Factors influencing the infant feeding decision for socioeconomically deprived pregnant teenagers: the moral dimension.
The importance of breastfeeding-related health outcomes in reducing inequalities in health has been recognized as a National Health Service target to increase initiation rates especially among disadvantaged groups in England. This study examined the psychosocial factors influencing infant feeding intention among pregnant teenagers expecting their first baby and living in deprived urban areas in England. A mixed methods study, using a quantitative questionnaire based on the Theory of Planned Behaviour, was conducted in four English regions with predominantly white and Asian teenagers (n = 71). This method identified which of all known Theory of Planned Behaviour variables were the most important in influencing feeding intentions. Focus groups provided contextual insight into the meaning of these variables for white pregnant teenagers living in a northern English inner city (n = 17). Moral norms were identified as the most predictive variable influencing teenage intention to formula feed or breastfeed. The likelihood that breastfeeding "will be embarrassing" was the only attitudinal belief rated as significantly important in influencing teenage intention to breastfeed. Three overarching themes emerged from the focus group data: "moral norms,""sexuality of the breast," and "self-esteem," with concerns relating to breastfeeding in public cutting across all themes. Breastfeeding was viewed as a morally inappropriate behavior by most of these teenagers, with formula feeding being perceived as the appropriate behavior. Existing breastfeeding promotion activities are likely to continue to fail to reach teenagers experiencing deprivation in England in the absence of effective strategies to change the underlying negative moral norms toward breastfeeding.


Allele and haplotype frequency distribution in PTPN22 gene across variable ethnic groups: Implications for genetic association studies for autoimmune diseases.
The rs2476601-T allele at the protein tyrosine phosphatase non-receptor type 22 (PTPN22) gene has been consistently associated with several autoimmune diseases in European-derived populations. However, little is known about the allele and haplotype frequency distributions in PTPN22 among populations derived from other ethnic groups. In the present study, the allele and haplotype frequency distributions of six single nucleotide polymorphisms (SNPs) in the PTPN22 gene were compared among Brazilian populations and HapMap phase 3 dataset. A total of 10 different population samples were evaluated. Additionally, in admixed populations, individual genetic ancestries were estimated for Native American, African, and European contributions. Estimated individual ancestries were used as quantitative traits in a conditional approach for single-marker and haplotype-specific regression analyses. It was shown that several SNPs and haplotypes have different frequencies among different ethnic populations. Individual genetic ancestries were not associated with the rs2476601-T allele, but were associated with PTPN22 haplotypes in Brazilian, Mexican, and African-American admixed populations. Our results suggest caution in the interpretation of results found in association studies involving PTPN22 polymorphisms in admixed populations. Correction for stratification generated by admixture should be mandatory to minimize or avoid chances of spurious association.


Comparison of between-training-sessions recovery strategies for world-class BMX pilots.
To assess the impact of between-training-sessions recovery strategies (passive [PAS], active [ACT], cold-water immersion [CWI], and ingestion of a recovery drink [NUTR]) on maximal cycling performance, perceptions of delayed-onset muscle soreness (DOMS), and fatigue in world-class BMX riders. Eleven elite BMX athletes, members of the French national team (top country in the 2011 international ranking, 4 medals at the 2012 World Championships, top European country), participated in the study, which involved standardized training periods. Athletes performed 3 maximal-sprint power tests: the first day of the week before the training session and before and after training on the third day of the week (D3). The recovery strategy was randomly assigned to each participant on day 2 immediately after the last training period of the day. Perceptions of DOMS and general fatigue were recorded on D3. After training on D3, the decrease in maximal-sprint power (Pmax) was significantly greater for PAS than with CWI (P=.02) and NUTR (P=.018). Similar results were found with ACT (vs CWI P=.044, and vs NUTR P=.042). Self-reported DOMS and fatigue were significantly greater after PAS than after other strategies. For elite BMX riders, between training days, nutritional and/or CWI recovery strategies appear to be best for reducing muscle fatigue and increasing the capacity to withstand the training schedule.


Choice of Adaptive Sampling Strategy Impacts State Discovery, Transition Probabilities, and the Apparent Mechanism of Conformational Changes.
Interest in atomically detailed simulations has grown significantly with recent advances in computational hardware and Markov state modeling (MSM) methods, yet outstanding questions remain that hinder their widespread adoption. Namely, how do alternative sampling strategies explore conformational space and how might this influence predictions generated from the data? Here, we seek to answer these questions for four commonly used sampling methods: (1) a single long simulation, (2) many short simulations run in parallel, (3) adaptive sampling, and (4) our recently developed goal-oriented sampling algorithm, FAST. We first develop a theoretical framework for analytically calculating the probability of discovering select states on simple landscapes, where we uncover the drastic effects of varying the number and length of simulations. We then use kinetic Monte Carlo simulations on a variety of physically inspired landscapes to characterize the probability of discovering particular states and transition pathways for each of the four methods. Consistently, we find that FAST simulations discover each target state with the highest probability, while traversing realistic pathways. Furthermore, we uncover the potential pathology that short parallel simulations sometimes predict an incorrect transition pathway by crossing large energy barriers that long simulations would typically circumnavigate. We refer to this pathology as "pathway tunneling". To protect against this phenomenon when using adaptive-sampling and FAST simulations, we introduce the FAST-string method. This method enhances sampling along the highest-flux transition paths to refine an MSMs transition probabilities and discriminate between competing pathways. Additionally, we compare the performance of a variety of MSM estimators in describing accurate thermodynamics and kinetics. For adaptive sampling, we recommend simply normalizing the transition counts out of each state after adding small pseudocounts to avoid creating sources or sinks. Lastly, we evaluate whether our insights from simple landscapes hold for all-atom molecular dynamics simulations of the folding of the λ-repressor protein. Remarkably, we find that FAST-contacts predicts the same folding pathway as a set of long simulations but with orders of magnitude less simulation time.


The cross-cultural equivalence of participation instruments: a systematic review.
Concepts such as health-related quality of life, disability and participation may differ across cultures. Consequently, when assessing such a concept using a measure developed elsewhere, it is important to test its cultural equivalence. Previous research suggested a lack of cultural equivalence testing in several areas of measurement. This paper reviews the process of cross-cultural equivalence testing of instruments to measure participation in society. An existing cultural equivalence framework was adapted and used to assess participation instruments on five categories of equivalence: conceptual, item, semantic, measurement and operational equivalence. For each category, several aspects were rated, resulting in an overall category rating of 'minimal/none', 'partial' or 'extensive'. The best possible overall study rating was five 'extensive' ratings. Articles were included if the instruments focussed explicitly on measuring 'participation' and were theoretically grounded in the ICIDH(-2) or ICF. Cross-validation articles were only included if it concerned an adaptation of an instrument developed in a high or middle-income country to a low-income country or vice versa. Eight cross-cultural validation studies were included in which five participation instruments were tested (Impact on Participation and Autonomy, London Handicap Scale, Perceived Impact and Problem Profile, Craig Handicap Assessment Reporting Technique, Participation Scale). Of these eight studies, only three received at least two 'extensive' ratings for the different categories of equivalence. The majority of the cultural equivalence ratings given were 'partial' and 'minimal/none'. The majority of the 'none/minimal' ratings were given for item and measurement equivalence. The cross-cultural equivalence testing of the participation instruments included leaves much to be desired. A detailed checklist is proposed for designing a cross-validation study. Once a study has been conducted, the checklist can be used to ensure comprehensive reporting of the validation (equivalence) testing process and its results. • Participation instruments are often used in a different cultural setting than initial developed for. • The conceptualization of participation may vary across cultures. Therefore, cultural equivalence – the extent to which an instrument is equally suitable for use in two or more cultures – is an important concept to address. • This review showed that the process of cultural equivalence testing of the included participation instruments was often addressed insufficiently. • Clinicians should be aware that application of participations instruments in a different culture than initially developed for needs prior testing of cultural validity in the next context.


Early 1900 s detection of Batrachochytrium dendrobatidis in Korean amphibians.
The pathogenic fungus Batrachochytrium dendrobatidis (Bd) is a major conservation concern because of its role in decimating amphibian populations worldwide. We used quantitative PCR to screen 244 museum specimens from the Korean Peninsula, collected between 1911 and 2004, for the presence of Bd to gain insight into its history in Asia. Three specimens of Rugosa emeljanovi (previously Rana or Glandirana rugosa), collected in 1911 from Wonsan, North Korea, tested positive for Bd. Histology of these positive specimens revealed mild hyperkeratosis - a non-specific host response commonly found in Bd-infected frogs - but no Bd zoospores or zoosporangia. Our results indicate that Bd was present in Korea more than 100 years ago, consistent with hypotheses suggesting that Korean amphibians may be infected by endemic Asian Bd strains.


The draft genome assembly of Rhododendron delavayi Franch. var. delavayi.
Rhododendron delavayi Franch. is globally famous as an ornamental plant. Its distribution in southwest China covers several different habitats and environments. However, not much research had been conducted on Rhododendron spp. at the molecular level, which hinders understanding of its evolution, speciation, and synthesis of secondary metabolites, as well as its wide adaptability to different environments. Here, we report the genome assembly and gene annotation of R. delavayi var. delavayi (the second genome sequenced in the Ericaceae), which will facilitate the study of the family. The genome assembly will have further applications in genome-assisted cultivar breeding. The final size of the assembled R. delavayi var. delavayi genome (695.09 Mb) was close to the 697.94 Mb, estimated by k-mer analysis. A total of 336.83 gigabases (Gb) of raw Illumina HiSeq 2000 reads were generated from 9 libraries (with insert sizes ranging from 170 bp to 40 kb), achieving a raw sequencing depth of ×482.6. After quality filtering, 246.06 Gb of clean reads were obtained, giving ×352.55 coverage depth. Assembly using Platanus gave a total scaffold length of 695.09 Mb, with a contig N50 of 61.8 kb and a scaffold N50 of 637.83 kb. Gene prediction resulted in the annotation of 32 938 protein-coding genes. The genome completeness was evaluated by CEGMA and BUSCO and reached 95.97% and 92.8%, respectively. The gene annotation completeness was also evaluated by CEGMA and BUSCO and reached 97.01% and 87.4%, respectively. Genome annotation revealed that 51.77% of the R. delavayi genome is composed of transposable elements, and 37.48% of long terminal repeat elements (LTRs). The de novo assembled genome of R. delavayi var. delavayi (hereinafter referred to as R. delavayi) is the second genomic resource of the family Ericaceae and will provide a valuable resource for research on future comparative genomic studies in Rhododendron species. The availability of the R. delavayi genome sequence will hopefully provide a tool for scientists to tackle open questions regarding molecular mechanisms underlying environmental interactions in the genus Rhododendron, more accurately understand the evolutionary processes and systematics of the genus, facilitate the identification of genes encoding pharmaceutically important compounds, and accelerate molecular breeding to release elite varieties.


Neonatal alcohol exposure reduces NMDA induced Ca2+ signaling in developing cerebellar granule neurons.
Glutamatergic neurotransmission through NMDA receptors is critical for both neurogenesis and mature function of the central nervous system (CNS), and is thought to be one target for developmentally-induced damage by alcohol to brain function. In the current study we examined Ca2+ signaling linked to NMDA receptor activation as a potential site for alcohol's detrimental effects on the developing nervous system. We compared Ca2+ signals to NMDA in granule neurons cultured from cerebella of rat neonates exposed to alcohol (ethanol) during development with responses to NMDA recorded in separated control groups. Alcohol exposure was by the vapor chamber method on postnatal days 4-7. An intermittent exposure paradigm was used where the pups were exposed to alcohol vapor for 2. 5 h/day to produce peak BALs of approximately 320 mg%. Control pups were placed in an alcohol-free chamber for a similar time period or remained with their mother. After culture under alcohol-free conditions for up to 9 days, Ca2+ signaling in response to NMDA was measured using fura-2 Ca2+ imaging. Results show that the peak amplitude of the Ca2+ signal to NMDA was significantly smaller in cultured granule neurons obtained from alcohol-treated pups compared to granule neurons from control pups. In contrast, the Ca2+ signal to K+ depolarization was not depressed by the alcohol treatment. Resting Ca2+ levels were also altered by the alcohol treatment. These results show that intermittent alcohol exposure during development in vivo can induce long-term changes in CNS neurons that affect the Ca2+ signaling pathway linked to NMDA receptors and resting Ca2+ levels. Such changes could play an important role in the CNS dysfunction associated with alcohol exposure during CNS development.


Lyme carditis: cardiac abnormalities of Lyme disease.
We studied 20 patients, mostly young adult men, with cardiac involvement of Lyme disease. The commonest abnormality (18 patients) was fluctuating degrees of atrioventricular block; eight of them developed complete heart block. Thirteen patients had evidence of more diffuse cardiac involvement: electrocardiographic changes compatible with acute myopericarditis (11 patients), radionuclide evidence of mild left ventricular dysfunction (five of 12 patients tested), or frank cardiomegaly (one patient). Heart involvement was usually preceded by erythema chronicum migrans and sometimes accompanied by meningoencephalitis, facial palsy, arthritis, elevated serum IgM levels, or cryoglobulins containing IgM. The duration of cardiac involvement was usually brief (3 days to 6 weeks). The clinical picture in these patients has similarities to acute rheumatic fever; but in Lyme disease, complete heart block may be commoner, myopericardial involvement tends to be milder, and valves seem not to be affected.


Interaction of Object Binding Cues in Binaural Masking Pattern Experiments.
Object binding cues such as binaural and across-frequency modulation cues are likely to be used by the auditory system to separate sounds from different sources in complex auditory scenes. The present study investigates the interaction of these cues in a binaural masking pattern paradigm where a sinusoidal target is masked by a narrowband noise. It was hypothesised that beating between signal and masker may contribute to signal detection when signal and masker do not spectrally overlap but that this cue could not be used in combination with interaural cues. To test this hypothesis an additional sinusoidal interferer was added to the noise masker with a lower frequency than the noise whereas the target had a higher frequency than the noise. Thresholds increase when the interferer is added. This effect is largest when the spectral interferer-masker and masker-target distances are equal. The result supports the hypothesis that modulation cues contribute to signal detection in the classical masking paradigm and that these are analysed with modulation bandpass filters. A monaural model including an across-frequency modulation process is presented that account for this effect. Interestingly, the interferer also affects dichotic thresholds indicating that modulation cues also play a role in binaural processing.


Communication and compliance in eye casualty.
This study aimed to assess patients' perceptions and priorities when consulting doctors in eye casualty, to assess their satisfaction with eye casualty and to evaluate and improve patients' level of knowledge and understanding of their treatment. A selected consecutive group of 130 patients presenting to eye casualty between 1 July and 15 September 1995 was interviewed by two of the authors prior to collecting their medication. A further group was interviewed again after collecting their medication from the hospital pharmacist. The hospital pharmacist reiterated treatment details when patients collected their medication. Interviews were conducted by means of a questionnaire. There was no inter- or intraobserver variation. Patients' priorities and perceptions were measured as percentages of the group. Patient satisfaction was measured both by a score on a standardised questionnaire and on a visual analogue scale. Patient recall of treatment details was scored as correct or incorrect. The score prior to and after seeing the pharmacist was compared in those patients who were interviewed after collecting their medication. Among the patients 30.8% considered themselves emergencies, 20.8% were referred and the remainder attended for non-urgent reasons. Eighty-three per cent (83.0%) were wholly satisfied with the consultation. The consultation scored an average of 8.3, SD 1.6, measured on a visual analogue scale of 0-10. When asked the most important aspects of the consultation 54.6% cited treatment, 25.4% reassurance and 20.0% diagnosis. Ninety-six per cent felt that their treatment had been adequately explained; however, 78.3% made errors when reporting their regimen. A significant improvement in patients' level of recall was found after they had received further information from the hospital pharmacist. Firstly, this study shows patients' perceptions and priorities when visiting eye casualty. Secondly, it demonstrates that patients are generally satisfied with their eye casualty attendance. Thirdly, many patients depart with poor understanding of their eye treatment regime which is likely to affect compliance. Communication between doctors and patients was enhanced by involvement of the hospital pharmacist. This strategy is applicable not only to an ophthalmic casualty unit but also to a wider range of settings and could provide a service standard for future audit.


Why do more divergent sequences produce smaller nonsynonymous/synonymous rate ratios in pairwise sequence comparisons?
Several studies have reported a negative correlation between estimates of the nonsynonymous to synonymous rate ratio (ω = dN/dS) and the sequence distance d in pairwise comparisons of the same gene from different species. That is, more divergent sequences produce smaller estimates of ω. Explanations for this negative correlation have included segregating nonsynonymous polymorphisms in closely related species and nonlinear dynamics of the ratio of two random variables. Here we study the statistical properties of the maximum-likelihood estimates of ω and d in pairwise alignments and explore the possibility that the negative correlation can be entirely explained by those properties. We show that the ω estimate is positively biased for small d and that the bias decreases with the increase of d. We also show that the estimates of ω and d are negatively correlated when ω < 1 and positively correlated when ω > 1. However, the bias in estimates of ω and the correlation between estimates of ω and d are not enough to explain the much stronger correlation observed in real data sets. We then explore the behavior of the estimates when the model is misspecified and suggest that the observed correlation may be due to protein-level selection that causes very different amino acids to be favored in different domains of the protein. Widely used models fail to account for such among-site heterogeneity and cause underestimation of the nonsynonymous rate and ω, with the bias being much stronger for distant sequences. We point out that tests of positive selection based on the ω ratio are invariant to the parameterization of the model and thus unaffected by bias in the ω estimates or the correlation between estimates of ω and d.


Discovery of G-quadruplex-forming sequences in SARS-CoV-2.
The outbreak caused by the novel coronavirus SARS-CoV-2 has been declared a global health emergency. G-quadruplex structures in genomes have long been considered essential for regulating a number of biological processes in a plethora of organisms. We have analyzed and identified 25 four contiguous GG runs (G2NxG2NyG2NzG2) in the SARS-CoV-2 RNA genome, suggesting putative G-quadruplex-forming sequences (PQSs). Detailed analysis of SARS-CoV-2 PQSs revealed their locations in the open reading frames of ORF1 ab, spike (S), ORF3a, membrane (M) and nucleocapsid (N) genes. Identical PQSs were also found in the other members of the Coronaviridae family. The top-ranked PQSs at positions 13385 and 24268 were confirmed to form RNA G-quadruplex structures in vitro by multiple spectroscopic assays. Furthermore, their direct interactions with viral helicase (nsp13) were determined by microscale thermophoresis. Molecular docking model suggests that nsp13 distorts the G-quadruplex structure by allowing the guanine bases to be flipped away from the guanine quartet planes. Targeting viral helicase and G-quadruplex structure represents an attractive approach for potentially inhibiting the SARS-CoV-2 virus.


[Granular cell laryngeal tumor].
Granular cell tumor is a rare benign tumor which when affects the larynx, it characteristically affects the vocal cords, causing dysphonia as the main clinical manifestation. The term granular cell myoblastoma proposed and extensively used seems to be inadequate since all the observations suggest its origin in Schwann cells, with no relation to muscle cells. Although the existence of malignant granular cell tumors has been reported, the vast majority show a benign behaviour, reaching its cure after complete surgical removal of the lesion.


[Ophthalmological symptoms of measles and their treatment].
Measles, caused by the Morbilli virus, is a highly (about 95 %) contagious disease affecting primarily children, but without proper immunisation, adults can also be infected. The leading symptoms of the disease are high fever that presents after an incubation period of 9-10 days and the red rash that begins several days after the fever starts. Beyond specific generalized symptoms, measles may have ocular symptoms. The most commonly occurring conjunctivitis, the so-called "red eye symptom", is not characteristic only for measles infection, however, by taking the generalized symptoms it can suggest the diagnosis at the beginning of the disease. Conjunctivitis of varying severity is noticed in the half of the cases without using ophthalmological instrumentation. Using ophthalmological instrumentation, the mild forms of conjunctivitis can be diagnosed, by meticulous ophthalmological examination, further eye diseases can be discovered. The viral conjunctivitis can progress to keratitis and bacterial superinfection can occur. If the infection presents in childhood it can affect the posterior segment. The fight against measles is very effective in Hungary since the vaccination has been introduced, and the lack of vaccination is also the primary cause of the risk to the disease. In the diagnosis, symptomatic treatment of the disease and the curbing of possible mass infections, the practicing physician (general practitioner) has a key role. The correct care of the infected patient in Hungary is provided by a methodological letter, professional information and legal guides. Orv Hetil. 2017; 158(39): 1523-1527.


Suppression of Escherichia coli formate hydrogenlyase activity by trimethylamine N-oxide is due to drainage of the inducer formate.
The effect of the addition of trimethylamine N-oxide (TMAO) in the growth medium on Escherichia coli anaerobic fermentative and respiratory pathways was examined. Formate dehydrogenase H (FDH-H) activity was totally repressed by the addition of 40 mM TMAO, whereas the overall hydrogenase (HYD) activity was reduced by 25%. Accordingly, expression of lacZ operon fusions with the fdhF and hycB structural genes specifying FDH-H and HYD3 was reduced sevenfold and eightfold, respectively, leading to suppression of an active formate hydrogenlyase system. In contrast, global respiratory formate-dependent phenazine methosulphate reductase (FDH-PMS) activity, which consists of both the major anaerobic FDH-N enzyme and the aerobic FDH-Z isoenzyme, was increased approximately twofold. This was corroborated by a 2.5-fold stimulation of the sole fdoG-uidA transcriptional fusion which reflects the synthesis of the respiratory aerobic FDH-Z enzyme. In fdhD, fdhE or torA mutants lacking either FDH-PMS activity or TMAO reductase (TOR) activity, the formate hydrogenlyase pathway was no longer inhibited by TMAO. In addition, introduction of 30 mM formate in the growth medium was found to relieve the repressive effect of TMAO in the wild-type strain. When TMAO was added as terminal electron acceptor a significant enhancement of anaerobic growth was observed with the wild-type strain and the fdoG mutant. It was associated with the concomitant suppression of the formate hydrogenlyase enzymes. This was in contrast to the fdnG and torA mutants whose growth pattern and fermentative enzymes remained unaffected. Taken together, these results strongly suggest that formate-dependent reduction of TMAO via FDH-N and TOR reduces the amount of formate available for induction of the formate hydrogenlyase pathway.


A phosphorylation site in the Na+ channel required for modulation by protein kinase C.
Voltage-gated sodium channels are responsible for generation of action potentials in excitable cells. Activation of protein kinase C slows inactivation of sodium channels and reduces peak sodium currents. Phosphorylation of a single residue, serine 1506, that is located in the conserved intracellular loop between domains III and IV and is involved in inactivation of the sodium channel, is required for both modulatory effects. Mutant sodium channels lacking this phosphorylation site have normal functional properties in unstimulated cells but do not respond to activation of protein kinase C. Phosphorylation of this conserved site in sodium channel alpha subunits may regulate electrical activity in a wide range of excitable cells.


Development and survivorship of Lycorma delicatula (Hemiptera: Fulgoridae) on cultivated and native Vitis spp. (Vitales: Vitaceae) of the Eastern United States.
As Lycorma delicatula (White) continues to spread across the United States, more winegrapes are potentially susceptible to damage from this pest. Lycorma delicatula, spotted lanternfly, is primarily associated with Ailanthus altissima (Mill.) Swingle, a tree from its native range that is now globally distributed. While L. delicatula is a known pest of cultivated Vitis spp. in South Korea, its relationship with the specific grape species grown in the United States is unclear. This study assessed L. delicatula survivorship and development on 5 Vitis species, including 2 winegrape V. vinifera L. varieties, 'Pinot Noir' and 'Chardonnay', Concord grape, Vitis labrusca L., River grape, Vitis riparia Michx., and muscadine grape, Vitis rotundifolia Michx. var. 'Carlos'. A diet of A. altissima served as a positive control. Lycorma delicatula provided with a diet of V. riparia or V. vinifera 'Pinot Noir' yielded the highest survivorship and fastest rates of development among grape diets and were statistically equivalent to those provided with A. altissima. Vitis rotundifolia did not support L. delicatula growth past the third-instar life stage, indicating this species is a poor host for the early development of this pest. Our results indicate that both V. riparia and V. vinifera are favorable hosts for L. delicatula and may provide the means for this insect to invade and establish in new regions.


Potassium transport in cortical collecting tubules from mineralocorticoid-treated rat.
Unidirectional fluxes of 86Rb+ were used to examine the stimulation of K+ secretion produced by arginine vasopressin (ADH) in isolated perfused cortical collecting tubules from rats treated with desoxycorticosterone. ADH at 100 microU/ml in the bathing solution increased the bath-to-lumen flux (Jb----l; pmol X min-1 X mm-1) from 16.9 +/- 2.3 to 43.2 +/- 4.6 (n = 16). The lumen-to-bath flux (Jl----b) fell from 3.2 +/- 0.7 to 1.3 +/- 0.4 with ADH due to hyperpolarization of the transepithelial voltage from -12.6 +/- 1.3 to -39.3 +/- 2.0 mV, but the calculated Rb+ permeability was unaltered at 0.20-0.26 micron/s. Although 2 mM lumen Ba2+ inhibited Jb----l by 55 +/- 6%, the flux ratio (Jb----l/Jl----b) of 28 +/- 8 was larger than predicted for passive exchange. In the absence of ADH 2 mM Ba2+ reduced Jb----l to the level predicted for passive movement, but addition of ADH with Ba2+ still present increased Jb----l by an amount identical to that observed without Ba2+, although the absolute Jb----l was less. Simultaneous addition of 2 mM luminal and 4 mM bath Ba2+ also inhibited Jl----b for 22Na+ but not to passive levels. These results indicate either that the concentrations of Ba2+ used were insufficient to block K+ conductances completely or that K+/Rb+ secretion can occur through a Ba2+-insensitive pathway.


Multipotent mesenchymal stromal cells enhance insulin secretion from human islets via N-cadherin interaction and prolong function of transplanted encapsulated islets in mice.
Multipotent mesenchymal stromal cells (MSC) enhance viability and function of islets of Langerhans. We aimed to examine the interactions between human MSC and human islets of Langerhans that influence the function of islets. Human MSC and human islets (or pseudoislets, obtained after digestion and reaggregation of islet cells) were cocultured with or without cellular contact and glucose-stimulated insulin secretion assays were performed to assess cell function. The expression of several adhesion molecules, notably ICAM-1 and N-cadherin on islets and MSC, was investigated by qPCR. The role of N-cadherin was analyzed by adding an anti-N-cadherin antibody in islets cultured with or without MSC for 24 h followed by insulin measurements in static incubation assays. Islets and MSC were coencapsulated in new hydrogel microspheres composed of calcium alginate and covalently crosslinked polyethylene glycol. Encapsulated cells were transplanted intraperitoneally in streptozotocin-induced diabetic mice and glycemia was monitored. Islet function was evaluated by the intraperitoneal glucose tolerance test. In vitro, free islets and pseudoislets cocultured in contact with MSC showed a significantly increased insulin secretion when compared to islets or pseudoislets cultured alone or cocultured without cell-to-cell contact with MSC (p < 0.05). The expression of ICAM-1 and N-cadherin was present on islets and MSC. Blocking N-cadherin prevented the enhanced insulin secretion by islets cultured in contact with MSC whereas it did not affect insulin secretion by islets cultured alone. Upon transplantation in diabetic mice, islets microencapsulated together with MSC showed significantly prolonged normoglycemia when compared with islets alone (median 69 and 39 days, respectively, p < 0.01). The intraperitoneal glucose tolerance test revealed an improved glycemic response in mice treated with islets microencapsulated together with MSC compared to mice transplanted with islets alone (p < 0.001). MSC improve survival and function of islets of Langerhans by cell-to-cell contact mediated by the adhesion molecule N-cadherin.


Role of extracellular hemoglobin in thrombosis and vascular occlusion in patients with sickle cell anemia.
Sickle cell anemia (SCA) is a common hemolytic disorder caused by a gene mutation in the β-globin subunit of hemoglobin (Hb) and affects millions of people. The intravascular hemolysis releases excessive amount of extracellular hemoglobin (ECHb) into plasma that causes many cellular dysfunctions in patients with SCA. ECHb scavenges NO which promotes crisis events such as vasoconstriction, thrombosis and hypercoagulation. ECHb and its degradation product, heme, are known to cause oxidative damage to the vessel wall and stimulate the expression of adhesive protein ligands on vascular endothelium. Our study shows that ECHb binds potently to VWF-largest multimeric glycoprotein in circulation-through the A2-domain, and significantly inhibits its cleavage by the metalloprotease ADAMTS13. Furthermore, a subpopulation of VWF multimers bound to ECHb exists in significant amount, accounting for about 14% of total plasma VWF, in SCD patients. The Hb-bound VWF multimers are resistant to ADAMTS13, and are hyperactive in aggregating platelets. Thus, the data suggest that Hb-bound VWF multimers are ultralarge and hyperactive because they are resistant to the protease. The Hb-bound VWF multimers are elevated parallely with the level of ECHb in patients' plasma, and is associated with the pathogenesis of thrombosis and vascular occlusion in SCA.


Autologous stem cell transplantation for enteropathy-associated T-cell lymphoma: a retrospective study by the EBMT.
Enteropathy-associated T-cell lymphoma (EATL) is a rare subtype of peripheral T-cell lymphomas with a poor prognosis. Autologous stem cell transplantation (ASCT) was retrospectively evaluated as a consolidation or salvage strategy for EATL. The analysis included 44 patients who received ASCT for EATL between 2000 and 2010. Thirty-one patients (70%) were in first complete or partial remission at the time of the ASCT. With a median follow-up of 46 months, relapse incidence, progression-free survival, and overall survival were 39%, 54%, and 59% at 4 years, respectively, with only one relapse occurring beyond 18 months posttransplant. There was a trend for better survival in patients transplanted in first complete or partial remission at 4 years (66% vs 36%; P = .062). ASCT is feasible in selected patients with EATL and can yield durable disease control in a significant proportion of the patients.


Making workplaces safer: The influence of organisational climate and individual differences on safety behaviour.
Current work health and safety practices focus predominately on fostering a safety climate to promote safety behaviours and reduce workplace accidents. Despite the importance of safety climates in accident prevention, recent research has demonstrated that individual factors can also predict work safety behaviour. This study considered the importance of organisational climate together with individual characteristics including differences in personality, impulsiveness, and perceptions of safety within the workplace on safety behaviour. 203 participants consisting of 67 males and 136 females aged 18 to 71 years, completed an online questionnaire. Results revealed that safety behaviour was directly related to safety climate, and conscientiousness. In contrast, neuroticism, and impulsiveness were not significantly related to safety behaviour. The present study findings support previous findings in the literature regarding the importance of safety climate as well as the personality trait of conscientiousness in applying safety behaviours. However, the present study findings did not support previous research in relation to the personality trait of high neuroticism resulting in decreased safety behaviour, nor did not confirm an inverse relationship between high impulsivity and low safety behaviour as theoretical models would suggest. This new finding may warrant further research into the precursors for safety behaviour.


[Mast cells and heparin--key links in adaptive and pathological processes].
Participation of mast cells and heparin in adaptive and pathological processes is discussed with special reference to heparin pool in mast cell granules. Hypotheses explaining conversion of normal mast cells to phenotypically pathologic ones are analysed. Prospects for enhancement of adaptive abilities of the organism and approaches to the treatment of diseased organs are discussed in terms of protection and restoration of heparin cells in the mast cells.


Dysregulation of gene expression in ABCC6 knockdown HepG2 cells.
ABCC6 protein is an ATP-dependent transporter that is mainly found in the basolateral plasma membrane of hepatocytes. ABCC6 deficiency is the primary cause of several forms of ectopic mineralization syndrome. Mutations in the human ABCC6 gene cause pseudoxanthoma elasticum (PXE), an autosomal recessive disease characterized by ectopic calcification of the elastic fibers in dermal, ocular and vascular tissues. Mutations in the mouse ABCC6 gene were also associated with dystrophic cardiac calcification. Reduced levels of ABCC6 protein were found in a β-thalassemic mouse model. Moreover, some cases of generalized arterial calcification in infancy are due to ABCC6 mutations. In order to study the role of ABCC6 in the pathogenesis of ectopic mineralization, the expressions of genes involved in this process were evaluated in HepG2 cells upon stable knockdown of ABCC6 by small hairpin RNA (shRNA) technology. ABCC6 knockdown in HepG2 cells causes a significant upregulation of the genes promoting mineralization, such as TNAP, and a parallel downregulation of genes with anti-mineralization activity, such as NT5E, Fetuin A and Osteopontin. Although the absence of ABCC6 has been already associated with ectopic mineralization syndromes, this study is the first to show a direct relationship between reduced ABCC6 levels and the expression of pro-mineralization genes in hepatocytes.


Value-driven attentional capture enhances distractor representations in early visual cortex.
When a behaviorally relevant stimulus has been previously associated with reward, behavioral responses are faster and more accurate compared to equally relevant but less valuable stimuli. Conversely, task-irrelevant stimuli that were previously associated with a high reward can capture attention and distract processing away from relevant stimuli (e.g., seeing a chocolate bar in the pantry when you are looking for a nice, healthy apple). Although increasing the value of task-relevant stimuli systematically up-regulates neural responses in early visual cortex to facilitate information processing, it is not clear whether the value of task-irrelevant distractors influences behavior via competition in early visual cortex or via competition at later stages of decision-making and response selection. Here, we measured functional magnetic resonance imaging (fMRI) in human visual cortex while subjects performed a value-based learning task, and we applied a multivariate inverted encoding model (IEM) to assess the fidelity of distractor representations in early visual cortex. We found that the fidelity of neural representations related to task-irrelevant distractors increased when the distractors were previously associated with a high reward. This finding suggests that value-driven attentional capture begins with sensory modulations of distractor representations in early areas of visual cortex.


High-frequency transfer of cloned herpes simplex virus type 1 sequences to mammalian cells by protoplast fusion.
The protoplast fusion technique of Schaffner (W. Schaffner, Proc. Natl. Acad. Sci. U.S.A. 77:2163-2167, 1980) has been adapted to introduce cloned herpes simplex virus genes into cultured mammalian cells. The technique involves digesting bacterial cell walls with lysozyme to produce protoplasts and then fusing the protoplasts to mammalian cells by treatment with polyethylene glycol. For monitoring transfer, protoplasts were labeled with the fluorescent dye fluorescein isothiocyanate before fusion. After fusion, greater than 50% of the mammalian cells were fluorescent, demonstrating that bacterial material was transferred with high frequency. Transfer of plasmid pBR325 occurred at frequencies of 1 to 2%, as measured by in situ hybridization. Fusion transfer of a chimeric plasmid consisting of the herpes simplex virus type 1 (strain KOS) EcoRI fragment F in pBR325 resulted in expression of some viral genomic sequences in about 5% of the mammalian cells, as detected by indirect immunofluorescence. One Ltk- cell in 300 to 500 was transformed to the TK+ phenotype after fusion with protoplasts carrying the chimeric plasmid pX1, which consists of pBR322 and the BamHI fragment coding for the herpes simplex virus type 1 thymidine kinase gene.


[Congenital malformations among offspring of diabetic women].
The risk of congenital malformations is two to three times higher among women with diabetes mellitus before pregnancy. Gestational diabetes is emerging as a risk factor for malformations. To study the rate and type of congenital malformations among offspring of women with pre-gestational or gestational diabetes. A search in the databases of the Collaborative Latin American Study for Congenital Malformations and of the Neonatology Unity of The University of Chile Clinical Hospital. All births from January 1998 through June 2004 were investigated. In the study period, 13,965 births occurred, of which 0.7% were stillbirths and 8.1% had malformations. A total of 295 women (2.1%) had diabetes (gestational diabetes in 1.8% and pre gestational diabetes in 0.3%). The prevalence of congenital malformations was 14% and 18.3% among offspring of mothers with pre-gestational and gestational diabetes, respectively (p=NS). The prevalence of congenital malformations among offspring of non diabetic women was 7.9% (p <0.001 with respect to diabetic women). Major malformations were found in 83% and 70% of malformed offspring of women with pre-gestational and gestational diabetes, respectively. Pregnant women with pre-existing or gestational diabetes have a higher risk of giving birth a child with malformations.


Clinical implications of gene dosage and gene expression patterns in diploid breast carcinoma.
Deregulation of key cellular pathways is fundamental for the survival and expansion of neoplastic cells. In cancer, regulation of gene transcription can be mediated in a variety of ways. The purpose of this study was to assess the impact of gene dosage on gene expression patterns and the effect of other mechanisms on transcriptional levels, and to associate these genomic changes with clinicopathologic parameters. We screened 97 invasive diploid breast tumors for DNA copy number alterations and changes in transcriptional levels using array comparative genomic hybridization and expression microarrays, respectively. The integrative analysis identified an increase in the overall number of genetic alterations during tumor progression and 15 specific genomic regions with aberrant DNA copy numbers in at least 25% of the patient population, i.e., 1q22, 1q22-q23.1, 1q25.3, 1q32.1, 1q32.1-q32.2, 8q21.2-q21.3, 8q22.3, 8q24.3, and 16p11.2 were recurrently gained, whereas 11q25, 16q21, 16q23.3, and 17p12 were frequently lost (P < 0.01). An examination of the expression patterns of genes mapping within the detected genetic aberrations identified 47 unique genes and 1 Unigene cluster significantly correlated between the DNA and relative mRNA levels. In addition, more malignant tumors with normal gene dosage levels displayed a recurrent overexpression of UBE2C, S100A8, and CBX2, and downregulation of LOC389033, STC2, DNALI1, SCUBE2, NME5, SUSD3, SERPINA11, AZGP1, and PIP. Taken together, our findings suggest that the dysregulated genes identified here are critical for breast cancer initiation and progression, and could be used as novel therapeutic targets for drug development to complement classical clinicopathologic features.


Secretion of type IV collagenolytic protease and metastatic phenotype: induction by transfection with c-Ha-ras but not c-Ha-ras plus Ad2-E1a.
Activated ras oncogene transfection into suitable recipient cells has been shown to induce the metastatic phenotype (Thorgeirsson, et al., Mol. Cell. Biol., 5: 259-262, 1985). We have used this model system to study the correlation of basement membrane collagenolysis with metastatic propensity. The c-Ha-ras oncogene alone, or combined with v-myc, transfected into early passage rat embryo fibroblasts, induce these cells to secrete high levels of type IV collagenolytic metalloproteinase and to concomitantly exhibit a high incidence of spontaneous metastases in nude mice. Cotransfection of c-Ha-ras plus the adenovirus type 2 E1a gene yields cells which are highly tumorigenic but nonmetastatic and fail to produce type IV collagenase. This effect is due to a suppression of collagenase elaboration, not increased production of a collagenase inhibitor, and not decreased production of a collagenase activator. The characteristics of the collagenase are identical to tumor type IV collagenase described previously. The nonmetastatic cells which failed to produce type IV collagenase retain the ability to secrete high levels of plasminogen activator. Transfection with the protooncogenic forms of Ha-ras or mos, or spontaneous transformation of NIH 3T3 cells or chemical transformation of BALB 3T3 cells yields cells which fail to produce collagenase, are tumorigenic, but totally nonmetastatic. These data support a biochemical linkage of type IV collagenase expression with the metastatic phenotype in this rodent system.


Kaposi Sarcoma in the Era of Rapamycin Remains a Therapeutic Challenge in Organ Transplant Recipients.
Solid-organ transplant recipients are at higher risk of developing Kaposi sarcoma, which is a multicentric vascular neoplasm of lymphatic endothelium-derived cells. Reducing doses of immunosuppressive drugs and switching from calcineurin inhibitors to the mammalian target of rapamycin inhibitor rapamycin have been suggested as an effective first-line treatment modality in most patients. Herein, we report a 64-year-old renal transplant recipient who developed multiple cutaneous and visceral Kaposi sarcoma lesions 2 months after transplant. The patient showed no improvement, with progression of the disease until month 15 of the suggested therapy of rapamycin.


Learning peptide recognition rules for a low-specificity protein.
Many proteins interact with short linear regions of target proteins. For some proteins, however, it is difficult to identify a well-defined sequence motif that defines its target peptides. To overcome this difficulty, we used supervised machine learning to train a model that treats each peptide as a collection of easily-calculated biochemical features rather than as an amino acid sequence. As a test case, we dissected the peptide-recognition rules for human S100A5 (hA5), a low-specificity calcium binding protein. We trained a Random Forest model against a recently released, high-throughput phage display dataset collected for hA5. The model identifies hydrophobicity and shape complementarity, rather than polar contacts, as the primary determinants of peptide binding specificity in hA5. We tested this hypothesis by solving a crystal structure of hA5 and through computational docking studies of diverse peptides onto hA5. These structural studies revealed that peptides exhibit multiple binding modes at the hA5 peptide interface-all of which have few polar contacts with hA5. Finally, we used our trained model to predict new, plausible binding targets in the human proteome. This revealed a fragment of the protein α-1-syntrophin that binds to hA5. Our work helps better understand the biochemistry and biology of hA5, as well as demonstrating how high-throughput experiments coupled with machine learning of biochemical features can reveal the determinants of binding specificity in low-specificity proteins.


[Controlled arterial hypotension using nitroglycerin in brain surgery].
Artificial hypotension induced by 0.01% nitroglycerin solution was applied in 60 operations on the brain. It contributed to the creation of optimum conditions for radical removal of tumors and exclusion of vascular aneurysms, reduction of blood loss, and a decrease in the number of blood transfusion. The efficacy, adequate control, safety, and attainability of the method allowed it to be recommended for use in neurosurgical practice when necessary.


Identification of chlorophylls and carotenoids in major teas by high-performance liquid chromatography with photodiode array detection.
The separation and identification of pigments, chlorophylls, and carotenoids of seven teas and fresh leaf of tea (Camellia sinensis) by high-performance liquid chromatography (HPLC) are described. HPLC was carried out using a Symmetry C(8) column with a photodiode array detector. Pigments were eluted with a binary gradient of aqueous pyridine solution at a flow rate of 1.0 mL/min at 25 degrees C. HPLC analyses achieved the separation of more than 100 pigment peaks, and 79 pigment species, 41 chlorophylls, and 38 carotenoids were detected. The presence of degraded chlorophylls was a common feature, and the number and the variety of pigments differed with tea species. Generally, the numbers of chlorophyll species tended to increase with processing steps, while carotenoid species were decreased, especially by heating. Particularly in green teas, a change of carotenoid structure, conversion of violaxanthin to auroxanthin, occurred. In hot water extracts of teas, both chlorophylls and carotenoids were also detected, but the concentration of chlorophylls was less than 2% as compared with acetone extracts. The pigment compositions were compared between tea species, and they are discussed in terms of the differences in their manufacturing processes.


Possible role for cytotoxic oxygen metabolites in the pathogenesis of cardiac ischemic injury.
Parabiotically perfused, isolated cat hearts were subjected to global ischemia for 2 hours at 27 degrees C and were then reperfused for 2 hours. One group of hearts was simply perfused with hypothermic blood 5 minutes before ischemia and again before normothermic reperfusion. A second group received a crystalloid cardioplegic solution. A third group received cardioplegic solution supplemented with superoxide dismutase and catalase; at the start of reperfusion the anesthetized cats supplying blood to perfused hearts in this group received an i.v. infusion of these enzymes. Based on comparisons of postreperfusion ventricular pressure development, maximal ventricular dP/dt, rate-pressure products and coronary blood flow, we concluded that cardioplegic solution and enzyme treatment were superior to enzyme-free cardioplegia solution under otherwise similar experimental conditions. The results suggest that a component of "ischemic" cardiac damage may involve cytotoxicity from oxygen metabolites such as superoxide anion, hydrogen peroxide, or both, and that this component of damage can be reduced by enzyme supplements such as superoxide dismutase or catalase.


Management of low bone mineral density in premenopausal women.
To review evidence for management of low bone density in premenopausal women and to establish practical guidelines for management of low bone density in this population by family physicians. A search of MEDLINE for relevant articles published between January 1990 and May 2004 was conducted. Articles retrieved were graded by level of evidence. Recommendations for diagnosis and therapy were based on evidence from randomized controlled trials and expert consensus. Low bone density in premenopausal women is not associated with the same increased risk of fracture seen in older women. In the absence of fragility fractures and loss of height, it may be a reflection of low peak bone mass and may represent the normal variation in bone mineral density (BMD). Women may have low bone density secondary to an underlying skeletal or systemic disorder. Common causes of low bone density in premenopausal women include ovulatory disturbances and low body weight. Osteoporosis is diagnosed in the premenopausal female population in the presence of fragility fractures and is not based solely on the results of BMD testing. Secondary causes of bone loss should be excluded, and any underlying condition contributing to low bone density should be corrected. Antiresorptive therapy has been evaluated only in those premenopausal women who are on glucocorticoid therapy and in those with primary hyperparathyroidism. Only in these conditions has antiresorptive therapy been shown to improve BMD.


Studying the Functional Potential of Ground Ivy (Glechoma hederacea L.) Extract Using an In Vitro Methodology.
Glechoma hederacea L., known as ground ivy, has a long history of use in folk medicine. The main bioactive compounds in ground ivy are polyphenolic compounds known for their potent antioxidant and antimicrobial activities and thus have high potential as functional ingredients against bacterial infections and the occurrence of chronic diseases associated with oxidative stress in the human body. The aim of the present study was to determine the biological activity of ground ivy extract on selected human cell lines, including hepatic (HepG2), tongue (CAL 27), gastric (AGS) and colon (Caco-2) cancer cell lines by evaluating cytotoxicity, formation of reactive oxygen species and genotoxicity. The antioxidant capacity of the extract was additionally evaluated using cellular model macromolecules of protein and DNA, bovine serum album and plasmid phiX174 RF1 DNA. The effect of ground ivy extract on representatives of human microflora, including L. plantarum, E. coli and S. aureus, was also studied. The cytotoxicity of the extract depended on the type of cells treated, and the pro-oxidant effect generally decreased with increasing exposure time. The most pronounced genoprotective effect against hydroxyl radical damage was monitored in model plasmid DNA and occurred at the highest tested concentration (0.25 mg mL-1), with 95.89% preservation of the supercoiled form of the plasmid. This concentration also had the most significant antioxidant activity on the model protein-14.01% more than the positive control prepared using Trolox. The ground ivy extract showed high antimicrobial potential against the pathogenic bacteria E. coli and S. aureus.


Topological Structural Brain Connectivity Alterations in Aspartylglucosaminuria: A Case-Control Study.
We investigated global and local properties of the structural brain connectivity networks in aspartylglucosaminuria, an autosomal recessive and progressive neurodegenerative lysosomal storage disease. Brain connectivity in aspartylglucosaminuria has not been investigated before, but previous structural MR imaging studies have shown brain atrophy, delayed myelination, and decreased thalamic and increased periventricular WM T2 signal intensity. We acquired diffusion MR imaging and T1-weighted data from 12 patients with aspartylglucosaminuria (mean age, 23 [SD, 8] years; 5 men), and 30 healthy controls (mean age, 25 [SD, 10] years; 13 men). We performed whole-brain constrained spherical deconvolution tractography, which enables the reconstruction of neural tracts through regions with complex fiber configurations, and used graph-theoretical analysis to investigate the structural brain connectivity networks. The integration of the networks was decreased, as demonstrated by a decreased normalized global efficiency and an increased normalized characteristic path length. In addition, the average strength of the networks was decreased. In the local analyses, we found decreased strength in 11 nodes, including, for example, the right thalamus, right putamen, and, bilaterally, several occipital and temporal regions. We found global and local structural connectivity alterations in aspartylglucosaminuria. Biomarkers related to the treatment efficacy are needed, and brain network properties may provide the means for long term follow-up.


Reconstitution of Monomethylamine:Coenzyme M methyl transfer with a corrinoid protein and two methyltransferases purified from Methanosarcina barkeri.
Methanogenesis from methylamines requires the intermediate methylation of 2-mercaptoethanesulfonate (CoM). In vitro reconstitution of CoM methylation with monomethylamine was achieved with three purified proteins: a monomethylamine corrinoid protein (MMCP), the "A" isozyme of methylcobamide:CoM methyltransferase (MT2-A), and a newly isolated protein termed monomethylamine methyltransferase (MMAMT).MMAMT is a 170-kDa protein with 52-kDa subunits. The MMAMT polypeptide was rate-limiting for methyl transfer until at a 2-fold molar excess over MMCP. MMAMT is a monomethylamine:MMCP methyltransferase, since methylation of MMCP required MMAMT but not MT2-A. MMCP and MMAMT formed a complex detectable by size exclusion high pressure liquid chromatography. Methyl group transfer from methyl-MMCP to CoM was mediated by MT2-A, since methyl iodide:CoM methyl transfer by MMCP and MT2-A did not require MMAMT. MT2-M, an isozyme of MT2-A, was inactive in MMCP-dependent methyl transfer. Immunodepletion of MMCP from the extract inhibited CoM methylation with monomethylamine but not dimethylamine. Purified MMCP reconstituted activity in immunodepleted extracts. These results show that MMCP is the major corrinoid protein for methanogenesis from monomethylamine detectable in extracts and that it interacts with two methyltransferases. MMAMT functions as a MMA:MMCP methyltransferase, while MT2-A functions as a methyl-MMCP:CoM methyltransferase.


A double-blind comparison of intranasal budesonide with placebo for nasal polyposis.
Intranasal budesonide, 400 micrograms two times a day, was evaluated in 36 patients referred for treatment of nasal polyposis. The age range was 20 to 68 years. Polypectomy was done 5.6 (mean) times previously. After a 5-week, treatment-free, baseline period, patients were treated in a double-blind fashion with either budesonide or placebo during 4 weeks. After this treatment period, placebo-treated patients started receiving budesonide in an open trial for an additional 4 weeks. The patients rated their nasal symptoms daily. Nasal examinations and nasal inspiratory flow rate (IFR) measurements were done at clinic visits. After 3 and 4 weeks of treatment, the response to budesonide was significantly greater than response to placebo. The greater reduction in nasal blockage caused by polyps, observed on physical examination, p = 0.005, was mirrored by an increase in nasal IFR (p = 0.0001). Patient rating of the severity and frequency of nasal blockage were reduced more by budesonide than by placebo (p less than or equal to 0.0005). Switching placebo-treated patients to budesonide treatment resulted in a reduction of nasal blockage (p less than 0.001) and an increase in nasal IFR (p less than 0.001). The results demonstrate that topical nasal budesonide, 400 micrograms two times a day, is an effective treatment of nasal polyps.


[Study of the role of parents' occupation in children's mortality].
This study have been carried out to: a) investigate the prevalence of such occupational factors in parents which could be fatal to their children, and b) evaluate quantitatively the risk of mortality in infants caused by some leading diseases developed as a result of harmful occupational factors to which their parents had been exposed. During this work the data bank on infant mortality in Tallinn based on deaths registration in city archive for period 1968-1992 has been created. This bank includes the data on parents' occupation. The prevalence of parents' occupational risk factors has been analyzed in infants who die due extreme immaturity and other prematurity cases, congenital anomalies neoplasias and control group persons. To evaluate the death risk in children the case-control epidemiological study with the individual group selection has been performed. This pilot analytical epidemiological study allows to suppose involving of possibly harmful occupational factors which are playing a certain role influencing mortality in whole contingent investigated, but not concerning individual nosologic forms. For the final judgement concerning the role of occupational risk factors the additional special fundamental study including the extended number of observations and the data, on interrupting nonoccupational risk factors prevalence should be carried out.


Haploidentical transplantation.
When allogeneic transplant is indicated and a MSD is not available, most centers first seek an unrelated donor. However, one must avoid undue delays when a donor is not available and family members should be typed to identify the preferred donor amongst the family. Since a family donor can provide access to transplant for almost every patient, a greater effort should be encouraged to improve and expand haploidentical transplantation. The advantages, outlined in Table 1, suggest compelling reason to perfect the use of haploidentical related donors.


Melatonin Uptake by Cells: An Answer to Its Relationship with Glucose?
Melatonin, N-acetyl-5-methoxytryptamine, is an indole mainly synthesized from tryptophan in the pineal gland and secreted exclusively during the night in all the animals reported to date. While the pineal gland is the major source responsible for this night rise, it is not at all the exclusive production site and many other tissues and organs produce melatonin as well. Likewise, melatonin is not restricted to vertebrates, as its presence has been reported in almost all the phyla from protozoa to mammals. Melatonin displays a large set of functions including adaptation to light: dark cycles, free radical scavenging ability, antioxidant enzyme modulation, immunomodulatory actions or differentiation⁻proliferation regulatory effects, among others. However, in addition to those important functions, this evolutionary 'ancient' molecule still hides further tools with important cellular implications. The major goal of the present review is to discuss the data and experiments that have addressed the relationship between the indole and glucose. Classically, the pineal gland and a pinealectomy were associated with glucose homeostasis even before melatonin was chemically isolated. Numerous reports have provided the molecular components underlying the regulatory actions of melatonin on insulin secretion in pancreatic beta-cells, mainly involving membrane receptors MTNR1A/B, which would be partially responsible for the circadian rhythmicity of insulin in the organism. More recently, a new line of evidence has shown that glucose transporters GLUT/SLC2A are linked to melatonin uptake and its cellular internalization. Beside its binding to membrane receptors, melatonin transportation into the cytoplasm, required for its free radical scavenging abilities, still generates a great deal of debate. Thus, GLUT transporters might constitute at least one of the keys to explain the relationship between glucose and melatonin. These and other potential mechanisms responsible for such interaction are also discussed here.


Controlling nonsequential double ionization via two-color few-cycle pulses.
Using the classical three-dimensional ensembles, we have demonstrated the controlling of dynamics in nonsequential double ionization (NSDI) of helium by two-color few-cycle pulses. By changing the relative phase of the two pulses, recollisions leading to NSDI can be restricted in a time interval of several hundreds attosecond nearly before the field extremum and as a result, the correlated electron momentum distribution exhibits a so-far unobserved narrow arc-like structure. This structure reveals a novel energy correlation between the two electrons from NSDI by two-color few-cycle pulses.


Entamoeba histolytica: early progressive pathology in the cecum of the gerbil (Meriones unguiculatus).
The early formation of amebic cecal lesions in the gerbil (Meriones unguiculatus) was studied sequentially from 1 to 10 days post-inoculation. The earliest response of the cecal mucosa towards trophozoites of Entamoeba histolytica was local release of interglandular and goblet cell mucin. Trophozoites adhered to the surface epithelium or were trapped in the luminal mucus in the absence of epithelial cell damage. Focal mucosal erosion occurred later, and was preceded by crypt hyperplasia, edema and hemorrhaging of the lamina propria. The interglandular epithelial cells were cytolysed or were sloughed off into the lumen. Amebic mucosal erosion was accompanied by trophozoite entry in the crypts, but rarely into the lamina propria. The inflammatory cellular infiltrate was diffuse, and neutrophils were found in the lumen in contact with trophozoites. The findings of this study suggest that trophozoites of E. histolytica do not actively penetrate or invade the interglandular epithelium, but rather, that amebae are permitted to invade as a result of the destruction of the epithelium. Mucus depletion and the mucosal inflammation, probably in response to the adsorption of amebic toxins, preceded epithelial erosions.


A Preliminary Study on qEEG in Burn Patients With Chronic Pruritus.
To explore and determine the reorganizational changes in the cortical neural circuits associated with pruritis, this study was undertaken to compare the electroencephalography (EEG) changes in burn patients having primary symptoms of chronic itching (pruritis) and their paired healthy subjects. Eight subjects were recruited for this exploratory pilot study: 4 patients with pruritus after burn injury matched by gender and age with 4 healthy subjects. EEG recordings were analyzed for absolute alpha, low beta, high beta, and theta power for both groups. The mean age of the burn patients was 41.75 years; while the mean age for the matched healthy subjects was 41.5 years. All subjects were male. A decreased alpha activity was observed in the occipital channels (0.82 vs. 1.4; p=0.01) and a decreased low beta activity in the frontal area (0.22 vs. 0.4; p=0.049) in eyes closed conditions. An overall decreased theta trend was observed in both the eyes open and eyes closed conditions in burn patients, compared to healthy individuals. This preliminary study presents initial evidence that chronic pruritus in burn subjects may be associated with brain reorganizational changes at the cortical level characterized by an EEG pattern.


Determination of nickel in water by chemiluminescence.
The acetone chemiluminescence determination of nickel in water was investigated. Optimization data for the determination of Ni2+ and interference data for over 20 species are provided. The limit of detection for Ni2+ by this method is 2.5 ng ml-1, and the linear dynamic range is from 10 ng ml-1 to 1 microgram ml-1. The application of the method to the determination of Ni2+ in river and waste waters is discussed.


Lung cancer incidence decreases with elevation: evidence for oxygen as an inhaled carcinogen.
The level of atmospheric oxygen, a driver of free radical damage and tumorigenesis, decreases sharply with rising elevation. To understand whether ambient oxygen plays a role in human carcinogenesis, we characterized age-adjusted cancer incidence (compiled by the National Cancer Institute from 2005 to 2009) across counties of the elevation-varying Western United States and compared trends displayed by respiratory cancer (lung) and non-respiratory cancers (breast, colorectal, and prostate). To adjust for important demographic and cancer-risk factors, 8-12 covariates were considered for each cancer. We produced regression models that captured known risks. Models demonstrated that elevation is strongly, negatively associated with lung cancer incidence (p < 10(-16)), but not with the incidence of non-respiratory cancers. For every 1,000 m rise in elevation, lung cancer incidence decreased by 7.23 99% CI [5.18-9.29] cases per 100,000 individuals, equivalent to 12.7% of the mean incidence, 56.8. As a predictor of lung cancer incidence, elevation was second only to smoking prevalence in terms of significance and effect size. Furthermore, no evidence of ecological fallacy or of confounding arising from evaluated factors was detected: the lung cancer association was robust to varying regression models, county stratification, and population subgrouping; additionally seven environmental correlates of elevation, such as exposure to sunlight and fine particulate matter, could not capture the association. Overall, our findings suggest the presence of an inhaled carcinogen inherently and inversely tied to elevation, offering epidemiological support for oxygen-driven tumorigenesis. Finally, highlighting the need to consider elevation in studies of lung cancer, we demonstrated that previously reported inverse lung cancer associations with radon and UVB became insignificant after accounting for elevation.


A web-based survey on various symptoms of computer vision syndrome and the genetic understanding based on a multi-trait genome-wide association study.
A variety of eye-related symptoms due to the overuse of digital devices is collectively referred to as computer vision syndrome (CVS). In this study, a web-based survey about mind and body functions, including eye strain, was conducted on 1998 Japanese volunteers. To investigate the biological mechanisms behind CVS, a multi-trait genome-wide association study (GWAS), a multivariate analysis on individual-level multivariate data, was performed based on the structural equation modeling methodology assuming a causal pathway for a genetic variant to influence each symptom via a single common latent variable. Twelve loci containing lead variants with a suggestive level of significance were identified. Two loci showed relatively strong signals and were associated with TRABD2B relative to the Wnt signaling pathway and SDK1 having neuronal adhesion and immune functions, respectively. By utilizing publicly available eQTL data, colocalization between GWAS and eQTL signals for four loci was detected, and a locus on 2p25.3 showed a strong colocalization (PPH4 > 0.9) on retinal MYT1L, known to play an important role in neuronal differentiation. This study suggested that the use of multivariate questionnaire data and multi-trait GWAS can lead to biologically reasonable findings and enhance our genetic understanding of complex relationships among symptoms related to CVS.


Stimulus statistics shape oscillations in nonlinear recurrent neural networks.
Rhythmic activity plays a central role in neural computations and brain functions ranging from homeostasis to attention, as well as in neurological and neuropsychiatric disorders. Despite this pervasiveness, little is known about the mechanisms whereby the frequency and power of oscillatory activity are modulated, and how they reflect the inputs received by neurons. Numerous studies have reported input-dependent fluctuations in peak frequency and power (as well as couplings across these features). However, it remains unresolved what mediates these spectral shifts among neural populations. Extending previous findings regarding stochastic nonlinear systems and experimental observations, we provide analytical insights regarding oscillatory responses of neural populations to stimulation from either endogenous or exogenous origins. Using a deceptively simple yet sparse and randomly connected network of neurons, we show how spiking inputs can reliably modulate the peak frequency and power expressed by synchronous neural populations without any changes in circuitry. Our results reveal that a generic, non-nonlinear and input-induced mechanism can robustly mediate these spectral fluctuations, and thus provide a framework in which inputs to the neurons bidirectionally regulate both the frequency and power expressed by synchronous populations. Theoretical and computational analysis of the ensuing spectral fluctuations was found to reflect the underlying dynamics of the input stimuli driving the neurons. Our results provide insights regarding a generic mechanism supporting spectral transitions observed across cortical networks and spanning multiple frequency bands.


Arrays of pressure sensors based on organic field effect: a new perspective for non invasive monitoring.
In this paper we propose totally flexible organic field effect transistors (OFETs) assembled on plastic films as sensors for mechanical variables. In the first part, mechanical sensors for pressure and bending detection are presented. A sharp and reversible sensitivity of the output current of the device to an elastic deformation induced by means of a mechanical stimulus on the device channel has been observed and suggested the idea of employing arrays of such sensors for detecting the deformation applied onto a planar surface. In the second part, the possibility of using similar devices for bio- and chemo-detection is described. By exploiting the properties of the basic structure, the device can be combined with any kind of substrate to detect for instance the pressure applied by people walking or standing on a functionalized carpet. This emerging technology seems to be promising for applications in the field of remote and non invasive monitoring of elderly and disabled people.


Pathophysiology of early trauma-induced coagulopathy: emerging evidence for hemodilution and coagulation factor depletion.
Trauma patients present with a coagulopathy, termed early trauma-induced coagulopathy (ETIC), that is associated with increased mortality. This study investigated hemostatic changes responsible for ETIC. Case-control study of trauma patients with and without ETIC, defined as prolonged prothrombin time (PT), was performed from prospective cohort of consecutive trauma patients who presented to Level I trauma center. Univariate and multivariate analyses were performed. The case-control study group (n = 91) was 80% male, with mean age of 37 years, 17% penetrating trauma and 7% mortality rate. Patients with ETIC demonstrated decreased common and extrinsic pathway factor activities (factors V and VII) and decreased inhibition of the coagulation cascade (antithrombin and protein C activities) when compared with the matched control patients without ETIC. Both cohorts had evidence of increased thrombin and fibrin generation (prothrombin fragment 1.2 levels, thrombin-antithrombin complexes, and soluble fibrin monomer), increased fibrinolysis (d-dimer levels), and increased inhibition of fibrinolysis (plasminogen activator inhibitor-1 activity) above normal reference values. Patients with versus without ETIC had increased mortality and received increased amount of blood products. ETIC following injury is associated with decreased factor activities without significant differences in thrombin and fibrin generation, suggesting that despite these perturbations in the coagulation cascade, patients displayed a balanced hemostatic response to injury. The lower factor activities are likely secondary to increased hemodilution and coagulation factor depletion. Thus, decreasing the amount of crystalloid infused in the early phases following trauma and administration of coagulation factors may prevent the development.


Relationship between hyperoxemia and ventilator associated pneumonia.
Previous studies suggest a relationship between hyperoxemia and ventilator-associated pneumonia (VAP). Hyperoxemia is responsible for denitrogenation phenomena, and inhibition of surfactant production, promoting atelectasis in mechanically ventilated patients. Further, hyperoxemia impairs the efficacy of alveolar macrophages to migrate, phagocyte and kill bacteria. Oxygen can also cause pulmonary-specific toxic effect called hyperoxic acute lung injury leading to longer duration of mechanical ventilation. All these hyperoxic effects are well-known risk factors for VAP. A recent retrospective large single center study identified hyperoxemia as an independent risk factor for VAP. However, two recent randomized controlled trials evaluated the impact of conservative oxygen strategy versus a liberal strategy, but did not confirm the role of hyperoxemia in lower respiratory tract infection occurrence. In this review, we discuss animal and human studies suggesting a relationship between these two common conditions in mechanically ventilated patients and potential interventions that should be evaluated. Further large prospective studies in carefully selected groups of patients are required to confirm the potential role of hyperoxemia in VAP pathogenesis and to evaluate the impact of a conservative oxygen strategy vs. a conventional strategy on the incidence of VAP.


Glass Microsphere-Supported Giant Vesicles for the Observation of Self-Reproduction of Lipid Boundaries.
Growth and division experiments on phospholipid boundaries were carried out using glass microsphere-supported phospholipid (DOPC) giant vesicles (GVs) fed with a fatty acid solution (oleic acid) at two distinct feeding rates. Both fast and slow feeding methods produced daughter GVs. Under slow feeding conditions the membrane growth process (evagination, buds, filaments) was observed in detail by fluorescence microscopy. The density difference between supported mother vesicles and newly formed daughter vesicles allowed their easy separation. Mass spectrometric analysis of the resulting mother and daughter GVs showed that the composition of both vesicle types was a mixture of original supported phospholipids and added fatty acids reflecting the total composition of amphiphiles after the feeding process. Thus, self-reproduction of phospholipid vesicles can take place under preservation of the lipid composition but different aggregate size.


Stent retriever use for retrieval of displaced microcoils: a consecutive case series.
Coil displacement during endovascular coiling procedures may require coil retrieval in the context of flow limitation or thromboembolic risk. No standard recommended method of coil retrieval exists. We present a consecutive series of 14 patients with displaced coil during aneurysm coiling in whom the complication was effectively managed with the use of a stent retriever system. Two illustrative cases from the 14 are described, and technical notes are detailed regarding use of the technique. The use of stent retrievers presents a simple, safe, and effective choice for removal of prolapsed coils during aneurysm coiling.


Transcript CD81-215 may be a long noncoding RNA of stromal origin with tumor-promoting role in colon cancer.
The role of tetraspanin CD81 in malignant transformation is best studied in colorectal cancer, and it appears that other transcripts beside the fully coding mRNA may also be dysregulated in malignant cells. Recent data from a comprehensive pan-cancer transcriptome analysis demonstrated differential activity of two alternative CD81 gene promoters in malignant versus nonmalignant gut mucosa. The promoter active in gut mucosa gives rise to transcripts CD81-203 and CD81-213, while the promoter active in colon and rectal cancer gives rise to transcripts CD81-205 and CD81-215. Our study aimed to explore the biomarker potential of the transcripts from the alternative CD81 gene promoters in colon cancer, as well as to investigate their structure and potential function using in silico tools. The analysis of the transcripts' expression in several colon cell lines cultivated in 2D and 3D and a set of colon cancer and healthy gut mucosa samples by qPCR and RNA sequencing suggested their low expression and stromal origin. Expression patterns in tumor and nontumor tissue along with in silico data suppose that the transcript CD81-215 may be a noncoding RNA of stromal origin with possible involvement in signaling related to malignant transformation.


Modification of the adhesive properties of silicone-based coatings by block copolymers.
The improvement of the (bio)adhesive properties of elastomeric polydimethylsiloxane (PDMS) coatings is reported. This is achieved by a surface modification consisting of the incorporation of block copolymers containing a PDMS block and a poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) block in a PDMS matrix, followed by matrix cross-linking and immersion of the obtained materials in water. Contact angle measurements (CA), X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM) showed the presence of the PDMAEMA block at the surface, drastic morphology changes, and improved adhesion properties after immersion in water. Finally, underwater bioadhesion tests show that mussels adhere only to block copolymer-filled coatings and after immersion in water, i.e., when the PDMAEMA blocks have been brought to the coating surface. These observations highlight the significant role of hydrophilic groups in the surface modification of silicone coatings.


Consideration of Future Consequences Influences Involvement in Romantic Partners' Health.
This study examined the influence of people's short-term (e.g., convenience) and long-term (e.g., maintain weight) motives toward their romantic partner's health on their provision of health-related social control and autonomy support for their partner's health behaviors, as well as their indifference toward their romantic partner's health. Data from a sample of cohabiting couples (N = 212) were analyzed using multilevel path models. Results showed having more long-term motives for a partner's health lead to increased provisions of autonomy support and social control, and less indifference toward the partner's health, especially when the partner had lower long-term health motives themselves. In contrast, having more short-term motives for a partner's health was associated with providing less social control and autonomy support and showing more indifference toward the partner's health. Thus, people motivated to improve their partner's long-term health engage in more behaviors meant to improve their romantic partner's health choices, particularly when the partner has low motivation to regulate their own health behaviors.


Adapting an ambient monitoring program to the challenge of managing emerging pollutants in the San Francisco Estuary.
While over seven million organic and inorganic compounds that have been indexed by the American Chemical Society's Chemical Abstracts Service in their CAS Registry are commercially available, most pollution monitoring programs focus only on those chemical stressors for which regulatory benchmarks exist, and have been traditionally considered responsible for the most significant human and environmental health risks. Until the late 1990s, the San Francisco Estuary Regional Monitoring Program was no exception in that regard. After a thorough external review, the monitoring program responded to the need for developing a pro-active surveillance approach for emerging pollutants in recognition of the fact that the potential for the growing list of widely used chemical compounds to alter the integrity of water is high. We describe (1) the scientific and analytical bases underlying a new surveillance monitoring approach; (2) summarize approaches used and results obtained from a forensic retrospective; (3) present the growing data set on emerging pollutants from surveillance monitoring and related efforts in the San Francisco Bay Area to characterize newly targeted compounds in wastewater streams, sediment, storm water runoff, and biota; and (4) suggest next steps in monitoring program development and applied research that could move beyond traditional approaches of pollutant characterization. Based on the forensic analysis of archived chromatograms and chemical and toxicological properties of candidate compounds, we quantified a variety of synthetic organic compounds which had previously not been targeted for analysis. Flame retardant compounds, pesticides and insecticide synergists, insect repellents, pharmaceuticals, personal care product ingredients, plasticizers, non-ionic surfactants, and other manufacturing ingredients were detected in water, sediment, and/or biological tissue samples. Several of these compounds, especially polybrominated diphenyl ether flame retardants, exhibited concentrations of environmental concern. We also describe environmental management challenges associated with emerging pollutants and how pro-active surveillance monitoring might assist in implementing a more holistic approach to pollution prevention and control before emerging pollutants become a burden on future generations.


Medical students' knowledge, attitude, and practice of complementary and alternative medicine: a pre-and post-exposure survey in Majmaah University, Saudi Arabia.
Evidently, complementary and alternative medicine (CAM) is a recognized medical practice that efficiently uses multiple treatment therapies and techniques in the prevention and management of a variety of human disorders. Many medical schools have integrated CAM curriculum in medical education system worldwide. Research in knowledge, attitude, and practice (KAP) of diverse health professionals exposed to CAM courses is important from many perspectives including improvement in KAP and teaching skills of faculty, together with capacity building and curriculum development. This pre- and post-design cross-sectional study aimed to assess CAM-KAP of two intakes of medical students in Majmaah University, Saudi Arabia. The second-year medical students of the first (year 2012-2013) and second (year 2013-2014) intake (n=26 and 39, respectively) were selected for this study. A reliable, 16-item self-administered questionnaire was distributed among all the students for answering before and after the 48-hour CAM course. The data were analyzed using appropriate statistical test of significance. Medical students' knowledge and attitude toward CAM significantly improved across some subitems of CAM questionnaire with a positive trend in the rest of its items including their views on CAM practices. CAM course tends to have a positive impact on KAP of medical students. The preliminary results of this study call for further research with a larger sample in academic settings across the nation.


An Unusual Acute Coronary Syndrome Due to a Septic Embolism: A Case Presentation and Review of Revascularization Strategies.
Treatment of acute coronary syndromes secondary to septic coronary valvular embolism in endocarditis patients remains unclear. Several revascularization strategies have been described, including thromboaspiration, stent implantation, balloon angioplasty, and surgical intervention. We herein present an illustration of an atypical case of an acute coronary syndrome related to a coronary bifurcation occlusion due to a septic embolism in a patient presenting with infective endocarditis. We also summarized previous similar cases and their management.


New cellular markers at diagnosis are associated with isolated central nervous system relapse in paediatric B-cell precursor acute lymphoblastic leukaemia.
In childhood acute lymphoblastic leukaemia (ALL), central nervous system (CNS) involvement is rare at diagnosis (1-4%), but more frequent at relapse (~30%). Because of the significant late sequelae of CNS treatment, early identification of patients at risk of CNS relapse is crucial. Using microarray-analysis, we discovered multiple differentially expressed genes between B-cell precursor (BCP) ALL cells in bone marrow (BM) and BCP-ALL cells in cerebrospinal fluid (CSF) at the time of isolated CNS relapse. After confirmation by real-time quantitative polymerase chain reaction, selected genes (including SCD and SPP1) were validated at the protein level by flowcytometric analysis of BCP-ALL cells in CSF. Further flowcytometric validation showed that a subpopulation of BCP-ALL cells (>1%) with a 'CNS protein profile' (SCD positivity and increased SPP1 expression) was present in the BM at diagnosis in patients who later developed an isolated CNS relapse, whereas this subpopulation was <1% or absent in all other patients. These data indicate that the presence of a (small) subpopulation of BCP-ALL cells with a 'CNS protein profile' at diagnosis (particularly SCD-positivity) is associated with isolated CNS relapse. Such information can be used to design new diagnostic and treatment strategies that aim at prevention of CNS relapse with reduced toxicity.


Serotonin depletion in vivo inhibits the branching of olfactory projection neurons in the lobster deutocerebrum.
Serotonin depletion during embryogenesis has been shown previously to retard the growth of the olfactory and accessory lobes of the lobster deutocerebrum (Benton et al., 1997). The present study was undertaken to determine whether morphological changes in the interneurons innervating these lobes contribute to this growth retardation. We examined the effects of in vivo serotonin depletion using 5,7-dihydroxytryptamine (5,7-DHT) on the morphology of the olfactory projection neurons, one of two major classes of interneurons that innervate both lobes. Intracellular dye fills of olfactory projection neurons in normal embryos showed that each neuron extensively innervates either the olfactory or accessory lobe before projecting to neuropil regions in the protocerebrum. In embryos injected with 5,7-DHT, however, the deutocerebral arbors of 13.5% of the olfactory projection neurons examined were either markedly reduced compared with normal neurons or absent. Affected neurons also exhibited a number of additional aberrant morphological features suggesting that these neurons represent cells that were affected during their initial morphogenesis. Olfactory projection neurons with aberrant morphologies were also encountered, although less frequently (7.5% of the neurons examined), in control (sham-injected) embryos indicating that the sham injections can affect the development of the brain. This observation provides insights into the nature of effects seen in control embryos in previous experiments (Benton et al., 1997). The results of the present study indicate that in vivo serotonin depletion inhibits the branching of olfactory projection neurons and suggest, therefore, that one of the functions of serotonin during normal development is to promote the ingrowth of these neurons into the deutocerebral neuropils.


Human leukocyte antigen and cytokine receptor gene polymorphisms associated with heterogeneous immune responses to mumps viral vaccine.
Mumps outbreaks continue to occur throughout the world, including in highly vaccinated populations. Vaccination against mumps has been successful; however, humoral and cellular immune responses to mumps vaccines vary significantly from person to person. We set out to assess whether HLA and cytokine gene polymorphisms are associated with variations in the immune response to mumps viral vaccine. To identify genetic factors that might contribute to variations in mumps vaccine-induced immune responses, we performed HLA genotyping in a group of 346 healthy schoolchildren (12-18 years of age) who previously received 2 doses of live mumps vaccine. Single-nucleotide polymorphisms (minor allele frequency of >5%) in cytokine and cytokine receptor genes were genotyped for a subset of 118 children. Median values for mumps-specific antibody titers and lymphoproliferative stimulation indices were 729 IU/mL and 4.8, respectively. Girls demonstrated significantly higher mumps antibody titers than boys, indicating gender-linked genetic differences in humoral immune response. Significant associations were found between the HLA-DQB1*0303 alleles and lower mumps-specific antibody titers. An interesting finding was the association of several HLA class II alleles with mumps-specific lymphoproliferation. Alleles of the DRB1 (*0101, *0301, *0801, *1001, *1201, and *1302), DQA1 (*0101, *0105, *0401, and *0501), and DQB1 (*0201, *0402, and *0501) loci were associated with significant variations in lymphoproliferative immune responses to mumps vaccine. Additional associations were observed with single-nucleotide polymorphisms in the interleukin-10RA, interleukin-12RB1, and interleukin-12RB2 cytokine receptor genes. Minor alleles for 4 single-nucleotide polymorphisms within interleukin-10RA and interleukin-12RB genes were associated with variations in humoral and cellular immune responses to mumps vaccination. These data suggest the important role of HLA and immunoregulatory cytokine receptor gene polymorphisms in explaining variations in mumps vaccine-induced immune responses.


Pyk2 regulates cell-edge protrusion dynamics by interacting with Crk.
Focal adhesion kinase (FAK) is well established as a regulator of cell migration, but whether and how the closely related proline-rich tyrosine kinase 2 (Pyk2) regulates fibroblast motility is still under debate. Using mouse embryonic fibroblasts (MEFs) from Pyk2-/- mice, we show here, for the first time, that lack of Pyk2 significantly impairs both random and directed fibroblast motility. Pyk2-/- MEFs show reduced cell-edge protrusion dynamics, which is dependent on both the kinase and protein-protein binding activities of Pyk2. Using bioinformatics analysis of in vitro high- throughput screens followed by text mining, we identified CrkI/II as novel substrates and interactors of Pyk2. Knockdown of CrkI/II shows altered dynamics of cell-edge protrusions, which is similar to the phenotype observed in Pyk2-/- MEFs. Moreover, epistasis experiments suggest that Pyk2 regulates the dynamics of cell-edge protrusions via direct and indirect interactions with Crk that enable both activation and down-regulation of Crk-mediated cytoskeletal signaling. This complex mechanism may enable fine-tuning of cell-edge protrusion dynamics and consequent cell migration on the one hand together with tight regulation of cell motility, a process that should be strictly limited to specific time and context in normal cells, on the other hand.


FcR gamma presence in TCR complex of double-negative T cells is critical for their regulatory function.
TCRalphabeta+CD4-CD8- double-negative (DN) T regulatory (Treg) cells have recently been shown to suppress Ag-specific immune responses mediated by CD8+ and CD4+ T cells in humans and mice. Our previous study using cDNA microarray analysis of global gene expression showed that FcRgamma was the most highly overexpressed gene in functional DN Treg cell clones compared with nonfunctional mutant clones. In this study, we demonstrate that FcRgamma-deficient DN T cells display markedly reduced suppressive activity in vitro. In addition, unlike FcRgamma-sufficient DN T cells, FcRgamma-deficient DN T cells were unable to prolong donor-specific allograft survival when adoptively transferred to recipient mice. Protein analyses indicate that in addition to FcRgamma, DN Treg cell clones also express higher levels of TCRbeta, while mutant clones expressed higher levels of Zap70 and Lck. Within DN Treg cells, we found that FcRgamma associates with the TCR complex and that both FcRgamma and Syk are phosphorylated in response to TCR cross-linking. Inhibition of Syk signaling and FcRgamma expression were both found to reduce the suppressive function of DN Treg cells in vitro. These results indicate that FcRgamma deficiency significantly impairs the ability of DN Treg cells to down-regulate allogeneic immune responses both in vitro and in vivo, and that FcRgamma plays a role in mediating TCR signaling in DN Treg cells.