Adjuvant psychological therapy (APT) is a newly developed cognitive behavioural treatment which has been designed specifically to improve the quality of life of cancer patients by alleviating emotional distress and inducing a fighting spirit. We report a phase I/II study which evaluates APT in routine clinical practice. A consecutive series of 44 outpatients with various cancers referred for psychiatric consultation and receiving APT at the Royal Marsden Hospital was studied. Standardised self-report questionnaires were used to measure anxiety, depression and four principal categories of mental adjustment to cancer, namely, fighting spirit, helplessness, anxious preoccupation and fatalism. Statistical comparisons between pre-therapy scores and scores after an average of five APT sessions revealed significant improvement in anxiety, depression, fighting spirit, anxious preoccupation and helplessness. Fatalism scores showed the same trend, but the changes were smaller. Patients with advanced disease showed as much improvement as those with local or locoregional disease. Present results indicate improvement in both psychiatric symptoms and mental adjustment to cancer associated with APT. Whether this association is causal remains to be determined by randomised controlled trials. Such a trial is in progress.


Melospiza melodia) to quantify long-term effects of parental immune experience on offspring immune response. We experimentally vaccinated parents with a novel antigen and tested whether parental vaccination influenced the humoral antibody response mounted by fully grown offspring hatched the following year. Parental vaccination did not influence offspring baseline antibody titres. However, offspring of vaccinated mothers mounted substantially stronger antibody responses than offspring of unvaccinated mothers. Antibody responses did not differ between offspring of vaccinated and unvaccinated fathers. These data demonstrate substantial long-term effects of maternal immune experience on the humoral immune response of fully grown offspring in free-living birds.Knowledge of the causes of variation in host immunity to parasitic infection and the time-scales over which variation persists, is integral to predicting the evolutionary and epidemiological consequences of host–parasite interactions. It is clear that offspring immunity can be influenced by parental immune experience, for example, reflecting transfer of antibodies from mothers to young offspring. However, it is less clear whether such parental effects persist or have functional consequences over longer time-scales, linking a parent's previous immune experience to future immune responsiveness in fully grown offspring. We used free-living song sparrows ( Variation in host immunity to parasitic infection is thought to underpin major evolutionary processes, including host–parasite coevolution, inter-sexual selection and the evolution of sex, and to influence host and parasite population dynamics  to test whether parental exposure to a novel antigen influenced the future humoral immune response of fully grown offspring. During September 2004, we vaccinated a sample of song sparrows with tetanus toxoid. Twelve months later, we tested whether baseline tetanus antibody titre or primary antibody response differed between fully grown offspring of vaccinated and unvaccinated parents. Our use of a novel antigen to assess humoral immunity allows us to distinguish long-term effects of parental vaccination from effects of subsequent natural parasite exposure on offspring immune response  or released without vaccination (n=28). Approximately 40 further colour-ringed individuals were observed but not captured. Then, during September 2005, we measured baseline tetanus antibody titres and the primary antibody response to tetanus vaccination in adult song sparrows and their fully grown offspring that had hatched during 2005 . Briefly, sparrows were mist-netted, blood sampled, vaccinated with 70 μl of tetanus toxoid and released. Individuals were recaptured and blood sampled ca 10 days later. Tetanus antibody titres in baseline and post-vaccination plasma samples were quantified by enzyme-linked immunosorbent assay. Tetanus response was estimated as the difference between post-vaccination and baseline antibody titres. Fieldwork was approved by the University of British Columbia Animal Care Committee.During September 2004, we mist-netted song sparrows on Mandarte as part of a wider study of individual variation in immune response. Captured individuals were vaccinated with 70 μl of tetanus toxoid in the pectoral muscle and released (f), and the period between baseline and post-vaccination blood samples, since these variables influence tetanus response in song sparrows . Eleven mothers (of 21 offspring) and 10 fathers (of 19 offspring) had been vaccinated in 2004. Ten mothers (of 25 offspring) and 13 fathers (of 27 offspring) had not been vaccinated. Vaccinated and unvaccinated parents did not differ with respect to  in 2005 .Across all 46 offspring, baseline antibody titres averaged 7.2±0.7 units and did not differ between offspring whose parents had and had not been vaccinated the previous year . TetanusFicedula hypoleuca) chicks.Previous studies have demonstrated increased antibody titres in eggs or young chicks of parents exposed to specific immune challenges . In songParus major) exposed to nest parasites during laying produced offspring that were more likely to recruit locally , these patterns are not an artefact of selective vaccination-induced mortality in poor quality parents.Maternal antibody transfer may particularly benefit young offspring whose own immune systems have not yet developed, but has also been suggested to permanently alter offspring immune function , but it Such long-term consequences of maternal vaccination may represent direct, permanent effects of maternal antibody transfer on offspring immunology (Although substantial, the increased tetanus response in offspring of vaccinated mothers was considerably lower than the average secondary antibody response measured in 2005 in seven parents that had also been vaccinated in 2004 (4488±664 units, see also


Three intraperitoneal human ovarian cancer xenografts  were used to assess the antitumour activity of intraperitoneal therapy with liposome encapsulated MTP-PE. MTP-PE led to significant prolongation of survival in all three xenograft models, but with varying efficacy. In one tumour model (OS), 80% of mice were cured of tumour by twice weekly therapy for 4 weeks, whereas in another xenograft model (LA), the median survival time was approximately doubled compared to PBS injected and placebo liposome injected controls . The antitumor efficacy of MTP-PE did not correlate with the extent of peritoneal neutrophil infiltration after intraperitoneal therapy. Combined therapy with liposome encapsulated MTP-PE and recombinant murine granulocyte-macrophage colony stimulating factor led to increased survival of mice bearing the LA and HU xenografts, compared to tumour bearing mice treated with either agent singly.


When you're a single-celled organism at the bottom of the food chain, it pays to be resourceful. Diatoms, highly successful photosynthetic plankton responsible for 40% of the net primary production in the oceans, undergo seasonal population explosions called phytoplankton blooms that attract billions of krill, copepods, and other grazing predators. As a defense, wounded diatoms release aldehyde compounds that minimize future diatom casualties by compromising the hatching success of grazers. But these diatom-derived aldehydes can also kill diatoms.In a new study, Assaf Vardi, Chris Bowler, and their colleagues investigated the possibility that the contrasting effects of aldehydes reflect their role as “infochemicals” that trigger different responses attuned to changing conditions in the diatoms' habitat. The authors found that different concentrations of aldehydes produce different diatom responses. At low doses, aldehydes induce resistance to the compound's toxic effects. High aldehyde concentrations, on the other hand, trigger cell death, which may lead to termination of a bloom. Thus, diatom-derived aldehydes regulate the population dynamics of both diatoms and their predators.Thalassiosira weissflogii is a ubiquitous, cosmopolitan species; Phaeodactylum tricornutum is a standard model for understanding diatom biology.To investigate aldehyde effects on diatom cell fate and population dynamics, the authors studied how two cultured diatom species responded to a highly reactive aldehyde called decadienal. Reactive compounds like decadienal are likely to generate a variety of potentially harmful molecules called reactive oxygen species (ROS). But the authors detected increased levels of just one ROS, nitric oxide, an unstable compound involved in a wide range of physiological processes. Monitoring nitric oxide levels with a nitric oxide–sensitive fluorescent dye and time-lapse imaging revealed that both diatom species experienced similar bursts of nitric oxide production about five minutes after decadienal treatment.Treated cells succumbed to decadienal in a time- and dose-dependent manner, with significant increases in fatalities above a specific threshold. Below this threshold, cells survived, but underwent cell cycle arrest. To clarify nitric oxide's role in cell death, the authors stimulated nitric oxide production without using decadienal by using molecules called nitric oxide donors. Next, they pretreated cells with a nitric oxide inhibitor before exposing them to decadienal. The number of dying cells increased along with the levels of nitric oxide in the first experiments, and incidence of decadienal-related cell death decreased with the inhibitor. These results clearly implicate nitric oxide in cell death.P. tricornutum cells that express a calcium-sensitive bioluminescent protein, they tracked changes in intracellular calcium levels in response to aldehydes. As predicted, intracellular calcium levels spiked following decadienal exposure. None of the nitric oxide donors stimulated intracellular calcium production, suggesting that nitric oxide functions downstream of calcium. And, indeed, the nitric oxide synthase that produces nitric oxide was shown to be calcium-activated: after perceiving ambient aldehyde levels, the cell undergoes transient calcium increases that result in nitric oxide production.How does the cell stimulate nitric oxide production? Since plant and animal cells use calcium to perceive a wide range of environmental signals, Vardi et al. reasoned that diatoms might, too. Using Interestingly, nitric oxide production levels varied among the diatoms. Some cells showed rapid increases in nitric oxide production while their neighbors showed delayed responses, suggesting that the signal to produce nitric oxide was propagating through the diatom population. Healthy cells sensed the level of stressed cells in their midst by detecting the wounded cells' aldehyde-generated signal. Cells pretreated with a lower dose of decadienal before receiving a higher dose had far better survival and growth rates than cells treated with only a single high dose. These results suggest that lower decadienal doses may immunize cells, stimulating resistance to normally lethal aldehyde concentrations. This induced resistance may provide diatoms who escape grazing predators with a better chance of surviving the toxic aldehydes released by the dying diatoms.Altogether, these results suggest that decadienal-like aldehydes not only affect the reproductive capacity of grazers but also act as infochemicals that monitor stress levels in diatom populations. During phytoplankton blooms, this stress surveillance system can induce resistance or death. The authors propose that this differential response, regulated by the sophisticated use of intracellular calcium and nitric oxide signals, may determine the fitness and succession of phytoplankton communities.The finding that diatoms use chemical signaling for cell–cell communication provides new insights into the cellular mechanisms mediating biotic interactions at the population level and challenges traditional concepts of phytoplankton bloom dynamics. With thousands of different diatom species potentially producing a diverse array of aldehydes, these reactive compounds may prove to be even more versatile than shown here—and may be a key factor contributing to the ecological success of these organisms. Now researchers can begin to unravel the mechanisms that endow one group of molecules with the means to mediate such diverse responses.


We are pleased to see that our Essay  has sparThere seems to be no disagreement with our main theses—antidepressant advertisements do not accurately represent the evidence base from psychopharmacology, experimental psychiatry, and neuroscience; are not strictly based on the United States Food and Drug Administration (FDA)-approved prescribing label; and may mislead consumers.We believe many have bought into the serotonergic hypothesis of depression/generalized anxiety/social anxiety/obsessive-compulsive disorder/panic disorder/post-traumatic stress/bulimia/premenstrual dysphoric disorder largely because the serotonin reuptake inhibitor (SSRI) medications are licensed for these conditions. We reemphasize that pathophysiology cannot be established through clinical efficacy , yet thiAt the date of this letter, the advertising we presented in our Essay is still widespread, and quite visible on consumer advertising Web sites of SSRI manufacturers.


Chinese hamster ovary (CHO) cells were exposed in vitro to combinations of bleomycin and misonidazole under hypoxic conditions. Only one drug was present at any given time and cells were washed before being exposed to the second drug. Both drugs induced potentially lethal damage (PLD). This damage was repaired under hypoxic conditions very rapidly, and bleomycin-induced PLD was repaired more rapidly than misonidazole-induced PLD. If, after the combined treatment, cells are kept in hypoxia, much of the damage can be repaired.


Airway inflammation increases during acute exacerbations of COPD. Extrinsic factors, such as airway infections, increased air pollution, and intrinsic factors, such as increased oxidative stress and altered immunity may contribute to this increase. The evidence for this and the potential mechanisms by which various aetiological agents increase inflammation during COPD exacerbations is reviewed. The pathophysiologic consequences of increased airway inflammation during COPD exacerbations are also discussed. This review aims to establish a cause and effect relationship between etiological factors of increased airway inflammation and COPD exacerbations based on recently published data. Although it can be speculated that reducing inflammation may prevent and/or treat COPD exacerbations, the existing anti-inflammatory treatments are modestly effective. Exacerbations are a cardinal feature of the natural history of moderate and severe Chronic Obstructive Pulmonary Disease (COPD). PatientTreatment decisions for COPD patients are frequently made according to exacerbation rates. In the ATS/ERS guidelines it is stated that patients with frequent exacerbations should be initiated a trial of inhaled steroids. Systemiwhy inflammation is increased during COPD exacerbations. For practical purposes an attempt is made to categorize aetiological factors of increased inflammation into extrinsic or intrinsic, as it can be seen in table how increased inflammation is associated with the pathophysiology of COPD exacerbations.Although evidence for increased inflammation on COPD exacerbations has been reviewed previously, there has been little focus on why inflammation increases and which the consequences of this increase are,10. InveBacterial infections are generally considered to be the most common causes of COPD exacerbations. It is estimated that more than 40% of all exacerbations are of bacterial origin,11,12. AThe most common bacteria connected to COPD exacerbations are non-typable H. Influenzae, S. Pneumoniae, and M. Cattarhalis,11,12. TAirway bacteria initiate airway inflammation through several interconnecting mechanisms. The surface of bacteria allows the complement system to be activated through the alternative pathway, while specific surface molecules of the bacteria, called Pathogen-Associated Molecular Patterns (PAMPs), bind to pattern recognition receptors on a variety of leukocytes and initiate signalling pathways that lead to the activation of NF-κB and production of proinflammatory cytokines. Once acA significant number of studies in stable COPD patients suggest that airway bacterial infections are associated with increased airway inflammation(18–21). Finding a relationship between bacteria and inflammation on stable COPD adds weight to the argument that bacteria may play a causative role in airway inflammation during COPD exacerbations.Soler et al used protected specimen brush and bronchoalveolar lavage sampling to determine inflammatory cell counts, levels of cytokines concentrations and microbial patterns in stable COPD patients and found that increased neutrophils and tumour necrosis factor-alpha  levels may be related to bronchial colonization. IncreasA large prospective longitudinal study by Sethi et al addressed the hypothesis that patients with bacterial-positive exacerbations show increased inflammation compared to bacterial-negative exacerbations. Among HConsistent with these observations, airway inflammation can be decreased with treatment of the infection. Early evidence came from a relatively small study, which showed that neutrophilic mediators' levels may decrease after treatment of bacterial exacerbations. GompertIn conclusion, three major findings support the hypothesis that bacterial infections are actively implicated in the mechanisms of increased airway inflammation during COPD exacerbations: 1) bacterial infections increase airway inflammation in colonized stable COPD patients 2) bacterial-positive exacerbations show increased inflammation (particularly of neutrophilic type) compared to bacterial-negative exacerbations and 3) eradication of bacteria after a bacterial exacerbation is accompanied by a significant decrease in airway inflammation. A summary of these findings is presented in table Respiratory viruses are important triggers of COPD exacerbations. Initial studies using serology and cell cultures for detecting viral infections suggested that 30% of COPD exacerbations are related to viral infections -30. LatePossible mechanisms of viral-induced inflammation have been described. The airway epithelial cell is the principal host cell for most respiratory viruses. Viral rThere is convincing data that viruses induce inflammation in asthma. In animFurther to these observations, two recent studies showed that viral airway infections during COPD exacerbations are related to airway eosinophilia,40. ThisAtypical pathogens with potential importance in acute exacerbations include M. Pneumoniae, C. Pneumoniae and Legionella spp. Considerable confusion exists in the literature regarding the significance of these potential pathogens in acute exacerbations of COPD. This isin vivo studies on the effect of PM exposure on airway inflammation in COPD patients. There is also lack of information on airway inflammation during PM exposure induced COPD exacerbations, which may be due to difficulties in defining such exacerbations.Epidemiologists have linked ambient particulate air pollution (PM) exposure with exacerbations of pre-existing pulmonary diseases, such as COPD. PM-mediAn imbalance between oxidants and antioxidants may be involved in the development of COPD exacerbations. Almost a decade ago Rahman et al showed that plasma Trolox equivalent antioxidant capacity is decreased in patients presenting with acute exacerbation of COPD. Recent Oxidant stimuli induce cellular expression of inducible nitric oxide synthase and heme-oxygenase-1(HO-1) and increase nitrotyrosine formation. We have shown that there is increased HO-1 expression and nitrotyrosine formation in the airways of COPD patients during severe exacerbations relatively to stable state and that this is accompanied by an increase in indices of neutrophilic inflammation, i.e. neutrophil numbers, MPO and IL-8 levels. EvidencOxidative stress induces the transcription of various inflammatory factors, such as nuclear factor-kappaB (NF-κB) and activator protein-1 (AP-1). It has Activated neutrophils and other inflammatory cells can in turn release reactive oxygen species and increase airway oxidative stress. Because oxidative stress can induce inflammation and vice versa, it is not clear which of the two, oxidative stress or inflammation, is primarily involved in the mechanisms of COPD exacerbations-[the chicken and egg problem].Alterations in innate and adaptive immunity are implicated in COPD pathogenesis. EvidencAccording to our observations changes in lymphocyte subpopulations occur during severe COPD exacerbations. In specAlthough Saetta et al also found increased T cell numbers in endobronchial biopsies from chronic bronchitis patients on mild exacerbations, no difference was detected in CD4 or CD8+ve cell numbers. This diIt would be logical to assume that small increases in airway inflammation in patients with already increased baseline levels of inflammation can easily trigger a COPD exacerbation. This presumes the existence of an inflammatory threshold, above which exacerbation occurs. In relation to this hypothesis, Bhowmik et al showed that COPD patients with frequent exacerbations (≥ 3 episodes/year) have increased baseline sputum IL-6 and IL-8 levels. HoweverOn the contrary, Gombertz et al did not detect any difference in neutrophilic mediators (including IL-8) between frequent (≥ 3 episodes/year) and infrequent exacerbators. InteresIn conclusion, data relating baseline airway inflammation to exacerbation frequency are rather controversial. Part of the existing confusion may be due to significant heterogeneity among COPD patients and to the existence of several factors, like bacterial colonization and bronchiectasis that may increase airway inflammation. In particular, COPD patients with bronchiectasis may represent a distinct group characterized by higher rates of bacterial colonization, increased baseline levels of airway inflammation and longer symptom recovery times at exacerbation.Episodes of COPD exacerbations are characterized by an acute increase in a patient's baseline dyspnoea, cough and/or sputum production. Severe Increased airway inflammation induces many pathologic changes on the airways. Accumulation of inflammatory cells in the airway mucosa by itself causes airway wall thickening. Inflammatory cells can release potentially harmful mediators, such as proteases and reactive oxygen species. NeutropA brief summary of the pathophysiologic events that may link airway inflammation to the symptoms of COPD exacerbations and to respiratory failure is given in figure Increased airway inflammation would be also expected to increase lung tissue oxygen demand and oxygen consumption. In patients with acute lung injury no relation has been found between pulmonary oxygen consumption and lung inflammation, but there is no relevant data in COPD exacerbations.Despite the increasing recognition of the importance of airway inflammation in the development of COPD exacerbations, there is still confusion regarding the role of anti-inflammatory strategies in the prevention and treatment of COPD exacerbations,94. TherOther drugs with anti-inflammatory properties, like methylxanthines and mucolytic agents seem not to be effective on COPD exacerbations. B-agoniIt has been long postulated that airway inflammation may be increased during COPD exacerbations and this may be involved in the pathophysiology of exacerbations. There is now sufficient data to support such a hypothesis. Firstly, there are accumulating observations for increased inflammation during COPD exacerbations. Secondly, specific aetiological factors for this increase have been identified. Thirdly, possible mechanisms that may link airway inflammation with the pathophysiology of exacerbations have been unmasked. Despite significant advances in our understanding of the role of inflammation in COPD exacerbations, the existing anti-inflammatory treatments remain modest and there is little overall benefit for the patient. Further research is needed to target therapies to the appropriate patient populations and to develop new therapeutic strategies.The author(s) declare that they have no competing interests.


Site of first recurrence, disease-free interval (DFI), female sex steroid receptors, ploidy measurements as well as histological grading have been analysed as potentially valuable predictive factors in 313 cases of recurrent breast cancer. Univariate and multivariate analyses show histological grading, site of recurrence and disease free interval to be useful prognostic variables when assessing prognosis once disease has recurred. High concentrations of oestrogen receptors (ER) were found in patients with bone metastases, whereas lower concentrations of ER were related to visceral recurrences. Ploidy measurements failed in this study to give any predictive information once disease recurred.


Internationally there is a 4-fold variation in age-adjusted incidence rates for childhood leukaemia , with only slightly greater worldwide differences specifically for acute lymphocytic leukaemia (ALL) and for acute nonlymphocytic leukaemia (ANLL). Total leukaemia rates are highest among Hispanic populations in Costa Rica and Los Angeles , due primarily to elevated ALL incidence, while low rates occur among US blacks, Kuwaitis, Israeli non-Jews, and Bombay Indians. In most populations the patterns for ALL are similar to those for total leukaema, with peak incidence at ages 1-4 and a decline thereafter. Lower and more uniform rates are generally observed at all ages for ANLL. Age-adjusted rates for ANLL appear to vary substantially among some populations with uniform ALL incidence rates  and yet appear to be similar in other populations with variation in ALL rates . Possible variation among registries in completeness of childhood leukaemia ascertainment and accuracy of diagnosis by cell type should be assessed, while case-control investigations among populations with very high and very low rates may provide useful information about the cell-type specific determinants of childhood leukaemia.


Giardia lamblia has been intensively studied. We have performed a sequence survey project resulting in 2341 expressed sequence tags (EST) corresponding to 853 unique clones, 5275 genome survey sequences (GSS), and eleven finished contigs from the diplomonad fish parasite Spironucleus salmonicida (previously described as S. barkhanus).Comparative genomic studies of the mitochondrion-lacking protist group Diplomonadida (diplomonads) has been lacking, although S. salmonicida from its human parasitic relative G. lamblia were identified such as nineteen putative lineage-specific gene acquisitions, distinct mutational biases and codon usage and distinct polyadenylation signals.The analyses revealed a compact genome with few, if any, introns and very short 3' untranslated regions. Strikingly different patterns of codon usage were observed in genes corresponding to frequently sampled ESTs versus genes poorly sampled, indicating that translational selection is influencing the codon usage of highly expressed genes. Rigorous phylogenomic analyses identified 84 genes – mostly encoding metabolic proteins – that have been acquired by diplomonads or their relatively close ancestors via lateral gene transfer (LGT). Although most acquisitions were from prokaryotes, more than a dozen represent likely transfers of genes between eukaryotic lineages. Many genes that provide novel insights into the genetic basis of the biology and pathogenicity of this parasitic protist were identified including 149 that putatively encode variant-surface cysteine-rich proteins which are candidate virulence factors. A number of genomic properties that distinguish Our results highlight the power of comparative genomic studies to yield insights into the biology of parasitic protists and the evolution of their genomes, and suggest that genetic exchange between distantly-related protist lineages may be occurring at an appreciable rate in eukaryote genome evolution. Giardia lamblia , which is a major cause of water-borne enteric disease in humans in both industrialised and developing countries .The sequences reported here were deposited in GenBank at the National Center for Biotechnology Information [Genbank:JOA co-constructed the second GSS library, finished the sequence of seven contigs, carried out the EST and GSS assemblies, all phylogenetic and most bioinformatic analyses, and drafted and coordinated the editing of the manuscript. ÅMS co-constructed the second GSS library, carried out the majority of the GSS sequencing, and performed initial bioinformatic analyses. DSH grew the organism, purified nucleic acids, constructed the EST library, and carried out some of the initial EST sequencing. CAM constructed the lambda and the initial GSS libraries, carried out the majority of the EST sequencing, the initial GSS sequencing and the sequencing and assembly of three lambda clones. PLD carried out some of the initial EST sequencing. SGS provided advice on the analyses of molecular biological aspects and drafted these parts of the manuscript. RPH performed part of the analyses of cysteine rich and putative mitosomal proteins. JML, MAR, RPH, and AJR co-initiated and supervised the project in their respective laboratories, provided advice on the analyses and edited the manuscript. All authors read and approved the final manuscript.A table showing the characteristics of the finished contigs.Click here for fileComplete list of gene annotations.Click here for fileA table showing the genes putatively involved in LGT events.Click here for filePhylogenetic trees 1–25 for genes putatively involved in LGT events and listed in Additional file 3.Click here for filePhylogenetic trees 26–50 for genes putatively involved in LGT events and listed in Additional file 3.Click here for filePhylogenetic trees 51–72 for genes putatively involved in LGT events and listed in Additional file 3.Click here for file


A world first pineapple EST sequencing program has been undertaken to investigate genes expressed during non-climacteric fruit ripening and the nematode-plant interaction during root infection. Very little is known of how non-climacteric fruit ripening is controlled or of the molecular basis of the nematode-plant interaction. PineappleDB was developed to provide the research community with access to a curated bioinformatics resource housing the fruit, root and nematode infected gall expressed sequences.Arabidopsis homologues, both MIPS based and Gene Ontology functional classifications, and clone distributions. The online resource can be searched by text or by BLAST sequence homology. The data outputs provide comprehensive sequence, bioinformatic and functional classification information.PineappleDB is an online, curated database providing integrated access to annotated expressed sequence tag (EST) data for cDNA clones isolated from pineapple fruit, root, and nematode infected root gall vascular cylinder tissues. The database currently houses over 5600 EST sequences, 3383 contig consensus sequences, and associated bioinformatic data including splice variants, The online pineapple bioinformatic resource provides the research community with access to pineapple fruit and root/gall sequence and bioinformatic data in a user-friendly format. The search tools enable efficient data mining and present a wide spectrum of bioinformatic and functional classification information. PineappleDB will be of broad appeal to researchers investigating pineapple genetics, non-climacteric fruit ripening, root-knot nematode infection, crassulacean acid metabolism and alternative RNA splicing in plants. Ananas comosus (L.) Merrill] is the third most important tropical fruit after banana and mango. Pineapple fruits are classified as non-climacteric, as there is no respiratory burst or spike in ethylene production during ripening and exogenous application of ethylene does not rapidly accelerate fruit ripening. Much has been learnt about the control of fruit ripening in climacteric fruit using tomato as a model system. In particular, manipulation of genes involved in the ethylene biosynthetic pathway and a MADS box transcription factor have led to altered ripening characteristics   . Those cContig consensus sequences containing a polyA tail were analyzed for open-reading frames using EditSeq sequence analysis software  and were BLASTX searched against the GenBank nr protein database . Contig Edited clone sequences generally assembled into contigs with 97–100% homology. However, the contig assembly report occasionally revealed incidences where some clones clustered with between 90–97% homology or that some clones did not cluster into existing contigs due to homology somewhat below the 90% threshold. An inspection of these clone sequences and contigs revealed the presence of apparently unspliced intron sequence, and/or the absence of exon sequence in some of the clones. A comparative analysis to other clone sequences within the contig alignment and to homologous protein coding sequences in GenBank verified that 120 clones contain an apparent "mis-splicing" event. The putative splice variant clones containing un-spliced intron sequence and/or missing spliced exon sequence are listed in the pineapple bioinformatic resource. The presence or absence of a putative splice variant is also reported in contig/clone search outputs.Despite precautions to remove nematodes from root tissues prior to library construction, it was anticipated that there would be some contamination of the pineapple gall libraries with nematode derived sequences. All contig consensus sequences containing root and gall EST's were BLASTN searched against the GenBank dbEST and BLASTX searched against the GenBank nr database. Matches to known nematode sequences were manually inspected and 77 contigs identified as containing a putative nematode sequence. All contigs containing putative nematode sequence are listed within the online pineapple bioinformatic resource.Arabidopsis thaliana.The pineapple EST sequencing project was initiated as a first step toward identifying genes involved in and the molecular basis of non-climacteric fruit ripening and the nematode-plant interaction. The online pineapple bioinformatics resource was developed to house EST sequence information and associated bioinformatic data in a user-friendly format. PineappleDB can be freely accessed via the internet, and currently contains BLAST and text search tools to efficiently mine the dataset for clones and contigs of interest. The resulting search outputs contain comprehensive information on the clone and contigs including cloneID, contig number, number of clones in each contig, nearest BLASTX match, accession number of match, length of match, percent similarity, putative annotation, splice variants, MIPS based functional classifications, Gene Ontology classifications, and the distribution of clones from each library fig. . Links aPineappleDB houses the first reported collection of EST sequences isolated from pineapple. PineappleDB will grow as more EST sequence information becomes available. Furthermore, we have initiated a pineapple microarray project and it is anticipated that gene expression data will be incorporated into the online pineapple bioinformatics resource in the future. The EST database will periodically be upgraded as annotation, functional classification, and gene ontology information is updated.The PineappleDB resource can be accessed via j.botella@uq.edu.auContact: Dr. José R Botella at *The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint first authors. RM was responsible for data collection, the bioinformatic pipeline and manuscript preparation. MC developed the online database and undertook batch BLAST processes. JR-K contributed to the fruit EST editing and identification of full length coding sequences. DJF participated in the conception, design and co-ordination of the study and helped complete the manuscript. JRB designed, supervised and coordinated the project.


Foetal acinar components associated with the development of the hamster pancreas have been previously defined with the aid of an antifoetal pancreas serum. In immunohistology this antiserum also stained malignant ductal cells in N-nitrobis (2-oxopropyl) amine (BOP)-induced pancreatic adenocarcinoma. It did not stain adult pancreas structures including acini, ducts and islets of Langerhans. In this study, re-expression of foetal acinar antigens was disclosed before formation of tumours. Adenocarcinomas were not detected by conventional histology before the 24th week following initiation of the chemical treatment. However, staining with the antiserum was observed from the 7th week appearing in the apex of some acini cells having an almost normal histological appearance. Later, foetal acinar expression was frequently associated with evident morphological alterations in acini like dyskaryosis, enlarged cytoplasm or lumina. Staining of ducts with marked atypical epithelium and  of neoplastic ducts was also observed. It was not detected in other pancreatic lesions viz. cystadenomas, mucoid glands and regular hyperplastic ducts. Acinar dedifferentiation as assessed by expression of foetal components preceded formation of tumours in all instances.


The occurrence of different intercellular junctions in epithelial rat skin tumours induced by methylcholanthrene was investigated using thin sections and freeze-fracture replicas examined by electron microscopy. Tumours which appeared first were basal cell carcinomas. Later, different tumours of hair follicle and of sebaceous gland origin were formed. Finally, in the majority of tumours a squamous component evolved. Metastases developed from the squamous carcinomas exclusively. Desmosomes and gap junctions were detected in basal cell carcinomas whereas, in squamous carcinomas, tight junctions were also seen. While all three types of junction were found in the primary squamous tumours, the tumour metastases in lymph nodes and lungs contained only desmosomes.


A monoclonal antibody, H317, has been used for the sensitive and specific detection of placental-type alkaline phosphatase (PLAP) in sera, solubilized tissue extracts and fixed tumour tissue sections from patients representing a variety of ovarian tumours. PLAP was detected in over 30% of these sera and in most solubilized tumour tissue extracts. There was no association between circulating PLAP levels and either tissue extract levels or immunohistological staining of ovarian tumour tissue sections with H317. Nevertheless, immunohistology demonstrated the heterogeneity of cellular localization of PLAP within different tumours, and can often be of value in localizing tumour tissue.


Numbers of intestinal goblet cells containing specific acid mucins were determined in male Sprague-Dawley rats receiving azoxymethane  with or without jejunoileal bypass (JIB). Controls had injections of vehicle and sham bypass. Thirty weeks postoperatively colorectal length and crypt depth were increased by azoxymethane and further increased by JIB. JIB doubled the yield of intestinal tumours (P less than 0.01). Goblet cells containing sulphomucins normally predominated throughout the intestinal tract. Contents of sulphomucins and especially sialomucins were consistently higher in the small bowel and colon of rats receiving azoxymethane alone, but again the highest values were observed in animals with azoxymethane plus JIB. Both small-bowel bypass and azoxymethane stimulate adaptive growth of the colon and small bowel remaining in circuit. Goblet-cell hyperplasia is a feature of this response, and sialomucins are preferentially secreted by the adapting epithelium.


It is suggested that most childhood acute lymphoblastic leukaemias and some other paediatric cancers are chemo-curable because they arise in stem cell populations that are functionally transient, chemosensitive and programmed for apoptosis. Most adult acute leukaemias are chemo-incurable at least in part because they originate in relatively drug resistant stem cells with extensive self-renewal capacity. The latter property in turn increases the probability of clones evolving with multi-drug resistance. Particular mutations may superimpose additional adverse features on leukaemic cells.


In plants, tandem, segmental and whole-genome duplications are prevalent, resulting in large numbers of duplicate loci. Recent studies suggest that duplicate genes diverge predominantly through the partitioning of expression and that breadth of gene expression is related to the rate of gene duplication and protein sequence evolution.Arabidopsis, there are 53 MST genes that form seven distinct subfamilies. We created profile hidden Markov models of each subfamily and searched EST databases representing diverse land plant lineages to address the following questions: 1) Are homologs of each Arabidopsis subfamily present in the earliest land plants? 2) Do expression patterns among subfamilies and individual genes within subfamilies differ across lineages? 3) Has gene duplication within each lineage resulted in lineage-specific expansion patterns? We also looked for correlations between relative EST database representation in Arabidopsis and similarity to orthologs in early lineages.Here, we utilize expressed sequence tag (EST) data to study gene duplication and expression patterns in the monosaccharide transporter (MST) gene family across the land plants. In Arabidopsis, one or a few genes within most subfamilies have much higher EST database representation than others. Most highly represented (broadly expressed) genes in Arabidopsis have best match orthologs in early divergent lineages.Homologs of all seven MST subfamilies were present in land plants at least 400 million years ago. Subfamily expression levels vary across lineages with greater relative expression of the STP, ERD6-like, INT and PLT subfamilies in the vascular plants. In the large EST databases of the moss, gymnosperm, monocot and eudicot lineages, EST contig construction reveals that MST subfamilies have experienced lineage-specific expansions. Large subfamily expansions appear to be due to multiple gene duplications arising from single ancestral genes. In Arabidopsis MST gene family are ancient in land plants and show differential subfamily expression and lineage-specific subfamily expansions. Patterns of gene expression in Arabidopsis and correlation of highly represented genes with best match homologs in early lineages suggests that broadly expressed genes are often highly conserved, and that most genes have more limited expression.The seven subfamilies of the  Large proportions of genes within genomes are members of hierarchical gene families and superfamilies. Gene families appear to evolve through a combination of tandem, segmental and whole genome duplication (polyploidy) events. A number of researchers in the first half of the twentieth century observed relationships between chromosome duplications and morphological variation . In 1970More recent theoretical and empirical work suggests that gene duplicates are retained more frequently than the classical model permits and that new function or expression arises through the processes of neo- and subfunctionalization ,6. In suPlant genomes contain large fractions of duplicate loci due to the frequent occurrence of segmental duplications and polyploidy events. Following a polyploidy event, there is a rapid loss of duplicate loci in the transition to functional diploidy and the remaining duplicate loci undergo rapid functional divergence . Recent +-sugar symporters localized in the plasma membrane (see references below).In this study, we investigate the monosaccharide transporter (MST) gene family in land plants. MSTs are found in all three domains of life, have fundamental importance in carbohydrate flux and are highly conserved across lineages. All MST proteins are characterized by 12 hydrophobic membrane-spanning domains separated by interconnecting cytoplasmic and extracellular loops, with cytoplasmic N- and C-terminal domains . This hiArabidopsis thaliana genome reveals 53 MST genes that cluster into seven subfamilies on phylogenetic analysis  and whisk ferns (Psilotaceae) that form a monophyletic group which is sister to the seed plants . Text file output from the Click here for filehmmbuild component of the HMMer software package.Profile HMMs for each MST subfamily . Text file output from the Click here for filehmmbuild component of the HMMer software package.Profile HMMs for each MST subfamily . Text file output from the Click here for filehmmbuild component of the HMMer software package.Profile HMMs for each MST subfamily . Text file output from the Click here for filehmmbuild component of the HMMer software package.Profile HMMs for each MST subfamily . Text file output from the Click here for filehmmbuild component of the HMMer software package.Profile HMMs for each MST subfamily . Text file output from the Click here for filehmmbuild component of the HMMer software package.Profile HMMs for each MST subfamily . Text file output from the Click here for fileAlignment of MST subfamily profile HMM consensus sequences. Multiple sequence alignment of consensus sequences generated from profile HMMs for each of the seven MST subfamilies. Sequences were aligned using the ClustalW option in the AlignX component of the VectorNTI package. Amino acid residues highlighted in yellow indicate 100% identity across sequences.Click here for fileArabidopsis thaliana EST database.Summary of identified MST ESTs in Click here for fileMarchantia polymorpha, Selaginella lepidophylla, Ceratopteris richardii).Summary of identified MST ESTs in small databases (Click here for filePhyscometrella patens and Physcomitrella patens subsp. patens EST databases.Summary of identified MST ESTs in Click here for filePinus taeda EST database.Summary of identified MST ESTs in Click here for fileZea mays EST databaseSummary of identified MST ESTs in Click here for fileLycopersicon esculentum EST databaseSummary of identified MST ESTs in Click here for file


PCR-based microsatellite polymorphisms have proved their power in genetic linkage analysis and other identification methods, due to their high information content and even distribution over the chromosomes. In the present study we applied microsatellite polymorphisms to detect loss of heterozygosity in fresh (snap-frozen) and in archival ovarian tumour tissue. Clear allele losses were found in fresh and paraffin embedded tumour samples. Conventional Southern analysis of flanking markers on the same tumour DNA samples confirmed the observed losses detected by microsatellite polymorphisms. Titration experiments suggest that loss of heterozygosity remains detectable in tumour samples despite 60% contamination with normal DNA. This technique provides a fast and reproducible alternative to conventional Southern blotting in the detection of loss of heterozygosity, with the crucial additional advantages of minimal sample requirements, making archival material available for genetic investigation.


Human colon adenocarcinoma cells (LoVo) resistant to the new antitumor agent FCE 24517 [benzoyl-mustard derivative of distamycin A] (LoVo/24517) are resistant to the selecting agent and related molecules as well as to vinblastine, with marginal or no resistance to other antitumour drugs. Treatment with verapamil, tamoxifen, nicergoline or cyclosporin A only partially restores the activity of FCE 24517 against LoVo/24517 cells. Such results suggest that resistance mechanisms possible specific for this class of compounds are operating.


From 1,211 breast cancers, 15 oestrogen receptor (ER) negative-progesterone receptor (PgR) positive breast cancers by conventional dextran coated charcoal steroid binding assays in cytosol were reassessed using Elisa techniques with monoclonal antireceptors antibodies in the cytosolic and nuclear fractions, and immunocytochemistry on cryostat sections. Three categories of results were found in this series. Two tumours were false negative ER due to receptor sites occupancy by hormonal contraceptive treatment. A second group of ten tumours, with high PgR concentrations and immunoreactive ER, corresponds to non ER-binding forms of receptors. One PgR positive tumour was found to be devoid of PgR by using monoclonal antiPgR antibodies might contain a progesterone binding cyst protein. Only two tumours were found to be true ER negative-PgR positive by all methods. This rare phenotype deserves further study of the regulation of the PgR gene.


The metastatic sites of infiltrating duct (IDC) and infiltrating lobular carcinoma (ILC) have been compared using both clinical and autopsy data. The following statistically significant differences were found: Lung parenchymal metastases were more common in IDC. Bone trephine biopsies were more likely to be positive in ILC. Carcinomatous meningitis was associated almost exclusively with ILC. Peritoneal/retroperitoneal metastases of distinctive pattern occurred in ILC. There was often associated linitis plastica-like involvement of the stomach wall and diffuse infiltration of the uterus. Hydronephrosis was a common secondary phenomenon.


Mutagenicity and cytotoxicity are basic cellular effects of cigarette smoke which underlie the development of lung cancer and chronic obstructive airways disease. This study reports that, on a weight-for-weight basis, cigarette smoke condensates from low, middle and high tar cigarettes produce similar mutagenic effects detected by induced sister chromatid exchanges and similar cytotoxic effects detected by vital dye exclusion in human leucocytes. These findings, taken with the strong evidence that smokers extract more smoke from lower tar cigarettes to compensate for low nicotine yields, suggest that the health dangers associated with smoking these "safer" products are underestimated.


Mexipyrgus churinceanus) and a molluscivorous cichlid (Herichthys minckleyi), we examined three components of this interaction: 1) spatial variation in two putative defensive traits, crushing resistance and shell pigmentation; 2) whether abiotic variables or frequency of molariform cichlids are associated with spatial patterns of crushing resistance and shell pigmentation and 3) whether variation in primary productivity accounted for small-scale variation in these defensive traits.Recent models suggest that escalating reciprocal selection among antagonistically interacting species is predicted to occur in areas of higher resource productivity. In a putatively coevolved interaction between a freshwater snail  and snail defensive traits. However, crushing resistance and frequency of pigmented shells were negatively correlated with molariform frequency. Crushing resistance and levels of pigmentation were significantly higher in habitats dominated by aquatic macrophytes, and both traits are phenotypically correlated.M. churinceanus exhibit striking variation at small spatial scales often associated with differences in primary productivity, substrate coloration and the frequency of molariform cichlids. These local geographic differences may result from among-habitat variation in how resource productivity interacts to promote escalation in prey defenses.Crushing resistance and pigmentation of  Mexipyrgus churinceanus.Spatial variation in defensive traits is common in many antagonistic interactions -3. The gMexipyrgus churinceanus as well as its polymorphic fish predator Herichthys minckleyi are endemic to the isolated Cuatro Ciénegas valley in the Mexican Chihuahuan desert . Increased resource availability probably allows greater investment in costly shell material, leading to the prediction that hotspots may occur in areas with greater food resources for snails.Alternatively, variation in shell strength might be primarily a response to biotic influences. We address whether spatial variation in snail defenses is associated with two biotic factors: molariform frequency and resource productivity. If molariform H. minckleyi, and this divergence occurs at very small spatial scales in a geographically mosaic fashion, we predict there will be no significant positive autocorrelation at small spatial and genetic distances. In the current study, we determine genetic distances between snail populations from a previously published study of mtDNA sequence variation in Mexipyrgus [A critical assumption of these population-based measures of phenotypic divergence is that they are statistically independent. This assumption may be violated because nearby snail populations may experience similar predation pressure because cichlids are more mobile than these brooding snails. To address this issue we assess whether the similarity of shell defensive traits among nearby populations can be attributed to either geographic proximity or genetic similarity -25. Genexipyrgus .Mexipyrgus. First, we examine spatial variation in two putative defensive traits, crushing resistance and shell pigmentation. Then, we tested whether abiotic or biotic variables account for spatial patterns of crushing resistance and shell pigmentation. Finally, we determine whether variation in primary productivity accounts for small-scale variation in these defensive traits.We address three questions concerning the evolution of defenses in 2 = 43.9%, p < 0.0001), whereas shell shape explained little variation in crushing resistance . We next used analysis of covariance to examine the effect of population on crushing resistance using size as a covariate. The interaction term (shell size by population) was not significant (p = 0.24), and there was a highly significant effect of population on size-adjusted crushing resistance . There was considerable spatial variation in size-adjusted crushing resistance, ranging from 42.3 Newtons in a Rio Mesquites population to 92.3 Newtons in a Tierra Blanca population , indicating the absence of autocorrelation among populations at any scale.Mean and standard errors of crushing resistance and shell size are presented in Table ion Fig. . The cor2 = 310.7, n = 19, p < 0.001). The correlograms of pigmentation frequency and all distance measures were not significant (see figure b distance classes), again indicating the absence of autocorrelation among populations at any scale.There was considerable variation in the frequency of pigmentation among populations Fig.  and therp = 0.19) or conductivity . Similarly, there was no significant correlation between pigmentation frequency and temperature  or conductivity . In contrast, there were significant negative correlations between molariform frequency and both size-adjusted crushing resistance .There was no significant correlation between size-adjusted crushing and temperature , crushing resistance was higher where Nymphaea was abundant . Similarly, RM1 had significantly higher crushing resistance than nearby RM3 . The Los Remojos populations did not differ significantly in size-adjusted crushing resistance . In the paired adjacent populations, the frequency of pigmented shells was significantly higher in darker versus lighter substrates in all 3 comparisons: Rio Mesquites 1 and 3 ; Los Remojos N and S ; and Tio Candido N and S .In the two of the three paired adjacent populations that differ in t Figure . For exaMexipyrgus churinceanus in Cuatro Ciénegas and the presence of the molariform cichlids are suggestive of coevolutionary selection [Pronounced geographic differences in prey defenses are probably common ,27. Whilelection ,15, the Mexipyrgus churinceanus requires an assessment of phenotypic variation and the spatial autocorrelation of these traits among populations at small genetic and geographic distances. There is significant spatial variation in M. churinceanus crushing resistance and pigmentation at small spatial scales suggesting that certain factors may cause this mosaic distribution of snail defensive traits. In paired populations separated by very small linear and genetic distances, elevated crushing resistance occurs in habitats containing extensive Nymphaea stands. Crushing resistance is dramatically lower in light colored substrates lacking Nymphaea. Because Tio Candido and Rio Mesquites populations represent distinct lineages based on mtDNA sequence differentiation [Mexipyrgus feeds extensively . Increased resource availability likely allows greater investment in costly shell material, and experiments that manipulate resource availability to test its effect on Mexipyrgus shell strength would provide a further test of this hypothesis.To address whether prey defenses have evolved independently across the geographic range of ntiation , it suggCamellia host plant indicate that escalation in armaments only occurred in southern latitudes [Mexipyrgus is the result of coevolutionary interactions between endemic snails and the molluscivorous morph of H. minckleyi [Nymphaea habitats, increased snail crushing resistance may reduce molariform fitness due to increased costs of crushing, while papilliform fitness is higher due to increased availability of plant material for shredding. We plan to experimentally test whether these alternative cichlid morphs have fitness trade-offs in different resource environments, and more quantitatively measure resources available to Mexipyrgus.Although there is considerable documentation of hotspots and coldspots in coevolved antagonistic interactions, how spatial processes and community composition promote coevolution and generate selection mosaics has received less attention. A recent study of herbivorous weevils and their atitudes . More noinckleyi . SurprisMexipyrgus shell pigmentation is also associated with different substrate coloration and there is no evidence of genetic or spatial autocorrelation among populations. In the three geographically-paired populations, snails with pigmented bands were significantly more common in Nymphaea habitats and unbanded snails were more common where the benthic substrate was light and marled- colored. Unbanded snails are probably more cryptic against lighter benthic substrates and banded snails are more cryptic against the darker substrates found in association with Nymphaea. This type of background matching by prey under selection from highly visual predators is common in many organisms [Microspatial variation in rganisms -30 and eNymphaea is absent, investment in crushing resistance may be constrained due to resource limitation, so that crypsis is the most effective defense against molariform predation. We suspect that investment in shell pigmentation is not as costly as having more robust shells, and that snails are capable of inexpensively modifying their pigmentation in Nymphaea habitats to increase crypsis. Whether correlational selection acts on pigmentation patterns and shell strength in these habitats requires a deeper understanding of the phenotypic plasticity in both traits. One hypothesis is that increased pigmentation in these environments does not increase background matching, and pigmentation only increases due to correlational selection on crushing resistance. This hypothesis could be easily refuted if predation success on banded shells is less in Nymphaea habitats. Given evidence for inducible defenses in snails [The positive relationship between the two snail traits across environments suggest that pigmentation and crushing resistance may represent phenotypically-correlated traits. In resource-poor environments where n snails -33, we aH. minckleyi undoubtedly use their jaws to crush snails, Cuatro Ciénegas may represent an ideal system for investigating mosaic predator-prey coevolution. In future studies, we plan to examine local adaptation of molariforms, escalation of molariform traits in resource rich environments, and heritability and phenotypic plasticity of both crushing resistance and pigmentation across resource gradients.Spatial variation in two prey defense traits, crushing resistance and shell pigmentation, exhibits striking variation at small spatial scales often associated with habitat differences in primary productivity and substrate coloration. These local geographic differences may result from among-habitat variation in how resource productivity interacts to promote escalation in prey defenses. Because molariform Mexipyrgus churinceanus from various drainages throughout the entire Cuatro Ciénegas basin , and whether these correlations change as a function of geographic and/or genetic distance. Under the spatial mosaic hypothesis, we predicted no positive autocorrelations of geographically-adjacent or genetically-similar populations. We estimated Moran's I using Passage . Moran'sariation  using a To assess the potential influence of abiotic factors on shell strength, we measured temperature and conductivity in the 19 populations . Conductivity measures were temperature compensated. Above ground flow connects a few of the populations examined here, but the abiotic factors of each are influenced by large discharge from separate isothermal springs . Therefoet al [To estimate the relationship between frequency of molariforms and both size-adjusted crushing resistance and frequency of pigmented shells, the frequency of molariforms in ten populations was obtained from two sources. The frequency in Mojarral Oeste, North Tio Candido, Tio Candido, Los Remojos Negro, and Los Remojos Blanco was estimated in 2001 by Kloeppel , and waset al . Fish weNymphaea is associated with variation in snail crushing resistance and pigmentation, we compared the crushing resistance  and frequency of banded snails (Log likelihood tests) in three pairs of adjacent populations in the Rio Mesquites, Los Remojos, and Tio Candido. These paired populations are in very close proximity to one another . In these paired populations, substrate coloration differed dramatically due to the presence or absence of Nymphaea beds. We also examined the phenotypic correlation between mean population estimates of the number of bands and crushing resistance. We conducted all statistical analyses in SPSS [To address whether the presence or absence of  in SPSS .All authors conceived the study and participated in its design and conducted field work. SGJ performed statistical analysis and drafted the manuscript. CDH and FJGL helped with revisions of the manuscript. All authors read and approved the final manuscript.


As animal extinctions continue at the rate of one every 16 years, it's unclear how declining biodiversity will disturb ecosystem dynamics. Of special concern are the pollinators, essential players in the reproductive biology of plants, the earth's primary producers. Millions of years of evolutionary coadaptations lie behind the perfect pairing of pollinator proboscis anatomy with plant flower structure, as well as the mechanisms plants use to attract reproductive assistants to their food rewards. Agave plants emit musky aromas that attract lesser long-nosed bats to nectar stores within their flowers, for example. As the bats travel from flower to flower, pollen collects and then falls from their fur, facilitating cross-pollination.These mutually beneficial relationships are sometimes so specialized that the loss of one species threatens the existence of the other, raising troubling questions about the likely consequences of declining diversity in pollination networks. In a new study, Colin Fontaine et al. tackled this question by experimentally manipulating plant and pollinator interactions under natural conditions. The authors found strong functional relationships between different pollinators and plant communities, with the highest plant community sustainability associated with the most diverse group of pollinators. These findings suggest that loss of biodiversity in pollination networks may threaten the persistence of plant communities.For their study, the authors chose plants with easy and harder access to food rewards—three open-flower and three tubular-flower species—and insects with short and longer mouthparts—three syrphid fly and three bumblebee species. In the spring of 2003, Fontaine et al. set up 36 plant communities in nylon-mesh enclosures in a meadow 80 kilometers (about 50 miles) southwest of Paris, after sterilizing the soil to destroy seeds and pathogens. They planted 30 adult plants in each plot at the same density, and then captured and released local pollinators into the cages during the flowering season . To test all the possible plant–pollinator combinations, the authors set up three plant treatments , then applied three pollination treatments  to each plant treatment.A month after the first pollination treatments, the authors tallied all the fruit on each plant, then randomly selected five fruits per plant (excepting one plant species from each group that would have required harvesting the fruit) to estimate seed production per plant. During the seedling season, they totaled the plants and the seedlings to measure plant population and reproductive success. Pollinator identity determined fruit production, with bee-pollinated plants most productive, and the different plant groups responded differently to the two pollinator groups. As expected, short-mouthed syrphid flies pollinated only open flowers, while bees pollinated both plant types. As a result, tubular flowers produced far fewer fruits with syrphid pollinators while open-flower fruit production remained the same regardless of pollinator. Fruit production increased along with both plant and pollinator diversity. Seed production was a bit more complicated. Though bee-pollinated open flowers produced fewer seeds per plant than those pollinated by syrphids, higher fruit production compensated by producing more seedlings. Fruit production increased with pollinator diversity. As for long-term effects on plant reproductive capacity and success, tubular plant communities had fewer plants at the seedling stage than openflowered plants (and even fewer when pollinated by syrphids). The plant species number and total plant number increased when both pollinator groups were present, and were highest with maximum plant and pollinator diversity. Seedling production showed a similar pattern: mixed plant communities treated with both pollinators yielded the most seeds.What happened? Not surprisingly, the pollinators stuck to their preferred plant: syrphids visited mostly open flowers, and bees visited mostly tubular flowers. Bees can pollinate open flowers but prefer tubular flowers when they have the choice, suggesting that bees may not fill a void left by a different pollinator. The presence of both pollinators allowed more appropriate pairings between insects and flowers—each performing a complementary role—leading to increased pollination efficiency and plant reproductive success.Liza GrossWhile the study offers an admittedly pared down view of pollination networks, it demonstrates the value of studying the functional effects of pollination networks in the field. These results show that losing a species affects plant–pollinator communities, and that such losses may ultimately trigger further reductions in biodiversity, possibly reverberating through the food chain. With as many as 70% of plant species dependent on animal pollinators and at least 82 mammalian pollinator species and 103 bird pollinator species considered threatened or extinct, this is sobering news. —


James and Lange proposed that emotions are the perception of physiological reactions. Two-level theories of emotion extend this model to suggest that cognitive interpretations of physiological changes shape self-reported emotions. Correspondingly false physiological feedback of evoked or tonic bodily responses can alter emotional attributions. Moreover, anxiety states are proposed to arise from detection of mismatch between actual and anticipated states of physiological arousal. However, the neural underpinnings of these phenomena previously have not been examined.We undertook a functional brain imaging (fMRI) experiment to investigate how both primary and second-order levels of physiological (viscerosensory) representation impact on the processing of external emotional cues. 12 participants were scanned while judging face stimuli during both exercise and non-exercise conditions in the context of true and false auditory feedback of tonic heart rate. We observed that the perceived emotional intensity/salience of neutral faces was enhanced by false feedback of increased heart rate. Regional changes in neural activity corresponding to this behavioural interaction were observed within included right anterior insula, bilateral mid insula, and amygdala. In addition, right anterior insula activity was enhanced during by asynchronous relative to synchronous cardiac feedback even with no change in perceived or actual heart rate suggesting this region serves as a comparator to detect physiological mismatches. Finally, BOLD activity within right anterior insula and amygdala predicted the corresponding changes in perceived intensity ratings at both a group and an individual level.Our findings identify the neural substrates supporting behavioural effects of false physiological feedback, and highlight mechanisms that underlie subjective anxiety states, including the importance of the right anterior insula in guiding second-order “cognitive” representations of bodily arousal state. Recent theory distinguishes between two levels of emotional experience: phenomenology and awareness The importance of second-level appraisal mechanisms to emotional judgments is highlighted by false feedback experiments Recent studies of interoceptive awareness In the present study, we examined neural activity associated with effects of false feedback of arousal state (heart rate) on emotional evaluations. Our approach differed from that of Valens We first examined the influence of physiological feedback  on behavioural ratings of intensity of face stimuli  in a repeated measures ANOVA. We observed a significant multivariate interaction of expression type and feedback on attributed emotional intensity , in addition to a significant main effect of facial expression . Planned contrasts revealed a significant interaction between expression  and feedback  . Further, the emotional intensity of neutral faces was rated significantly higher during false relative to true feedback . Across all conditions, the emotional intensity of neutral expressions was rated significantly lower than either angry  or happy expressions  see We next examined the influence of isometric exercise in a repeated measures ANOVA, including the factors exercise (present absent) emotion  and feedback . Exercise did not alter behavioural ratings of emotional expressions, and did not differentially influence false feedback effects.2 = 0.751] diastolic  and mean arterial pressure , stroke volume  cardiac output  and heart rate  relative to the no-exercise condition  and exercise  isometric exercise was associated with significant increases in systolic . This interaction was observed in the right anterior insula cortex  and alsoWe next examined evidence for a cardiac synchronous/asynchronous comparator mismatch function for the anterior insula cortex during interoceptive demand. Predicated on the on the notion that interoceptive information has greater salience when processing ambiguous  stimuli, we hypothesised that this would enable us to test for a comparator function of insula cortex in responding to mismatches in cognitive and interoceptive representations of physiological state, even at conditions of low bodily arousal. Thus, we specifically tested for a greater difference between asynchronous and synchronous cardiac feedback during the processing of neutral expressions, relative to the same difference during the processing of either happy or angry expressions; i.e. [ > ]. Significant interactions were observed in three regions; bilateral anterior insula cortex and dorsal cerebellum .As detailed above, exercise did not alter behavioural ratings of facial expressions-and showed no interaction with false feedback when rating neutral expressions. As a consequence we did not focus our investigations on exploration of neural correlates of non-significant behavioural interactions. Nevertheless, mapping of changes in first level representation within consciously accessible interceptive cortices, notably right anterior insula, was of direct interest.In both exercise and rest conditions, the pattern of activity reflecting the influence of feedback veracity on rating of facial expressions was maintained, with significant interactions observed within right amygdala, bilateral posterior insula and right anterior insula. Within the right anterior insula cortex, the mean BOLD signal change in response to face stimuli portraying neutral expressions predicted individual differences in the magnitude of intensity ratings of these stimuli in the context of false feedback .Pursuing the relationship between cerebral activity and false feedback induced biasing of emotional judgements, we used parametric analyses to examine within each participant the relationship between trial-by–trial behavioural ratings of neutral faces and regional BOLD activity. Again, within the right anterior insula cortex, we observed a significant association between BOLD activity and attributed intensity of neutral expressions during false feedback  . IncreasThe present study demonstrates the influence of false feedback of physiological arousal on emotional appraisal of facial expressions through engagement of a discrete set of brain regions implicated in social and motivational behaviour. This neuroanatomical matrix encompasses brain regions that have been implicated functionally in the encoding of external emotional cues , representation of internal visceral information (insula cortex) and the contextual synthesis of emotional information for declarative awareness (anterior insula/operculum).Our findings suggest a central integrative role for the anterior insula cortex. This region mapped differential influences of false feedback of arousal on differential processing of emotion, was sensitive to the timing of cardiac feedback  and predicted within and across participants the degree to which behavioural ratings were altered during false physiological feedback. Together these novel data reveal neural substrates supporting the second level appraisal of emotional information, arising from an integration of perceptual processing within the context of physiological feedback representations, and influencing subjective affective judgement. Previous studies report right anterior insula engagement during conscious processing of emotional stimuli and in response to evoked physiological arousal responses False feedback of increased cardiac rate during rest enhanced the perceived intensity of neutral facial expressions but not angry or happy facial expressions. This is consistent with previous findings within the false feedback literature that the influence of physiological feedback is greatest during ambiguous judgements Anatomically, Craig argues that the specialised contribution of right anterior insula in conscious interoception derives from a remapping of mid and posterior viscerosensory representations We anticipated a contribution from amygdala to the emotional appraisal task. Patients with selective amygdala lesions show impairments in general social behaviour that correlate with deficits in emotional judgments of face stimuli Behaviourally, false physiological feedback enhances attention to stimuli presumed to be physiologically arousing Overall, we report the modulation of intensity judgements of face stimuli, particularly neutral faces by false feedback of physiological arousal state. Our neuroimaging data indicates engagement and interaction of three systems governing; first, initial representation and homoeostatic control of bodily arousal (insula); second, ‘automatic’ encoding of emotionally salient events  and; third, visual and cognitive appraisal of stimuli . Further, our observations suggest a critical role for right anterior insula cortex in integrating information across these systems to support a second-level representation of emotionality that underpins subjective and behavioural experiences arising from false feedback. We did not observe any direct influence of first level physiological arousal on intensity judgements in the current study despite inducing significant increases in cardiovascular responses through isometric exercise. It is likely that misattribution of arousal is most probable where participants are unaware of the cause of their physiological arousal (i.e. excitation transfer effects). We did not observe misattributed arousal in the present study, presumably because first-level physiological arousal could be correctly attributed to isometric exercise.The present study represents the first investigation of neural responses during both false feedback and exercise induced arousal. We observed an interaction of emotion and feedback on subjective measures, and a parallel interaction within regions implicated in cognitive appraisal, emotional processing and interoception. Our findings suggest an integrative contribution of right anterior insula cortex in second level representations of emotion that, in turn, predict individual differences in emotional behaviour. In sum, our findings provide insight into emotional appraisal mechanisms proposed in two-level theoretical models of emotion and identify the functional neuroanatomical substrate for hierarchical emotional appraisals of the social brain.Twelve healthy right handed individuals  gave informed written consent to take part in this study which was approved by the joint ethics committee of the Institute of Neurology and the National Hospital for Neurology and Neurosurgery, Queen Square, UK. Participants were screened to exclude psychiatric or systemic medical disorders and current medication usage.Before scanning, each participant was familiarised with the task and procedures. Participants were informed that we were interested in the effects of different facial expressions on heart rate during exercise and rest. Individual heart beats were recorded via a pulse oximeter linked to the task computer and were audible as tones (pulses) through headphones. Participants were also informed that the tones  were essentially irrelevant, but were generated to allow us to measure heart responses. During questioning after the study, no participant said that this cover story raised suspicions.true feedback, tones were presented synchronous with individual heartbeats. During asynchronous feedback, tones were delayed by half the inter-beat interval. During false feedback, no-exercise tones were presented at a rate ten percent faster than the preceding exercise heart rate, whereas exercise tones were presented at a rate 10 percent slower than the preceding no-exercise heart rate. Afterwards, participants completed another three minutes of the emotional appraisal task in the alternative exercise condition (either exercise or no-exercise) and with all three feedback conditions. The order of feedback and exercise conditions was randomized. In sessions where a no-exercise condition followed an exercise condition, a 90-second pause followed the instruction to relax grip, to allow exercise associated arousal to decline. At debriefing, participants were asked about their perceptions of heartbeat feedback. All participants were unaware (i.e. did not declare on direct questioning) of the manipulations of the auditory feedback of their heart rate . Further, participants confirmed that they remained focused on the face rating task.During the experimental task , a compu3 T1-weighted structural scans were acquired from each participant.Functional echo-planar datasets sensitive to BOLD (Blood Oxygen Level Dependent) contrast were acquired at 1.5 Tesla (Siemens Sonata) The sequence, minimizing orbitofrontal signal dropout http://www.fil.ion.ucl.ac.uk/spm/), employing spatial realignment and sequential co-registration . Structural scans were segmented into CSF, grey and white matter images and iteratively normalized to standard space  using a single generative model Images were pre-processed using SPM5  of discrete contrasts within the general linear model. Subsequent second-level group random effects analyses were performed on the SPM contrast images of first level canonical HRF responses to permit formal inferences about population effects Peripheral physiological responses were also explored outside the scanner in an independent group of eight healthy right handed participants  because recording equipment was not compatible with the fMRI environment. Participants were given identical task instructions and completed the experimental task while a finger cuff recorded beat-to-beat blood pressure with a Finometer . This allowed a reconstruction of brachial artery pressure from cardiological measures were obtained, including arterial pressure at systole and diastole, stroke volume, cardiac output, left ventricular ejection time, total systemic peripheral resistance, and heart rate.


I read the paper by Chi et al. with interest. The authors found a greater hypoxia response gene expression in carcinomas than stromal cells grown in vitro, but were not sure of the underlying explanation (see Discussion in ). The unThe differences in mitotic activity could also explain greater hypoxia-related gene expression in clear-cell carcinoma compared with chromophobe carcinoma, normal tissue, or oncocytoma, as clear-cell carcinomas are more active mitotically. Did the authors compare the hypoxia gene expression with mitotic activity in various tumors?


Many biological problems such as the detection of co-expressed genes, co-regulated genes, and transcription factor binding motifs rely heavily on the analyses of these image patterns. The increasing availability of ISH image data motivates the development of automated computational approaches to the analysis of gene expression patterns.Staining the We have developed algorithms and associated software that extracts a feature representation of a gene expression pattern from an ISH image, that clusters genes sharing the same spatio-temporal pattern of expression, that suggests transcription factor binding (TFB) site motifs for genes that appear to be co-regulated (based on the clustering), and that automatically identifies the anatomical regions that express a gene given a training set of annotations. In fact, we developed three different feature representations, based on Gaussian Mixture Models (GMM), Principal Component Analysis (PCA), and wavelet functions, each having different merits with respect to the tasks above. For clustering image patterns, we developed a minimum spanning tree method (MSTCUT), and for proposing TFB sites we used standard motif finders on clustered/co-expressed genes with the added twist of requiring conservation across the genomes of 8 related fly species. Lastly, we trained a suite of binary-classifiers, one for each anatomical annotation term in a controlled vocabulary or ontology that operate on the wavelet feature representation. We report the results of applying these methods to the Berkeley Drosophila Genome Project (BDGP) gene expression database.Our automatic image analysis methods recapitulate known co-regulated genes and give correct developmental-stage classifications with 99+% accuracy, despite variations in morphology, orientation, and focal plane suggesting that these techniques form a set of useful tools for the large-scale computational analysis of fly embryonic gene expression patterns. This technique localizes specific mRNA sequences in tissues/cells by hybridizing a labeled complimentary nucleotide probe to the sequence of interest in fixed tissues. Visualizing the probe by colorimetric or fluorescent microscopy allows for the production of high quality images recording the spatial location and intensity of gene expression.A large body of work analyzing DNA micro-array data from microorganisms has demonstrated the value of gene expression analysis in understanding gene function and dissecting gene regulation -3. Whilein situ databases, such as the Berkeley Drosophila Genome Project (BDGP) gene expression pattern database , sh, shin siIn Figure In regard to the automatic annotation of gene expression patterns, we also collected statistics of the use of the ontology annotation terms (results not shown). The percentage of genes corresponding a common annotation term ranges from less than 1% to about 20%. Given this small fraction, it is unlikely that our image-based gene clustering results in Figures Our image analysis methods can be applied to 3D gene expression patterns or other types of aligned image patterns in different contexts. For example, in  we definin situ hybridization brain images of 20,000 genes were aligned to a standard reference atlas. The effective clustering and recognition of these gene expression patterns, based on various image features , could in situ fly gene expression patterns. We have successfully extracted useful local and global image features, and used these to automatically cluster and annotate gene expression patterns with high reliability. Our techniques provide useful tools for the large-scale computational screening of fly embryonic gene expression patterns, as well as the aligned image patterns for similar problems in other model systems.We have developed a set of automatic image analysis methods for For the BDGP database, 2D embryonic images of gene expression patterns were acquired using a digital camera. In a typical image, a single embryo resides in the central part of the image and presents a lateral view. Only lateral views were used, but the embryo can otherwise have an arbitrary orientation. As the embryonic region has much richer texture information than the image background, the embryo can be segmented by thresholding the local variance of a small region (e.g. 3 × 3 pixels) around each pixel. The pixel is binarized to "foreground" if the variance is larger than a predefined threshold (e.g. 2), otherwise to "background". Binarization classifies most embryo pixels as "foreground" and most background pixels as "background", thus producing a mask image that essentially captures the embryonic region. For a segmented embryo, we computed the principal direction along which the variation of all embryonic pixel-coordinates is the greatest and considered this the anterior-posterior axis of the embryo. We then rotated the image to make this axis horizontal. Finally, the embryonic region was cropped and its size was standardized to 400 pixels wide and 200 pixels high.in situ gene expression images in the BDGP database for 1,700 fly genes. We found that about 67% of the images have only one embryo region in the center, and can be easily segmented based on thresholding the pixel variance. These 20,000 extracted image patterns were automatically rotated so that their longest axes are horizontal. We developed a web-based image pattern browser at [We processed about 30,000 owser at , which cowser at .In analyzing the data, we further ignored all images that were not of a lateral view of the embryo. While most images are lateral views, a significant fraction is taken from difference vantage points, such as along the dorsal/ventral axis or some tilted angle. From these viewpoints it is especially difficult to understand the 3D pattern as the embryo is clear and one is essentially seeing the 2D projection of the stain along the viewing axis. It remains an open problem how to effectively use such additional data.For the experimental results reported, we focused on a set of 456 genes. We separated the lateral and dorsal views manually and also adjusted the orientations of these images to assure these images are compared in the correct way, i.e. anterior is at the left and dorsal is up. If in a particular stage-range a gene has multiple images, our computer program merged these images and used their mean-image as the "representative" for this gene. In this way, the image clustering and annotation algorithms would not be biased by the image-numbers of genes. Due to the great variation of the quality of the BDGP 2D image patterns, these processing steps were necessary to produce meaningful results.Abbreviations of the anatomical annotations used throughout the paper:AM amnioserosaAAISN amnioserosa anlage in statu nascendiAISN anlage in statu nascendiAEA anterior endoderm anlageAEAISN anterior endoderm anlage in statu nascendiAEP anterior endoderm primordiumAMP anterior midgut primordiumCMP cardiac mesoderm primordiumCB cellular blastodermCLP clypeo-labral primordiumDEA dorsal ectoderm anlageDEAISN dorsal ectoderm anlage in statu nascendiDECP dorsal ectoderm primordiumDEDP dorsal epidermis primordiumDPMP dorsal pharyngeal muscle primordiumEAISN endoderm anlage in statu nascendiECBG embryonic central brain gliaECBN embryonic central brain neuronECNS embryonic central nervous systemEDE embryonic dorsal epidermisEH embryonic hindgutELDV embryonic/larval dorsal vesselELCS embryonic/larval circulatory systemELO embryonic/larval oenocyteEM embryonic midgutEOLP embryonic optic lobe primordiumEFP external foregut primordiumFA foregut anlageFAISN foregut anlage in statu nascendiFP foregut primordiumGC germ cellHMPP head mesoderm P2 primordiumHMA head mesoderm anlageHA hindgut anlageHPP hindgut proper primordiumIHP inclusive hindgut primordiumLC lateral cordLCG lateral cord glia.LCN lateral cord neuronLVMP longitudinal visceral mesoderm primordiumMA mesectoderm anlageMEP mesectoderm primordiumMIP midline primordiumMAISN mesoderm anlage in statu nascendiMLP midline primordiumMP mesectoderm primordiumNOVNS neuroblasts of ventral nervous systemOSA oenocyte specific anlagePC pole cellPTEA posterior endoderm anlagePTEP posterior endoderm primordiumPMP posterior midgut primordiumPCEA procephalic ectoderm anlagePCEAISN procephalic ectoderm anlage in statu nascendiPCEP procephalic ectoderm primordiumPCN procephalic neuroblastsPEA procephalic ectoderm anlagePEP procephalic ectoderm primordiumPCP protocerebrum primordiumPMP posterior midgut primordiumPP proventriculus primordiumPTEAISN posterior endoderm anlage in statu nascendiSDP salivary duct primordiumSGBSA salivary gland body specific anlageSGDSA salivary gland duct specific anlageSGBP salivary gland body primordiumSNSSA sensory nervous system specific anlageSMP somatic muscle primordiumS subsetTP tracheal primordiumSNSP sensory nervous system primordiumSNSSA sensory nervous system specific anlageTMA trunk mesoderm anlageTMAISN trunk mesoderm anlage in statu nascendiTMP trunk mesoderm primordiumVEA ventral ectoderm anlageVECP ventral ectoderm primordiumVEPP ventral epidermis primordiumVNCP ventral nerve cord primordiumVNA ventral neuroderm anlageVSCSA ventral sensory complex specific anlageVM ventral midlineVMP visceral muscle primordiumVNC ventral nerve cordVP visual primordiumThe authors declare that they have no competing interests.


Adherent cells from carcinomatous pleural effusions of lung cancer patients were tested for their ability to suppress natural killer (NK) cell activity, and the mechanism involved in the suppression of NK cell activity was determined. Adherent effusion cells (AEC) were isolated from malignant pleural effusions of patients by centrifugation discontinuous Ficoll-Hypaque gradients and adherence to serum-coated plastic dishes, and large granular lymphocytes (LGL) were purified from the peripheral blood of normal individuals by centrifugation on discontinuous Percoll gradients and further depletion of high-affinity sheep erythrocyte rosette formation. LGL-mediated lysis of K562 cells was suppressed when LGL were cultured with AEC for 20 h, then washed and tested in a 4-h 51Cr release assay. More profound suppression of NK cell activity was observed when cytotoxicity was assayed in flat-bottomed wells rather than in round-bottomed wells. Cytotoxicity assays conducted at the single cell level in agarose revealed that the frequency of LGL binding to K562 cells and of dead conjugated target cells was reduced after overnight contact with AEC. In agarose microdroplet assays, functional LGL from normal donors exhibited definitive motility, expressing polarized shape. In contrast, a small number of LGL with non-polarized configuration migrated from the agarose droplet after overnight culture with AEC. These results indicate that functionally suppressed NK cells lose their motility, binding capacity and killing activity, which could be responsible for the suppression of NK cell activity by AEC.


Arrhythmogenic right ventricular dysplasia/cardiomyopathy is a disorder characterized by frequent ventricular tachycardia originating from the right ventricle and fibro-fatty replacement of right ventricular myocardium. Though the disorder was originally described during surgical ablation of refractory ventricular tachycardia, catheter ablation of tachycardia is one of the options for patients not responding to anti arrhythmic agents. Direct current fulguration was used in the initial phase followed by radiofrequency catheter ablation. In the present day scenario, all patients with risk for sudden cardiac death should receive an implantable cardioverter defibrillator. Radiofrequency catheter ablation remarkably reduces the frequency of defibrillator therapies. Direct current fulguration can still be considered in cases when radiofrequency ablation fails, though it requires higher expertise, general anesthesia and carries a higher morbidity. Newer mapping techniques have helped in identification of the site of ablation. In general, the success rate of ablation in arrhythmogenic right ventricular dysplasia is less than in other forms of right ventricular tachycardias like right ventricular outflow tract tachycardia. Arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD) is a disorder characterized by fibro-fatty replacement of the right ventricular myocardium, frequent ventricular tachycardia originating from the right ventricle and right heart failure. It was originally described by Fontaine et al during surgical ablation of refractory ventricular tachycardia . The firThough historically the original description of ARVD was during surgical ablation, pharmacological therapy was the initial mode of treatment in most cases. Surgical ablation by right ventricular disconnection was resorted to in resistant cases . PeroperThe initial reports on catheter ablation in ARVD were using direct current fulguration -8. One oRadiofrequency catheter ablation for ARVD has been in use since early nineties . It has Entrainment mapping can be used to characterize reentry circuits in ARVD to guide ablation -15. The Endocardial mapping can detect abnormal fragmented electrograms with delayed potentials. Pacemapping confirms the ablation site by producing a QRS morphology identical to the clinical VT . RecentlThree dimensional Real-time Positioning Management System (RPM) has also been used for guiding ablation in ARVD . RPM useO'Donnell et al have highlighted the electrophysiological differences between patients with ARVD and right ventricular outflow tract tachycardia (RVOT VT) . Though Ablation of ventricular tachycardias in ARVD still remains a clinical challenge, though more and more cases are being reported in the literature -23.


Glutathione S-transferase (GST) isoenzyme expression is altered in a variety of neoplasms and the enzymes are implicated in metabolism of carcinogens and resistance to drugs, including cisplatin. We have studied GST Alpha, Pi, Mu and microsomal isoenzyme expression by immunohistochemistry in normal and cryptorchid testes, intratubal germ cell neoplasia (ITGCN), seminoma and non-seminomatous germ cell tumours. In 16 stage II-IV malignant teratoma intermediate (MTI) both orchidectomy and post-treatment residual surgical masses were studied. All four isoenzymes were strongly expressed in Leydig and Sertoli cells. GST Pi was absent from normal spermatogonia but strongly expressed by the neoplastic germ cells of ITGCN and seminoma. GST Pi was strongly expressed in all elements of teratoma, irrespective of differentiation. There were no qualitative differences in expression between primary and post-chemotherapy metastases. GST Alpha expression in teratoma correlated with epithelial differentiation. GSTs may be important in normal spermatogenesis and protection of germ cells from teratogens and carcinogens. They may have a role in testicular tumour drug resistance but this role is not well defined. GST Pi is a new marker for ITGCN.


Using an indirect immunohistochemical technique on paraffin sections, employing a polyclonal antibody to the acidic  form of glutathione-S-transferase (GST), we have evaluated cytoplasmic and nuclear staining in a series of 67 cervical biopsies including normal non neoplastic tissue, immature squamous metaplasia, all grades of cervical intraepithelial neoplasia (CIN) and invasive carcinomas of keratinising and non-keratinising types. No differences in cytoplasmic staining between the varied lesions studied were seen. However, there were marked differences in nuclear staining. While normal non-neoplastic stratified squamous epithelium showed weak staining of the lower one-third of the epithelium only, in immature squamous metaplasia and in all grades of CIN there was intense nuclear staining in all layers of the epithelium. Invasive carcinomas showed generally less intense nuclear staining than CIN lesions. Endocervical cell nuclei also showed intense nuclear staining. These findings indicate that GST is of limited use as a marker of transformation in the human cervix uteri.