Transcriptome analysis of coding and long non-coding RNAs highlights the regulatory network of cascade initiation of permanent molars in miniature pigs.
In diphyodont mammals, the additional molars (permanent molars) bud off from the posterior-free end of the primary dental lamina compared with successional teeth (replacement teeth) budding off from the secondary dental lamina. The diphyodont miniature pig has proved to be a valuable model for studying human molar morphogenesis. The additional molars show a sequential initiation pattern related to the specific tooth development stage of additional molars in miniature pigs during the morphogenesis of additional molars. However, the molecular mechanisms of the regulatory network of mRNAs and long non-coding RNAs during sequential formation of additional molars remain poorly characterized in diphyodont mammals. Here, we performed RNA-seq and microarray on miniature pigs at three key molar developmental stages to examine their differential gene expression profiles and potential regulatory networks during additional molar morphogenesis. We have profiled the differential transcript expression and functional networks during morphogenesis of additional molars in miniature pigs. We also have identified the coding and long non-coding transcripts using Coding-Non-Coding Index (CNCI) and annotated transcripts through mapping to the porcine, Wuzhishan miniature pig, mice, cow and human genomes. Many new unannotated genes plus 450 putative long intergenic non-coding RNAs (lincRNAs) were identified. Detailed regulatory network analyses reveal that WNT and TGF-β pathways are critical in regulating sequential morphogenesis of additional molars. This is the first study to comprehensively analyze the spatiotemporal dynamics of coding and long non-coding transcripts during morphogenesis of additional molars in diphyodont mammals. The miniature pig serves as a large model animal to elucidate the relationship between morphogenesis and transcript level during the cascade initiation of additional molars. Our data provide fundamental knowledge and a basis for understanding the molecular mechanisms governing cascade initiation of additional molars, but also provide an important resource for developmental biology research.


Using optoacoustic imaging for measuring the temperature dependence of Grüneisen parameter in optically absorbing solutions.
Grüneisen parameter is a key temperature-dependent physical characteristic responsible for thermoelastic efficiency of materials. We propose a new methodology for accurate measurements of temperature dependence of Grüneisen parameter in optically absorbing solutions. We use two-dimensional optoacoustic (OA) imaging to improve accuracy of measurements. Our approach eliminates contribution of local optical fluence and absorbance. To validate the proposed methodology, we studied temperature dependence of aqueous cupric sulfate solutions in the range from 22 to 4 °C. Our results for the most diluted salt perfectly matched known temperature dependence for the Grüneisen parameter of water. We also found that Grüneisen-temperature relationship for cupric sulfate exhibits linear trend with respect to the concentration. In addition to accurate measurements of Grüneisen changes with temperature, the developed technique provides a basis for future high precision OA temperature monitoring in live tissues.


Evidence for a role of p38 kinase in hypoxia-inducible factor 1-independent induction of vascular endothelial growth factor expression by sodium arsenite.
Recently we have demonstrated that sodium arsenite induces the expression of hypoxia-inducible factor 1alpha (HIF-1alpha) protein and vascular endothelial growth factor (VEGF) in OVCAR-3 human ovarian cancer cells. We now show that arsenic trioxide, an experimental anticancer drug, exerts the same effects. The involvement of phosphatidylinositol 3-kinase and mitogen-activated protein kinase (MAPK) pathways in the effects of sodium arsenite was investigated. By using kinase inhibitors in OVCAR-3 cells, both effects of sodium arsenite were found to be independent of phosphatidylinositol 3-kinase and p44/p42 MAPKS but were attenuated by inhibition of p38 MAPK. A role for p38 in the regulation of HIF-1alpha and VEGF expression was supported further by analysis of activation kinetics. Experiments in mouse fibroblast cell lines, lacking expression of c-Jun N-terminal kinases 1 and 2, suggested that these kinases are not required for induction of HIF-1alpha protein and VEGF mRNA. Unexpectedly, sodium arsenite did not activate a HIF-1-dependent reporter gene in OVCAR-3 cells, indicating that functional HIF-1 was not induced. In agreement with this hypothesis, up-regulation of VEGF mRNA was not reduced in HIF-1alpha(-/-) mouse fibroblast cell lines. Altogether, these data suggest that not HIF-1, but rather p38, mediates induction of VEGF mRNA expression by sodium arsenite.


Solitaire Stent in the Treatment of Acute Ischemic Stroke with Large Cerebral Artery Occlusion.
To investigate the effect of mechanical thrombectomy with solitaire stent in the treatment of acute ischemic stroke with large cerebral artery occlusion. Fifteen acute ischemic stroke patients with a proximal intracranial occlusion in the anterior circulation were included within 6 hours after symptom onset (unknown time of onset allowed in wake upstroke). Patients with a large infarct core or poor collateral circulation on computed tomography (CT) and CT angiography were excluded. All patients were measured by the National Institutes of Health Stroke Scale (NIHSS) before and 24 hours after the procedure. The primary outcomes were reperfusion at 24 hours and a Thrombolysis in Cerebral Infarction (TICI) score of 2b or 3 indicates successful reperfusion. Secondary outcomes included the functional score on the Modified Rankin Scale(MRS) and NIHSS score at 90 days. Good Functional Outcome (GFO), is defined as mRS0-2 at 90 days. The preoperative TICI grading of these 15 patients were all level 0.14 patients were level 2b to level 3 after the thrombectomy, 1 ipsilateral cervical carotid occlusion patient failed recanalization.14 patients with reperfusion at 24 hours showed good early neurologic improvement. The MRS score of all the 14 patients were<2 point at 90 days. There were no obvious adverse reactions and complications in these patients after mechanical thrombectomy. The application of mechanical thrombectomy with solitaire stent for the treatment of acute ischemic stroke with large cerebral artery occlusion is safe and time-efficient, which could improve the recanalization rate, decrease or even eliminate the application of thrombolytic drugs and reduce the rate of intracranial hemorrhage. Awake stroke patient can also benefit from thrombectomy.


Bronchial clearance of DTPA is increased in acute asthma but not in chronic asthma.
To investigate bronchial permeability in asthma, we measured the bronchial clearance of 113mIn-DTPA in seven asthmatics during and after an acute attack of asthma, seven asthmatics with chronic airflow limitation, and seven asthmatics without airflow limitation but with bronchial hyperresponsiveness to methacholine. We compared these results with those from seven normal subjects, seven patients with chronic bronchitis and bronchial infection, and seven patients with emphysema. An aerosol of 113mIn-DTPA was produced with a spinning disc to ensure a predominantly bronchial deposition of inhaled particles (6.3 microns MMAD). Radioactivity over the chest was recorded with a gamma-camera for 10 min after the subject inhaled the aerosol. Central regions of interest were selected, and the logarithm of the radioactivity was plotted against time; bronchial clearance of 113mIn-DTPA was calculated as the negative slope of the regression line. Clearance was substantially higher in asthmatics during their acute attacks than in all other groups (p less than 0.0001), and it decreased toward normal levels after recovery from the acute episode. The bronchial clearance of 113mIn-DTPA in all other groups did not differ from normal. We conclude that the bronchial clearance of 113mIn-DTPA is increased in asthmatics during attacks of asthma but in the stable state is not related either to bronchial hyperresponsiveness or to airflow limitation. Our findings are best explained by an increase in permeability of the bronchial mucosa of asthmatics during acute attacks.


Influence of fraction size on the development of late radiation enteropathy. An experimental study in the rat.
The use of large fraction sizes in radiotherapy may be associated with an increased risk of complications from late responding normal tissues. However, in the intestine, chronic injury may develop either as primary late effect or secondary to disruption of mucosal integrity as so-called consequential injury. Mucosal damage is relatively less sensitive to changes in fraction size than late reacting, slowly proliferating cells. The relationship between fraction size and chronic radiation enteropathy in a given situation may thus depend on which of the two mechanisms that predominates. Most previous studies of the influence of fraction size on radiation injury are confounded by differences in treatment time. The present study was therefore designed to assess subacute and chronic radiation enteropathy after three different fractionation regimens where fraction size was the only experimental variable. A total of 96 male Sprague-Dawley rats were orchiectomized and a functionally intact loop of small intestine was transposed into the left scrotum. These 'scrotal hernias' containing intestine were subsequently exposed to 50.4 Gy localized fractionated irradiation over 18 days with either 2.8 Gy every 24 h, 4.2 Gy every 36 h, or 5.6 Gy every 48 h. Control animals were sham irradiated. The animals were observed for development of intestinal complications (intestinal obstruction or enterocutaneous fistula formation) up to 6 months after irradiation. Histologic damage was assessed in groups of animals at 2 weeks (subacute injury) and 26 weeks (chronic injury), using a previously validated radiation injury score (RIS). RIS increased significantly with increasing fraction size at both observation times. However, the increase was more pronounced at 26 weeks than at 2 weeks. Increased chronic injury was characterized by increased incidence and severity of mucosal ulceration, serosal thickening, vascular sclerosis and intestinal wall fibrosis. We conclude that increasing fraction size increases both subacute and, even more markedly, chronic injury in the intestine. With the fractionation regimens used here, the chronic radiation enteropathy develops as a combined consequential and primary late effect, but the primary mechanism predominates.


16S rRNA Gene Amplicon Sequencing Reveals Significant Changes in Microbial Compositions during Cyanobacteria-Laden Drinking Water Sludge Storage.
This is the first study to systematically investigate the microbial community structure in cyanobacteria-laden drinking water sludge generated by different types of coagulants (including AlCl3, FeCl3, and polymeric aluminum ferric chloride (PAFC)) using Illumina 16S rRNA gene MiSeq sequencing. Results show that Cyanobacteria, Proteobacteria, Firmicutes, Bacteroidetes, Verrucomicrobia, and Planctomycetes were the most dominant phyla in sludge, and because of the toxicity of high Al and Fe level in AlCl3 and FeCl3 sludges, respectively, the PAFC sludge exhibited greater microbial richness than that in AlCl3 and FeCl3 sludges. Due to lack of light and oxygen in sludge, relative abundance of the dominant genera Microcystis, Rhodobacter, Phenylobacterium, and Hydrogenophaga clearly decreased, especially after 4 days storage, and the amounts of extracellular microcystin and organic matter rose. As a result, the relative abundance of microcystin and organic degradation bacteria increased significantly, including pathogens such as Bacillus cereus, in particular after 4 days storage. Hence, sludge should be disposed of within 4 days to prevent massive growth of pathogens. In addition, because the increase of extracellular microcystins, organic matter, and pathogens in AlCl3 sludge was higher than that in FeCl3 and PAFC sludges, FeCl3 and PAFC may be ideal coagulants in drinking water treatment plants.


Effect of Temperature, Oxygen, and Gibberellic Acid on the Development of Photosensitivity in Oldenlandia corymbosa L. Seeds during Their Incubation in Darkness.
Two successive phases can be distinguished in the development of the responsiveness to light in Oldenlandia corymbosa L. seeds during their incubation in darkness. During phase I, the responsiveness to light increases with time if there is sufficient O(2), and the higher the temperature, the faster the increase. This phase is stimulated by gibberellic acid. During the following phase (II), seeds remain responsive to light at 10 or 20 degrees C, but lose their responsiveness at higher temperature (>/=30 degrees C). This second phase depends on O(2): loss of responsiveness is accelerated at lower O(2) concentration. Phase II is only slightly affected by gibberellic acid. The results are discussed in terms of variation of phytochrome and of a reaction along the transduction chain initiated by phototransformation of this pigment, which is finally expressed in germination.


The Effects of Geometry Size and Initial Microstructure on Deformation Behavior of Electrically-Assisted Micro-Compression in Ti-6Al-4V Alloy.
In this study, electrically-assisted micro-compression (EAMC) tests were conducted for cylindrical specimens of Ti-6Al-4V alloy with four geometric sizes and three initial microstructures. The result showed that the specimen temperature nonlinearly increased with the square of current density. The quasi-static heat equilibrium equation was established to quantify the effects of the scale factor on the Joule heat temperature. Moreover, it was demonstrated that the Joule temperature scale effect had a greater effect on the flow stress than the sample size effect for specimens of different dimensions. It was noted that the 0.5 mm diameter sample displayed abnormal deformation behavior, which was related to surface oxidation leading a brittle surface layer. By comparison of the microstructures, it was found that the α→β phase transformation occured below the β transus temperature, which was attributed to the local Joule heat effect and the scattering of drift electrons during EAMC. Furthermore, the flow curves showed a strong dependence of the strength and ductility on the initial microstructure. The widmannstatten microstructure exhibited higher strength, smaller hardening rate and more easy flow localization compared with basket-weave microstructures, which was attributed to the low β phase content and narrow interlamellar spacing of α lamellae grains in the widmannstatten microstructure.


Transcriptomic changes during caste development through social interactions in the termite Zootermopsis nevadensis.
One of the most striking examples of phenotypic plasticity is the different phenotypes (i.e., castes) within a same nest of social insects. Castes usually derive from a single genotype initially by receiving social cues among individuals during development. Specific gene expression changes may be involved in caste differentiation, and thus, the regulatory mechanism of these changes should be clarified in order to understand social maintenance and evolution. The damp-wood termite Zootermopsis nevadensis is one of the most important model termite species, due to not only the availability of genomic, transcriptomic, and epigenomic information but also evidence that soldier- and worker-destined individuals can be identified in natural conditions. Given that the nutritional intakes via social interactions are crucial for caste differentiation in this species, there is a possibility that transcriptomic changes are influenced by the nutritional difference among these individuals. Here, whole body RNA-seq analysis of 3rd-instar larvae with biological replications and Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were conducted. We found the drastic expression differences during caste developments between soldier- and worker-destined individuals. The results indicated that there are several key signaling pathways responsible for caste formations, which are involved in developments and social interactions. Particularly, the nutritional sensitive signaling was upregulated in soldier-destined individuals, while some metabolic pathways were identified in worker-destined individuals. These bioinformatic data obtained should be utilized to examine the molecular mechanisms of caste determination in social insects.


Prognostic impact of transcatheter mitral valve repair in patients with exercise-induced secondary mitral regurgitation.
Although exercise-induced secondary mitral regurgitation (MR) is known to have a poor prognosis, the therapeutic strategy towards this condition remains to be investigated. In the present study, we aimed to investigate the prognostic impact of transcatheter mitral valve repair (TMVr) using the MitraClip in patients with exercise-induced secondary MR. Of the 200 consecutive patients with secondary MR who underwent exercise stress echocardiography, 46 (23%) that presented with exercise-induced secondary MR [i.e. increase in effective regurgitant orifice area (EROA) of ≥ 0.13 cm2] were enrolled in the present investigation. The composite endpoints of all-cause mortality and hospitalization for heart failure were evaluated. Of the 46 patients included in the current cohort, 19 (41%) underwent TMVr and 27 (59%) were medically managed (control group). Although the TMVr group tended to present with a greater EROA at rest (0.26 ± 0.10 vs. 0.20 ± 0.08 cm2, P = 0.047), there were no differences in the EROA changes during exercise between the two groups (0.18 ± 0.10 vs. 0.18 ± 0.04 cm2, P = 0.940). While the TMVr group reported a higher event-free survival rate after the 13-month follow-up period (log-rank P = 0.017), the Cox proportional-hazard analysis suggested the TMVr to be associated with clinical outcomes (hazard ratio: 0.419, P = 0.044). As opposed to the medical management, TMVr treatment was associated with a lower risk of composite endpoints in patients with exercise-induced secondary MR. Exercise stress echocardiography is considered to have played an important role in decision-making for secondary MR.


A review on fascioliasis in Egypt.
Fascioliasis, caused by Fasciola species, is a disease of herbivorous animals. It has a worldwide distribution in a large variety of grass-grazing animals as sheep, goats, cattle, buffaloes, horses and rabbits. In Egypt, donkeys and camels as well, are hosts for F. gigantica. Fascioliasis may occasionally affect man. Human infection causes serious hepatic pathological sequences that add to the already known threats to the liver of the Egyptian population. Two clinical stages are recognized in human fascioliasis. An acute stage coincides with the larval migration and worm maturation in the hepatic tissue, and a chronic stage coincides with the persistence of Fasciola worms in the bile ducts. Human infection with fascioliasis was very sporadic until the last three decades where clinical cases and outbreaks were reported. The estimated the number of people currently having fascioliasis to be 360,000 in Bolivia, 20,000 in Ecuador, 830,000 in Egypt, 10,000 in Islamic Republic of Iran, 742,000 in Peru, and 37,000 in Yemen. The total estimated number of people infected is 2.4 million in 61 countries and that the number at risk is more than 180 million throughout the world. Human fascioliasis has to be differentially diagnosed from some diseases as acute hepatitis, infection with other liver flukes as schistosomiasis, visceral toxocariasis, biliary tract diseases and hepatic amoebiasis. The parasitological diagnosis is based on identification of eggs in stool, duodenal contents or bile, also by the recovery of adult worm during surgical exploration, after treatment or at autopsy. However, the eggs may be present in very small number at irregular intervals, hence difficult to be found. Besides, the eggs may be transiently present in stool after ingestion of raw or undercooked liver from infected animals. The direct methods of diagnosing the egg are usually unsatisfactory. The symptoms may be present for several weeks before eggs are recovered in stool. Thus, the serologic tests are the alternative method of confirming early and extrabiliary human fascioliasis. However, cross-reactions with other helminthic antigen may confuse the interpretation of the results.


Peptide Coassembly to Enhance Piezoelectricity for Energy Harvesting.
The discovery of piezoelectricity in self-assembled peptide nanostructures opens an avenue to a new regime of piezoelectric materials and enables the fundamental investigation of electromechanical coupling in biomaterials. However, strategies for fabricating peptides with desired properties are still lacking. We find that a peptide-based coassembly process effectively controls the properties of peptide nanomaterials and demonstrates their application potential in nanogenerators. The composing peptides and their concentration influence the morphology, molecular property, and physical property of coassembled crystals. Compared with self-assembled diphenylalanine peptides, the coassembled peptides of diphenylalanine and phenylalanine-tryptophan show a 38% increase in piezoelectric coefficient, and the resulting harvesting device shows nearly a 3-fold increase in open-circuit voltage outputs.


Accuracy of High- and Low-Resolution Cone-Beam Computed Tomographic Scans in the Detection of Impacted Tooth-Induced External Root Resorption: An Ex-Vivo Study.
Cone-beam computed tomography (CBCT) is used in diagnostic situations, as well as tooth impaction and its complications. A possible sequela of tooth impaction is resorption of adjacent teeth, complicating the treatment plans. This study aimed to determine the diagnostic accuracy of high- and low-resolution CBCT scans in the detection of external root resorptions (ERRs), caused by an adjacent impacted tooth in the cementoenamel junction (CEJ), mid-root, and apical areas. Forty-five intact single-rooted teeth were divided into three groups of 15. Each group was dedicated to each zone of the root. Slight, moderate, and severe ERRs were formed, and CBCT scans were taken before and after the formation of ERRs. The diagnostic accuracy was assessed, and the Proportion test was used to compare the results. The statistical analyses of high- and low-resolution images showed a significant difference (P<0.05), which implies the higher accuracy of high-resolution images. The highest diagnostic accuracy among different zones was related to the mid-root, and the lowest was related to the apical zone. In terms of the size of ERRs, the diagnostic accuracy was the lowest for slight ERRs. The most reliable and accurate diagnostic mode was found in high-resolution images, in the mid-root zone, and with severe ERRs. The lowest diagnostic accuracy was found in low-resolution images, in the apical zone, and with slight ERRs.


The interspersed DNA repeat conserved in water birds' genomes.
A putative old and ubiquitous interspersed DNA repeat family was identified from TaqI restriction, M13 cloning, and sequencing of the genomic DNA of a Mallard (Anas platyrhynchos), a Muscovy Duck (Cairina moschata), a Toulouse Goose (Anser anser), and a Black Swan (Cygnus atratus). A 425-bp consensus core sequence was obtained for the interspersed family. The 425-bp unit was about 2% of the avian genome and was found to be conserved in at least four genera of the order Anseriforme: Anas, Anser, Cairina, and Cygnus.


Quality of life after stroke in old age: comparison of persons living in nursing home and those living in their own home.
To determine the differences in subjective quality of life between elderly people living in a nursing home and those living in their own homes after brain stroke, and to determine the contribution of demographic variables and different quality of life domains to the explanation of self-assessed quality of life. The study included 60 elderly men and women, 30 living in their own homes (median age, 81; range, 72-90) and 30 living in a nursing home (median age, 81; range, 72-86). Both groups received care (stationary or ambulatory) from the same nursing home. World Health Organization Quality of Life Questionnaire-short version, self-assessed quality of life questionnaire, and demographic questionnaire were used to collect data on subjective quality of life. The participants completed self-report questionnaires individually. Quality of life scores were significantly higher in the elderly living in a nursing home than in the elderly living in their own home (mean +/- standard deviation, 78.7 +/- 12.8 vs 59.3 +/- 17.3 out of maximum 100, P < 0.001). Also, the elderly living in the nursing home scored significantly higher than those living in their own home on all 4 quality of life domains (maximum 100 for each domain): physical (28.5 +/- 3.3 vs 17.2 +/- 5.0), psychological (22.3 +/- 3.7 vs 16.3 +/- 5.0), social relationships (11.4 +/- 1.6 vs 8.3 +/- 1.7), and environment (32.8 +/- 4.6 vs 24.0 +/- 6.1) domain (P < 0.001 for all). All predictive variables together explained 51.9% of quality of life variance, with self-assessed health being the most significant predictor. Quality of life of the elderly in a nursing home was significantly higher than that of their peers living in their own home, which may be related to better care in specially organized settings.


Direct selection for mutations affecting specific splice sites in a hamster dihydrofolate reductase minigene.
A Chinese hamster cell line containing an extra exon 2 (50 bp) inserted into a single intron of a dihydrofolate reductase (dhfr) minigene was constructed. The extra exon 2 was efficiently spliced into the RNA, resulting in an mRNA that is incapable of coding for the DHFR enzyme. Mutations that decreased splicing of this extra exon 2 caused it to be skipped and so produced normal dhfr mRNA. In contrast to the parental cell line, the splicing mutants display a DHFR-positive growth phenotype. Splicing mutants were isolated from this cell line after treatment with four different mutagens (racemic benzo[c]phenanthrene diol epoxide, ethyl methanesulfonate, ethyl nitrosourea, and UV irradiation). By polymerase chain reaction amplification and direct DNA sequencing, we determined the base changes in 66 mutants. Each of the mutagens generated highly specific base changes. All mutations were single-base substitutions and comprised 24 different changes distributed over 16 positions. Most of the mutations were within the consensus sequences at the exon 2 splice donor, acceptor, and branch sites. The RNA splicing patterns in the mutants were analyzed by quantitative reverse transcription-polymerase chain reaction. The recruitment of cryptic sites was rarely seen; simple exon skipping was the predominant mutant phenotype. The wide variety of mutations that produced exon skipping suggests that this phenotype is the typical consequence of splice site damage and supports the exon definition model of splice site selection. A few mutations were located outside the consensus sequences, in the exon or between the branch point and the polypyrimidine tract, identifying additional positions that play a role in splice site definition. That most of these 66 mutations fell within consensus sequences in this near-saturation mutagenesis suggests that splicing signals beyond the consensus may consist of robust RNA structures.


Reduced Severity of Tissue Injury Within the Infarct May Partially Mediate the Benefit of Reperfusion in Ischemic Stroke.
Emerging data suggest tissue within the infarct lesion is not homogenously damaged following ischemic stroke but has a gradient of injury. Using blood-brain-barrier (BBB) disruption as a marker of tissue injury, we tested whether therapeutic reperfusion improves clinical outcome by reducing the severity of tissue injury within the infarct in patients with ischemic stroke. In a pooled analysis of patients treated for anterior circulation large vessel occlusion in the EXTEND-IA TNK (Tenecteplase Versus Alteplase Before Endovascular Therapy for Ischemic Stroke) and EXTEND-IA part-2 (Determining the Optimal Dose of Tenecteplase Before Endovascular Therapy for Ischaemic Stroke) trials, post-treatment BBB permeability at 24 hours was calculated based on the extent of T1-brightening by extravascular gadolinium on T2* perfusion-weighted imaging and measured within the diffusion-weighted-imaging lesion. First, to determine the clinical significance of BBB disruption as a marker of severity of tissue injury, we examined the association between post-treatment BBB permeability and functional outcome. Second, we performed an exploratory (reperfusion, BBB permeability, functional outcome) mediation analysis to estimate the proportion of the reperfusion-outcome relationship that is mediated by change in BBB permeability. In the 238 patients analyzed, an increased BBB permeability measured within the infarct at 24 hours was associated with a reduced likelihood of favorable outcome (90-day modified Rankin Scale score of ≤2) after adjusting for age, baseline National Institutes of Health Stroke Scale, premorbid modified Rankin Scale, infarct topography, laterality, thrombolytic agent, sex, parenchymal hematoma, and follow-up infarct volume (adjusted odds ratio, 0.86 [95% CI, 0.75-0.98]; P=0.023). Mediation analysis suggested reducing the severity of tissue injury (as estimated by BBB permeability) accounts for 18.2% of the association between reperfusion and favorable outcome, as indicated by a reduction in the regression coefficient of reperfusion after addition of BBB permeability as a covariate. In patients with ischemic stroke, reduced severity of tissue injury within the infarct, as determined by assessing the integrity of the BBB, is independently associated with improved functional outcome. In addition to reducing diffusion-weighted imaging-defined infarct volume, reperfusion may also improve clinical outcome by reducing tissue injury severity within the infarct.


Risk of new or recurrent cancer under immunosuppressive therapy in patients with IBD and previous cancer.
To explore the risk of new or recurrent cancer among patients with IBD and previous cancer, exposed or not to immunosuppressants. Among the 17 047 patients of the CESAME prospective observational cohort who were enrolled from May 2004 to June 2005, and followed-up until December 2007, we identified 405 patients with cancer diagnosed previous to study entry. We calculated the rates of incident cancer in patients with or without previous cancer, and we assessed by survival analysis and nested case-control study the impact of immunosuppressants on the risk of incident new or recurrent cancer in patients with previous cancer. The rate of incident cancer was 21.1/1000 patient-years (PY) and 6.1/1000 PY in patients with and without previous cancer, respectively. The multivariate-adjusted HR of incident cancer between patients with and without previous cancer was 1.9 (95% CI 1.2 to 3.0, p=0.003). Among patients with previous cancer, the rates of new and recurrent cancers were, respectively, 13.2/1000 PY and 6.0/1000 PY in the 312 patients who were not taking immunosuppressant at the time of study entry, and 23.1/1000 PY and 3.9/1000 PY in the 93 patients treated with immunosuppressants at study entry. There was no significant association between the exposure to immunosuppressants and the risk of new or recurrent cancer. Patients with IBD with a history of cancer are at increased risk of developing any (new or recurrent) cancer, with a predominant incidence of new cancers. Treatment with immunosuppressants has no overall major impact per se on this risk.


Contact pressure of the coracoacromial arch in shoulders with joint contracture: A cadaveric study.
There have been many reports describing that the capsular fibrosis of the shoulder joint is the main cause of frozen shoulder, whereas others reported the significance of subacromial impingement as an etiological factor. The purpose of this study was to investigate the contact pressure between the coracoacromial arch and the rotator cuff tendons to clarify the contact phenomenon in shoulders with joint contracture. Fourteen fresh-frozen cadaveric shoulders were used. Specimens were divided into two groups: normal group (8 shoulders, definition: more than 61° of flexion and abduction and more than 21° of external rotation) and joint contracture group (6 shoulders, definition: less than 60° of flexion and abduction and less than 20° of external rotation). Contact pressure and area beneath the coracoacromial arch were measured by a flexible force sensor during flexion, abduction, internal and external rotation in adduction and abduction, extension and horizontal extension motions. The peak contact pressure under the acromion was observed at 90° in flexion and abduction in the normal group, whereas that in the contracture group was observed at 30° in flexion (P = 0.037) and at 30° in abduction (P = 0.041). Contact pressure in the contracture group was significantly higher than that in the normal group at 20° and 30° of abduction (P = 0.043, P = 0.041, respectively). There were no significant differences of contact pressure during other motions. Although no significant differences of contact pressure beneath the coracoacromial ligament were observed, contact area significantly increased in extension and horizontal extension motion. The contact between the acromion and the rotator cuff was observed in lower angles of flexion and abduction in shoulders with contracture than in those without. When treating patients with shoulder contracture, we need to perform rehabilitation taking such an abnormal movement into consideration.


[At-home-therapeutic space. Development of a device intended for family therapy at home].
Inspired by his ethnographic experience with the Mapuche, a native community in the South of Chile and his reflection on family therapy and the literature pertaining to ethnopsychiatry, the author proposes the elaboration of an at-home-therapeutic space (HTS), a specific device for family therapy in the homes of patients. The author describes the various steps of his approach and his first-hand experience of the device.


Glycogen synthase kinase-3beta regulates etoposide-induced apoptosis via Bcl-2 mediated caspase-3 activation in C3H10T1/2 cells.
Glycogen synthase kinase-3beta (GSK3beta) controls the survival of osteoblasts during bone development through Wnt canonical signaling. GSK3beta is a key factor for osteoblastogenesis, but relatively less is known regarding its role in osteoblast apoptosis. Genotoxic stress induced by etoposide promoted apoptotic signaling by GSK3beta activation in C3H10T1/2 cells, a mouse mesenchymal cell line. Etoposide led to the time-dependent activation of GSK3beta and caspase-3, which resulted in PARP cleavage. LiCl (a specific inhibitor) and siRNA (gene knock-down) of GSK3beta prevented the effects of etoposide on apoptosis. Staurosporine also induced apoptosis in C3H10T1/2 cells, but LiCl could not rescue. Bcl-2 was decreased in the cells by exposure to etoposide. LiCl completely recovered Bcl-2 expression as shown by both the mRNA and the protein expression levels. In conclusion, etoposide-induced apoptosis in C3H10T1/2 cells is mediated by GSK3beta, which leads to caspase-3 activation via decrease in Bcl-2 expression.


Effects of 4-aminopyridine in experimental CNS demyelination.
We studied the effect of systemically administered 4-aminopyridine in a model of CNS demyelination. In five rats with demyelination, slowed conduction velocity through the lesion was partially reversed at dose levels of 5.6 to 7.2 mg/kg. All rats developed convulsion at this dosage, and impaired conduction of high-frequency impulses was unchanged. These findings suggest certain limitations of 4-aminopyridine as a therapeutic agent.


Parental and practitioner perspectives on raising an adolescent with a disability: a focus group study.
To examine the challenges faced by parents of teenagers with a disability to determine the need for a tailored parenting program for this population. Focus groups were conducted with six parents and nine practitioners with experience supporting parents of teenagers with a disability. An inductive thematic analysis revealed close agreement between parents and practitioners and eight understandable and relevant themes. The results indicated that parents have difficulty planning and facilitating transitions for their adolescent children, managing behavioural problems due to these problems being unique to this developmental period and because some of the parenting strategies that were useful when their children were younger were no longer applicable. High levels of stress and feelings of grief were also described. These results suggest that parents of adolescents with a developmental disability could benefit from a parenting program and that an existing evidence-based parenting program should be modified. Implications for program design for this population are discussed. Parents have difficulty supporting their teenagers' behaviour. Some parenting strategies that were useful for young children are no longer applicable. Parents may benefit from a parenting program tailored for teenagers.


Ospapst1, a useful mutant for identifying seed purity and authenticity in hybrid rice.
The stability and completeness of male sterility is still a challenge in some male sterile rice lines, especially those of photoperiod/thermo-sensitive genic male sterility (P/TGMS). Leaf color marker is a widely practiced approach to reduce the impact of self-pollinated seeds of male sterile lines. The papst1 is a leaf color mutant. The newly emerged leaves of papst1 are chlorosis and have an impaired photosynthesis. But the other agronomic traits, such as germination rate, duration of maturation and seed weight, are not changed. The papst1/PAPST1 F₁ showed the wild-type leaf phenotype. The papst1/PAPST1 F₂ progenies displayed an approximately 3:1 segregation ratio of WT phenotype:mutant phenotype (72: 28, χ(2) = 0.48, p > 0.05), suggesting that papst1 mutant phenotype is caused by a single repressive gene. Map-based cloning and sequencing analysis revealed that a point mutation was occurred in Os01 g16040 (OsPAPST1). Given these results, the Ospapst1 mutant is a useful mutant for identifying seed purity and authenticity in hybrid rice.


Four new C19-diterpenoid alkaloids from the roots of Aconitum ouvrardianum.
Four new C19-diterpenoid alkaloids, 3-dehydroxyl-lipoindaconitine (1), 8-dehydroxyl-bikhaconine (2), 19R-acetonyl-talatisamine (3), and 16-hydroxyl-vilmorisine (4), were isolated from the roots of Aconitum ouvrardianum. Their structures were elucidated by spectral analyses, including ESI-MS, HR-ESI-MS, IR, UV, 1D and 2D NMR.


Somatostatin and counterregulatory hormone responses to hypoglycaemia in diabetics with and without autonomic neuropathy.
Blood glucose, somatostatin and counterregulatory hormone responses to an i.v. bolus of insulin were studied in insulin-dependent diabetics with different degrees of autonomic neuropathy, after 24 hours of optimised control with an artificial pancreas. There was no plasma catecholamine response in patients with a sympathetic autonomic neuropathy. A normal somatostatin response to hypoglycemia was absent in patients with autonomic neuropathy. Glucagon did not respond in diabetics, independently of the degree of neuropathy. In all diabetics, cortisol and GH were stimulated. Absence of warning symptoms was observed in patients with catecholamine deficiency. Despite different hormone behaviour, blood glucose fall and recovery were similar in all diabetic groups. It is concluded that the glucagon response to insulin hypoglycaemia is reduced in all type 1 longstanding diabetics, whereas catecholamine and somatostatin responses are only abolished in those with autonomic neuropathy. Patients with sympathetic neuropathy would be considered at increased risk severe hypoglycaemia.


Pain experienced by infants and toddlers at urine collection bag removal: A randomized, controlled, clinical trial.
In pre-continent children, collection bags are frequently used as a first-line option to obtain a urine specimen. This practice, acknowledged by several guidelines for the step of UTI screening, is driven by a perception of the technique as being more convenient and less painful. However, our own experience led us to consider bag removal as a painful experience. Our aim was to determine whether the use of an oleo-calcareous liniment to aid bag removal reduced the acute pain expressed by young children. This prospective, randomized, controlled, single blind study was carried out in two emergency pediatrics departments. Pre-continent children aged 0-36 months admitted with an indication for urine testing were eligible for the study. Urine for dipstick test screening was obtained using a collection bag. At micturition, the patients were randomized into bag removal with (intervention group) or without (control group) liniment. Bag removal was recorded on video in such a manner as to permit independent assessments of pain by two evaluators blinded to group allocation. Pain was assessed using the FLACC scale. 135 patients were analyzed: 70 in the intervention group and 65 in the control group. The median FLACC scores [interquartile range] for the intervention and control groups, respectively 4.0 [2.0-7.0] and 4.0 [3.0-7.0], did not differ significantly (p = 0.5). A FLACC score ≥4 was obtained for 56% of the patients and a score ≥7 for 28%. Removal of urine collection bags caused moderate to severe pain in half of the children included. The use of an oleo-calcareous liniment did not reduce this induced pain.


Granulomatosis with polyangiitis and associated pulmonary emphysema: Breathtaking vasculitis.
Pulmonary emphysema occasionally occurs in the absence of smoking or noxious exposures. Other than through a known association with alpha-1 antitryspin deficiency, to our knowledge, no reports implicate granulomatosis with polyangiitis (GPA) in causing airflow obstruction with small airway involvement and severe air trapping. To extend available experience, we report a 51-year-old male with biopsy-proven cytoplasmic-antineutrophilic cytoplasmic antibody proteinase 3 (ANCA PR3)-positive GPA who developed centrilobular emphysema and airflow obstruction during a phase of active vasculitis. He was a lifelong non-smoker and had a normal alpha-1 antitrypsin level and a PI*MM phenotype. Treatment with corticosteroids and cyclophosphamide caused clinical remission of his vasculitis which was associated with improvement in his respiratory symptoms. However, to date, structural changes of emphysema have persisted for over 9 years of follow-up. Clinicians should remain vigilant to the possibility of emphysema in patients with pulmonary vasculitis.


Comparison of Topical Chlorhexidine and Mupirocin for the Prevention of Exit-Site Infection in Incident Peritoneal Dialysis Patients.
♦ OBJECTIVE: Prevention of exit-site infection (ESI) is of paramount importance to peritoneal dialysis (PD) patients. The aim of this study was to evaluate the effectiveness of chlorhexidine in the prevention of ESI in incident PD patients compared with mupirocin. ♦ METHODS: This retrospective, pre-test/post-test observational study included all incident PD patients at Singapore General Hospital from 2012 to 2015. Patients received daily topical exit-site application of either mupirocin (2012 - 2013) or chlorhexidine (2014 - 2015) in addition to routine exit-site cleaning with 10% povidone-iodine. The primary outcome was ESI rate during the 2 time periods. Secondary outcomes were peritonitis rate, times to first ESI and peritonitis, hospitalization rate, and infection-related catheter removal. Event rates were analyzed using Poisson regression, and infection-free survival was estimated using Kaplan-Meier and Cox regression survival analyses. ♦ RESULTS: The study included 162 patients in the mupirocin period (follow-up 141.5 patient-years) and 175 patients in the chlorhexidine period (follow-up 136.9 patient-years). Compared with mupirocin-treated patients, chlorhexidine-treated patients experienced more frequent ESIs (0.22 vs 0.12 episodes/patient-year, p = 0.048), although this was no longer statistically significant following multivariable analysis (incidence rate ratio [IRR] 1.78, 95% confidence interval [CI] 0.98 - 3.26, p = 0.06). No significant differences were observed between the 2 groups with respect to time to first ESI (p = 0.10), peritonitis rate (p = 0.95), time to first peritonitis (p = 0.60), hospitalization rate (p = 0.21) or catheter removal rate (0.03 vs 0.04/patient-year, p = 0.56). ♦ CONCLUSIONS: Topical exit-site application of chlorhexidine cream was associated with a borderline significant, higher rate of ESI in incident PD patients compared with mupirocin cream.


The guinea-pig is not a rodent.
In 1991 Graur et al. raised the question of whether the guinea-pig, Cavia porcellus, is a rodent. They suggested that the guinea-pig and myomorph rodents diverged before the separation between myomorph rodents and a lineage leading to primates and artiodactyls. Several findings have since been reported, both for and against this phylogeny, thereby highlighting the issue of the validity of molecular analysis in mammalian phylogeny. Here we present findings based on the sequence of the complete mitochondrial genome of the guinea-pig, which strongly contradict rodent monophyly. The conclusions are based on cumulative evidence provided by orthologically inherited genes and the use of three different analytical methods, none of which joins the guinea-pig with myomorph rodents. In addition to the phylogenetic conclusions, we also draw attention to several factors that are important for the validity of phylogenetic analysis based on molecular data.


Intraabdominal abscesses: image-guided diagnosis and therapy.
Intraabdominal abscesses may complicate many illnesses including diverticulitis, pancreatitis, and appendicitis, or they may occur during the postoperative period. As new methods of imaging are developed that provide additional information on patients with these abscesses, earlier and more accurate diagnoses can be made, allowing for prompt intervention. With the advent of these new imaging methods, techniques for treatment of abscesses by percutaneous drainage have been developed. In light of these advances, we review the current strategies for the management of intraabdominal abscesses.


Evolutionary rescue via transgenerational plasticity: Evidence and implications for conservation.
When a population experiences severe stress from a changing environment, evolution by natural selection can prevent its extinction, a process dubbed "evolutionary rescue." However, evolution may be unable to track the sort of rapid environmental change being experienced by many modern-day populations. A potential solution is for organisms to respond to environmental change through phenotypic plasticity, which can buffer populations against change and thereby buy time for evolutionary rescue. In this review, we examine whether this process extends to situations in which the environmentally induced response is passed to offspring. As we describe, theoretical and empirical studies suggest that such "transgenerational plasticity" can increase population persistence. We discuss the implications of this process for conservation biology, outline potential limitations, and describe some applications. Generally, transgenerational plasticity may be effective at buying time for evolutionary rescue to occur.


[The habitat and environment-induced imbalance of trace elements in the populations in the population of urban and rural areas].
A comprehensive hygienic assessment was made of natural and anthropogenic risk factors and conditions for trace element deficiencies in the populations of urban and rural areas. All chemicals identified were evaluated in the environmental objects. The mean daily absorption dose for the population was estimated. The paper presents a comparative analysis of the concentrations of Cu, Fe, Zn, Pb, Mn, Ni, Cr, Co, Cd, Sr in the biological media (hair, blood, bone tissue, placenta) in the populations of an industrial town and rural areas and their levels in some abnormalities and also compares the spectral composition of trace elements in the hair and blood with that obtained by other authors.


An efficient tomato-detection method based on improved YOLOv4-tiny model in complex environment.
Automatic and accurate detection of fruit in greenhouse is challenging due to complicated environment conditions. Leaves or branches occlusion, illumination variation, overlap and cluster between fruits make the fruit detection accuracy to decrease. To address this issue, an accurate and robust fruit-detection algorithm was proposed for tomato detection based on an improved YOLOv4-tiny model. First, an improved backbone network was used to enhance feature extraction and reduce overall computational complexity. To obtain the improved backbone network, the BottleneckCSP modules of the original YOLOv4-tiny backbone were replaced by a Bottleneck module and a reduced version of BottleneckCSP module. Then, a tiny version of CSP-Spatial Pyramid Pooling (CSP-SPP) was attached to the new backbone network to improve the receptive field. Finally, a Content Aware Reassembly of Features (CARAFE) module was used in the neck instead of the traditional up-sampling operator to obtain a better feature map with high resolution. These modifications improved the original YOLOv4-tiny and helped the new model to be more efficient and accurate. The experimental results showed that the precision, recall, F 1 score, and the mean average precision (mAP) with Intersection over Union (IoU) of 0.5 to 0.95 were 96.3%, 95%, 95.6%, and 82.8% for the improved YOLOv4-tiny model, respectively. The detection time was 1.9 ms per image. The overall detection performance of the improved YOLOv4-tiny was better than that of state-of-the-art detection methods and met the requirements of tomato detection in real time.


Modulation of peripheral blood immune cells by early use of steroids and its association with clinical outcomes in patients with metastatic non-small cell lung cancer treated with immune checkpoint inhibitors.
Steroids are frequently used in patients with metastatic non-small cell lung cancer (mNSCLC), but they could be detrimental for patients treated with immune checkpoint inhibitors (ICIs). Here, we assessed the association between early use of steroids, clinical outcomes and peripheral immune blood cells modulation in patients with mNSCLC treated with ICIs. We reviewed patients with mNSCLC treated at our institution between April 2013 and December 2017. Early use of steroids was defined as the use of a daily prednisone-equivalent dose ≥10 mg for at least 1 day within 28 days after ICI initiation. Peripheral immune blood cell counts were retrieved at baseline and at 4 and 6 weeks after ICI initiation. Out of 151 patients included, 35 (23%) made early use of steroids that was associated with poor disease control (OR 0.32, p=0.006), progression-free survival (HR 1.80, p=0.003) and overall survival (HR 2.60, p<0.001). Early use of steroids significantly correlated with higher median absolute neutrophil count, neutrophil to lymphocyte ratio (NLR) and derived NLR, and lower median absolute and relative eosinophil count, both at 4 and 6 weeks after ICI initiation. In patients with mNSCLC treated with ICIs, early use of steroids was associated with worse clinical outcomes and remarkable modulation of peripheral blood immune cells, which could contribute to restraining the activation of antitumour immunity. If confirmed in prospective studies, these findings would highlight the importance of carefully evaluating and, whenever possible, avoiding steroids during early phases of ICI treatment.


Synthesis, Identification, and Structure Elucidation of Adducts Formed by Reactions of Hydroxycinnamic Acids with Glutathione or Cysteinylglycine.
Grape polyphenols, especially hydroxycinnamic acids such as caftaric and caffeic acid, are prone to enzymatic oxidation reactions during the winemaking process, forming o-quinones and leading to color darkening. Glutathione is capable of trapping these o-quinones and thus limiting juice browning. In this study, the addition of glutathione or cysteinylglycine onto caftaric or caffeic acid o-quinones formed by polyphenoloxidase-catalyzed reactions was investigated by UPLC-DAD-ESIMS and NMR data analyses. Complete identification of adducts has been achieved via NMR data. The results confirmed that the favored reaction is the substitution of the sulfanyl group of cysteine at C-2 of the aromatic ring. Several minor isomers, namely, the cis-isomer of the 2-S adduct and trans-isomers of the 5-S and 6-S adducts, and the 2,5-di-S-glutathionyl adducts were also identified and quantified by qNMR. With the exception of 2-(S-glutathionyl)- and 2,5-di(S-glutathionyl)-trans-caftaric acid, these products had never been formally identified. In particular, the 5-S and 6-S derivatives are reported here for the first time. The first formal identification of 2-S cis-derivatives is also provided. Moreover, NMR and UPLC-DAD-ESIMS analysis showed that signature UV and MS spectra can serve as markers of the conformation and substitution position in the aromatic ring for each of the isomers.


[Entecavir - options and obstacles of an effective treatment for chronic hepatitis B].
The level of HBV-DNA is a crucial determinant of the progression to liver cirrhosis or hepatocellular carcinoma. Thus the effective suppression of HBV-DNA below the limit of detection of high sensitive assays is a major aim of treatment. This aim will be achieved in almost all patients with current available direct antiviral drugs. However, the development of drug resistance remains a main challenge for the future. Entecavir has a good profile regarding both antiviral efficacy and resistance profile in the treatment of previously untreated patients. After five year on Entecavir more than 90 % of HBe-Ag positive patients had achieved HBV-DNA below 300 copies/ml and resistance developed in only about 1 %. However, patients in whom there had been previous resistance to lamivudine have lower treatment responses and higher resistance rates because only one or two additional mutations in the HBV-polymerase gene are required for the development of entecavir resistance in contrast to three mutations required in treatment naïve patients. In summary, Entecavir adds a new milestone in the treatment of chronic hepatitis B.


Psychosocial influences on help-seeking behaviour for cancer in low-income and lower middle-income countries: a mixed-methods systematic review.
Starting cancer treatment early can improve outcomes. Psychosocial factors influencing patients' medical help-seeking decisions may be particularly important in low and lower middle-income countries (LMIC) where cancer outcomes are poor. Comprehensive review evidence is needed to understand the psychosocial influences on medical help-seeking for cancer symptoms, attendance for diagnosis and starting cancer treatment. Mixed-methods systematic review registered on PROSPERO (CRD42018099057). Peer-reviewed databases were searched until April 2020 for studies assessing patient-related barriers and facilitators to medical help-seeking for cancer symptoms, diagnosis and treatment in adults (18+ years) living in LMICs. Quality of included studies was assessed using the Critical Appraisal Skills Programme tool. Data were synthesised using meta-analytic techniques, meta-ethnography or narrative synthesis as appropriate. Of 3963 studies identified, 64 were included. In quantitative studies, use of traditional, complementary and alternative medicine (TCAM) was associated with 3.60 higher odds of prolonged medical help-seeking (95% CI 2.06 to 5.14). Qualitative studies suggested that use of TCAM was a key barrier to medical help-seeking in LMICs, and was influenced by causal beliefs, cultural norms and a preference to avoid biomedical treatment. Women face particular barriers, such as needing family permission for help-seeking, and higher stigma for cancer treatment. Additional psychosocial barriers included: shame and stigma associated with cancer such as fear of social rejection (eg, divorce/disownment); limited knowledge of cancer and associated symptoms; and financial and access barriers associated with travel and appointments. Due to variable quality of studies, future evaluations would benefit from using validated measures and robust study designs. The use of TCAM and gender influences appear to be important barriers to help-seeking in LMIC. Cancer awareness campaigns developed with LMIC communities need to address cultural influences on medical help-seeking behaviour.


Antibacterial activities of two potential peptides extracted from Polistes wattii Cameron, 1900 (Vespidae: Polistinae) wasp venom collected at Eastern Province, Saudi Arabia.
Alternatives of conventional antibiotics have become an urgent need to control drug-resistant bacteria. Therefore, search for new antibacterial agents has become a trend in several microbiological and pharmaceutical scientific works. Insects, one of the most successful and evolved species on earth is known to be an effective natural source of several medically useful chemicals including antibacterial agents. There is considerable evidence of using wasp venom against medical ailments in several parts of the world. In this work venom from Polistes wattii Cameron, 1900 collected from Eastern Province, Saudi Arabia was evaluated for its antibacterial activities. Such activity was tested against four pathogenic bacteria: two-gram positive Staphylococcus aureus (ATCC 25923) and Streptococcus mutans (RCMB 017(1) ATCC 25175) and two gram-negative (Salmonella typhimurium NCTC 12023 ATCC 14028 and Enterobacter cloacae (RCMB 001(1) ATCC 23355). Also, chemical characterization of wasp venom was done using HPLC and two isolated peptides were sequenced. The result indicates the potent anti-microbial effect of the venom against the four tested bacteria. The most sensitive bacteria were Staphylococcus aureus (ATCC 25923) and Streptococcus mutans (RCMB 017(1) ATCC 25175). The sequence of the two purified peptides indicates that they belong to mastoparan. The study results may pave way to use this wasp venom in future antibiotics especially in controlling skin infection by Staphylococcus aureus.


[Incidence of otosclerosis in a subpopulation of Galicia, Spain].
We have made a prospective study of the patients who were analysed for the first time at the Outpatients Department in the Otolaryngology Section of the University Complex Hospital of Santiago de Compostela all along 1997. A whole of 1,050 patients have been included in this study. The most frequent diagnosis were neurosensorial hypoacusis (29.1%), chronic otitis media (27.8%) and in the third place, otosclerosis. This appeared in 83 cases, which represents a 8.7% of the fulfilled diagnosis. The incidence for our health area reached 19 new cases of otosclerosis per 100,000 inhabitants a year. When only the number of stapedectomies were included, it reached a 7.8%. The prevalence of the female population was clear, reaching a 77%. Diagnosed patients' age was from 30 to 50 years, although a considerable percentage (37%) were older. The incidence reached by our studies is higher to other values previously published. We think this responds to the fact that the health system covers all the population and to the importance a good audition is given nowadays.


Key issues in the design of pay for performance programs.
Pay for performance (P4P) is increasingly being used to stimulate healthcare providers to improve their performance. However, evidence on P4P effectiveness remains inconclusive. Flaws in program design may have contributed to this limited success. Based on a synthesis of relevant theoretical and empirical literature, this paper discusses key issues in P4P-program design. The analysis reveals that designing a fair and effective program is a complex undertaking. The following tentative conclusions are made: (1) performance is ideally defined broadly, provided that the set of measures remains comprehensible, (2) concerns that P4P encourages "selection" and "teaching to the test" should not be dismissed, (3) sophisticated risk adjustment is important, especially in outcome and resource use measures, (4) involving providers in program design is vital, (5) on balance, group incentives are preferred over individual incentives, (6) whether to use rewards or penalties is context-dependent, (7) payouts should be frequent and low-powered, (8) absolute targets are generally preferred over relative targets, (9) multiple targets are preferred over single targets, and (10) P4P should be a permanent component of provider compensation and is ideally "decoupled" form base payments. However, the design of P4P programs should be tailored to the specific setting of implementation, and empirical research is needed to confirm the conclusions.


Management of Amelogenesis Imperfecta: A 15-Year Case History of Two Siblings.
Amelogenesis imperfecta (AI) is a heterogenous genetic disorder that interferes with normal enamel formation in the absence of systemic disorders. The patients' main concerns are caries susceptibility, poor esthetics, and generalized sensitivity. There is a broad clinical spectrum, from discolorations to consequent enamel alterations. This case report describes the 15-year case study and the full-mouth rehabilitation of two siblings affected by a hypocalcified AI. Clinical Considerations: In these two patients, conservative care with stainless steel crowns and direct composite restorations was undertaken to restore function and esthetics and to reduce sensitivities in primary and mixed dentitions. The difficulties in monitoring resulted in severe infectious complications (dental abscess with cutaneous fistula), important dental defects, and loss of spaces with subsequent malocclusion. In the young adult dentition, they were treated by extractions, root canal therapies, and new restorations: stainless steel crowns for permanent molars, direct composite restorations (with strip crowns) for incisors and maxillary canines (to improve the crown morphology as well as to mask the discolorations and the malpositions), and adjusted composite crown molds using a thermoforming procedure for premolars and the mandibular canines. The main difficulties were rapid tooth surface loss, bonding to atypical enamel, developing dentition, long-term follow-up. Restoring function and esthetics in AI-affected patients is a challenge from primary to adult dentition. Early corrections are essential to avoid dental damage and for psychological benefits. This clinical report highlights the adhesive rehabilitation for anterior and premolar areas and the difficulty of patient follow-up.


Congenital transcobalamin II deficiency due to errors in RNA editing.
Transcobalamin II (TCII) is a plasma protein essential for the transport and cellular uptake of vitamin B12 (B12; cobalamin, Cbl). Congenital deficiency of functional TCII is an autosomal recessive genetic disorder that results in clinical B12 deficiency usually within several months following birth. In this report, we describe the molecular basis for TCII deficiency in two patients who developed a megaloblastic anemia in early infancy. The serum of both patients contained immunoreactive TCII that did not bind [57Co]Cbl. The fibroblasts from each patient secreted a similarly nonfunctional TCII, yet full-length TCII transcripts were identified by Northern blot. Overlapping cDNA fragments were generated by reverse transcription-polymerase chain reaction and several mutations were identified in the coding region of the cDNA, one of which was common to both patients. However, amplification of the corresponding regions of the gene from genomic DNA failed to identify these mutations. These findings were confirmed by replicate analyses and support the proposal that a variance in RNA editing is the likely mechanism for the mutations that resulted in the expression of a nonfunctional TCII protein in these patients.


Root and canal morphology of Thai maxillary molars.
To investigate the root and canal morphology of 268 maxillary permanent molars collected from an indigenous Thai population. The cleaned teeth were accessed, the pulp dissolved by sodium hypochlorite under ultrasonication, and the pulp system injected with Indian ink. The teeth were rendered clear by demineralization and immersion in methyl salicylate. The following observations were made: (i) number of roots and their morphology; (ii) number of root canals per root; (iii) root-canal configuration in each root using Vertucci's classification with additional modifications; and (iv) presence and location of lateral canals and intercanal communications. All the maxillary first and second molars had three separate roots. Only, half (51%) of the maxillary third molars had three separate roots; the other half had fused or conical roots. The majority of the distobuccal (98.1-100%) and palatal (100%) roots had type I canals. Over half of the mesiobuccal roots of first (65%) and second (55%) molars had two canals. The most common (44.2%) canal configuration in mesiobuccal roots of first molars was type IV (two canals, two foramina). A variety of canal types were found in the mesiobuccal roots of second molars. Maxillary third molars showed the greatest diversity of canal morphology. There was an increase in the prevalence of lateral canals towards the apical part of the roots and intercanal communications were present in 16% of each of first, second and third Thai maxillary molars. The mesiobuccal roots of Thai maxillary molars possessed a variety of canal system types. Over 50% of the first molars had a second mesiobuccal canal. The palatal and distobuccal canals mainly had type I canals. Only, a small proportion (7.3-13.3%) of the roots exhibited lateral canals which were the most common in the apical third


State variations in home health expenditures and utilization under Medicare and Medicaid.
Expenditures for home health services provided under Medicare and Medicaid nearly tripled between 1977 and 1982. Utilization and expenditures for home health services vary widely across the states for both programs, and this article examines in some detail the nature and extent of these variations. States vary considerably with respect to relative utilization, visits, and visit-expenditures under Medicare; and states vary in terms the role visits versus visit-reimbursements play in shaping overall costs. The Medicaid program is effectively two programs: New York; and the other states, with eight accounting for half of Medicaid expenditures outside New York.


Approximating dynamic proximity with a hybrid geometry energy-based kernel for diffusion maps.
The diffusion map is a dimensionality reduction method. The reduction coordinates are associated with the leading eigenfunctions of the backward Fokker-Planck operator, providing a dynamic meaning for these coordinates. One of the key factors that affect the accuracy of diffusion map embedding is the dynamic measure implemented in the Gaussian kernel. A common practice in diffusion map study of molecular systems is to approximate dynamic proximity with RMSD (root-mean-square deviation). In this paper, we present a hybrid geometry-energy based kernel. Since high energy-barriers may exist between geometrically similar conformations, taking both RMSD and energy difference into account in the kernel can better describe conformational transitions between neighboring conformations and lead to accurate embedding. We applied our diffusion map method to the β-hairpin of the B1 domain of streptococcal protein G and to Trp-cage. Our results in β-hairpin show that the diffusion map embedding achieves better results with the hybrid kernel than that with the RMSD-based kernel in terms of free energy landscape characterization and a new correlation measure between the cluster center Euclidean distances in the reduced-dimension space and the reciprocals of the total net flow between these clusters. In addition, our diffusion map analysis of the ultralong molecular dynamics trajectory of Trp-cage has provided a unified view of its folding mechanism. These promising results demonstrate the effectiveness of our diffusion map approach in the analysis of the dynamics and thermodynamics of molecular systems. The hybrid geometry-energy criterion could be also useful as a general dynamic measure for other purposes.


Baicalin Alleviates LPS-Induced Oxidative Stress via NF-κB and Nrf2-HO1 Signaling Pathways in IPEC-J2 Cells.
Baicalin is a natural plant extract with anti-inflammatory and anti-oxidant activities. However, the molecular mechanism of baicalin on oxidative stress in IPEC-J2 cells exposed to LPS remains to be unclear. In this study, LPS stimulation significantly increased Toll-like receptor 4, tumor necrosis factor-α, and interleukins (IL-6 and IL-1β) expression in IPEC-J2 cells, and it activated the nuclear factor (NF-κB) expression. While, baicalin exerted anti-inflammatory effects by inhibiting NF-κB signaling pathway. LPS stimulation significantly increased the levels of the oxidative stress marker MDA, inhibited the anti-oxidant enzymes catalase and superoxide dismutase, which were all reversed by baicalin pre-treatment. It was found that baicalin treatment activated the nuclear import of nuclear factor-erythroid 2 related factor 2 (Nrf2) protein, and significantly increased the mRNA and protein expression of its downstream anti-oxidant factors such as heme oxygenase-1 and quinone oxidoreductase-1, which suggested that baicalin exerted anti-oxidant effects by activating the Nrf2-HO1 signaling pathway. Thus, pretreatment with baicalin inhibited LPS - induced oxidative stress and protected the normal physiological function of IPEC-J2 cells via NF-κB and Nrf2-HO1 signaling pathways.


[Bulimia nervosa--therapeutic possibilities with emphasis on cognitive behavioral therapy].
The author pays attention to recent trends in the treatment of bulimia nervosa, he discusses problems of therapeutic methods and hospital treatment. Special attention is paid to ambulatory and cognitive behavioural therapy. The author emphasizes its integrating character due to the complexity of the problem of eating disorders. The latter are becoming one of the most serious health and social problems of adolescent women not only in western Europe and the USA but also in this country. The author defines some principles of the cognitive behavioural approach in ambulatory treatment of bulimia which is briefly described in the conclusion of the article. He reminds that it is always essential to take into account the grade of the disorder, the differential character of its psychosocial picture and the patient's needs. This should be matched by the offered therapeutic methods using different possibilities of in- and out-patient treatment. An essential prerequisite of treatment is not only detailed knowledge of the therapeutic method but also of problems associated with impaired food intake.


Acute Lumbar Paraspinal Compartment Syndrome After Strenuous Exercise: A Case Report.
Acute lumbar paraspinal compartment syndrome (APCS) is rare. This case report describes the clinical presentation and magnetic resonance imaging (MRI) findings of APCS in a young man after strenuous weight training. The patient underwent surgical decompression of the paraspinal muscles with fasciotomy 4 days after presentation and made a complete functional recovery at the 3-month follow-up postsurgery. This case report highlights the importance of MRI in facilitating the diagnosis of APCS and demonstrates that fasciotomy, even if delayed, can result in complete functional recovery. This supports current understanding that surgical fascial decompression is the preferred treatment over conservative measures for this condition.


How to design effective vaccines: lessons from an old success story.
Evaluation of: Gaucher D, Therrien R, Kettaf N et al. Yellow fever vaccine induces integrated multilineage and polyfunctional immune responses. J. Exp. Med. 205(13), 3119-3131 (2008). Despite the successful development of vaccines that are able to elicit potent and protective immune responses, the majority of vaccines were developed empirically and the mechanistic events leading to protective immune responses are often poorly understood. This impedes the development of new prophylactic as well as therapeutic vaccines for infectious diseases and cancer. Gaucher et al. took advantage of the effective yellow fever vaccine 17D to prospectively identify key immunological responses elicited by the vaccine using functional genomics and flow cytometric analysis. The results of the study clearly indicate 'that the immune response to a strong vaccine is preceded by the coordinated induction of master transcription factors that lead to the development of a broad, polyfunctional and persistent immune response integrating all effector cells of the immune system'.


A review of health utilities using the EQ-5D in studies of cardiovascular disease.
The EQ-5D has been extensively used to assess patient utility in trials of new treatments within the cardiovascular field. The aims of this study were to review evidence of the validity and reliability of the EQ-5D, and to summarise utility scores based on the use of the EQ-5D in clinical trials and in studies of patients with cardiovascular disease. A structured literature search was conducted using keywords related to cardiovascular disease and EQ-5D. Original research studies of patients with cardiovascular disease that reported EQ-5D results and its measurement properties were included. Of 147 identified papers, 66 met the selection criteria, with 10 studies reporting evidence on validity or reliability and 60 reporting EQ-5D responses (VAS or self-classification). Mean EQ-5D index-based scores ranged from 0.24 (SD 0.39) to 0.90 (SD 0.16), while VAS scores ranged from 37 (SD 21) to 89 (no SD reported). Stratification of EQ-5D index scores by disease severity revealed that scores decreased from a mean of 0.78 (SD 0.18) to 0.51 (SD 0.21) for mild to severe disease in heart failure patients and from 0.80 (SD 0.05) to 0.45 (SD 0.22) for mild to severe disease in angina patients. The published evidence generally supports the validity and reliability of the EQ-5D as an outcome measure within the cardiovascular area. This review provides utility estimates across a range of cardiovascular subgroups and treatments that may be useful for future modelling of utilities and QALYs in economic evaluations within the cardiovascular area.


Robust Assessment of Molecular Size Distribution of Meningococcal AC Vaccine.
The molecular size of meningococcal polysaccharides is an important physicochemical parameter that is related to immunogenicity and efficacy. A simple method for size-exclusion chromatography was developed, optimized, and applied for safe and rapid fractionation of meningococcal polysaccharide AC vaccine. Pooling of the fractions collected from size-exclusion chromatography was investigated and evaluated, rather than analyzing each fraction separately, for determining the percentages of meningococcal polysaccharide A and C that were eluted before the distribution coefficient of 0.5. Pooling is preferred rather than analyzing each fraction individually, as it is easily handled, faster, simpler, less expensive, more accurate, safe, and applicable. The developed method was validated and successfully applied for the determination of meningococcal polysaccharide vaccine serotype A and C in quality-control and commercial samples.


Biochemical and biological characterization of a new oxidized avidin with enhanced tissue binding properties.
Chicken avidin and bacterial streptavidin are widely employed in vitro for their capacity to bind biotin, but their pharmacokinetics and immunological properties are not always optimal, thereby limiting their use in medical treatments. Here we investigate the biochemical and biological properties of a new modified avidin, obtained by ligand-assisted sodium periodate oxidation of avidin. This method allows protection of biotin-binding sites of avidin from inactivation caused by the oxidation step and delay of avidin clearance from injected tissue by generation of aldehyde groups from avidin carbohydrate moieties. Oxidized avidin shows spectroscopic properties similar to that of native avidin, indicating that tryptophan residues are spared from oxidation damage. In strict agreement with these results, circular dichroism and isothermal titration calorimetry analyses confirm that the ligand-assisted oxidation preserves the avidin protein structure and its biotin binding capacity. In vitro cell binding and in vivo tissue residence experiments demonstrate that aldehyde groups provide oxidized avidin the property to bind cellular and interstitial protein amino groups through Schiff's base formation, resulting in a tissue half-life of 2 weeks, compared with 2 h of native avidin. In addition, the efficient uptake of the intravenously injected (111)In-BiotinDOTA (ST2210) in the site previously treated with modified avidin underlines that tissue-bound oxidized avidin retains its biotin binding capacity in vivo. The results presented here indicate that oxidized avidin could be employed to create a stable artificial receptor in diseased tissues for the targeting of biotinylated therapeutics.


Transforming growth factor-beta increases elastin production by neonatal rat lung fibroblasts.
While elastin degradation is a hallmark of pulmonary emphysema, it is likely that elastin synthesis also occurs. However, the supramolecular structure and function of the newly synthesized elastin are abnormal. Very little is known about the regulation of elastin synthesis during the development of emphysema when prominent collections of mononuclear phagocytes are found in and near the alveolar interstitium. Transforming growth factor-beta (TGF-beta) is an important regulator of collagen and fibronectin production in wound healing, which is also accompanied by an influx of mononuclear phagocytes. We hypothesized that TGF-beta may influence elastin production by fibroblasts in the pulmonary interstitium. Therefore, we examined the influence of TGF-beta on the production of elastin by postconfluent cultures of neonatal rat lung fibroblasts. Elastin production was quantitated by analyzing the incorporation of [3H]valine into the soluble elastin precursor tropoelastin (TE). The incorporation of [3H]valine into TE was approximately 2-fold greater in the presence of 40 or 100 pM TGF-beta than in its absence. The intracellular, free [3H]valine pool was increased by 18% in the presence of TGF-beta. Therefore, TGF-beta-related differences in the precursor pool size were not solely responsible for the observed increase in [3H]valine incorporation. Northern analysis demonstrated that the increase in TE was accompanied by a smaller but significant increase in the steady-state level of elastin mRNA. Thus, the observed increase in TE production can be at least partially attributed to a pretranslational effect of TGF-beta.


The role of single-channel nasal airflow pressure transducer in the diagnosis of OSA in the sleep laboratory.
Obstructive sleep apnea (OSA) is a common but underdiagnosed disorder. There is a need for validated simpler modalities such as single-channel monitors to assist diagnosis of OSA. To assess data sufficiency, agreement, and diagnostic accuracy of nasal airflow measured by a single-channel pressure transducer device (Flow Wizard, DiagnoseIT, Sydney, Australia) compared to attended full polysomnography (PSG) on the same night for OSA diagnosis. Cross-sectional study. Laboratory. Subjects with possible OSA referred to the sleep laboratory for PSG were eligible. Nasal airflow was measured by a pressure transducer in the laboratory concurrently with PSG. Of 226 eligible subjects who consented, 221 (97.8%; 151 males, 70 females) completed the protocol. With nasal airflow measurement, 5.3% of subjects had insufficient data, compared with 2.2% on PSG. The mean difference between PSG AHI and NF RDI was -6.2 events/h with limits of agreement (+/- 2 standard deviation [SD]) of 17.0 events/hr. The accuracy of the Flow Wizard for diagnosing severe OSA (PSG AHI > 30) was very good (area under the ROC curve [AUC] 0.96; 95% confidence interval [CI] 0.92 to 0.99) and for diagnosing OSA (PSG AHI > 5) was good (AUC, 0.84; 95% CI, 0.77 to 0.90). There was no difference in the rate of data insufficiency and accuracy between males and females. Nasal flow measured by a nasal pressure transducer has a low rate of data insufficiency, good agreement, and high accuracy compared to PSG for diagnosing OSA in the monitored sleep laboratory setting.


SAM levels, gene expression of SAM synthetase, methionine synthase and ACC oxidase, and ethylene emission from N. suaveolens flowers.
S'adenosyl-L: -methionine (SAM) is a ubiquitous methyl donor and a precursor in the biosynthesis of ethylene, polyamines, biotin, and nicotianamine in plants. Only limited information is available regarding its synthesis (SAM cycle) and its concentrations in plant tissues. The SAM concentrations in flowers of Nicotiana suaveolens were determined during day/night cycles and found to fluctuate rhythmically between 10 and 50 nmol g(-1) fresh weight. Troughs of SAM levels were measured in the evening and night, which corresponds to the time when the major floral scent compound, methyl benzoate, is synthesized by a SAM dependent methyltransferase (NsBSMT) and when this enzyme possesses its highest activity. The SAM synthetase (NsSAMS1) and methionine synthase (NsMS1) are enzymes, among others, which are involved in the synthesis and regeneration of SAM. Respective genes were isolated from a N. suaveolens petal cDNA library. Transcript accumulation patterns of both SAM regenerating enzymes matched perfectly those of the bifunctional NsBSMT; maximum mRNA accumulations of NsMS1 and NsSAMS1 were attained in the evening. Ethylene, which is synthesized from SAM, reached only low levels of 1-2 ppbv in N. suaveolens flowers. It is emitted in a burst at the end of the life span of the flowers, which correlates with the increased expression of the 1-aminocyclopropane-1-carboxylate oxidase (NsACO).


From the Cover: Catalytic Antioxidant Rescue of Inhaled Sulfur Mustard Toxicity.
Sulfur mustard (bis 2-chloroethyl ethyl sulfide, SM) is a powerful bi-functional vesicating chemical warfare agent. SM tissue injury is partially mediated by the overproduction of reactive oxygen species resulting in oxidative stress. We hypothesized that using a catalytic antioxidant (AEOL 10150) to alleviate oxidative stress and secondary inflammation following exposure to SM would attenuate the toxic effects of SM inhalation. Adult male rats were intubated and exposed to SM (1.4 mg/kg), a dose that produces an LD50 at approximately 24 h. Rats were randomized and treated via subcutaneous injection with either sterile PBS or AEOL 10150 (5 mg/kg, sc, every 4 h) beginning 1 h post-SM exposure. Rats were euthanized between 6 and 48 h after exposure to SM and survival and markers of injury were determined. Catalytic antioxidant treatment improved survival after SM inhalation in a dose-dependent manner, up to 52% over SM PBS at 48 h post-exposure. This improvement was sustained for at least 72 h after SM exposure when treatments were stopped after 48 h. Non-invasive monitoring throughout the duration of the studies also revealed blood oxygen saturations were improved by 10% and clinical scores were reduced by 57% after SM exposure in the catalytic antioxidant treatment group. Tissue analysis showed catalytic antioxidant therapy was able to decrease airway cast formation by 69% at 48 h post-exposure. To investigate antioxidant induced changes at the peak of injury, several biomarkers of oxidative stress and inflammation were evaluated at 24 h post-exposure. AEOL 10150 attenuated SM-mediated lung lipid oxidation, nitrosative stress and many proinflammatory cytokines. The findings indicate that catalytic antioxidants may be useful medical countermeasure against inhaled SM exposure.


[Proteins of bacterial membranes. Purification of soluble ATPase from Acholeplasma laidlawii].
A purified preparation of ATPase (factor F1) from the Acholeplasma laidlawii was obtained. The purification procedure included extraction of the enzyme complex from the isolated membranes by ultrasonication, chromatography on DEAE-cellulose and gel filtration on Sepharose 6B. The specific activity of the ATPase was increased 30-fold as compared to the original activity. The Km value for ATP hydrolysis was 7,4 . 10(-4) M. ADP competitively inhibited the enzyme (Ki = 2,0 . 10(-4) M). Ouabain (2,5 . 10(-4) M) and dicyclohexylcarbodiimide (1,0 . 10(-4) M) did not inhibit the ATPase activity. The enzyme was activated by Mg2+, but was inhibited by a combination of Na+ and K+. The enzyme is cold-labile, but can be stabilized by storage in buffer solutions, containing methanol, glycerol or lecithin.


Towards A Deeper Understanding of the Interfacial Adsorption of Enzyme Molecules in Gigaporous Polymeric Microspheres.
Compared with the one immobilized in the conventional mesoporous microspheres, the enzyme immobilized in gigaporous microspheres showed much higher activity and better stability. To gain a deeper understanding, we herein selected lipase as a prototype to comparatively analyze the adsorption behavior of lipase at interfaces in gigaporous and mesoporous polystyrene microspheres at very low lipase concentration, and further compared with the adsorption on a completely flat surface (a chip). Owing to the limited space of narrow pores, lipase molecules were inclined to be adsorbed as a monolayer in mesoporous microspheres. During this process, the interaction between lipase molecules and the interface was stronger, which could result in the structural change of lipase molecular and compromised specific activity. In addition to monolayer adsorption, more multilayer adsorption of enzyme molecules also occurred in gigaporous microspheres. Besides the adsorption state, the pore curvature also affected the lipase adsorption. Due to the multilayer adsorption, the excellent mass transfer properties for the substrate and the product in the large pores, and the small pore curvature, lipase immobilized in gigaporous microspheres showed better behaviors.


Sealing of adhesive systems in ferric sulfate-contaminated dentinal margins in class V composite resin restorations.
Background. Hemostatic agents are applied to prepare an isolated bleeding-free condition during dental treatments and can influence adhesive restorations. This study evaluated the effect of a hemostatic agent (ViscoStat) on microleakage of contaminated dentinal margin of class V composite resin restorations with three adhesives. Methods. Sixty freshly extracted human molars were selected and class V cavities (3×3×1.5 mm) were prepared on buccal and lingual surfaces. Gingival margins of the cavities were placed below the cementoenamel junction. The teeth were divided into six groups randomly. The adhesives were Excite, AdheSE and AdheSE One. In three groups, the gingival walls of the cavities were contaminated with ViscoStat and then rinsed. The cavities were restored with composite resin and light-cured. After storage in distilled water (37°C) for 24 hours and polishing, the samples were thermocycled and sealed with nail varnish. Then they were stored in 1% basic fuchsin for 24 hours, rinsed and mounted in self-cured acryl resin, followed by sectioning buccolingually. Dye penetration was observed under a stereomicroscope and scored. Data were statistically analyzed with Kruskal-Wallis and Mann-Whitney U tests. P<0.05 was set as the level of significance. Results. Only in the Excite group, contamination did not have adverse effects on dentin microleakage (P > 0.05). In the contaminated groups, Excite had significantly less microleakage than the others (P = 0.003). AdheSE and AdheSE One did not exhibit significant difference in microleakage (P > 0.05). Conclusion. ViscoStat hemostatic agent increased dentinal microleakage in AdheSE and AdheSE One adhesives with no effect on Excite.


Skin3D: Detection and longitudinal tracking of pigmented skin lesions in 3D total-body textured meshes.
We present an automated approach to detect and longitudinally track skin lesions on 3D total-body skin surface scans. The acquired 3D mesh of the subject is unwrapped to a 2D texture image, where a trained objected detection model, Faster R-CNN, localizes the lesions within the 2D domain. These detected skin lesions are mapped back to the 3D surface of the subject and, for subjects imaged multiple times, we construct a graph-based matching procedure to longitudinally track lesions that considers the anatomical correspondences among pairs of meshes and the geodesic proximity of corresponding lesions and the inter-lesion geodesic distances. We evaluated the proposed approach using 3DBodyTex, a publicly available dataset composed of 3D scans imaging the coloured skin (textured meshes) of 200 human subjects. We manually annotated locations that appeared to the human eye to contain a pigmented skin lesion as well as tracked a subset of lesions occurring on the same subject imaged in different poses. Our results, when compared to three human annotators, suggest that the trained Faster R-CNN detects lesions at a similar performance level as the human annotators. Our lesion tracking algorithm achieves an average matching accuracy of 88% on a set of detected corresponding pairs of prominent lesions of subjects imaged in different poses, and an average longitudinal accuracy of 71% when encompassing additional errors due to lesion detection. As there currently is no other large-scale publicly available dataset of 3D total-body skin lesions, we publicly release over 25,000 3DBodyTex manual annotations, which we hope will further research on total-body skin lesion analysis.


Modulation of cytotoxicity and cellular pharmacology of 1,2-diaminocyclohexane platinum (IV) complexes mediated by axial and equatorial ligands.
Isomers (R,R-, S,S-, and cis-) of 1,2-diaminocyclohexane (DACH) platinum(IV) complexes with selected axial and equatorial ligands were synthesized and evaluated for in vitro antitumor activity, cellular uptake, and total DNA-Pt adducts. L1210 cells, sensitive to cis-diamminedichloroplatinum(II) (CDDP) and tetraplatin (L1210/0), 160-fold resistant to CDDP [L1210/diamminedichloroplatinum (DDP)], or 70-fold resistant to tetraplatin (L1210/DACH), were used in conjunction with compounds having the general structure DACH-Pt(IV)-X2Y2, where X and Y are axial and equatorial ligands and X2Y2 are specifically Cl2Cl2,Ac2Cl2, (TFA)2Cl2, (OH)2Cl2, and Cl2CBDCA (Cl, chloro; Ac, acetato; TFA, trifluoroacetato; OH, hydroxo; CBDCA, 1,1-cyclobutanedicarboxylato). Comparison of cytotoxicities between isomers of Cl2Cl2,Ac2Cl2, or Cl2CBDCA indicated that R,R-isomers were the most effective against all three cell lines. The relatively lower activity of the S,S- and cis-isomers was cell line dependent: against L1210/DACH, both isomers of Cl2Cl2 were only 2- to 3-fold less effective, and this contrasted with 7- and 26-fold lower cytotoxicities, respectively, against L1210/DDP. Cross-resistance factors in the L1210/DDP and L1210/DACH lines depended on both isomeric form and the nature of axial or equatorial ligand. The L1210/DDP cells were 6- to 9-fold cross-resistant to the R,R-isomer, 8- to 15-fold to S,S-isomer, and 13- to 38-fold to cis-isomer. The L1210/DACH line was only 4- to 7-fold cross-resistant to the three isomers of Ac2Cl2 but cross-resistance to the isomers was 47- to 79-fold for Cl2Cl2 and 22- to 56-fold for Cl2CBDCA complexes. Compared with CDDP, accumulation (2 h at 100 microM drug concentration) of Ac2Cl2 in the three L1210 cell lines was 26-50%, while uptake of Cl2Cl2 and (TFA)2Cl2 was 100-170% and 320-570%, respectively. The greatest DNA binding was seen with Cl2Cl2 in all cell lines, followed by (TFA)2Cl2, CDDP, and Ac2Cl2. DNA binding correlated directly with potency (1/concentration producing 50% inhibition) in the L1210/0 model (r = 0.973, P < 0.016) but not in the L1210/DDP and L1210/DACH models. Accumulation and DNA-binding studies indicated that binding efficiency to DNA was: Cl2Cl2 > Ac2Cl2 > CDDP > (TFA)2Cl2. In a nonreducing environment, the Pt(IV) complexes (20 microM) did not react with salmon sperm DNA. Reduced glutathione (100 microM), as a reducing agent, rendered full binding capacity to Cl2Cl2; binding was 25-30% of the expected maximum for (TFA)2Cl2, while Ac2Cl2 remained inert.(ABSTRACT TRUNCATED AT 400 WORDS)


Increased pro-opiomelanocortin-derived peptide release in myotonic dystrophy.
The response of plasma immunoreactive (IR)-ACTH, IR-beta-endorphin (beta-END) and IR-cortisol to insulin-induced hypoglycaemia, an acute stimulus to the pituitary corticotrophs through the central nervous system, and to synthetic ovine corticotrophin-releasing hormone (CRH), a direct corticotroph stimulator, were studied in normal males and males with myotonic dystrophy. Myotonics had an increased IR-ACTH and IR-beta-END response to hypoglycaemia and an increased IR-ACTH response to CRH compared with normals. Plasma IR-cortisol response were not different in either group of subjects to both stimuli. This neuroendocrine abnormality in myotonic dystrophy may represent a manifestation of the purported specific cell membrane defect underlying the disease. This is the first report of an abnormality in proopiomelanocortin peptide release in myotonic dystrophy.


Semen sample collection in medium enhances the implantation rate following ICSI in patients with severe oligoasthenoteratozoospermia.
Ejaculation in medium increases the proportion of antibody-free spermatozoa in semen samples containing anti-sperm antibodies and thereby enhances the fertilization rate in vitro. The aim of this study was to investigate whether this technique is also beneficial in semen samples with severe oligoasthenoteratozoospermia (OAT) where bacteria and detritus are often present. A prospective randomized controlled trial was carried out to study the results of sperm preparation and fertilization and pregnancy rates after intracytoplasmic sperm injection (ICSI) for OAT. Of the 114 couples (one cycle per couple) studied between 1998 and 2000, 55 men were randomized to have semen collection into sterile dry pots (group A) and the remaining 59 had samples collected into 20 ml HEPES buffered Ham's F-10 medium with 10% human serum albumin (group B). In group B the ejaculates were incubated for 30 min and mixed gently. The samples were then processed by mild centrifugation and washing followed by a mini-swim-up technique. The ejaculates in group A were prepared by the swim-up procedure only. The overall fertilization rate was 71.8% and was similar in groups A (fertilization rate = 66.7%) and B (fertilization rate = 64.3%). In group A, 10/55 clinical pregnancies were recorded (pregnancy rate 18%), with an implantation rate (IR) of 6.9% per embryo. In group B, 16 of 59 patients conceived leading to significantly higher implantation (9.9%, P < 0.001) and clinical pregnancy rates (27%, P < 0.001). It is postulated that the addition of medium before liquefaction could inhibit the binding of bacteria and detritus to the sperm surface and may diminish DNA damage caused by reactive oxygen species, leading to improved efficiency of fertilization. The results demonstrate that the addition of HEPES buffered Ham's F-10 medium to sample collection pots significantly improves the pregnancy rate after ICSI in patients with severe OAT.


Identifying sequences potentially related to resistance response of Piper tuberculatum to Fusarium solani f. sp. piperis by suppression subtractive hybridization.
Piper tuberculatum is an exotic Piper from the Amazon region that shows resistance to infection by Fusarium solani f. sp. piperis, causal agent of Fusarium disease in black pepper (Piper nigrum L.). In this work we aimed to study the interaction between P. tuberculatum and F. solani f. sp. piperis at a molecular level, using suppression subtractive hybridization to identify genes potentially related to Fusarium disease resistance. Comparative sequence analysis confirmed that clones isolated here show a high identity with genes coding for proteins that have a known role in plant defense response mechanisms, such as peroxidase, hydroxyproline-rich glycoprotein and CBL-interacting protein kinase. The present study constitutes the first effort to understand the molecular basis of this plant-pathogen interaction, identifying genes which may be used in the future genetic improvement of black pepper.