In vitro evaluation of the effects of Ultrasound Tongue Scraper on bacteria and biofilm formation.
Oral malodor is a common condition caused by some Gram-negative oral bacteria, among which are the 3 red complex bacteria (RCB). The present study investigated the effectiveness of the Ultrasound Tongue Scraper (UTS) to disrupt the structural morphology of the bacteria and their biofilm. While developing over 72 hours, multispecies biofilms of RCB (Porphromonas gingivalis, Tryponema denticola, Tannerella forsythia) were treated every 24 hours with 1.6-MHz ultrasound waves generated with UTS. An untreated group served as controls. Confocal laser scanning microscopy was used to determine the biofilm thickness, biomass and live : dead cell ratio at each time point (24, 48 and 72 hours). Biofilm morphology and bacteria ultrastructure were viewed using scanning/transmission electron microscopy, respectively. Data were analyzed using ANOVA and Tukey tests. At each time point, the 3 variables were significantly lower in treated samples than the untreated. Significant biofilm disruption was observed in treated samples at each time period while the untreated had intact biofilm morphology. Cells in treated samples showed disrupted cell wall, cytoplasmic material, huge vacuoles and heterogeneity in electron density, while these cell organelles remained intact in untreated samples. The UTS has an inhibitory effect on RCB and could be useful for oral malodor management.


Early enteral feeding and nosocomial sepsis in very low birthweight infants.
The interrelations between early enteral feeding, necrotising enterocolitis (NEC), and nosocomial sepsis (NS) remain unclear. To evaluate the effect of age at the introduction of enteral feeding on the incidence of NS and NEC in very low birthweight (VLBW< 1500 g) infants. Data were collected on the pattern of enteral feeding and perinatal and neonatal morbidity on all VLBW infants born in one centre during 1995-2001. Enteral feeding was compared between infants with and without NS and/or NEC. The study sample included 385 infants. Of these, 163 (42%) developed NS and 35 (9%) developed NEC. Enteral feeding was started at a significantly earlier mean (SD) age in infants who did not develop nosocomial sepsis (2.8 (2.6) v 4.8 (3.7) days, p = 0.0001). Enteral feeding was introduced at the same age in babies who did or did not develop NEC (3.1 (2) v 3.7 (3) days, p = 0.28). Over the study period, the mean annual age at the start of enteral feeding fell consistently, and this correlated with the mean annual incidence of NS (r(2) = 0.891, p = 0.007). Multiple logistic regression analysis showed age at start of enteral feeding, respiratory distress syndrome, and birth weight to be the most significant predictors of risk of NS (p = 0.0005, p = 0.024, p = 0.011). Early enteral feeding was associated with a reduced risk of NS but no change in the risk of NEC in VLBW infants. These findings support the use of early enteral feeding in this high risk population, but this needs to be confirmed in a large randomised controlled trial.


Effects of energy supplementation on energy losses and nitrogen balance of steers fed green-chopped wheat pasture I: Calorimetry.
Cattle grazing wheat pasture in the southern Great Plains are sometimes fed an energy supplement; however, the benefits of supplementation on nutrient balance, energy metabolism, and greenhouse gas emissions have not been elucidated. Therefore, we used 10 British crossbred steers (206 ± 10.7 kg initial BW) in a respiration calorimetry study to evaluate the effects of energy supplementation on energy losses, N balance, and nutrient digestibility of steers fed green-chopped wheat forage. The study design was an incomplete replicated 4 × 4 Latin square with treatments in a 2 × 2 factorial arrangement. Steers ( = 8) were assigned to 1 of 2 BW blocks (4 steers per block) with dietary factors consisting of 1) no supplementation (CON) or supplemented with a steam-flaked corn-based energy supplement (that also contained monensin sodium) at 0.5% of BW daily (SUP) and 2) NEm intakes of 1 times (1x) or 1.5 times (1.5x) maintenance. Wheat forage was harvested daily and continuously fed as green-chop to steers during the 56-d study. There were no differences ( ≥ 0.32) between CON and SUP for OM (78.3 vs. 80.7%, respectively) or NDF (68.3 vs. 64.8%, respectively) digestibility. At the 1.5x level of intake, there was no difference ( ≥ 0.16) in energy lost in feces (4.27 vs. 3.92 Mcal/d) or urine (0.58 vs. 0.55 Mcal/d), heat production (8.69 vs. 8.44 Mcal/d), or retained energy (3.10 vs. 3.46 Mcal/d) between supplementation treatments. Oxygen consumption (1,777 vs. 1,731 L/d; = 0.67) and CO production (1,704 vs. 1,627 L/d; = 0.56) of CON and SUP steers, respectively, were not different; however, SUP steers tended to have ( = 0.06) lower CH production (115 vs 130 L/d) than CON steers. Methane, as a proportion of GE intake, was similar for CON (6.87%) and SUP (6.07%; = 0.18), as was the ME:DE ratio ( = 0.24; 86.3% for CON and 87.9% for SUP). Fractional N excretion in urine and feces, as a proportion of total N excreted ( ≥ 0.84) or N intake ( ≥ 0.63), was not different between treatments. Calculated NEm and NEg values for CON were 1.76 and 1.37 Mcal/kg DM, respectively, whereas the NEm and NEg values for the SUP treatment were 2.32 and 1.61 Mcal/kg DM, respectively. Calculated NE values for steers fed additional energy were approximately 17.5% greater than the expected difference in energy content. This was probably the result of the inconsistent response at the 1x DMI level. Under these circumstances, energy supplementation did appear to enhance NEm and NEg value of the supplemented wheat forage diet.


[Vancomycin-resistant enterococci: in vitro activity of quinupristin / dalfoprostin (RP 59500)].
The recent emergence of glycopeptide-resistant enterococci limits the treatment of enterococcal infections. The aim of this study was to evaluate the in vitro activity of a new streptogramin, quinupristin/dalfopristin, against 30 clinical isolates of vancomycin-resistant enterococci and compared with those of other 15 antimicrobials. Enterococci were identified by using Rapid ID 32 Strep system. Genotyping of the isolates was performed by PCR. The MICs of quinupristin/dalfopristin were determined by the agar dilution technique recommended by the NCCLS. Susceptibilities to the rest of antibiotics tested (teicoplanin, ampicillin, penicillin, imipenem, doxycicline, chloramphenicol, gentamicin, streptomycin, rifampin, levofloxacin, fleroxacin, trovafloxacin, sparfloxacin, pefloxacin and clinafloxacin) were determined by using the E test. beta-lactamase production was examined with nitrocefin disks. Quinupristin/dalfopristin has demonstrated excellent activity against Enterococcus faecium (MIC90' 2 micrograms/ml). Enterococcus faecalis was considerably less susceptible than E. faecium, at concentration of 4 micrograms/ml inhibited only 31% of tested strains. For doxycicline 77% of strains were susceptible. Only five isolates were susceptible to clinafloxacin; the other quinolones tested displayed poor activity. Resistance to chloramphenicol was detected in 47% of isolates. None of the isolates produced beta-lactamase. This study indicates that vancomycin-resistant enterococci are often concomitantly resistant to multiple antibiotics. Quinupristin/dalfopristin was the most active agent tested against E. faecium strains. On the basis of these results quinupristin/dalfopristin could be a therapeutic option for the treatment of vancomycin-resistant E. faecium infections.


Evaluation of online education about diabetes management in the school setting.
There are a variety of initiatives to provide education to improve the quality of care for children with diabetes in the school setting. This study piloted and evaluated an online continuing education program for school nurses about diabetes management for children in schools using current practice principles. The evaluation determined if objectives of the program were met, if participants believed the education provided would enhance their ability to manage children with diabetes, and participants' opinions about the online delivery method. Nineteen participants were recruited from among school nurses unable to attend statewide face-to-face diabetes workshops. The majority reported that the objectives were met, and 91% reported the education would enhance their ability to manage students with diabetes. Whereas 33% preferred online continuing education, 17% preferred face-to-face education, and 50% were unsure. Because of the increasing prevalence of diabetes in children, it is important that school nurses have access to continuing education that provides current principles for diabetes management for children. The method of delivery should be determined by the learner's learning style preferences.


Use of Entero-Test, a simple approach for non-invasive clinical evaluation of the biliary disposition of drugs.
To evaluate the non-invasive collection of bile from healthy human subjects for the qualitative characterization of the biliary disposition of a drug, using spectrometric techniques. Twenty subjects underwent non-invasive bile capture using a peroral string test (Entero-Test) device prior to and following a single oral dose of simvastatin (80 mg). The device, consisting of a weighted gelatin capsule containing a highly absorbent nylon string, was swallowed by each subject with the proximal end of the string taped to the face. Once the weighted string was judged to have reached the duodenum, gallbladder contraction was stimulated in order to release bile. The string was then retrieved via the mouth, and bile samples were analysed for drug-related material using spectrometric and spectroscopic techniques following solvent extraction. Numerous metabolites of simvastatin were detected, and the major metabolites were consistent with those reported from studies where bile was collected using invasive techniques from patients dosed with [(14) C]-simvastatin. The results from this study demonstrate the utility of deploying the Entero-Test in human studies to provide structural information on biliary metabolites. This can be readily applied in drug development studies, including those in the target patient population and may eliminate the need for more invasive sampling techniques.


Outbreak of Crimean-Congo haemorrhagic fever in Quetta, Pakistan: contact tracing and risk assessment.
In December 1994 in a private hospital in Quetta, Pakistan, 3 health-workers contracted Crimean-Congo haemorrhagic fever (CCHF) after surgery on a bleeding patient who later died. We conducted a retrospective study to determine transmission risks among contacts. Fifty contacts gave blood for antibody tests and answered questions about exposure. Two of four people exposed percutaneously and one of five with cutaneous exposure contracted CCHE The person with cutaneous exposure was a surgeon who tore his glove during surgery and noted blood on his hand but no cut. There were no anti-CCHF antibodies or CCHF cases among persons whose skin came into contact with body fluids other than blood (0/4), who had skin-to-skin contact (0/16) with patients or were physically close to them (0/21). Three index case relatives reported that although 10 family members had cutaneous exposure, none developed CCHF. The family refused blood tests. CCHF transmission in resource-constrained settings can be limited by focusing on avoiding health worker contact with blood.


[Transcription of ribosomal protein genes rplKAJL and RNA-polymerase genes rpoBC in Escherichia coli cells: metabolic regulation of attenuation and the effect of rifampicin].
The E. coli genes rplKAJL specifying ribosomal proteins L11, L1, L10, L7/L12 are co-transcribed with the genes rpoBC encoding the beta- and beta'-subunits of RNA polymerase, but are separated by the site of attenuation. The efficiency of attenuation within rplKAJL-rpoBC operon was determined as a ratio of rplKAJL transcription frequency to the same of rpoBC genes. The efficiency of attenuation was found to be a growth-rate dependent parameter of E. coli cells. At growth rate 1.2 doublings per hour the attenuation is rare and simultaneously increases with the increase in the growth rate (at mu = 1.2 doublings per hour the efficiency of attenuation is 4). Rifampicin (10-30 micrograms/ml) inhibits the transcription of both rplKAJL and rpoBC genes in fast growing cells but paradoxically stimulates their transcription in slowly growing cells. The stimulatory effect of rifampicin on rplKAJL genes transcription is supposed to be based on its ability to repress the ppGpp synthesis. The possible role of ppGpp in the regulation of transcription attenuation in rplKAJL-rpoBC operon is discussed.


In situ synthesis of fluorescent polydopamine nanoparticles coupled with enzyme-controlled dissolution of MnO2 nanoflakes for a sensitive immunoassay of cancer biomarkers.
Herein, a novel immunosensing strategy was designed for the sensitive fluorescence/visual detection of alpha-fetoprotein (AFP), by combining the enzyme-controlled formation of polydopamine (PDA) nanoparticles with the dissolution of manganese dioxide (MnO2) nanoflakes. Initially, a sandwich-type immunoreaction was fabricated on a capture antibody-coated microplate by using detection antibody/ascorbate oxidase-conjugating gold nanoparticles (AOX-AuNP-Ab2) as the detection probe. The ascorbate oxidase, carried within the immunocomplex, oxidized ascorbic acid (AA) to dehydroascorbic acid (DAA) to inhibit the dissolution of the MnO2 nanoflakes. This resulted in an increment in fluorescent PDA nanoparticles from the oxidation of dopamine (DA) into its quinone derivative with the aid of the MnO2 nanoflakes. Under optimal conditions, the fluorescence intensity of the PDA nanoparticles increased with increasing AFP concentration within the dynamic working range of 0.05-20 ng mL-1, with a detection limit (LOD) of 17.3 pg mL-1. The sensor also showed high specificity and acceptable accuracy for the detection of target AFP against other potential cancer biomarkers. Importantly, this study demonstrated for the first time that fluorescent PDA nanoparticles could be used as signal-generation tags and applied to an enzyme immunoassay. Although this methodology focused on AFP detection, it would be suitable for the detection of other cancer biomarkers if the corresponding antibodies were employed.


Clinical Validation of a Predictive Model for the Presence of Cervical Lymph Node Metastasis in Papillary Thyroid Cancer.
Ultrasound is a standard technique to detect lymph node metastasis in papillary thyroid cancer. Cystic changes and microcalcifications are the most specific features of metastasis, but with low sensitivity. This prospective study compared the diagnostic accuracy of a predictive model for sonographic evaluation of lymph nodes relative to the radiologist's standard assessment in detecting papillary thyroid cancer metastasis in patients after thyroidectomy. Cervical lymph node sonographic images were reported by a radiologist (R method) per standard practice. The same images were independently evaluated by another radiologist using a sonographic predictive model (M method). A test was considered positive for metastasis if the R or M method suggested lymph node biopsy. The result of lymph node biopsy or surgical pathology was used as the reference standard. We estimated relative true-positive fraction and relative false-positive fraction using log-linear models for correlated binary data for the M method compared with the R method. A total of 237 lymph nodes in 103 patients were evaluated. Our analysis of relative true-positive fraction and relative false-positive fraction included 54 nodes with pathologic results in which at least 1 method (R or M) was positive. The M method had a higher relative true-positive fraction of 1.46 (95% CI, 1.12-1.91; P = .006) and a lower relative false-positive fraction of 0.58 (95% CI, 0.36-0.92; P = .02) compared with the R method. The sonographic predictive model outperformed the standard assessment to detect lymph node metastasis in patients with papillary thyroid cancer and may reduce unnecessary biopsies.


Effects of CO(2) enrichment on growth and root (15)NH(4) (+) uptake rate of loblolly pine and ponderosa pine seedlings.
We examined changes in root growth and (15)NH(4) (+) uptake capacity of loblolly pine (Pinus taeda L.) and ponderosa pine (Pinus ponderosa Douglas. Ex Laws.) seedlings that were grown in pots in a phytotron at CO(2) partial pressures of 35 or 70 Pa with NH(4) (+) as the sole N source. Kinetics of (15)N-labeled NH(4) (+) uptake were determined in excised roots, whereas total NH(4) (+) uptake and uptake rates were determined in intact root systems following a 48-h labeling of intact seedlings with (15)N. In both species, the elevated CO(2) treatment caused a significant downregulation of (15)NH(4) (+) uptake capacity in excised roots as a result of a severe inhibition of the maximum rate of root (15)NH(4) (+) uptake (V(max)). Rates of (15)NH(4) (+) uptake in intact roots were, however, unaffected by CO(2) treatment and were on average 4- to 10-fold less than the V(max) in excised roots, suggesting that (15)NH(4) (+) absorption from the soil was not limited by the kinetics of root (15)NH(4) (+) uptake. Despite the lack of a CO(2) effect on intact root absorption rates, (15)NH(4) (+) uptake on a per plant basis was enhanced at high CO(2) concentrations in both species, with the relative increase being markedly higher in ponderosa pine than in loblolly pine. High CO(2) concentration increased total (15)NH(4) (+) uptake and the fraction of total biomass allocated to fine roots (< 2 mm in diameter) to a similar relative extent. We suggest that the increased uptake on a per plant basis in response to CO(2) enrichment is largely the result of a compensatory increase in root absorbing surfaces.


Determining the prevalence and genetic diversity of plasmid-mediated sulfonamide resistance in Escherichia coli from commercial broiler samples.
Sulfonamides are commonly used antibacterials in commercial poultry, contributing toward the development of multidrug-resistant (MDR) phenotypes among Escherichia coli and that has emerged as global concern. The current study aimed to assess the sulfonamide resistance among isolated E. coli strains among commercial broilers. The bacterial strains were identified from fecal samples (n = 100) using selective media, followed by initial identification based on biochemical profiles. The susceptibility was determined by measuring the minimum inhibitory concentration (MIC) against sulfamethoxazole. The study also evaluated mobile genetic elements (MGEs), the mediators of antibiotic resistance, by amplification of plasmid DNA using specific primer PCR. Additionally, the isolates were subjected to multilocus sequence typing (MLST) analysis to investigate the genetic diversity among E. coli carrying sulfonamide resistance genes. The results revealed that 58% (58/100) E. coli strains were resistant to sulfonamides, with 36.20% (21/58) of the strains exhibiting an MIC breakpoint ≥512 µg/mL. PCR analysis showed that 42.85% (9/21) of the strains harbored the sul-1 gene, while 38.09% (8/21) carried the sul-2 gene, and 19.04% (4/21) had both genes. No isolate showed the presence of the sul-3 gene. Furthermore, class 1 and class 2 integrons were identified among 80.95% (17/21) and 19.04% (4/21) of the strains, respectively. MLST analysis confirmed that the strains belonged to sequence types (STs) including ST1638, ST155, ST48, ST350, ST23, ST156, and ST746. These findings underscore the diversity among E. coli strains in commercial poultry, which poses a significant risk.


Efficacy of annealing and fabrication parameters on photo-response of SiGe in TiO2 matrix.
SiGe nanoparticles dispersed in a dielectric matrix exhibit properties different from those of bulk and have shown great potential in devices for application in advanced optoelectronics. Annealing is a common fabrication step used to increase crystallinity and to form nanoparticles in such a system. A frequent downside of such annealing treatment is the formation of insulating SiO2 layer at the matrix/SiGe interface, degrading the optical properties of the structure. An annealing process that could bypass this downside would therefore be of great interest. In this work, a short-time furnace annealing of a SiGe/TiO2 system is applied to obtain SiGe nanoparticles without formation of the undesired SiO2 layer between the dielectric matrix (TiO2) and SiGe. The structures were prepared by depositing alternate layers of TiO2 and SiGe films, using direct-current magnetron sputtering technique. A wide range spectral response with a response-threshold up to ∼1300 nm was obtained, accompanied with an increase in photo-response of more than two-orders of magnitude. Scanning electron microscopy, transmission electron microscopy, energy-dispersive x-ray spectroscopy and grazing incidence x-ray diffraction were used to analyze the morphological changes in respective structures. Photoconductive properties were studied by measuring photocurrent spectra using applied dc-voltages at various temperatures.


Physiological changes in skeletal muscle as a result of strength training.
The picture of training that emerges is of a process that can be divided into a number of phases. In the first phase there is a rapid improvement in the ability to perform the training exercise such as lifting weights which is the result of a learning process in which the correct sequence of muscle contractions is laid down as a motor pattern in the central nervous system. This phase is associated with little or no increase in the size or strength of individual muscles. The learning process appears to be very specific in that lifting weights makes better weight lifters but not better sprinters. The second phase is an increase in the strength of individual muscles which occurs without a matching increase in the anatomical cross-section. The mechanism for this is not clear but could be a result of increased neural activation or some change in the fibre arrangement or connective tissue content. The third phase starts at a point where scientific studies usually end, at about 12 weeks when non-athletic subjects are beginning to tire of the repeated training and testing. After this point, if training continues, there is probably a slow but steady increase in both size and strength of the exercised muscles. The stimulus for these changes remains enigmatic but almost certainly involves high forces in the muscle, probably to induce some form of damage that promotes division of satellite cells and their incorporation into existing muscle fibres. Our information on the effect of long-term training comes primarily from observations on elite athletes whose physique may well be the result of genetic endowment or the use or abuse of drugs. For the athlete or patient hoping to increase muscle size by weight training the best combination of intensity, frequency and type of exercise still remains a matter of individual choice rather than a scientific certainty.


Drought and salt tolerance enhancement of transgenic Arabidopsis by overexpression of the vacuolar pyrophosphatase 1 (EVP1) gene from Eucalyptus globulus.
Vacuolar solute accumulation has been shown to be a mechanism by which plants are capable of increasing drought and salt tolerance. The exposure of plants to NaCl induces H+ transport into the vacuole by specialized pumps. One of them corresponds to the vacuolar H+-pyrophosphatase, which generates a H+ gradient across the vacuolar membrane. In our laboratory we isolated the first cDNA sequence of a vacuolar pyrophosphatase type I (EVP1) from Eucalyptus globulus. Using real-time PCR we confirmed that EVP1 participates in Eucalyptus plants' response to drought and salt stress through an ABA independent pathway. Additionally, the overexpression of EVP1 in transgenic Arabidopsis resulted in an enhancement of drought and salt tolerance. Interestingly we established that the transgenic plants had a higher number of root hairs, which may have a positive effect on the plant's response to drought and salt stress. These results suggest that EVP1 plays an active role in abiotic stress tolerance in E. globulus, and that it may be potentially used to enhance drought and stress tolerance of plants.


Subcellular localization and properties of partially purified dimethylamine and trimethylamine mono-oxygenase activities in Candida utilis.
By techniques involving differential centrifugation and specific precipitation with CaCl2, it was shown that dimethylamine and trimethylamine mono-oxygenase activities co-sediment with NADPH-cytochrome c reductase activity in sphaeroplast lysates of Candida utilis grown on trimethylamine as sole nitrogen source. Since the active fraction also contained low levels of cytochromes P-450 and P-420, it was concluded that the two amine mono-oxygenases are located in the smooth endoplasmic reticulum and thus end up in the microsomal fraction on cell fractionation. Ten to twenty-fold enrichment of mono-oxygenase specific activity could be achieved by separation of activity from soluble protein by centrifugation or gel filtration. Cell-free extracts prepared in the absence of FAD showed only very low mono-oxygenase activity for either substrate. Some activity could be restored by addition of flavin nucleotides: there was a fivefold stimulation by FAD and a fourfold stimulation by FMN. All trimethylamine mono-oxygenase activity was lost when a partially purified preparation containing both activities was incubated for more than 24 h at 0 degrees C, suggesting that separate enzymes are responsible for the oxidation of secondary and tertiary amines. The enzyme preparation oxidized a wide range of secondary alkylamines up to dibutylamine and tertiary alkylamines up to tributylamine. Primary amines, choline, di- and triethanolamine, spermine, spermidine and substituted anilines were not oxidized. NADH had a lower apparent Km value and higher Vmax value than NADPH. Secondary and tertiary alkylamines containing more than one kind of alkyl group gave more than one kind of aldehyde on oxidation. Stoicheiometry determinations showed a consumption of 1 mol NAD(P)H and 1 mol O2 per mol aldehyde formed. Carbon monoxide, cyanide, proadifen hydrochloride (SKF 525-A), mercurials and mercaptoethanol all inhibited both activities.


[Automatic generation of gene network schemes based on their formalized description in the GeneNet database].
A database for gene networks GeneNet was created. The main principles of formalized description of gene networks for the cases of regulation of antiviral responses and hemopoiesis were developed. A program for automatic graphical representation of diagrams of gene networks was created, which is based on their formalized description. A system of filters was developed, which makes it possible to select individual network components for graphical representation. The selection can be performed with respect to specific type of treatment, species, and/or cell type. The GeneNet database is accessible via Internet at http://wwwmgs.bionet.nsc.ru/systems/MGL/Gen eNet.


Interprofessional global health education in a cosmopolitan community of North America: the iCHEE experience.
The rapidly diversifying population of North America has disparate health needs that are addressed by creative, community-based training of health professions students. The authors report five years (2008-2012) of experience implementing a novel interprofessional Community Health and Education Exchange (iCHEE) elective course for dental, medical, nursing, nutrition, pharmacy, physician assistant, and public health students at Oregon Health & Science University (OHSU). This pioneering interprofessional course was created by the OHSU Global Health Center and is offered in fall, winter, and spring quarters. Students interact with individual clients drawn from community centers supporting refugees, recent immigrants, and other underserved people. In addition to health concerns, clients are encouraged to share backgrounds and experiences with student teams. Clients receive guidance on nutrition, exercise, pharmaceuticals, and accessible health services. Student teams perform a noninvasive health check on clients with the assistance of faculty mentors who, on finding a physical or mental health issue, refer the client from the educational setting to an appropriate health care facility. In addition to supporting health promotion and early intervention for medically underserved people, students reported gaining valuable cross-cultural knowledge, understanding, and experience from clients. Students also appreciated the value of diverse skills and knowledge available in their multidisciplinary teams. Through the end of 2012, over 300 health professions students worked with approximately 1,200 clients to complete the iCHEE course. The iCHEE model should prove helpful in preparing health professions students at other institutions to understand and serve diverse populations.


Potential inhibition of the neuro-neoplastic interactions: the clue of a GPCR-targeted therapy.
Other sections of this monograph, dedicated to neuronal activities in tumor tissue, have highlight the chief influence of neurotrophins, neurotransmitters, adhesion, guidance molecules and different nerve cell markers in the progression, but also for the prognostic, therapy and survey of cancers. The G-protein-coupled receptors (GPCR) are among the most successful and promising target proteins for drug discovery and therapeutic research. GPCR are frequently overexpressed in cancer cells, an interesting property for tumor imaging or for a targeted radiotherapy, using radiolabeled ligand derivatives. The tumor microenvironment contains a number of GPCR ligands (e.g., bioactive peptides, biogenic amines, purins, chemokines), known to regulate the proliferation, migration or survival of both tumoral and neural cells and that may be key actors of the neuro-neoplastic interactions. Here will be reviewed the potential utilization of substances that target a selected choice of GPCR, especially neuropeptide receptors, for a novel concept of therapy, concerning the numerous types of cancers where neurons infiltrate the tumoral mass or those where the malignant cells invade nerve branches (perineural invasion). Some molecular mechanisms linked to these GPCR (or linking GPCR to other types of membrane receptors or co-receptors), involved in these processes, will also be considered.


Genetic polymorphisms in key methotrexate pathway genes are associated with response to treatment in rheumatoid arthritis patients.
We investigated the effect of single-nucleotide polymorphisms (SNPs) spanning 10 methotrexate (MTX) pathway genes, namely AMPD1, ATIC, DHFR, FPGS, GGH, ITPA, MTHFD1, SHMT1, SLC19A1 (RFC) and TYMS on the outcome of MTX treatment in a UK rheumatoid arthritis (RA) patient cohort. Tagging SNPs were selected and genotyping was performed in 309 patients with predefined outcomes to MTX treatment. Of the 129 SNPs tested, 11 associations were detected with efficacy (P-trend 0.05) including four SNPs in the ATIC gene (rs12995526, rs3821353, rs7563206 and rs16853834), six SNPs in the SLC19A1 gene region (rs11702425, rs2838956, rs7499, rs2274808, rs9977268 and rs7279445) and a single SNP within the GGH gene (rs12681874). Five SNPs were significantly associated with adverse events; three in the DHFR gene (rs12517451, rs10072026, and rs1643657) and two of borderline significance in the FPGS gene. The results suggest that genetic variations in several key MTX pathway genes may influence response to MTX in the RA patients. Further studies will be required to validate these findings and if confirmed these results could contribute towards a better understanding of and ability to predict MTX response in RA.


Network analysis reveals multiscale controls on streamwater chemistry.
By coupling synoptic data from a basin-wide assessment of streamwater chemistry with network-based geostatistical analysis, we show that spatial processes differentially affect biogeochemical condition and pattern across a headwater stream network. We analyzed a high-resolution dataset consisting of 664 water samples collected every 100 m throughout 32 tributaries in an entire fifth-order stream network. These samples were analyzed for an exhaustive suite of chemical constituents. The fine grain and broad extent of this study design allowed us to quantify spatial patterns over a range of scales by using empirical semivariograms that explicitly incorporated network topology. Here, we show that spatial structure, as determined by the characteristic shape of the semivariograms, differed both among chemical constituents and by spatial relationship (flow-connected, flow-unconnected, or Euclidean). Spatial structure was apparent at either a single scale or at multiple nested scales, suggesting separate processes operating simultaneously within the stream network and surrounding terrestrial landscape. Expected patterns of spatial dependence for flow-connected relationships (e.g., increasing homogeneity with downstream distance) occurred for some chemical constituents (e.g., dissolved organic carbon, sulfate, and aluminum) but not for others (e.g., nitrate, sodium). By comparing semivariograms for the different chemical constituents and spatial relationships, we were able to separate effects on streamwater chemistry of (i) fine-scale versus broad-scale processes and (ii) in-stream processes versus landscape controls. These findings provide insight on the hierarchical scaling of local, longitudinal, and landscape processes that drive biogeochemical patterns in stream networks.


High-Pitch Wide-Coverage Fast-Kilovoltage-Switching Dual-Energy CT: Impact of Pitch on Noise, Spatial Resolution, and Iodine Quantification in a Phantom Study.
The purpose of this study was to assess the impact of high pitch values on image noise, spatial resolution, and iodine quantification in single-source wide-coverage fast-kilovoltage-switching dual-energy CT (DECT). Two phantom experiments were conducted. First, image noise and spatial resolution in the x-, y-, and z-directions were assessed. Second, the accuracy of iodine quantification was investigated with multiple-size phantoms with pure iodine and blood-iodine inserts. Both phantoms were scanned repeatedly with a third-generation fast-kilovoltage-switching DECT scanner with a collimation width of 80 mm at four different pitch values (0.5, 0.99, 1.375, 1.53) and three different gantry rotation times (0.6, 0.8, 1.0 second). Image noise, spatial resolution, and absolute error of iodine concentration (E) were measured. A linear mixed effects model was used to determine the effect of pitch, rotation time, and size on the error of iodine concentration. Image noise and xy spatial resolution were comparable among the four pitch values. Spatial resolution in the z-direction was inferior and had higher variance at a low pitch of 0.5 compared with pitches of 0.99, 1.375, and 1.53. Error of iodine concentration was significantly affected by pitch and rotation time (p < 0.001). E decreased with increasing pitch and decreasing rotation time. In detail, mean E was 0.91 ± 0.47 mg I/mL for a pitch of 0.5, 0.52 ± 0.29 mg I/mL for 0.99, 0.44 ± 0.25 mg I/mL for 1.375, and 0.40 ± 0.25 mg I/mL for 1.53. High-pitch wide-coverage fast-kilovoltage-switching DECT can be performed without impairing image quality or iodine quantification, and the results are superior to those of imaging at a low pitch of 0.5.


Risk factors, mortality, and predictors of survival in COVID-19-associated pulmonary mucormycosis: a multicenter retrospective study from India.
To compare COVID-19-associated pulmonary mucormycosis (CAPM) with COVID-19-associated rhino-orbital mucormycosis (CAROM), ascertain factors associated with CAPM among patients with COVID-19, and identify factors associated with 12-week mortality in CAPM. We performed a retrospective multicenter cohort study. All study participants had COVID-19. We enrolled CAPM, CAROM, and COVID-19 subjects without mucormycosis (controls; age-matched). We collected information on demography, predisposing factors, and details of COVID-19 illness. Univariable analysis was used to compare CAPM and CAROM. We used multivariable logistic regression to evaluate factors associated with CAPM (with hypoxemia during COVID-19 as the primary exposure) and 12-week mortality. We included 1,724 cases (CAPM [n=122], CAROM [n=1,602]) and 3,911 controls. Male sex, renal transplantation, multimorbidity, higher neutrophil-lymphocyte ratio, intensive care admission, and cumulative glucocorticoid dose for COVID-19 were significantly higher in CAPM than in CAROM. On multivariable analysis, COVID-19-related hypoxemia (adjusted odds ratio [aOR] 2.388, 95% confidence interval [CI] 1.209-4.700), male sex, rural residence, diabetes, serum C-reactive protein, glucocorticoid and zinc use during COVID-19 were independently associated with CAPM. CAPM had a higher 12-week mortality than CAROM (56/107 [52.3%] vs. 413/1356 [30.5%]; p=0.0001). Hypoxemia during COVID-19 (aOR [95% CI], 3.70 [1.34-10.25]) and Aspergillus co-infection (aOR [95% CI], 5.40 [1.23-23.64]) were independently associated with mortality in CAPM, while surgery was associated with better survival. CAPM is a distinct entity with a higher mortality than CAROM. Hypoxemia during COVID-19 illness is associated with CAPM. COVID-19 hypoxemia and Aspergillus co-infection were associated with higher mortality in CAPM.


Epidemiological and genetic characteristics of asymptomatic canine leishmaniasis and implications for human Leishmania infections in Egypt.
Leishmaniasis is a neglected zoonotic disease that poses significant veterinary and public health risks in developing countries. Dogs act as a reservoir host for leishmaniasis transmitted to humans. A total of 108 human cases of cutaneous leishmaniasis (CL) were identified in the Al-Houd Al-Marsoud Hospital in Cairo, Egypt, during 2018. Blood samples and skin biopsies were collected for further examination. Blood samples from 96 asymptomatic dogs were collected. All samples were subjected to molecular and phylogenetic analysis. Quantitative RT-PCR was used to measure the expression of genes related to mTOR signalling and inflammation in blood and tissue samples. The distribution pattern of human cases pointed to an endemic focus in North Sinai (66.67%). The prevalence of asymptomatic canine leishmaniasis was 66.60%. Histopathological examination of human skin lesions revealed a severe granulomatous inflammatory reaction, necrosis and ulceration. Moreover, leishmanial amastigotes could be detected in human tissue samples. Phylogenetic analysis revealed 100% identity of human isolates to Leishmania tropica (MN453682), and dog isolates to Leishmania infantum (MN453673), with 94.9% similarity between the two isolates. Gene expression related to mTOR signalling and inflammation in both species' samples confirmed a significant alteration of EIF4EBP1, CCR4 and INF-γ expression compared with control groups. In Egypt, increased incidence of asymptomatic carrier dogs acting as a significant reservoir host for Leishmania poses a public health hazard. Findings warrant further epidemiological investigation of CL in Egypt, as well as additional study of parasite differentiation and gene regulation.


Efficient Generation of Chondrocytes From Bone Marrow-Derived Mesenchymal Stem Cells in a 3D Culture System: Protocol for a Practical Model for Assessing Anti-Inflammatory Therapies.
Chondrocytes are the primary cells responsible for maintaining cartilage integrity and function. Their role in cartilage homeostasis and response to inflammation is crucial for understanding the progression and potential therapeutic interventions for various cartilage-related disorders. Developing an accessible and cost-effective model to generate viable chondrocytes and to assess their response to different bioactive compounds can significantly advance our knowledge of cartilage biology and contribute to the discovery of novel therapeutic approaches. We developed a novel, streamlined protocol for generating chondrocytes from bone marrow-derived mesenchymal stem cells (BMSCs) in a 3D culture system that offers significant implications for the study of cartilage biology and the discovery of potential therapeutic interventions for cartilage-related and associated disorders. We developed a streamlined protocol for generating chondrocytes from BMSCs in a 3D culture system using an "in-tube" culture approach. This simple pellet-based 3D culture system allows for cell aggregation and spheroid formation, facilitating cell-cell and cell-extracellular matrix interactions that better mimic the in vivo cellular environment compared with 2D monolayer cultures. A proinflammatory chondrocyte model was created by treating the chondrocytes with lipopolysaccharide and was subsequently used to evaluate the anti-inflammatory effects of vitamin D, curcumin, and resveratrol. The established protocol successfully generated a large quantity of viable chondrocytes, characterized by alcian blue and toluidine blue staining, and demonstrated versatility in assessing the anti-inflammatory effects of various bioactive compounds. The chondrocytes exhibited reduced inflammation, as evidenced by the decreased tumor necrosis factor-α levels, in response to vitamin D, curcumin, and resveratrol treatment. Our novel protocol offers an accessible and cost-effective approach for generating chondrocytes from BMSCs and for evaluating potential therapeutic leads in the context of inflammatory chondrocyte-related diseases. Although our approach has several advantages, further investigation is required to address its limitations, such as the potential differences between chondrocytes generated using our protocol and those derived from other established methods, and to refine the model for broader applicability and clinical translation.


Kinetics of bicarbonate transport in the early proximal convoluted tubule.
Bicarbonate permeability and bicarbonate transport kinetics in the S1 segment of the proximal convoluted tubule (PCT) have not been previously studied. In vivo microperfusion at a rate of 30 nl/min was performed in early and late PCT of Munich-Wistar rats. Bicarbonate permeability was first defined, using a bicarbonate-free, acetazolamide-containing perfusate, and was over fourfold higher in the early compared with the late PCT (20.4 +/- 1.8 vs. 4.6 +/- 0.4 X 10(-7) cm2/s, P less than 0.001). Net bicarbonate absorption was then measured using perfusate bicarbonate concentrations of 15, 25, 40, and 100 mM at 30 nl/min perfusion rate. Proton secretory rate was calculated for each group by subtracting the passive bicarbonate transport component from the net flux. Saturation kinetics of acidification were observed in both the early and late PCT. The maximal proton secretory rate at the highest luminal bicarbonate concentration (Vmax) in the early PCT was about twice that in the late PCT (504 +/- 37 vs. 265 +/- 15 peq.mm-1.min-1, P less than 0.001). However, the luminal bicarbonate concentration eliciting half-maximal proton secretion (apparent Km) was approximately the same (11 mM) in the early and late PCT. In conclusion, the early PCT has a higher bicarbonate permeability and proton secretory capacity than the late PCT. Increased Vmax but axial constancy of Km suggests that there is amplification of similar transport mechanism(s) affecting bicarbonate absorption in S1 compared with S2 cells.


[Complications of thyroid surgery. Report of 683 thyroidectomies].
Most important complications of thyroid gland surgery are recurrent paralysis (which usually results from section, stretching or devascularization of this neural branch) and parathyroid lesions resulting from unnoticed removal of more than two of these glands or through necrosis because deficient blood supply. We present a study about the complications of thyroid surgery based on the thyroid surgery done in our ENT Service during 1979-1998 term. An amount of 683 cases make the series. The 86% were female and 14% male, with a mean age of 40-50. Surgical techniques realized were: hemithyroidectomy (15.3%) and isthmectomy (2.9%). About 7.4% of patients were reoperated. Stated complications were: local (serohematoma 4.1%, infection 1.2%, postoperative hemorrhage 1.2%) and recurrent paralysis (unilateral transitory 2.6% and unilateral definitive 1.6%; there wasn't any case of bilateral palsy) and superior laryngeal nerve paralysis (1.4% as well as hypoparathyroidism (transitory 22.7% and definitive 5.3%). These complications were most frequent in reoperated cases and in total and subtotal thyroidectomies. In most of these series the incidence of recurrent palsy range between 1.5% and increases more than 9.5% in surgical check and the hypoparathyroidism incidence ranges over 25%.


Tolerance induced by bone marrow chimerism prevents transplant vascular sclerosis in a rat model of small bowel transplant chronic rejection.
The major impediment to success in solid organ transplantation is chronic rejection (CR). The characteristic lesion of CR is transplant vascular sclerosis (TVS). Although the mechanism of TVS is thought to have an immunologic basis, in humans immunosuppression does not prevent or reverse it. One possible therapy to prevent TVS is induction of donor-specific tolerance. Bone marrow chimerism has been successful in inducing tolerance in acute and chronic rejection heart and kidney transplant models. The highly immunogenic small bowel (SB) allograft provides a rigorous test of the efficacy of this tolerance regimen. We examined whether induction of tolerance by bone marrow chimerism could prevent TVS in a model of Fisher 344 (F344) to Lewis (LEW) rat SB transplantation. Bone marrow chimeras (BMC) were created by transplantation of T-cell-depleted F344 bone marrow into irradiated LEW rats. Chimerism was assessed by flow cytometric method. F344 SB, heterotopically transplanted into the chimeras, was clinically and histologically assessed for CR. F344 SB grafts, transplanted into cyclosporine-A-treated LEW recipients, served as control grafts for CR. Cyclosporine-A-treated LEW rats chronically rejected F344 SB grafts. By contrast, the BMC group demonstrated tolerance and had long-term SB graft survival (>120 days) without TVS. The BMC demonstrated immunocompetence by prompt rejection of third party ACI (RT1av1) SB allografts. Bone marrow chimerism prevents chronic graft failure secondary to TVS in a model of chronic SB rejection. TVS fails to develop when tolerance is established, suggesting that the mechanisms involved in TVS are, in part, immunologically mediated.


Stigma Mechanisms and Outcomes among Sub-Saharan African Descendants in Belgium-Contextualizing the HIV Stigma Framework.
HIV-related stigma and discrimination are recognized barriers to HIV prevention, testing and treatment among people of Sub-Saharan African descent (SSA) origin living in Belgium, but insights into HIV related-stigma mechanisms and outcomes are lacking for this population with high HIV prevalence. Guided by Earnshaw and Chaudoir's stigma framework (2009), we conducted this qualitative study using 10 focus-groups with 76 SSA community members and 20 in-depth interviews with SSA descendants living with HIV to explore specific HIV-stigma mechanisms and outcomes and underlying drivers. Inductive and deductive thematic analysis showed high degrees of stigma among SSA communities driven by fear of HIV acquisition and misconceptions in a migration context, negatively affecting SSA descendants living with HIV. The results allowed for contextualization of the framework: At the community level, prejudices and stereotypes were major stigma mechanisms, while physical distancing, gossips, sexual rejection, violence and increased HIV prevalence emerged as stigma outcomes. Among SSA descendants living with HIV, enacted, anticipated and internalized stigmas were validated as stigma mechanisms, with witnessed stigma as an additional mechanism. Self-isolation, community avoidance and low utilization of non-HIV specialized healthcare were additional outcomes. These results are relevant for tailoring interventions to reduce HIV-related stigma.


Using acupressure to modify alertness in the classroom: a single-blinded, randomized, cross-over trial.
Previous reports have suggested that acupressure is effective in reducing pain and improving sleep quality; however, its effects on alertness have not been characterized. The aim of this study was to determine whether two different acupressure treatments have opposing effects on alertness in a full-day classroom setting. This was a cross-over (two-treatments; three periods), single-blinded, randomized trial. The University of Michigan School of Public Health was the setting. Students attending a course in clinical research design and statistical analysis at the University of Michigan participated in the study. Blinded subjects were randomized to two acupressure treatment sequences: stimulation-relaxation-relaxation or relaxation-stimulation-stimulation. Acupressure treatments were self administered over 3 consecutive days. Pre- and post-treatment alertness scores were assessed each day using the Stanford Sleepiness Scale (SSS). Changes in the SSS score (afternoon-morning) were analyzed using a mixed regression model of fixed and random effects. Important factors that were expected to affect alertness, such as caffeine and previous night's sleep, were also assessed. Baseline characteristics and protocol compliance were similar between the two sequences. Stimulation acupressure treatment yielded a 0.56-point greater difference in score on the SSS, corresponding to less fatigue, compared to the relaxation acupressure treatment (p = 0.019). Day of study (p = 0.004) and hours of overnight sleep (p = 0.042) also significantly affected the change in SSS scores. Incorporating participants' beliefs as to which treatment they received did not significantly alter the observed treatment effect. Acupressure at stimulation and relaxation points has differential effects on alertness in a classroom setting. Further research is necessary to confirm these findings and to determine whether stimulation and relaxation acupressure are equally effective in influencing alertness.


Thermal comfort sustained by cold protective clothing in Arctic open-pit mining-a thermal manikin and questionnaire study.
Workers in the Arctic open-pit mines are exposed to harsh weather conditions. Employers are required to provide protective clothing for workers. This can be the outer layer, but sometimes also inner or middle layers are provided. This study aimed to determine how Arctic open-pit miners protect themselves against cold and the sufficiency, and the selection criteria of the garments. Workers' cold experiences and the clothing in four Arctic open-pit mines in Finland, Sweden, Norway and Russia were evaluated by a questionnaire (n=1,323). Basic thermal insulation (Icl) of the reported clothing was estimated (ISO 9920). The Icl of clothing from the mines were also measured by thermal manikin (standing/walking) in 0.3 and 4.0 m/s wind. The questionnaire showed that the Icl of the selected clothing was on average 1.2 and 1.5 clo in mild (-5 to +5°C) and dry cold (-20 to -10°C) conditions, respectively. The Icl of the clothing measured by thermal manikin was 1.9-2.3 clo. The results show that the Arctic open-pit miners' selected their clothing based on occupational (time outdoors), environmental (temperature, wind, moisture) and individual factors (cold sensitivity, general health). However, the selected clothing was not sufficient to prevent cooling completely at ambient temperatures below -10°C.


The large non-coding RNA ANRIL, which is associated with atherosclerosis, periodontitis and several forms of cancer, regulates ADIPOR1, VAMP3 and C11ORF10.
The long non-coding RNA ANRIL is the best replicated genetic risk locus of coronary artery disease (CAD) and periodontitis (PD), and is independently associated with a variety of other immune-mediated and metabolic disorders and several forms of cancer. Recent studies showed a correlation of decreased concentrations of proximal ANRIL transcripts with homozygous carriership of the CAD and PD main risk alleles. To elucidate the relation of these transcripts to disease manifestation, we constructed a short hairpin RNA in a stable inducible knock-down system of T-Rex 293 HEK cell lines, specifically targeting the proximal transcripts EU741058 and DQ485454. By genome-wide expression profiling using Affymetrix HG1.0 ST Arrays, we identified the transcription of ADIPOR1, VAMP3 and C11ORF10 to be correlated with decreased ANRIL expression in a time-dependent manner. We validated these findings on a transcriptional and translational level in different cell types. Exploration of the identified genes for the presence of disease associated variants, using Affymetrix 500K genotyping and Illumina custom genotyping arrays, highlighted a region upstream of VAMP3 within CAMTA1 to be associated with increased risk of CAD [rs10864294 P = 0.015, odds ratio (OR) = 1.30, 95% confidence interval (CI) = 1.1-1.6, 1471 cases, 2737 controls] and aggressive PD (AgP; P = 0.008, OR = 1.31, 95% CI = 1.1-1.6, 864 cases, 3664 controls). In silico replication in a meta-analysis of 14 genome-wide association studies of CAD of the CARDIoGRAM Consortium identified rs2301462, located on the same haplotype block, as associated with P = 0.001 upon adjustment for sex and age. Our results give evidence that specific isoforms of ANRIL regulate key genes of glucose and fatty acid metabolism.


Geographical origin of durum wheat studied by 1H-NMR profiling.
(1)H-Nuclear magnetic resonance (NMR) spectroscopy was used to detect metabolic profiles of wheat flour samples of different geographical and botanical origin. The NMR profiles were analyzed by multivariate statistical techniques in order to establish the origin of the samples. A linear model, able to discriminate among three different locations, was built achieving a prediction level of about 80% of correctly assigned samples. The principal classes of compounds responsible for the geographic origin discrimination were individuated in aromatic compounds and amino acids. The statistical modeling also indicated that botanical origin information is very poor in the NMR profiles of the analyzed wheat samples.


The F-BAR Cdc15 promotes contractile ring formation through the direct recruitment of the formin Cdc12.
In Schizosaccharomyces pombe, cytokinesis requires the assembly and constriction of an actomyosin-based contractile ring (CR). Nucleation of F-actin for the CR requires a single formin, Cdc12, that localizes to the cell middle at mitotic onset. Although genetic requirements for formin Cdc12 recruitment have been determined, the molecular mechanisms dictating its targeting to the medial cortex during cytokinesis are unknown. In this paper, we define a short motif within the N terminus of Cdc12 that binds directly to the F-BAR domain of the scaffolding protein Cdc15. Mutations preventing the Cdc12-Cdc15 interaction resulted in reduced Cdc12, F-actin, and actin-binding proteins at the CR, which in turn led to a delay in CR formation and sensitivity to other perturbations of CR assembly. We conclude that Cdc15 contributes to CR formation and cytokinesis via formin Cdc12 recruitment, defining a novel cytokinetic function for an F-BAR domain.


Prevalence and Correlates of Sexual Revictimization in Middle and High School Youth.
Research consistently shows that individuals who experience a sexual assault are at an increased risk to experience a subsequent sexual assault, a phenomenon known as sexual revictimization. However, little research has used a longitudinal methodology to examine rates and correlates of peer-to-peer sexual revictimization in middle and high school youth. The purpose of this study was to address this gap in the literature. Participants were 1,706 youth in Grades 7 to 10 (51.3% female) who completed surveys at the beginning and end of an academic year, approximately 6 months apart. Results showed that the vast majority (n = 1561, 90.4%) of youth reported no sexual victimization at either time point, 125 youth (7.2%) reported sexual victimization at one time point, and 40 youth (2.3%) reported sexual victimization at two time points. Among victims (n = 165), about one quarter (n = 40, 24.2%) were victimized at both time points. Binge drinking, depression, and suicidal thoughts were all higher among multiple and singular victims compared with nonvictims and among multiple victims compared with singular victims. Girls and sexual minorities were more likely to be multiple and singular victims compared with nonvictims and multiple victims compared with singular victims. Older youth were more likely to be multiple and singular victims compared with nonvictims. These data underscore the need for programming that concurrently focuses on mental health and sexual assault risk reduction, especially among high-risk youth.


Incidence of bicuspid valve related aortic dissection: a systematic review and meta-analysis.
The true incidence of bicuspid valve-related aortic dissection (AD) is extremely difficult to ascertain. This review aimed to provide the reported cumulative incidence of bicuspid aortic valve (BAV)-related AD in actively monitored study populations. Four electronic databases were used to perform literature searches. A meta-analysis of proportions or means were performed for categorical and continuous variables, as appropriate. Survival data was calculated from the aggregation of Kaplan-Meier (KM) curves from the included studies, where reported. A total of 4,330 patients were identified in eleven studies. A cumulative incidence of bicuspid valve-related AD of 0.6% across a median follow-up time of 9 years was identified. Actuarial survival across this monitored population at 1, 3, 5 and 10 years was 97.2%, 96.7%, 92.45%, and 81.1%, respectively. This systematic review and meta-analysis identified a low incidence of AD across the examined follow-up period. Large, prospective studies involving early identification of bicuspid valve pathology, recruitment, and follow-up of BAV cohorts with comparison to the baseline population are required to most accurately determine the outcomes of these patients.


Using time-varying models to estimate post-transplant survival in pediatric liver transplant recipients.
To distinguish clinical factors that have time-varying (as opposed to constant) impact upon patient and graft survival among pediatric liver transplant recipients. Using national data from 2002 through 2013, we examined potential clinical and demographic covariates using Gray's piecewise constant time-varying coefficients (TVC) models. For both patient and graft survival, we estimated univariable and multivariable Gray's TVC, retaining significant covariates based on backward selection. We then estimated the same specification using traditional Cox proportional hazards (PH) models and compared our findings. For patient survival, covariates included recipient diagnosis, age, race/ethnicity, ventilator support, encephalopathy, creatinine levels, use of living donor, and donor age. Only the effects of recipient diagnosis and donor age were constant; effects of other covariates varied over time. We retained identical covariates in the graft survival model but found several differences in their impact. The flexibility afforded by Gray's TVC estimation methods identify several covariates that do not satisfy constant proportionality assumptions of the Cox PH model. Incorporating better survival estimates is critical for improving risk prediction tools used by the transplant community to inform organ allocation decisions.


Unstable chromosome aberrations do not accumulate in normal human fibroblast after fractionated x-irradiation.
We determined the frequencies of dicentric chromosomes per cell in non-dividing confluent normal human fibroblasts (MRC-5) irradiated with a single 1 Gy dose or a fractionated 1 Gy dose (10X0.1 Gy, 5X0.2 Gy, and 2X0.5 Gy). The interval between fractions was between 1 min to 1440 min. After the completion of X-irradiation, the cells were incubated for 24 hours before re-plating at a low density. Then, demecolcine was administrated at 6 hours, and the first mitotic cells were collected for 42 hours. Our study demonstrated that frequencies of dicentric chromosomes in cells irradiated with a 1 Gy dose at different fractions were significantly reduced if the fraction interval was increased from 1 min to 5 min (p<0.05, χ2-test). Further increasing the fraction interval from 5 up to 1440 min did not significantly affect the frequency of dicentric chromosomes. Since misrejoining of two independent chromosome breaks introduced in close proximity gives rise to dicentric chromosome, our results indicated that such circumstances might be quite infrequent in cells exposed to fractionated X-irradiation with prolonged fraction intervals. Our findings should contribute to improve current estimation of cancer risk from chronic low-dose-rate exposure, or intermittent exposure of low-dose radiation by medical exposure.


The melanoma-associated antigen MAGE-D2 suppresses TRAIL receptor 2 and protects against TRAIL-induced apoptosis in human melanoma cells.
Emerging evidence has pointed to biological roles of melanoma-associated antigens (MAGEs) in cancer development, progression and resistance to treatment. However, the mechanisms involved remain to be fully elucidated. In this report, we show that one of the MAGE proteins, MAGE-D2, suppresses the expression of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) death receptor 2 (TRAIL-R2) and plays an important role in protecting melanoma cells from apoptosis induced by TRAIL. MAGE-D2 was commonly expressed at increased levels in melanoma cells compared with melanocytes. Although its inhibition by small interfering RNA (siRNA) did not cause cell death, it rendered melanoma cells more sensitive to TRAIL-induced apoptosis. This was associated with enhanced formation of TRAIL death-inducing signaling complex and up-regulation of TRAIL-R2, and was blocked by a recombinant TRAIL-R2/Fc chimeric protein or siRNA knockdown of TRAIL-R2. Regulation of TRAIL-R2 by MAGE-D2 appeared to be mediated by p53, in that knockdown MAGE-D2 did not up-regulate TRAIL-R2 in p53-null or mutant p53 melanoma cells. In addition, inhibition of MAGE-D2 did not result in up-regulation of TRAIL-R2 in wild-type p53 cell lines with p53 inhibited by short hairpin RNA. Indeed, knockdown of MAGE-D2 led to up-regulation of p53 due to a transcriptional increase. The regulatory effect of MAGE-D2 on TRAIL-R2 expression and TRAIL-induced apoptosis was recapitulated in studies on fresh melanoma isolates. Taken together, these results identify the expression of MAGE-D2 as an important mechanism that inhibit TRAIL-induced apoptosis and suggest that targeting MAGE-D2 may be a useful strategy in improving the therapeutic efficacy of TRAIL in melanoma.


The PINK1/Parkin pathway regulates mitochondrial dynamics and function in mammalian hippocampal and dopaminergic neurons.
PTEN-induced putative kinase 1 (PINK1) and Parkin act in a common pathway to regulate mitochondrial dynamics, the involvement of which in the pathogenesis of Parkinson's disease (PD) is increasingly being appreciated. However, how the PINK1/Parkin pathway influences mitochondrial function is not well understood, and the exact role of this pathway in controlling mitochondrial dynamics remains controversial. Here we used mammalian primary neurons to examine the function of the PINK1/Parkin pathway in regulating mitochondrial dynamics and function. In rat hippocampal neurons, PINK1 or Parkin overexpression resulted in increased mitochondrial number, smaller mitochondrial size and reduced mitochondrial occupancy of neuronal processes, suggesting that the balance of mitochondrial fission/fusion dynamics is tipped toward more fission. Conversely, inactivation of PINK1 resulted in elongated mitochondria, indicating that the balance of mitochondrial fission/fusion dynamics is tipped toward more fusion. Furthermore, overexpression of the fission protein Drp1 (dynamin-related protein 1) or knocking down of the fusion protein OPA1 (optical atrophy 1) suppressed PINK1 RNAi-induced mitochondrial morphological defect, and overexpression of PINK1 or Parkin suppressed the elongated mitochondria phenotype caused by Drp1 RNAi. Functionally, PINK1 knockdown and overexpression had opposite effects on dendritic spine formation and neuronal vulnerability to excitotoxicity. Finally, we found that PINK1/Parkin similarly influenced mitochondrial dynamics in rat midbrain dopaminergic neurons. These results, together with previous findings in Drosophila dopaminergic neurons, indicate that the PINK1/Parkin pathway plays conserved roles in regulating neuronal mitochondrial dynamics and function.


Galectin-1 dimers can scaffold Raf-effectors to increase H-ras nanoclustering.
Galectin-1 (Gal-1) dimers crosslink carbohydrates on cell surface receptors. Carbohydrate-derived inhibitors have been developed for cancer treatment. Intracellularly, Gal-1 was suggested to interact with the farnesylated C-terminus of Ras thus specifically stabilizing GTP-H-ras nanoscale signalling hubs in the membrane, termed nanoclusters. The latter activity may present an alternative mechanism for how overexpressed Gal-1 stimulates tumourigenesis. Here we revise the current model for the interaction of Gal-1 with H-ras. We show that it indirectly forms a complex with GTP-H-ras via a high-affinity interaction with the Ras binding domain (RBD) of Ras effectors. A computationally generated model of the Gal-1/C-Raf-RBD complex is validated by mutational analysis. Both cellular FRET as well as proximity ligation assay experiments confirm interaction of Gal-1 with Raf proteins in mammalian cells. Consistently, interference with H-rasG12V-effector interactions basically abolishes H-ras nanoclustering. In addition, an intact dimer interface of Gal-1 is required for it to positively regulate H-rasG12V nanoclustering, but negatively K-rasG12V nanoclustering. Our findings suggest stacked dimers of H-ras, Raf and Gal-1 as building blocks of GTP-H-ras-nanocluster at high Gal-1 levels. Based on our results the Gal-1/effector interface represents a potential drug target site in diseases with aberrant Ras signalling.


Opiates and medial hypothalamic knife cuts cause hyperphagia through different mechanisms.
Several experiments were performed to determine whether the hyperphagia caused by medial hypothalamic knife cuts and that induced by opiate agonists are mediated by a common mechanism. In the first set of experiments, male Sprague-Dawley rats were given bilateral parasagittal medial hypothalamic knife cuts or a sham procedure and fed a high-fat Crisco-chow diet. Knife-cut and sham-operated rats were approximately equally sensitive to the suppressive effects of naloxone on food intake. The kappa opiate receptor agonist ketocyclazocine generally increased daytime food intake in sham-operated rats; in contrast, the normal hyperphagia of knife-cut rats was in most cases either unchanged or decreased by ketocyclazocine. In a second set of experiments, neither diet composition nor hypothalamic knife cuts significantly affected the feeding responses to naloxone or the stimulatory effects of the kappa agonist butorphanol tartrate. It was hypothesized that the differential effects of ketocyclazocine in knife-cut and sham-operated rats are a consequence of the sedative effects of the drug combined with the elevated baseline of the knife-cut subjects. This hypothesis was supported by a third experiment, in which ketocyclazocine also reduced nocturnal intake in unoperated rats and butorphanol increased intake. That feeding responses to naloxone and butorphanol were essentially unchanged by hypothalamic knife cuts suggests that the opioid feeding system is independent of the longitudinal feeding inhibitory pathway believed to be involved in knife-cut-induced hyperphagia.


The Helicobacter pylori gene encoding phosphatidylserine synthase: sequence, expression, and insertional mutagenesis.
The Helicobacter pylori pss gene, coding for phosphatidylserine synthase (PSS), was cloned and sequenced in this study. A polypeptide of 237 amino acids was deduced from the PSS sequence. H. pylori PSS exhibits significant amino acid sequence identity with the PSS proteins found in the archaebacterium Methanococcus jannaschii, the gram-positive bacterium Bacillus subtilis, and the yeast Saccharomyces cerevisiae but none with its Escherichia coli counterpart. Expression of the putative pss gene in maxicells gave rise to a product of approximately 26 kDa, which is in agreement with the predicted molecular mass of 26,617 Da. A manganese-dependent PSS activity was found in the membrane fractions of the E. coli cells overexpressing the H. pylori pss gene product. This result indicates that this enzyme is a membrane-bound protein, a conclusion which is supported by the fact that the PSS protein contains several local hydrophobic segments which could form transmembrane helices. The pss gene was inactivated with a chloramphenicol acetyltransferase cassette on the plasmid. However, an isogenic pss gene-disrupted mutant of H. pylori UA802 could not be obtained, suggesting that this enzyme plays an essential role in the growth of this organism.


Effectivity of long antigen exposition dendritic cell therapy (LANEXDC®) in the palliative treatment of pancreatic cancer.
In pancreatic cancer median survival times range around 6, 6 to 6,9 months. Here we retrospectively analyzed the outcome of immunotherapy in the additional palliative treatment of pancreatic cancer with long antigen exposition dendritic cell therapy (LANEX-DC(®)) in 138 patients who were treated at our institution. Data were available of 134 patients (97.1%). The median interval between first diagnosis and start of treatment was 1.4 months. Therapy was well tolerated and no serious side effects were observed. The survival rate after 6 months was 72.2 % and afters 9 month 50.4%. The median survival time according to Kaplan- Meier regression analysis was 8.9 months. Median survival was significantly higher in the group of patients who started immunotherapy within 2 months following diagnosis (p=0.029) or repeated immunotherapy (p=0.027). Interestingly, younger patients <= 60 years of age lived significantly longer as patients > 60 years of age (p = 0.022). We were able to demonstrate in a large retrospective analysis that additional treatment with dendritic cells (LANEX-DC(®)) is highly effective and extends the median survival times up to 8.9 months. Furthermore we were able to demonstrate that median survival can be increased by early beginning and repetition of LANEX-DC(®) treatment.


Triamcinolone-induced systemic infections in female mice.
Six-week-old female mice were injected for prolonged periods of time with a synthetic glucocorticoid (triamcinolone diacetate). With the progression of the experiment the mortality rate increased significantly. Tissue samples were obtained from the surviving animals after 10, 22, 35 and 45 consecutive daily injections. Cultures were obtained from kidney and liver homogenates as well as from blood samples. Quantitative measurements indicated severe renal and hepatic infections which were followed by bactoeremia. The prevalent organisms in all cultures were enterobacteriae such as E. coli, Klebsiella, Pseudomonas and Streptococcus fecalis. Placebo-control animals which received daily injections of the vehicle revealed a moderate, transient infection.


The Management of Data for the Banking, Qualification, and Distribution of Induced Pluripotent Stem Cells: Lessons Learned from the European Bank for Induced Pluripotent Stem Cells.
The European Bank for induced pluripotent Stem Cells (EBiSC) was established in 2014 as a non-profit project for the banking, quality control, and distribution of human iPSC lines for research around the world. EBiSC iPSCs are deposited from diverse laboratories internationally and, hence, a key activity for EBiSC is standardising not only the iPSC lines themselves but also the data associated with them. This includes enabling unique nomenclature for the cells, as well as applying uniformity to the data provided by the cell line generator versus quality control data generated by EBiSC, and providing mechanisms to share personal data in a secure and GDPR-compliant manner. A joint approach implemented by EBiSC and the human pluripotent stem cell registry (hPSCreg®) has provided a solution that enabled hPSCreg® to improve its registration platform for iPSCs and EBiSC to have a pipeline for the import, standardisation, storage, and management of data associated with EBiSC iPSCs. In this work, we describe the experience of cell line data management for iPSC banking throughout the course of EBiSC's development as a central European banking infrastructure and present a model for how this could be implemented by other iPSC repositories to increase the FAIRness of iPSC research globally.


Tetrahydrobiopterin loading test in hyperphenylalaninemia.
Some cases of primary hyperphenylalaninemia are not caused by the lack of phenylalanine hydroxylase, but by the lack of its cofactor tetrahydrobiopterin. These patients are not clinically responsive to a phenylalanine-restricted diet, but need specific substitution therapy. Thus, it became necessary to examine all newborns screened as positive with the Guthrie test for tetrahydrobiopterin deficiency. Methods based on urinary pterin or on specific enzyme activity measurements are limited in their availability, and the simplest method, based on the lowering of serum phenylalanine after loading with cofactor, was discouraged by the finding that some dihydropteridine reductase-deficient patients were unresponsive. The preliminary observation that this limitation could be overcome by increasing the dose of the administered cofactor prompted us to reevaluate the potential of the tetrahydrobiopterin loading test in hyperphenylalaninemia. Fifteen patients, eight with ultimate diagnosis of phenylketonuria, three with 6-pyruvoyl tetrahydropterin synthase-, and four with dihydropteridine reductase-deficiency, have been examined by administering synthetic tetrahydrobiopterin both orally, at doses of 7.5 and 20 mg/kg, and i.v., at a dose of 2 mg/kg. All the tetrahydrobiopterin-deficient patients, unlike those with phenylketonuria, responded to the oral dose of 20 mg/kg cofactor by lowering their serum phenylalanine concentration markedly below baseline to an extent easily detectable by Guthrie cards. This method allows for a simple screening method when enzyme or pterin studies are not available.


Reversal of methotrexate inhibition of colony growth of L1210 leukemia cells in semisolid medium.
L1210 mouse leukemia cells were grown in a methylcellulose-based medium, and the inhibitory effect of methotrexate (MTX) on colony formation and its reversal were examined. The effect on colony formation was studied in order to compare the results with those obtained with normal mouse bone marrow cells grown in a similar manner in previous studies and in additional experiments presented. Light microscopy could not be used for colony counting of L1210 cells because MTX did not inhibit colony formation and only affected further colony growth. Therefore, it was necessary to evaluate the toxic effect of MTX by analysis of colony size distribution using an electronic image analyzer. Results show that the reversal of MTX toxicity to L1210 cells with leucovorin is competitive and is similar to that with normal mouse bone marrow cells. Thymidine in combination with a purine prevents MTX toxicity as well. The optimal concentration of thymidine is 10(-5) M, whereas at least 10(-4) M purine is required. Reversal of MTX toxicity by thymidine and purines is independent of the MTX concentration and is possible at drug concentrations as high as 10(-4) M. Compared to mouse bone marrow cells, L1210 cells appear to require more purines to prevent the toxic effects of MTX. MTX toxicity towards bone marrow myeloid precursor cells can be reversed by 10(-4) M inosine alone. These cells are better protected against MTX toxicity when 10(-6) to 10(-5) M thymidine is added. The results suggest that the use of a high-purine-low-thymidine combination has advantages over the use of leucovorin in controlling toxicity over a wide range of MTX concentrations and in providing some degree of selective protection to normal proliferating cells.


[Diagnostics and correction of malnutrition in gastrointestinal cancer].
Adequate nutrition plays a major role, both in prevention and treatment of gastrointestinal cancer. There are such factors as the cellulose content in food ration, the consumption of fresh fruits and vegetables, vitamins, physical activity. Cancer cachexia are observed at 70% of oncology patients in the terminal stage of cancer. Purposes of nutritional support are improvement of the quality of life, in particular terminally ill, the reduction of the length of hospital stay and costs of treatment, the prevention infectious and postoperative complications. The preference must be given enteral versus parenteral nutrition. In this work was studied possibilities of nutritional support for correction of malnutrition in the patient with gastrointestinal cancer. It is shown that the enteral nutrition contributes to improvement of nutritional status, prevents development of cachexia, improves the quality of life.


The bioengineering theory of the key modes of action of a cyanoacrylate liquid skin protectant.
The objective of this article is to formulate a new bioengineering theoretical framework for modelling the biomechanical efficacy of cyanoacrylate skin protectants, with specific focus on the Marathon technology (Medline Industries, Inc., Northfield, Illinois) and its modes of action. This work details the bioengineering and mathematical formulations of the theory, which is based on the classic engineering theories of flexural stiffness of coated elements and deformation friction. Based on the relevant skin anatomy and physiology, this paper demonstrates: (a) the contribution of the polymerised cyanoacrylate coating to flexural skin stiffness, which facilitates protection from non-axial (eg, compressive) localised mechanical forces; and (b) the contribution of the aforementioned coating to reduction in frictional forces and surface shear stresses applied by contacting objects such as medical devices. The present theoretical framework establishes that application of the cyanoacrylate coating provides considerable biomechanical protection to skin and subdermally, by shielding skin from both compressive and frictional (shearing) forces. Moreover, these analyses indicate that the prophylactic effects of the studied cyanoacrylate coating become particularly strong where the skin is thin or fragile (typically less than ~0.7 mm thick), which is characteristic to old age, post-neural injuries, neuromuscular diseases, and in disuse-induced tissue atrophy conditions.


Ghrelin and leptin pathophysiology in chronic kidney disease.
Ghrelin is an orexigenic hormone with additional effects on the regulation of inflammation and the cardiovascular system. It may play an important role in the pathogenesis of cachexia/protein-energy wasting (PEW), inflammation and cardiovascular complications in chronic kidney disease (CKD). There are three circulating gene products of ghrelin, namely, acyl ghrelin, des-acyl ghrelin and obestatin, each with individual distinct functions. Perturbations of these circulating ghrelin proteins impact the overall milieu of CKD. Leptin is an anorexigenic hormone which is secreted from the adipocytes and interacts with ghrelin and other appetite-regulating hormones. Leptin also plays a role in regulating inflammation and the cardiovascular system. Indeed, ghrelin and leptin may play yin-and-yang roles in CKD pathophysiology. Clinical trials involving the use of the mimetics or antagonists of these hormones are limited to short-term phase I/II studies. Further understanding of their interactions in CKD pathophysiology is needed for potential large-scale clinical trials, which may impact the quality of life and survival of patients with CKD.


Long-term treatment with oral sildenafil in addition to continuous IV epoprostenol in patients with pulmonary arterial hypertension.
To evaluate the effect of long-term oral therapy with sildenafil in patients with pulmonary arterial hypertension receiving long-term IV epoprostenol. Open, uncontrolled trial. University hospital. Two patients with primary pulmonary hypertension and one patient with pulmonary arterial hypertension after surgical closure of an atrial septal defect. All patients were receiving continuous epoprostenol for 1.7 to 7.1 years; two patients also received inhaled iloprost for 1.8 years and 3.8 years, respectively. Addition of oral sildenafil, up to 200 mg/d, divided in four to six single doses, and hemodynamic measurements and the 6-min walking distance (6MWD) before and after 5 months of treatment with sildenafil. One patient was treated with sildenafil, 200 mg/d; two patients received 75 mg/d due to nausea and headache. Long-term treatment with sildenafil in the three patients reduced mean pulmonary artery pressure by 14%, 41%, and 22%, respectively; in two patients, pulmonary vascular resistance was decreased by 52% and 55%. The 6MWD increased by 34%, 6%, and 29%, respectively. No significant systemic hypotension or decrease of arterial oxygen saturation was seen. Sildenafil therapy may be of benefit in patients with pulmonary arterial hypertension receiving long-term infusion of epoprostenol.


Setting the Stage: A Comparative Analysis of an Onstage/Offstage and a Linear Clinic Modules.
The purpose of this study was to understand how two different ambulatory design modules-traditional and onstage/offstage-impact operational efficiency, patient throughput, staff collaboration, and patient privacy. Delivery of healthcare is greatly shifting to ambulatory settings because of rapid advancement of medicine and technology, resulting in more day procedures and follow-up care occurring outside of hospitals. It is anticipated that outpatient services will grow roughly 15-23% within the next 10 years (Sg2, 2014). Nonetheless, there is limited research that evaluates how the built environment impacts care delivery and patient outcomes. This is a cross-sectional, comparative study consisted of a mixed-method approach that included shadowing clinic staff and observing and surveying patients. The linear module had shared corridors and publicly exposed workstations, whereas the onstage/offstage module separates patient/visitors from staff with dedicated patient corridors leading to exam rooms (onstage) and enclosed staff work cores (offstage). Roughly 35 hr of clinic staff shadowing and 55 hr of patient observations occurred. A total of 269 questionnaires were completed by patients/visitors. The results demonstrate that the onstage/offstage module significantly improved staff workflow, reduced travel distances, increased communication in private areas, and significantly reduced patient throughput and wait times. However, patients' perception of privacy did not change among the two modules. Compared to the linear module, this study provides evidence that the onstage/offstage module could have helped to optimize operational efficiencies, staff workflow, and patient throughput.


Diagnosis and Management of Anomalous Coronary Arteries with a Malignant Course.
Although the prevalence of anomalous coronary artery from the opposite sinus (ACAOS) in the general population is low, more frequent use of invasive and non-invasive imaging to rule out coronary artery disease has seen an increase in absolute numbers of ACAOS. ACAOS are traditionally classified as malignant (with an interarterial course) and benign variants. Malignant variants have been recognised in autopsy studies to be an underlying cause of sudden cardiac death in young athletes. Conversely, it seems that older people with ACAOS are less predisposed to adverse cardiac events. Non-invasive anatomic imaging is complementary to invasive imaging and helps to further identify high-risk anatomic features. Using functional non-invasive perfusion imaging can assess potential ischaemia induced by dynamic compression of malignant ACAOS. Information gained from clinical imaging guides the management of these patients.


Ten Important Considerations for Ovarian Cancer Screening.
The unique intricacies of ovarian cancer screening and perspectives of different screening methods are presented as ten considerations that are examined. Included in these considerations are: (1) Deciding on the number of individuals to be screened; (2) Anticipating screening group reductions due to death; (3) Deciding on the duration and frequency of screening; (4) Deciding on an appropriate follow-up period after screening; (5) Deciding on time to surgery when malignancy is suspected; (6) Deciding on how screen-detected ovarian cancers are treated and by whom; (7) Deciding on how to treat the data of enrolled participants; (8) Deciding on the most appropriate way to assign disease-specific death; (9) Deciding how to avoid biases caused by enrollments that attract participants with late-stage disease who are either symptomatic or disposed by factors that are genetic, environmental or social; and (10) Deciding whether the screening tool or a screening process is being tested. These considerations are presented in depth along with illustrations of how they impact the outcomes of ovarian cancer screening. The considerations presented provide alternative explanations of effects that have an important bearing on interpreting ovarian screening outcomes.


Cancer epidemiology and control in peninsular and island South-East Asia - past, present and future.
Malaysia, Brunei, Singapore, Indonesia, East Timor and the Philippines constitute peninsular and island South-East Asia. For reasons of largely shared ethnicity, with Chinese elements added to the basic Austromalaysian populations, as well as geographical contiguity, they can be usefully grouped together for studies of chronic disease prevalence and underlying risk factors. The fact of problems are shared in common, particularly regarding increasing cancer rates, underlines the necessity for a coordinated approach to research and development of control measures. To provide a knowledge base, the present review of available data for cancer registration, epidemiology and control was conducted. The most prevalent cancer site in males is the lung, followed by the liver, colon or the prostate in the majority of cases, while breast and cervical cancers predominate in most female populations. However, there are interesting differences among the racial groups, particularly regarding the stomach. General tendencies for increase in adenocarcinomas but decrease in squamous cell carcinomas and gastric cancer, point to change in environmental influence over time. Variation in risk factors depends to some extent on the level of economic development but overall the countries of the region face similar challenges in achieving effective cancer control. A major task is persuading the general populace of the efficacy of early detection and clinical treatment.


Copepod parasites of the fatheads (pisces, psychrolutidae) and their implication on the phylogenetic relationships of psychrolutid genera.
Nine species of psychrolutids kept in the Marine Zoology of Hokkaido University were found to carry eight species of copepod parasites. The parasites and their hosts are: Bobkabata kabatabobbus Hogans and Benz, 1990 on Malacocottus zonurus Bean; Chondracanthus parvus n. sp. on Eurymen gyrinus Gilbert and Burke; Chondracanthus yabei n. sp. on Dasycottus setiger Bean and M. zonurus; Ch. yanezi Atria, 1980 on Psychrolutes phrictus Stein and Bond; Caligus similis n. sp. on Neophrynichthys latus (Hutton); Clavella adunca (Strøm, 1762) on M. zonurus; Neobrachiella amphipacifica Ho, 1982 on Ambophthalmos angustus (Nelson), Cottunculus sp., D. setiger, Ebinania brephocephala (Jordan and Starks), E. vermiculata Sakamoto, and P. phrictus; and Naobranchia occidentalis Wilson, 1915 on D. setiger and M. zonurus. Chondracanthus parvus is closest to Ch. deltoideus Fraser, 1920, but differs from it in having only one pair of small knobs on the head and carrying a pair of lateral processes on the second pediger. Chondracanthus yabei resembles Ch. yanezi Atria, 1980, but can be distinguished from it by the presence of three, low protrusions on the mid-dorsal surface of the trunk; besides, maxillule also shows difference. As the name indicates, C. similis resembles several species of Caligus that bear a short abdomen and a formula of I; IV on the exopod of leg 4. However, it can be separated from them by the combination of the following characters: a genital complex distinctly smaller than the cephalothoracic shield, a pair of relatively short caudal rami, a smooth dentiform process on maxillule, and a pair of truncate tines on sternal furca. Analysis of the occurrence of Chondracanthus on the psychrolutid fishes shows that the phylogeny of Chondracanthus is in congruence with that of the Psychrolutidae. It implies that parasitism of Chondracanthus occurred after the Dasycottus clade diverged off the main stock of the Psychrolutidae and the occurrence of Ch. yabei on D. setiger is resulted from a later colonization.


Ferulic acid enhances the radiation sensitivity of lung and liver carcinoma cells by collapsing redox homeostasis: mechanistic involvement of Akt/p38 MAPK signalling pathway.
The major drawback of anticancer therapy is the development of resistance against drugs and radiation at the later phase of treatment which may lead to recurrences of the disease. Therefore, strategy was taken to enhance radiation sensitivity of lung (A549) and liver (HepG2) carcinoma cells by treatment with ferulic acid (FA) prior to irradiation. FA pre-treatment initially decreased reactive oxygen species (ROS) level in carcinoma cells which induced reductive stress and cytostasis. To overcome this stress, cellular mechanism increased the Keap1 level to down-regulate nuclear localisation of Nrf2 and its dependent antioxidant system. The antioxidant system reached the lowest level after 3 and 6 h of FA treatment in A549 and HepG2 cells respectively. As endogenous ROS were still being generated at same rate, ROS level was clearly higher than control which changed the reductive stress to oxidative stress. Exposure to γ-radiation in this condition further increased ROS level and caused radio-sensitisation of carcinoma cells. Combination of irradiation (IR) and FA activated mitochondrial apoptotic pathway and concomitantly inhibited the cell cycle progression and survival pathway over the IR group. Moreover, the combination treatment showed significant tumour regression, caspase 3 activation and nuclear fragmentation in tumour tissue compared to radiation alone. In contrast, FA pre-treatment protected peripheral blood mononuclear cells (PBMC) and normal lung fibroblast WI38 cells from radiation damage. Together, combination treatment offers effective strategy of killing cancer cells and demonstrates its potential for increasing the efficacy of radio-therapy.


Liquid liothyronine to obtain target TSH in differentiated thyroid cancer patients.
In the last ten years a liquid formulation of liothyronine (L-T3) became available. To date, no studies on its systematic use have been reported. This study is aimed at assessing the reliability of liquid L-T3 in achieving target TSH in patients with differentiated thyroid cancers (DTC). Twenty-one high risk DTC patients in whom levothyroxine treatment up to 2.0 μg/kg/day did not suppress TSH levels (i.e. >0.1 mIU/L) were selected. Maintaining the same L-T4 dose, they started to assume liquid L-T3 at an initial fixed dose of 3.55 μg (5 drops). Further adjustments of L-T3 dose were tailored according to individual assessment. Initial serum TSH ranged from 0.8 to 12.0 mIU/L, when patients assumed high dose of L-T4 alone. Following the addition of a daily single dose of 3.55 μg L-T3, the target TSH was attained in five patients (23.8%). After increasing L-T3 dose up to a mean of 7.3±3.4 μg/day all patients reached target serum TSH (<0.1 mIU/L). The mean individual L-T3 dose was significantly correlated with the body weight and was 0.11±0.04 μg/kg/day (p=0.013). Mean L-T4:L-T3 ratio was 21:1. No patients showed skewed free-T3 or free-T4 values, neither experienced discomfort nor reported adverse events. Liquid L-T3 can be useful to achieve optimal TSH suppression in high risk DTC with not suppressed TSH on L-T4 alone. This formulation allows an individual tailoring of L-T3, minimizing risks of side effects as well as of overtreatment in these clinical conditions.


Transient adhesion refractoriness of circulating platelets under shear stress: the role of partial activation and microaggregate formation by suboptimal ADP concentration.
Exposure of whole blood (WB) to subendothelial extracellular matrix (ECM) under shear stress in the cone and plate(let) analyser (CPA) results in platelet adhesion, followed by release reaction and aggregation of circulating platelets on the adherent platelets. The properties of circulating non-adhered platelets in the CPA was studied by exposure of WB to ECM at a high shear rate (1300/s) for 2 min (1st run), followed by transfer of the suspension to a new ECM-coated well for a second run (2nd run) under similar conditions. The results of the 2nd run demonstrated transient adhesion refractoriness associated with platelet microaggregate formation in the suspension. The adhesion refractoriness was dependent on platelet activation during the 1st run and was prevented by addition of apyrase (ADP scavenger) or ADP receptor inhibitor, suggesting a role for ADP in mediating this response. Furthermore, exposure of WB samples to suboptimal concentrations of ADP (0.4-1 micromol/l) or a thrombin receptor activating peptide (TRAP) (5 micromol/l) for 2 min resulted in a similar transient platelet adhesion refractoriness to ECM under flow conditions. The transient platelet refractoriness and microaggregate formation induced by ADP was associated with a transient reduction in glycoprotein (GP)Ib, increased P-selectin expression and increased fibrinogen binding by circulating platelets. These data suggest a role for platelet agonists at suboptimal concentrations in modulating platelet function and limiting the expansion of the thrombus.


Is Single-incision Laparoscopic Cholecystectomy Feasible for Acute Cholecystitis? A Consecutive Study of 60 Cases.
The feasibility of single-incision laparoscopic cholecystectomy (SIL-C) for patients with acute cholecystitis were evaluated based on the timing of operation after onset of symptoms. Sixty patients with acute cholecystitis who underwent SIL-C were divided into 2 groups according to the timing of operation: group E included 23 patients who underwent SIL-C within 72 hours, and group O included 37 patients who underwent SIL-C later. There were no statistical differences between group E and group O in clinicopathologic characteristics. Group E demonstrated significantly shorter operating time and less blood loss than group O. Although the incidences of additional port(s) requirements were not significantly different, 8 patients in group O required open conversion, indicating significantly higher rate. As group E demonstrated favorable surgical outcomes compared with group O, SIL-C for acute cholecystitis seems to be a feasible therapeutic procedure when performed within 72 hours as updated Tokyo Guidelines recommended.


Dietary intake of one-carbon metabolism nutrients and DNA methylation in peripheral blood.
Folate and other one-carbon metabolism nutrients are essential to enable DNA methylation to occur, but the extent to which their dietary intake influences methylation in adulthood is unclear. We assessed associations between dietary intake of these nutrients and DNA methylation in peripheral blood, overall and at specific genomic locations. We conducted a cross-sectional study using baseline data and samples from 5186 adult participants in the Melbourne Collaborative Cohort Study (MCCS). Nutrient intake was estimated from a food-frequency questionnaire. DNA methylation was measured by using the Illumina Infinium HumanMethylation450 BeadChip array (HM450K). We assessed associations of intakes of folate, riboflavin, vitamins B-6 and B-12, methionine, choline, and betaine with methylation at individual cytosine-guanine dinucleotides (CpGs), and with median (genome-wide) methylation across all CpGs, CpGs in gene bodies, and CpGs in gene promoters. We also assessed associations with methylation at long interspersed nuclear element 1 (LINE-1), satellite 2 (Sat2), and Arthrobacter luteus restriction endonuclease (Alu) repetitive elements for a subset of participants. We used linear mixed regression, adjusting for age, sex, country of birth, smoking, energy intake from food, alcohol intake, Mediterranean diet score, and batch effects to assess log-linear associations with dietary intake of each nutrient. In secondary analyses, we assessed associations with low or high intakes defined by extreme quintiles. No evidence of log-linear association was observed at P < 10-7 between the intake of one-carbon metabolism nutrients and methylation at individual CpGs. Low intake of riboflavin was associated with higher methylation at CpG cg21230392 in the first exon of PROM1 (P = 5.0 × 10-8). No consistent evidence of association was observed with genome-wide or repetitive element measures of methylation. Our findings suggest that dietary intake of one-carbon metabolism nutrients in adulthood, as measured by a food-frequency questionnaire, has little association with blood DNA methylation. An association with low intake of riboflavin requires replication in independent cohorts. This study was registered at http://www.clinicaltrials.gov as NCT03227003.