We describe a patient with an implanted pacemaker for impaired AV conduction who presented with  an incessant tachycardia. EP study showed that the tachycardia was atrioventricular nodal reentrant tachycardia (AVNRT) with repeated spontaneous initiation because of poor or absent antegrade fast pathway conduction. Slow pathway ablation was successful in terminating the tachycardia and making it non-inducible. During a follow-up visit in the pacemaker clinic, a 45-year-old male complained of persistent palpitations for the past 2 months. He had undergone implantation of a permanent single chamber pacemaker with a ventricular lead 18 years back for trifascicular block with intermittent complete heart block and had subsequently undergone pulse generator replacement twice. His heart rate in the clinic was 130 bpm. Echocardiogram showed normal LV function without any abnormalities. He was not on any drugs. Electrocardiogram showed a broad complex tachycardia with a right bundle branch block (RBBB) morphology, 1:1 ventriculo-atrial (VA) association and a short RP interval. An electrophysiology study was planned in view of the persistent tachycardia.In the electrophysiology lab, he had tachycardia at the beginning of the procedure. The tachycardia could be easily terminated with ventricular overdrive pacing, and on termination of the tachycardia he was found to have sinus rhythm with a PR interval of 380 ms and broad QRS of RBBB morphology. However, tachycardia would always reinitiate spontaneously after a few beats of sinus rhythm . During Intracardiac electrograms during tachycardia showed 1:1 VA association with VA interval of 95 ms and central atrial activation. RV overdrive pacing at a cycle length (CL) slightly shorter than the tachycardia CL showed a ventricle-atrium-ventricle (VAV) response on termination of pacing with post pacing interval 216 ms longer than the tachycardia cycle length and VA interval was 150 ms longer than stimulus-atrial (SA) interval, all consistent with typical slow-fast atrioventricular nodal reentrant tachycardia (AVNRT) .Due to the incessant nature of the tachycardia, mapping had to be performed in tachycardia in the region of the slow pathway guided by fluoroscopy and electrograms. Ablation near the os of the coronary sinus was accompained by junctional beats. Post ablation, patient remained in sinus rhythm. AH and HV intervals were 228 ms and 50 ms respectively. Atrial programmed stimulation showed no dual pathway physiology and tachycardia could not be induced with atrial premature stimuli or burst pacing.AVNRT usually presents as a paroxysmal form of tachycardia with each episode lasting for a short period of time. We could find only one report of incessant presentation of AVNRT . In pati


IL-4 production was increased by the pretreatment whereas synthesis ofIFN-γ was diminished. Addition of DEX in the second activation suppressed all cytokineproduction. In brief, the transient presence of glucocorticoids in the culture induces a change in the pattern ofcytokine production but the continuous presence causes inhibition of cytokine synthesis. Further studies in whichIL-4 was used together with DEX showed that the cytokine potentiated the effect of the hormone.Purified rat CD4            The data here presented suggest that glucocorticoids and the neuroendocrine system may be expectedto have long-term immunological effects as well as short-lived immunosuppressive ones. High concentration ofglucocorticoids suppress cytokine production but when steroids return to basal levels the immune response isdirected in a way that favors Th2-type reactions. Possible implications regarding the immune response to pathogensand autoantigens are discussed.


Giant ascending aortic aneurysm formation following aortic valve replacement is rare. A 28-year-old man who underwent aortic valve replacement with a prosthetic valve for aortic regurgitation secondary to congenital bicuspid aortic valve about 10 years ago was diagnosed with a giant ascending aortic aneurysm about 16 cm in diameter in follow-up. The aneurysm was resected leaving the functional old mechanical prosthesis in place and implanted a 34-mm Hemashield woven graft, associated with the left and right coronary artery button implantation. Histological findings of the aortic aneurysm wall showed cystic medial necrosis. The postoperative course was uneventful and postoperative examination demonstrated good surgical results. Patients with bicuspid aortic valve are at increased risk for aortic complications and that aortic valve replacement does not prevent progressive aortic dilatation , 2. In tA 28-year-old man with severe aortic regurgitation underwent aortic valve replacement with a mechanical prosthetic valve about 10 years ago. At the time of this operation the ascending aorta was slightly dilated and measured as 3.4 cm. An ascending aortic aneurysm was suspected with chest X-ray in routine follow-up of the patient who had not been controlled until that time . EchocarAortic complications occurring after aortic valve replacement (AVR) include aortic dissection, ascending aortic aneurysm, aortic root aneurysm, and pseudoaneurysm –4. To ouThe presented case had been operated before for severe aortic regurgitation secondary to congenital bicuspid aortic valve. Bicuspid aortic valve (BAV) is one of the most common congenital disorders involving the heart valves, with a prevalence ranging from 1% to 2% of the entire population , 5 demonBased on the law of Laplace, wall tension increases as the radius of an aneurysm increases (tension = pressure × radius). It is therefore intuitive that larger aneurysms have a greater risk of rupture. Coady et al.  have wriWe believe that the patients with congenital BAV should be followed regularly in terms of aortic dilatation and aneurysm formation even if their aortic valve is replaced. The surgical management of aortic aneurysm should be done in order to decrease the morbidity and mortality in those patients before the diameter of aneurysm not exceeding relatively larger sizes.


Scattering of shorter-wavelength visible light limits thefluorescence imaging depth of thick specimens such as wholeorgans. In this study, we report the use of four newly synthesizednear-infrared and far-red fluorescence probesto image tumor cells in the subpleural vasculature of the intactrat lungs. Transpelural imaging of tumor cells labeled withlong-wavelength probes and expressing green fluorescent protein was done in the intact ratlung after perfusate administration or intravenous injection. Ourresults show that the average optimum imaging depth for thelong-wavelength probes is higher (27.8 ± 0.7 μm) than for GFP , corresponding to a40% increase in the volume of tissue accessible forhigh-resolution imaging. The maximum depth of cell visualizationwas significantly improved with the novel dyes  compared with GFP . Stable binding of the long-wavelengthvital dyes to the plasma membrane also permitted in vivo trackingof injected tumor cells in the pulmonary vasculature. These probesoffer a significant improvement in the imaging quality of in situbiological processes in the deeper regions of intact lungs.


UK CropNet currently provides a range of databases (and database-mining tools) to the plant community that are all freely accessible through our website (http://ukcrop.net/). Recent upgrades have meant that we can now expand the range of available facilities (e.g. addition of new databases) whilst also strengthening and improving access to existing services (e.g. providing a BLAST search facility against sequences in our databases). This article will briefly outline these and other new developments in our service.


Gene expression similarity measuring methods were developed and applied to search rapidly growing public microarray databases. However, current expression similarity measuring methods need to be improved to accurately measure similarity between gene expression profiles from different platforms or different experiments.We devised new gene expression similarity measuring method based on pathway information. In short, newly devised method measure similarity between gene expression profiles after converting them into pathway based expression profiles. To evaluate pathway based gene expression similarity measuring method, we conducted cell type classification test. Pathway based similarity measuring method shows higher classification accuracy. Especially, pathway based methods outperform at most 50% and 10% over conventional gene expression similarity method when search databases are limited to cross-platform profiles and cross-experiment profiles.The pathway based gene expression similarity measuring method outperforms commonly used similarity measuring methods. Considering the fact that public microarray database is consist of gene expression profiles of various experiments with various type of platform, pathway based gene expression similarity measuring method could be successfully applied for searching large public microarray databases. As microarray experiment has been widely used for various field of biology, public microarray databases have been rapidly growing each year. Currently, the two largest microarray databases, GEO  and ArraIn accordance with this expensive collection of large scale gene expression databases, database searching methods have been developed to make the database easily accessible and practically useful. Since the microarray data is deposited in public microarray database as unit of experiment which is consist of several individual gene expression profiles, search methods also have evolved into two way, experiment dataset level search and individual gene expression profile level search.Most of experiment dataset level search methods are depend on dataset annotation by authors of dataset. Atul B. et al. has been tried to classify the gene expression experiment dataset in GEO (GEO series) by annotating each GEO dataset with medical language terms such as UMLS and SNOMED -5 and toAlong the attempts to search large public microarray databases at experiment dataset level, individual gene expression profile level search method has also been conceptualized and developed . GEST 99 is the Here we try to improve similar gene expression profile search. For this purpose, we devised a pathway based gene expression similarity measuring method. Our pathway based methods outperform conventional method especially for cross-platform and cross-experiment profile search.We used set of gene expression profiles curated by CellMontage group . Each geFor the classification procedure, we first selected cell types with which at least two different platform types are associated. For each selected cell type, we select at most two gene expression profiles from each platform in the same experiment. After the selection procedure, total 442 gene expression profiles of 40 different cell types from 54 different experiment with 23 different platform types were remained  method. Another method compares common pathway set between two comparing pathway expression profiles converted from gene expression profiles. To measure the similarity between gene or pathway expression profiles, we used Spearman rank correlation test. Spearman rank correlation coefficient between profile X and Y is given byi = xi-yi, i = 1, ..., n and xi, yi = rank of ith gene or pathway in each profile X and Y. Spearman rank correlation coefficient ranges from -1 to 1, where similarity is maximum at 1 and minimum at -1.where dTo evaluate the performance of similarity measuring methods, we conducted cell type classification using nearest neighbor classifier. For each of 40 query gene expression profiles, the similarities to all of 402 gene expression profiles in search database were calculated. Then the profile with highest Spearman rank correlation coefficient was predicted to have the same cell type to query profile. Predicted profile was considered as correct prediction if its cell type is the same as that of query sample. If there is no same cell type profile in search database for a query profile, the search for the given query profile is not counted in classification accuracy assessment. Accuracy of classification is calculated by number of correct predictions divided by number of predictions.Similar profile search from the profiles of different platform or different experiment is harder than search from the profiles of the same platform or the same experiment . To evalWe conducted cell type classification using two different similarity measuring methods and access the performances with overall, cross-platform and cross-experiment search databases.Barplot shown in figure We next calculated average similarity score of top scoring hit for correct and incorrect classification cases Table . AverageWe analyzed further to figure out the reason for low classification accuracy of cross-experiment search. More specifically, our question is why cross-experiment searches show lower classification accuracy than that of cross-platform searches even though the similarity scores for top hits are higher than that of cross-platform searches. To answer this question, we divided cross-experiment search database more specifically into cross-experiment with the same platform profiles and cross-experiment with different platform profiles and conducted cell type classification with those two search databases. Table Reduced analytical dimension of pathway expression profiles from gene expression profiles might also contribute improved classification accuracy by pathway based methods. Not all genes in gene expression profile are converted to pathway expression profile for the incompleteness of current pathway information ,17. In cWe first attempted to use pathway information for gene expression similarity measurement. As previously developed pathway based gene expression analysis methods were successfully improve intact gene expression based analysis methods -23, pathConsequently, expression variations of multiple genes are summarized by a robust pathway expression, which represents the activity of the functional unit rather than a component of the unit, thus the pathway based methods result with higher classification accuracy, which demonstrates again that pathway level expression is more robust than individual gene level expression and pathway based similarity scoring methods could be successfully improve similar gene expression profile search.We demonstrated that our new gene expression similarity measuring method improved the precision of similar gene expression profile search when it's applied to cell type classification. We showed pathway expression profiling based similarity measuring method outperformed conventional gene expression profile based similarity measuring method over at most 50% for cross-platform profile search and 10% for cross-experiment profile search. At the same time, the classification accuracy shows that the methods still need to be improved, especially for searching similar profiles across different experiment. We believe that our research shed new light on similar gene expression profile search over rapidly growing large microarray databases by showing that integrating gene expression profile with external data such as pathway could improve search accuracy.CGSEP: Common Gene Set Expression Profile; PEPA: Pathway Expression Profile for All Gene set; PEPC: Pathway Expression Profile for Common gene set.The authors declare that they have no competing interests.CK designed whole research process, implemented all required methods for the research, analyzed results, drafted and revised the manuscript. JHW devised similarity scoring algorithm with CK and revised the manuscript. WSO and KTN helped the conceptualization of the research process. All authors read and approved the final manuscript.BMC Bioinformatics Volume 10 Supplement 15, 2009: Eighth International Conference on Bioinformatics (InCoB2009): Bioinformatics, available online at http://www.biomedcentral.com/1471-2105/10?issue=S15.Other papers from the meeting have been published as part of Dataset detail for cell type classification. All 442 gene expression profiles used for cell type classification are listed with detailed information. Each gene expression profile is annotated with three original GEO accession id, sample id(GSM), experiment id(GSE) and platform id(GPL), and cell type.Click here for file


In the process of tumour progression genetic instability is the basis for the evolution of tumour cell clones with various genotypic and phenotypic characteristics causing heterogeneity. Renal cell carcinoma has a long prediagnostic growth period, which increases the probability of clonal evolution. We have studied 200 consecutive renal cell carcinomas, addressing the interrelationship between intratumour heterogeneity and clinicopathological factors. DNA ploidy patterns were analysed in multiple samples from each tumour using flow cytometry and compared with clinical stage, tumour invasion, metastatic rate and survival. Eighty-five of 192 evaluable tumours (44%) were homogeneous concerning DNA ploidy . Among 107 heterogeneous tumours a majority (79%) contained aneuploid as well as diploid cell clones. Homogeneously diploid tumours had a lower incidence of local tumour spread compared with tumours with aneuploid cell clones (P < or = 0.001), but the frequency of distant metastasis at time of diagnosis was similar. The presence of aneuploidy in at least one sample from a tumour was a significant adverse prognostic factor (P < 0.001), whereas the degree of heterogeneity had no influence on survival. The frequent heterogeneity demonstrated indicates that multiple samples must be investigated to evaluate properly the malignant character of renal cell carcinoma.


A healthy 13-year-old boy who had spent the first 4.5 years of his life in an orphanage in Thailand before adoption by an American couple became ill with subacute sclerosing panencephalitis and died several months later. The boy had most likely contracted wild-type measles in Thailand. Measles complications are a risk in international adoptions.


Daniorerio.The vertebrate genome contains a predicted 50 000 100 000 genes, many of unknownfunction. The recent development of morpholino-based gene knock-down technologyin zebrafish has opened the door to the genome-wide assignment of function basedon sequence in a model vertebrate. This review describes technical aspects ofmorpholino use for functional genomics applications, including the potential formultigene targeting and known methodological limitations. The result ofsuccessful gene inactivation by this agent is proposed to yield embryos with a‘morphant’ phenotypic designation. The establishment of a morphant databaseopens the door to true functional genomics using the vertebrate,


PAF mediated lymphatic constriction was not altered by the intra-arterial infusion of phentolamine but was blocked by the intra-arterial infusion of the PAF receptor antagonist WEB 2170. These data indicate that in addition to PAF's effects on microvascular permeability, this agent may also impact the ability of the lymphatics to transport fluid through alterations in lymphatic smooth muscle tone. PAF mediated lymphatic constriction is not mediated by α-receptors but rather through PAF receptor mediated mechanism.Platelet activating factor (PAF) is a potent inflammatory lipid. In this study we assessed the ability of PAF to impact lymphatic vessel function by altering prenodal lymphatic resistance. Intralymphatic PAF (7.47 × 10


The authors report on 182 cases of intrathoracic metastases  observed as first single recurrences in the course of the follow-up of patients treated for primary breast cancer. ITM were detected on standard two-views chest X-ray (CXR) at regular follow-up visits and in absence of subjective symptoms (102 A cases) or in the interval between two consecutive planned controls because of the onset of subjective symptoms (80 S cases). The average disease-free interval since primary treatment was significantly shorter in A with respect to S cases  as a consequence of the early detection achieved by CXR survey. On the contrary, prognosis was not influenced by ITM early diagnosis as the 10-year survival since primary treatment did not differ significantly between A or S cases . Results were confirmed on multivariate (Cox's) analysis, adjusting for potential confounders such as age or nodal status. Periodic CXR survey looks a very questionable policy as it does not seem to have any favourable impact on prognosis. Its routine use in breast cancer patients should thus be carefully reconsidered.


The fish medaka is the first vertebrate capable of full spermatogenesis in vitro from self-renewing spermatogonial stem cells to motile test-tube sperm. Precise staging and molecular dissection of this process has been hampered by the lack of suitable molecular markers.We have generated a normalized medaka testis cDNA library and obtained 7040 high quality sequences representing 3641 unique gene clusters. Among these, 1197 unique clusters are homologous to known genes, and 2444 appear to be novel genes. Ontology analysis shows that the 1197 gene products are implicated in diverse molecular and cellular processes. These genes include markers for all major types of testicular somatic and germ cells. Furthermore, markers were identified for major spermatogenic stages ranging from spermatogonial stem cell self-renewal to meiosis entry, progression and completion. Intriguingly, the medaka testis expresses at least 13 homologs of the 33 mouse X-chromosomal genes that are enriched in the testis. More importantly, we show that key components of several signaling pathways known to be important for testicular function in mammals are well represented in the medaka testicular EST collection.Medaka exhibits a considerable similarity in testicular gene expression to mammals. The medaka testicular EST collection we obtained has wide range coverage and will not only consolidate our knowledge on the comparative analysis of known genes' functions in the testis but also provide a rich resource to dissect molecular events and mechanism of spermatogenesis in vivo and in vitro in medaka as an excellent vertebrate model. The testis is the male gonad where spermatogenesis takes place throughout adult life to continuously supply sperm for the next generation. Defects in testicular structure and function lead to testicular tumors and male infertility. In mammals, the adult testis consists of male germ cells and three major somatic cell types Much is known about the cell biology of spermatogenic germ cell development, which proceeds through three major stages: mitotic phase of proliferation and differentiation, meiosis and postmeiotic spermiogenesis This study aimed to establish a medaka testicular EST collection. For this, we generated a normalized cDNA library from the adult medaka testis and sequenced 7040 random EST clones. Comparative sequence analysis revealed a total of 3641 unique gene clusters.The expression levels of each individual genes in a genome can vary considerably in different cells, tissues or organs, and can even be very different in the same tissue or organ at different developmental stages or physiological conditions. The magnitude of difference can range from a few to as much as thousand folds. When RNA samples from an organism or an organ or a tissue are directly used for cDNA library construction, the differences among different genes in expression levels will normally be reflected in such libraries. If such a library is used for EST sequencing project, the problem of high redundancy will be brought in synaptonemal complex protein 3  gene and kallikreini gene (Leydig cell marker) was represented by six and two EST clones, respectively, in the 747 unnormalized EST collection whereas no corresponding clones was found in our 7040 EST clones. synaptonemal complex protein 1 gene  was represented by three EST clones in the 747 unnormalized ESTs and was represented by only one clone in our EST set. From the mathematic view, all these facts demonstrate the effectiveness of normalization in our library.A total of 8736 clones from the normalized cDNA library was sequenced, generating 7040 high quality (7033 having reads ≥200 bp read each) EST sequences (EMBO Accession numbers FM165707–FM172746). These 7040 sequences were subjected to clustering analysis using CLUSTAL W (1.83) Multiple Sequence Alignments program and 3641 clusters were obtained . Overallhttp://www.ncbi.nlm.nih.gov/UniGene/; dated at February 28th, 2008) ftp://ftp.ncbi.nih.gov/repository/UniGene/Oryzias_latipes/). The longest sequences in each of our EST clusters were retrieved and used to blast search against the NCBI medaka Unigene database, with the aim to assess the representation of our library in the public database. Unexpectedly, only a small fraction of our 3641 unique clusters  exhibit a match in the NCBI medaka UniGene database , and 2058 clusters were found to have their corresponding hits with high confidence in the non-redundant database   and S3.The 1197 genes (550 Unigenes and genes corresponding to 647 clusters) with known putative molecular function were combined and subjected to ontology analysis based on molecular and cellular functions. As expected, genes encoding for metabolic enzymes  form the major group in our EST set and in total 162 out of 1197 (∼14%) genes were recorded . A totalIt would be interesting to compare medaka testis-enriched genes with other vertebrates through genomic analysis. For example, are the genes conserved among the vertebrates? Is there any synteny relationship among these conserved genes? Are these genes alternatively spliced from annotated genes? Are there genes completely new without any annotated protein domains? However, these bioinformatic analyses rely on the availability of full length cDNA. This is mainly because cross-species comparison is normally performed on the amino acid sequence level but not at the nucleotide sequence level (because of the high variation of nucleotide sequences for ortholog genes between species during evolution). Use the amino acid sequences derived from full length cDNA will permit the identification of true conserved or novel genes. Then the in situ hybridization method can be used to prove if the novel gene is enriched in the testis. For this purpose, we designed a python program to retrieve information for the 1034 unigenes listed in Blimp1, Bmi1, YY1 etc) and receptors, ligands and signaling molecules  in testis , germ cell markers , pre-meiosis markers , meiosis regulator&structural protein , and postmeiosis marker (Msap and Rsh)   [29].pum1 (germ cell marker) and rad51 (meiosis marker), were examined via in situ hybridization in medaka testicular sections. The pum1 gene is expressed weakly in Spermatogonia at the periphery, but highly in primary spermotocytes and moderately in secondary spermatocytes  to the medaka EST collection and these new EST clusters/unigenes will be invaluable in assisting gene annotation once the medaka genome sequencing is completed. The wide range coverage of genes involved in diverse cellular activities and large number of novel sequences in our EST collection will not only consolidate our knowledge on known genes in testis st and 2nd stranded cDNA was synthesized using cDNA synthesis system . Oligo-(dT)20-V  was used as primer for the 1st strand cDNA synthesis. Two primers LLR1A (5′-gagatattagaattctactc) and LLR1B (complementary strand 5′-gagtagaattctaatat-3′) nd round PCR. Two more rounds of normalization were performed and the final PCR products were digested with EcoRI and ligated to pre-digested pBluescript SK+ vector for library construction. Colony picking and bacteria culturing are as described previously All procedures conducted with medaka fish are adhered to animal care guidelines  as outlined by the National Advisory Committee For Laboratory Animal Research in Singapore. Total RNA was extracted from pooled testes of adult medaka fish, using the Tri-Reagent according to the manufacturer's protocol . Total RNA was used for mRNA purification using mRNA purification kit . Both the 1Pasmid DNA was prepared in 96-well format using the conventional alkaline/SDS lysis method using robotics Biomek FX (Beckman) followed by ethanol precipitation http://www.mrc-lmb.cam.ac.uk/pubseq/manual/pregap4_unix_toc.html). In total 7040 ESTs (7033 with reads >200 bp) were obtained after editing (Table1) and clustering using Tigr-Assembler (http://www.tigr.org/software/assembler) identified a total of 3641 unique clusters  containing 17,307 unique clusters (released on February 2008). Sequences are considered identical if the blast E value is less than e−50ftp://ftp.ncbi.nlm.nih.gov/blast/db/nr). Only blast E value<e−8 were considered significant .The longest EST sequence in each of the 3641 unique clusters were retrieved and used as queries for BLASTN searches against the section Medaka UniGene Click here for additional data file.Table S2List of 1034 unique medaka testis EST clusters having hits in the medaka unigene database.(0.26 MB XLS)Click here for additional data file.Table S3List of 2057 unique medaka testis EST clusters having hit in the NCBI non-redundant database.(0.42 MB XLS)Click here for additional data file.Table S4Summary of ontology analysis of 1197 genes with known molecular/biochemical putative function(0.21 MB XLS)Click here for additional data file.Table S5List of 48 unigenes which have mRNA sequences in the unigene database.(0.02 MB XLS)Click here for additional data file.


Nuclear expression of TS in human tissue in vivo has not been characterised and its clinicopathological correlates in malignancy are unknown. 52 cases of primary colorectal carcinoma (CRC) and 24 cases of matched metastatic carcinoma were studied immunohistochemically using the monoclonal antibody TS106. The degree of nuclear TS immunostaining correlated closely with levels of TS mRNA expression amongst 10 CRCs studied. Strong nuclear immunostaining was seen in normal basal crypt colonocytes and germinal centre cells, and in a varying proportion of adenocarcinoma cells. Amongst the primary carcinomas, higher TS nuclear expression was associated with prominent extracellular mucin production and right-sided location. Higher TS nuclear expression also showed a significant association with poorer response to protracted venous infusional 5FU therapy. There was no clear association between TS nuclear expression and Ki67 or p53 expression assessed immunohistochemically. There was a strong positive correlation between TS nuclear expression in primary and metastatic CRC but the latter generally showed higher expression than matched primary tumour tissue. These findings confirm the nuclear expression of TS protein in human cells in vivo and provide new insight into how such expression may relate to the behaviour of CRCs. © 2001 Cancer Research Campaign


The cytotoxic and cytostatic responses of peripheral blood lymphocytes from eight cancer patients and splenocytes from four patients activated with rIL2 and a combination of rIL2 and rTNF-alpha were tested against two tumour cell lines. The cytotoxic response of rIL2-activated lymphocytes did not exceed the natural killer cytotoxicity values in all patients tested. In fact the killing capacity of some PBL deteriorated after rIL2 activation. The combined use of rIL2 and rTNF-alpha reversed this detrimental effect and enhanced the cytotoxic capacity of all PBL tested. In instances where high levels of killing were already achieved by rIL2 alone additional rTNF-alpha did not induce a significant change. This indicates that the role of rTNF-alpha may be to promote the response to rIL2 of PBL which react suboptimally to this lymphokine. rTNF-alpha did not only enhance cytotoxic capacity but also conferred cytostatic capacity to rIL2-activated LAK cells which were cytotoxic but unable to inhibit the growth of the surviving target cells. Natural killer cell selected K562 target cells which were less susceptible to killing by untreated lymphocytes than the parent K562 tumour cell line were killed more aggressively by rIL2 + rTNF-alpha LAK cells than by rIL2-LAK cells. No phenotypic differences were detected in these two cultures of LAK cells which indicates that the increased cytotoxic and cytostatic capacity of rIL2 + rTNF-alpha-LAK cells may be due to a higher state of activation of these cells or due to their capacity to recognise a broader spectrum of targets than rIL2-LAK cells.


Pericarditis as a differential diagnosis of the "Brugada-type Electrocardiographic Pattern Induced by Electrocution" reported by us   raised bHowever the early repolarisation pattern and non specific ST-T changes as a part of transient electrical injury of myocardium would be a better explanation for the ECG changes noted. The ST-T changes in right sided chest leads obviously simulates Brugada type pattern.The electrocution can result in non-specific electrocardiographic changes and varied ST-T changes mimicking pericarditis, early repolarisation, Brugada type pattern, myocardial infarction etc. However these changes needs to be correlated with the clinical background before the final diagnosis is made since majority of these changes are due to electrical injury of heart.


A 44 year old man presented with recurrent symptomatic palpitations,  always appearing during physical stress each with a similiar extent and never during resting periods. He also had fatigue and moderate dyspnoea during the episodes. The past medical history of the patient did not reveal any significant diseases. Findings of the attending physician including 12 lead ECG and echocardiography did not show any abnormality. As there was no electrocardiographic documentation of the episode, a 12-lead Holter evaluation was done.When the patient returned one week later, he reported of a typical episode of palpitations. The corresponding extract of the Holter ECG is shown in In synopsis of the findings we saw the indication for an invasive electrophysiology study due to a documented paroxysmal narrow complex tachycardia with alternating R-R intervals coming along with symptomatic palpitations.An octapolar catheter was placed in the coronary sinus with distal and proximal pair of electrodes configured as CS 1-2 and CS 7-8 respectively. Dual atrioventricular conduction properties of the AV node were verified with programmed atrial stimulation with premature coupling of one extrastimulus (« 500 S3 ») showing a typical AH-jump. No retrograde ventriculoatrial conduction was observed in our patient.Under basal conditions no tachycardia was inducable, so we decided to use an orciprenaline infusion to simulate physical strain as the tachycardia of our patient always appeared under these conditions. Finally, during orciprenaline infusion and atrial pacing via CS 7-8 with premature coupling of one extrastimulus (« 400 S3 »), a narrow complex tachycardia was induced  is a rare manifestation of dual atrioventricular nodal pathways. Two electrophysiological characteristics are basic requirements for a double ventricular depolarisation due to a dual AV node and simultaneous absence of the inducibility of a typical AV-nodal reentry tachycardia: [In conclusion, we have induced the clinical tachycardia of our patient. The patient underwent successful slow pathway modification with complete disapperance of symptoms and electrocardiographic manifestations of 1 : 2 atrioventricular conduction.


Clostridium difficile toxin alters colonic tissue levels of vasoactive intestinal peptide (VIP) at the expense of changes in colonic motility in the isolated perfused rabbit left colon. Colonic inflammation was induced by the intracolonic administration of 10−8 M C. difflcile toxin. Strain gauge transducers were sewn onto the serosal surface of the colon to evaluate colonic motility. C. difflcile administration produced histologic changes consistent with epithelial damage. This was associated with an increased production of prostaglandin E2 and thromboxane B2. Tissue levels of VIP but not substance P were significantly reduced. This was associated with an increased number of contractions per minute and an average force of each colonic contraction. These results suggest that tissue levels of VIP are suppressed by C. difflcile and may participate in colonic dysmotility during active inflammation.We investigated whether


Normal monkey tissues were found to contain an antigen which crossreacts immunologically with the carcinoembryonic antigen (CEA) of the human digestive tract. The monkey antigen reacted with complete or partial identity to the normal crossreacting antigen (NCA) in humans when tested in immunodiffusion against anti-CEA or anti-NCA. Extracts of monkey tissues inhibited in radioimmunoassays measuring human NCA. It is possible that monkey foetuses and colonic tumours contain CEA.


Forty-six patients who were treated with cisplatin or carboplatin for ovarian cancer developed resistant disease (no change in measurable disease or progressive disease) and 'crossed over' to the other platinum compound. Three patients (6.5%) responded to this second treatment but these patients had no survival advantage compared to the non-responders. One responder had progressive disease on cisplatin before crossing over to carboplatin.


A multistate outbreak of Escherichia coli O157:H7 infections occurred in the United States in June and July 1997. Two concurrent outbreaks were investigated through independent case-control studies in Michigan and Virginia and by subtyping isolates with pulsed-field gel electrophoresis (PFGE). Isolates from 85 persons were indistinguishable by PFGE. Alfalfa sprouts were the only exposure associated with E. coli O157:H7 infection in both Michigan and Virginia. Seeds used for sprouting were traced back to one common lot harvested in Idaho. New subtyping tools such as PFGE used in this investigation are essential to link isolated infections to a single outbreak.


A randomized trial comparing routine follow-up with a treatment regimen aimed at increasing specific anti-tumour immunity has been carried out in 95 patients after total surgical excision of lung cancer . Treatment consisted of inoculation with an autologous irradiated suspension of tumour cells combined with a small dose of C. parvum given intradermally during convalescence. Although treatment was associated with virtually no side effects, there has been no apparent benefit and input to the trial has now stopped.


Sir,Toxoplasma gondii (an intracellular coccidian parasite) is the most common cause of protozoon infections in humans and is highly prevalent in the adult population in the world and demonstrates various clinical manifestations throughout the world.[Mycobacterium tuberculosis antigen were negative. His chest X-ray and electrocardiography were normal. A repeat contrast MRI after 9 months showed the persistence of all the lesions and the patient was referred for a biopsy of the lesions. The patient underwent right frontal craniotomy and excision of the right frontal lesion. During surgery, there was well-defined firm, thick-walled lesion containing thick yellowish material. Histopathological examination of the specimen showed thick wall abscess and necrotic tissue with inflammatory cells in the center, features suggestive of toxoplasma cerebral abscess [3541111he world.–3 A 25-yhe world.. All the abscess . Further abscess 34 Severebscess 35 occasionbscess 358 The mosbscess 359 Althougbscess 35 As in thbscess 35 On imagibscess 3511 Howevescess 354 As in thess 35411 However,s 3541111 Spectross 3541111 Presumpts 3541111–16 As ins 3541111 The treas 3541111


The present study characterised p53 mutations in 44 hepatocellular carcinomas (HCCs) from Chinese patients residing in a high-incidence area. Twelve point mutations (27%) were detected in tumour tissues using single-strand conformation polymorphism analysis followed by direct DNA sequencing. Remarkably, no mutations were observed at codon 249. This is in contrast to HCCs from other high HCC incidence areas with endemic aflatoxin exposures, in which codon 249 is a mutational hot spot. It is therefore suggested that risk factors other than dietary exposure to aflatoxin may contribute to the high HCC incidence in Singapore.


Soft tissue sarcomas comprise a heterogeneous group of aggressive tumors that have a relatively poor prognosis. Althoughconventional therapeutic regimens can effectively cytoreduce the overall tumor mass, they fail to consistently achieve acurative outcome. Alternative gene-based approaches that counteract the underlying neoplastic process by eliminatingthe clonal aberrations that potentiate malignant behavior have been proposed. As compared to the accumulation of genealterations associated with epithelial carcinomas, sarcomas are frequently characterized by the unique presence of a singlechromosomal translocation in each histological subtype. Similar to the Philadelphia chromosome associated with CML,these clonal abnormalities result in the fusion of two independent unrelated genes to generate a unique chimeric proteinthat displays aberrant activity believed to initiate cellular transformation. Secondary gene mutations may provide an additionalgrowth advantage that further contributes to malignant progression. The recent clinical success of the tyrosine kinaseinhibitor, STI571, suggests that therapeutic approaches specifically directed against essential survival factors in sarcoma cellsmay be effective. This review summarizes published approaches targeting a specific molecular mechanism associated withsarcomagenesis. The strategy and significance of published translational studies in six distinct areas are presented. Theseinclude: (1) the disruption of chimeric transcription factor activity; (2) inhibition of growth stimulatory post-translationalmodifications; (3) restoration of tumor suppressor function; (4) interference with angiogenesis; (5) induction of apoptoticpathways; and (6) introduction of toxic gene products. The potential for improving outcomes in sarcoma patients and theconceptual obstacles to be overcome are discussed.


Hereditary non-polyposis colorectal cancer (HNPCC) is linked to an inherited defect in the DNA mismatch repair system. DNA from HNPCC tumours shows microsatellite instability (MSI). It has been reported that HNPCC patients have a better prognosis than patients with sporadic colorectal cancer. We examined whether the presence of MSI in a series of unselected colorectal tumours carries prognostic information. In a series of 181 unselected colorectal tumours, 22 tumours (12%) showed MSI. Survival analysis at 5–10 years follow-up showed no statistically significant difference in prognosis between MSI-positive and -negative tumours. Our results suggest that the MSI phenotype as such is not an independent prognostic factor. © 1999 Cancer Research Campaign


The incidence of nodal metastasis in early gastric carcinoma (EGC) is 10-20%. However, the optimal nodal dissection for early gastric carcinoma has not been established. A retrospective study was conducted in 392 consecutive patients who underwent potentially curative distal gastrectomy for EGC between 1962 and 1990. Of these 295 patients treated after September 1972 were prospectively entered into an extensive lymphadenectomy protocol. These patients were compared with 97 patients with simple gastrectomy in respect of the causes of death after surgery and the 10 year disease-specific survival rate. The incidence of nodal metastasis in early gastric carcinoma patients was 13.0%. Operative mortality from extensive lymphadenectomy was almost the same as from simple gastrectomy (2.0% and 2.1% respectively). Extensive lymphadenectomy provided a significantly higher 10 year survival rate than limited lymph node dissection . Among patients with nodal metastasis, the survival rate following extensive lymphadenectomy was significantly higher than that after simple gastrectomy . Among patients without nodal metastasis, there was no difference between the two groups in the survival rate . Multivariate analysis using the Cox proportional hazards model disclosed two significant independent prognostic factors on disease-specific survival, the nodal involvement  and the extent of lymph node dissection . Extensive nodel dissection appears to prevent recurrence and to improve the cancer-specific survival in EGC patients with nodal metastasis.


We report a rare case of obstructive jaundice caused by a tuberculous abscess in the head of pancreas,mimicking carcinoma. The case was successfully treated by pylorus-preserving proximal pancreatoduodenectomyand antituberculous drugs.


The thymic stromal cells alone induced positiveselection of functional CD4-CD8+ cells whose kinetics and efficiency were enhanced bynominal peptide. Fibroblasts expressing the selecting MHC alone did not induce positiveselection; however, together with nonselecting stroma and nominal peptide, there was inefficientpositive. These results suggest multiple signaling in positive selection with selection events ableto occur on multiple-cell types. The ease with which this model can be manipulated shouldgreatly facilitate the resolution of the mechanisms of positive selection in normal andpathological states.Presented here is a cell-suspension model for positive selection using thymocytes from


A long-finned pilot whale with morbilliviral disease was stranded in New Jersey. An immunohistochemical stain demonstrated morbilliviral antigen. Reverse transcriptase-polymerase chain reaction for morbillivirus P and N genes was positive. Novel sequences most closely related to, but distinct from, those of dolphin and porpoise morbilliviruses suggest that this virus may represent a third member of the cetacean morbillivirus group.


Acute inflammatory polyradiculoneuropathy secondary to chemotherapy for leukemia has been described in the pediatric literature. However, the reports are rare and have been mainly from intrathecal methotrexate in pediatric acute lymphoblastic leukemia patients who developed demyelinating polyradiculoneuropathy.A case report is presented of an unfortunate 53 year old Hispanic woman with acute myelogenous leukemia who developed profound weakness with cranial nerve palsies following both intravenous and intrathecal chemotherapy.This is an interesting and unusual case of predominantly axonal ascending sensory motor polyradiculoneuropathy with cranial nerve involvement in an adult patient with acute myelogenous leukemia following intravenous Cytosine arabinoside and intrathecal methotrexate. Polyradiculoneuropathies and ascending motor paraplegia have been described in pediatric leukemia patients undergoing chemotherapy. These rare reports have focused mainly on demyelinating polyradiculoneuropathies and/or myelopathies secondary to intrathecal methotrexate in children with acute lymphoblastic leukemia (ALL) . A case The patient was a 53 year old Hispanic woman who originally presented at the end May in 2007 with complaints of shoulder tightness and weakness in the neck and shoulders for approximately 3 weeks duration along with shortness of breath. She presented to a local community hospital where examination revealed an ill-appearing pale woman with tachypnia. Subsequent investigations showed thrombocytopenia with prominent leukocytosis and numerous blasts on peripheral smear plus disseminated intravascular coagulation with subsequent diagnosis of acute myelogenous leukemia  with hypoactive reflexes (0/4). Extra ocular muscles were intact without dysconjugate gaze, and the pupils were briskly reactive and symmetric.A lumbar puncture did not reveal any increased protein  or abnormal cells , however there were oligoclonal bands  and an elevated myelin basic protein . Cerebrospinal fluid cytology was negative for any malignant cells. An electromyogram and nerve conduction study showed absent sensory nerve action potentials and absent compound motor action potentials with diffuse denervation in all four extremities.The presumptive diagnosis was Guillain-Barré syndrome (GBS), and intravenous immunoglobulin (IVIG) was considered but felt to be contraindicated secondary to her newly diagnosed AML and stroke. She was therefore given 4 sessions of plasmapharesis which was held on the fifth session because of blood pressure fluctuations.Over the next ten days or so she did not improve neurologically, and in fact went on to develop right ptosis with a dysreactive pupil and outward eye deviation thought to be a third cranial nerve (CN 3) palsy, and a few days later bilateral facial weakness. A repeat lumbar puncture (about 2 weeks after the last one) again did not reveal any increased protein  or abnormal white blood cells (white blood cells were 0/cmm and red blood cells were 1/cmm) and the cerebrospinal fluid cytology was negative for any malignant cells.A sensory-motor neuropathy panel, myasthenia gravis panel, and a Lambert-Eaton myasthenic panel both were negative for any abnormal antibodies (Quest Diagnostics).A combined muscle and nerve biopsy was performed. The sural nerve showed peripheral neuropathy with prominent evidence of axonal degeneration with digestion chambers and secondary dropout of myelinated fibers which was moderately severe  ranges from 0.6 to 2.4 cases per 100,000 population per year .Although the most common neurotoxic effects of intravenous ARA-C involve the central nervous system, the association of ARA-C and polyneuropathy has also been reported with both intravenous ,5 and inPredominantly sensory peripheral neuropathy is seen with low dose ARA-C, whereas sensorimotor neuropathy occurs after high-dose therapy. Symptoms can start from hours to 2–3 weeks after initiation of therapy ,9-11 andAlthough central nervous system involvement is the most common neurotoxicity associated with intrathecal methotrexate therapy ,14,15, tWerner  reportedIt should also be mentioned that GBS can be associated with leukemia alone. Vembu and colleagues  reportedThe patient presented developed a third cranial nerve palsy which then progressed to a bilateral facial nerve palsy. At first it was felt that this may be a vasculitis with multifocal involvement. However, given the setting of her motor quadriplegia, respiratory failure along with the vasculitis-negative sural nerve biopsy results it was most likely a manifestation of AIDP or a GBS-like syndrome. Facial paresis, usually bilateral, is found in at least one half of patients  and otheAlthough there have been occasional pediatric case reports, this is an interesting case of ascending sensory motor polyradiculoneuropathy with cranial nerve involvement in an adult patient with AML following intravenous ARA-C and intrathecal methotrexate. The polyradiculoneuropathy appears to have been secondary to predominant axonal loss rather than demyelination as reported in most other studies.The author declares that he has no competing interests.RAR wrote the entire manuscript.Written informed consent was obtained from the patients' husband for publication of this case report and accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal.


We observed growing tumour vessels in vivo to study the relationship between blood flow and vascular enlargement. Mammary adenocarcinoma was implanted into Fisher-344 rat with dorsal skin-fold transparent chambers. Vascular growth was observed and recorded on videotape through a microscope for 6 h. Vascular networks were photographed and traced every 30 min to identify changes over time. Tumour sections were stained with Masson's trichrome and anti-Factor VIII-related antigen. Tumour growth was rapid enough for differences to be seen each hour. Vessels with a high blood flow showed an increase in diameter within a few hours and new branches formed from these vessels. In contrast, vessels without an increase in blood flow showed no change in diameter. Vessels within the interstitium surrounding the tumour were lined by endothelium that was positive for anti-Factor VIII-related antigen staining. Vessels in the tumour had extremely rare endothelial cells detectable by Masson's trichrome or anti-Factor VIII-related antigen staining. In conclusion, increased blood flow may cause vascular enlargement and some primitive vessels seem to lack endothelium.


Plasmodium falciparum involved sequestration of parasitized red blood cells and immunopathological responses. Among immune factors, IgG autoantibodies to brain antigens are increased in P. falciparum infected patients and correlate with disease severity in African children. Nevertheless, their role in the pathophysiology of cerebral malaria (CM) is not fully defined. We extended our analysis to an Indian population with genetic backgrounds and endemic and environmental status different from Africa to determine if these autoantibodies could be either a biomarker or a risk factor of developing CM.The main processes in the pathogenesis of cerebral malaria caused by P. falciparum infected patients manifesting mild malaria (MM), severe non-cerebral malaria (SM), or CM and in control subjects from Gondia, a malaria epidemic site in central India using quantitative immunoprinting and multivariate statistical analyses. We identified beta tubulin III (TBB3) as a novel discriminant brain antigen in the prevalence of CM. In addition, circulating IgG from CM patients highly react with recombinant TBB3. Overall, correspondence analyses based on singular value decomposition show a strong correlation between IgG anti-TBB3 and elevated concentration of cluster-II cytokine  previously demonstrated to be a predictor of CM in the same population.We investigated the significance of these self-reactive antibodies in clinically well-defined groups of P. falciparum infection and plasma cytokine patterns with clinical outcome of malaria. They also provide significant insight into the immune mechanisms associated to CM by the identification of TBB3 as a new disease-specific marker and potential therapeutic target.Collectively, these findings validate the relationship between antibody response to brain induced by


We report a patient who underwent radiofrequency ablation of the distal insertion of an atrio-fascicular accessory pathway with decremental properties because of inability to map a suitable potential alongside the tricuspid annulus. Small, discrete potentials resembling those of Purkinje fiber were found at right ventricular apex. All these potentials showed early activation during tachycardia preceding the QRS onset of various degree. Pace mapping helped to localize the presumed main distal insertion of the atrio-fascicular accessory pathway in a region where damage of the His-purkinje system may ensue. This case report describes catheter ablation of an atriofascicular accessory pathway by targeting its distal (ventricular) insertion site. Atrio-fascicular accessory pathways (AP) with decremental properties (i.e. Mahaim fibers) are associated with uncommon types of wide QRS complex tachycardia -5. In maA 44 years old male with structurally normal heart was referred to our institution because of long standing episodes of palpitation. Despite a history of sustained episodes of palpitation no evidence of arrhythmia was previously documented. The standard 12 leads EKG showed no baseline abnormality. The presence of structural heart disease was excluded by physical examination and trans thoracic echocardiography.After written informed consent was obtained an invasive EP study was performed in un-sedated, drug free state. One quadripolar and two decapolar diagnostic catheters were inserted through the right femoral and left subclavian veins and positioned respectively in the his bundle (HB) region, coronary sinus (CS) and  along the lateral wall of the right atrium. During the study the decapolar catheter located in the right atrium was moved to the right ventricular apex in order to record right bundle branch (RBB) potentials. During incremental atrial pacing, a progressive lengthening of the atrio-ventricular (A-V) interval was noted as well as progressive widening of the QRS and proportional shortening of the His-ventricular (H-V) interval.  A wide complex tachycardia exhibiting left bundle branch block (LBBB) morphology and superior axis was induced during pacing at 240 milliseconds (msec) from the high right atrium. The appearance of LBBB morphology was invariably associated with the progressive shortening of the HV interval up to the merging of the His potential with the local ventricular electrogram. After a stable RBB potential recording was obtained it was shown an inversion of the physiologic HB–RBB activation sequence during tachycardia, the HB resulting activated retrogradely. This tachycardia was characterized by an A:V ratio of 1:1 and a concentric retrograde atrial activation pattern . Programmed atrial stimulation at different drive cycles (600-400 msec.) with double and triple extrastimuli successfully initiated tachycardia at coupling interval of less than 300 msec. The arrhythmia onset was always preceded by the occurrence of LBBB aberration of the last paced beat. During the recording session tachycardia cycle length (TCL) varied from 290 msec to 340 msec. These TCL variations were associated to proportional prolongations of V-A interval and QRS duration while the atrial retrograde activation sequence remained unmodified. Despite TCL lengthening and VA prolongation the HA interval remained constant. Atrial pacing at cycle length shorter than the TCL up to 240 msec successfully entrained the tachycardia without obtaining sinus rhythm restoration after pacing interruption. Rapid atrial pacing during sinus rhythm also initiated a self-limiting typical counterclockwise atrial flutter with 2:1 A-V pre-excited response. Ventricular pacing from the parahissian region at a cycle length just shorter than TCL invariably interrupted the tachycardia after His bundle capture was obtained. The retrograde atrial activation in the His bundle trace always preceded the others and the atrial activation sequence remained unmodified during the entire recording session even in the presence of  TCL changes, this behavior was consistent with true antidromic AV reciprocating tachycardia using the Mahaim fascicle as antegrade limb. V-A prolongation without H-A modification excluded the involvement of a septal concealed accessory pathway as retrograde limb and also ruled out an atrioventriclar nodal reentrant tachycardia with a Mahaim fascicle participation as an innocent "bystander". We may speculate that this behavior probably reflected paroxysmal prolongation of the conduction time or block thorough the RBB next to the insertion of the Mahaim fiber.The ablation procedure was performed using a 7 F 4 mm ablation catheter . A careful and complete tricuspid ring mapping was performed in order to record a discrete potential resembling that recorded on His region, the so called Mahaim potential (MP). We failed to identify a discrete potential alongside the entire annulus so we decided to target the ventricular insertion. Given the superior axis of the arrhythmia we deliberately started from the right ventricular apex. Small, discrete potentials resembling those of Purkinje fiber were found near the right ventricular apex, these potentials showed various degrees of early activation during tachycardia and all preceded the His bundle activation. We delivered RF at a site with the apparently most negative local potential-QRS interval but we failed to terminate the arrhythmia. In order to reduce the number of unnecessary RF pulses we decided to perform pace mapping in a region where different sites with early local ventricular activation could be recorded. Pace mapping performed from the recording site of a small potential (SP), presumably the distal RBB, located 35 msec before the tachycardia QRS onset and 40 msec before the HB retrograde activation showed a QRS morphology identical to that observed during tachycardia. ,3.During sinus rhythm, pacing from this site was also followed by tachycardia initiation and of note the stim-QRS interval resembled the SP-QRS interval during tachycardia. This behavior was consistent with the antegrade limb distal insertion of the macroreentrant circuit. A single pulse of RF  interrupted the tachycardia after less than 3 sec, the RF delivery was then continued for 60 seconds in order to achieve a more complete lesion. At the end of the procedure the arrhythmia was no longer inducible, incremental atrial pacing demonstrated normal AV conduction and an incomplete RBB block was evident. Patient was followed up for one year and there were no recurrences of tachycardia.Recently unsuccessfull tricuspid annulus mapping for Mahaim discrete potential has been reported to be up to nearly 50 % . When th


Mice have been compeletly protected against the lethal effects of repeated injections of methotrexate by 5-methyltetrahydrofolic acid. Methotrexate-resistant cells probably owe their resistance to their high content of this substance.2. Daily small doses were much more effective than fewer but larger doses.3. The reduced efficiency found in counteracting high dosages of methotrexate may be due to interference with folate transport across cell walls.4. 5-Methyltetrahydrofolic acid and citrovorum factor did not differ greatly in their ability to inhibit methotre toxicity. It is suggested that the pharmaceutical effects of citrovorum factor are due to its conversion to 5-methyltetrahydrofolic acid.


This article describes the chemical aminoacylation of the yeast phenylalanine suppressor tRNA with a series of amino acids bearing fluorinated side chains via the hybrid dinucleotide pdCpA and ligation to the corresponding truncated tRNA species. Aminoacyl-tRNAs can be used to synthesize biologically relevant proteins which contain fluorinated amino acids at specific sites by means of a cell-free translation system. Such engineered proteins are expected to contribute to our understanding of discrete fluorines’ interaction with canonical amino acids in a native protein environment and to enable the design of fluorinated proteins with arbitrary desired properties. Over the past two decades, the interest in engineering proteins containing site-specific synthetic amino acids with novel functionalities has grown considerably. The utility of chemically aminoacylated suppressor transfer RNAs (tRNAs) combined with cell-free translation systems in producing proteins that contain non-canonical amino acids was reported independently from each other by Schultz and Chamberlin –2. TheirNα-protected amino acids were used to chemically aminoacylate the dinucleotide pCpA. Subsequent enzymatic ligation to truncated tRNAs  yielded the desired AA-tRNAs . Th. Thn-butThe efficient chemical and enzymatic synthesis of three novel fluorinated aminoacyl-pdCpAs and Abu-pdCpA and their corresponding charged tRNAs is reported. These aminoacyl-tRNAs can be used for site-specific protein mutagenesis in a cell-free protein synthesis system and will enable a systematic investigation of the structural and dynamic behavior of fluorine within a native protein environment.Supporting information features detailed information on experimental procedures and compound characterization.File 1Experimental procedures and compound characterization


Fifty-three biopsies from the oral mucosa in 51 Indian villagers with oral submucous fibrosis have been studied for epithelial changes. A marked atrophy of the epithelium was found in 71.7%; the atrophic epithelium showed no rete ridges. The normally unkeratinized buccal mucosa revealed either ortho- or parakeratosis or hyperortho- or hyperparakeratosis in 48.0%. In one patient the histologic examination disclosed a squamous cell carcinoma. In 22.6% an epithelial atypia was found. The histologic features in these atypias are compared with those seen in leukoplakias from the same survey. The possible precancerous nature of submucous fibrosis is discussed.


Vesicoureteric complications present early after transplantation and contribute to patient morbidity, graft loss and mortality. Ureteral stenting provides a decrease in ureteroneocystostomy anastomotic complications following renal transplantation. There should be prophylactic stent insertion with endoscopic removal at a designated time post transplantation. With the addition of antibiotic prophylaxis post transplantation, ureteric stenting does not increase the rate of urinary tact infections. There is no significant increase in cost for stenting during transplantation in comparison to management of major ureteric complications. Routine stenting causes significant cost-saving per year and prevents anastomotic complications. It is wise to stent the transplant ureter routinely. Kidney transplantation is the treatment of choice for end-stage kidney disease. Ureteroneocystostomy anastomotic leakage and/or strictures complicate 3–9% of all renal transplants.–3 These We would like to address the following three categories:Initially, ureterovesical anastomosis was done using a transvesical Leadbetter-Politano approach which is presently superceded by the extravesical ureteroneocystostomy (Lich-Gregoir) approach.–16 RefinConventional native ureteric repairs over stents are widely accepted to have a better outcome. In addit26In all randomized and quasi-randomized controlled trials looking at the use of double-J stents to prevent urological complications, the incidence of MUCs ranged between 0 and 5% in stented patients (median 1.0%) and between 0 and 17.3% (median 7.0 %) in the non-stented patients .5–11] A A11] A tThe most significant theoretical complication in the use of a stent is an increase in the number and severity of urinary tract infections (UTIs). Other possible complications include persistent hematuria, bladder discomfort, stent migration, breakage, encrustation and complications during removal. Most centers have adopted a policy of prophylactic stent insertion with endoscopic removal at a designated time post transplantation in an effort to reduce the rate of MUCs. UTIs, inThe maximum reported non-infectious complications were irritative symptoms 5.6%, breakageIn another study it has been suggested that ureteral stasis may cause tubuloepithelial injury and slow down the decrease in creatinine levels. They concluded that Double J Stent (DJS) did not increase UTIs but provided a smooth decline in creatinine levels, which may reduce the question of acute rejection.The incremental inpatient hospital costs associated with a urinary complication during the first 12 months following renal transplantation were 145% of the cost of renal transplantation without this complication. Notably, this value does not include inpatient hospital indirect costs, any expenses generated on an outpatient basis or elsewhere, or any inpatient or outpatient professional costs. Thus, the actual incremental cost associated with a urinary anastomotic complication following renal transplantation is significantly higher than this value. There isP <0.0001).[P = 0.02).[A recent meta-analysis evaluated five prospective, randomized, controlled clinical trials of routine stenting following renal transplantation and indicated that the collective urinary complication rate following routine stenting was 1.5% compared to 9% without stenting .The review of the literature appears to tilt the balance heavily in favor of routine prophylactic stenting in renal transplant recipients. Transplant units currently using antibiotic regime as prophylaxis for pneumocystis carnii should not notice an excess of stent-related infections. The use of an appropriate size of stent and early removal at two weeks prevents morbidity and stent-related complications. It is wise and cost-effective to stent the ureteroneocystostomy after transplantation.


Pulmonary edema after device closure of atrial septal defect (ASD) is a rare complication. We present illustrative images of a case of pulmonary edema after device closure of ASD in a 53 year old adult. Older patients undergoing ASD closure can benefit from their left atrial and left ventricular end diastolic pressures measurement before and after temporary balloon occlusion of ASD. These images illustrate the development of pulmonary edema after catheter closure of atrial septal defect (ASD) in a 53-year-old gentleman. He presented with shortness of breath and easy fatigability since six months. He was normotensive and non-diabetic with no history of effort angina. The chest X-ray obtained before the procedure  showed cEighteen hours after device closure the patient developed breathing difficulty and orthopnea. Clinical examination showed tachypnea and basal crepitation. Skiagram of chest in postero-anterior view showed features of evolving pulmonary edema  which eset al.[et al. reported pulmonary edema in a 78-year-old male. Our patient was relatively younger (53 years) without any coronary artery disease and normal systemic pressure.ASD is a common form of congenital heart disease accounting for approximately 10% of all congenital cardiac defects.[et al. reportedIn older patients, an impaired diastolic ventricular function due to reduced ventricular compliance is observed more frequently although this is occasionally also seen in childhood. RestrictOlder patients undergoing ASD closure in the catheterization laboratory should perhaps have their left atrial, LVEDP measured before and after temporary balloon occlusion and after defect is closed with a device.


Geminiviridae) are small single-stranded (ss) DNA viruses infecting plants. Their virion morphology is unique in the known viral world – two incomplete T = 1 icosahedra are joined together to form twinned particles. Geminiviruses utilize a rolling-circle mode to replicate their genomes. A limited sequence similarity between the three conserved motifs of the rolling-circle replication initiation proteins (RCR Reps) of geminiviruses and plasmids of Gram-positive bacteria allowed Koonin and Ilyina to propose that geminiviruses descend from bacterial replicons.Geminiviruses (family Satellite tobacco necrosis virus was found to be the best template for homology-based structural modeling of the geminiviral capsid protein. Good stereochemical quality of the generated models indicates that the geminiviral capsid protein shares the same structural fold, the viral jelly-roll, with the vast majority of icosahedral plant-infecting ssRNA viruses.Phylogenetic and clustering analyses of various RCR Reps suggest that Rep proteins of geminiviruses share a most recent common ancestor with Reps encoded on plasmids of phytoplasmas, parasitic wall-less bacteria replicating both in plant and insect cells and therefore occupying a common ecological niche with geminiviruses. Capsid protein of We propose a plasmid-to-virus transition scenario, where a phytoplasmal plasmid acquired a capsid-coding gene from a plant RNA virus to give rise to the ancestor of geminiviruses. The origin(s) of viruses is a longstanding but yet unresolved question in biology. Several hypotheses were put forward in efforts to understand this enigma (reviewed in ). AccordCircoviridae family bear only two genes , which has been previously observed to be similar to Reps of circoviruses . The pairwise distances between the nanoviral and other Reps were considerably larger than distances between any other pair of sequences (data not shown). Examination of the sequence alignment [see Additional file The 40 sequences were aligned. The alignment was manually verified and edited [see Additional file N. tabacum [B. catenulatum DSM 16992 on the ML tree was different from that on the Bayesian tree.Maximum-likelihood (ML) and Bayesian trees were inferred using PhyML  and MrBa tabacum ) form a YP_001586272] and pUB110 [GenBank:CAA27141], the two plasmids whose Reps were found to be the closest to geminiviral Reps , 69.1% had corresponding amino acids in at least one of the four geminiviral CP sequences   Fig. . Given tMicroviridae and Parvoviridae families  was previously shown to be more similar to Reps of various circoviruses than it is to Reps from other bacterial plasmids and viruses /[LM]T) [YN]XHX [HQ]XYXXKXGXXXXGK [ST]XDD. The residues shown in square brackets are alternatives; X – any amino acid; numbers in parentheses denote the allowed distance between corresponding motifs; slash sign indicates alternation of the dipeptides in the second and third positions in the pattern. The non-redundant protein sequences and environmental protein sequences from BLAST database were downloaded (07.02.2009) from NCBI FTP site and searched for sequences exactly matching the derived pattern without paying attention to the sequences surrounding the conserved motifs . Using this approach sequences missed by BLAST searches are expected to be found. 1072 protein sequences were initially extracted. In order to avoid redundancy, the original dataset was subsequently filtered to leave only sequences with less than 70% identity. As a result, a dataset containing 43 protein sequences was obtained. Of these two sequences were false-positive – a 799 amino acid-long hypothetical protein [GenBank:XP_001614627] from Plasmodium vivax SaI-1 and a 440 amino acid-long hypothetical TrmE domain protein GOS_1133298 [GenBank:EDE42344] from marine metagenome project, which were not included in the further analysis. The resultant dataset (41 sequences) was used to create a multiple sequence alignment using CLUSTALW [ABD67440] was found to be considerably longer (469 aa) than the rest of the sequences. The protein was found to be a fusion of RCR Rep and geminiviral transcriptional activator AC2 and was therefore removed from the alignment. The 40 sequences were realigned and following manual examination and editing the subsequent alignment [see Additional file 6 generations and trees were sampled every 100 generations. To construct the consensus tree, the first 25% of the trees were discarded as "burnin".Koonin and Ilyina 1992) found that geminiviral rolling-circle replication (RCR) initiation proteins (Rep) are related to certain bacterial Reps . In ordeCLUSTALW . One gem992 found100)YXXKX,200GXXXXMultiple sequence alignment [see Additional file BioInfoBank MetaServer  was usedMK conceived the project, collected, analyzed and interpreted the data, and drafted the manuscript. JJR collected and analyzed the data, and revised the manuscript. DHB interpreted the data and revised the manuscript. All authors read and approved the final manuscript.Multiple sequence alignment of 40 RCR Rep proteins. Figure shows a multiple sequence alignment which has been used to calculate the phylogenetic trees.Click here for fileBayesian consensus tree of the RCR Rep proteins. Figure shows the Bayesian consensus tree which has been calculated using the same dataset as for the Maximum likelihood tree shown in Figure Click here for file


At the moment only early detection of lung cancer offers a good prognosis for the patients. Conventional white light endoscopy is mostly insufficient for early diagnosis. Therefore wedeveloped a system of fluorescence diagnosis using 5-aminolaevulinic acid (ALA)exogeneously applied. As precursor of the heme synthesis it is metabolized to protoporphyrinIX –  a red fluorescent substance. Therefore protoporphyrin IX accumulates in tumorous andpremalignant tissue, and can be directly visualized by fluorescence bronchoscopy. Excitationwith blue light (380–435 nm) causes a red fluorescence, which can be detected after filteringmost of the blue component with the naked eye or a camera system. After earlier work withlaser systems and cold light sources we now use the system D-Light AF for the fluorescencediagnosis using ALA-induced protoporphyrin IX fluorescence.


Among gastric cancer patients, the Rhesus D-negative phenotype correlated with increased tumour recurrence  and reduced overall survival time  and was identified in multivariate analysis as the most important independent prognostic marker in the R0 patient group .


Biliary complications continue to be a major cause of morbidity after split-liver transplantation (SLT). In this report we describe an uncommon late biliary complication. One year after SLT the patient showed an intrahepatic bile dicy dilatation with severe cholangitis episodes. The segmentary bile duct of hepatic segment VI-VII draining in the left duct was unidentified and tied at the time of the in situ split-liver procedure. We perform a permanent obliteration of the dilated intrahepatic ducts by a percutaneous embolization using an n-butyl cyanoacrylate (NABC). The management of biliary complications after SLT requires a multidisciplinary approach. The use of NBCA in obliteration of a dilated bile duct seems to be a safe procedure with good results providing a less invasive option than hepatic resection and decreasing the morbidity associated with chronic external biliary drainage. Further studies are needed to determine whether this approach is effective and safe and whether it could reduce hospital stay and cost. Split-liver transplantation (SLT) is an attractive alternative procedure to expand the donor pool in patients waiting for liver transplantation. Paramount to the success of SLT is a careful donor and recipient selection. The standard split-liver procedure for a child and an adult , by splitting segment II-III for pediatric recipient and segment I-IV-V-VI-VII-VIII for an adult, is an accepted surgical option with good results both for the adult and for pediatric recipient .Biliary complications continue to be the major cause of morbidity after SLT with a reported high incidence ranging between 10% and 32% [As a consequence of bile duct anatomic variations, SLT requires a precise knowledge of the liver anatomy , 4. The In this report we describe an uncommon late biliary complication that occurred after SLT and was successfully treated by a multidisciplinary approach. A 63-year-old male with hepatitis C-related cirrhosis was referred for liver transplantation to our institution. We performed a conventional A/P SLT with in situ technique providing the left lateral segment for a child (segments II-III) and leaving the right lobe graft (segment I-IV-V-VI-VII-VIII) for an adult recipient. The celiac trunk was left on the left graft while the right hepatic artery remained on the right graft. The common hepatic bile duct was left on the right graft. The patient was transplanted using the piggy-back technique without a veno-venous bypass. The biliary tract was reconstructed performing a duct-to-duct anastomosis using a T-tube by our standard technique previously described .A cholangiography through the T-tube was performed on postoperative day 14, and the T-tube was clamped before patient discharge. Three months after A/PSLT the T-tube was removed after a cholangiography with normal findings. One year after transplant the patient showed abnormal liver function tests, hyperbilirubinemia, leukocytosis, and elevated g-glutamyl transpeptidase (GGT), and mild elevation in alanine transaminase (ALT). The patient underwent a doppler ultrasound that showed (a patent hepatic artery) an intrahepatic bile duct dilatation and an anastomotic biliary stricture. These findings were confirmed by a magnetic resonance cholangiography (MRC). The anastomotic stricture was treated by stenting the main biliary duct during an endoscopic retrograde cholangiopancreatoghaphy (ERCP) without any evidence of intrahepatic biliary dilatation. After this procedure the patient was submitted to a percutaneous transhepatic cholangiography (PTC) showing a complete obstruction of segments VI and VII biliary branches near the duct-to-duct biliary anastomosis . A percuWe supposed that the segmentary bile duct branch of posterior segments VI and VII draining in the left bile duct was unidentified and tied at the time of the in situ split-liver procedure during the parenchymal transection. A primary biliary reconstruction by biliary repair or biliodigestive anastomosis was considered at high risk because of the presence of postsurgical adhesions sand due to fibrotic tissue replacing the parenchymal transected area. A liver resection of the dilated segments VI and VII was considered at high risk of leaving an inadequate liver mass. We decided to perform a permanent intraoperative obliteration of the dilated intrahepatic ducts by a percutaneous embolization using a nonresorbable agent. With a fluoroscopic guidance through the transhepatic access we positioned a 5-French polyethylene catheter inside the ducts, preliminary flushed by a nonionic dextrose solution. We then injected the tissue adhesive agent n-butyl cyanoacrylate  mixed with ionized oil  for opacization in a ratio of 1:5. This solution completely filled the biliary duct, and the occlusion was totally accomplished in a few seconds .During the first 3 days after the chemical bile duct embolization, the patient had a low fever with a slightly abnormal liver function test. A computed tomography (CT) scan performed 6 months later showed no sign of hepatic abscesses, and the bile duct dilatation was completely occupied by the NBCA-Lipiodol mixture. One year after the procedure patient showed normal liver function tests without no episodes of cholangitis.The management of biliary complications after liver transplantation requires a multidisciplinary approach.Chemical bile duct embolization treatment could represent a valuable solution to treat uncommon biliary complications. These tissue adhesive glues are low-viscosity liquid monomers that undergo rapid polymerization and solidification when they come into contact with organic fluids such as bile. NBCA is a permanent liquid embolic material that produces long-term occlusion in vessels of various size through an inflammatory tissue response resulting in vessel thrombosis or tissue atrophy.Little is known about the use of cyanoacrylate compounds, and unlike European countries the use of Glubran has not been approved by the Food and Drug Administration yet. Other authors have described the efficacy of biliary duct ablation by NBCA in patients with persistent postsurgical bile leaks after lobectomy or cholecystectomy. Vu et al. treated six patients with persistent postsurgical bile leaks as a complication after hepatic lobectomy or cholecistectomy using NBCA glue for the obliteration of isolated segmental bile ducts in four cases . EndoscoThe percutaneous interventional technique represents an effective valuable approach to reduce mortality and morbidity in the treatment of biliary complications after liver transplantation.The use of NBCA in obliteration of a dilated bile duct seems to be a safe procedure with good results providing a less invasive option than hepatic resection above all in high-risk patients with posttransplant bile duct injuries, decreasing the morbidity associated with chronic external biliary drainage.Futher studies are needed to determine whether this approach is effective and safe and whether it could reduce hospital stay and costs.


The NKactivity of PBL was detected by a cytotoxic test using3H-uridine human erythromyeloblast K562 cells. AutologousIFN-α and IFN-γ did not augment NK activity of PBL fromhealthy donors in vitro, whereas in samples from MSpatients the IFNs strongly stimulated NK cell cytotoxic function.This stimulation suggests the existence of an inhibitor ofregulatory IFN action, that is produced in healthy donorssimultaneously with IFN in response to IFN induction, but which islacking in commercial IFN preparations. The factor-containingsupernatants from healthy donors reduced the stimulatory action ofautologous IFNs in patients with MS almost until complete blockade.Because this inhibitor was absent in patients with MS, deficiency ofan inhibitor of IFN regulatory action in MS could open the way totreatment of this compartment of the immune system.The


Sera from Balb/c mice bearing Moloney leukaemia block complement dependent antibody mediated cytotoxicity of an antiserum prepared in rats against syngeneic Moloney virus induced lymphomata when either spleen cells from mice bearing Moloney leukaemia (M) or an in vitro line of Moloney virus transformed cells (MSC) are used as targets. This antiserum has been shown to recognize p30, the major internal virion protein, as a cytotoxic target on these cells. Viral particles were identified by electron microscopic examination of pelleted material obtained from leukaemic sera after high speed centrifugation. However, removal of virus did not affect the capacity of the leukaemic sera to absorb cytotoxicity of rat ILR-3 for MSC targets, and only depressed somewhat its ability to absorb activity of the same antisera against M targets. Virus-free leukaemic sera also blocks complement dependent antibody mediated cytotoxicity of an antiserum prepared in goats against the gs3 determinant of p30. This indicates that the material in leukaemic sera responsible for the in vitro block of antibody mediated cytotoxicity was p30. A lesser degree of block was observed with sera obtained from normal Balb/c mice, but the nature of material responsible is as yet undefined.


When multiple synchronous lung tumours are identified, discrimination of multicentric lung cancers from intrapulmonary metastases by clinical findings is often difficult. We used genetic alterations in p53 gene as a discrimination marker of double primary lung cancers from single lung cancer with intrapulmonary metastasis. Twenty of 861 patients with primary lung cancer who underwent lung resection were selected as subjects because they showed synchronous double solid tumours of the same histological type in the unilateral lung without distant metastases. In addition, they had been diagnosed as lung carcinoma with intrapulmonary metastasis by clinical and histological findings. DNAs were extracted from paraffin-embedded tissue of paired tumours from these 20 patients. Exons 5–9 of the p53 gene were examined for genetic alterations in the tumours by polymerase chain reaction, single-strand conformation polymorphism analysis and subsequent DNA sequencing analysis. Three different patterns in the distribution of p53 mutations in double lung tumours were observed: [A] mutation in only one of the tumours (four cases), [B] different mutations in the tumours (two cases), and [C] same mutation in both tumours (one case). The cases of [A] or [B] patterns could be classified as double primary lung cancers, while the case of the [C] pattern was suggested to be lung cancer with intrapulmonary metastasis. These results suggested that the multicentric cancers were more frequent than the intrapulmonary metastatic cancers in double cancer cases. © 1999 Cancer Research Campaign


The Río Tinto (RT) is distinguished from other acid mine drainage systems by its natural and ancient origins. Microbial life from all three domains flourishes in this ecosystem, but bacteria dominate metabolic processes that perpetuate environmental extremes. While the patchy geochemistry of the RT likely influences the dynamics of bacterial populations, demonstrating which environmental variables shape microbial diversity and unveiling the mechanisms underlying observed patterns, remain major challenges in microbial ecology whose answers rely upon detailed assessments of community structures coupled with fine-scale measurements of physico-chemical parameters.By using high-throughput environmental tag sequencing we achieved saturation of richness estimators for the first time in the RT. We found that environmental factors dictate the distribution of the most abundant taxa in this system, but stochastic niche differentiation processes, such as mutation and dispersal, also contribute to observed diversity patterns.We predict that studies providing clues to the evolutionary and ecological processes underlying microbial distributions will reconcile the ongoing debate between the Baas Becking vs. Hubbell community assembly hypotheses. Geological and geochemical studies show the Río Tinto to be an acidic river situated at the core of the largest Pyritic Belt on Earth  whose chet al.Vibrio spp., these clusters constituted individuals different at the genomic level but whose divergence should be neutral (i.e. with no selective advantage) because of the small spatial scale in which they coexisted Clustering sequences into operational taxonomic units (OTUs) is the first step in a molecular study exploring ecological diversity. Microbiologists traditionally use a 97% similarity cut-off value to form OTUs that delineate microbial species Acidithiobacillus ferrooxidans and Leptospirillum ferrooxidans and other relatively less abundant ribotypes similarity among samples through the use of multivariate methods that compare samples based on taxon composition and relative abundance. It is important to note that taxonomic description is not required for assessment of either of these measures.In order to generate accurate estimates of diversity, our sampling scheme consisted of a replicated sampling design and large area of coverage of the stations observed microbial richness among samples at the same sampling effort overall species richness and compare alpha diversity of communities provided they reach an asymptote et al. have suggested a new set of statistical approaches to calculate microbial richness from parametric models Each method for estimating richness and comparing alpha diversity between samples has advantages and drawbacks To evaluate relationships among samples based on shared OTU relative abundance, we present results using the Morisita-Horn pairwise similarity coefficient. This index is widely used because it is less influenced by species richness and sample size than other (dis)similarity measures of quantitative data et al.A. ferrooxidans and L. ferrooxidans obtain their energy from pyrite (FeS2) and the oxidized metabolites can in turn be mineralized by heterotrophic microbes like Acidiphilium spp., Ferromicrobium or sulfate reducers. Ferric iron buffers the pH at or below pH 3. In our study, of the 22 environmental parameters measured, seven best explained the variation in the data  (see Amils dox) see . We useddox) see . Monte Cdox) see . OTUs thdox) see  and 5. Fdox) see . We infeet al.Stochastic vs. deterministic community assembly hypotheses are being tested in parallel to explain the distributional patterns of organisms in natural environments Candidatus captivus acidiprotistaeglobal dispersion of ecotypes better adapted to certain environmental conditions, which prevail over less adapted units that emerge locally. Pommier et al. (2007) and Ramette et al. (2007) have suggested a similar pattern of global deterministic ecotype adaptation On the other hand, the presence of endemic ecotypes that correlate with particular environmental factors seem to contradict Baas Becking's ideas on global bacterial distribution Seasonal sampling that integrates bacterial, archaeal and eukaryal components of the microbial community is the necessary next step to understanding whether interaction of all trophic levels in the RT confirm or reject our scenario of the global distribution of adapted ecotypes.Our study included three stations in the RT that have distinct physico-chemical parameters and biology We added Cell Lysis Solution from the Puregene® DNA extraction kit  directly to the sterivex filter using a 3cc. syringe, sealed the filter, and placed it into a liquid nitrogen dry shipper . We extracted total DNA within one week of collection using the Puregene Bacteria DNA purification procedure with the following modifications. We added lysozyme (67 µL of 50 mg/mL solution) and proteinase K (10 µL at 20 mg/mL) consecutively directly to the sterivex filters and incubated these enzymes with agitation as indicated in the protocol. We then transferred incubated samples to three 2-mL microfuge tubes to proceed with the protein precipitation step. Nucleic acid precipitation occurred in 1 volume of isopropanol. Finally we resuspended DNA pellets in 30 µL Puregene© DNA Hydration Solution per sample and stored them at −20°C until further processing.For each sample both filtered and unfiltered 15 mL water aliquots were analyzed using Total Reflection X-ray Fluorescence (TXRF) at the Universidad Autónoma de Madrid  to determine the concentration of 22 chemical elements in the water samples examined. Given filtered vs. unfiltered geochemistries were not significantly different, we report results with unfiltered samples.in situ in the river at the time of water collection. These two parameters varied with time of day as the sun rose and heated the river so they were not included in our analyses.We measured redox potential and pH (using a Crison 506 pH/Eh meter) and conductivity (using a Orion-122 conductivity-meter) at the time of water collection from 15 mL aliquots. Oxygen concentration and water temperature were measured using an Orion-810 oxymeter http://www.ncbi.nlm.nih.gov/GeneBank) and RDPQuery SARST-V6 produces sequences of large concatemers of PCR-amplified ribosomal sequence tags (RSTs) from homologous V6 hypervariable regions. We performed amplification and purification of the V6 region of bacteria following We first compared diversity between samples by representing relative abundances of OTUs in a rank/abundance plot. We then tested whether the data fit one of four statistical models of species abundance distributions: the geometric series, log normal, log series, or broken stick models. We transformed total abundance data into 0/1 matrices as input into the program EstimateS We also used EstimateS to calculate Morisita-Horn, and the newly developed Chao-Jaccard and Chao-Sørensen abundance based beta similarity estimators We can consider our samples as real (independent) replicates of the studied sites because repetitive sampling was done independently from one sample to the next and the biological system under consideration is dynamic . This assumption not only validates ANOSIM analysis We used CCA as a constrained ordination direct gradient analysis method to relate RSTs grouped into OTUs to the environmental variables measured. We used CANOCO 4.5 Text S1Environmental tag sequencing methods facilitate comprehensive microbial ecology and biogeography studies.(0.05 MB DOC)Click here for additional data file.


In 1997, an outbreak of human tularemia associated with hare-hunting in central Spain affected 585 patients. We describe the identification of Francisella tularensis biovar palaearctica in a second outbreak of ulceroglandular tularemia associated with crayfish (Procambarus clarkii) fishing in a contaminated freshwater stream distant from the hare-associated outbreak. The second outbreak occurred 1 year after the first.


We speculate that multiple pleural metastasis may disturb the normalrepair mechanism of the lung tissue and cause prolonged persistent air leaks.A rare case of bilateral pneumothorax in a 54-year-old woman with advanced breast cancerassociated with lung and pleural metastases is presented. The patient was admitted to ourhospital complaining of unexpected severe dyspnea. A chest X-ray showed bilateral pneumothoraxassociated with multiple lung metastases and pleural effusions, followed byimmediate pleural drainage. Although air leak and effusions of the right lung were wellcontrolled by the conservative management, massive air leaks of the left lung had continuedfor 40 days. Because of patient's poor general status a surgical closure of the leaking sitewas selected using video-assisted thoracoscopic surgery techniques. Thoracoscopy revealeda ruptured bulla in the lower lobe (S


Lambda tomography (LT) is to reconstruct a gradient-like image ofan object only from local projection data. It is potentially animportant technology for medical X-ray computed tomography (CT) ata reduced radiation dose. In this paper, we prove the firstgeneral formula for exact and efficient fan-beam LT from datacollected along any smooth curve based on even and odd dataextensions. As a result, an LT image can be reconstructed withoutinvolving any data extension. This implies that structures outsidea scanning trajectory do not affect the exact reconstruction ofpoints inside the trajectory even if the data may be measuredthrough the outside features. The algorithm is simulated in acollinear coordinate system. The results support our theoreticalanalysis.


Fifteen patients with acute myelogenous leukaemia were studied to determine if their remission blood leucocytes could be stimulated into taking up [3H] thymidine after in vitro culture with their own cryo-preserved irradiated AML leukaemia cells. In 6/15 patients it was possible to show autologous recognition and equal recognition of their stored leukaemia cells, even when they had previously been maintained in in vitro proliferative cultures in liquid suspension and undergoing myeloid maturation for one week. After in vitro proliferative culture, 4 populations of leukaemia cells produced material in the supernatant media between 3 and 7 days capable of inducing [3H] thymidine uptake in autologous  and allogeneic  AML remission lymphocytes, but not in normal donor lymphocytes. The relevance of these observations to tumour-associated AML antigen is discussed.


Methylmercury compounds were kept intact without decomposition or losses by evaporation. Quantitative recoveries of total mercury were achieved with a two-step microwave attack using a combination of HNO3 and H202 solutions as extractant. The whole pretreatment procedure only takes 15 min, which can be further shortened by an automated robust operation with an open focused system. These analytical procedures were validated by the analysis of environmental certified reference materials. The results confirm that the open focused microwave technique is a promising tool for solid sample preparation in analytical and environmental chemistry.This paper describes rapid, simple microwave-assisted leaching/ digestion procedures for total and mercury species determination in sediment samples and biomaterials. An open focused microwavesystem allowed the sample preparation time to be dramatically reduced to only 24 min when a power of 40-80 W was applied. Quantitative leaching of methylmercury from sediments by HNO