In utero infection of the fetus has become recognized as an important cause of fetal and neonatal morbidity and mortality. Since both anatomic and functional abnormalities have been described in the fetus related to various infections, ultrasonography may be a valuable diagnostic tool in this regard. A complete review of the current literature was undertaken to report available information on this topic. Common pathogens or clinical conditions were selected. The identified data were confounded by the way in which each case originally presented for study. Although certain anomalies were frequently associated with individual organisms, their incidence could not be determined, nor were most specific to that infectious agent. Representative ultrasound images are presented for common and unusual cases.


Ks) in mammalian lineages. However, this positive correlation was observed for lineages with relatively long divergence time such as the human-mouse lineage, and was not found for very short lineages such as the human-chimpanzee lineage. It was previously unclear how to interpret this discrepancy. It may indicate that the positive correlation between ω and Ks in long lineages is a false finding. Alternatively, it may reflect a biologically meaningful difference between various lineages. Finally, the lack of positive correlation in short lineages may be the result of methodological artifacts.We recently reported a highly unexpected positive correlation between the fixation probability of nonsynonymous mutations (estimated by ω) and neutral mutation rate (estimated by Ks as an estimator of neutral mutation rate. Thus, the previously noted lack of positive correlation between ω and Ks in short lineages is due to stochastic noise in Ks that makes it a far less reliable estimator of neutral mutation rate in short lineages as compared to long lineages.Here we show that a strong positive correlation can indeed be seen in short lineages when a method was introduced to correct for the inherently high levels of stochastic noise in the use of Ks can be observed in all mammalian lineages for which large amounts of sequence data are available, including very short lineages. It confirms the authenticity of this highly unexpected correlation, and argues that the correction likely applies broadly across all mammals and perhaps even non-mammalian species.A positive correlation between ω and  Ks (or Ka). Ks is typically treated as a reasonable proxy for neutral mutation rate, and the Ka/Ks ratio (abbreviated ω) is often used as a measure for the fixation probability of nonsynonymous mutations scaled to neutral expectation.Point mutations in coding regions of genes can be classified into two categories: synonymous and nonsynonymous. In mammals, synonymous mutations are largely neutral, though they may sometimes experience weak selection . NonsynoKs in several mammalian lineages, arguing that the fixation probability of nonsynonymous mutations in a gene is positively corrected with the gene's neutral mutation rate [Ks, and in fact, simulations demonstrate that only an inverse correlation should in theory exist between ω and Ks due to a mathematical artifact [Ks therefore challenges the current paradigm of molecular evolution, and necessitates a reexamination of the widely held assumption that ω is determined primarily by selective pressure independent of neutral mutation rate.Recently, we discovered a strong positive correlation between ω and ion rate . This fiion rate -5. Conveartifact  , but not found in the human-chimpanzee or the human-macaque lineages, which have much lower levels of sequence divergence . Instead, an inverse correlation between ω and Ks was seen in these latter short lineages [Ks observed in long lineages may actually be an artifact associated with the imprecise calculation of Ka or Ks. In particular, multiple hits at any given nucleotide position can occur with high probability in long lineages. Such multiple hits, difficult to correct since only one or zero changes can be observed at a given nucleotide position, can constitute a major source of error in the calculation of Ka and, even more so, Ks. It is formally possible, therefore, that errors stemming from the improper correction of multiple hits along with some other unknown factors have systematically biased the calculation of Ka and Ks in long lineages, in a manner that creates an artifactual positive correlation between ω and Ks.There is, however, an important caveat to this finding. The positive correlation between ω and lineages   represents a much greater fraction of μ in short lineages as compared to long lineages, and our simulations showed that this could indeed lead to a strong inverse correlation between ω and Ks in very short lineages such as the human-chimpanzee lineage because Ks is also used as the denominator in the calculation of ω  .Ks in short lineages is indeed due to the inherently high levels of stochastic noise affecting Ks in short lineages, then a reasonable correction of noise in Ks might be able to bring out the positive correlation again. Here, we show that by introducing a method that corrects for stochastic noise in Ks, a strong positive correlation between ω and Ks indeed becomes observable in short lineages. This result strengthens the authenticity of this highly unexpected correlation, and argues that a biological mechanism  is relating the fixation probability of nonsynonymous mutations to mutation rate.If the failure to observe a positive correlation between ω and Ks in human-mouse and mouse-rat comparisons. Indeed, for both lineages, a highly robust correlation exists and binning of genes creates a visually striking representation of the correlation , the relationship between ω and Ks goes from a strong positive correction in the case of long lineages progressively toward a strong inverse correlation in the case of very short lineages.We then considered the human-chimpanzee and human-macaque ortholog pairs within the complete data set. As expected for these short lineages, plotting ω against lineages . If thisKs is frequently used as a proxy for neutral mutation rate (μ). It is often forgotten, however, that mutational events are a discreet process and as such are subject to stochastic variation. Over short periods of evolutionary time, this stochastic variation is often large relative to the true underlying neutral mutation rate. As evolutionary time lengthens and the number of mutational events increases, stochastic variation decreases relative to neutral mutation rate. This is clearly demonstrated by computer simulations in our previous study [In molecular evolutionary studies, us study .Ks can lead to an artifactual inverse correlation is because Ks is in both parameters being corrected, and it is the denominator of ω. Mathematically, by just correcting for noise on the x-axis (Ks), the artifactual inverse correlation should go away even if there is still considerable noise on the y-axis. This prediction was clearly borne out by our previous simulation studies [Ks only, and to examine if such correction can eliminate the artifactual inverse correlation between ω and Ks in short lineages. This may reveal the underlying positive correlation, if any, that has been obscured.High levels of stochastic noise in  studies . We therKs value from orthologs of the same gene but in a longer lineage . Indeed, when we plotted ω of the human-chimpanzee lineage against Ks of the corresponding human-mouse lineage, not only did the inverse correlation disappear, but a positive correlation as typically observed in long lineages was seen . This may introduce confounding effects. We therefore also plotted human-chimp or human-macaque ω against mouse-dog Ks. We found that a rather similar positive correlation between ω and Ks exists despite a complete lack of shared descent between the lineage used to calculate ω and the lineage used to calculate Ks  than to use the observed Ks from the short lineage. This study and our unpublished data confirm that neutral mutation rate can remain relatively stable across different mammalian lineages [Another important message from the study is that lineages ,9.Ks and the effect of polymorphisms are not major issues. In the study of short lineages, the converse situation is true. It is thus gratifying that a strong positive correlation between ω and Ks can be observed not only for long lineages, but also for short lineages after noise in Ks is corrected. Together, these results argue that the correlation is the result of a biological mechanism rather than a methodological artifact.Neither the previous long-lineage study nor the current short-lineage study alone is sufficient to eliminate methodological artifacts from consideration. In the study of long lineages, the correlation is complicated by possible difficulties in correcting for multiple hits, though the stochastic noise in Ks. Neither this study nor our previous study provides a definitive mechanism by which this correlation may occur, though several models have been proposed. Perhaps ω is reflective of some combined effect of selective pressure and neutral mutation rate, or perhaps selective pressure and neutral mutation rate influence each other in some way [It is as yet unclear why there should exist such a strong positive correlation between ω and some way . It was some way -14, may some way . It was some way , that issome way -17. ThesGiven the presence of the correlation in all the mammalian lineages for which there is sufficient genome sequence data, including very short lineages, it now seems clear that the correlation is indeed the result of a biologically meaningful process. The stage is set for future studies to identify the mechanisms underpinning this enigmatic correlation.Homo sapiens (NCBI 35); chimpanzee, Pan troglodytes (PanTro 1.0); rhesus macaque, Macaca mulatta (Mmul 1.0); mouse, Mus musculus (NCBI m34); rat, Rattus norvegicus (RGSC 3.4); and dog, Canis familiaris (CanFam 1.0). Genes were clustered into orthologous groups using reciprocal best BLAST hits following established methods [Ks cut-offs at roughly three standard deviations above average for all alignments. Sequences were curated for length differences and for poor alignment. This resulted in 5,831 orthologous groups containing a member from each of the six species with reasonable alignment and 6,779 orthologous groups containing all five primate and rodent species. Ortholog groups, alignments, and evolutionary estimates used in this study can be obtained through the SPEED database [Using data from Ensembl v36 , sequenc methods -21. We f. Evolutionary parameters were estimated using the Li method [Ks) when other methods such as PAML [In-frame alignments of orthologs were performed using The Wisconsin Package v10.2 i method , though  as PAML ,24 were The author(s) declare that there are no competing interests.EJV and BTL designed the project, performed analysis and wrote the paper. EJV performed bioinformatic data mining. Both authors have read and approved the final manuscript.Ks for the human-mouse lineage and the mouse-rat lineage.Supplementary Figure 1. Positive correlation between ω and Click here for fileKs in the human-chimpanzee lineage and the human-macaque lineages.Supplementary Figure 2. Lack of positive correlation between ω and Click here for fileKs or K4 between different lineages.Supplementary Figure 3. Correlation of Click here for file


At least two downfield shifted and broad paramagnetic signals were apparent in the 1H NMRspectrum. Temperature dependence of chemical shift was exploited in order to estimate the diamagnetic shiftof the signals in the diamagnetic region of the spectrum. 2D TOCSY, NOESY, COSY and 1H-3C HMQCspectra revealed the presence of aromatic protons (which were assigned as His residues), Gly and some othershort chain amino-acids. Combinations of the molecular masses of such components together with acetate(which is present in the solution) and chromium atoms allowed a tentative proposal of a model for thecompound.A low-molecular weight chromium-containing fraction of the material resulting from dichromatereduction by bovine liver homogenate was investigated by NMR and ES-MS. The ES-MS spectrum showed areadily detectable peak at m/z = 786.1. The same molecular weight reasonably agreed with the relatively lowdiffusion coefficient measured by NMR-DOSY experiments on the main species observed in the


The IPDC (http://nihroadmap.nih.gov/molecularlibraries/ipdc/) is dedicated to the production of known and novel molecular imaging probes, with its services currently being used by the NIH intramural community, although in the future it is intended that the extramural community will also benefit from the IPDC’s resources. The Center has been set up with the belief that molecular imaging, and the probe chemistry that underpins it, will constitute key technologies going forward. As part of the larger molecular libraries and imaging initiative, it is planned that the IPDC will work closely with scientists from the molecular libraries effort. Probes produced at the IPDC include optical, radionuclide and magnetic resonance agents and may encompass any type of contrast agent. As IPDC is a trans-NIH resource it can serve each of the 27 Institutes and Centers that comprise NIH so its influence can be expected to impact widely different subjects and disease conditions spanning biological research. IPDC is expected to play a key part in interdisciplinary collaborative imaging projects and to support translational R&D from basic research through clinical development, for all of the imaging modalities. Examples of probes already prepared or under preparation are outlined to illustrate the breadth of the chemistries undertaken together with a reference outline of the diverse biological applications for which the various probes are intended.The Imaging Probe Development Center (IPDC), part of the NIH Roadmap for Medical Research Initiative ( One of three major themes was identified as “New Pathways to Discovery” one of which, in turn, comprised the Molecular Libraries and Imaging Initiative (http://nihroadmap.nih.gov/molecularlibraries/). The Libraries Roadmap Initiatives have put in place personnel and facilities which enable the large-scale screening of hundreds of thousands of small molecules as molecular probes which can be used to study the full spectrum of cellular and biochemical processes [In late 2004 the US National Institutes of Health launched its Roadmap for Medical Research in the 21rocesses . This aphttp:// nihroadmap.nih.gov/molecularlibraries/ipdc/). The field of molecular imaging has also been undergoing profound changes in the last few years spurred by advances in detection capabilities in the major modalities of magnetic resonance imaging, radionuclide imaging using both single photon and positron emitting nuclides, and most notably, in fluorescence microscopy [Concomitant with, and complementary to, the Libraries Initiatives are the Roadmap Molecular Imaging Initiatives which include an imaging agent database (MICAD), an extramural research effort aimed to dramatically increase the sensitivity and specificity of molecular imaging agents, and a core chemistry facility to actually synthesize imaging probes – the Imaging Probe Development Center (IPDC) . It is clearly of vital importance that low molecular weight binding entities identified from screens and optimized for binding via analog syntheses are then only modified via chemical handles in positions that have minimal adverse effects on their target binding capabilities as well as on the detectable agent’s properties.The goal of the IPDC, therefore, is to provide a mechanism to produce “known” or published agents for which there is no commercial supply, and to generate novel imaging probes, for biomedical research and clinical applications. The IPDC is composed of two major elements, (a) receptor modeling/biophysical chemistry, and (b) organic synthesis and it is intended that IPDC will complement and support the screening centers set up under the Libraries initiative since molecular imaging can be seen as a logical downstream application of the screening centers discovery efforts. We began operations after moving into a newly refurbished facility near the end of 2006, and initially solicited proposals for new imaging probe compositions from the NIH intramural community. At the same time we recruited a staff of young scientists with diverse interests and expertise related to imaging agents and centered on one or more of classical synthetic, polymer, bioconjugate, metal complexation chemistries or radiochemistry. Along with the new staff, multiple new pieces of equipment {Flash, HPLC, FPLC, MS – ES,-CI, -MALDI, NMRs, etc.} were obtained, installed and trained on for the simultaneous preparation of any type of composition we might be asked to produce in the coming years. In addition to the chemistry production, IPDC has biological evaluation facilities which will enable it to perform preliminary Agents used for molecular imaging at this point in time can run the full spectrum from low molecular weight drug-like molecules through natural products, peptides, nucleotides, sugars and their oligo- and combined conjugates, polymeric, proteinaceous and nanoparticle-like moieties, and more; all tagged with one or more of MRI, radionuclidic, ultrasound or fluorescently detectable labels.1 and represents a broad array of interests both with regard to human conditions/diseases and areas of basic science studies. All proposals are discussed in detail at monthly steering committee meetings prior to any acceptance or commitment on the part of the IPDC. Proposals submitted to the IPDC from the intramural NIH community must first be approved for submission by the Scientific Director of the Institute wherein the submitting scientist is located. In addition, IPDC is overseen by the Roadmap’s Office and Portfolio Analysis and Strategic Planning (OPASI), which oversees the entire Roadmap program, and monitors the IPDC through the Molecular Libraries and Imaging Implementation Group (MLIIG). The MLIIG, in turn and as the title implies, oversees the molecular libraries and imaging programs. Under these umbrellas IPDC sent out three solicitations for new project proposals from the NIH intramural community during its first year or so of existence.IPDC operates under the direction of a steering committee with members drawn from chemists and imaging specialists representing most of the Institutes with an interest in imaging and/or chemistry. The current make up of this committee in shown in Table 1), with the largest number of projects undertaken on behalf of the National Cancer Institute (NCI). Probe requests have covered the full spectra of chemical compositions used for molecular targeting from small molecular weight compounds from medicinal and natural products chemists through peptides, nucleotides, sugars and their analogs and oligomers, to compositions based on proteins, antibodies and their constructs, dendrimers, nanoparticles and liposomes. The detectable agent most requested for compositions is based on fluorescence as an imaging beacon, although a significant fraction is related to metal complexes for MRI studies and radionuclide-based agents.Very strong interest and a very strong response throughout our first year led to the adoption of several dozen projects for the supply of imaging probes to multiple Institutes and Centers within NIH . Over th2 lists a selection of the probes made or under preparation by composition, detectable agent, the major requested indication, and NIH Institute. Known fluorescent analogs such as biarsenicals used for site-specific tetracysteinyl-protein tagging, which are tricky to make even for a well-trained chemist, have now been requested by several groups so that around half-a- dozen analogs have been produced. Requested originally for prion protein infection studies, they clearly can be applied as a research tool for any indication using a tetracysteinyl-labeling strategy. Fluorescent dyes are generally quite difficult to work with on a chemical basis as such preparations are generally more impure than most chemical starting materials, with confounding mixtures often involving several similar compounds as well as a selection of isomeric variants, none of which is usually of importance to a biological stain- or assay-minded biologist. Many fluorescent dyes are therefore repurified routinely prior to attempting further chemical manipulations.Table Also widely requested are targeting agents using common dyes such as the Alexa and cyanine series, for instance cholesterol-linked dyes for lipid raft studies and GABA analogs for calcium channel neurotransmitters. Others are involved in basic studies designed to improve dye properties such as by modifying fluorescent lifetimes. Particularly interesting probes seek to dramatically improve fluorescent imaging by applying ‘caged’ agents, that is, agents that are non-fluorescent until an appropriate light source renders them so. Such agents are designed to make a dramatic leap forward in fluorescent imaging technology by increasing sensitivity toward tracking single molecules using photoactivation localization microscopy (PALM). PALM may allow researchers to break the diffraction barrier limit by selectively activating and detecting one molecule at a time, repeating the procedure numerous times, and then reconstructing a whole image from the accrued data. A different approach seeks to make a similar leap toward ‘super-resolution’ by using a stimulated emission depletion-like (STED) strategy employing Förster resonance energy transfer (FRET) pairs. The activatable FRET donor-acceptor technique is also applied in several other studies, for instance to probe HIV-1 infection mechanisms. This may have general implications for all diseases involving similar viral replication pathways where it greatly improves the target-to-background ratio. A separate and important series of caged probes are non-fluorescent caged glutamates which are used in receptor activation studies.Intracellular event studies, outside of infection, are also well represented among our probe requests with enzyme substrates, for example, with a phosphatidylinositol-specific phospholipase C (PI-PLC) already being made. These complex and non-fluorescent multi-phosphatidylinositols will become fluorescent upon PLC cleavage and will offer a unique way to look at this important class of enzymes. Separately, we are working on specifically designed species to target intracellular compartments such as lysosomes and such agents can have an impact on all lysosomal storage diseases. This project was undertaken with the NCGC , who successfully identified targeting moieties of interest, and it represents the first example of these two new Roadmap initiatives working together on a common problem along with the requesting Institute.Some fluorescent molecules may never be more than highly specialized research tools due to their being unavailable merely by reason of the difficulties involved in their syntheses. However, such agents may also offer special insights into basic cellular processes. Such a series of probes is illustrated by the pteridine nucleic acid analogs which are fluorescent analogs of the guanosine and adenosine nucleosides. Although very difficult to make, with up to 15 or more synthetic steps involved, often on highly labile intermediates, their unique capacity to be incorporated directly into the oligonucleotide chains of DNA and RNA could offer unique perspectives on basic oligonucleotide studies. They may even have future roles to play in studying the ‘small’ and ‘micro’ RNAs which are of so much current interest.in vivo studies, although some of the fluorescent agents mentioned can also be applied to animal imaging. The nitroxides are redox sensitive probes and are envisaged as agents for detecting the effects of radiation therapy. They are being tested in the context of radiotherapy for glioma. This agent potentially has broad application in monitoring radiation therapy. The manganese complexes are intended as bone imaging agents, as their chelate structures have appended phosphate groups, while it is also an interesting challenge from a chemist’s point-of-view as to whether a truly stable manganese-chelate complex for in vivo use can be obtained. The more common gadolinium ion has been attached to series of dendrimers using stable chelates for several applications including vascular and lymphatic imaging and as a new set of agents which may be able to traverse blood-brain-tumor barriers. Low molecular weight gadolinium-labeled agents are being studied as neural tracers to map connections within the brain.With MRI reagents the IPDC has been asked to prepare low and high molecular weight gadolinium complexes, manganese complexes and nitroxide radicals. These are generally used for ® molecules, which are small enough to be chemically synthesized on a peptide synthesizer and also large enough to be produced from biotechnological cell fermentations. These picomolar affinity constructs represent a new platform for disease targeting, can be tagged with detectable agents from any of the modalities, and are being developed for both therapy and imaging uses.Radiolabeled species represent the majority of animal and particularly human-use imaging probes. The IPDC is working on low molecular weight radiolabeled thyroid hormones for PET imaging of hypothyroid conditions, radiolabeled adenosine analogs for imaging adenosine receptors, radiolabeled dopamine receptor antagonists for dopamine receptor imaging, and radiolabeled vasopressin analogs for CNS receptor imaging. Proteins are also under study with serum amyloid and ‘Frizzled’ proteins being radiolabeled while ongoing radiolabeling work continues with antibodies. One satisfying aspect of the IPDC’s scope is that imaging sciences, more than most, bridges biotechnology and chemistry, and even includes nanotechnology as we are now producing nanoparticle-derived agents and are likely to do more of such work in the future. On a more compositional note we have also prepared targeting liposomes for use in studies targeting colon cancer. A notable example of our bridging, interdisciplinary position is our ongoing work with AffibodyMolecular imaging is becoming ever more important in a broad range of studies being done from the smallest intracellular level through intact organisms, including humans. In addition, molecular imaging is being adopted by drug makers and biopharmaceutical firms as a useful adjunct to development work, diagnoses, therapy decision-making, and disease monitoring. Indeed, one might contend that many drugs of the future will need to use some kind of imaging work to justify their application, and as molecular imaging becomes ever more exact and sensitive that position can only be reinforced. It is intended that the IPDC will have the capacity to assist in the drug development process and in the transfer of new agents into initial clinical trials. Our capacity to make almost any reasonable imaging probe composition should make IPDC a useful resource, especially for scientists who may have no current chemistry experience or available collaborating chemists and may be blocked from carrying out what may be otherwise important investigations. But, as has also been demonstrated with our initial projects, even medicinal chemists can make use of the IPDC to do jobs that may be too difficult in their own laboratory, even if from just a regulatory viewpoint, as for instance with radiolabeling. The plethora of new agents being described in the literature clearly leaves many bioscientists frustrated that studies cannot be repeated and expanded in new directions often due to lack of chemistry resources, and the IPDC will seek to address this issue to the best of its abilities. griffithsgl@mail.nih.gov.The IPDC looks forward to addressing the needs of the broader imaging community. The initial experience bodes well for the ability of IPDC to provide unique imaging probes that will provide biologic insights into the mechanisms of a broad spectrum of disease. Meanwhile I encourage those of you who are interested to feel free to contact me if you have questions about the IPDC or its services, or to enquire informally about a prospective molecular imaging probe that you may be interested in. I can be reached by email at:


At least two downfield shifted and broad paramagnetic signals were apparent in the 1H NMRspectrum. Temperature dependence of chemical shift was exploited in order to estimate the diamagnetic shiftof the signals in the diamagnetic region of the spectrum. 2D TOCSY, NOESY, COSY and 1H-13C HMQCspectra revealed the presence of aromatic protons (which were assigned as His residues), Gly and some othershort chain amino-acids. Combinations of the molecular masses of such components together with acetate(which is present in the solution) and chromium atoms allowed a tentative proposal of a model for thecompound.A low-molecular weight chromium-containing fraction of the material resulting from dichromatereduction by bovine liver homogenate was investigated by NMR and ES-MS. The ES-MS spectrum showed areadily detectable peak at m/z 786.1. The same molecular weight reasonably agreed with the relatively lowdiffusion coefficient measured by NMR-DOSY experiments on the main species observed in the


To evaluate whether some form of mild immunosuppression may influence the geographical distribution of non-AIDS Kaposi's sarcoma (KS), we correlated incidence rates of KS and non-Hodgkin's lymphoma in individuals aged 60 or more in 18 European countries and Israel. Significant positive correlations emerged but, within highest risk countries , internal correlations were inconsistent. © 1999 Cancer Research Campaign


Both AMPA- and NMDA-mediated components were suppressed, and paired-pulse facilitation was also enhanced indicating that the suppression is mediated largely by reduced glutamate release. Blockade of D2-like receptors greatly reduced the suppression of EPSPs. Dopamine also lowered input resistance, and reduced the number of action potentials evoked by depolarizing current steps. The drop in input resistance was mediated by activation of D1-like receptors, and was prevented by blocking K+ channels with TEA. The dopaminergic suppression of synaptic transmission is therefore mediated by a D2 receptor-dependent reduction in transmitter release, and a D1 receptor-dependent increase in a K+ conductance. This suppression of EPSPs may dampen the strength of sensory inputs during periods of elevated mesocortical dopamine activity.Dopaminergic projections to the superficial layers of the lateral entorhinal cortex can modulate the strength of olfactory inputs to the region. We have found that low concentrations of dopamine facilitate field EPSPs in the entorhinal cortex, and that higher concentrations of dopamine suppress synaptic responses. Here, we have used whole-cell current clamp recordings from layer II neurons to determine the mechanisms of the suppression. Dopamine (10 to 50  The entorhinal cortex is animportant interface that links primary sensory and association cortices to thehippocampal formation, and it is critical for the sensory and mnemonicfunctions of the medial temporal lobe –4. In th1 receptors cansuppress glutamate release in layer V [1 receptor activation on working memory follows an inverted U-shaped function[1 receptors can also haveopposite effects on NMDA receptor-mediated synaptic currents [Ih current [The large dopaminergic projectionto the prefrontal cortex is known to regulate cellular processes related toworking memory –21, and layer V –24 but c layer V , 26. Furfunction, and strcurrents , 28. We currents . Higher currents  and layecurrents . Dopamincurrents  and redu current . Dopamin2-likereceptor-mediated suppression of transmitter release, we show evidence thatEPSPs are also reduced by an increased K+ conductance dependent onactivation of D1 receptors. We have used whole-cell currentclamp recordings to investigate the mechanisms of the suppression of EPSPs bydopamine in electrophysiologically identified “fan” cells in layer II of thelateral entorhinal cortex. Receptorblockers were used to determine the dopamine receptors that mediate thesuppression of EPSPs, and paired-pulse tests were used to assess whether thesuppression is expressed pre- or postsynaptically. Changes in the intrinsic excitability of fancells were also monitored using responses to hyperpolarizing and depolarizingcurrent steps. In addition to a D2 and 5% CO2 containing (in mM) 124 NaCl, 5KCl, 1.25 NaH2PO4, 2 MgSO4, 2 CaCl2,26 NaHCO3, and 10 dextrose . All chemicals were obtained from Sigma-Aldrich, Mo, USA. Horizontal slices (300 μm thick) were cutusing a vibratome ,and slices recovered for at least one hour at 22 to 24°C. Slices were transferred individually to arecording chamber and visualized using an upright microscope equipped with differential interference contrast optics, a 40x water immersionobjective, and a near-infrared camera . Submerged slices were superfused withoxygenated ACSF at a rate of 1.5 to 2.0 mL/min. Slices containing the lateral entorhinal cortex were taken from ventralsections about 1.9 to 3.4 mm above the interaural line [μm from the cortical surface [Methods for obtaining whole cell current clamp recordingswere similar to those described previously , 33, 34.ral line . Layer I surface –39.2, 10 HEPES, 0.5 EGTA, 2 ATP-Tris, and 0.4 GTP-Tris. Pipettes were placed in contact with somataof layer II neurons, and gentle suction was applied under voltage clamp to forma tight seal (1–3 GΩ). Whole cell configuration was achieved byincreased suction, and experiments began after cells stabilized . Current clamp recordings were obtained using an Axopatch 200B amplifierand displayed on a digital oscilloscope (Gould 1604). Recordings were filtered at 10 kHz anddigitized at 20 kHz  for storage on computer harddisk. Recordings were accepted if theseries resistance was ≤25 MΩ (mean = 16.9 ± 0.9 MΩ) and if input resistance andresting potential were stable. A bipolarstimulating electrode made from two tungsten electrodes  waspositioned to span layer I near the border with layer II approximately 0.2 to0.6 mm rostral to the recording electrode. Synaptic responses were evoked with 0.1 millisecond constant currentpulses delivered using a stimulus timer and isolation unit . Stimulation intensity wasadjusted to evoke responses approximately 75% of maximal (75 to 300 μA).Patch recording pipettes for whole cell recordings wereprepared from borosilicate glass  using a horizontalpuller  and were filled with a solution containing (in mM) 140 K-gluconate,5 NaCl, 2 MgCln = 118) included for analyses were identified as “fan” cells based onelectrophysiological characteristics described previously [All neurons was coapplied toslow the oxidation of dopamine [The effects of dopamine on glutamate-mediated synaptictransmission in the lateral entorhinal cortex are largely uncharacterized. We therefore recorded both mixed and isolatedcomponents of excitatory postsynaptic potentials (EPSPs) evoked by stimulationof layer I before and after 5-minute bath-application of 1, 10, or 50 torhinal –32 andptorhinal , 43 cortdopamine , 31, 43,μM dopamine. Stimulation intensity was reduced to evokeEPSPs approximately 50% of maximal, and ten responses were averaged foranalyses. Paired-pulse facilitation wasquantified by expressing the amplitude of the second response as a percentageof the first response.Paired-pulsetests were used to determine whether dopamine modulates EPSPs through a pre- orpostsynaptic mechanism . Pairs oμM dopamine on isolated components of synapticresponses. After baseline recordings innormal ACSF, AMPA receptor-mediated responses were isolated with bathapplication of 50 μM 2-amino-5-phosphonovaleric acid (APV) and 25 μMbicuculline methiodide, or NMDA receptor-mediated responses were isolated with20 μM 7-nitro-2,3-dioxo-1,4-dihydroquinoxaline-6-carbonitrile (CNQX) and 25 μMbicuculline. GABA-mediated IPSPs wereisolated with either 1 mM kynurenic acid or 20 μM CNQX with 50 μM APV. Isolated synaptic responses were recordedbefore and after 5-minute application of 50 μM dopamine. Isolated AMPA receptor-mediated responseswere also used to determine if dopamine suppresses EPSPs primarily through D1-or D2-like receptors. Baseline responses were recorded in the presence of either the D1 receptor antagonist SCH23390 (50 μM) or the D2 receptor antagonistsulpiride (50 μM) [μM dopamine was then applied for 5 minutes. Sulpiride was prepared daily in a stocksolution of 6% DMSO in ACSF titrated with 0.1 N HCl, and there was a finalconcentration of 0.1% DMSO with sulpiride.Mechanisms mediating thesuppression of EPSPs by high concentrations of dopamine were investigated byassessing the effects of 50  (50 μM) –31, and μM dopamine. The number of action potentials elicited in response to suprathresholdcurrent injection can be used to characterize neuronal excitability [μM tetrodotoxin (TTX)or 30 mM tetraethylammonium (TEA). Blockers were preapplied for 5–10 minutes priorto coapplication of dopamine for 5 minutes.The effects of dopamine on theintrinsic excitability of fan cells were assessed by monitoring responses tohyperpolarizing and depolarizing current steps. Changes in action potentials, afterhyperpolarizations, input resistanceand inward rectification were examined before and after 5-minute bathapplication of 1, 10, or 50 tability , andwe tability . Receptot-tests or mixed design ANOVAs.Electrophysiologicalcharacteristics of fan cells and changes in synaptic responses were analyzedusing the software program Clampfit 8.2 . The amplitudes of averaged EPSPs weremeasured relative to the prestimulus baseline, and paired-pulse facilitationwas determined by expressing the amplitude of the second response as aproportion of the amplitude of the first response. Action potential amplitude was measured fromresting potential, and action potential width and fast and mediumafterhyperpolarizations were measured from threshold. Input resistance was calculated by measuringpeak and steady-state voltage responses to −200 pA currentsteps (500 milliseconds), and inward rectification was quantified by expressingthe peak input resistance as a proportion of the steady-state resistance(rectification ratio). All data wereexpressed as the mean ±SEM for plotting, and changes in response propertieswere assessed using paired samples A total of 118 fan cells in layerII of the lateral entorhinal cortex were identified electrophysiologically andincluded for analysis, and the characteristics of these cells were similar tothose reported previously , 41. FanμM dopaminefacilitated fEPSPs and 50 to 100 μM dopamine suppressed fEPSPs [μM dopamine resulted in a strong suppression of synaptic response to 38.5 ± 5.8% of baseline levels  that could be reversed by 15 minutes washout in normal ACSF (3cells). We initially expected 10 μMdopamine to facilitate EPSPs [μM dopamine instead causeda small synaptic suppression . However, a lower concentration of 1 μM dopamine significantly enhancedresponses to 132.7 ± 4.4% of baseline levels . In our previous study using a gas-fluid interface chamber, a larger bathvolume and slower flow-rate may have increased dopamine oxidation and reducedthe effective concentration of dopamine at the slice, and this may account forwhy a higher applied concentration facilitated responses in that study[n = 8).We previously foundconcentration-dependent effects of dopamine on field EPSPs in layer II in vitro, in which 10 d fEPSPs . We obtate EPSPs , but fouat study. Bathapt13 = 2.78, P < .05 for 10 μM; t8 = 2.97, P < .05 for 50 μM), suggesting that dopamine reduced EPSPs bysuppressing glutamate release. Incontrast, the low concentration of 1 μM dopamine that facilitated EPSPs had nosignificant effect on paired pulse facilitation  and the AMPA component was reduced to 41.7 ± 5.6% of baseline . The suppression of EPSPs by high concentrations of dopaminewas examined more closely using pharmacologically isolated synapticresponses. Consistent with a suppressionof glutamate release from presynaptic terminals, bath application of 50 μM dopamine in the presence of either the D1 receptor antagonist SCH23390 (50 μM) or the D2 receptor antagonistsulpiride (50 μM). Similar to previousreports that have used selective agonists in the medial [1 agonistSKF38393  or the D2 agonist quinpirole  had no effect on EPSPs (data not shown), and we therefore usedreceptor blockers known to affect synaptic responses in the lateral entorhinalcortex [1 antagonist SCH23390 did notblock the suppression of AMPA-mediated EPSPs , suggesting that D1 receptors do notmediate the suppression. However, blockade of D2 receptors withsulpiride significantly reduced the effects of dopamine on AMPA-mediatedEPSPs. Coapplication of dopamine withsulpiride (n = 5) resulted in a nonsignificant suppression of synapticresponses, and the size of the suppression was significantly smaller than thatobserved with dopamine alone , suggestingthat a non-D2 receptor-mediated mechanism mediates the residualsuppression.Dopamine receptor subtypesunderlying the suppression of AMPA-mediated synaptic responses wereinvestigated by applying 50 e medial , 31 and e medial  entorhinlcortex . ApplicaμM APV and 20 μMCNQX to block ionotropic glutamate transmission. A concentration of 50 μM dopamine suppressedboth the early GABAA- and late GABAB-mediated componentsof the IPSP. The early IPSP was reducedto 84.5 ± 8.7% of baseline levels, and the late IPSP was reduced to 62.3 ± 11.1%of baseline levels . The dopaminergic suppression of GABA synapsesindicates that the reduction of EPSPs by dopamine is unlikely to be due toincreased GABAergic inhibition of fan cells.Biphasic IPSPs were recorded fromfan cells held near action potential threshold (−51 to −48 mV) andexposed to either 1 mM kynurenic acid or a combination of 50 t8 = 4.73, P < .001; n = 9), and peak inputresistance was reduced from 90.3 ± 7.6 to 68.9 ± 3.1 MΩ by 50 μM dopamine . Similar changes in membrane potential andinput resistance were observed for 10 μM dopamine (not shown) and have alsobeen reported following application of high concentrations of dopamine inwhole-cell recordings from medial entorhinal cortex stellate cells [μM dopamine did not show a drop in input resistanceor hyperpolarization of membrane potential.Bath application of dopamine alsohyperpolarized resting membrane potential and reduced the input resistance offan cells. Membrane potential wasincreased from −56.1 ± 2.0 to −59.7 ± 1.4 mV .In layer V entorhinal cortexcells dopamine causes a reduction in excitability and a drop in inputresistance through an increase in the hyperpolarization-activated current Ih , and chaμM dopamine reduced thenumber of action potentials evoked by brief 500 milliseconds depolarizingcurrent pulses . A higher 50 μM concentration of dopamine caused a similar reduction in the number of spikes(from 3.9 ± 0.2 to 2.8 ± 0.6) that was not statistically significant . Thereduction in spiking could result in part from reduced input resistance, but itwas not due to membrane hyperpolarization because cells were tested at the samemembrane potential both before and after dopamine application.Dopamine suppressed theexcitability of fan cells, and application of 10 and 50 lses see . ThenumμM dopamine was blocked by coapplication of theD1 receptor antagonist SCH23390 . It hasbeen suggested that the reduced input resistance induced by dopamine in medialentorhinal cortex stellate cells might be mediated by an increased K+ conductance [+ channel blocker TEA . Coapplication of TEA blocked thereduction in input resistance induced by dopamine may be responsible for the drop in input resistance observed here. Background leak channels are insensitive toTEA [+ currents areblocked by TEA, but dopamine in the prefrontal cortex tends to enhance neuronalexcitability by suppressing thesecurrents . In the prefrontalcortex, D1 receptor activation typically inhibits IRKC by direct effects of cAMP on IRK channels, but strongactivation can increase IRKC via phosphorylation of the channels throughelevated levels of PKA [1 receptor-dependent K+ conductance in fancells.A large number of Ke toTEA  and are see also , 63). Seurrents s of PKA . This co1 receptor antagonist SCH23390 prevented the dopamine-induced reduction in inputresistance it did not completely eliminate the reduction in the number ofspikes, suggesting that reduced input resistance cannot entirely account forthe reduction in spiking, and that other mechanisms may also contribute. D1 receptor activation can increase spiking in prefrontal neuronsby enhancing the persistent Na+ current (INaP) and suppressing a slowly-inactivating K+ conductance [INaP has also been observed[Ih [Ih in fan cells, andaction potential threshold and afterhyperpolarizations were not affected,suggesting that the underlying currents were not modified. Dopaminergic effects on INaP were not directly assessed in the present study,and the drop in input resistance could mask possible reductions in depolarizingresponses to current injection related to INaP. However, in tests in which SCH23390 preventeda change in input resistance, we found no reduction in the response to +20 pApulses. This argues against a D1-mediatedreduction in INaP, but itis still possible that dopamine may reduce spiking via a D2 receptor-mediated reduction in INaP [ We observed a decrease in fancell firing during depolarizing current steps after dopamine, and the reducedspiking may reflect the drop in cellular input resistance. A surprising finding was that while the Dductance , 70, butobserved. In layeved[Ih . Here, tin INaP . We have shown here that dopaminehas concentration-dependent, bidirectional effects on glutamate-mediatedsynaptic transmission in principal cells of layer II of the lateral entorhinalcortex. The lateral entorhinal cortex receivesa major input from the piriform cortex –7, and d


Amine neurotransmitters, such                        as dopamine, serotonin, and noradrenaline, play important roles in the                        modulation of behaviors and metabolism of animals. InC. elegans, it                        has been shown that serotonin and octopamine, an invertebrate equivalent of                        noradrenaline, also regulate lifespan through a mechanism related to food                        deprivation-mediated lifespan extension. We have shown recently that                        dopamine signaling, activated by the tactile perception of food, suppresses                        octopamine signaling and that the cessation of dopamine signaling in the                        absence of food leads to activation of octopamine signaling. Here, we                        discuss the apparent conservation of neural and molecular mechanisms for                        dopamine regulation of octopamine/noradrenaline signaling and a possible role                        for dopamine in lifespan regulation.                 Drosophila, it is shown that a quantitative                            trait locus for the variation of longevity maps into the aromatic L-amino acid                            decarboxylase gene, which is required for dopamine and serotonin synthesis [C. elegans, the serotonin receptor                            mutant ser-1 has increased lifespan whereas another serotonin receptor                            mutant ser-4 has decreased life span, suggesting that serotonin can                            affect lifespan in opposite ways depending on the receptor mechanism that is                            invoked [ It is becoming clear from                            studies in model animals that amine neurotransmitters can regulate the                            longevity of animals. In ynthesis .        invoked . It is a invoked .       C. elegans                         [C. elegans exogenous serotonin induces behavioral changes that are                            observed in the presence of food, whereas exogenous octopamine induces                            behaviors of starved animals [C.                                    elegans [Petrascheck et al. found through chemical                            screening that mianserin, an antidepressant, extends life span of          . They de         . It is s animals . It has  elegans . They fo elegans .       C. elegans through                            mechanisms that are related to food deprivation. We have recently elucidated a                            mechanism for activation of octopamine signaling in the absence of food in C.                                    elegans and demonstrated the involvement of the amine neurotransmitter                            dopamine in this regulation [These results suggest that                            octopamine along with serotonin regulates lifespan in gulation ,9. We reC. elegans,                            activation of CREB can be detected using a cre::gfp fusion gene, in                            which the cyclic AMP response element (CRE) is fused to the gene encoding green                            fluorescent protein (GFP) [cre::gfp, GFP is                            expressed in cells in which CREB is activated. Using this reporter system, we                            first found that the absence of food induces CREB activation in the cholinergic                            SIA neurons [tbh-1 gene were tested. tbh-1 encodes                            tyramine-ƒΐ-hydroxylase which is required for octopamine synthesis and is                            expressed only in the RIC neurons and the gonadal sheath cells (the latter are                            unlikely to play a role in this food response) [tbh-1 mutants                            failed to respond to the absence of food, indicating that octopamine is                            responsible for CREB activation. We also found that exogenous application of                            octopamine in the presence of food induces CREB activation in the SIA neurons,                            which also supports the involvement of octopamine. These results confirmed the                            notion that octopamine signaling is activated in the absence of food.                            Furthermore, we found that the octopamine receptor SER-3 is required for both                            responses to the absence of food and to exogenous octopamine. Cell-specific                            expression of SER-3 in the SIA neurons rescued exogenous octopamine and food responses                            of ser-3 mutant animals, indicating that SER-3 works in the SIA neurons                            to receive octopamine signaling. Given that there is no synaptic connection                            between the RIC and SIA neurons, these results suggest that octopamine released                            from the RIC neurons humorally activates SER-3 in the SIA neurons in the                            absence of food.                                        In in (GFP) . In a st neurons . To deteesponse) . tbh-1 mC. elegans                            is important for food sensing [C. elegans have sensory endings under the cuticle and sense                            the presence of food by mechanosensation [Dopamine signaling in  sensing . Dopaminensation -14. The ensation . Consideensation .       cat-2 mutants, which are                            defective in dopamine synthesis since cat-2 encodes the tyrosine                            hydroxylase, the rate limiting enzyme for dopamine synthesis [cat-2                            mutants exhibited spontaneous CREB activation in the SIA neurons even in the                            presence of food. This spontaneous activation requires endogenous octopamine                            since spontaneous CREB activation was suppressed in cat-2;tbh-1 double                            mutants. These results indicate that octopamine-SER-3-CREB signaling pathway is                            constitutively activated in cat-2 mutants and dopamine normally                            suppresses this pathway in the presence of food.                                        We first found that                            exogenously applied dopamine suppresses exogenous octopamine-mediated CREB                            activation in the SIA neurons. To determine whether endogenous dopamine also                            suppresses octopamine signaling, we tested ynthesis . cat-2 To further demonstrate the                            involvement of endogenous dopamine in the suppression of octopamine signaling,                            we used the Sephadex beads. It was shown previously that the Sephadex beads                            induce a dopamine-dependent behavioral change presumably by mimicking the                            tactile attribute of food without providing nutritional or chemosensory cues                            associated with bacteria (food) . AdditioC. elegans and found that two D2-like                            dopamine receptors, DOP-2 and DOP-3 [We have tested all                            identified dopamine receptors in nd DOP-3 ,17, worknd DOP-3 . Cell-spC. elegans uses a three-neuron-type circuit to control octopamine                            signaling in response to food (Figure These studies suggests thatd Figure . In the C. elegans and a                            three-neuron-type circuit possibly involved in food response in the mammalian                            brain. First, food stimuli increase dopamine and decrease noradrenaline release                            in the mammalian brain [C. elegans.                            Therefore, we postulate that the neuronal and molecular circuitry for food                            sensing we have discovered in C. elegans is conserved in vertebrates, in                            which cessation of dopamine signaling activates octopamine/noradrenaline                            signaling.                                        There are striking                            analogies between this three-neuron-type circuit in an brain ,19. Secoan brain  and theian brain . Third, an brain . This maIt is unknown whether the                            SIA neurons play any role in aging and it is highly possible that                            mianserin-mediated lifespan extension work through its effect on SER-3 in other                            cells. However, the finding that dopamine regulates the octopaminergic RIC                            neurons suggests that octopamine signaling in the cells other than the SIA                            neurons are also regulated by dopamine. This raises the possibility that                            dopamine plays a role in the food-mediated regulation of lifespan.                                        It has been suggested that                            food limits lifespan through at least two different mechanisms. One is by                            providing nutrition and the other is by providing sensory perception .       cat-2 mutants have a normal lifespan when measured in                            standard culture conditions [C. elegans would be                            particularly enlightening, especially since it has been reported that a polymorphism                            in the tyrosine hydroxylase gene, which is required for dopamine synthesis, is                            associated with variation in human longevity [Murakami et al. showed that                            dopamine-deficient nditions . In factnditions ,26. Intrnditions , suggestongevity ,28.    This work was supported in                        part by the Canadian Institutes of Health Research grant MOP-77722 and                        MOP-82909 to J.G.C. J.G.C. and H.H.M.V.T. are holders of Canadian Research                        Chairs. S.S. is a recipient of a Parkinson Society Canada Basic Research                        Fellowship.


Kaposi sarcoma is a low-grade mesenchymal tumor involving blood and lymphatic vessels. There are four variants of this disease, each presenting a different clinical manifestation: classic or sporadic, African or endemic, organ transplant-related or iatrogenic, and AIDS-related or epidemic. Kaposi sarcoma is the most common tumor among patients with HIV infection, occurring predominantly in homosexual or bisexual men. The pulmonary involvement in Kaposi sarcoma occurs commonly in critically immunosupressed patients who commonly have had preceding mucocutaneous or digestive involvement.The etiology of Kaposi sarcoma is not precisely established; genetic, hormonal, and immune factors, as well as infectious agents, have all been implicated. There is evidence from epidemiologic, serologic, and molecular studies that Kaposi sarcoma is associated with human herpes virus type 8 infection. The disease starts as a reactive polyclonal angioproliferative response towards this virus, in which polyclonal cells change to form oligoclonal cell populations that expand and undergo malignant transformation.The diagnosis of pulmonary involvement in Kaposi sarcoma usually can be made by a combination of clinical, radiographic, and laboratory findings, together with the results of bronchoscopy and transbronchial biopsy. Chest high-resolution computed tomography scans commonly reveal peribronchovascular and interlobular septal thickening, bilateral and symmetric ill-defined nodules in a peribronchovascular distribution, fissural nodularity, mediastinal adenopathies, and pleural effusions. Correlation between the high-resolution computed tomography findings and the pathology revealed by histopathological analysis demonstrate that the areas of central peribronchovascular infiltration represent tumor growth involving the bronchovascular bundles, with nodules corresponding to proliferations of neoplastic cells into the pulmonary parenchyma. The interlobular septal thickening may represent edema or tumor infiltration, and areas of ground-glass attenuation correspond to edema and the filling of air spaces with blood. These findings are a result of the propensity of Kaposi sarcoma to grow in the peribronchial and perivascular axial interstitial spaces, often as continuous sheets of tumor tissue.In conclusion, radiological findings can play a major role in the diagnosis of pulmonary Kaposi sarcoma since characteristic patterns may be observed. The presence of these patterns in patients with AIDS is highly suggestive of Kaposi sarcoma. Kaposi sarcoma (KS) was first described by Moritz von Kaposi in 1872 as a low-grade mesenchymal tumor involving blood and lymphatic vessels. The mucocutaneous sites are primarily affected, typically the skin of the lower extremities, face, trunk, genitalia, and oropharyngeal mucosa; other organs are involved in the disseminated form of the disease . This diKS is the most common tumor among patients with human immunodeficiency virus (HIV) infection, occurring predominantly in homosexual or bisexual men ,7. Also,KS is one of the major complications of AIDS . In induCritical immunosupression in patients with mucocutaneous KS commonly leads to pulmonary involvement. Thoracic disease is found in about 45% of patients with cutaneous AIDS-related KS, and in about 15% of patients without mucocutaneous involvement . studiedThe etiology of KS is not precisely established; genetic, hormonal, and immune factors, as well as infectious agents, have all been implicated. There is evidence from epidemiologic, serologic, and molecular studies that KS is associated with human herpes virus type 8 (HHV8) infection ,6,13. InThe presence of KS associated with HHV-8 and host immunosuppression are considered the major factors that promote tumor development -18. The The histopathologic process of the disease is believed to start in the sub epithelial connective tissue, extending in the direction of the epithelium. A developed lesion consists of interwoven bands of spindle cells and vascular structures grouped in a network of reticular and collagen fibers. Erythrocytes are seen within these vascular structures and interspersed between spindle cells. The vascular component appears as small capillaries or slit-like spaces between the spindle cells .Generally, patients with lung KS have previously treated cutaneous lesions or other visceral involvement, for example in the gastrointestinal tract. Nonetheless, cutaneous involvement is absent in 5–23% of patients with symptomatic pulmonary KS . The mosFour clinical variants of KS have been described ,5. The hThe extent of tracheobronchial KS ranges from isolated tracheal lesions to diffuse and/or extensive tracheobronchial involvement. Endobronchial lesions may narrow and partially obstruct the airways. The bronchoscopic appearance of endobronchial KS is considered to be characteristic enough to allow a diagnosis. When bronchoscopic examination identifies KS lesions below the level of the carina, accompanying parenchymal abnormalities are seen, as a rule, on chest radiography or CT. Thus, the appearance of characteristic tracheobronchial KS lesions is sufficient to make a presumptive diagnosis of pulmonary KS ,26.There is increasing evidence that HAART and an improved immune response are associated with complete or partial regression of KS lesions, a decrease in the number of patients suffering from KS, improved survival, and protection of HIV-infected patients against the development of KS ,14. In pThe diagnosis of pulmonary involvement in KS usually can be made by a combination of clinical, radiographic, and laboratory findings, together with the results of bronchoscopy and transbronchial biopsy . KS may Chest radiography may demonstrate middle to lower lung zone reticular opacities and parenchymal nodules with a bronchovascular distribution that may progress to consolidation, peribronchial cuffing, Kerley B lines, pleural collections, and hilar or mediastinal adenopathies  Figure . HRCT scThe sensitivity of HRCT in the diagnosis of endobronchial KS is not high, but tumors large enough to cause stridor in the upper airways or atelectasis in small segmental or lobar bronchi are generally seen as filling defects in the endobronchial lumen. Endoluminal lesions are infrequently detected by HRCT; thickening of the bronchial walls and interstitium, representing tumor infiltration along the interstitium, is much more common ,33.The imaging findings of pulmonary KS reflect the propensity of KS to grow in the peribronchial and perivascular axial interstitial spaces extending to the peripheral regions, often as continuous sheets of tumor tissue Figure . Thus ceSilva Filho et al.  reportedIn conclusion, tomographic findings can play an important role in the diagnosis of pulmonary KS, since characteristic patterns may be observed. The most frequent chest CT findings are thickening of the peribronchovascular interstitium, particularly in the axial portions; irregular and poorly defined nodules; and interlobular septal thickening. The presence of these findings in patients with AIDS is highly suggestive of KS. Correct interpretation of the HRCT patterns, understanding of the histopathological appearance, and association of the histopathological and radiological findings can be very useful to the radiologist in the correct diagnosis of pulmonary KS.KS: Kaposi's sarcoma; AIDS: acquired immunodeficiency syndrome; HIV: human immunodeficiency virus; HHV8: human herpes virus type 8; HAART: highly active antiretroviral therapies.The authors declare that they have no competing interests.TDG conceived the study. SLN, ADV and AASDS performed the literature review. TDG, GZ and EM edit and coordinated the manuscript. All authors read and approved the final manuscript.


EHP issues occasional press releases to publicize papers that we believe are relevant and important to researchers, educators, public health practitioners, policy makers, and/or the general public, as well as to regular EHP readers and contributors. Specifically, our goal is for EHP press releases to serve science and the public by increasing awareness of work that might otherwise be overlooked by people who would benefit from reading it. Of course, press releases may also benefit authors and journals by increasing recognition and citation, which may in turn increase authors’ ability to obtain funding and advance their careers, and increase the journal’s readership and ability to attract high-quality submissions (EHP’s standards for press releases; we would like to take this opportunity to clarify our standards for our readers and contributors.As a service to our contributors and the public, missions . Howevermissions . With th“Exaggeration serves many interests, but it does not serve the public’s interest. And in the end, it is self-defeating, because it undermines the credibility of medical science. After a while people may not believe anything we have to say.”Schwartz and Woloshin 2003Press releases provide an important link between scientists and journalists. Results of a 2005 survey of 468 health and medical science journalists indicated that press releases or press conferences were the initial source of ideas for 40–50% of news stories , and a rThe impact of press releases on media coverage and science communication highlights the need to ensure that releases are accurate, complete, and clear, but systematic reviews of press releases issued by academic centers  and mediEHP’s mission. Science communication to a broad audience may be facilitated by translation and reframing, but we also have a responsibility to communicate in a way that will help improve the public’s ability to understand the implications of environmental health research. With this in mind we will continue to strive to write press releases that present EHP papers in a meaningful and accurate way by putting findings into context without inappropriate extrapolation or exaggeration, and by providing key information on current knowledge, research methods, study limitations, and potential conflicts of interest (EHP receive.Communicating the findings of environmental health research is central to interest . This ap


EHP relies on the diligence and integrity of experts to help determine the quality and impact of papers submitted for possible publication. In 2009 EHP received nearly 1,200 papers, and about 480 of those papers were sent by our Associate Editors to two to four anonymous peer reviewers for evaluation. EHP published 292 papers in 12 issues during 2009, and the journal is very grateful for the time and effort of the more than 1,000 reviewers who assisted us last year. A list of those reviewers is available on the journal’s website (http://ehponline.org/article/info:doi/10.1289/ehp.118-a59).Like all peer-reviewed journals, EHP is recognizing it’s top 12 Reviewers of the Year, individuals who reviewed at least six papers during the year and received excellent ratings for the timeliness and quality of their reviews by the Associate Editor who handled the peer-review process. They are John Balbus, David Carpenter, Deborah Cory-Slechta, Amy Herring, William Kelce, Morton Lippmann, Matthew Longnecker, Arnold Schecter, Rémy Slama, David Thomas, Leo Trasande, and Mary Wolff. We congratulate these reviewers and thank the hundreds of others who contributed to the success of EHP in 2009.For the first time, EHP should note that we have revised our Instructions to Authors (ITA). The revised ITA can be found in this issue (p. A83) and are available online (http://www.ehponline.org/static/instructions.action). Authors should note that the word limit for Letters to the Editor has been reduced from 1,000 to 750 words, and we have tried to make it clear that it is not acceptable to introduce new non–peer-reviewed data in Letters to the Editor. In addition, EHP reserves the right not to publish letters deemed by the editors to be overly personal or polemic.Authors planning to submit manuscripts to EHP now routinely evaluates manuscripts for possible plagiarism. The revised ITA also emphasize our policy of full disclosure of all actual or potential financial or nonfinancial competing interests involving people or organizations that might reasonably be perceived as relevant. Authors must evaluate wording in documents such as grants or contracts to determine whether there might be an actual or potential competing interest. Another major change in the ITA is that EHP will actively evaluate submissions for research that might provide knowledge, products, or technologies that could be directly misapplied to pose a threat to public health and safety.The revised ITA provide more detailed information concerning scientific integrity. Authors should be aware that AMA Manual of Style: A Guide for Authors and Editors in preparing manuscripts. Questions concerning changes in the revised ITA should be directed to EHPmanuscripts@niehs.nih.gov.Other revisions in the ITA include minor changes in guidance for preparation of manuscripts, including tables and figures; updated information concerning the online submission process; and a new embargo policy. In addition, we now recommend that authors refer to the


Environmental Health Perspectives (EHP) as a means of reinforcing high-quality articles published in the journal, identifying emerging research themes, and tracking the impact of groundbreaking research (Tilson 2008). In this issue, we are pleased to announce that the EHP Paper of the Year for 2009 is “Decrease in Anogenital Distance among Male Infants with Prenatal Phthalate Exposure” by Shanna H. Swan, Katharina M. Main, Fan Liu, Sara L. Stewart, Robin L. Kruse, Antonia M. Calafat, Catherine S. Mao, J. Bruce Redmon, Christine L. Ternand, Shannon Sullivan, J. Lynn Teague, and the Study for Future Families Research Team , monobenzyl phthalate (MBzP), and monoisobutyl phthalate (MiBP)] were associated with a shorter AGD. Swan subsequently replicated and extended these findings (2008).Toxicologists routinely measure the external genitalia to assess reproductive toxicity in animal studies. One of these measures, anogenital distance (AGD)—a particularly sensitive indicator of masculinization—is shortened in male rodents following prenatal exposure to several phthalates . Swan etThe AGD measure developed by Environmental Health Perspectives. Our concern about this issue is reflected in our Instructions to Authors, which outlines our expectations concerning full disclosure of competing financial and nonfinancial interests and potential penalties that could be imposed if journal policy is not followed.Integrity in science has been a long-time priority for EHP staff participated in the development of the document mentioned in the editorial, and we fully endorse recommendations concerning the need for full disclosure of competing interests. As further indication of our commitment to maintaining a high degree of scientific integrity, EHP has made the decision to designate a staff member to serve as Ethics Coordinator. The Ethics Coordinator is responsible for ensuring appropriate compliance related to conflict of interest declarations required for each paper, assessing alleged conflicts of interest and plagiarism, and assisting the Editor-in-Chief in assessing potential conflicts of interest of reviewers. Science has made and will continue to make important contributions to the progress of human society. We should, however, be aware that continued support for research is dependent on the perceived integrity of the scientific process. EHP remains dedicated to serving as an independent and unbiased venue for the discussion of the impact of the environment on human health.In this issue is an editorial by Jennifer Sass concerning key elements of effective and practical disclosure policies for health science journals.


Human herpesvirus (HHV)-8 associated oncogenesis, a state of immune impairment, a local inflammatory environment, angiogenesis and HIV infection occurring concurrently are important factors for the development of HIV-associated Kaposi sarcoma (KS).Activation of the interleukin (IL)-6 receptor signalling pathway and constitutive signalling of viral G protein-coupled receptor (vGPCR) play an important role in the activation, proliferation and transformation of HHV-8 infected endothelial cells thus contributing to the initiation and progression of KS. HIV-tat protein, HIV-induced immune suppression and a hyperinflammatory state facilitate the oncogenic activity of HHV-8.In this article we reviewed some aspects of HIV-KS pathogenesis and tried to establish, according to the available information in the literature, whether HIV-KS is a monoclonal neoplasm or a benign angioproliferative disorder.From the data of this review it is evident that most of the HIV-KS lesions are oligoclonal in origin. It remains to be demonstrated whether these multiple monoclonal populations of cells are neoplastic, harbouring specific cytogenetic alterations such as mutations, rearrangements and amplifications, or are, as the current evidence shows, the result of HHV-8 induced intracellular signalling pathways that modulate the expression of cellular genes associated with cell cycle regulation, apoptosis, inflammatory response and angiogenesis, and represent a reactive angioproliferative disorder. The clinical course of human immunodeficiency virus (HIV)-associated Kaposi sarcoma (HIV-KS) ranges in severity from a mild, slowly progressing to a rapidly progressing life threatening condition. Invariably, without treatment, the overall prognosis of HIV-KS is poor .The clonal nature of KS is controversial and the exact cell of origin of KS is not clear. The cell of origin may arise from lymphatic or blood endothelium, vascular smooth muscle cells, mesenchymal cells or a combination of these cells . Howeverbcl-2 family. These biological mediators have the capacity to activate several intracellular signaling pathways that may bring about a hyperinflammatory state, angiogenesis and transformation of endothelial cells. These events are central to the initiation and promotion of HIV-KS [HHV-8 induced inflammation, angiogenesis and oncogenesis are critical for the development of HIV-KS . HHV-8 ef HIV-KS .It is unclear whether HIV-KS is a true neoplastic entity arising from a clonal expansion of a single HHV-8 infected cell, or is a benign angioproliferative disorder. The demonstration that HIV-KS lesions are monoclonal will provide further evidence on the probable malignant nature of HIV-KS, since monoclonality is an important feature of cancer.In this article we reviewed the available data in the literature regarding the clonal nature of HIV-KS.It is probable that HIV-KS starts as a polyclonal inflammatory hyperplasia, and a subset of advanced HIV-KS lesional cells undergo malignant transformation, which may explain the aggressive clinical course of such cases .HHV-8 is aetiologically linked to KS, but is not sufficient on its own to transform HHV-8 infected endothelial cells and to initiate the development of KS. Other factors including HIV-tat protein, a state of immune suppression and dysregulation of cytokines, growth factors and adhesion molecule functions cooperate with HHV-8 to drive the initiation and promotion of HIV-KS [HHV-8 gene expression is tightly controlled and occurs in two phases: the latent phase that facilitates viral genome persistence, and the lytic phase which consists of viral replication, host cell destruction and viral release . During The early KS lesion is most probably initiated by HHV-8 induced production of pro-inflammatory cytokines IL-1, IL-6, tumour necrosis factor (TNF)-α, interferon (IFN)-γ and growth factors basic fibroblast growth factor (bFGF) and vascular endothelial cell growth factor (VEGF) that causes activation of endothelial cells followed by angiogenesis -1, LANA-2, vCyclin, viral Fas-associated death domain-like interleukin 1 gamma-converting enzyme inhibitory protein (vFLIP) and kaposin  that play a role in tumour cell survival during the latent phase of HHV-8 infection ,17.Kaposin is another latent-phase gene product. This unique HHV-8 oncoprotein, encoded by open reading frame (ORF)-K12, is expressed in primary effusion lymphoma cell lines, can induce transformation in vitro, and it has been suggested that it may play a role in the development of KS ,18,19.Some HHV-8 lytic genes Table  play a mThrough constitutive activation of intracellular signaling pathways, vGPCR has the capacity to transform endothelial cells by promoting their uncontrolled proliferation and by inhibition of apoptosis ,15,21,22The increased secretion of angiogenic growth factors, chemokines and cytokines induces either enhanced proliferation of vGPCR-transformed endothelial cells in an autocrine manner, or stimulates angioproliferation of neighboring bystander cells by paracrine mechanisms Diagram  12,14-1,22,24-27Stimulation of vGPCR leads to the activation of NF-κB, activator protein (AP)-1 and nuclear factor of activated T cells (NFAT), which induce the endogenous expression of NF-κB and AP-1 dependant cytokines. These cytokines include IL-1, TNF-α and IL-6 as well as growth factors -1α, a transcription factor, by both the p38 and MAPK signalling pathways. These pathways are associated with VEGF production, and are present in higher levels in HHV-8 transformed endothelial cells  inhibits the interferon signalling pathway and is able to induce tumour formation. The expression of p21, a cyclin dependant kinase (CDK) inhibitor, is downregulated by vIRF, which enables the latent virus to evade the immune mediated IFN cell-cycle shutdown, allowing HHV-8 to establish persistence. However, vIRF is not expressed as much in KS as in the cells of Castleman's disease and primary effusion lymphoma, and its role in the development of KS is not clear .It is probable that other viral lytic oncogenes, such as vIL-6, also play a role in HHV-8 mediated cell cycle dysregulation; however, its role in the sarcomagenesis of KS has not yet been defined .Analysis of the inactivation pattern of HUMARA has been used to determine the clonality of KS lesional cells in females -33. ThisWhile some studies using the HUMARA technique showed that the majority of KS cells were monoclonal ,33, otheJudde and co-workers  studied Duprez et al.,  using thThus KS lesions may be regarded as a reactive proliferation of oligoclonal or monoclonal endothelial cells and not as a true malignancy. The fact that without treatment most subjects with advanced HIV-KS and endemic KS die as a consequence of their KS disease, may be explained by the local aggressive nature of the multicentric lesions. KS lesional cells show uncontrolled proliferation and increased survival. When the lesions affect vital organs such as the heart and the lungs, the disease is fatal .However, one cannot exclude the possibility that in advanced stages of HIV-KS disease, in the presence of other as yet unidentified cofactors cooperating with HHV-8 oncogenes, a subset of the benign monoclonal cells evolve into a malignant clone that undergoes expansion .From the available data in the literature, one can conclude that HIV-KS starts as a reactive polyclonal angioproliferative response towards HHV-8, and with time, some of the polyclonal cells evolve to form oligoclonal cell populations that undergo expansion. This process is driven by HHV-8 induced intracellular signalling pathways that modulate the expression of cellular genes associated with cell cycle regulation, apoptosis, inflammatory responses and angiogenesis. Whether these benign, reactive, multiple monoclonal cell populations, under certain circumstances, undergo malignant transformation remains to be elucidated.


Kaposi's sarcoma is a vascular neoplasm mainly affecting the skin of the lower extremities. Although it is the most common neoplasm affecting patients with AIDS, sporadic cases in HIV-negative people have been reported. It is a lesion mainly affecting men and its clinical presentation presents a challenge, as it can resemble other benign or malignant skin lesions.We report a rare case of Kaposi's sarcoma presenting in a 68-year-old Mediterranean woman with no evidence of HIV infection. The patient had a 6-month history of a slowly progressing pigmented lesion on the dorsum of her left hand. The lesion clinically resembled a squamous cell carcinoma. The patient was treated with a wide excision of the lesion and primary reconstruction with a full thickness skin graft. Histopathological and immunohistochemical analysis of the excised lesion revealed the presence of Kaposi's sarcoma. Serologic investigation for HIV was negative but polymerase chain reaction for human herpes virus type 8 infection was positive. Thorough clinical and imaging investigation of the abdomen and chest were both negative for loci of disease.Kaposi's sarcoma, although rare in its sporadic form, should be considered in the differential diagnosis of indeterminate skin lesions, especially those affecting the extremities. Kaposi's sarcoma (KS) is an angioproliferative skin lesion associated with a great number of epidemiologic and pathophysiologic factors. Due to this variability it is classified into four distinct clinico-epidemiological types: classic Mediterranean KS, African-endemic KS, immunosuppressive drug-related KS and epidemic AIDS-related KS. Despite being the most common neoplasm affecting patients with AIDS, its sporadic presentation is rare and can sometimes escape clinical suspicion.In its classic-sporadic type, KS presents as a cutaneous lesion typically affecting the skin of the extremities. Epidemiologically it is most often observed in elderly patients but it is a rare occurrence in females. The incidence is higher in Southern and Eastern European countries and the condition is more commonly found in Jewish, Italian and Greek populations.A 68-year-old woman presented with a 6-month history of a slowly evolving, asymptomatic, raised, slightly pigmented skin lesion, measuring 25–30 mm in diameter, involving the dorsum of her left hand Figure . CutaneoThe surgical specimen was submitted for pathological evaluation. Histological and immunohistological findings were consistent with a diagnosis of KS Figure  and 3B. Following the diagnosis the patient was subjected to a thorough diagnostic evaluation to determine the possible spread of the disease to other sites. Chest X-ray, abdominal ultrasonography, upper and lower gastrointestinal endoscopy, as well as thoracic and abdominal computed tomography, were all negative for the presence of disease.Wide local excision with histologically negative margins is regarded as the accepted method of treating minimal cutaneous lesions of KS. Since no dissemination of the disease was demonstrated in the postoperative clinical and radiological evaluation, no further treatment modalities were considered necessary. During a 9-month follow-up, no local or distant recurrence was observed. Re-evaluation for HIV seroconversion was negative.Since its first description, KS has remained a tumour of undetermined pathogenesis. There is still doubt regarding the nature of the proliferating cells and whether the lesions represent an exuberant hyperplasia or a neoplasm. KS lesions have a number of peculiar characteristics, including a lack of aneuploidy and a strong association with immunodeficiency. Emergence of KS in transplant recipients and immunosuppressed patients is remarkable because it may regress spontaneously if immunosuppression is reduced or discontinued. These characteristics suggest that it is not a malignant neoplasm but a benign, potentially controllable and reversible hyperplasia ,2. On thThe pathogenesis of KS is still under investigation. Current studies have focused on the search for a causative infectious agent mainly due to its correlation with immunocompromised patients. HHV8 infection is widespread in Mediterranean areas with a relatively high incidence of KS . A large body of evidence has strongly linked all KS clinical variants with HHV8 infection .HHV8 DNA is present in all forms of KS but not in other mesenchymal tumours or non-specific inflammatory lesions of the skin, suggesting a strong association of HHV8 and KS . MoreoveThe presence of HHV8 infection is now considered essential for the development of KS. However, the incidence of HHV8 infection is far higher than the prevalence of KS, suggesting that viral infection per se is not adequate for the development of malignancy and that one or more cofactors are necessary. The exact interactions of HHV8 infection with other factors known to be involved in the pathogenesis of KS, including anti-apoptosis genes and cytokines, which may be mediated by virally encoded genes as well as gender, genetic susceptibility, and immunosuppression, still need clarification .A high HHV8 seroprevalence in individuals engaging in high-risk sexual activity, including homosexual men, indicates the role of sexual behaviour in the transmission of the infection in adults . On the The HHV8 reservoir in peripheral blood appears to be mononuclear cells and its detection in mononuclear cells is predictive of KS development . A sensiClinically, the classic form of KS is restricted mainly to the surface of the body. Three distinct stages of progression, which can overlap, can be recognized: patch, plaque, and nodule. In the first stage, a clinically indistinct red-to-blue macule occurs, usually in the lower extremity. KS may be mistaken in the skin for an inflammatory dermatosis, pyogenic granuloma, angiodermatitis or pseudo-Kaposi's, bacillary angiomatosis, angiosarcoma, bullous lesion in the rare cases of lymphangioma-like or bullous KS, or arteriovenous malformations. In the case we have presented here, a raised, slightly pigmented skin lesion was initially regarded as a squamous cell carcinoma. The lesion's clinical presentation, its location, the otherwise normal skin, and the lack of any predisposing factors with the exception of the patient's origin, were misleading.The best treatment modality is still a matter of debate and no standard treatment guidelines are available. Many patients have cutaneous lesions amendable to local therapy . Some patients require more aggressive local therapy (radiation therapy) or systemic therapies . Yet surgical excision has been frequently associated with local recurrence, while chemotherapy with vinblastin or bleomycin has been characterized as not very effective . In contIn our case, a wide local excision was performed because the lesion was regarded as a squamous cell carcinoma, for which standard surgical excision is the preferred treatment. If we had considered the possibility of KS, we would have chosen an incisional biopsy to determine the tumour histology. Excisional biopsy would not have been the preferable method because the lesion was too large to anticipate primary closure.No matter what treatment modality is chosen, clinicians should bear in mind that even in its classical form, KS may be a malignant, rapidly progressing tumour with visceral involvement. Although primary hand KS is a relatively uncommon disorder in patients who are negative for HIV, dermatologists, venereologists, and surgeons should consider this possibility when treating non-specific lesions involving the extremities. Since various effective treatment options are available, watchful waiting is probably inappropriate in most cases.KS is an unusual vascular tumour that, similarly to other forms of cutaneous neoplasms, has a definite evolutionary course from a low-grade slow-growing early phase, to an anaplastic malignant neoplasm. Skin biopsy is important for making the correct diagnosis, with the added use of immunohistochemistry or molecular biology in equivocal cases.Minimal hand lesions with non-distinctive clinical features may be the exclusive manifestation of KS, making clinical suspicion essential and histological evaluation necessary to establish the diagnosis.HHV8: human herpes virus type 8; KS: Kaposi's sarcoma; PCR: polymerase chain reaction.The authors declare that they have no competing interests.Written informed consent was obtained from the patient for publication of this case report and any accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal.All the authors have contributed equally to this case report. All authors read and approved the final manuscript.


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