Newborn Screening for Congenital Hypothyroidism-Clinical Evaluation and Comparison of Two Different Test Kits for the Determination of TSH in Dried Blood Samples on Two Different Platforms.
Newborn screening (NBS) for congenital hypothyroidism (CH) started in the 1970s, with the introduction of radioimmuno assays (RIA) for the measurement of thyroxine (T4), and thyroid stimulating hormone (TSH). With the development of sensitive enzyme immune assays (EIA, FIA, FEIA), RIAs were replaced in the newborn screening laboratories. With the increasing number of analytes and centralization of NBS, there is a growing demand of total automation. In the course of method validation, two fully automated platforms for the determination of TSH in dried blood samples (DBS) were compared. The GSP from PerkinElmer (PE), and the NS2400 from Labsystems (LDx), together with the recommended test kits from both manufacturers. Both systems showed good performance, with recoveries, of 103.0% (LDx) and 98.5% (PE), and CVs for intra and interassay variations at various concentrations, between 4.3 and 15.7. Both assays had a good correlation (r2 = 0.8814). With LDx/NS2400 platform, TSH values were in the mean 2.09 mU/L higher; however, the difference of both results from the mean was within ±2 SD, up to 30 mU/L, and only for values above 50 mU/L did the difference become bigger. However, this has no influence on the clinical interpretation. No false negative results were observed with either of the two platforms. TSH results obtained with the LDx/NS2400 were slightly higher than those obtained with the PE/GSP; however, the recall rate was lower: 0.059% compared to 0.063%. This can be explained by the much narrower distribution of TSH values. In conclusion, both platforms are equally suitable for medium and large NBS laboratories. However, due to the more open structure the LDx/NS2400 platform has a lot of advantages compared to the totally closed PE/GSP platform.


Encoding activity in the medial temporal lobe examined with anatomically constrained fMRI analysis.
Functional neuroimaging studies have produced a sizable number of observations of increased activity in the human medial temporal lobe (MTL) during encoding of novel memories. The studies have suggested possible functional specialization within the anatomical components of the MTL (hippocampus and the entorhinal, perirhinal, and parahippocampal cortical areas). Neuroimaging studies have just begun to link anatomical regions to specific functions. To address functional specialization hypothesis, a method is described for using high-resolution structural information from magnetic resonance imaging MRI to constrain the analysis of functional magnetic resonance imaging (fMRI) data, for independent assessment of functional activity change in each component of the MTL. With this method, increased activity was detected throughout the MTL in a group of participants (n = 5) who encoded novel pictures. A separate group (n = 5) who encoded words exhibited lower-levels of evoked activity. Laterality effects were found reflecting increased right hemisphere activity during picture encoding (parahippocampal cortex) and increased left hemisphere activity during word encoding (posterior hippocampus and parahippocampal cortex). Neither condition provided evidence for greater activity in the posterior hippocampus than in the anterior hippocampus during encoding, although the greatest increases in activity were observed in the parahippocampal cortex. The anatomically driven methodology is shown to provide detailed comparison of levels of activity change across specific brain areas and to provide increased sensitivity to functional change in each region of the MTL.


Biglycan gene expression in UVR-exposed rabbit corneas.
It is known that stromal proteoglycans play an important role in the hydration and transparency of the mammalian cornea. Proteoglycans have been described as a pathological deposit in climatic proteoglycan stromal keratopathy, which is associated with chronic ultraviolet radiation (UVR) exposure. The expression of dermatan sulfate proteoglycan biglycan in the cornea was thus studied after exposure of rabbit eyes to UVR. New Zealand albino rabbit corneas were exposed to UVR at 310 nm at the dose producing biomicroscopically significant keratitis (0.47 J/cm2). Animals were killed 3, 7 and 28 days after exposure (five rabbits in each group). Five rabbits were used as controls and did not receive any UVR treatment. Expression of biglycan mRNA in the corneas was investigated by competitive reverse transcription-polymerase chain reaction (RT-PCR). There was no expression of biglycan mRNA in the control group. In the UVR-exposed groups, biglycan mRNA had still not been expressed 3 days after exposure. The expression of biglycan mRNA was observed in all UVR-treated corneas 7 days after exposure (p < 0.05). By 28 days after UVR exposure the expression of biglycan mRNA had decreased (not statistically significant). There is no detectable biglycan gene expression in the normal rabbit cornea. Ultraviolet radiation exposure leads to a distinct expression of biglycan mRNA in the rabbit cornea that decreases 4 weeks after exposure, indicating the involvement of biglycan in the corneal repair process. Biglycan appears to be a novel marker of corneal wound healing.


[Work satisfaction and absenteeism of nursing staff--comparative study of 1021 nurse trainees and nurses].
To analyse the high level of absenteeism among nursing trainees compared with nursing staff. Unlike previous studies, the present study focussed on work satisfaction and motivation. Specifically, combining satisfaction with absenteeism was a novel approach. For assessing work satisfaction, a standardised form with 73 items in four areas was drafted and checked in a pre-test (n = 150). 861 nurses and 159 trainees were interviewed. The absenteeism data given by the nursing staff were compared with the 'missing' records of the personnel department. In all areas it was found that, in particular, problems of organisation, personnel management and working atmosphere in the hospital were a burden on the employees. In detail, however, there were considerable differences between nurses and trainees in respect of appraisal. Work organisation: Although trainees rated work organisation aspects lower than nurses, direct relationship to work satisfaction was less pronounced. For the trainees, improvements are imperative in respect of active self-responsibility. Leadership/co-operation: Trainees rated supervisor behaviour and working atmosphere lower than their colleagues. There was a direct relation to satisfaction and absenteeism. Workload/stress: Although their responsibility was less, a larger proportion of the trainees felt stressed. This was directly related to work satisfaction and absenteeism. Fluctuation and turnover: 44% of the trainees would be prepared to work up to the age of retirement, but only 25% of the qualified staff. Nevertheless, three-quarters of the trainees and two-thirds of the nurses would choose the same profession again. Hence, unfavourable local (internal) circumstances led to the discontent and not the profession as such. The extremely high absenteeism of nursing trainees calls for action on the part of school and hospital management. There is a need for better information and care before and during professional training, because workload will be higher at the end of the training. The study indicates possibilities of increasing work satisfaction and decreasing absenteeism of trainees.


ToxDB: pathway-level interpretation of drug-treatment data.
Extensive drug treatment gene expression data have been generated in order to identify biomarkers that are predictive for toxicity or to classify compounds. However, such patterns are often highly variable across compounds and lack robustness. We and others have previously shown that supervised expression patterns based on pathway concepts rather than unsupervised patterns are more robust and can be used to assess toxicity for entire classes of drugs more reliably. We have developed a database, ToxDB, for the analysis of the functional consequences of drug treatment at the pathway level. We have collected 2694 pathway concepts and computed numerical response scores of these pathways for 437 drugs and chemicals and 7464 different experimental conditions. ToxDB provides functionalities for exploring these pathway responses by offering tools for visualization and differential analysis allowing for comparisons of different treatment parameters and for linking this data with toxicity annotation and chemical information.Database URL:http://toxdb.molgen.mpg.de.


Attachment of Paramecium to polystyrene surfaces: a model system for the analysis of sexual cell recognition and nuclear activation.
In the ciliate Paramecium caudatum, only mating-reactive cells can attach to the polystyrene surface of Petri dishes. The attachment of mating-reactive cells occurs exclusively at the tips of ventrally located cilia (mating-reactive cilia) and appears to involve a hydrophobic interaction. An increase in the hydrophilic nature of the polystyrene surface following treatment with sulphuric acid results in the loss of attachment of reactive cells. The presence or absence of Ca ions and varying the ionic strength of the medium have no effect upon attachment. Although attachment reactivity is closely correlated with mating reactivity, the attachment sites appear distinct from the mating recognition sites, as suggested by the following experimental results: (I) cells that lose mating reactivity following treatment with trypsin are still able to attach to the polystyrene surface; (2) the attachment of cells appears temperature-dependent over a range where mating reactivity remains uneffected; (3) mating reactivity is quickly lost as a result of attachment to the surface of the dish. While the mechanism of attachment is not the same as sexual recognition, the attachment to the polystyrene surface initiates the early stages of the conjugation process. Cells that attach show micronuclear migration while unattached cells do not. The maximum rate of micronuclear migration occurs approximately 30 min after attachment. Meiosis does not occur following migration in the attached cells as in cells that are sexually mating. The relationship between the mating substance and the attachment substance is discussed, while the utilization of the attachment phenomenon for other experimental procedures is proposed.


Circulating parathyroid hormone and serotonin in multiparous cows with differing postparturient serum calcium concentrations.
Many multiparous dairy cows experience subclinical hypocalcemia (SCH) in the immediate postpartum period as they adapt to the demands of lactation. Furthermore, differing dynamics of SCH in the days following parturition are associated with varied health and production outcomes, with cows experiencing transient SCH producing more milk and facing fewer negative health events than cows with delayed or persistent SCH. Our objectives were to describe differences in mediators of calcium (Ca) homeostasis between cows experiencing differing Ca dynamics postpartum. A prospective cohort of 89 multiparous Holstein cows from 2 herds in NY were classified into 1 of 4 SCH groups based on mean serum total Ca (tCa) at 1 and 4 DIM: normocalcemic (NC; [tCa] > 1.89 mmol/L at 1 DIM and > 2.25 mmol/L at 4 DIM, n = 30); transient SCH (tSCH; [tCa] ≤ 1.89 mmol/L at 1 DIM and > 2.25 mmol/L at 4 DIM, n = 12); delayed SCH (dSCH; [tCa] > 1.89 mmol/L at 1 DIM and ≤ 2.25 mmol/L at 4 DIM, n = 23); and persistent SCH (pSCH; [tCa] ≤ 1.89 mmol at 1 DIM and ≤ 2.25 mmol/L at 4 DIM, n = 24). Blood samples were collected at -5, -1, 1 through 5, 7, and 10 DIM and analyzed for tCa, parathyroid hormone (PTH), and serotonin. Repeated measures ANOVA models were used to analyze differences between SCH groups and changes over time for tCa, PTH, and serotonin. During the prepartum period, tCa was greater in the NC, tSCH, and dSCH cows as compared with the pSCH cows and there was marginal evidence for a difference in PTH between SCH groups. Postpartum tCa varied over time between SCH groups. Mean postpartum tCa for respective SCH groups were: NC = 2.32 mmol/L, 95% confidence interval (2.28, 2.35) mmol/L, tSCH = 2.20 (2.14, 2.25) mmol/L, dSCH = 2.17 (2.13, 2.21) mmol/L, and pSCH = 2.03 (1.99, 2.07) mmol/L. Mean concentrations of PTH in the postpartum period were: NC = 70.1 (66.2, 74.4) pmol/L, tSCH = 72.1 (66.1, 79.2), dSCH = 75.8 (70.8, 81.5) pmol/L, and pSCH = 77.7 (72.4, 83.9) pmol/L. Serotonin was similar between SCH groups pre- and postpartum and followed a cyclical pattern from 1 to 10 DIM. Our results agreed with our hypothesis that differences in postpartum PTH might exist between cows experiencing different dynamics of SCH in the early lactation period; however, further studies are needed to confirm this difference. If true, this would suggest that Ca homeostasis may be disrupted in cows with dSCH and pSCH. Gaining a better understanding of these modulatory differences may aid in the prevention, management, and treatment of SCH.


Oesophageal carcinoma associated with corrosive injury--prevention and treatment by oesophageal resection.
Six patients (5 from Kuala Lumpur and 1 from Singapore), who had a history of corrosive injury in late adolescence and early adult life, developed oesophageal squamous cell carcinoma. The association between corrosion and carcinoma was probably causal in 5 women who developed oesophageal cancer 25-50 years after corrosive injury and coincidental in a 35-year-old man with a recent history of corrosive ingestion. The 5 patients from Kuala Lumpur were among a series of 49 patients treated over a 10-year period for dysphagia from corrosive injury. Excluding the patient with coincidental association, the 4 patients with corrosive-induced carcinoma represent an 8 per cent incidence of malignancy in the series of 49 patients. Reference is made to the safety of oesophageal resection in 28 patients with associated carcinoma. Oesophageal resection during reconstructive surgery for severe corrosive stricture would have a preventive role in corrosive induced carcinoma. However, these cases suggest that patients with relatively mild corrosive injury not requiring reconstructive surgery and also at risk of oesophageal carcinoma. An awareness of this risk should lead to earlier diagnosis of carcinoma and an increase in the number of curative resections.


Early Embryogenesis and Anterior-Posterior Axis Formation in the White-Tip Nematode Aphelenchoides besseyi (Nematoda: Aphelenchoididae).
We followed the early embryogenesis of Aphelenchoides besseyi from fertilization to the 4-cell stage under Nomarski optics and examined the chromosome number and structure by DAPI staining. After an oocyte is fertilized by a sperm, the eggshell forms and the male and female pronuclei are reconstructed. The male pronucleus moves toward the female pronucleus, which is located at the center of the egg. They meet, rotate 90°, and fuse. The embryo then divides unequally into a larger anterior AB cell and a smaller posterior P(1) cell. The site of sperm entry into the oocyte seems to become the future anterior pole of the embryo, and thus the formation of an anterior-posterior axis formation is the same as that for Bursaphelenchus xylophilus, but opposite to that for Caenorhabditis elegans. From immunostaining, the fertilizing sperm appears to bring the centrosome into the oocyte. The chromosome structure during the pronuclear meeting as observed by DAPI staining suggests that a haploid sperm (N = 3) fertilizes a haploid oocyte (N = 3) to form a diploid embryo (2N = 6) and that all chromosomes appear to be of a similar size. Unlike C. elegans does, the P(1) cell first divides anterior-posteriorly followed by the AB anterior-posteriorly. These divisions produced the 4-cell stage embryo with 4 cells arranged in a linear fashion, again in contrast to that for C. elegans or B. xylophilus configured in a rhomboid shape.


[Preparation and characterization of a hemostatic porous platelet-rich plasma chitosan/silk fibroin wound dressing].
In order to further improve the hemostatic performance of wound dressings, human platelet-rich plasma (PRP) containing various growth factors was introduced into chitosan solution. The silk fibroin solutions with different volume ratios (1:1, 1:3, 3:1 and 1:0) were added to improve the porosity and hemostasis of materials. The hPRP-chitosan/silk wound dressings with different ratios was prepared by freeze-drying and pure chitosan dressing was considered as the control group to study the effects of PRP and silk fibroin in different proportions on the hemostasis properties and the growth factors burst release. The hemostasis of chitosan dressing was improved by introducing PRP, but the porous structure and water absorption were not significantly improved. If silk fibroin solution was added in the ratio of 1:1, the more uniform porous structure and better hemostatic performance could be obtained. The porosity and water absorption could reach 86.83%±3.84% and 1 474%±114% respectively. In addition, the dressings with ratio of 1:1 had positive effects on reducing the burst release of growth factors on initial stage. Therefore, PRP-chitosan/silk fibroin composite dressing can become a kind of wound dressing that can achieve rapid hemostasis and promote wound healing.


Isolated Torsion of Accessory Fallopian Tube in a Young Adolescent.
Accessory fallopian tube is a rare anatomical variation, which might rarely cause gynecological complications such as infertility, ectopic pregnancy, cystic swelling, and pyosalpinx. It is usually diagnosed by surgeons during diagnostic laparoscopy for other purposes. We present a rare case of isolated accessory tube torsion in a young adolescent. A 16-year-old virgin teen presented with a 24-hour history of aggravating right lower-quadrant abdominal pain and nausea without vomiting or fever. On examination she had right lower quadrant abdominal tenderness with no peritoneal signs. On pelvic ultrasound a right corpus luteum cyst was suspected, but right adnexal torsion could not be ruled out. On laparoscopy, torsion of the right accessory tube was diagnosed. Because of its ischemic and bluish appearance it was removed. The postoperative course was uneventful. Pathology confirmed the diagnosis. Torsion of an accessory fallopian tube is rare. An English literature search showed that the current case is the third reported overall and the first in a young adolescent. Because of the rarity of this congenital variation, and the low suspicion index for its existence, the diagnosis of accessory tube is often missed. Preventive removal of such findings should be considered but weighed against its possible surgical complications.


Simplified liquid-chromatographic assay of amiodarone and desethylamiodarone after solid-phase extraction.
We describe a rapid, simplified isocratic "high-performance" liquid-chromatographic method for simultaneous measurement of the antiarrhythmic drug amiodarone and its major metabolite, desethylamiodarone, in small volumes of sera (100 microL). Compared with liquid-liquid extraction, the solid-phase method of extraction saves time and glassware and improves reproducibility for small sample volumes. Amiodarone and desethylamiodarone could be measured at concentrations as low as 250 micrograms/L. Standard curves for the drug and metabolite are linear over the range of concentrations found in our patients. Within-run CVs (n = 6) ranged from 2.7% to 4.5% for amiodarone and from 4.0% to 5.7% for desethylamiodarone over the range of 250 to 4000 micrograms/L. Between-run CVs (n = 12) were 8.3% and 5.7% for amiodarone and desethylamiodarone, respectively. Commonly used cardiovascular medications do not interfere with the assay.


White-light cw-pumped supercontinuum generation in highly GeO(2)-doped-core photonic crystal fibers.
GeO(2)-doped-core photonic crystal fibers show greatly enhanced Kerr and Raman responses with respect to pure silica without any significant modification of the group-velocity dispersion curve. We show that such fibers allow a significant improvement of cw-pumped supercontinuum generation. We report for the first time to our knowledge the generation of a white-light cw-pumped supercontinuum spanning from 470 nm to more than 1750 nm.


Staging of multivessel percutaneous coronary interventions: an expert consensus statement from the Society for Cardiovascular Angiography and Interventions.
Percutaneous coronary interventions (PCIs) to treat multivessel coronary artery disease (MVCAD) may involve single-vessel or multivessel interventions, performed in one or more stages. This consensus statement reviews factors that may influence choice of strategy and includes six recommendations to guide decisions regarding staging of PCI. Every patient who undergoes PCI should receive optimal therapy for coronary disease, ideally before starting the procedure. Multivessel PCI at the time of diagnostic catheterization should be considered only if informed consent included the risks and benefits of multivessel PCI and the risks and benefits of alternative treatments. When considering multivessel PCI, the interventionist should develop a strategy regarding which stenoses to treat or evaluate, and their order, method, and timing. This strategy should maximize patient benefits, minimize patient risk, and consider the factors described in this article. For planned multivessel PCI, additional vessel(s) should be treated only if the first vessel is treated successfully and if anticipated contrast and radiation doses and patient and operator conditions are favorable. After the first stage of the planned multistage PCI, the need for subsequent PCI should be reviewed before it is performed. Third party payers and quality auditors should recognize that multistage PCI for MVCAD is neither an indication of poor quality nor an attempt to increase reimbursement when performed according to recommendations in this article.


Role of outer-pore residue Y380 in U-type inactivation of KV2.1 channels.
The interpretation of slow inactivation in potassium channels has been strongly influenced by work on C-type inactivation in Shaker channels. Slow inactivation in Shaker and some other potassium channels can be dramatically modulated by the state of the pore, including mutations at outer pore residue T449, which altered inactivation kinetics up to 100-fold. KV2.1, another voltage-dependent potassium channel, exhibits a biophysically distinct inactivation mechanism with a U-shaped voltage-dependence and preferential closed-state inactivation, termed U-type inactivation. However, it remains to be demonstrated whether U-type and C-type inactivation have different molecular mechanisms. This study examines mutations at Y380 (homologous to Shaker T449) to investigate whether C-type and U-type inactivation have distinct molecular mechanisms, and whether C-type inactivation can occur at all in KV2.1. Y380 mutants do not introduce C-type inactivation into KV2.1 and have little effect on U-type inactivation of KV2.1. Interestingly, two of the mutants tested exhibit twofold faster recovery from inactivation compared to wild-type channels. The observation that mutations have little effect suggests KV2.1 lacks C-type inactivation as it exists in Shaker and that C-type and U-type inactivation have different molecular mechanisms. Kinetic modeling predicts that all mutants inactivate preferentially, but not exclusively, from partially activated closed states. Therefore, KV2.1 exhibits a single U-type inactivation process including some inactivation from open as well as closed states.


Effect of pollination timing on the rate of apomictic reproduction revealed by RAPD markers in paspalum notatum.
Progeny tests employing molecular markers allow the identification of individuals originated by sexual means among the offspring of a facultative apomict. The objective of this work was to evaluate the effect of the pollination timing on the proportion of sexually formed individuals in progenies of a facultative apomictic Paspalum notatum genotype. Progeny families of approx. 30 plants each were generated at five different pollination times: 1-3 d pre-anthesis; at anthesis; and 2, 4 and 6 d post-anthesis. Cytoembryological analyses indicated that approx. 17% of the ovules carried a meiotic cytologically reduced embryo sac in florets formed simultaneously with those used for crosses. The parental plants and the five F1 families were analysed using RAPD molecular markers. Ninety-five oligonucleotides were assayed on the progenitors in order to search for male-specific bands. Eight primers presenting clear polymorphic bands were selected for use in the progeny tests. The proportion of sexually produced progeny reached 3.4% before anthesis and 20 % at anthesis, while pollination after anthesis generated only maternal plants. A second progeny of 97 plants obtained from pollination at anthesis produced 16 off-type plants (16.5%), of which only one was a B(III) hybrid (2n + n). Our results indicate that pollination at anthesis allows the greatest potential for sexuality to be expressed in this facultative apomictic genotype. When pollination is delayed as soon as 2 d after anthesis, only the aposporous sacs develop endosperm through pseudogamy to set seed.


Haematological findings in healthy and diseased rabbits, a multivariate analysis.
Haematological data from 94 diseased rabbits and 23 healthy adult rabbits were analysed statistically using principal components analysis and multiple discriminant function analysis. The 1st principal component accounted for 32% of the variation and was interpreted as showing total blood cellularity. This was decreased in the majority of diseased rabbits, principally due to anaemia, although in a few it was slightly increased possibly as a result of dehydration. The 2nd principal component (22% of the variation) had high positive loadings on monocyte and heterophil counts and weak negative loadings on the red cell characters. This component was interpreted as the response of the white blood cells to disease. When classification analysis was used to decide whether or not an animal was 'normal' the heterophil and lymphocyte counts alone provided almost as accurate an assessment as when all the blood parameters were taken into account. The role of these analyses in the diagnosis of disease is discussed.


Age and sex differences in coronal lower extremity alignment in a healthy Asian population.
The purpose of this study was to analyze the coronal alignment of lower extremities according to age and sex in a healthy population and demonstrate the differences. Standing full-lower limb anteroposterior (AP) radiographs of healthy volunteers (670 males and 782 females) aged 18-69 years were retrospectively analyzed. The hip-knee-ankle angle (HKA), lateral distal femoral angle (LDFA), medial proximal tibial angle (MPTA), joint line convergence angle and femoral bowing angle (FBA) were measured. The radiographic parameters were compared according to groups of age and sex. The proportion of volunteers with varus or valgus alignment more than 3° were also analyzed. With increasing age, HKA and LDFA varus increased. With increasing age, femoral medial bowing decreased. In addition, the HKA showed more varus alignment in males than in females (178.01° vs. 178.82°, P < 0.001). The MPTA was about 1° smaller in males than in females (P < 0.001). The proportion of patients with varus alignment of more than 3° increased with increasing age, with 16.9% in the 10-19 years old and 38.0% in the 60-69 years old groups. This study demonstrated that males showed more varus tibial alignments than females. Varus limb alignment, LDFA, and FBA also increases with age. In contrast, tibial alignment was constant across all age groups. Therefore, differences in lower extremity alignment according to age and sex should be considered in estimating individual prearthritic alignments.


Glucose-regulated protein 78 positivity as a predictor of poor survival in patients with renal cell carcinoma.
Glucose-regulated protein 78 (GRP78), a chaperone for newly formed proteins during folding and glycosylation, is associated with resistance to apoptosis in some forms of cancer. We assessed GRP78 expression and its correlation with clinicopathological parameters and survival. Immunohistochemistry was performed using formalin-fixed, paraffin-embedded specimens: 128 primary renal cell carcinoma (RCC) specimens (120 conventional and 8 other cell types) and 9 metastatic specimens. GRP78 positivity was determined based on intensity of staining and percentage of cells stained. Correlation of GRP78 positivity with clinicopathological parameters including patients' survival was evaluated. A statistically significant association was found between GRP78 positivity and higher tumor grade (G3; p <0.0001), advanced T stage (≥pT3; p = 0.0002), lymphovascular invasion (positive; p <0.0001), regional nodal involvement (≥N1; p = 0.0086), and distant metastases at presentation (M1; p = 0.001). Positivity of GRP78 expression was significantly associated with shorter disease-specific survival and shorter progression-free survival. Cox proportional hazard model showed that strong GRP78 positivity was an independent predictor of shortened progression-free survival in N0M0 RCC patients. There was a significant relationship between GRP78 expression levels and aggressiveness of RCC. Increased expression of GRP78 might be a useful parameter to predict shortened survival in patients with RCC.


Cucumber mosaic virus coat protein modulates the accumulation of 2b protein and antiviral silencing that causes symptom recovery in planta.
Shoot apical meristems (SAM) are resistant to most plant viruses due to RNA silencing, which is restrained by viral suppressors of RNA silencing (VSRs) to facilitate transient viral invasion of the SAM. In many cases chronic symptoms and long-term virus recovery occur, but the underlying mechanisms are poorly understood. Here, we found that wild-type Cucumber mosaic virus (CMVWT) invaded the SAM transiently, but was subsequently eliminated from the meristems. Unexpectedly, a CMV mutant, designated CMVRA that harbors an alanine substitution in the N-terminal arginine-rich region of the coat protein (CP) persistently invaded the SAM and resulted in visible reductions in apical dominance. Notably, the CMVWT virus elicited more potent antiviral silencing than CMVRA in newly emerging leaves of infected plants. However, both viruses caused severe symptoms with minimal antiviral silencing effects in the Arabidopsis mutants lacking host RNA-DEPENDENT RNA POLYMERASE 6 (RDR6) or SUPPRESSOR OF GENE SILENCING 3 (SGS3), indicating that CMVWT induced host RDR6/SGS3-dependent antiviral silencing. We also showed that reduced accumulation of the 2b protein is elicited in the CMVWT infection and consequently rescues potent antiviral RNA silencing. Indeed, co-infiltration assays showed that the suppression of posttranscriptional gene silencing mediated by 2b is more severely compromised by co-expression of CPWT than by CPRA. We further demonstrated that CPWT had high RNA binding activity leading to translation inhibition in wheat germ systems, and CPWT was associated with SGS3 into punctate granules in vivo. Thus, we propose that the RNAs bound and protected by CPWT possibly serve as templates of RDR6/SGS3 complexes for siRNA amplification. Together, these findings suggest that the CMV CP acts as a central hub that modulates antiviral silencing and VSRs activity, and mediates viral self-attenuation and long-term symptom recovery.


Refractory acquired haemophilia A in a patient with HIV treated with Emicizumab.
Acquired haemophilia is a rare coagulopathy characterized by acquired inhibitors directed against clotting factors resulting in bleeding episodes. A middle-aged woman with HIV developed refractory haemophilia with bleeding episodes resulting in recurrent hospitalizations despite several rounds of bypassing agents and several lines of immunosuppressive agents. She was eventually successfully treated with Emicizumab and has not had any major bleeding episodes for 3 years since initiation of this treatment. Emicizumab, which is a bispecific, FVIII-mimetic therapeutic antibody, has considerably reduced the annualized bleeding rates in congenital haemophiliacs with and without inhibitors and should be considered as an agent for acquired haemophilia to reduce recurrent bleeding episodes and even decrease inhibitor titer.


Experimental Outcrossing in Agaricus bisporus Revealed a Major and Unexpected Involvement of Airborne Mycelium Fragments.
Agaricus bisporus var. bisporus, the button mushroom, has a predominantly pseudohomothallic life cycle. Most of its spores are heterokaryotic and give rise to fertile heterokaryons. However, previous studies have suggested that outcrossing should not be rare in wild populations. In order to discover how outcrossing occurs, we experimentally favored it between aerial propagules of a fruiting donor mycelium and a delayed receiver mycelium that only invaded culture trays. Two donor/receiver pairs were studied, and potentially hybrid basidiomata collected on the receiver trays were analyzed with a mitochondrial marker, two unlinked nuclear CAPS markers, then haplotype markers based on DNA sequences obtained after PCR cloning of the rDNA ITS region and the fruk gene. For one of the two pairs, most basidiomata were hybrids between the donor and the receiver. Genotyping of the hybrids revealed only two genotypes consistent with outcrossing involving airborne mycelium fragments rather than basidiospores. The resident receiver heterokaryon that provided its mitochondria to the hybrid basidiomata is suspected to have had a trophic contribution to their growth and successful fruiting. The high level of heterozygosity and the cultivar introgression previously revealed in wild populations of this pseudohomothallic species may result from outcrossing involving airborne pieces of mycelium.


The Association between Sleep Problems, Sleep Medication Use, and Falls in Community-Dwelling Older Adults: Results from the Health and Retirement Study 2010.
Background. Very few studies have assessed the impact of poor sleep and sleep medication use on the risk of falls among community-dwelling older adults. The objective of this study was to evaluate the association between sleep problems, sleep medication use, and falls in community-dwelling older adults. Methods. The study population comprised a nationally representative sample of noninstitutionalized older adults participating in the 2010 Health and Retirement Study. Proportion of adults reporting sleep problems, sleep medication use, and fall was calculated. Multiple logistic regression models were constructed to examine the impact of sleep problems and sleep medication use on the risk of falls after controlling for covariates. Results. Among 9,843 community-dwelling older adults, 35.8% had reported a fall and 40.8% had reported sleep problems in the past two years. Sleep medication use was reported by 20.9% of the participants. Older adults who do have sleep problems and take sleep medications had a significant high risk of falls, compared to older adults who do not have sleep problems and do not take sleep medications. The other two groups also had significantly greater risk for falls. Conclusion. Sleep problems added to sleep medication use increase the risk of falls. Further prospective studies are needed to confirm these observed findings.


Regulation of contractile activity in vascular smooth muscle by protein kinases.
Control of the contraction/relaxation cycle in vascular smooth muscle is regulated by Ca2+ and the cyclic nucleotides, cAMP and cGMP. For the most part, the effectors of these intracellular messengers are the protein kinases. Four major protein kinases (myosin light chain kinase, protein kinase C, cAMP dependent protein kinase, and cGMP dependent protein kinase) have been identified in vascular smooth muscle. Substantial biochemical and physiological evidence exists supporting the involvement of Ca2+/calmodulin-mediated activation of myosin light chain kinase and phosphorylation of the 20,000 dalton P-light chain of myosin in the regulation of vascular contractile activity. However, alternative hypotheses exist which suggest that additional Ca2+ dependent regulatory mechanisms reside at other contractile protein sites. Calcium also activates protein kinase C, which requires phospholipid and diacylglycerol as co-factors instead of calmodulin. Protein kinase C also phosphorylates smooth muscle myosin P-light chain; however, phosphorylation occurs at a different site on the P-light chain and represses ATPase activity which has been stimulated by myosin light chain kinase-catalyzed phosphorylation. The precise physiological role of protein kinase C in modulating vascular smooth muscle contractile activity remains to be elucidated. Relaxation of vascular smooth muscle by some different relaxants is linked to either cAMP or cGMP formation. Correlative evidence also links activation of cAMP dependent protein kinase with relaxation. Two isozymes of cAMP dependent protein kinase exist in arterial smooth muscle; potential specific roles for each isozyme have not been elucidated. Mechanistically, relaxation mediated by both cyclic nucleotide-regulated protein kinases most likely involves primary effects on Ca2+ ion flux regulation rather than direct effects on contractile protein interactions. Activation of cGMP dependent protein kinase may be important in mediating the relaxant effects of endothelium derived relaxant factor or atrial natriuretic factor. Direct pharmacological modulation of smooth muscle vascular protein kinase activity represents an approach towards developing novel vasodilator agents. Various classes of agents, including phenothiazine antipsychotics, antidepressants, naphthalene sulfonamides, and certain lipophilic Ca2+ antagonists, inhibit myosin light chain kinase activity primarily by competition with the enzyme for Ca2+-calmodulin. However, additional inhibition via binding to the myosin P-light chain may also occur with some of these agents.(ABSTRACT TRUNCATED AT 400 WORDS)


Stable isotope analysis of saline water samples on a cavity ring-down spectroscopy instrument.
The analysis of the stable hydrogen and oxygen isotope composition of water using cavity ring-down spectroscopy (CRDS) instruments utilizing infrared absorption spectroscopy have been comprehensively tested. However, potential limitations of infrared spectroscopy for the analysis of highly saline water have not yet been evaluated. In this study, we assessed uncertainty arising from elevated salt concentrations in water analyzed on a CRDS instrument and the necessity of a correction procedure. We prepared various solutions of mixed salts and separate solutions with individual salts (NaCl, KCl, MgCl2, and CaCl2) using deionized water with a known stable isotope composition. Most of the individual salt and salt mixture solutions (some up to 340 g L(-1)) had δ-values within the range usual for CRDS analytical uncertainty (0.1‰ for δ (18)O and 1.0‰ for δ (2)H). Results were not compromised even when the total load of salt in the vaporizer reached ∼38.5 mg (equivalent to build up after running ∼100 ocean water samples). Therefore, highly saline mixtures can be successfully analyzed using CRDS, except highly concentrated MgCl2 solutions, without the need for an additional correction if the vaporizer is frequently cleaned and MgCl2 concentration in water is relatively low.


Potentially Important Contribution of Gas-Phase Oxidation of Naphthalene and Methylnaphthalene to Secondary Organic Aerosol during Haze Events in Beijing.
Naphthalene (Nap) and methylnaphthalene (MN) are the most abundant polycyclic aromatic hydrocarbons (PAHs) in atmosphere and have been proposed to be important precursors of anthropogenic secondary organic aerosol (SOA) derived from laboratory chamber experiments. In this study, atmospheric Nap/MN and their gas-phase photooxidation products were quantified by a Proton Transfer Reaction-Quadrupole interface Time-of-Flight Mass Spectrometer (PTR-QiTOF) during the 2016 winter in Beijing. Phthalic anhydride, a late generation product from Nap under high-NO x conditions, appeared to be more prominent than 2-formylcinnamaldehyde (early generation product), possibly due to more sufficient oxidation during the haze. 1,2-Phthalic acid (1,2-PhA), the hydrated form of phthalic anhydride, was capable of partitioning into aerosol phase and served as a tracer to explore the contribution of Nap to ambient SOA. The measured fraction in particle phase ( Fp) of 1,2-PhA averaged at 73 ± 13% with OA mass loadings of 52.5-87.8 μg/m3, lower than the value predicted by the absorptive partitioning model (100%). Using tracer product-based and precursor consumption-based methods, 2-ring PAHs (Nap and MN) were estimated to produce 14.9% (an upper limit) of the SOA formed in the afternoon during the wintertime haze, suggesting a comparable contribution of Nap and MN with monocyclic-aromatics on urban SOA formation.


Healthcare resource and lost labour costs of migraine headache in the US.
Migraine headache is responsible for significantly more healthcare resource and lost labour costs than previously reported. Costs associated with migraine were assessed via a survey conducted in 940 patients, 70% of whom responded. All met the International Headache Society's diagnostic criteria for migraine and had participated in one of two multicentre, single-dose, parallel-group, randomised, placebo-controlled clinical trials designed to assess the efficacy of an anti-migraine compound. Migraine frequency and costs, in terms of healthcare resource utilisation and lost labour (decreased productivity and missed workdays), were assessed. Over 90% of respondents visited a clinic and nearly 50% presented to an emergency room for treatment of migraine-related symptoms at least once in the year prior to the survey. These 648 respondents used an estimated $US529 199 per year in healthcare services. 89% of employed respondents reported that job performance was adversely affected by migraine and over 50% of them missed at least two days of work per month. Depending on the estimates used for migraine prevalence and using 1986 estimates of median earnings for the US work force, the extrapolated costs to employers ranged from $US5.6 billion to $US17.2 billion dollars annually due to decreased productivity and missed work days. The cost of migraine is not fully appreciated by the medical community or by society.


SmartStart: Results of a large point of entry study into preloading alcohol and associated behaviours.
There is a growing trend of preloading with alcohol before entering entertainment districts. It is claimed that this occurs to save money and that preloading may be a good indicator of harmful drinking and risk taking behaviours more generally. No study has collected data from a large sample as the participants entered entertainment districts and measured blood alcohol concentration (BAC) levels and self-reported drinking and risk taking behaviours in a systematic way. In this research, police and academics worked together to gauge the breadth and depth of preloading behaviours. In all, 3039 people completed a questionnaire and were breathalysed as they entered entertainment districts in Queensland, Australia. Of those, 2751 represented people from Brisbane and this data, collected every Thursday night to Sunday morning during the warm months, was analysed. More than 79% of people reported to preload and 71% returned a BAC greater than zero, both with little difference between the genders. Of preloaders, the mean BAC was 0.071, with 'to socialise with friends' being the primary reason given for preloading. Increasing preloading BAC was related to increasing risk taking and antisocial behaviours, as well as alcohol abuse and dependence. Older people entering entertainment districts had more accurate estimates of their BAC, yet 20% of our sample did not understand how the BAC system worked. Conducting the research was associated with a higher access rate to police and a lower arrest rate in the areas of data collection in comparison to the same nights 1year earlier. Preloading is widespread and involves moderate to heavy drinking in the Australian population visiting entertainment districts. Any interventions to curb drinking behaviours and reduce violence in night time entertainment districts need to involve approaches aimed at cultural phenomena, such as preloading behaviours.


Role of topical antibiotic ointment in the lateral graft following underlay myringoplasty: a prospective randomised study.
The objective of this study was to compare the graft outcome and postoperative infection of with and without the use of antibiotic ointment following myringoplasty for the treatment of chronic perforations. Randomized controlled trial. 135 chronic perforations were prospectively randomized to use of antibiotic ointment group (UAO, n = 68) or no use of antibiotic ointment group (NAO, n = 67) following myringoplasty. The graft outcomes and postoperative infection were compared among two groups at 6 months. At postoperative 6 months, the graft infection rate was 4.4% in the UAO group and 10.4% in the NAO group, the difference was not significant (P = 0.312).The graft success rates were 92.6% in the UAO group and 91.0% in the NAO group, the difference was not significant (P = 0.979). In the UAO group, 3 patients with purulence ear discharge resulted in a residual perforation although they received ofloxacin ear drops and intravenous antibiotic therapy treatment. In the NAO group, 6 patients with purulence ear discharge resulted in a residual perforation, only one, with mild purulence discharge was successfully treated and closed. In addition, no significant between-group differences were observed pre- (P = 0.746) or post- (P = 0.521) operative air bone gap (ABG) values or mean ABG gain (P = 0.745). However, granular myringitis with minimal moistness but without infection has been noted in 3 (4.4%) patients in the UAO group and in 5 (7.5%) in the NAO group, the difference was not significant (P = 0.699). Use and non-use of antibiotic ointments for lateral packing of graft are both comparable methods following myringoplasty for postoperative infection and graft outcomes.


Differential effects of melatonin on the stimulated release of LH from dispersed pituitary cells of the prepubertal female rat.
The effect of melatonin on the stimulated release of LH from prepubertal female rat pituitary cells in vitro was investigated. Significant inhibition of LH-releasing hormone and calcium ionophore-induced LH release was seen but not of potassium-induced release. These results suggest a specific interaction between melatonin and the endogenous events leading to LH release, and may implicate melatonin as an important neuroendocrine component of pubertal development in this species.


Diversity of Secondary Metabolism in Aspergillus nidulans Clinical Isolates.
The filamentous fungus Aspergillus nidulans has been a primary workhorse used to understand fungal genetics. Much of this work has focused on elucidating the genetics of biosynthetic gene clusters (BGCs) and the secondary metabolites (SMs) they produce. SMs are both niche defining in fungi and of great economic importance to humans. Despite the focus on A. nidulans, very little is known about the natural diversity in secondary metabolism within this species. We determined the BGC content and looked for evolutionary patterns in BGCs from whole-genome sequences of two clinical isolates and the A4 reference genome of A. nidulans Differences in BGC content were used to explain SM profiles determined using liquid chromatography-high-resolution mass spectrometry. We found that in addition to genetic variation of BGCs contained by all isolates, nine BGCs varied by presence/absence. We discovered the viridicatumtoxin BGC in A. nidulans and suggest that this BGC has undergone a horizontal gene transfer from the Aspergillus section Nigri lineage into Penicillium sometime after the sections Nigri and Nidulantes diverged. We identified the production of viridicatumtoxin and several other compounds previously not known to be produced by A. nidulans One isolate showed a lack of sterigmatocystin production even though it contained an apparently intact sterigmatocystin BGC, raising questions about other genes and processes known to regulate this BGC. Altogether, our work uncovers a large degree of intraspecies diversity in BGC and SM production in this genetic model species and offers new avenues to understand the evolution and regulation of secondary metabolism.IMPORTANCE Much of what we know about the genetics underlying secondary metabolite (SM) production and the function of SMs in the model fungus Aspergillus nidulans comes from a single reference genome. A growing body of research indicates the importance of biosynthetic gene cluster (BGC) and SM diversity within a species. However, there is no information about the natural diversity of secondary metabolism in A. nidulans We discovered six novel clusters that contribute to the considerable variation in both BGC content and SM production within A. nidulans We characterize a diverse set of mutations and emphasize how findings of single nucleotide polymorphisms (SNPs), deletions, and differences in evolutionary history encompass much of the variation observed in nonmodel systems. Our results emphasize that A. nidulans may also be a strong model to use within-species diversity to elucidate regulatory cross talk, fungal ecology, and drug discovery systems.


A novel approach to label-free sensing: diffractive optics technology (dot).
Label-free detection methods have played a very significant role in drug design and refinement. They have been used primarily during secondary screening and for in-depth characterization of biomolecular interactions. Misconceptions about the accessibility of these platforms, since they often require specialized training, throughput and robustness in complex media have hampered their adoption in the earliest phases of discovery not to mention their significant and unrealized potential in qualifying reagents for high throughput screening or during novel assay development. A new wave of more cost effective, robust and accessible platforms has made significant inroads, demonstrating that significant information can be derived from these methods all along the drug discovery research continuum. One of these recent entrants, the dotLab System uses diffractive optics technology (dot) to detect biomolecular interactions and can be used for a wide variety of applications in the study of a broad spectrum of biological analytes including proteins, DNA and even microorganisms.


A comparative study of rank aggregation methods for partial and top ranked lists in genomic applications.
Rank aggregation (RA), the process of combining multiple ranked lists into a single ranking, has played an important role in integrating information from individual genomic studies that address the same biological question. In previous research, attention has been focused on aggregating full lists. However, partial and/or top ranked lists are prevalent because of the great heterogeneity of genomic studies and limited resources for follow-up investigation. To be able to handle such lists, some ad hoc adjustments have been suggested in the past, but how RA methods perform on them (after the adjustments) has never been fully evaluated. In this article, a systematic framework is proposed to define different situations that may occur based on the nature of individually ranked lists. A comprehensive simulation study is conducted to examine the performance characteristics of a collection of existing RA methods that are suitable for genomic applications under various settings simulated to mimic practical situations. A non-small cell lung cancer data example is provided for further comparison. Based on our numerical results, general guidelines about which methods perform the best/worst, and under what conditions, are provided. Also, we discuss key factors that substantially affect the performance of the different methods.


Establishment of stable cell line for inducing KAP1 protein expression.
Generation of the stable cell lines is a highly efficient tool in functional studies of certain genes or proteins, where the particular genes or proteins are inducibly expressed. The KRAB-associated protein-1 (KAP1) is an important transcription regulatory protein, which is investigated in several molecular biological studies. In this study, we have aimed to generate a stable cell line for inducing KAP1 expression. The recombinant plasmid pcDNA5/FRT/TO-KAP1 was constructed at first, which was then transfected into Flp-In™T-REx™-HEK293 cells to establish an inducible pcDNA5/FRT/TO-KAP1-HEK293 cell line. The Western blot analysis showed that the protein level of KAP1 is over-expressed in the established stable cell line by doxycycline induction, both dose and time dependently. Thus we have successfully established stable pcDNA5/FRT/TO-KAP1-HEK293 cell line, which can express KAP1 inducibly. This inducible cell line might be very useful for KAP1 functional studies.


Inferring bacterial genome flux while considering truncated genes.
Bacterial gene content variation during the course of evolution has been widely acknowledged and its pattern has been actively modeled in recent years. Gene truncation or gene pseudogenization also plays an important role in shaping bacterial genome content. Truncated genes could also arise from small-scale lateral gene transfer events. Unfortunately, the information of truncated genes has not been considered in any existing mathematical models on gene content variation. In this study, we developed a model to incorporate truncated genes. Maximum-likelihood estimates (MLEs) of the new model reveal fast rates of gene insertions/deletions on recent branches, suggesting a fast turnover of many recently transferred genes. The estimates also suggest that many truncated genes are in the process of being eliminated from the genome. Furthermore, we demonstrate that the ignorance of truncated genes in the estimation does not lead to a systematic bias but rather has a more complicated effect. Analysis using the new model not only provides more accurate estimates on gene gains/losses (or insertions/deletions), but also reduces any concern of a systematic bias from applying simplified models to bacterial genome evolution. Although not a primary purpose, the model incorporating truncated genes could be potentially used for phylogeny reconstruction using gene family content.


Escherichia coli Adhesion and Biofilm Formation on Polydimethylsiloxane are Independent of Substrate Stiffness.
Bacterial adhesion and biofilm formation on the surface of biomedical devices are detrimental processes that compromise patient safety and material functionality. Several physicochemical factors are involved in biofilm growth, including the surface properties. Among these, material stiffness has recently been suggested to influence microbial adhesion and biofilm growth in a variety of polymers and hydrogels. However, no clear consensus exists about the role of material stiffness in biofilm initiation and whether very compliant substrates are deleterious to bacterial cell adhesion. Here, by systematically tuning substrate topography and stiffness while keeping the surface free energy of polydimethylsiloxane substrates constant, we show that topographical patterns at the micron and submicron scale impart unique properties to the surface which are independent of the material stiffness. The current work provides a better understanding of the role of material stiffness in bacterial physiology and may constitute a cost-effective and simple strategy to reduce bacterial attachment and biofilm growth even in very compliant and hydrophobic polymers.


Peer-to-Patient-Aligned Care Team (Peer-to-PACT; P2P), a Peer-Led Home Visit Intervention Program for Targeting and Improving Long-term Care Services and Support for Veterans With High Needs and High Risk: Protocol for a Mixed Methods Feasibility Study.
Keeping older veterans with high needs and high risk (HNHR) who are at risk of long-term institutional care safely in their homes for as long as possible is a Department of Veterans Affairs priority. Older veterans with HNHR face disproportionate barriers and disparities to engaging in their care, including accessing care and services. Veterans with HNHR often have poor ability to maintain health owing to complicated unmet health and social needs. The use of peer support specialists (peers) is a promising approach to improving patient engagement and addressing unmet needs. The Peer-to-Patient-Aligned Care Team (Peer-to-PACT; P2P) intervention is a multicomponential home visit intervention designed to support older veterans with HNHR to age in place. Participants receive a peer-led home visit to identify unmet needs and home safety risks aligned with the age-friendly health system model; care coordination, health care system navigation, and linking to needed services and resources in collaboration with their PACT; and patient empowerment and coaching using Department of Veterans Affairs whole health principles. The primary aim of this study is to evaluate the preliminary effect of the P2P intervention on patient health care engagement. The second aim is to identify the number and types of needs and unmet needs as well as needs addressed using the P2P needs identification tool. The third aim is to evaluate the feasibility and acceptability of the P2P intervention delivered over 6 months. We will use a quantitative-qualitative convergent mixed methods approach to evaluate the P2P intervention outcomes. For our primary outcome, we will conduct an independent, 2-tailed, 2-sample t test to compare the means of the 6-month pre-post differences in the number of outpatient PACT encounters between the intervention and matched comparison groups. Qualitative data analysis will follow a structured rapid approach using deductive coding as well as the Consolidated Framework for Implementation Research. Study enrollment began in July 2020 and was completed in March 2022. Our sample size consists of 114 veterans: 38 (33.3%) P2P intervention participants and 76 (66.7%) matched comparison group participants. Study findings are expected to be published in late 2023. Peers may help bridge the gap between PACT providers and veterans with HNHR by evaluating veterans' needs outside of the clinic, summarizing identified unmet needs, and developing team-based solutions in partnership with the PACT. The home visit component of the intervention provides eyes in the home and may be a promising and innovative tool to improve patient engagement. DERR1-10.2196/46156.


Knocking Out SST Gene of BGC823 Gastric Cancer Cell by CRISPR/Cas9 Enhances Migration, Invasion and Expression of SEMA5A and KLF2.
The impact and potential molecular mechanisms of SST in the occurrence and development of GC have not been determined. Two pairs of sgRNA and reporter were designed according to targeting sequence of SST gene for double-nicking. Plasmids were transfected into 293T for selecting sgRNA with higher cutting efficiency. The subline which has knocked-out SST gene were selected by FACS and verified by sequencing and expression level. Moreover, the migration and invasion ability was evaluated by wound healing and transwell after knocking out SST. Besides, the protein expression of SEMA5A and KLF2 were observed by Western blotting and LSCM. Last, we detected the expression levels of SST, SEMA5A, and KLF2 in GC tissues by Western blotting. The results revealed that the new subline 1E9, which had knocked out SST gene, was established by CRISPR/Cas9. In addition, the knockout of SST in GC cells markedly increased migration and invasion ability. The results also demonstrated that the knockout of SST increased the expression of SEMA5A and KLF2. The expression level of SST was decreased in GC tissues, and its decrease was associated with overexpression of SEMA5A and KLF2. SST plays an inhibitory role in the migration and invasion of GC cell BGC823. The protein expression levels of SEMA5A and KLF2 were enhanced in GC cells and tissues lacking SST expression.


Advanced pneumatic compression for treatment of lymphedema of the head and neck: a randomized wait-list controlled trial.
Lymphedema associated with head and neck cancer (HNC) therapy causes adverse clinical outcomes. Standard treatment includes professionally administered complete decongestive therapy (CDT). Cost and availability of trained therapists are known barriers to therapy. Advanced pneumatic compression devices (APCD) may address these issues. A randomized, wait-list controlled trial was undertaken to evaluate an APCD in post-treatment HNC patients with lymphedema. Eligible patients had completed treatment for HNC, were disease free, and had lymphedema at enrollment. Participants were randomized to wait-list lymphedema self-management (standard of care) or lymphedema self-management plus the use of the APCD bid. Safety (CTCAE V4.0) and feasibility were primary endpoints; secondary endpoints included efficacy measure by objective examination and patient reported outcomes (symptoms, quality of life, function), adherence barriers, and satisfaction. Assessments were conducted at baseline and weeks 4 and 8. Forty-nine patients were enrolled (wait-list n = 25; intervention n = 24). In total, forty-three patients completed the study. No device-related Serious Adverse Events were reported. Most patients used the APCD once per day, instead of the prescribed twice per day, citing time related factors as barriers to use. APCD use was associated with significant improvement in perceived ability to control lymphedema (p = 0.003) and visible external swelling (front view p < 0.001, right view p = 0.004, left p = 0.005), as well as less reported pain. This trial supports the safety and feasibility of the APCD for the treatment of secondary lymphedema in head and neck cancer patients. In addition, preliminary data supports efficacy.


[Induction of antitumor immune response in the mouse model after vaccination with B7.1 expressing tumor cells].
The aim of the study was the induction of an antitumor immune response by genetic modification of tumor cells. This was done by transfecting the costimulatory molecule B7.1 into a murine tumor cell line SCCVII/SF in order to increase T cell recognition and to install an immunologic memory. One cohort of immunocompetent mice C3 H/HeN were injected with B7.1 expressing tumor cells, while the control group received parental B7.1 negative tumor cells. In a second step those immunized mice were rechallenged by parental tumor cells and tumor growth was compared to a new control group. Transfection with B7.1 prohibits outgrowth of the SCCVII cell line. Animals that have been vaccinated in this way are partially immune towards a secondary exposition to B7.1 negative tumor cells. Outgrowth of this recurrent tumor is slowed down. Such an immunization builds up an immunologic memory. Vaccination with B7.1 expressing tumor cells lead to a partial protective tumor immunity in the SCCVII-C3 H/HeN mouse model.


Insertion of a Yop translocation pore into the macrophage plasma membrane by Yersinia enterocolitica: requirement for translocators YopB and YopD, but not LcrG.
The Yersinia survival strategy is based on its ability to inject effector Yops into the cytosol of host cells. Translocation of these effectors across the eukaryotic cell membrane requires YopB, YopD and LcrG, but the mechanism is unclear. An effector polymutant of Y. pseudotuberculosis has a YopB-dependent contact haemolytic activity, indicating that YopB participates in the formation of a pore in the cell membrane. Here, we have investigated the formation of such a pore in the plasma membrane of macrophages. Infection of PU5-1.8 macrophages with an effector polymutant Y. enterocolitica led to complete flattening of the cells, similar to treatment with the pore-forming streptolysin O from Streptococcus pyogenes. Upon infection, cells released the low-molecular-weight marker BCECF (623 Da) but not the high-molecular-weight lactate dehydrogenase, indicating that there was no membrane lysis but, rather, insertion of a pore of small size into the macrophage plasma membrane. Permeation to lucifer yellow CH (443 Da) but not to Texas red-X phalloidin (1490 Da) supported this hypothesis. All these events were found to be dependent not only on translocator YopB as expected but also on YopD, which was required equally. In contrast, LcrG was not necessary. Consistently, lysis of sheep erythrocytes was also dependent on YopB and YopD, but not on LcrG.


Bilateral extensive emphysematous pyelonephritis. A case report.
We report a case involving a patient with bi- lateral emphysematous pyelonephritis (EPN) who was successfully treated with antibiotics and nephrostomy sparing nephrectomy. A 54-year-old diabetic man presented with the chief complaint of progressive abdominal pain for 10 days. Blood tests indicated acute renal failure and hyperglycemic hyperosmolar state. An abdominal radiograph disclosed gas distributed over regions where the bilateral kidneys are located and the right para-lumbosacral area. Computed tomography revealed bilateral enlarged kidneys with the presence of gas in the renal parenchyma and renal pelvis, left peri-renal space, bilateral ureter, bladder, prostate gland and right scrotum. Cultures of urine and blood grew Escherichia coli. The patient was successfully treated with antibiotics coupled with nephrostomy. Timely recognition and initiation of effective antibiotic therapy and nephrostomy may cure the patient with bilateral EPN, while sparing nephrectomy. Regarding clinical manifestations, EPN is nonspecific; hence, radiographs should be taken for diabetic patients with abdominal pain (especially those with poorly controlled blood sugar) to enable early diagnosis of potential EPN.


A peptide hydrogel derived from a fragment of human cardiac troponin C.
The human cardiac troponin C peptide fragment H-V(9)EQLTEEQKNEFKAAFDIFVLGA(31)-OH, which covers helix-A in the native protein, self-assembles into β-sheet fibrils in solution. These fibrils further entangle to give a hydrogel. This peptide may therefore serve as a template for development of novel biomaterials.


Solitary amyloid tumor of the tongue base.
The purpose of this article is to present a rare case of localized, solitary amyloid tumor of tongue base and emphasize some of the characteristic features of challenging clinical and histopathologic diagnosis. In this paper, we focused on the clinical and pathological specifications of this rare tumor, so any unnecessary examinations or measures may be spared. Negative staining of amyloid material with AAC and osseous metaplasia noted in the histopathologic examination may not be thought as definite criteria for localized amyloidosis, but a supporter of localized, solitary amyloid tumor diagnosis.


Effect of noise correlation on detectability of disk signals in medical imaging.
Pixel signal-to-noise ratio is one accepted measure of image quality for predicting observer performance in medical imaging. We have found, however, that images with equal pixel signal-to-noise ratio (SNRp) but different correlation properties give quite different observer-performance measures for a simple detection experiment. The SNR at the output of an ideal detector with the ability to prewhiten the noise is also a poor predictor of human performance for disk signals in high-pass noise. We have found constant observer efficiencies for humans relative to the performance of a nonprewhitening detector for this task.


Surgical management of subarachnoid hemorrhage.
Spontaneous subarachnoid hemorrhage is usually caused by a ruptured cerebral aneurysm. Aneurysmal rupture classically presents with sudden severe headache, often accompanied by an altered mental status. Diagnosis is made with computed tomography or lumbar puncture. Patients with ruptured cerebral aneurysms are at risk for rebleeding, cerebral artery vasospasm (and subsequent ischemia or stroke), and hydrocephalus. Early surgical clipping of the aneurysm under the microscope is usually the initial treatment of choice. This surgery prevents rebleeding and allows for safe use of pressors in the event that clinical vasospasm develops. Factors that would favor delayed surgery, "coiling" procedures, or conservative management include poor patient condition, basilar artery aneurysms, and unusually large or irregular aneurysms. Patients with ruptured aneurysms are treated with nimodipine, a calcium-channel blocker, to help prevent vasospasm-related ischemia. The degree of vasospasm that develops in the first 2 wks after aneurysmal rupture is assessed by transcranial Doppler sonography and cerebral angiography, in addition to the clinical examination. Patients with symptomatic vasospasm are kept well hydrated and treated with pressors (provided the aneurysm has been successfully clipped).


Big five personality and adolescent Internet addiction: The mediating role of coping style.
This study examined the unique associations between big five personality traits and adolescent Internet addiction (IA), as well as the mediating role of coping style underlying these relations. Our theoretical model was tested with 998 adolescents. Participants provided self-report data on demographic variables, big five personality traits, coping style, and IA. After controlling for demographic variables, it was found that agreeableness and conscientiousness were negatively associated with IA, whereas extraversion, neuroticism, and openness to experience were positively associated with IA. Mediation analyses further indicated that conscientiousness had an indirect impact on adolescent IA through decreased emotion-focused coping, whereas extraversion, neuroticism, openness to experience had indirect impacts on adolescent IA through increased emotion-focused coping. In contrast, problem-focused coping had no mediating role. These findings suggest that emotion-focused coping may, in part, account for the association between big five personality and adolescent IA.


A score system for complete cytoreduction in selected recurrent ovarian cancer patients undergoing secondary cytoreductive surgery: predictors- and nomogram-based analyses.
To test the applicability of the Arbeitsgemeinschaft Gynäkologische Onkologie (AGO) and Memorial Sloan Kettering (MSK) criteria in predicting complete cytoreduction (CC) in patients undergoing secondary cytoreductive surgery (SCS) for recurrent ovarian cancer (ROC). Data of consecutive patients undergoing SCS were reviewed. The Arbeitsgemeinschaft Gynäkologische Onkologie OVARian cancer study group (AGO-OVAR) and MSK criteria were retrospectively applied. Nomograms, based on AGO criteria, MSK criteria and both AGO and MSK criteria were built in order to assess the probability to achieve CC at SCS. Overall, 194 patients met the inclusion criteria. CC was achieved in 161 (82.9%) patients. According to the AGO-OVAR criteria, we observed that CC was achieved in 87.0% of patients with positive AGO score. However, 45 out of 71 (63.4%) patients who did not fulfilled the AGO score had CC. Similarly, CC was achieved in 87.1%, 61.9% and 66.7% of patients for whom SCS was recommended, had to be considered and was not recommended, respectively. In order to evaluate the predictive value of the AGO-OVAR and MSK criteria we built 2 separate nomograms (c-index: 0.5900 and 0.5989, respectively) to test the probability to achieve CC at SCS. Additionally, we built a nomogram using both the aforementioned criteria (c-index: 0.5857). The AGO and MSK criteria help identifying patients deserving SCS. However, these criteria might be strict, thus prohibiting a beneficial treatment in patients who do not met these criteria. Further studies are needed to clarify factors predicting CC at SCS.


UV and genotoxic stress induce ATR relocalization in mouse spermatocytes.
During meiosis, phosphorylation of H2AX is one of the earliest cellular responses to the generation of DNA double-strand breaks (DSBs) by the SPO11 topoisomerase. ATM is the kinase which mediates the formation of phosphorylated H2AX (H2AX) meiotic foci, while ATR is the kinase which signals chromosome asynapsis at the level of the XY bivalent. To investigate the possible role of ATR also in DNA damage signalling in meiotic cells, we studied the effect of UV radiation and chemotherapy drugs on H2AX phosphorylation and ATR relocalization in mouse pachytene spermatocytes. Here, we report that UV, a single strand break DNA-damaging agent, induces ATR relocalization from the XY sex body to nuclear foci and intense H2AX phosphorylation. Other DNA damage proteins such as MDC1, NBS1 and 53BP1 showed a similar relocalization following UVA microirradiation of spermatocytes. We found that DNA damage induced by UV increased the intensity and the number of H2AX foci also in Atm null spermatocytes. Inhibition of RNA synthesis was found to induce the formation of H2AX foci, but it did not influence the DNA damage response to UV irradiation. Finally, exposure of spermatocytes to double strand break DNA-damaging agents such as cisplatin, bleomycin or etoposide also induced ATR relocalization and intense H2AX phosphorylation and led to anomalies in synaptonemal assembly. Our results demonstrate that DNA damage induced by genotoxic stress can activate ATR and influence meiotic chromatin remodelling through H2AX phosphorylation, likely as part of a response which normally ensures germ cell genomic integrity.


Bioorthogonal Elastin-like Polypeptide Scaffolds for Immunoassay Enhancement.
Artificial multiprotein complexes are sought after reagents for biomolecular engineering. A current limiting factor is the paucity of molecular scaffolds which allow for site-specific multicomponent assembly. Here, we address this limitation by synthesizing bioorthogonal elastin-like polypeptide (ELP) scaffolds containing periodic noncanonical l-azidohomoalanine amino acids in the guest residue position. The nine azide ELP guest residues served as conjugation sites for site-specific modification with dibenzocyclooctyne (DBCO)-functionalized single-domain antibodies (SdAbs) through strain-promoted alkyne-azide cycloaddition (SPAAC). Sortase A and ybbR tags at the C- and N-termini of the ELP scaffold provided two additional sites for derivatization with small molecules and peptides by Sortase A and 4'-phosphopantetheinyl transferase (Sfp), respectively. These functional groups are chemically bioorthogonal, mutually compatible, and highly efficient, thereby enabling synthesis of multi-antibody ELP complexes in a one-pot reaction. We demonstrate application of this material for enhancing the performance of sandwich immunoassays of the recombinant protein mCherry. In undiluted human plasma, surfaces modified with multi-antibody ELP complexes showed between 2.3- and 14.3-fold improvement in sensitivity and ∼30-40% lower limits of detection as compared with nonspecifically adsorbed antibodies. Dual-labeled multi-antibody ELP complexes were further used for cytometric labeling and analysis of live eukaryotic cells. These results demonstrate how multiple antibodies complexed onto bioorthogonal protein-based polymers can be used to enhance immunospecific binding interactions through multivalency effects.


Genomic distribution and inter-sample variation of non-CpG methylation across human cell types.
DNA methylation plays an important role in development and disease. The primary sites of DNA methylation in vertebrates are cytosines in the CpG dinucleotide context, which account for roughly three quarters of the total DNA methylation content in human and mouse cells. While the genomic distribution, inter-individual stability, and functional role of CpG methylation are reasonably well understood, little is known about DNA methylation targeting CpA, CpT, and CpC (non-CpG) dinucleotides. Here we report a comprehensive analysis of non-CpG methylation in 76 genome-scale DNA methylation maps across pluripotent and differentiated human cell types. We confirm non-CpG methylation to be predominantly present in pluripotent cell types and observe a decrease upon differentiation and near complete absence in various somatic cell types. Although no function has been assigned to it in pluripotency, our data highlight that non-CpG methylation patterns reappear upon iPS cell reprogramming. Intriguingly, the patterns are highly variable and show little conservation between different pluripotent cell lines. We find a strong correlation of non-CpG methylation and DNMT3 expression levels while showing statistical independence of non-CpG methylation from pluripotency associated gene expression. In line with these findings, we show that knockdown of DNMTA and DNMT3B in hESCs results in a global reduction of non-CpG methylation. Finally, non-CpG methylation appears to be spatially correlated with CpG methylation. In summary these results contribute further to our understanding of cytosine methylation patterns in human cells using a large representative sample set.


Functional changes induced by necrotic laryngitis in double muscled calves.
The effects of necrotic laryngitis on the mechanics of breathing and gas exchange were investigated in five Belgian blue double muscled calves two to three months old. All the animals showed the typical clinical picture of the respiratory syndrome associated with naturally occurring necrotic laryngitis. Highly significant increases in total pulmonary resistance, minute viscous work of breathing and alveolar-arterial oxygen gradient, and highly significant decreases in dynamic lung compliance and arterial oxygen tension were recorded in the infected animals, when compared to reference values for healthy cattle. The ratio of inspiratory to expiratory viscous work of breathing was also significantly increased probably because of a partial collapse of the extrathoracic trachea during inspiration. It was concluded that necrotic laryngitis disturbs pulmonary function to such an extent that it impedes the growing process and predisposes the infected animals to secondary bronchopneumonia and ventilatory failure due to respiratory muscle fatigue.


Complete genomic sequence and comparative analysis of the tumorigenic poxvirus Yaba monkey tumor virus.
The Yatapoxvirus genus of poxviruses is comprised of Yaba monkey tumor virus (YMTV), Tanapox virus, and Yaba-like disease virus (YLDV), which all have the ability to infect primates, including humans. Unlike other poxviruses, YMTV induces formation of focalized histiocytomas upon infection. To gain a greater understanding of the Yatapoxvirus genus and the unique tumor formation properties of YMTV, we sequenced the 134,721-bp genome of YMTV. The genome of YMTV encodes at least 140 open reading frames, all of which are also found as orthologs in the closely related YLDV. However, 13 open reading frames found in YLDV are completely absent from YMTV. Common to both YLDV and YMTV are the unusually large noncoding regions between many open reading frames. To determine whether any of these noncoding regions might be functionally significant, we carried out a comparative analysis between the putative noncoding regions of YMTV and similar noncoding regions from other poxviruses. This approach identified three new gene poxvirus families, defined as orthologs of YMTV23.5L, YMTV28.5L, and YMTV120.5L, which are highly conserved in virtually all poxvirus species. Furthermore, the comparative analysis also revealed a 40-bp nucleotide sequence at approximately 14,700 bases from the left terminus that was 100% identical in the comparable intergene site within members of the Yatapoxvirus, Suipoxvirus, and Capripoxvirus genera and 95% conserved in the Leporipoxvirus genus. This conserved sequence was shown to function as a poxvirus late promoter element in transfected and infected cells, but other functions, such as an involvement in viral replication or packaging, cannot be excluded. Finally, we summarize the predicted immunomodulatory protein repertoire in the Yatapoxvirus genus as a whole.


Cloning, tissue expression, and chromosomal location of the mouse insulin receptor substrate 4 gene.
The insulin receptor substrates (IRSs) are key proteins in signal transduction from the insulin receptor. Recently, we discovered a fourth member of this family, designated IRS-4, cloned its complementary DNA from the human embryonic kidney 293 cell line, and characterized its signaling properties in this cell line. As part of an investigation of the physiological role of this IRS, we have now cloned the mouse IRS-4 gene and determined its tissue expression and chromosomal location. The coding region of the mouse IRS-4 gene contains no introns, and in this regard is the same as that of the genes for IRS-1 and -2. The predicted amino acid sequence of mouse IRS-4 is highly homologous with that of human IRS-4; the pleckstrin homology domain, the phosphotyrosine-binding domain, and the tyrosine phosphorylation motifs are especially well conserved. The tissue distribution of IRS-4 in the mouse was determined by analysis for the expression of its messenger RNA by RT-PCR and for the protein itself by immunoprecipitation and immunoblotting. The messenger RNA was detected in skeletal muscle, brain, heart, kidney, and liver, but the protein itself was not detected in any tissue. These results indicate that IRS-4 is a very rare protein. The chromosomal locations of the mouse IRS-4 and IRS-3 genes were determined by interspecific back-cross analysis and were found to be on chromosomes X and 5, respectively. As the mouse genes for IRS-1 and -2 are on chromosomes 1 and 8, respectively, each IRS gene resides on a different chromosome.


Photochemical production of singlet oxygen from particulate organic matter.
Dissolved organic matter is established as one of the most relevant photosensitizers in aquatic environments, producing singlet oxygen (1O2) alongside other photochemically produced reactive intermediates. While the production of 1O2 from DOM has been well studied, the relative importance of particulate organic matter (POM) to the overall 1O2 production is less well understood. POM is known to play an important role in pollutant fate through the sorption and transport of hydrophobic pollutants. If POM is directly involved in 1O2 production, sorbed molecules would be expected to undergo enhanced photodegradation. In this work, synthetic POM was prepared by coating silica particles with commercial humic acid. The photochemical behavior of these POM samples was compared to dissolved commercial humic acids (DOM). Suspended natural sediment was also studied to test the environmental relevance of the synthetic POM model. Synthetic POM particles appear to simulate well the 1O2-production of suspended sediment. The 1O2 concentrations experienced by POM-sorbed probe molecules was up to 30% higher than experienced by DOM-sorbed ones, even though the aqueous concentration of 1O2 in irradiated POM suspensions was much lower than the analogous DOM solutions. These results were interpreted with a reaction-diffusion model, which suggested that the production rate of 1O2 by POM is lower than DOM, but the loss of 1O2 from the POM-phase is also lower than DOM. Based on the experimental results of this study, calculations were conducted to estimate the impact of removing POM on 1O2-mediated processes. These calculations indicate that compounds with a log Koc value near 4 will be most affected by removal of POM and that the magnitude of the effect is proportional to the fraction of the total organic matter represented by POM. This study demonstrates that particles can play an important role in the degradation of organic compounds via aquatic photochemistry.


Human papillomavirus infection of the lower female genital tract: association with multicentric neoplasia.
Fifty-four patients with multiple condylomatous lesions and/or multicentric neoplasia were identified in a consecutive series of 2,627 colposcopic examinations (2%). Forty-one percent presented with condylomata only and 57% exhibited associated intraepithelial or invasive neoplasia in at least one site. Only one patient was found to have multicentric neoplasia with no condylomatous features. There was a significant increase in mean age between patients presenting with multiple condylomatous lesions and those who had intraepithelial neoplasia as well. An association with pregnancy was found in 17% of the cases, and with renal transplant in 5%. Virus particles compatible with human papillomavirus were found in 62% of the cases examined. Evidence for a causative role in the development of neoplasia of the lower female genital tract is considered.


An Analysis of Android Malware Classification Services.
The increasing number of Android malware forced antivirus (AV) companies to rely on automated classification techniques to determine the family and class of suspicious samples. The research community relies heavily on such labels to carry out prevalence studies of the threat ecosystem and to build datasets that are used to validate and benchmark novel detection and classification methods. In this work, we carry out an extensive study of the Android malware ecosystem by surveying white papers and reports from 6 key players in the industry, as well as 81 papers from 8 top security conferences, to understand how malware datasets are used by both. We, then, explore the limitations associated with the use of available malware classification services, namely VirusTotal (VT) engines, for determining the family of an Android sample. Using a dataset of 2.47 M Android malware samples, we find that the detection coverage of VT's AVs is generally very low, that the percentage of samples flagged by any 2 AV engines does not go beyond 52%, and that common families between any pair of AV engines is at best 29%. We rely on clustering to determine the extent to which different AV engine pairs agree upon which samples belong to the same family (regardless of the actual family name) and find that there are discrepancies that can introduce noise in automatic label unification schemes. We also observe the usage of generic labels and inconsistencies within the labels of top AV engines, suggesting that their efforts are directed towards accurate detection rather than classification. Our results contribute to a better understanding of the limitations of using Android malware family labels as supplied by common AV engines.


[Qualitative analysis: theory, steps and reliability].
This essay seeks to conduct in-depth analysis of qualitative research, based on benchmark authors and the author's own experience. The hypothesis is that in order for an analysis to be considered reliable, it needs to be based on structuring terms of qualitative research, namely the verbs 'comprehend' and 'interpret', and the nouns 'experience', 'common sense' and 'social action'. The 10 steps begin with the construction of the scientific object by its inclusion on the national and international agenda; the development of tools that make the theoretical concepts tangible; conducting field work that involves the researcher empathetically with the participants in the use of various techniques and approaches, making it possible to build relationships, observations and a narrative with perspective. Finally, the author deals with the analysis proper, showing how the object, which has already been studied in all the previous steps, should become a second-order construct, in which the logic of the actors in their diversity and not merely their speech predominates. The final report must be a theoretic, contextual, concise and clear narrative.


Improved outcome in haemophagocytic lymphohistiocytosis after bone marrow transplantation from related and unrelated donors: a single-centre experience of 12 patients.
Haemophagocytic lymphohistiocytosis (HLH) is an autosomal recessive disease with histiocytic and lymphocytic infiltrations in multiple organs. Cure seems possible only by allogeneic bone marrow transplantation (BMT), but matched sibling donors (MSD) are restricted and high mortality rates are associated with BMT from unrelated donors (URD). We report on 12 consecutive HLH patients with an improved outcome following URD transplants. Eight patients received BMT from URD, four from MSD. Five patients had signs of active HLH at the time of BMT. The conditioning regimen consisted of 20 mg/kg busulphan, 60 mg/kg VP-16 and 120 mg/kg cyclophosphamide and, in case of URD, 90 mg/kg antithymocyte globulin. The doses of busulphan and VP-16 were reduced during the programme to 16 mg/kg and 30 mg/kg, respectively. Using a fivefold graft-versus-host disease (GVHD) prophylaxis, GVHD was absent or mild in 10, and moderate or severe in two patients undergoing unrelated transplants. One patient with URD experienced graft failure and was retransplanted on day 37. Major toxicities were hepatic veno-occlusive disease in five, capillary leak syndrome in two, pneumonia in three, sepsis in one, severe mucositis in one and seizures in two patients. All patients are alive without HLH after a median follow-up of 24.5 months. One patient has chronic GVHD, another patient has severe retardation. Three patients show slight to moderate development delay. These results indicate that in HLH, BMT from matched unrelated donors should be performed. Incomplete resolution of disease activity need not impede a successful outcome.


Psychiatric morbidity in children with KCNJ11 neonatal diabetes.
Mutations in the KCNJ11 gene, which encodes the Kir6.2 subunit of the pancreatic KATP channel, cause neonatal diabetes. KCNJ11 is also expressed in the brain, and ~ 20% of those affected have neurological features, which may include features suggestive of psychiatric disorder. No previous studies have systematically characterized the psychiatric morbidity in people with KCNJ11 neonatal diabetes. We aimed to characterize the types of psychiatric disorders present in children with KCNJ11 mutations, and explore their impact on families. The parents and teachers of 10 children with neonatal diabetes due to KCNJ11 mutations completed the Strengths and Difficulties Questionnaire and the Development and Wellbeing Assessment. Strengths and Difficulties Questionnaire scores were compared with normative data. Diagnoses from the Development and Wellbeing Assessment were compared with known clinical diagnoses. Strengths and Difficulties Questionnaire scores indicated high levels of psychopathology and impact. Psychiatric disorder(s) were present in all six children with the V59M or R201C mutation, and the presence of more than one psychiatric disorder was common. Only two children had received a formal clinical diagnosis, with a further one awaiting assessment, and the coexistence of more than one psychiatric disorder had been missed. Neurodevelopmental (attention deficit hyperactivity disorder and autism) and anxiety disorders predominated. Systematic assessment using standardized validated questionnaires reveals a range of psychiatric morbidity in children with KCNJ11 neonatal diabetes. This is under-recognized clinically and has a significant impact on affected children and their families. An integrated collaborative approach to clinical care is needed to manage the complex needs of people with KCNJ11 neonatal diabetes.


Skeletal muscle protein synthesis and degradation in vitro: effects of temperature.
We compared the structure, function, protein synthesis, and degradation of 70- to 95-mg rat soleus muscles during 120 min of incubation at 20 and 37 degrees C. At 37 degrees C, muscles were characterized by a damaged central core region and a decline of isometric tension development during incubation. Protein synthesis in the core region at 37 degrees C was depressed relative to the peripheral region. At 20 degrees C, developed tension remained constant during incubation, and synthesis rates in the core region were not different from the peripheral region. Compared with fresh muscle, ATP concentration after incubation was not affected by temperature. After equilibration of phenylalanine specific activity between extracellular and intracellular spaces (60 min at 20 degrees C; 30 min at 37 degrees C), rates of protein synthesis at 20 [0.048 nmol tyrosine (Tyr) X mg wet mass-1 X 2 h-1] and 37 degrees C (0.160 nmol Tyr X mg wet mass-1 X 2 h-1) were linear up to 180 and 120 min, respectively. Rates of protein degradation at 20 (0.076 nmol Tyr X mg wet mass-1 X 2 h-1) and 37 degrees C (0.248 nmol Tyr X mg wet mass-1 X 2 h-1) measured after 60 min were linear up to 180 and 120 min, respectively. Incubation at 20 degrees C offers an approach to study 70- to 95-mg muscles in vitro without compromising structure and function.


Origin, growth, and rupture of saccular aneurysms: a review.
Pathological and hemodynamic concepts regarding the origin, growth, and rupture of intracranial saccular aneurysms are reviewed. Aneurysms form as a result of an interplay between hemodynamic factors, such as axial stream impingement and the water hammer effect, and structural weaknesses at apices of arterial bifurcations, such as congenital and acquired medial defects, funnel-shaped dilatations, and areas of thinning. Hypertension and time aid the formation of aneurysms. Unknown factors in women and in some families also play a role. Enlargement of aneurysms results from an interplay between mechanical factors, such as self-excitation and resonance, that produce structural fatigue and pathological processes of repair of the aneurysmal wall. Rupture of aneurysms is caused by the same hemodynamic factors that effect growth and is also influenced by extramural pressure. Pathologically, a major rupture may be preceded by fibrinous and leukocytic infiltration of the wall, bleb formation, and a minor hemorrhage. Such minor leaks can be followed by healing and growth. Aneurysms that escape major hemorrhage or heal successfully after a hemorrhage can grow to giant proportions, but remain susceptible to rupture despite their size, unless they become completely thrombosed. Intramural thrombosis may be stimulated by minor leaks and is dependent upon the physical characteristics of aneurysms. Experimental, angiographic, and clinical studies that pertain to the origin, growth, and rupture of aneurysms are also reviewed.


Pulmonary rehabilitation in Canada: A report from the Canadian Thoracic Society COPD Clinical Assembly.
Pulmonary rehabilitation (PR) is a recommended intervention in the management of individuals with chronic lung disease. It is important to study the characteristics and capacity of programs in Canada to confirm best practices and identify future areas of program improvement and research. To identify all Canadian PR programs, regardless of setting, and to comprehensively describe all aspects of PR program delivery. The present article reports the results of the survey related to type of program, capacity and program characteristics. All hospitals in Canada were contacted to identify PR programs. A representative from each program completed a 175-item online survey encompassing 16 domains, 10 of which are reported in the present article. A total of 155 facilities in Canada offered PR, of which 129 returned surveys (83% response rate). PR programs were located in all provinces, but none in the three territories. Most (60%) programs were located in hospital settings, 24% were in public health units and 8% in recreation centres. The national capacity of programs was estimated to be 10,280 patients per year, resulting in 0.4% of all Canadians with chronic obstructive pulmonary disease (COPD) and 0.8% of Canadians with moderate to severe COPD having access to PR. COPD, interstitial lung disease, and asthma were the most common diagnoses of patients. The majority of programs had at least four health care professionals involved; 9% had only one health care professional involved. The present comprehensive survey of PR in Canada reports an increase in the number of programs and the total number of patients enrolled since the previous survey in 2005. However, PR capacity has not kept pace with demand, with only 0.4% of Canadians with COPD having access.


De-escalation of surgery for early oral cancer--is it oncologically safe?
This study is a review of practice for patients with T1 or T2 squamous cell carcinoma (SCC) of the anterior tongue and floor of the mouth who presented to the regional maxillofacial unit in Liverpool between 1992 and 2007. We examined trends in management and analysed their effects on resection margins, recurrence, and survival. The Liverpool head and neck oncology database was used to identify patients, and to retrieve their clinical, surgical, and pathological data. When data were missing the case notes and pathology records were reviewed. Follow up was taken to January 2011. A total of 382 patients were included. Despite more conservative treatment with closer resection margins (27% in 1992-1995 and 60% in 2004-2007), fewer free flaps (79% in 1992-1995 and 38% in 2004-2007), and less adjuvant radiotherapy (37% in 1992-1995 and 22% in 2004-2007), there has been no significant increase in local recurrence (14% in 1992-1996 and 8% in 2004-2007), and overall survival has not been adversely affected. This is most striking when T1 tumours are considered in isolation with a consistent trend towards fewer clear margins (95% in 1992-1995 and 28% in 2004-2007) and fewer free flaps (53% in 1992-1995 and 11% in 2004-2007). The case mix was similar over the study period. These data support a more conservative approach to the management of early oral cancer.