Acute oral toxicity in mice of a new palytoxin analog: 42-hydroxy-palytoxin.

The acute oral toxicity of a new palytoxin congener, 42-hydroxy-palytoxin (42-OH-PLTX), was investigated in female CD-1 mice. The toxin (300-1697 µg/kg), administered by gavage, induced scratching, jumping, respiratory distress, cyanosis, paralysis and death of mice, with an LD50 of 651 µg/kg (95% confidence limits: 384-1018 µg/kg) within 24 h. Hematoclinical analyses showed increased plasma levels of lactate dehydrogenase and aspartate-aminotransferase at doses of 600 µg/kg and above, as well as of alanine-aminotransferase, creatine phosphokinase and potassium ions at ≥ 848 µg/kg. Histology revealed inflammatory lesions in the non-glandular area of the stomach of mice that survived up to 24 h after gavage (424-1200 µg/kg). Although no histological alterations were seen in skeletal and cardiac muscles, changes in some plasma biomarkers (creatine phosphokinase, lactate dehydrogenase) suggested involvement of these tissues in 42-OH-PLTX oral toxicity, in agreement with epidemiological data on seafood poisonings ascribed to palytoxins. Complete recovery of the tissue and hematological changes was observed two weeks post-exposure. Furthermore, 42-OH-PLTX induced in vitro delayed erythrocyte hemolysis at concentrations similar to those of PLTX (EC50 = 7.6 and 13.2 x 10⁻¹² M, respectively). This hemolysis could be completely neutralized by a monoclonal anti-PLTX antibody. The in vivo data, together with the in vitro data recorded for 42-OH-PLTX, seem to indicate Na+/K+-ATPase as one of the key cellular targets of this toxin.

Detection of 16 gamma-ray pulsars through blind frequency searches using the Fermi LAT.

Pulsars are rapidly rotating, highly magnetized neutron stars emitting radiation across the electromagnetic spectrum. Although there are more than 1800 known radio pulsars, until recently only seven were observed to pulse in gamma rays, and these were all discovered at other wavelengths. The Fermi Large Area Telescope (LAT) makes it possible to pinpoint neutron stars through their gamma-ray pulsations. We report the detection of 16 gamma-ray pulsars in blind frequency searches using the LAT. Most of these pulsars are coincident with previously unidentified gamma-ray sources, and many are associated with supernova remnants. Direct detection of gamma-ray pulsars enables studies of emission mechanisms, population statistics, and the energetics of pulsar wind nebulae and supernova remnants.

A population of gamma-ray millisecond pulsars seen with the Fermi Large Area Telescope.

Pulsars are born with subsecond spin periods and slow by electromagnetic braking for several tens of millions of years, when detectable radiation ceases. A second life can occur for neutron stars in binary systems. They can acquire mass and angular momentum from their companions, to be spun up to millisecond periods and begin radiating again. We searched Fermi Large Area Telescope data for pulsations from all known millisecond pulsars (MSPs) outside of globular clusters, using rotation parameters from radio telescopes. Strong gamma-ray pulsations were detected for eight MSPs. The gamma-ray pulse profiles and spectral properties resemble those of young gamma-ray pulsars. The basic emission mechanism seems to be the same for MSPs and young pulsars, with the emission originating in regions far from the neutron star surface.

A long-period, violently variable X-ray source in a young supernova remnant.

Observations with the Newton X-ray Multimirror Mission satellite show a strong periodic modulation at 6.67 +/- 0.03 hours of the x-ray source at the center of the 2000-year-old supernova remnant RCW 103. No fast pulsations are visible. If genetically tied to the supernova remnant, the source could either be an x-ray binary, composed of a compact object and a low-mass star in an eccentric orbit, or an isolated neutron star. In the latter case, the combination of its age and period would indicate that it is a peculiar magnetar, dramatically slowed down, possibly by a supernova debris disc. Both scenarios require nonstandard assumptions about the formation and evolution of compact objects in supernova explosions.

Phase-resolved spectroscopy of Geminga shows rotating hot spot(s).

Isolated neutron stars are seen in x-rays through their nonthermal and/or surface thermal emissions. X-ray Multimirror Mission-Newton observations of the Geminga pulsar show a 43-electron volt spectrum from the whole neutron star surface, as well as a power-law component above 2 kiloelectron volts. In addition, we have detected a hot (170 electron volts) thermal emission from an approximately 60-meter-radius spot on the pulsar's surface. Such a thermal emission, only visible at selected phase intervals, may be coming from polar hot spot(s), long thought to exist as a result of heating from magnetospheric accelerated particles. It may provide the missing link between the x-ray and gamma-ray emission of the pulsar.

Geminga's tails: a pulsar bow shock probing the interstellar medium.

We report the X-ray Multimirror Mission-Newton European Photon Imaging Camera observation of two elongated parallel x-ray tails trailing the pulsar Geminga. They are aligned with the object's supersonic motion, extend for approximately 2', and have a nonthermal spectrum produced by electron-synchrotron emission in the bow shock between the pulsar wind and the surrounding medium. Electron lifetime against synchrotron cooling matches the source transit time over the x-ray features' length. Such an x-ray detection of a pulsar bow shock (with no Halpha emission) allows us to gauge the pulsar electron injection energy and the shock magnetic field while constraining the angle of Geminga's motion and the local matter density.

The magnetic field of an isolated neutron star from X-ray cyclotron absorption lines.

Isolated neutron stars are highly magnetized, fast-rotating objects that form as an end point of stellar evolution. They are directly observable in X-ray emission, because of their high surface temperatures. Features in their X-ray spectra could in principle reveal the presence of atmospheres, or be used to estimate the strength of their magnetic fields through the cyclotron process, as is done for X-ray binaries. Almost all isolated neutron star spectra observed so far appear as featureless thermal continua. The only exception is 1E1207.4-5209 (refs 7-9), where two deep absorption features have been detected, but with insufficient definition to permit unambiguous interpretation. Here we report a long X-ray observation of the same object in which the star's spectrum shows three distinct features, regularly spaced at 0.7, 1.4 and 2.1 keV, plus a fourth feature of lower significance, at 2.8 keV. These features vary in phase with the star's rotation. The logical interpretation is that they are features from resonant cyclotron absorption, which allows us to calculate a magnetic field strength of 8 x 10(10) G, assuming the absorption arises from electrons.

Pfiesteria: review of the science and identification of research gaps. Report for the National Center for Environmental Health, Centers for Disease Control and Prevention.

In connection with the CDC National Conference on Pfiesteria, a multidisciplinary panel evaluated Pfiesteria-related research. The panel set out what was known and what was not known about adverse effects of the organism on estuarine ecology, fish, and human health; assessed the methods used in Pfiesteria research; and offered suggestions to address data gaps. The panel's expertise covered dinoflagellate ecology; fish pathology and toxicology; laboratory measurement of toxins, epidemiology, and neurology. The panel evaluated peer-reviewed and non-peer-reviewed literature available through June 2000 in a systematic conceptual framework that moved from the source of exposure, through exposure research and dose, to human health effects. Substantial uncertainties remain throughout the conceptual framework the panel used to guide its evaluation. Firm evidence demonstrates that Pfiesteria is toxic to fish, but the specific toxin has not been isolated or characterized. Laboratory and field evidence indicate that the organism has a complex life cycle. The consequences of human exposure to Pfiesteria toxin and the magnitude of the human health problem remain obscure. The patchwork of approaches used in clinical evaluation and surrogate measures of exposure to the toxin are major limitations of this work. To protect public health, the panel suggests that priority be given research that will provide better insight into the effects of Pfiesteria on human health. Key gaps include the identity and mechanism of action of the toxin(s), the incomplete description of effects of exposure in invertebrates, fish, and humans, and the nature and extent of exposures that place people at risk.

[The impact on the international literature of the scientific production of Italian researchers in the disciplines "psychiatry" and "psychology". A bibliometric evaluation]. / L'impatto sulla letteratura internazionale della produzione scientifica dei ricercatori italiani, nelle discipline "psichiatria" e "psicologia". Una valutazione bibliometrica.

OBJECTIVE: The aim of the study was to present the results of a citation analysis concerned with the impact of Italian researchers and institutions in psychiatry and psychology upon the international scientific community. METHOD: The analysis has been performed using a database of the Institute for Scientific Information (ISI): All scientific papers which were published between 1981 and 1998 in psychiatric and psychological journals included in the Science Citation Index (SCI) and the Social Sciences Citation Index (SSCI) were considered. The most cited Italian papers, authors and institutions are reported, as well the most frequently utilised journals. RESULTS: Publications concerned with neuropsychology, psychopharmacology and biological psychiatry were the most cited. This prevalence also affected the ranking of the most cited authors, even though, in this case, research groups in disciplines such as clinical psychology and epidemiological psychiatry appeared to be strong. The four most productive Italian Universities were characterized by the presence of both a School of Medicine and a School of Psychology. The Journal of Neurology, Neurosurgery and Psychiatry and Psychopharmacology were the most frequent vehicles of scientific communication. CONCLUSIONS: The results entail important implications for Italian research in psychology and psychiatry. On a general level, these analyses appear to be helpful for monitoring scientific production by granting agencies and for comparing different individual researchers. On a more specific level the analysis has identified the leading trends in research of Italian psychiatry and psychology.

Evidence for palytoxin as one of the sheep erythrocyte lytic in lytic factors in crude extracts of ciguateric and non-ciguateric reef fish tissue.

The occurrence of palytoxin or its congener in fish extracts has been presented in this study. The presences of hemolytic factors in fish extracts of Hawaiian reef fish and their implication in ciguatera poisoning have been shown by the sheep erythrocyte assay. By use of the anti-palytoxin inhibition assay with fish extracts and sheep red blood cell (RBC), it was shown that palytoxin was one of the major factors in the lysis of sheep erythrocytes. Ouabain, an antagonist of palytoxin for the Na+/K+ ATPase receptor on RBC, also showed inhibition of sheep RBC lysis by fish extracts. From these results, it was concluded that, in part, palytoxin and other palytoxin-related, hemolysin-like factors in fish extracts were responsible for sheep cell hemolysis.

[The physician-patient relationship in the contexts of different medicines]. / Il rapporto medico-paziente nei contesti delle diverse medicine.

The quality of the doctor-patient relationship depends at least in part on the way in which various non-specific factors influence disease and healing processes, which applies in particular to the subjects' beliefs (attributions) concerning what causes or prevents disease and the efficacy of various possible remedies. This favours the alternative medicines since in the exchanges with the patients, their models of etiopathogenesis and of mode and mechanism of action of proposed remedies come much closer to common sense models than those of modern scientific medicine. Such an advantage is increased by the fact that at least 80% of the encounters between physicians and patients concern situations of malaise or discomfort not identifiable as specific diseases. In such situations, official medicine often fails to exert the necessary functions of listening, explaining, counselling and reassuring, but tends to an inappropriate use of diagnostic and therapeutic tools developed for specific pathologies. Therefore, the dialogue between the different medicines, by promoting the re-establishment of a patient-centered approach, can increase the efficacy of the interventions which depends both on specific technical factors and the quality of the relationship.

[Ethics problems in phase IV of drug studies]. / Problemi etici nella fase IV degli studi sui farmaci.

Phase IV clinical studies include all investigations carried out after the approval of drugs. The objective of these studies is well defined: to gain additional knowledge on efficacy and safety of drugs. There are uncertainties however with regard to which kind of study design is appropriate to provide scientifically valid contributions. In the present article we will clarify why an experimental design (e.g. randomized clinical trial) is ordinarily required, on the basis of scientific and ethical concerns, to answer questions concerning efficacy even after drug commercialization. Viceversa, non experimental designs (from case series analysis to cohort and case control studies) are indicated to investigate drug effects in current practice if the objective is to evaluate safety.

Monoclonal antibodies to taxanes that neutralize the biological activity of paclitaxel.

Recently it has been proposed that drug-specific neutralizing antibodies may limit side-effects that occur during chemotherapy. These studies were undertaken to determine if monoclonal antibodies, 3C6 specific to paclitaxel and 8A10 specific to taxane diterpenes, are inhibitors of paclitaxel-induced inhibition of proliferation and cellular microtubule and nuclear changes. The results show that 3C6 and 8A10 each inhibit paclitaxel-induced cytotoxicity, microtubular bundling, stabilization from vinblastine-induced microtubule depolymerization and the formation of micronuclei. We conclude that these antibodies effectively neutralize paclitaxel activity in vitro and that they may be useful to determine if antibody-blocking strategies can prevent dose-limiting toxicities.

Antibodies to maitotoxin elicited by immunization with toxin-fragment conjugates.

Maitotoxin is a water-soluble polyether toxin produced by the benthic dinoflagellate, Gambierdiscus toxicus. Toxin fragments generated by periodate oxidation were conjugated to keyhole limpet hemocyanin, and the resulting immunogens were used to elicit maitotoxin-specific antibodies in mice. A competitive immunoassay developed with polyclonal IgG antibodies detected approximately 3 nM purified maitotoxin standard (IC50 approximately 13 nM; 45 ng/ml) but did not detect other polyether marine toxins. These are the first reported maitotoxin-specific antibodies.

Biological activity of 26-succinylbryostatin 1.

Bryostatin 1, a macrocyclic lactone, has undergone phase I trials as an anticancer agent. Because of the lipid solubility of this compound it must be delivered either in ethanol or in a PET formulation. During the trial, these vehicles caused a large number of treatment-related side effects. We have synthesized the triethanolamine salt of 26-succinylbryostatin 1 and find that this compound is approx. 100-fold more water soluble than bryostatin 1. Because of the potential for clinical use, we have evaluated the biologic activity of this compound. We find that in a concentration-dependent manner 26-succinylbryostatin 1 is capable of activating protein kinase C (PKC) in vitro and displacing [3H]PDBu from PKC. However, at all concentrations tested the activity was less than the parent compound bryostatin 1. Addition of bryostatin 1 but not 26-succinylbryostatin 1 to U937 leukemic cells in culture stimulated a drop in cytosolic PKC, secondary to translocation of PKC to the membrane. Although 26-succinylbryostatin 1 did not stimulate a drop in the cytosolic levels of PKC, addition to U937 cells activated transcription from an AP-1 enhancer construct and c-Jun protein phosphorylation in a similar fashion to bryostatin 1 and differentiation of U937 cells. Unlike bryostatin 1, 26-succinylbryostatin 1 was unable to cause aggregation of human platelets. Although injection of bryostatin-1 into mice carrying B16 melanoma inhibits tumor growth, there was no significant inhibition of melanoma growth when identical doses of 26-succinylbryostatin 1 were injected. Therefore, 26-succinylbryostatin 1 shares some but not all of the pharmacologic properities of bryostatin 1. This compound can activate protein phosphorylation without lowering cytosolic levels of PKC.

Economical test methods for developmental neurobehavioral toxicity.

The assessment of behavioral changes produced by prenatal or early postnatal exposure to potentially noxious agents requires both the designing of ad hoc tests and the adaptation of tests for adult animals to the characteristics of successive developmental stages. The experience in designing tests is still more limited than in the adaptation of tests, but several tests have already proven their usefulness; some examples are the suckling test, the homing test, and evaluations of dam-pup and pup-pup interactions. Functional observational batteries can exploit the development at specified postnatal ages of several reflexes and responses that are absent at birth in altricial rodent species with a short pregnancy such as the rat and the mouse. In neonates, the assessment of early treatment effects can rely not only on deviations from normal responding but also on changes in the time of appearance of otherwise normal response patterns. The same applies to other end points such as responses to pain and various types of spontaneous motor/exploratory activities, including reactivity to a variety of drug challenges that can provide information on the regulatory systems whose development may be affected by early treatments. In particular, the analysis of ontogenetic dissociations (i.e., differential early treatment effects depending jointly on developmental stage at the time of exposure, age of testing, and response end point) can be of considerable value in the study of treatments' mechanisms of action. Overall, it appears that behavioral teratological assessments can be effectively used both proactively, i.e., in risk assessment prior to any human exposure, and reactively. In the latter case, these assessments could have special value in the face of agents suspected to produce borderline changes in developing humans, whose innocuousness or noxiousness can be difficult to establish in the absence of hard evidence of teratogenicity.

Activation of stress-activator protein kinase/c-Jun N-terminal kinase by the non-TPA-type tumor promoter palytoxin.

Previous studies have shown that structurally diverse tumor promoters can modulate protein kinases involved in signal transduction. In this study, we show that palytoxin, a potent non-12-O-tetradecanoylphorbol-13-acetate (TPA)-type skin tumor promoter, induces a signaling pathway leading to the activation of the stress-activated protein kinases/c-Jun N-terminal kinases (JNK) in Swiss 3T3 fibroblasts. Treatment of cells with doses as low as 0.1 mN palytoxin results in significant activation of JNK. In contrast to epidermal growth factor, which induces a transient activation of JNK in Swiss 3T3 cells, palytoxin causes prolonged enzyme activation. Since stimulation of ion flux appears to play an important role in the mechanism of action of palytoxin in other systems, we investigated the role of sodium and calcium in the activation of JNK: (a) our results show that incubation of Swiss 3T3 cells in a sodium-free medium dramatically reduced the magnitude of JNK activation by palytoxin; and (b) we found that the sodium ionophore gramicidin activates JNK. Together, these results suggest that sodium influx, which is a hallmark of palytoxin action, may play a key role in the activation of JNK by palytoxin. Our results indicate that calcium influx is not necessary or sufficient for palytoxin-induced activation of JNK. In contrast to palytoxin, the TPA-type tumor promoter phorbol 12,13-dibutyrate and the non-TPA-type tumor promoters thapsigargin and okadaic acid do not appear to activate JNK in this system. In contrast to phorbol 12,13-dibutyrate, palytoxin does not activate the p42/p44 mitogen-activated protein kinases. Our results demonstrate that Swiss 3T3 fibroblasts, palytoxin can activate a protein kinase signaling pathway that is distinct from that activated by the prototypical phorbol ester tumor promoters and other potent skin tumor promoters.