process and result of neuroimaging researches. An event based topic learning pipeline, called neuroimaging Event-BTM, is proposed to realize topic learning from full-text neuroimaging literature. The experimental results on the PLoS One data set show that the accuracy and completeness of the proposed method are significantly better than the existing main topic learning methods.


Diverse mutational pathways converge on saturable chloroquine transport via the malaria parasite's chloroquine resistance transporter.
Mutations in the chloroquine resistance transporter (PfCRT) are the primary determinant of chloroquine (CQ) resistance in the malaria parasite Plasmodium falciparum. A number of distinct PfCRT haplotypes, containing between 4 and 10 mutations, have given rise to CQ resistance in different parts of the world. Here we present a detailed molecular analysis of the number of mutations (and the order of addition) required to confer CQ transport activity upon the PfCRT as well as a kinetic characterization of diverse forms of PfCRT. We measured the ability of more than 100 variants of PfCRT to transport CQ when expressed at the surface of Xenopus laevis oocytes. Multiple mutational pathways led to saturable CQ transport via PfCRT, but these could be separated into two main lineages. Moreover, the attainment of full activity followed a rigid process in which mutations had to be added in a specific order to avoid reductions in CQ transport activity. A minimum of two mutations sufficed for (low) CQ transport activity, and as few as four conferred full activity. The finding that diverse PfCRT variants are all limited in their capacity to transport CQ suggests that resistance could be overcome by reoptimizing the CQ dosage.


Purification and characterization of l-glutaminase enzyme from camel liver: Enzymatic anticancer property.
l-Glutaminase has gained an important attention as glutamine-depleting enzyme in treatment of various cancers. Therefore, this study aimed to purify, characterize and investigate antitumor activity of l-glutaminase from camel liver mitochondria (CL-Glu), since no available information about CL-Glu from camel. CL-Glu was purified using cell fractionation, ultrafiltration, DEAE-and CM-cellulose chromatography columns. The purified CL-Glu was a monomer with a molecular weight of 70 ± 3 kDa, isoelectric point of 7.2, optimum temperature of 70 °C and it was active over a broad pH range with a pH optimum at pH 8.0. Its activity had a clear dependence on phosphate ions. The studied enzyme showed sigmoidal kinetics, indicated its allosteric behavior with Km of 36 ± 4 mM and Hill coefficient of 1.5 which suggested a positive cooperatively of active sites. The purified l-glutaminase exerted antitumor activity against different cell lines with the highest cytotoxic activity against Hepatocellular carcinoma cell line (HepG-2) with an IC50 value of 152 µg/ml. In conclusion, l-glutaminase was purified from camel liver using simple methods and its unique properties such as stability at both wide pH range and at high temperature along with its relatively low molecular weight, facilitated its usage in medical applications as antitumor drug.


Role of NMDAR plasticity in a computational model of synaptic memory.
A largely unexplored question in neuronal plasticity is whether synapses are capable of encoding and learning the timing of synaptic inputs. We address this question in a computational model of synaptic input time difference learning (SITDL), where N-methyl-d-aspartate receptor (NMDAR) isoform expression in silent synapses is affected by time differences between glutamate and voltage signals. We suggest that differences between NMDARs' glutamate and voltage gate conductances induce modifications of the synapse's NMDAR isoform population, consequently changing the timing of synaptic response. NMDAR expression at individual synapses can encode the precise time difference between signals. Thus, SITDL enables the learning and reconstruction of signals across multiple synapses of a single neuron. In addition to plausibly predicting the roles of NMDARs in synaptic plasticity, SITDL can be usefully applied in artificial neural network models.


Risk of incidental finding of endometrial cancer at the time of hysterectomy for benign condition.
To assess the risk of unanticipated endometrial carcinoma during hysterectomy to treat a presumed benign condition. Retrospective analysis (Canadian Task Force classification II-2). Tertiary referral center, university hospital. All women who underwent hysterectomy to treat presumed benign indications at our center from January 2000 to December 2011 were identified. We analyzed all pathologic reports and identified cases of unexpected endometrial carcinoma. Hysterectomy by any approach to treat presumed benign indications. At our institution, 2179 hysterectomies were performed to treat presumed benign indications. Nine (0.4%) revealed unexpected endometrial carcinoma (95% confidence interval, 0.2-0.7). Our data suggest that the rate of unanticipated endometrial carcinoma during hysterectomy to treat benign conditions is low.


Application of a Markov chain Monte Carlo method for robust quantification in chemical exchange saturation transfer magnetic resonance imaging.
Chemical exchange saturation transfer (CEST) magnetic resonance imaging can provide surrogate biomarkers for disease diagnosis. However, endogenous CEST effects are always diluted and contaminated by competing effects, which results in unwanted signal contributions that lessen the specificity of CEST to underlying biochemical exchange processes. The aim of this study was to examine a method for the accurate quantification of CEST effects. A Markov chain Monte Carlo (MCMC)-based Bayesian inference approach was proposed to estimate the exchange parameters, and CEST effects could be fitted using these estimations. This approach was tested in Bloch simulation and ischemic stroke rat experiments, and its performance was evaluated using quantification maps and numerical metrics. With 12 groups of simulations, the MCMC method achieved satisfactory fittings on both 2-pool and 5-pool models. The sum of squares error values and the root mean square error of the fitted Z-spectra were smaller than 10-3, and the coefficient of determination (R-squared) values were close to 1. The corresponding CEST quantification M T R R e x C E S T spectra were also well fitted and successfully separated the mixed CEST effects. The estimated parameters showed little bias relative to the ground truth, with errors between the true and estimated values of each parameter of less than 0.5%. In the animal experiments, M T R R e x A m i d e fitted using the MCMC method showed obvious contrast between ischemic and contralateral regions at the early stage. Compared with other quantification methods, it displayed the highest contrast-to-noise ratios (3.9, 2.73, and 3.93) and the lowest coefficient of variation values (0.181, 0.2224, and 0.2897) in all three stroke periods. The MCMC method provided an efficient approach for parameter estimation and CEST effect quantification. The method may therefore be useful in achieving an accurate pathological diagnosis.


The Toolbox for Modified Aptamers.
Aptamers are nucleic acid-based scaffolds that can bind with high affinity to a variety of biological targets. Aptamers are identified from large DNA or RNA libraries through a process of directed molecular evolution (SELEX). Chemical modification of nucleic acids considerably increases the functional and structural diversity of aptamer libraries and substantially increases the affinity of the aptamers. Additionally, modified aptamers exhibit much greater resistance to biodegradation. The evolutionary selection of modified aptamers is conditioned by the possibility of the enzymatic synthesis and replication of non-natural nucleic acids. Wild-type or mutant polymerases and their non-natural nucleotide substrates that can support SELEX are highlighted in the present review. A focus is made on the efforts to find the most suitable type of nucleotide modifications and the engineering of new polymerases. Post-SELEX modification as a complementary method will be briefly considered as well.


Preimplantation genetic screening: results of a worldwide web-based survey.
Our objective was to evaluate and characterize the extent and patterns of worldwide usage of preimplantation genetic screening (PGS) among the assisted reproductive technique community. A prospective, web-based questionnaire with questions relating to practices of, and views on, PGS was directed to users and non-users of PGS. A total of 386 IVF units from 70 countries conducting 342,600 IVF cycles annually responded to the survey. A total of 77% of respondents routinely carry out PGS in their clinics for a variety of indications: advanced maternal age (27%), recurrent implantation failure (32%) and recurrent pregnancy loss (31%). Few (6%) offer PGS to all their patients. In most cycles (72%), trophectoderm biopsy is carried out and either array-comparative genomic hybridization (59%) or next-generation sequencing (16%) are used for genetic analysis. Only 30% of respondents regard PGS as clearly evidenced-based, and most (84%) believe that more randomized controlled trials are needed to support the use of PGS. Despite ongoing debate and lack of robust evidence, most respondents support the use of PGS, and believe that it may aid in transferring only euploid embryos, thereby reducing miscarriage rates and multiple pregnancies, increasing live birth rates and reducing the risk of aneuploid pregnancies and births.