Correlation of cell migration, cell invasion, receptor number, proteinase production, and basic fibroblast growth factor levels in endothelial cells.
The levels of endogenous basic fibroblast growth factor (bFGF) in seven clones of cultured bovine capillary endothelial (BCE) cells were assayed, and their relation to cell morphology, bFGF receptor number, cell migration, amniotic membrane invasivity, and proteinase levels were studied. Immunoblotting experiments with anti-bFGF IgG demonstrated that cells from these clones contained different amounts of bFGF. The cells containing high levels of bFGF had a spindle or elongated appearance at confluence and a low number of high affinity receptors for bFGF. The cells containing low levels of bFGF had a cobblestone-like appearance and a higher number of high affinity receptors. When exposed to 10 ng/ml bFGF, cells containing a low level of bFGF took on an elongated appearance with a crisscross pattern similar to that seen with the high producer bFGF cells. The endogenous bFGF levels of the BCE cell clones correlated with the extent of cell migration after wounding of a monolayer and the degree of invasion of the human amniotic membrane. Cells from the clone with the highest endogenous bFGF level migrated well, invaded the amnion membrane without the addition of exogenous bFGF, and were relatively unaffected by the addition of bFGF. Cells from the clone containing the lowest level of bFGF did not migrate or invade under normal conditions. However, the addition of bFGF to the culture medium strongly enhanced both of these processes. The inclusion of anti-bFGF IgG in the media suppressed cell migration and invasion. The plasminogen activator (PA) activities of cell lysates of the clones, assayed by the 125I-fibrin plate technique, indicated that the PA levels did not correlate with the bFGF levels. Metalloproteinase activities in the conditioned medium, assayed by gelatin zymography, correlated with the endogenous bFGF levels, suggesting that the degree of expression of metalloproteinases might be critical for cell migration and invasion. These data suggest that endogenous bFGF may have an important role for migration and invasion of BCE cells during neovascularization via the induction and/or activation of specific metalloproteinases.


Reflections on discrimination and health in India.
This is a speculative paper on the structure of caste-based discrimination in India. It sketches the field by a) proposing four empirical and historical examples of discrimination in different medical situations; b) suggesting an analytical framework composed of domain, register, temporality and intensity of discrimination; c) proposing that in the Indian historical context, discrimination masks itself, hiding its character behind the veneer of secular ideas; d) arguing that discrimination is not some unfortunate residue of backwardness in modern society that will go away, but is the force of social hierarchy transforming itself into a fully modern capitalist culture. The paper then arrives at the understanding that discrimination is pandemic across India. The conclusion suggests that in India today, we need proposals, hypotheses and arguments that help us establish the ethical framework for meaningful empirical research that sociological studies of medical ethics and the epidemiology of discrimination can pursue. Its method is that of logical and speculative argument based on experience, with examples of different forms of discrimination to clarify the point being made. No specific research was undertaken for this purpose since the paper is not empirically based.


Monopodal Postural Stability Assessment by Wireless Inertial Measurement Units Through the Fast Fourier Transform.
(1) To describe the fast Fourier transform (FFT) multijoint as monopodal postural stability measurement in well-trained athletes, (2) to compare the within-subject FFT between laterality, joints, and body segments, and (3) to establish the within- and between-subject relationship between joints. Twelve national-level basketball players participated voluntarily in this investigation. The participants performed two 60-second repetitions of a monopodal stability test (1 repetition with each lower limb), separated by 3 minutes of active recovery. All tests were recorded by 4 WIMU PRO™ inertial devices located on the ankle, knee, lumbar spine, and thoracic spine. The main variable was total acceleration, where the FFT was applied. The higher instability results were found in the ankle and in the nondominant lower limb (dominant = 1.131 [0.122] a.u. (arbitrary units); nondominant = 1.141 [0.172] a.u). In the body segment analysis, the greater percentage of differences (%diff) were shown between lumbar spine and knee in the dominant (%diff = -2.989%; d = 0.87) and nondominant (%diff = -3.243%; d = 0.90) lower limb. Finally, very large between-subjects variability was found in all joints and body segments. The described protocol is proposed for monopodal postural stability assessment, being useful to provide information about the stability of joints and the body segment between joints. Besides, a within-subject analysis is recommended, and the FFT calculation will enable a linear analysis of each test.


Bioactive pyranoxanthones from the roots of Calophyllum blancoi.
Phytochemical investigation of the roots of Calophyllum blancoi growing in Taiwan resulted in the isolation of three new pyranoxanthones, blancoxanthone (1), acetyl blancoxanthone (2) and 3-hydroxyblancoxanthone (3), in addition to two known pyranoxanthones, pyranojacaeubin (4) and caloxanthone (5). Structural characterization of the isolated compounds was determined by spectral analyses especially 2-D NMR. Biological study of the isolated xanthones revealed that blancoxanthone (1) exhibited significant anti-coronavirus activity.


Non-AIDS-defining deaths and immunodeficiency in the era of combination antiretroviral therapy.
To assess whether immunodeficiency is associated with the most frequent non-AIDS-defining causes of death in the era of combination antiretroviral therapy (cART). Observational multicentre cohorts. Twenty-three cohorts of adults with estimated dates of human immunodeficiency virus (HIV) seroconversion were considered. Patients were seroconverters followed within the cART era. Measurements were latest CD4, nadir CD4 and time spent with CD4 cell count less than 350 cells/microl. Outcomes were specific causes of death using a standardized classification. Among 9858 patients (71 230 person-years follow-up), 597 died, 333 (55.7%) from non-AIDS-defining causes. Non-AIDS-defining infection, liver disease, non-AIDS-defining malignancy and cardiovascular disease accounted for 53% of non-AIDS deaths. For each 100 cells/microl increment in the latest CD4 cell count, we found a 64% (95% confidence interval 58-69%) reduction in risk of death from AIDS-defining causes and significant reductions in death from non-AIDS infections (32, 18-44%), end-stage liver disease (33, 18-46%) and non-AIDS malignancies (34, 21-45%). Non-AIDS-defining causes of death were also associated with nadir CD4 while being cART-naive or duration of exposure to immunosuppression. No relationship between risk of death from cardiovascular disease and CD4 cell count was found though there was a raised risk associated with elevated HIV RNA. In the cART era, the most frequent non-AIDS-defining causes of death are associated with immunodeficiency, only cardiovascular disease was associated with high viral replication. Avoiding profound and mild immunodeficiency, through earlier initiation of cART, may impact on morbidity and mortality of HIV-infected patients.


Placental syncytiotrophoblast constitutes a major barrier to vertical transmission of Listeria monocytogenes.
Listeria monocytogenes is an important cause of maternal-fetal infections and serves as a model organism to study these important but poorly understood events. L. monocytogenes can infect non-phagocytic cells by two means: direct invasion and cell-to-cell spread. The relative contribution of each method to placental infection is controversial, as is the anatomical site of invasion. Here, we report for the first time the use of first trimester placental organ cultures to quantitatively analyze L. monocytogenes infection of the human placenta. Contrary to previous reports, we found that the syncytiotrophoblast, which constitutes most of the placental surface and is bathed in maternal blood, was highly resistant to L. monocytogenes infection by either internalin-mediated invasion or cell-to-cell spread. Instead, extravillous cytotrophoblasts-which anchor the placenta in the decidua (uterine lining) and abundantly express E-cadherin-served as the primary portal of entry for L. monocytogenes from both extracellular and intracellular compartments. Subsequent bacterial dissemination to the villous stroma, where fetal capillaries are found, was hampered by further cellular and histological barriers. Our study suggests the placenta has evolved multiple mechanisms to resist pathogen infection, especially from maternal blood. These findings provide a novel explanation why almost all placental pathogens have intracellular life cycles: they may need maternal cells to reach the decidua and infect the placenta.


Knee arthrodesis for failed total knee arthroplasty.
The use of arthrodesis to salvage failed total knee arthroplasty is reviewed in nine patients. Prosthetic failure was due to infection in six cases, aseptic loosening in two cases, and instability in one case. The techniques of arthrodesis included six Hoffman external fixators, two fluted intramedullary rods (3M-Orthopedic Products Division, St Paul), and one pulsing electromagnetic field stimulator. Arthrodesis was successful in eight of nine patients, averaging 5.4 months to union. All eight patients resumed a painfree functional level of activity. Meticulous surgical technique and appropriate method of arthrodesis are emphasized.


Substrate profile in rat soleus muscle fibers after hindlimb unloading and fatigue.
Limb muscles from rats flown in space and after hindlimb unloading (HU) show an increased fatigability, and spaceflight has been shown to result in a reduced ability to oxidize fatty acids. The purpose of this investigation was to determine the effects of HU on the substrate content in fast- and slow-twitch fibers and to assess the substrate utilization patterns in single slow type I fibers isolated from control and HU animals. A second objective was to assess whether HU altered the ability of the heart or limb muscle to oxidize pyruvate or palmitate. After 2 wk of HU, single fibers were isolated from the freeze-dried soleus and gastrocnemius muscles. HU increased the glycogen content in all fiber types, and it increased lactate, ATP, and phosphocreatine in the slow type I fiber. After HU, the type I fiber substrate profile was shifted toward that observed in fast fibers. For example, fiber glycogen increased from 179 +/- 16 to 285 +/- 25 mmol/kg dry wt, which approached the 308 +/- 23 mmol/kg dry wt content observed in the post-HU type IIa fiber. With contractile activity, the type I fiber from the HU animal showed a greater utilization of glycogen and accumulation of lactate compared with the control type I fiber. HU had no effect on the ability of crude homogenate or mitochondria fractions from the soleus or gastrocnemius to oxidize pyruvate or palmitate. The increased fatigability after HU may have resulted from an elevated glycolysis producing an increased cell lactate and a decreased pH.


Association of Higher Consumption of Foods Derived From Subsidized Commodities With Adverse Cardiometabolic Risk Among US Adults.
Food subsidies are designed to enhance food availability, but whether they promote cardiometabolic health is unclear. To investigate whether higher consumption of foods derived from subsidized food commodities is associated with adverse cardiometabolic risk among US adults. Cross-sectional analysis of the National Health and Nutrition Examination Survey data from 2001 to 2006. Our final analysis was performed in January 2016. Participants were 10 308 nonpregnant adults 18 to 64 years old in the general community. From a single day of 24-hour dietary recall in the National Health and Nutrition Examination Survey, we calculated an individual-level subsidy score that estimated an individual's consumption of subsidized food commodities as a percentage of total caloric intake. The main outcomes were body mass index (calculated as weight in kilograms divided by height in meters squared), abdominal adiposity, C-reactive protein level, blood pressure, non-high-density lipoprotein cholesterol level, and glycemia. Among 10 308 participants, the mean (SD) age was 40.2 (0.3) years, and a mean (SD) of 50.5% (0.5%) were male. Overall, 56.2% of calories consumed were from the major subsidized food commodities. United States adults in the highest quartile of the subsidy score (compared with the lowest) had increased probabilities of having a body mass index of at least 30 (prevalence ratio, 1.37; 95% CI, 1.23-1.52), a ratio of waist circumference to height of at least 0.60 (prevalence ratio, 1.41; 95% CI, 1.25-1.59), a C-reactive protein level of at least 0.32 mg/dL (prevalence ratio, 1.34; 95% CI, 1.19-1.51), an elevated non-high-density lipoprotein cholesterol level (prevalence ratio, 1.14; 95% CI, 1.05-1.25), and dysglycemia (prevalence ratio, 1.21; 95% CI, 1.04-1.40). There was no statistically significant association between the subsidy score and blood pressure. Among US adults, higher consumption of calories from subsidized food commodities was associated with a greater probability of some cardiometabolic risks. Better alignment of agricultural and nutritional policies may potentially improve population health.


Ecological study of Streptococcus mutans, Streptococcus sobrinus and Lactobacillus spp. at sub-sites from approximal dental plaque from children.
Previous immunofluorescence (IF) studies have indicated that Streptococcus mutans may preferentially colonise specific sub-sites within approximal plaque. The present study aimed to extend these observations to other mutans streptococci and lactobacilli in such gingival margin plaque. Two hundred and seventy approximal plaque samples were taken from 90 teeth (3 from each tooth) in 64 children; three gingival margin sub-sites in relation to the contact area: away from (A), to the side of(S) and below (B) the contact area. Samples were processed by indirect IF using high-titred polyclonal anti-S. mutans 'c', anti-S. sobrinus 'd', anti-L. casei and anti-L. acidophilus antisera. An overall positive association was found between S. mutans 'c' and S. sobrinus 'd' (p < 0.001). Significant differences (p < 0.1) were found between the proportional counts at each sub-site for S. mutans 'c': A = 39%, S = 51% and B = 70%, and for S. sobrinus 'd' 21, 33 and 49%. Mutans streptococci (MS) appeared to preferentially colonise the sub-site below the contact area (B = 81%), compared with sub-sites A and S (48 and 62%, respectively). S. mutans 'c' and S. sobrinus 'd' were detected together at subsites A = 12%, S = 22%, and B = 38%, with proportional counts at B sites being higher than those at A (B > A, p < 0.01, and B > S, p < 0.05). Lactobacillus spp. were isolated rarely, and were usually found together with MS. There was a positive relationship between the presence of lactobacilli or MS and caries (white spot lesions only), although these species could frequently be isolated from noncarious sites. The presence of both S. mutans 'c' and S. sobrinus 'd' were strongly correlated with early caries lesions. In addition, this study confirmed the variation in the microflora at different sub-sites within approximal dental plaque.


Structural and functional insights into the roles of the Mms21 subunit of the Smc5/6 complex.
The Smc5/6 complex is an evolutionarily conserved chromosomal ATPase required for cell growth and DNA repair. Its Mms21 subunit supports both functions by docking to the arm region of Smc5 and providing SUMO ligase activity. Here, we report the crystal structure of Mms21 in complex with the Smc5 arm. Our structure revealed two distinct structural and functional domains of the Smc5-bound Mms21: its N-terminal half is dedicated to Smc5 binding by forming a helix bundle with a coiled-coil structure of Smc5; its C-terminal half includes the SUMO ligase domain, which adopts a new type of RING E3 structure. Mutagenesis and structural analyses showed that the Mms21-Smc5 interface is required for cell growth and resistance to DNA damage, while the unique Mms21 RING domain confers specificity to the SUMO E2-E3 interaction. Through structure-based dissection of Mms21 functions, our studies establish a framework for understanding its roles in the Smc5/6 complex.


The moderating role of centrality on associations between ethnic identity affirmation and ethnic minority college students' mental health.
Prior literature has shown that ethnic affirmation, one aspect of ethnic identity, is positively associated with mental health. However, the associations between ethnic affirmation and mental health may vary depending how much importance individuals place on their ethnic group membership (ie, centrality). Using path analysis, the current study examined the relations between ethnic affirmation and indices of mental health problems (ie, anxiety and depressive symptoms), and tested whether the process was moderated by ethnic centrality among 3,659 college students representing 3 ethnic groups (41% Latino/a, 35% Asian American, and 24% African American) who participated in a large, multisite university study. Results suggested that the associations between ethnic affirmation and mental health were stronger for Latino/a and Asian American students who reported higher levels of ethnic centrality. For African Americans, higher levels of ethnic affirmation predicted better mental health, but this association did not vary as a function of ethnic centrality.


SV40 activates transcription from the transferrin receptor promoter by inducing a factor which binds to the CRE/AP-1 recognition sequence.
During the course of lytic infection by simian virus 40 (SV40), expression of both the viral late genes and certain host cellular genes is induced. The promoter of the cellular transferrin receptor (TR) gene contains a DNA sequence which is similar to the AP-1- and AP-4-binding region in SV40 which has been implicated in the control of the viral late promoter. Expression of TR is needed for cells to enter S-phase and is therefore expected to be important for the SV40 lytic cycle. Here we show that the level of TR mRNA in vivo was increased by SV40 infection. A factor which activates transcription from the TR promoter in vitro was specifically induced in SV40-infected cells. Gel mobility shift assays with an oligonucleotide comprising this part of the TR promoter showed three nucleoprotein complexes to be formed with proteins from CV-1 cells. Following SV40 infection, one of the complexes was increased ten-fold. Formation of this complex was specifically reduced by competition with the phorbol ester-responsive element of the collagenase gene, implying that the factor is a member of the AP-1/Jun/Fos family. Cross-linking of the complex by ultraviolet light showed major DNA-binding components to be proteins of about 55 kD and 47 kD. Removal of this factor by adding the oligonucleotide to in vitro transcription reactions with the TR promoter, abolished the activation of TR transcription. The factor which binds to the TR promoter co-sedimented with SV40 chromosomes extracted late in infection. This suggests that similar transcriptional regulatory proteins are involved in controlling transcription from both the SV40 and the TR promoters, and that the virus can use a common mechanism to induce viral and host cellular transcription.


Identification of ten serum microRNAs from a genome-wide serum microRNA expression profile as novel noninvasive biomarkers for nonsmall cell lung cancer diagnosis.
The detection of nonsmall cell lung cancer (NSCLC) at an early stage presents a daunting challenge due to the lack of a specific noninvasive marker. The discovery of microRNAs (miRNAs), particularly those found in serum, has opened a new avenue for tumor diagnosis. To determine whether the expression profile of serum miRNAs can serve as a NSCLC fingerprint, we performed Taqman probe-based quantitative RT-PCR assay to selected differentially expressed serum miRNAs from a sample set including 400 NSCLC cases and 220 controls, and risk score analysis to evaluate the diagnostic value of the serum miRNA profiling system. After a two-phase selection and validation process, 10 miRNAs were found to have significantly different expression levels in NSCLC serum samples compared with the control serum samples. Risk score analysis showed that this panel of miRNAs was able to distinguish NSCLC cases from controls with high sensitivity and specificity. Under ROC curves, the AUC for tumor identification in training set and validation set were 0.966 and 0.972, respectively. Furthermore, the expression profile of the 10-serum miRNAs was correlated with the stage of NSCLC patients, especially in younger patients and patients with current smoking habits. More importantly, the serum miRNA-based biomarker for early NSCLC detection was supported by a retrospective analysis in which the 10-serum miRNA profile could accurately classify serum samples collected up to 33 months ahead of the clinical NSCLC diagnosis. Taken together, we demonstrate that the profiling of 10-serum miRNAs provides a novel noninvasive biomarker for NSCLC diagnosis.


Influence of load cycling on marginal microleakage with two self-etching and two one-bottle dentin adhesive systems in dentin.
To evaluate the influence of occlusal load cycling on cervical microleakage of proximal slot restorations located in dentin, using two self-etching and two one-bottle dentin adhesive systems. 240 proximal slot cavities were prepared in 120 bovine teeth and divided into two groups, one with load cycling and one without. The groups were then subdivided into four subgroups according to the adhesive system used (Experimental EXL 547 Self-etching 3M, Clearfil SE Bond, Single Bond, and Optibond Solo Plus) and restored following the manufacturers' instructions. The teeth were then submitted to mechanical load cycling with a force of 80 N and a frequency of 5 Hz, simultaneously over both restorations of each tooth, for a total of 50,000 cycles per specimen. All specimens were subsequently immersed in a 2% methylene blue solution (pH 7.0), and sectioned to examine the extent of dye penetration under a stereomicroscope (40X). There was no statistically significant difference (p = 0.00002) between the loaded and unloaded teeth. However, a statistically significant difference was observed between the adhesive systems used. The experimental self-etching EXL 547 presented the lowest mean microleakage, but was only statistically significantly different from the Single Bond loaded and unloaded groups and the Clearfil SE Bond unloaded group. The application of 50,000 loading cycles did not affect the microleakage of the two self-etching and the two one-bottle adhesive systems evaluated. In vitro mechanical load cycling is an important factor to consider when evaluating the performance of adhesive systems under simulated masticatory conditions.


New Seeding Approach Reduces Costs and Time to Outplant Sexually Propagated Corals for Reef Restoration.
The use of sexually propagated corals is gaining popularity as an approach for reef restoration. However, manually attaching substrates with recently settled corals to the reef using binding materials is both time-consuming and expensive, limiting the use of this technique to small spatial scales. We present a novel approach whereby young corals are 'seeded' on the reef without the need for manual attachment to the benthos. We tested two tetrapod-shaped concrete substrates (7.9 and 9.8 cm in diameter) on which coral larvae were settled. The tetrapods were efficiently deployed by wedging them in reef crevices, in 1.5 to 7% of the time required for traditional outplanting techniques. Seeding tetrapods was most effective in reefs with moderately to highly complex topographies, where they rapidly became lodged in crevices or cemented to the benthos by encrusting organisms. After one year, average recruit survival was 9.6% and 67% of tetrapods still harboured at least one coral colony, and overall, this approach resulted in a 5 to 18 fold reduction in outplanting costs compared to common outplanting methods. This seeding approach represents a substantial reduction in costs and time required to introduce sexually propagated corals to reefs, and could possibly enable larger scale reef restoration.


Effects of plant size and water relations on gas exchange and growth of the desert shrub Larrea tridentata.
Larrea tridentata is a xerophytic evergreen shrub, dominant in the arid regions of the southwestern United States. We examined relationships between gasexchange characteristics, plant and soil water relations, and growth responses of large versus small shrubs of L. tridentata over the course of a summer growing season in the Chihuahuan Desert of southern New Mexico, USA. The soil wetting front did not reach 0.6 m, and soils at depths of 0.6 and 0.9 m remained dry throughout the summer, suggesting that L. tridentata extracts water largely from soil near the surface. Surface soil layers (<0.3 m) were drier under large plants, but predawn xylem water potentials were similar for both plant sizes suggesting some access to deeper soil moisture reserves by large plants. Stem elongation rates were about 40% less in large, reproductively active shrubs than in small, reproductively inactive shrubs. Maximal net photosynthetic rates (Pmax) occurred in early summer (21.3 μ mol m-2 s-1), when pre-dawn xylem water potential (XWP) reached ca. -1 MPa. Although both shrub sizes exhibited similar responses to environmental factors, small shrubs recovered faster from short-term drought, when pre-dawn XWP reached about -4.5 MPa and Pmax decreased to only ca. 20% of unstressed levels. Gas exchange measurements yielded a strong relationship between stomatal conductance and photosynthesis, and the relationship between leaf-to-air vapor pressure deficit and stomatal conductance was found to be influenced by pre-dawn XWP. Our results indicate that stomatal responses to water stress and vapor pressure deficit are important in determining rates of carbon gain and water loss in L. tridentata.


Mechanical Properties, Quantum Mechanical Calculations, and Crystallographic/Spectroscopic Characterization of GaNbO4, Ga(Ta,Nb)O4, and GaTaO4.
Single crystals as well as polycrystalline samples of GaNbO4, Ga(Ta,Nb)O4, and GaTaO4 were grown from the melt and by solid-state reactions, respectively, at various temperatures between 1698 and 1983 K. The chemical composition of the crystals was confirmed by wavelength-dispersive electron microprobe analysis, and the crystal structures were determined by single-crystal X-ray diffraction. In addition, a high-P-T synthesis of GaNbO4 was performed at a pressure of 2 GPa and a temperature of 1273 K. Raman spectroscopy of all compounds as well as Rietveld refinement analysis of the powder X-ray diffraction pattern of GaNbO4 were carried out to complement the structural investigations. Density functional theory (DFT) calculations enabled the assignment of the Raman bands to specific vibrational modes within the structure of GaNbO4. To determine the hardness (H) and elastic moduli (E) of the compounds, nanoindentation experiments have been performed with a Berkovich diamond indenter tip. Analyses of the load-displacement curves resulted in a high hardness of H = 11.9 ± 0.6 GPa and a reduced elastic modulus of Er = 202 ± 9 GPa for GaTaO4. GaNbO4 showed a lower hardness of H = 9.6 ± 0.5 GPa and a reduced elastic modulus of Er = 168 ± 5 GPa. Spectroscopic ellipsometry of the polished GaTa0.5Nb0.5O4 ceramic sample was employed for the determination of the optical constants n and k. GaTa0.5Nb0.5O4 exhibits a high average refractive index of nD = 2.20, at λ = 589 nm. Furthermore, in situ high-temperature powder X-ray diffraction experiments enabled the study of the thermal expansion tensors of GaTaO4 and GaNbO4, as well as the ability to relate them with structural features.


Cumulative scales for the measurement of agoraphobia.
The questionnaire responses of 559 agoraphobic subjects were analysed to see whether there was a shared hierarchical pattern of fear and avoidance. The items were divided on the basis of a principal component analysis into 13 loading on an Agoraphobia factor and eight on a Claustrophobia factor. When analysed with Guttman Scaling Analysis valid cumulative scales were found for each factor. These scales were replicated in three independent cohorts of subjects and met the most stringent scaling requirements. The scales produced three valid scores of severity for each individual, the first describing exactly which Agoraphobic items the subject avoids, the second describing Agoraphobic items feared and the third Claustrophobic items avoided. Since it is possible to deduce the current pattern of fear and avoidance from the scores, they can be used to summarize the clinical state of an individual client or for making precise descriptive comparisons between agoraphobics . The scales also indicate that agoraphobics do not develop their own unique set of difficulties from the possible pool of problems, but acquire an ordered set of difficulties, the order being shared by other agoraphobics .


Hypnosis in palliative care: from clinical insights to the science of self-regulation.
Palliative care spans a wide-ranging spectrum: from pain-management to spiritual support. As the demand for end-of-life care increases, so does the demand for innovative, effective, interventions. Mind-body techniques seem especially advantageous in a palliative context. Here we show that hypnosis serves an excellent adjunct therapy in palliative care to boost the efficacy of standard treatments. With the overarching goal of bridging clinical and scientific insights, we outline how five core principles of hypnosis can benefit the diverse needs of palliative care.


Hair loss and hair-pulling in rhesus macaques (Macaca mulatta).
Alopecia is a common problem in rhesus macaque colonies. A possible cause of this condition is hair-pulling; however the true relationship between hair-pulling and alopecia is unknown. The purpose of this study was to examine the relationship between hair loss and hair-pulling in 1258 rhesus macaques housed in 4 primate colonies across the United States. Alopecia levels ranged from 34.3% to 86.5% (mean, 49.3%) at the primate facilities. At facilities reporting a sex-associated difference, more female macaques were reported to exhibit alopecia than were males. In contrast, more males were reported to hair-pull. Animals reported to hair-pull were significantly more likely to have some amount of alopecia, but rates of hair-pulling were substantially lower than rates of alopecia, ranging from 0.6% to 20.5% (mean, 7.7%) of the populations. These results further demonstrate that hair-pulling plays only a small role in alopecia in rhesus macaques.


Occurrence, distribution and bioaccumulation of antibiotics in the Haihe River in China.
The occurrence, distribution and bioaccumulation of 22 antibiotics, including eight fluoroquinolones (FQs), nine sulfonamides (SAs) and five macrolides (MLs), in the Haihe River were investigated. Surface water, sediment and fish samples were analyzed by HPLC-MS/MS. Ofloxacin, sulfamethoxazole, sulfadiazine, erythromycin and roxithromycin were found in all surface water samples. SAs were the predominant antibiotics in surface water samples in the mainstream of Haihe River, with the maximum concentration of 201 ng L(-1) for sulfamethoxazole. Compared with Haihe River, higher concentrations of antibiotics were found in the surface water samples in the Dagu and Chentaizi Drainage Rivers. The antibiotics pollution in the mainstream of Haihe River may derive from its tributaries, whereas the wastewater from fish ponds was not the main source of FQs, SAs and MLs in the river water. In sediment samples, FQs were the most frequently detected antibiotics and their concentrations were much higher than SAs and MLs. The high concentrations and detection frequencies of FQs in sediments indicate that sediments are an important reservoir for FQs as they are strongly adsorbed onto sediments. Ciprofloxacin and erythromycin exhibited potential bioaccumulation in crucian carp, with mean bioaccumulation factors of 3262 L kg(-1) and 4492 L kg(-1), respectively.


Effect of COVID-19 related lockdown on ophthalmic practice and patient care in India: Results of a survey.
In early 2020, the World Health Organization declared the outbreak of the disease COVID-19, caused by a new variant of coronavirus 2019-nCoV as a global pandemic. The government of India ordered a nationwide lockdown for 21 days, limiting movement of people as a preventive measure. This survey was designed and conducted during the lockdown period to assess its effect on ophthalmic practice and patient care in India. An online survey was sent across to practicing Indian ophthalmologists across through various social media platforms. All valid responses were tabulated and analyzed. A total of 1260 ophthalmologists responded to the survey. Most of the respondents (775/1260; 61.5%) were in private practice and 14.8% (187/1260) were affiliated to ophthalmic institutes. At the time of taking the survey, 72.5% of the respondents (913/1260) were not seeing any patients due to the lockdown. Of those who were still examining patients, 82.9% (287/347) were only seeing emergency cases, based on their own clinical judgement. The proportion of ophthalmologists in ophthalmic institutes, government and municipal hospitals (126/253;49.8%) who were still seeing patients was significantly higher (P < 0.0001) than those in private practice (174/775;22.4%). Apart from emergencies such as trauma, retinal detachment, and endophthalmitis (81.8%), other surgeries that were still being performed included intravitreal injections (9.1%) and cataract surgeries (5.9%). Approximately, 77.5% (976/1260) of the respondents had begun telephonic/e-mail/video consultations or consultations over social media applications since the lockdown began. In addition, 59.1% (745/1260) felt that ophthalmologists were potentially at a higher risk of contracting COVID-19 compared to other specialties while examining patients. When asked about the resumption of practice upon easing off of the restrictions, 57.8% (728/1260) of the respondents said they were unsure of when to resume elective surgeries; furthermore, 62.8% (791/1260) were unsure about the preferred screening strategy or precautionary approach prior to resuming surgeries and were awaiting guidelines. Our survey shows that majority of ophthalmologists in India were not seeing patients during the COVID-19 lockdown, with near-total cessation of elective surgeries. Emergency services were still being attended to by 27.5% of ophthalmologists who responded. A large proportions of ophthalmologists had switched over to telephonic advice or other forms of telemedicine to assist patients. Most of the responding ophthalmologists were unclear about when and how to resume surgeries upon easing off of the COVID-19 related restrictions. Regulatory bodies should take note of this and issue appropriate guidelines regarding the same.


Macronutrient Regulation of Ghrelin and Peptide YY in Pediatric Obesity and Prader-Willi Syndrome.
The roles of macronutrients and GH in the regulation of food intake in pediatric obesity and Prader-Willi Syndrome (PWS) are poorly understood. We compared effects of high-carbohydrate (HC) and high-fat (HF) meals and GH therapy on ghrelin, insulin, peptide YY (PYY), and insulin sensitivity in children with PWS and body mass index (BMI) -matched obese controls (OCs). In a randomized, crossover study, 14 PWS (median, 11.35 y; BMI z score [BMI-z], 2.15) and 14 OCs (median, 11.97 y; BMI-z, 2.35) received isocaloric breakfast meals (HC or HF) on separate days. Blood samples were drawn at baseline and every 30 minutes for 4 hours. Mixed linear models were adjusted for age, sex, and BMI-z. Relative to OCs, children with PWS had lower fasting insulin and higher fasting ghrelin and ghrelin/PYY. Ghrelin levels were higher in PWS across all postprandial time points (P < .0001). Carbohydrate was more potent than fat in suppressing ghrelin levels in PWS (P = .028); HC and HF were equipotent in OCs but less potent than in PWS (P = .011). The increase in PYY following HF was attenuated in PWS (P = .037); thus, postprandial ghrelin/PYY remained higher throughout. A lesser increase in insulin and lesser decrease in ghrelin were observed in GH-treated PWS patients than in untreated patients; PYY responses were comparable. Children with PWS have fasting and postprandial hyperghrelinemia and an attenuated PYY response to fat, yielding a high ghrelin/PYY ratio. GH therapy in PWS is associated with increased insulin sensitivity and lesser postprandial suppression of ghrelin. The ratio Ghrelin/PYY may be a novel marker of orexigenic drive.


Radiographic identification of the primary medial knee structures.
Radiographic landmarks for medial knee attachment sites during anatomic repairs or reconstructions are unknown. If identified, they could assist in the preoperative evaluation of structure location and allow for postoperative assessment of reconstruction tunnel placement. Radiopaque markers were implanted into the femoral and tibial attachments of the superficial medial collateral ligament and the femoral attachments of the posterior oblique and medial patellofemoral ligaments of eleven fresh-frozen, nonpaired cadaveric knee specimens. Both anteroposterior and lateral radiographs were made. Structures were assessed within quadrants formed by the intersection of reference lines projected on the lateral radiographs. Quantitative measurements were performed by three independent examiners. Intraobserver reproducibility and interobserver reliability were determined with use of intraclass correlation coefficients. The overall intraclass correlation coefficients for intraobserver reproducibility and interobserver reliability were 0.996 and 0.994, respectively. On the anteroposterior radiographs, the attachment sites of the superficial medial collateral ligament, posterior oblique ligament, and medial patellofemoral ligament were 30.5 +/- 2.4 mm, 34.8 +/- 2.7 mm, and 42.3 +/- 2.1 mm from the femoral joint line, respectively. On the lateral femoral radiographs, the attachment of the superficial medial collateral ligament was 6.0 +/- 0.8 mm from the medial epicondyle and was located in the anterodistal quadrant. The attachment of the posterior oblique ligament was 7.7 +/- 1.9 mm from the gastrocnemius tubercle and was located in the posterodistal quadrant. The attachment of the medial patellofemoral ligament was 8.9 +/- 2.0 mm from the adductor tubercle and was located in the anteroproximal quadrant. On the lateral tibial radiographs, the proximal and distal tibial attachments of the superficial medial collateral ligament were 15.9 +/- 5.2 and 66.1 +/- 3.6 mm distal to the tibial inclination, respectively. The attachment locations of the main medial knee structures can be qualitatively and quantitatively correlated to osseous landmarks and projected radiographic lines, with close agreement among examiners.


Noniterative interpolation-based super-resolution minimizing aliasing in the reconstructed image.
Super-resolution (SR) techniques produce a high-resolution image from a set of low-resolution undersampled images. In this paper, we propose a new method for super-resolution that uses sampling theory concepts to derive a noniterative SR algorithm. We first raise the issue of the validity of the data model usually assumed in SR, pointing out that it imposes a band-limited reconstructed image plus a certain type of noise. We propose a sampling theory framework with a prefiltering step that allows us to work with more general data models and also a specific new method for SR that uses Delaunay triangulation and B-splines to build the super-resolved image. The proposed method is noniterative and well posed. We prove its effectiveness against traditional iterative and noniterative SR methods on synthetic and real data. Additionally, we also prove that we can first solve the interpolation problem and then make the deblurring not only when the motion is translational but also when there are rotations and shifts and the imaging system Point Spread Function (PSF) is rotationally symmetric.


Ageing Increases Cardiac Electrical Remodelling in Rats and Mice via NOX4/ROS/CaMKII-Mediated Calcium Signalling.
Ageing is one of the risk factors associated with cardiovascular diseases including cardiac arrhythmias and heart failure. Ageing-related cardiac dysfunction involves a complicated pathophysiological progress. Abnormal membrane voltage and Ca2+ dynamics in aged cardiomyocytes contribute to ageing-related arrhythmias. However, its underlying mechanisms have not been well clarified. Young and old rats or mice were included in this study. Cardiac electrophysiological properties and functions were assessed by ECG, echocardiography, and ex vivo heart voltage and Ca2+ optical mapping. Proteomics, phosphor-proteomics, Western blotting, Masson staining, and ROS measurement were used to investigate the underlying mechanisms. Ageing increased the incidence of cardiac hypertrophy and fibrosis in rats. Moreover, ageing increased the occurrence of ventricular tachycardia or ventricular fibrillation induced by rapid pacing and during isoprenaline (ISO) (1 mg/kg i.p.) challenge in mice in vivo. Optical mapping with dual dyes (membrane voltage (V m ) dye and intracellular Ca2+ dye) simultaneously recording revealed that ageing increased the action potential duration (APD) and Ca2+ transient duration (CaTD) and slowed the ventricular conduction with the Langendorff-perfused mouse heart. More importantly, ageing increased the ISO-induced (1 μM) changes of APD (ΔAPD80) and CaTD (ΔCaTD50). Ageing also delayed the decay of Ca2+ transient by extending the decay time constant from 30% to 90% (τ 30-90). In addition, ageing decreased the V m /Ca 2+ latency which represented the coupling of V m /Ca 2+ including between the midpoint of AP depolarization and Ca2+ upstroke, peak transmembrane voltage and peak cytosolic calcium, and time to 50% voltage repolarization and extrusion of cytosolic calcium. Optical mapping also revealed that ageing increased the ISO-induced arrhythmia incidence and occurrence of the excitation rotor. Proteomics and phosphor-proteomics assays from rat hearts demonstrated ageing-induced protein and phosphor-protein changes, suggesting that CaMKII was involved in ageing-induced change. Ageing increased the level of ROS and the expression of NOX4, oxidative CaMKII (ox-CaMKII), phosphorated CaMKII (p-CaMKII), and periostin. Ageing accelerates cardiac remodelling and increases the susceptibility to ventricular arrhythmias through NOX4/ROS/CaMKII pathway-mediated abnormal membrane voltage and intracellular Ca2+ handling and V m /Ca 2+ coupling.


CD4(-)CD8alphaalpha subset of CD1d-restricted NKT cells controls T cell expansion.
Valpha24 invariant (Valpha24i) CD1d-restricted NKT cells are widely regarded to have immune regulatory properties. They are known to have a role in preventing autoimmune diseases and are involved in optimally mounted immune responses to pathogens and tumor cells. We were interested in understanding how these cells provide protection in autoimmune diseases. We first observed, using EBV/MHC I tetrameric complexes, that expansion of Ag-specific cells in human PBMCs was reduced when CD1d-restricted NKT cells were concomitantly activated. This was accompanied by an increase in a CD4(-)CD8alphaalpha(+) subset of Valpha24i NKT cells. To delineate if a specific subset of NKT cells was responsible for this effect, we generated different subsets of human CD4(-) and CD4(+) Valpha24i NKT clones and demonstrate that a CD4(-)CD8alphaalpha(+) subset with highly efficient cytolytic ability was unique among the clones in being able to suppress the proliferation and expansion of activated T cells in vitro. Activated clones were able to kill CD1d-bearing dendritic or target cells. We suggest that one mechanism by which CD1d-restricted NKT cells can exert a regulatory role is by containing the proliferation of activated T cells, possibly through timely lysis of APCs or activated T cells bearing CD1d.


Circulating HIV-specific CD8+ cytotoxic T cells express CD38 and HLA-DR antigens.
CD38, a molecule with multilineage distribution but unknown function, and the MHC class II molecule HLA-DR (DR) have markedly elevated levels of expression on CD8+ cells of HIV-infected people. This study investigated the expression of CD38 and DR Ag on circulating HIV-specific CD8+ CTL in HIV-seropositive subjects. Purified CD8+ lymphocytes from 22 participants in the University of California at Los Angeles Multicenter AIDS Cohort Study were screened for CTL activity against autologous EBV-immortalized lymphoblast targets infected with vaccinia vectors that carried HIVIIIB gag, pol, and env genes. Sixty-seven percent (14 of 21), 64% (14 of 22), and 9% (2 of 22), respectively, of the subjects had HIV-specific CD8+ CTL activity against gag, pol, and env proteins. CD8+ cells from 11 of the subjects who had high CTL activity were then FACS-separated using three-color immunofluorescence sorting. Circulating DR-CD38- CD8+ cells had little activity. Highly purified DR+CD38+ CD8+ cells had higher HIV-specific CTL activity than other CD8+ cells. DR+CD38- or DR-CD38+ CD8+ cells also mediated significant activity, but only about half as much on a per cell basis as DR+CD38+ CD8+ cells. This is the first report that the CD38 molecule is expressed in vivo on Ag-specific CD8+ CTL, and confirms previous reports that DR is expressed on these cells. Both asymptomatic HIV-seropositive subjects (144 +/- 132/mm3) and AIDS patients (253 +/- 178/mm3) had markedly elevated levels of DR+CD38+ CD8+ cells compared with the levels in HIV-seronegative controls (7 +/- 3/mm3). However, the level of anti-HIV CTL activity was not correlated with the level of DR+CD38+ CD8+ cells, indicating that enumeration of this lymphocyte population by flow cytometry most likely will not be a useful surrogate for measuring functional CTL activity. Low levels of HIV-specific CTL activity, especially against gag, were correlated with lower CD4+ cells numbers, suggesting that the loss of CD8+ T cell cytotoxic activity against HIV that has been reported to occur with advancing HIV disease progression may reflect in part the extent of CD4+ cell immunodeficiency in HIV-infected subjects.


Anacardic acid (6-pentadecylsalicylic acid) induces apoptosis of prostate cancer cells through inhibition of androgen receptor and activation of p53 signaling.
Anacardic acid (AA) is a mixture of 2-hydroxy-6-alkylbenzoic acid homologs. It is widely regarded as a non-specific histone acetyltransferase inhibitor of p300. The effects and the mechanisms of AA in LNCaP cells (prostate cancer cells) remain unknown. To investigate the effect of AA on LNCaP cells, we had carried out several experiments and found that AA inhibits LNCaP cell proliferation, induces G1/S cell cycle arrest and apoptosis of LNCaP cell. The mechanisms via which AA acts on LNCaP cells may be due to the following aspects. First, AA can regulate p300 transcription and protein level except for its mechanisms regulating function of p300 through post-translational modification in LNCaP cells. Second, AA can activate p53 through increasing the phosphorylation of p53 on Ser15 in LNCaP cells. AA can selectively activate p21 (target genes of p53). Third, AA can down-regulates androgen receptor (AR) through supressing p300. Our study suggests that AA has multiple anti-tumor activities in LNCaP cells and warrants further investigation.


Definition of the first mannosylation step in phosphatidylinositol mannoside synthesis. PimA is essential for growth of mycobacteria.
We examined the function of the pimA (Rv2610c) gene, located in the vicinity of the phosphatidylinositol synthase gene in the genomes of Mycobacterium tuberculosis and Mycobacterium smegmatis, which encodes a putative mannosyltransferase involved in the early steps of phosphatidylinositol mannoside synthesis. A cell-free assay was developed in which membranes from M. smegmatis overexpressing the pimA gene incorporate mannose from GDP-[(14)C]Man into di- and tri-acylated phosphatidylinositol mono-mannosides. Moreover, crude extracts from Escherichia coli producing a recombinant PimA protein synthesized diacylated phosphatidylinositol mono-mannoside from GDP-[(14)C]Man and bovine phosphatidylinositol. To determine whether PimA is an essential enzyme of mycobacteria, we constructed a pimA conditional mutant of M. smegmatis. The ability of this mutant to synthesize the PimA mannosyltransferase was dependent on the presence of a functional copy of the pimA gene carried on a temperature-sensitive rescue plasmid. We demonstrate here that the pimA mutant is unable to grow at the higher temperature at which the rescue plasmid is lost. Thus, the synthesis of phosphatidylinositol mono-mannosides and derived higher phosphatidylinositol mannosides in M. smegmatis appears to be dependent on PimA and essential for growth. This work provides the first direct evidence of the essentiality of phosphatidylinositol mannosides for the growth of mycobacteria.


Use of citrate adonitol agar as a selective medium for the isolation of Escherichia fergusonii from a captive reindeer herd.
Escherichia fergusonii is an emerging potentially zoonotic organism which has been recovered from a broad range of human and animal sources. Efforts to recover E. fergusonii from mixed flora hitherto however have been constrained by the lack of a suitable selective medium for its isolation. This paper reports for the first time the recovery of E. fergusonii from reindeer carcases in a wildlife park and the use of citrate adonitol agar to selectively screen for the presence of this organism in faecal samples from further animals in the park, and reindeer in their natural habitat in Norway.


Reliability and construct validity of the Dutch psychopathy checklist: youth version: findings from a sample of male adolescents in a juvenile justice treatment institution.
The present study examines the reliability and construct validity of the Dutch version of the Psychopathy Check List: Youth Version (PCL:YV) in a sample of male adolescents admitted to a secure juvenile justice treatment institution (N = 98). Hare's four-factor model is used to examine reliability and validity of the separate dimensions of psychopathy. Interrater reliabilities are good to excellent for the PCL:YV total score and most factor scores, except for the affective factor. Several suggestions are offered for optimizing reliability of this factor. Finally, meaningful associations between PCL:YV scores and scores on the Minnesota Multiphasic Personality Inventory-Adolescent and the Interpersonal Checklist-Revised support the construct validity of the PCL:YV total score as well as the four factors in the Dutch context.


Attitudes and Stereotypes in Lung Cancer versus Breast Cancer.
Societal perceptions may factor into the high rates of nontreatment in patients with lung cancer. To determine whether bias exists toward lung cancer, a study using the Implicit Association Test method of inferring subconscious attitudes and stereotypes from participant reaction times to visual cues was initiated. Participants were primarily recruited from an online survey panel based on US census data. Explicit attitudes regarding lung and breast cancer were derived from participants' ratings (n = 1778) regarding what they thought patients experienced in terms of guilt, shame, and hope (descriptive statements) and from participants' opinions regarding whether patients ought to experience such feelings (normative statements). Participants' responses to descriptive and normative statements about lung cancer were compared with responses to statements about breast cancer. Analyses of responses revealed that the participants were more likely to agree with negative descriptive and normative statements about lung cancer than breast cancer (P<0.001). Furthermore, participants had significantly stronger implicit negative associations with lung cancer compared with breast cancer; mean response times in the lung cancer/negative conditions were significantly shorter than in the lung cancer/positive conditions (P<0.001). Patients, caregivers, healthcare providers, and members of the general public had comparable levels of negative implicit attitudes toward lung cancer. These results show that lung cancer was stigmatized by patients, caregivers, healthcare professionals, and the general public. Further research is needed to investigate whether implicit and explicit attitudes and stereotypes affect patient care.


An inner cell wall protein (cwp1) from conidia of the entomopathogenic fungus Beauveria bassiana.
Following the removal of the rodlet layer from aerial or submerged conidia of the entomopathogenic deuteromycetous fungus Beauveria bassiana, SDS-insoluble, formic-acid-extractable proteins were found in the residual cell wall material. Two major proteins (12.8 and 14.0 kDa) were extracted with formic acid from fractured aerial and submerged conidia but not from blastospores. Oxidation of the sample extracted by formic acid resulted in a single protein band (15.4 kDa) as judged by SDS-PAGE. Antibodies against this cell wall protein (cwp1) did not cross-react with cell wall extracts from the entomopathogenic deuteromycetous fungi Verticillium lecanii or Metarhizium anisopliae. Western blot analysis of two-dimensional gels revealed at least three acidic isoforms (pI 4.0-4.8) of cwp1. Immunohistological studies revealed that the cwp1 was primarily localized in the cell wall of aerial and submerged conidia but not in blastospores. Immunolocalization was possible only if the conidia were previously boiled in 5% (v/v) beta-mercaptoethanol. The N-terminal sequence of cwp1 showed no similarities with other published sequences. Our results suggest that at least two major species of SDS-insoluble, formic-acid-extractable proteins exist in cell walls of B. bassiana aerial or submerged conidia; one is the hydrophobin which occurs in the outermost rodlet layer and the other, cwp1, occurs primarily next to the rodlet layer.


Problem-solving counseling for caregivers of the cognitively impaired: effective for whom?
Individualized problem-solving counseling for caregivers of cognitively impaired relatives is thought to help caregivers cope with the stress and burden of caregiving. Few studies have shown the effectiveness of counseling for these caregivers. To determine the effectiveness of individualized problem-solving counseling by nurses for caregivers and the expenditures of health care utilization. Caregivers (n = 77) of the cognitively impaired living at home were randomized to receive nurse counseling or not. Psychosocial adjustment to their relative's illness, psychological distress, burden, coping skills, and expenditures were measured after 6 months and 1 year. Although on average, all caregivers receiving nurse counseling indicated no improvement in psychosocial adjustment to their relative's illness, psychological distress, or caregiver burden, they found counseling very helpful and it was effective for a subgroup of caregivers. Those with poor logical analysis coping skills at baseline had decreased psychological distress (F(1,53) = 9.7, p = .003) and improved psychosocial adjustment (F(1,53) = 4.7, p = .035) after 1 year. Caregivers in control and counseling groups whose relatives entered a nursing home improved their psychosocial adjustment 23% on average whereas those continuing to live in the community decreased by 8%. Almost half as many relatives entered nursing homes in the counseling group (n = 9 vs. n = 5) but these compared to control group relatives had greater annualized per person expenditures for health and social services (Cdn$23,437 vs. Cdn$15,151). Caregivers found nurse counseling most helpful. Those indicating infrequent use of logical analysis coping skills showed benefits.


Evaluation of candidate reference genes for gene expression normalization in Brassica juncea using real time quantitative RT-PCR.
The real time quantitative reverse transcription PCR (qRT-PCR) is becoming increasingly important to gain insight into function of genes. Given the increased sensitivity, ease and reproducibility of qRT-PCR, the requirement of suitable reference genes for normalization has become important and stringent. It is now known that the expression of internal control genes in living organism vary considerably during developmental stages and under different experimental conditions. For economically important Brassica crops, only a couple of reference genes are reported till date. In this study, expression stability of 12 candidate reference genes including ACT2, ELFA, GAPDH, TUA, UBQ9 (traditional housekeeping genes), ACP, CAC, SNF, TIPS-41, TMD, TSB and ZNF (new candidate reference genes), in a diverse set of 49 tissue samples representing different developmental stages, stress and hormone treated conditions and cultivars of Brassica juncea has been validated. For the normalization of vegetative stages the ELFA, ACT2, CAC and TIPS-41 combination would be appropriate whereas TIPS-41 along with CAC would be suitable for normalization of reproductive stages. A combination of GAPDH, TUA, TIPS-41 and CAC were identified as the most suitable reference genes for total developmental stages. In various stress and hormone treated samples, UBQ9 and TIPS-41 had the most stable expression. Across five cultivars of B. juncea, the expression of CAC and TIPS-41 did not vary significantly and were identified as the most stably expressed reference genes. This study provides comprehensive information that the new reference genes selected herein performed better than the traditional housekeeping genes. The selection of most suitable reference genes depends on the experimental conditions, and is tissue and cultivar-specific. Further, to attain accuracy in the results more than one reference genes are necessary for normalization.


Fluorescence-Guided Probes of Aptamer-Targeted Gold Nanoparticles with Computed Tomography Imaging Accesses for in Vivo Tumor Resection.
Recent development of molecular imaging probes for fluorescence-guided surgery has shown great progresses for determining tumor margin to execute the tissue resection. Here we synthesize the fluorescent gold nanoparticles conjugated with diatrizoic acid and nucleolin-targeted AS1411 aptamer. The nanoparticle conjugates exhibit high water-solubility, good biocompatibility, visible fluorescence and strong X-ray attenuation for computed tomography (CT) contrast enhancement. The fluorescent nanoparticle conjugates are applied as a molecular contrast agent to reveal the tumor location in CL1-5 tumor-bearing mice by CT imaging. Furthermore, the orange-red fluorescence emitting from the conjugates in the CL1-5 tumor can be easily visualized by the naked eyes. After the resection, the IVIS measurements show that the fluorescence signal of the nanoparticle conjugates in the tumor is greatly enhanced in comparison to that in the controlled experiment. Our work has shown potential application of functionalized nanoparticles as a dual-function imaging agent in clinical fluorescence-guided surgery.


Ionizing radiation and inhibition of angiogenesis in a spontaneous mammary carcinoma and in a syngenic heterotopic allograft tumor model: a comparative study.
The combined treatment modality of ionizing radiation (IR) with inhibitors of angiogenesis (IoA) is a promising treatment modality based on preclinical in vivo studies using heterotopic xeno- and allograft tumor models. Nevertheless reservations still exist to translate this combined treatment modality into clinical trials, and more advanced, spontaneous orthotopic tumor models are required for validation to study the efficacy and safety of this treatment modality. We therefore investigated the combined treatment modality of IR in combination with the clinically relevant VEGF receptor (VEGFR) tyrosine kinase inhibitor PTK787 in the MMTV/c-neu induced mammary carcinoma model and a syngenic allograft tumor model using athymic nude mice. Mice were treated with fractionated IR, the VEGFR-inhibitor PTK787/ZK222584 (PTK787), or in combination, and efficacy and mechanistic-related endpoints were probed in both tumor models. Overall the treatment response to the IoA was comparable in both tumor models, demonstrating minimal tumor growth delay in response to PTK787 and PTK787-induced tumor hypoxia. Interestingly spontaneously growing tumors were more radiosensitive than the allograft tumors. More important combined treatment of irradiation with PTK787 resulted in a supraadditive tumor response in both tumor models with a comparable enhancement factor, namely 1.5 and 1.4 in the allograft and in the spontaneous tumor model, respectively. These results demonstrate that IR in combination with VEGF-receptor tyrosine kinase inhibitors is a valid, promising treatment modality, and that the treatment responses in spontaneous mammary carcinomas and syngenic allografts tumor models are comparable.


Adult mortality from leading sites of cancer in Italy and a note on European data.
An overview is given of recent patterns of trends in cancer mortality in various age groups in Italy and, for comparative purposes, in other European countries. Important changes in mortality for several major cancer sites have been observed over the last two decades, such as the declines in gastric cancer or the increases in lung cancer mortality. These are difficult to understand and justify from an epidemiological and public health viewpoint, since the main cause of lung cancer (cigarette smoking) has long been known, while the determinants of stomach cancer are still largely undefined. Changes were minor, and inconsistent across countries, for other major cancers, such as intestines or breast. Furthermore, trends were systematically more favourable in young adults (and, to a less extent middle age), as compared to the elderly. The decline in young adults reflects more favourable exposure to risk factors (eg declined smoking prevalence, better diet), but also improvements in diagnostic and therapeutic procedures for some cancers. Due caution is required in any extrapolation, particularly since the distribution of the most common types of cancer changes in subsequent age groups. These favourable trends in young adults offer, none the less, important (and positive) indications as to the likely future trends when the same generations will reach middle and older age.


LHX6, An Independent Prognostic Factor, Inhibits Lung Adenocarcinoma Progression through Transcriptional Silencing of β-catenin.
Introduction: Our previous study identified LIM homeobox domain 6 (LHX6) as a frequently epigenetically silenced tumor-suppressor gene in lung cancer. However, its clinical value has never been evaluated, and the in-depth anti-tumor mechanism remains unclear. Methods: Public database was used for lung cancer, lung adenocarcinoma and lung squamous carcinoma patients and tissue microarray data was used for lung adenocarcinoma patients to study prognostic outcome of LHX6 expression by Kaplan-Meier and Cox-regression analysis. In vitro proliferation, metastasis and in vivo nude mice model were used to evaluate the anti-tumor effect of LHX6 on lung adenocarcinoma cell lines. The mechanisms were explored using western blot, TOP/FOP flash assays and luciferase reporter assays. LHX6 expression and clinical stages data were collected from The Cancer Genome Atlas database (TCGA). Results: Expression of LHX6 was found to be a favorable independent prognostic factor for overall survival (OS) of total lung adenocarcinoma patients (P=0.014) and patients with negative lymph nodes status (P=0.014) but not related the prognostic outcome of lung squamous cell carcinoma patients. The expression status of LHX6 significantly correlated to histological grade (P<0.01), tumor size (P=0.026), lymph node status (P=0.039) and clinical stages (P<0.01) of lung adenocarcinoma patients. Functionally, LHX6 inhibited the proliferation and metastasis of lung adenocarcinoma cells in vitro and in vivo. Furthermore, LHX6 suppressed the Wnt/β-catenin pathway through transcriptionally silencing the expression of β-catenin, and the promoter region (-1161 bp to +27 bp) was crucial for its inhibitory activity. Conclusions: Our data indicate that the expression of LHX6 may serve as a favorable prognostic biomarker for lung adenocarcinoma patients and provide a novel mechanism of LHX6 involving in the tumorigenesis of lung adenocarcinoma.


Comparative sensitivity and specificity of the Casoni intradermal and the immunoelectrophoresis tests for the diagnosis of hydatid disease.
The Casoni intradermal (ID) test, using two antigens, was compared with the immunoelectrophoresis (IEP) test for diagnosis in 47 surgically confirmed cases of hydatid disease and 73 non-hydatid persons. An antigen prepared from boiled hydatid cyst fluid (HF) was markedly more sensitive in the ID test than another prepared from whole HF but both produced false-positive reactions in three persons with other disease conditions. The IEP test yielded no false positives and its sensitivity was similar to the ID test using the more sensitive boiled HF antigen. Some patients were ID-positive and IEP-negative and vice-versa. Diagnostic sensitivity of both tests varied according to the localization and condition of the cyst. A detectable immune response was more frequent in patients with liver cysts than in those with lung cysts. Regardless of cyst localization, lowest diagnostic sensitivity was observed in patients whose cysts were intact and of the hyaline type, whereas recently broken cysts were associated with the most consistently detectable immune response. The limitations of the ID test are discussed and it is suggested that, until more specific antigens are available, there appears to be little value in utilizing this test where the more specific serological techniques are available.


The role of harmonic expectancy violations in musical emotions: evidence from subjective, physiological, and neural responses.
The purpose of the present study was to investigate the effect of harmonic expectancy violations on emotions. Subjective response measures for tension and emotionality, as well as electrodermal activity (EDA) and heart rate (HR), were recorded from 24 subjects (12 musicians and 12 nonmusicians) to observe the effect of expectancy violations on subjective and physiological measures of emotions. In addition, an electro-encephalogram was recorded to observe the neural correlates for detecting these violations. Stimuli consisted of three matched versions of six Bach chorales, which differed only in terms of one chord (harmonically either expected, unexpected or very unexpected). Musicians' and nonmusicians' responses were also compared. Tension, overall subjective emotionality, and EDA increased with an increase in harmonic unexpectedness. Analysis of the event-related potentials revealed an early negativity (EN) for both the unexpected and the very unexpected harmonies, taken to reflect the detection of the unexpected event. The EN in response to very unexpected chords was significantly larger in amplitude than the EN in response to merely unexpected harmonic events. The ENs did not differ in amplitude between the two groups but peaked earlier for musicians than for nonmusicians. Both groups also showed a P3 component in response to the very unexpected harmonies, which was considerably larger for musicians and may reflect the processing of stylistic violations of Western classical music.


Ventricular arrhythmias during clinical treadmill testing and prognosis.
Although exercise-associated ventricular arrhythmias are frequently observed during exercise testing, their prognostic significance remains uncertain. Therefore, we aimed to evaluate the clinical correlates and prognostic significance of exercise-associated premature ventricular complexes (PVCs) during and after exercise testing. We studied 1847 heart failure-free patients who underwent clinical treadmill testing between March 13, 1997, and January 15, 2004, in the Veterans Affairs Palo Alto Health Care System. Logistic regression was used to evaluate the clinical and exercise test associations of exercise and recovery PVCs. Propensity score-adjusted Cox survival analyses were used to evaluate the prognostic significance of exercise-associated PVCs. Of the 1847 subjects, 850 (46.0%) developed exercise PVCs (median rate, 0.43 per minute) and 620 (33.6%) had recovery PVCs (median rate, 0.60 per minute). Resting PVCs, age, and systolic blood pressure were key predictors of both exercise and recovery PVCs. Whereas exercise PVCs were related to the heart rate increase with exercise, recovery PVCs were related to coronary disease (previous myocardial infarction, coronary revascularization procedure, or pathological Q waves on resting electrocardiogram) and ST-segment depression. During a 5.4-year mean follow-up, 161 subjects (8.7%) died, and 53 of these deaths (32.9%) were due to cardiovascular causes. Recovery PVCs, but not exercise PVCs, were associated with 71% to 96% greater propensity-adjusted mortality rates (hazard ratio, 1.96 [95% confidence interval, 1.31-2.91] for infrequent PVCs; hazard ratio, 1.71 [95% confidence interval, 1.07-2.73] for frequent PVCs compared with subjects without PVCs), and occurrence of recovery PVCs reclassified 33.2% of subjects with intermediate-risk Duke Treadmill Scores into higher-risk subgroups. In our heart failure-free population, recovery PVCs were associated with increased mortality and augmented established risk markers.


Thrombogenicity of hydrophilic and nonhydrophilic microcatheters and guiding catheters.
To assess in a swine model the in vivo thrombogenicity of various microcatheters and guiding catheters as a function of catheter material, catheter coating, and duration of implantation. Microcatheters (Tracker 18 and Fastracker 18, Target Therapeutics, Fremont, Calif; Magic 1.8, Balt, Montmorency, France; and Transit, Cordis Endovascular Systems, Miami Lakes, Fla) were placed through 6F guiding catheters (Fasguide, Target Therapeutics, and Envoy, Cordis Endovascular Systems) into the common carotid arteries of swine for 30 minutes (short term), 90 minutes (medium term), and 35 days (long term). Guiding catheters were implanted for 5 hours. At the end of the implantation periods the catheters were retracted and fixed for scanning electron microscopy. The surface of the Fastracker microcatheter was devoid of debris after both short- and medium-term implantation. The Tracker microcatheter had minimal accumulation of cellular elements whereas the Transit microcatheter showed moderate accumulation of nondeformed red blood cells. Neither the Tracker nor the Transit microcatheter showed evidence of increasing debris accumulation after medium-term implantation as compared with short-term implantation. The Magic microcatheter was coated with gross thrombus after both short- and medium-term implantation. The Fasguide guiding catheter was nearly devoid of debris, while the Envoy guiding catheter had moderate thrombus formation. Long-term implantation of the Fastracker microcatheter was well tolerated whereas that of the Transit catheter resulted in vessel occlusion. Hydrophilic microcatheters and guiding catheters are less thrombogenic than their nonhydrophilic counterparts, but not all hydrophilic coatings are equally hypothrombogenic. Degree of thrombogenicity depends on catheter material rather than surface morphology. Medium-term implantation did not yield increasing thrombus formation relative to short-term implantation.


Surface (S) Layer Proteins of Lactobacillus acidophilus Block Virus Infection via DC-SIGN Interaction.
Alphaviruses and flaviviruses are important human pathogens that include Chikungunya virus (CHIKV), Dengue virus (DENV), and Zika virus (ZIKV), which can cause diseases in humans ranging from arthralgia to hemorrhagic fevers and microcephaly. It was previously shown that treatment with surface layer (S-layer) protein, present on the bacterial cell-envelope of Lactobacillus acidophilus, is able to inhibit viral and bacterial infections by blocking the pathogen's interaction with DC-specific intercellular adhesion molecule 3-grabbing non-integrin (DC-SIGN), a trans-membrane protein that is a C-type calcium-dependent lectin. DC-SIGN is known to act as an attachment factor for several viruses including alphaviruses and flaviviruses. In the present study, we used alphaviruses as a model system to dissect the mechanism of S-layer inhibition. We first evaluated the protective effect of S-layer using 3T3 cells, either wild type or stably expressing DC-SIGN, and infecting with the alphaviruses Semliki Forest virus (SFV) and CHIKV and the flaviviruses ZIKV and DENV. DC-SIGN expression significantly enhanced infection by all four viruses. Treatment of the cells with S-layer prior to infection decreased infectivity of all viruses only in cells expressing DC-SIGN. In vitro ELISA experiments showed a direct interaction between S-layer and DC-SIGN; however, confocal microscopy and flow cytometry demonstrated that S-layer binding to the cells was independent of DC-SIGN expression. S-layer protein prevented SFV binding and internalization in DC-SIGN-expressing cells but had no effect on virus binding to DC-SIGN-negative cells. Inhibition of virus binding occurred in a time-dependent manner, with a significant reduction of infection requiring at least a 30-min pre-incubation of S-layer with DC-SIGN-expressing cells. These results suggest that S-layer has a different mechanism of action compared to mannan, a common DC-SIGN-binding compound that has an immediate effect in blocking viral infection. This difference could reflect slower kinetics of S-layer binding to the DC-SIGN present at the plasma membrane (PM). Alternatively, the S-layer/DC-SIGN interaction may trigger the activation of signaling pathways that are required for the inhibition of viral infection. Together our results add important information relevant to the potential use of L. acidophilus S-layer protein as an antiviral therapy.


Evaluation of 4-Amino 2-Anilinoquinazolines against Plasmodium and Other Apicomplexan Parasites In Vitro and in a P. falciparum Humanized NOD-scid IL2Rγnull Mouse Model of Malaria.
A series of 4-amino 2-anilinoquinazolines optimized for activity against the most lethal malaria parasite of humans, Plasmodium falciparum, was evaluated for activity against other human Plasmodium parasites and related apicomplexans that infect humans and animals. Four of the most promising compounds from the 4-amino 2-anilinoquinazoline series were equally as effective against the asexual blood stages of the zoonotic P. knowlesi, suggesting that they could also be effective against the closely related P. vivax, another important human pathogen. The 2-anilinoquinazoline compounds were also potent against an array of P. falciparum parasites resistant to clinically available antimalarial compounds, although slightly less so than against the drug-sensitive 3D7 parasite line. The apicomplexan parasites Toxoplasma gondii, Babesia bovis, and Cryptosporidium parvum were less sensitive to the 2-anilinoquinazoline series with a 50% effective concentration generally in the low micromolar range, suggesting that the yet to be discovered target of these compounds is absent or highly divergent in non-Plasmodium parasites. The 2-anilinoquinazoline compounds act as rapidly as chloroquine in vitro and when tested in rodents displayed a half-life that contributed to the compound's capacity to clear P. falciparum blood stages in a humanized mouse model. At a dose of 50 mg/kg of body weight, adverse effects to the humanized mice were noted, and evaluation against a panel of experimental high-risk off targets indicated some potential off-target activity. Further optimization of the 2-anilinoquinazoline antimalarial class will concentrate on improving in vivo efficacy and addressing adverse risk.


Flk2/Flt3 promotes both myeloid and lymphoid development by expanding non-self-renewing multipotent hematopoietic progenitor cells.
Defining differentiation pathways is central to understanding the pathogenesis of hematopoietic disorders, including leukemia. The function of the receptor tyrosine kinase Flk2 (Flt3) in promoting myeloid development remains poorly defined, despite being commonly mutated in acute myeloid leukemia. We investigated the effect of Flk2 deficiency on myelopoiesis, focusing on specification of progenitors between HSC and mature cells. We provide evidence that Flk2 is critical for proliferative expansion of multipotent progenitors that are common precursors for all lymphoid and myeloid lineages, including megakaryocyte/erythroid (MegE) cells. Flk2 deficiency impaired the generation of both lymphoid and myeloid progenitors by abrogating propagation of their common upstream precursor. At steady state, downstream compensatory mechanisms masked the effect of Flk2 deficiency on mature myeloid output, whereas transplantation of purified progenitors revealed impaired generation of all mature lineages. Flk2 deficiency did not affect lineage choice, thus dissociating the role of Flk2 in promoting cell expansion and regulating cell fate. Surprisingly, despite impairing myeloid development, Flk2 deficiency afforded protection against myeloablative insult. This survival advantage was attributed to reduced cell cycling and proliferation of progenitors in Flk2-deficient mice. Our data support the existence of a common Flk2(+) intermediate for all hematopoietic lineages and provide insight into how activating Flk2 mutations promote hematopoietic malignancy by non-Flk2-expressing myeloid cells.


Mechanisms of hearing loss after blast injury to the ear.
Given the frequent use of improvised explosive devices (IEDs) around the world, the study of traumatic blast injuries is of increasing interest. The ear is the most common organ affected by blast injury because it is the body's most sensitive pressure transducer. We fabricated a blast chamber to re-create blast profiles similar to that of IEDs and used it to develop a reproducible mouse model to study blast-induced hearing loss. The tympanic membrane was perforated in all mice after blast exposure and found to heal spontaneously. Micro-computed tomography demonstrated no evidence for middle ear or otic capsule injuries; however, the healed tympanic membrane was thickened. Auditory brainstem response and distortion product otoacoustic emission threshold shifts were found to be correlated with blast intensity. As well, these threshold shifts were larger than those found in control mice that underwent surgical perforation of their tympanic membranes, indicating cochlear trauma. Histological studies one week and three months after the blast demonstrated no disruption or damage to the intra-cochlear membranes. However, there was loss of outer hair cells (OHCs) within the basal turn of the cochlea and decreased spiral ganglion neurons (SGNs) and afferent nerve synapses. Using our mouse model that recapitulates human IED exposure, our results identify that the mechanisms underlying blast-induced hearing loss does not include gross membranous rupture as is commonly believed. Instead, there is both OHC and SGN loss that produce auditory dysfunction.


Endoscopic transcanal approach to the middle ear for management of pediatric cholesteatoma.
To describe outcomes for endoscopic ear surgery (EES) for pediatric cholesteatoma at a tertiary pediatric hospital. Retrospective case series of 65 pediatric cholesteatoma cases in 38 ears. Subgrouping based on cholesteatoma type and EES type. Surgical findings, outcomes, and demographic data were evaluated. Endoscopes were used in 65 pediatric cholesteatoma cases in 38 primary ears (34 patients), followed for an average of 2.6 years (9 months to 4.6 years). The endoscope was used as the primary visualization tool in 31 (81.6%) ears (EES 2 or 3), and as an adjunct to the microscope in seven ears (EES 1). Twenty-two (57.9%) ears and 35 (53.4%) cases were transcanal endoscopic ear surgery (EES 3 or TEES). Overall, there was recurrence in five (13.2%) ears and residual in four (10.5%) ears. Cholesteatoma was acquired in 27 ears, with average age 10.9 years; and congenital in 11 ears, with average age 3.8 years. Surgical time was longer for acquired cases (226 vs. 154 minutes). Hearing outcomes were comparable for both cholesteatoma types. Residual disease was seen in three (11.1%) acquired ears and one (9.1%) congenital ear. Overall, the lowest rates of recurrent and residual disease were seen in EES 3 cases, and relatively low rates in EES 2 and 3 ears, including four (12.9%) recurrences and two (6.5%) ears with residual disease. The endoscopes are a viable tool for resection of pediatric cholesteatoma and provide excellent visualization of the middle ear and associated recesses. 4. Laryngoscope, 127:2653-2658, 2017.


Transporting an ionic-liquid/water mixture in a conical nanochannel: a nanofluidic memristor.
Synthetic conical nanochannels have gained attention for their ion rectification behavior, which can be used to mimic the functionalities of biological ion channels. Employing these nanochannels and inspired by biological synaptic dynamics, we herein demonstrate a new nanofluidic device as a memristor, the ion transport conductance of which depends on the history of its ion flow. The nanofluidic memristors show excellent repeatability, high ON/OFF ratios, and sufficiently long retention times, which are highly desirable for logic control and neuromorphic engineering applications in nanofluidic systems as well as for fundamental transport studies of ionic liquids on a nanoscale.


QTL × environment interactions underlie ionome divergence in switchgrass.
Ionomics measures elemental concentrations in biological organisms and provides a snapshot of physiology under different conditions. In this study, we evaluate genetic variation of the ionome in outbred, perennial switchgrass in three environments across the species' native range, and explore patterns of genotype-by-environment interactions. We grew 725 clonally replicated genotypes of a large full sib family from a four-way linkage mapping population, created from deeply diverged upland and lowland switchgrass ecotypes, at three common gardens. Concentrations of 18 mineral elements were determined in whole post-anthesis tillers using ion coupled plasma mass spectrometry (ICP-MS). These measurements were used to identify quantitative trait loci (QTL) with and without QTL-by-environment interactions (QTLxE) using a multi-environment QTL mapping approach. We found that element concentrations varied significantly both within and between switchgrass ecotypes, and GxE was present at both the trait and QTL level. Concentrations of 14 of the 18 elements were under some genetic control, and 77 QTL were detected for these elements. Seventy-four percent of QTL colocalized multiple elements, half of QTL exhibited significant QTLxE, and roughly equal numbers of QTL had significant differences in magnitude and sign of their effects across environments. The switchgrass ionome is under moderate genetic control and by loci with highly variable effects across environments.


Conversion therapy for homosexuality: serious violation of ethics.
Across the world, homosexuality is gaining legitimacy; stigma and discrimination are gradually giving way to equality and inclusion. The situation in India is in stark contrast to these trends. In this country, homosexuality is an offence as per Section 377 of the IPC. The homosexual community is fighting for its rights and continues to suffer from intense stigma and discrimination. Their healthcare needs are not at all attended to; their sexual orientation is conceptualised as a socially deviant mental disease that needs psychiatric treatment.


Clinical intestinal transplantation: new perspectives and immunologic considerations.
Although tacrolimus-based immunosuppression has made intestinal transplantation feasible, the risk of the requisite chronic high-dose treatment has inhibited the widespread use of these procedures. We have examined our 1990-1997 experience to determine whether immunomodulatory strategies to improve outlook could be added to drug treatment. Ninety-eight consecutive patients (59 children, 39 adults) with a panoply of indications received 104 allografts under tacrolimus-based immunosuppression: intestine only (n = 37); liver and intestine (n = 50); or multivisceral (n = 17). Of the last 42 patients, 20 received unmodified adjunct donor bone marrow cells; the other 22 were contemporaneous control patients. With a mean followup of 32 +/- 26 months (range, 1-86 months), 12 recipients (3 intestine only, 9 composite grafts) are alive with good nutrition beyond the 5-year milestone. Forty-seven (48%) of the total group survive bearing grafts that provide full (91%) or partial (9%) nutrition. Actuarial patient survival at 1 and 5 years (72% and 48%, respectively) was similar with isolated intestinal and composite graft recipients, but the loss rate of grafts from rejection was highest with intestine alone. The best results were in patients between 2 and 18 years of age (68% at 5 years). Adjunct bone marrow did not significantly affect the incidence of graft rejection, B-cell lymphoma, or the rate or severity of graft-versus-host disease. These results demonstrate that longterm rehabilitation similar to that with the other kinds of organ allografts is achievable with all three kinds of intestinal transplant procedures, that the morbidity and mortality is still too high for their widespread application, and that the liver is significantly but marginally protective of concomitantly engrafted intestine. Although none of the endpoints were markedly altered by donor leukocyte augmentation (and chimerism) with bone marrow, establishment of the safety of this adjunct procedure opens the way to further immune modulation strategies that can be added to the augmentation protocol.


Discovery of a P450-catalyzed step in vindoline biosynthesis: a link between the aspidosperma and eburnamine alkaloids.
Here we report the discovery of a cytochrome P450 that is required for the biosynthesis of vindoline, a plant-derived natural product used for semi-synthesis of several anti-cancer drugs. This enzyme catalyzes the formation of an epoxide that can undergo rearrangement to yield the vincamine-eburnamine backbone, thereby providing evidence for the long-standing hypothesis that the aspidosperma- and eburnamine-type alkaloids are biosynthetically related.


Comparison of Kriging and Moving Least Square Methods to Change the Geometry of Human Body Models.
Finite Element Human Body Models (HBM) have become powerful tools to study the response to impact. However, they are typically only developed for a limited number of sizes and ages. Various approaches driven by control points have been reported in the literature for the non-linear scaling of these HBM into models with different geometrical characteristics. The purpose of this study is to compare the performances of commonly used control points based interpolation methods in different usage scenarios. Performance metrics include the respect of target, the mesh quality and the runability. For this study, the Kriging and Moving Least square interpolation approaches were compared in three test cases. The first two cases correspond to changes of anthropometric dimensions of (1) a child model (from 6 to 1.5 years old) and (2) the GHBMC M50 model (Global Human Body Models Consortium, from 50th to 5th percentile female). For the third case, the GHBMC M50 ribcage was scaled to match the rib cage geometry derived from a CT-scan. In the first two test cases, all tested methods provided similar shapes with acceptable results in terms of time needed for the deformation (a few minutes at most), overall respect of the targets, element quality distribution and time step for explicit simulation. The personalization of rib cage proved to be much more challenging. None of the methods tested provided fully satisfactory results at the level of the rib trajectory and section. There were corrugated local deformations unless using a smooth regression through relaxation. Overall, the results highlight the importance of the target definition over the interpolation method.


Autofluorescence spectroscopy for in vivo diagnosis of DMBA-induced hamster buccal pouch pre-cancers and cancers.
Our previous ex vivo study has shown that autofluorescence spectroscopy at 330-nm excitation can discriminate specimens of normal buccal pouch mucosa (normal), epithelial hyperkeratosis (hyperkeratosis), epithelial dysplasia (dysplasia), and squamous cell carcinoma (SCC) taken from DMBA-treated hamsters by using the method of partial least-squares discriminant analysis (PLSDA). This study used a fiber optics-based fluorescence spectroscopy system to measure the autofluorescence spectra of 23 normal, 14 hyperkeratosis, 28 dysplasia, and 10 SCC samples in vivo. PLSDA with cross-validation was used to analyze the autofluorescence spectral data of all samples. We found that at 330-nm excitation, the autofluorescence spectra of all samples had two main peaks: one at 380 nm and the other at 460 nm. The hyperkeratosis samples had a higher 380-nm emission peak (EP) and a lower 460-nm EP than normal samples. On the contrary, the dysplasia samples had a lower 380-nm EP and a higher 460-nm EP than normal samples. Furthermore, the SCC samples had a much lower 380-nm EP and a much higher 460-nm EP than all other samples. To quantify the spectral changes during the progression of oral carcinogenesis, ratios of the area under the spectrum of 380 +/- 15 nm to that under the spectrum of 460 +/- 15 nm (denoted as A(380 +/- 15 nm)/A(460 +/- 15 nm)) for all samples were calculated. The mean ratio values of A(380 +/- 15 nm)/A(460 +/- 15 nm) decreased gradually from hyperkeratosis to normal, to dysplasia, and to SCC samples. Significant differences in this mean ratio were found between any two groups of normal, hyperkeratosis, dysplasia, and SCC samples. By choosing proper thresholds, PLSDA with cross-validation could provide an accurate identification rate of 86% for hyperkeratosis, of 87% for normal, and of 89% for dysplasia samples. In addition, by choosing a proper threshold, we could separate benign (normal and hyperkeratosis) from dysplasia or SCC tissues with a sensitivity of 92% and a specificity of 95%. Our results indicate that the autofluorescence spectroscopy technique is a useful diagnostic tool for in vivo diagnosis of oral pre-cancers and cancers in DMBA-induced hamster buccal pouch carcinogenesis model.


Exposures to phthalates and bisphenols in pregnancy and postpartum weight gain in a population-based longitudinal birth cohort.
Experimental evidence suggests that exposures to phthalates and bisphenols may interfere with processes related to glucose and lipid metabolism, insulin sensitivity, and body weight. Few studies have considered the possible influence of chemical exposures during pregnancy on maternal weight gain or metabolic health outcomes postpartum. To examine the associations of early and mid-pregnancy bisphenol and phthalate urine concentrations with maternal weight gain 6 years postpartum. We analyzed urine samples for bisphenol, phthalate and creatinine concentrations from early and mid-pregnancy in 1192 women in a large, population-based birth cohort in Rotterdam, the Netherlands, and examined postpartum weight gain using maternal anthropometrics before pregnancy and 6 years postpartum. We have used covariate-adjusted linear regressions to evaluate associations of early and mid-pregnancy bisphenols and phthalate metabolites with weight change. Mediator and interaction models have been used to assess the role of gestational weight gain and breastfeeding, respectively. Sensitivity analysis is performed among women without subsequent pregnancies. Among all 1192 mothers included in the analysis, each log unit increase in the average bisphenol A and all assessed phthalate groupings were associated with increased maternal weight gain. As a proxy for phthalate exposure, each log unit increase in averaged phthalic acid was associated with 734 g weight gain (95% CI 273-1196 g) between pre-pregnancy and 6 years postpartum. Mediation by gestational weight gain was not present. Breastfeeding and ethnicity did not modify the effects. Stratification revealed these associations to be strongest among overweight and obese women. Among women without subsequent pregnancies (n = 373) associations of bisphenols, HMW phthalate metabolites and di-2-ethylhexylphthalate metabolites attenuated. For phthalic acid, LMW phthalate metabolites and di-n-octylphthalate metabolites associations increased. Similarly to the whole group, stratification yielded significant results among overweight and obese women. In a large population-based birth cohort, early and mid-pregnancy phthalate exposures are associated with weight gain 6 years postpartum, particularly among overweight and obese women. These data support ongoing action to replace phthalates with safer alternatives.


Nitrogen Use Efficiency in Sorghum: Exploring Native Variability for Traits Under Variable N-Regimes.
Exploring the natural genetic variability and its exploitation for improved Nitrogen Use Efficiency (NUE) in sorghum is one of the primary goals in the modern crop improvement programs. The integrated strategies include high-throughput phenotyping, next generation sequencing (NGS)-based genotyping technologies, and a priori selected candidate gene studies that help understand the detailed physiological and molecular mechanisms underpinning this complex trait. A set of sixty diverse sorghum genotypes was evaluated for different vegetative, reproductive, and yield traits related to NUE in the field (under three N regimes) for two seasons. Significant variations for different yield and related traits under 0 and 50% N confirmed the availability of native genetic variability in sorghum under low N regimes. Sorghum genotypes with distinct genetic background had interestingly similar NUE associated traits. The Genotyping-By-Sequencing based SNPs (>89 K) were used to study the population structure, and phylogenetic groupings identified three distinct groups. The information of grain N and stalk N content of the individuals covered on the phylogenetic groups indicated randomness in the distribution for adaptation under variable N regimes. This study identified promising sorghum genotypes with consistent performance under varying environments, with buffer capacity for yield under low N conditions. We also report better performing genotypes for varied production use-grain, stover, and dual-purpose sorghum having differential adaptation response to NUE traits. Expression profiling of NUE associated genes in shoot and root tissues of contrasting lines (PVK801 and HDW703) grown in varying N conditions revealed interesting outcomes. Root tissues of contrasting lines exhibited differential expression profiles for transporter genes [ammonium transporter (SbAMT), nitrate transporters (SbNRT)]; primary assimilatory (glutamine synthetase (SbGS), glutamate synthase (SbGOGAT[NADH], SbGOGAT[Fd]), assimilatory genes [nitrite reductase (SbNiR[NADH]3)]; and amino acid biosynthesis associated gene [glutamate dehydrogenase (SbGDH)]. Identification and expression profiling of contrasting sorghum genotypes in varying N dosages will provide new information to understand the response of NUE genes toward adaptation to the differential N regimes in sorghum. High NUE genotypes identified from this study could be potential candidates for in-depth molecular analysis and contribute toward the development of N efficient sorghum cultivars.


Autism spectrum disorder.
Autism spectrum disorder is a construct used to describe individuals with a specific combination of impairments in social communication and repetitive behaviours, highly restricted interests and/or sensory behaviours beginning early in life. The worldwide prevalence of autism is just under 1%, but estimates are higher in high-income countries. Although gross brain pathology is not characteristic of autism, subtle anatomical and functional differences have been observed in post-mortem, neuroimaging and electrophysiological studies. Initially, it was hoped that accurate measurement of behavioural phenotypes would lead to specific genetic subtypes, but genetic findings have mainly applied to heterogeneous groups that are not specific to autism. Psychosocial interventions in children can improve specific behaviours, such as joint attention, language and social engagement, that may affect further development and could reduce symptom severity. However, further research is necessary to identify the long-term needs of people with autism, and treatments and the mechanisms behind them that could result in improved independence and quality of life over time. Families are often the major source of support for people with autism throughout much of life and need to be considered, along with the perspectives of autistic individuals, in both research and practice.